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DR ANTHONY MELVIN CRASTO Ph.D

DR ANTHONY MELVIN CRASTO, Born in Mumbai in 1964 and graduated from Mumbai University, Completed his Ph.D from ICT, 1991,Matunga, Mumbai, India, in Organic Chemistry, The thesis topic was Synthesis of Novel Pyrethroid Analogues, Currently he is working with GLENMARK PHARMACEUTICALS LTD, Research Centre as Principal Scientist, Process Research (bulk actives) at Mahape, Navi Mumbai, India. Total Industry exp 30 plus yrs, Prior to joining Glenmark, he has worked with major multinationals like Hoechst Marion Roussel, now Sanofi, Searle India Ltd, now RPG lifesciences, etc. He has worked with notable scientists like Dr K Nagarajan, Dr Ralph Stapel, Prof S Seshadri, Dr T.V. Radhakrishnan and Dr B. K. Kulkarni, etc, He did custom synthesis for major multinationals in his career like BASF, Novartis, Sanofi, etc., He has worked in Discovery, Natural products, Bulk drugs, Generics, Intermediates, Fine chemicals, Neutraceuticals, GMP, Scaleups, etc, he is now helping millions, has 9 million plus hits on Google on all Organic chemistry websites. His friends call him Open superstar worlddrugtracker. His New Drug Approvals, Green Chemistry International, All about drugs, Eurekamoments, Organic spectroscopy international, etc in organic chemistry are some most read blogs He has hands on experience in initiation and developing novel routes for drug molecules and implementation them on commercial scale over a 30 year tenure till date Dec 2017, Around 35 plus products in his career. He has good knowledge of IPM, GMP, Regulatory aspects, he has several International patents published worldwide . He has good proficiency in Technology transfer, Spectroscopy, Stereochemistry, Synthesis, Polymorphism etc., He suffered a paralytic stroke/ Acute Transverse mylitis in Dec 2007 and is 90 %Paralysed, He is bound to a wheelchair, this seems to have injected feul in him to help chemists all around the world, he is more active than before and is pushing boundaries, He has 9 million plus hits on Google, 2.5 lakh plus connections on all networking sites, 50 Lakh plus views on dozen plus blogs, He makes himself available to all, contact him on +91 9323115463, email amcrasto@gmail.com, Twitter, @amcrasto , He lives and will die for his family, 90% paralysis cannot kill his soul., Notably he has 19 lakh plus views on New Drug Approvals Blog in 216 countries......https://newdrugapprovals.wordpress.com/ , He appreciates the help he gets from one and all, Friends, Family, Glenmark, Readers, Wellwishers, Doctors, Drug authorities, His Contacts, Physiotherapist, etc

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CIFORADENANT


img

Structure of CIFORADENANT

CIFORADENANT

1202402-40-1
Chemical Formula: C20H21N7O3
Molecular Weight: 407.434

CPI-444, CPI 444, CPI444, V81444, V-81444, V 81444,

UNII 8KFO2187CP

 Corvus Pharmaceuticals, Inc. PHASE 1

(S)-7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine

3H-1,2,3-TRIAZOLO(4,5-D)PYRIMIDIN-5-AMINE, 7-(5-METHYL-2-FURANYL)-3-((6-((((3S)-TETRAHYDRO-3-FURANYL)OXY)METHYL)-2-PYRIDINYL)METHYL)-

(73 S)-15 -methyl-6-oxa-2(7,3)-[1,2,3]triazolo[4,5- d]pyrimidina-4(2,6)-pyridina-1(2)-furana-7(3)- oxolanaheptaphan-25 -amine adenosine receptor antagonist

Ciforadenant, also known as CPI-444 and V81444, is an orally administered antagonist of the adenosine A2A receptor. Upon oral administration, CPI-444 binds to adenosine A2A receptors expressed on the surface of immune cells, including T-lymphocytes, natural killer (NK) cells, macrophages and dendritic cells (DCs). This prevents tumor-released adenosine from interacting with the A2A receptors on these key immune surveillance cells, thereby abrogating adenosine-induced immunosuppression in the tumor microenvironment.str1

Ciforadenant is an antagonist of adenosine A2A being developed by Corvus , under license from Vernalis , for the oral treatment of advanced solid tumor; the company is also developing the drug in combination with atezolizumab , for non-small-cell lung cancer.

In 2015, Vernalis licensed the exclusive rights of the product for use of all therapeutic application to Corvus.

Synthesis

WO 2009156737

PATENT

WO 2009156737

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=F7135D4AE9D62AF12284DD6C449A0E96.wapp1nC?docId=WO2009156737&tab=PCTDESCRIPTION&queryString=EN_ALL%3Anmr+AND+PA%3Avernalis+&recNum=42&maxRec=288

US 8450328

WO2017112917

WO 2018175473

WO 2018009972

WO 2018049271

WO 2018022992

PATENT

WO 2018013951

PATENT

WO-2018183965

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2018183965&redirectedID=true

EXAMPLES

Reaction Scheme 1

[0314] Referring to Reaction Scheme 1 , the process to manufacture triazolo[4,5]pyramidine derivatives and intermediates thereof in accordance with the present disclosure, such as the compound known as CPI-444, consists of three chemical steps and uses starting materials known as CP-55, CP-56 and CP-60. The intermediate known as CP-57 is formed at step la without isolation (telescoped) and taken to the next step to form the compound known as CP-58 at step lb. Suzuki coupling using CP-60 during step 2 generates crude CPI-444 which undergoes crystallization during step 3 to form CPI-444.

[0315] Previously described processes for making triazolo[4,5]pyramidine derivatives and intermediates thereof utilized a compound known as CP-59:

[0316] Moreover, such previously described process utilize triethylamine which takes a longer time for the layers to separate where excessive rag layer is observed during phase separation. [0317] The present inventors unexpectedly and surpisingly found that the replacement of CP-59 with CP-60 improved ease of handling and improved process efficiency. In addition, the present inventors unexpectedly and surpisingly found that the use of potassium carbonate (K2CO3) during step 2 improves the phase separation and minimizes rag layer formation upon reaction completion. Finally, Step 3 employs the use of thermocycler in order to facilitate the removal of residual solvents such as isopropyl alcohol.

[0318] Accordingly, the processes in accordance with the teachings of the present disclosure are an improvement over, and are more suitable for commercial scale-up, than processes previously described.

[0319] Starting material (C-55) is commercially available through Astatech, Inc., Keystone Business Park, 2525 Pearl Buck Road, Bristol, PA, 19007, USA; or Suven, SDE Serene Chambers, Road No.5, Avenue 7 Banjara Hills, Hyderabad, 500034, India.

[0320] CP-60 is commercially available through ARK Pharma, Inc., 3860 North Ventura Drive, Arlington Heights, IL, 60004, USA; or Boron Technology Institute, Road No. 2, Building No. 10, room No. 259, Haidian District, Beijing, China.

EXAMPLE 1. Preparation of CP-56

Reaction Scheme 1


Boc20, CbzCI

[0321] Preparation of Dimethyl pyridine-2,6-dicarboxylate:

Pyridine-2,6-dicarboxylic acid (900g, leq) is suspended in methanol(5 volume) and added H2SO4. (19g). The mixture is heated to reflux for approximately 4hr. After reaction completion, the mixture is cooled to 5- 10°C to allow the solids to precipitate. The solids are stirred for an additional hour. The solids are collected by filtration. The wet-cake is re-dissolved in DCM (3 volume) and extract in sequence with an aqueous saturated solution of NaHC03 (2 Volume) followed by with a 5% brine solution (2 Volume). The organic layer is concentrated to dryness to obtain dimethyl pyridine-2,6-dicarboxylate; 914.85g, purity 100%, yield 87.%.

[0322] Preparation of pyridine-2,6-diyldimethanol:

Dimethyl pyridine-2,6-dicarboxylate (885g, leq) is dissolved in EtOH (4425g, 5 Volume) at room temperature. The NaBH4 (341 g, 2eq) is added slowly to the reaction while keeping the internal temperature below 30°C using an ice bath. The reaction is heated to 35°C for approximately 2hrs. After reaction completion, the mixture is cooled to room temperature and adjusted with 32% HCl solution to pH value of approximately 2.5. The mixture is stirred for

2hrs to allow the solids to precipitate. The mixture is then adjusted pH value of approximately 9 using 30% NaOH solution while maintaining an internal temperature below 30°C and stirred at room temperature for about 30 min. The solids are removed by filtration. The filtrate is concentrated at 50°C. The concentrated residual is suspended with isopropanol (4160g, 8 vol)

/water (416g, 0.8 vol) and heated to 70°C for about lhr. The solution is then cooled to room

temperature and stirred for 2hr before cooling to 5-10°C for 30min. The un-dissolved solids are

removed by filtration. The filtrate is concentrated at 50°C. The concentrated residue is charged

with dichloromefhane (2700g, 5vol) and heated to 40 °C for 30min. The suspension is cooled to 5-

10°C and stirred for 30mins. The solid is collected by filtration and dried under vacuum at 40°C to obtain pyridine-2,6-diyldimethanol; 540.77g, purity 100%, yield 85.86%.

[0323] Preparation of 2,6-6 s(chloromethyl)pyridine:

2,6-bis(chloromethyl)pyridine (400g, leq) is suspended in DCM (2000g) and then cooled to 10- 15°C. Thionyl chloride (SOCb; 775g, 3eq) is charged with CH2CI2 (775g) and then added drop- wised into the reaction vessel while maintaining the internal temperature below 20 °C. The reaction is then warmed to room temperature and held for approximately 2hrs. After reaction completion, the 15% aqueous solution of a2C03 (9038g) is pre-cooled to 10-15°C before charging the reaction mixture into the carbonate solution while maintaining internal temperature below 20 °C. The mixture is stirred until gas-evolution is no longer observed. The organic layer is extracted with water (2 x 3200g) and then concentrated at 50°C to a crude product. The concentrated crude is purified by recrystallization using heptane (946g). The mixture is cooled to 5-10°C for 30min. The solid is collected by filtration and wet-cake is washed with heptane and dried at 40°C under vacuum to obtain 2,6-6zs(chloromethyl)pyridine; 442.6g, purity 100%, yield 87.0%.

[0324] Preparation of (3r,5r,7r)-l-((6-(chloromethyl)pyridin-2-yl)methyl)-l,3,5,7-tetraazaadamantan-l-ium:

2,6-to(chloromethyl)pyridine (420g, leq) is dissolved in CH2CI2 (8400g), HMTA (336g, leq) is added into the reaction vessel. The reaction is heated to approximately 40 °C for about 3hrs. Additional HMTA (168g, 0.5eq) is added into the reaction mixture and stirred overnight at room

temperature. The product is collected by filtration. The wet-cake is washed with CthCkand dried under vacuumat 50°C to obtain (3r,5r,7r)-l -((6-(chloromethyl)pyridin-2-yl)methyl)- 1 ,3, 5,7-tetraazaadamantan- 1 -ium; 730g, purity 97.01%, yield 96.58%.

[0325] Preparation of (6-(chloromethyl)pyridin-2-yl)methanamine dihydrochloride:

(3r,5r,7r)- 1 -((6-(chloromethyl)pyridin-2-yl)methyl)- 1 ,3 ,5 ,7-tetraazaadamantan- 1 -ium (730g, leq) is suspended in EtOH (4380g) before charging 37% HC1 (159g). The mixture is heated to approximately 60 °C for about lhr. After reaction completion, it is cooled to 25°C. MTBE

(1200g) is charged into the suspension. The suspension is then stirred for about 30 min and cooled to 5-10°C for about lhr. The solids are collected by filtration and washed with MTBE and dried at 50°C under vacuum to obtain (6-(chloromethyl)pyridin-2-yl)methanamine dihydrochloride; 449.56g (after assay correction), purity 98.15%, yield85.23%.

[0326] Preparation of tert-butyl ((6-(chloromethyl)pyridin-2-yl)methyl)carbamate:

(6-(chloromethyl)pyridin-2-yl)methanamine dihydrochloride [422.56g (after assay correction), leq] is dissolved in CH2CI2 (5600g) and pre-cooled to 10-15°C. K2CO3 (1632g) pre-dissolved in water (4000g) is charged into the reaction solution solution. The mixture is stirred for about lOmin and then cooled to 10-15°C. Boc-anhydride (603g) is pre-dissolved in CH2CI2 (1808g) before charging into the reactor. The mixture is warmed to room temperature and held for about an hour. After reaction completion, the organic layer is extracted with water (4000g), The organic layer is concentrated to dryness at 50 °C to obtain tert-butyl ((6-(chloromethyl)pyridin-2-yl)methyl)carbamate; 382.93g [after assay correction); purity 99.01%; yield 81%].

[0327] Preparation of tert-butyl ((6-(iodomethyl)pyridin-2-yl)methyl)carbamate:

tert-butyl ((6-(chloromethyl)pyridin-2-yl)methyl)carbamat [ 382.93g (after assay correction) , leq] is dissolved in THF (1 150) and Nal (720g) is added, the reaction is at room temperature for approximately 4hr. After reaction completion, excess Nal and NaCl are filtered off and the filtrate is concentrated at 40°C. The concentrated residue is re-dissolved in ethyl acetate (2300g) and extracted with water (2900g), the organic layer is washed with 10% aqueous solution of Na2S203 (2600g) followed by 5% brine solution (2900g). The organic layer is concentrated to a residue. The residue is re-dissolved in ethyl acetate (4200g), and then filtered. The filtrate is oncentrated and taken up in ethyl acetate (765g) and stirred at room temperature for about 2hr before slowly adding heptane (380g). The solids are filtered and dried at 50°C under vacuum to

obtain tert-butyl ((6-(iodomethyl)pyridin-2-yl)methyl)carbamate; 440g; purity 100%, Yield 85%.

[0328] Preparation of tert-butyl (S)-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)carbamate:

A solution of t-BuOK (113g in THF (1.1 kg) is pre-cooled to 5- 10°C, before charging asolutionof (S)-tetrahydrofuran-3-ol (166g) in THF (220g). The mixture is stirred at room temperature for about lhr. A solution of tert-butyl ((6-(iodomethyl)pyridin-2-yl)methyl)carbamate (440g, leq) in THF (880g) is pre-cooled to 10-15°C before. The tetrahydrofuranyl solution is slowly charged into reaction solution while maintaining an internal temperature below 1 °C. After about 1 hour another solution of pre-cooled solution of t-BuOK (50g) and (S)-tetrahydrofuran-3-ol (66g) in THF (405g) kg) is slowly added into reaction mixture while maintaining internal temperature below 10 °C. The mixture is stirred at about 10 °C for approximately 1 hour. After reaction completion, the mixture is quenched with water (2200g) and extracted with toluene (4400g). The organic layer is washed with 5% brine (2x 2200g). The organic layer is concentrated to dryness at 50°C under vacuum to obtain tert-butyl (S)-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)carbamate; 389g, purity 89.63%, yield 105%.

[0329] Preparation of CP-56 free base:

tert-butyl (S)-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)carbamate (389g, leq) is dissolved in CH2CI2 (1556g) and pre-cooled to 0-5°C before charging drop-wise methanesulfonic acid ( MSA; 600g) into the reaction solution while maintaining internal temperature below 20°C. The mixture is warmed to room temperature and hold for about lhr. After reaction completion, water (389g) is added and cooled to 5-10°C. 30% NaOH is charged to adjust the reactor pH to approximately 12.5. The mixture is stirred for about 30 min before extracting with CH2CI2 (1556g). The organic layer is collected and extracted with an aqueous saturated solution of brine (584g). The organic layer is concentrated under vacuum. The residue is re-dissolved in toluene (1560g andthenconcentrated. The concentrated residue is re-dissolved in toluene (1560g) and then filtered. The filtrate is concentrated to dryness at 50°C under vacuum to obtain CP-56 free base; 221g (after assay correction), purity 91%, yield 84.23%.

[0330] Preparation of CP-56:

CP-56 free base (22 lg (after assay correction), leq) is dissolved in MeOH (260g) and EtOH (1300g) and then cooled about 15°C. Oxalic acid (47), pre-dissolved in MeOH (1 lOg is charged into reaction mixture. The reaction is at 15-20°C for 3hr. The mixture is cooled to 0-5°C and

stirred for about an Ihr. The solid is collected by filtration and the wet-cake is washed with EtOH (390g). The solid is dried under vacuum at 50°C to obtain CP-56 crude. Crude CP-56 is re-crystallized from isopropanol (865g) and H20 (lOOg). The mixture is heated to about 70°C to obtain a solution. The solution is slowly cooled to 50°C for Ihr. The mixture is cooled to 0-5°C for about another Ihr. The solid is filtered and washed with isopropanol. The wet-cake is dried at 50°C under vacuum to obtain CP-56; 164g, purity 99%, yield 95%.

[0331] Alternatively, CP-56 can be formed using the following process:

Reaction Scheme 2

7 8 9

[0332] Preparation of Dimethyl pyridine-2,6-dicarboxylate (compound 2):

Charge diacid (1; 628g) into reactor containing methanol (2Kg) and heat to reflux. After reaction completion the reaction is cooled to 30 C and stirred. The wet-cake is filtered and washed with methanol (500g). The wet-cake is dried under vacuum at about 55 °C to obtain diester (680 g, purity >99%; yield 85%).

[0333] Preparation of 6-(hydroxymethyl)picolinamide (compound 4):

Charge diester (2; 600 g) into reactor containing methanol (1.8 kg) and tetrahydrofuran (1.2 kg). Charge slowly sodium borohydride ( aBH4; about 130 g) into the reaction solution while maintaining an internal temperature below 30 °C. After reaction completion aqueous hydrochloric acid (about 350 g of 32% HC1) is charged into the reaction solution. The mixture is concentrated and then charged with dichloromethane (1.8 kg). The organic solution is extracted with water (600 g) and then concentrated to obtain the crude product (3). Crude 3 was dissolved in methanol (1.3 kg) and then charge ammonium hydroxide (20%; 1.3 kg). The solution was stirred until reaction completion before concentrating solution. The residue was taken up in water (600g) and heated to about 60 °C before cooling to 0 °C. The wet-cake was filtered, washed with water and dried in vacuum oven to obtain 6-(hydroxymethyl)picolinamide (about 220 g, >99% purity).

[0334] Preparation of 6-(chloromethyl)picolinonitrile (compound 5):

Charge 6-(hydroxymethyl)picolinamide (about 220 g) into a rector containing acetonitrile (450 g). Charge POCb (519 g and agitate at about 70 °C. After reaction completion the solution is

cooled to about 30 °C before slowly charging into a pre-cool (about 10 °C) reactor with water

(305 g). Charge toluene (1.4 kg) to extract the solution mixture. The toluene phase is washed in sequence with 20 % NaOH (600 g), saturated NaHC03 (300 g) and water (300 g). Toluene is concentrated to obtain crude Cl-nitrile, 5. Isopropyl alcohol (400 g) is charged to dissolve the wet-cake at about 45 °C before cooling to about 0 °C. The wet-cake was filtrated and washed with heptane (150 g) and dried in vacuum oven to obtain 6-(chloromethyl)picolinonitrile (180 g; > 99%.

[0335] Preparation of (S)-6-(((tetrahydrofuran-3-yl)oxy)methyl)picolinonitrile (compound 7):

Charge Cl-nitrile (180 g) into a rector containing THF (540 g). Charge Nal (185.7 g) to the reactor and stirred at 50 °C. After reaction completion, the reactor is cooled to 0 °C. In another

reactor, charge t-BuOK (145.6 g) and THF (320 g). Add (S)-tetrahydrofuran-3-ol (31 1.9 g) into the reactor while maintaining internal temperature below 50 °Cto deprotonate the alcohol. Stir

until t-BuOK dissolves. Add THF-OK / THF solution into 6-(iodomethyl)picolinonitrile solution (compound 6) while maintaining internal temperature below 10 °C. Stir at room

temperature until reaction completion. Concentrate the solution to remove THF solvent. Add

ethyl acetate (630 g) and wash by water (420 g). Extract water phase by ethyl acetate (630 g). Combine organic layer and concentrate to obtain oil crude 374 g. The residue was distilled under vaccum (P=3~4 torr, internal temperature 174 °C to 188 °C) to obtain (S)-6-

(((tetrahydrofuran-3-yl)oxy)methyl)picolinonitrile (compound 7) as an oily product (204g, >96% purity; 74% yield).

[0336] Preparation of (S)-(6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methanamine (compound 9):

Charge (S)-6-(((tetrahydrofuran-3-yl)oxy)methyl)picolinonitrile (180 g) into a rector containing MeOH (1620 g). Charge NaOMe (95.3 g) to the reactor and stirred for 30 min at 30 °C until

reaction completion. The methyl (S)-6-(((tetrahydrofuran-3-yl)oxy)methyl)picolinimidate solution (compound 8) was transferred to hydrogenation apparatus containing 50% Ni (60 g). Purge with N2 and then increase the H2 pressure. Under H2 pressure of 5 kg / cm2 and temperature of 30 °C until reaction completion. The reaction is filtered through celite. The filtrate is concentrated. Toluene is charged (1kg) and then concentrated. Then add toluene (1000 g) and filter to remove salt by-products. The filtrate was concentrated to obtain the oil residue of (S)-(6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methanamine (136 g; 85% yield, assay 80%, >91% purity).

[0337] Preparation of CP-56:

Charge (S)-(6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methanamine (170 g) into a rector containing isopropyl alcohol (600 g). Set internal temperature of 75 °C. In another reactor,

charge oxalic acid (41.1 g) and water (60 g) and heat solution. Add oxalic acid solution into

CP-56 free-base solution. Cool to 30 °C for about 4 hours and agitate. The wet-cake was filtered

and washed with isopropyl alcohol (175 g) and dried under vacuum drying with heat to obtain crude CP-56 (136.2 g). Charge CP-56 crude (123 g) into a rector containing methanol (1295 g). Stir until CP-56 was dissolved completely. Filter through celite to remove insoluble salt. The filtrate is concentrated. Charge isopropyl alcohol (500 g) and water (50 g) to dissolve CP-56 using heat. Cool to about 30 °C for about 3 hours and stir. The wet-cake was filtrated and

washed by isopropyl alcohol (165 g) and dried under vacuum drying with heat to obtain CP-56 (1 13.4 g. purity = >99 %, > 99% ee).

EXAMPLE 4. Preparation of CPI-444

CP-58 CP-60

C15H16CIN702 CPI-444

1H-17BO3

W: 361 .79 MW: 208.06 C20H21N O3

MW: 407.43

[0349] It is to be noted that other Pd coupling reagents can also be used such as Pd(PPh3)4 or Pd(PPh3)2Cl2.

[0350] A solution of CP-58 (30.0 g, 1 equiv.), CP-60 (approximately 20.8 g, 1.2 equiv.), in THF (approximately 180 mL), K2C03 (approximately 17.5 g), Pd(dtbpf)Cl2(approximately 337 mg), and water (approximately 100 mL) were stirred and heated to about 60 °C until reaction completion. The reaction was cooled to about 50 °C and the layers were allowed to separate. The aqueous layer was removed and back extracted with THF (approximately 30 mL). The THF layers were combined and water (approximately 450 ml) was added to precipitate out crude CPI-444. The slurry was cooled to about 20 °C and stirred for approximately 60 min and the slurry was filtered. The cake was washed in sequence with water (approximately 120 ml) and 2-propanol (approximately 30 ml). The wet-cake was dried in the vacuum oven to provide an off- white solid (29.74 g, 88% yield) with a purity of 98.5 %. Crude CPI-444 conforms to reference.

-444 can be prepared by the following process:

EDA and DAP are used to remove Palladium during CPI-444 formation.

[0352] The solution of CP-58 (10 g), CP-60 (6.9 g) , Pd(dtbpf)C12 (approx. 0.0015 mol eq) and K2C03 (5.8 g) in THF (6V) and H20 (3V) is heated to approximately 60 °C. The reaction is complete after approximately 30 minutes. The solution is cooled to 50 °C and aqueous layer is separated. The aqueous layer is extracted with THF (9 mL); the THF layer is added to organic solution. The organics are cooled to 40 °C, 1 ,3-diaminopropane (DAP; approximately 50 g) or ethylene diamine (EDA; approximately 45 g) is added and the mixture stirred for 1 hour. H20 (15V) is added to the organic layer over 10 min. The slurry is cooled to 20 °C for 2 hours, and stirred for an additional 1 hour. The slurry is filtered and washed with H20 (2V x 2) and zPrOH (IV). CPI-444 wet-cake is dried at 50 °C under full vacuum. (Yield = 90 %; purity > 99.0%).

[0353] Alternatively, CPI-444 can be prepared by the following process:

using cysteine in TNF to remove Palladium during CPI-444 formation

[0354] CP-58 (1 kg), K2C03 (0.58 kg), water (3 kg), CP-60 (0.69 kg), and THF (5.3 kg),

Pd(dtbpf)Cb (3 g). The solution is heated to 60 °C. The reaction is complete after approximately 30 minutes. Charge THF (4.5 kg) and cool to 50 °C. The aqueous layer is separated. The organic layer is charged with cysteine (0.32 kg) and water (5 kg). The mixture is agitated. NH4OH (1.1 kg) is charged to the reaction mixture and agitate for approximately 15 minutes. The layers are allowed to separate and the lower aqueous layer is separated. The organic layer is charged with cysteine (0.32 kg) and water (5 kg). The mixture is agitated. NH4OH (1.1 kg) is charged to the reaction mixture and agitate for approximately 15 minutes. The layers are allowed to separate and the lower aqueous layer is separated. THF is distilled to approximately 7 volumes under atmospheric pressure. The solution is cooled to 50 °C before charging NH4OH (0.5 kg) and agitate for 30 min. Water (14.5 kg) is charged while maintaining the internal temperature >40 °C. The reactor is cooled to 20 °C for 2 hours and hold for an additional 1 hour. CPI-444 is filtered and washed with water followed by isopropanol. CPI-444 wet-cake is dried under vacuum at 50 °C. Purity > 99%, yield 85%.

EXAMPLE 5. Removal of Residual Palladium With Biocap Filter Cartridge

[0355] A mixture of CPI-444 crude (16.00 g), THF (approximately 190 ml), L-cysteine

(approximately 8 g), and H20 (approximately 90 ml) were mixed and heated to a solution at about 60 °C for 1 hour. A solution of 28% NH OH (approximately 20 ml) was added and heated for an additional 15 minutes. The agitation was turned off to allow the layers allowed to settle. The aqueous layer was removed; the THF layer was washed with brine solution (approximately 15 ml). The combined aqueous solutions were back extracted with THF (approximately 15 ml). A 3M Biocap filter (BC0025LR55SP; available from 3M) was pretreated with THF (approximately 150 ml) at about 50 °C. The combined organic layers were recirculated through the Biocap at about 10 ml/min for approximately 3 hours and then filtered forward. The Biocap filter was rinsed with THF (approximately 130 ml) at about 50 °C. The combined filtrates were concentrated. Water

(approximately 80 ml) was added, and distilled to remove residual THF. 2-Propanol (approximately 1 10 ml) was added to the slurry, and the mixture was heated to a solution. The solution was cooled to 20 °C and water (approximately 240 ml) was added. The slurry was performed in series by heating to about 55 °C and held that that temperature for approximately 30 minutes, cooled to 20 °C over 30 minutes, and held at 20 °C for 30 minutes. This heating cycle was repeated two more. The slurry was then held at 20 °C for approximately 12 hours. The slurry was filtered, and the product was washed with water (approximately 300 ml). The wet cake (about 23 g) was dried in the vacuum oven to obtain an off white solid (13.6 g; 85% yield;99.9% purity; Pd = 25 ppm).

[0356] Reprocess of step 4. AFC-825-106

[0357] CPI-444 (16.02 g, AFC-825-48) and THF (approximately 280 ml) were charged to a flask and heated to about 50 °C for about 30 minutes to obtain a solution. A 3M Biocap filter

(BC0025LR55SP) was pretreated with THF (approximately 150 ml) at about 50 °C . The CPI-444 solution was passed through the Biocap at aboutl O ml/min. The Biocap filter was rinsed with THF (approximately 130 ml) at about 50 °C. The combined filtrates were transferred to a reactor and concentrated. Water (approximately 80 ml) was added, and distilled to remove residual THF solvent. 2-Propanol (approximately 1 10 ml) was added to the slurry and heated to about 65 °C to obtain a solution. The solution was cooled to about 20 °C before adding water (approximately 240 ml). The slurry was heated to 55 °C over 30 minutes, held at 55 °C for 30 minutes, cooled to 20 °C over 30 minutes, and held at 20 °C for 30 minutes. This heating cycle was two more times. The slurry was then held at 20 °C for 12 hours. The slurry was filtered, and the product was washed with water (approximately 300 ml). The wet cake (26.6 g) was dried in the vacuum oven overnight to obtain 15 as a white solid (95% yield; 99% purity; Pd = 5 ppm).

EXAMPLE 6. Removal of Residual Palladium With Darco KB-G

Crude CPI-444

CPI-444 Drug Substance

[0358] Crude CPI-444 (475 g, 1.17 mol, 1.00 eq), 2-MeTHF (1 1.9 L, 25.0 vol) and WFI water (2.6 L, 5.5 vol) were charged to a 19 L jacketed reactor. The mixture was mechanically agitated under a nitrogen blanket. Nitrogen was bubbled through the solution for 20 minutes. L-Cysteine (242 g, 1.99 mol, 1.71 eq) was then charged. The solution in the reactor was heated to 55±5 °C. Upon reaching 50 °C, the reaction mixture was stirred for 1 hour. 28-30% NH4OH (594 mL, 1.25 vol) was charged via addition funnel, and then the reaction mixture was stirred for 15 min. Agitation was stopped and the reaction was allowed to separate for 1 hour. The aqueous layer was removed. The organic layer was allowed to cool to ambient. The organic layer was filtered and the frit was washed with 2-MeTHF (618 mL, 1.3 vol). The organics were concentrated off by rotary evaporation. WFI water (2.42 L, 5.1 vol) and IPA (2.38 L, 5.0 vol) were used to charge the concentrated slurry to a clean 19 L jacketed reactor under N2. The mixture was heated to 65±5 °C, and then was stirred for 1 hour to obtain solution. Darco KB-G activated carbon (71.3 g, 15 wt%) was charged. The reactor was heated to 75±5 °C and stirred for 15 hours. A I L pocket filter was prepared with filter cloth and a heating jacket and heated to 70±5 °C. Reactor contents were filtered through the pocket filter using N2 pressure. The pocket filter was rinsed with a mixture of IPA/WFI water (1 : 1, 950 mL, 2 vol) followed by a mixture of IPA/WFI water (1 : 1, 1.90 L, 4 vol) and IPA/WFI water ( 1 : 1 , 1.90 L, 4 vol). Inside a 22 L three neck round bottom flask the filtrates were mechanically agitated under a N2 blanket. WFI water (7.13 L, 15 vol) was slowly added via addition funnel over 1 h at ambient temperature, and aged for 1 h. The slurry was heated to 55±5 °C and maintained the temperature for 30 min. This heating and subsequent cooling were repeated twice more. After reaching ambient

temperature the final time, the mixture was stirred for at least 2 hours. The reaction mixture was filtered and the reactor rinsed with WFI water (2.38 L, 5.0 vol, 3x). The cake was dried under N2 for 30 minutes and then transferred to a glass dish. The material was dried under full vacuum at 55±5 °C. The desired product was obtained 368.1 g (77%) as light yellow solids. This material was 99.6% pure by HPLC and had a Pd content of 3.6 ppm.

EXAMPLE 7. Removal of Residual Palladium With Polymer-Bound Thiol (SiST)

[0359] Crude CPI-444 (24.48 g, pd = 1267 ppm) and THF (244.8 mL, 10 vol) were charged to a 500 mL 4-necked flask fitted with mechanical agitation, a condenser with nitrogen balloon and a thermometer. The slurry was heated to 60 °C for 20 minutes and then slowly cooled to 45 °C. SiST (36.72 g) was added to the solution and the mixture was stirred at 42 °C for 14 h. The mixture was filtered and washed by THF (24 mL, 1 vol, twice; Pd= 13.12 ppm). H20 (120 mL, 5 vol) and IPA (120 mL, 5vol) were charged to the flask. The slurry was heated to 70 °C and maintained for 1 h (the slurry became solution). The solution was slowly cooled to room temperature and the slurry was added H20 (360 mL, 15 vol) and heated to 55 °C for 1 h. The slurry was cooled to room temperature and then heated to 55 °C for 1 h. The slurry was cooled to rt. and stirred at rt. for 2 h. The slurry was filtered and washed by H20 (100 mL, 4 vol, three times). The wet cake (28.36 g) was dried by 10 mmHg and 50 °C for overnight (14h) and the weight of CPI-444 was 19.31 g (79% recovery).

EXAMPLE 8. Removal of Residual Palladium By Recrystallization

[0360] CUNO Filter Cartridge 55 S

[0361] CPI-444 (5.0 g, Pd 14.06 ppm) and THF (50 mL, 10 vol) were charged to a 100 mL 3-necked flask fitted with stirring bar, a condenser with nitrogen balloon and a thermometer. The slurry was heated to 60 °C for 20 minutes and added CUNO 55S filter (0.75 g, 15w%). The mixture was stirred at 60 °C for 1 h. The mixture was filtered and washed by THF (5 mL, 1 vol, twice). The filtrate was concentrated. The solid, H20 (25 mL, 5 vol) and IPA (25 mL, 5vol) were charged to 250 mL 3 -necked flask fitted with stirring bar, a condenser with nitrogen balloon and a thermometer. The slurry was heated to 70 °C and maintained for 1 h (the slurry became solution). The solution was slowly cooled to rt.(40 minutes) The slurry was added H20 (75 mL, 15 vol) and then heated to 55 °C for 1 h. The slurry was cooled to rt. (30 minutes) and stirred at rt. for 2 h. The slurry was filtered and washed by H20 (20 mL, 4 vol, three times). The cake (6.355 g) was dried by 10 mmHg and 50 °C

for overnight (16 h) and the weight of CPI-444 was 4.281 g (85% recovery). Pd content(ppm) = 2.02 ppm.

[0362] Polymer-bound Thiol: SiST

[0363] CPI-444(5 g; Pd 14.06ppm) was dissolved in THF (50 mL) at 60 °C. The solution was cooled to 55 °C and SiST (7.5 g) was added to the solution. The solution was stirred at 50-55 °C for 16 h. The solution was filtered through celite and a 0.2 micron filter. The filtrate was tested for Pd content. Result: 2.43 ppm.

Catalyst

Molecular Weight: 291.6990

Molecular Weight: 337.3430

[0364] 1. A solution of S.M., CP-60, Pd(PPh3)2Cl2 and K2C03 in THF – H20 (7.9 mL, 1 : 1) was put in oil-bath at 70-75 °C.

[0365] 2. After 2 h, 0.047 g CP-60 was added to the reaction at 70-75 °C.

[0366] 3. After 1 hr, the reaction was cooled to rt. and 10 mL H20 was added to the reaction.

[0367] 4. The reaction was filtered to provide wet cake (0.812 g).

[0368] 5. The solid wet cake was dried at 45 °C and 20 mmHg for 2h to provide weight 0.499 g. (86%).

[0369] 6. The solid wet cake was stirred in 2 mL DMF for 30 mins (slurry) and then filtered. The solid was dried by 45 °C and 10 mmHg for 12h to provide weight 0.40 g; 69% yield; 98.1% purity.

//////////CIFORADENANT, CPI-444, CPI 444, CPI444, V81444, V-81444, V 81444, UNII 8KFO2187CP,  Corvus Pharmaceuticals, Inc.,  PHASE 1, 

NC1=NC2=C(N=NN2CC3=NC(CO[C@H]4CCOC4)=CC=C3)C(C5=CC=C(O5)C)=N1

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Molidustat, Bay 85-3934


Molidustat structure.png

Molidustat

UNII-9JH486CZ13, cas no 1154028-82-6, MW: 314.3076

2-(6-morpholin-4-ylpyrimidin-4-yl)-4-(triazol-1-yl)-1H-pyrazol-3-one

Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitors

  • Originator Bayer Schering Pharma
  • Developer Bayer HealthCare Pharmaceuticals
  • Class Antianaemics; Morpholines; Pyrazoles; Pyrazolones; Pyrimidines; Small molecules; Triazoles
  • Mechanism of Action Hypoxia-inducible factor-proline dioxygenase inhibitors
  • Phase III Anaemia
  • 24 Jun 2018 Biomarkers information updated
  • 23 Jun 2018 Bayer initiates enrolment in the MIYABI HD-M phase III trial for Anaemia in Japan (PO) (NCT03543657)
  • 05 Jun 2018 Bayer plans a phase III trial for Anaemia (renal) in Japan in June 2018 (NCT03543657)

For the cardio-renal syndrome, a Phase IIb program with the investigational new drug Molidustat (BAY 85-3934) is under initiation in patients with anemia associated with chronic kidney disease and/or end-stage renal disease. Molidustat is a novel inhibitor of hypoxia-inducible factor (HIF) prolyl hydroxylase (PH) which stimulates erythropoietin (EPO) production and the formation of red blood cells. Phase I data have shown that inhibition of HIF-PH by Molidustat results in an increase in endogenous production of EPO.

About Bayer HealthCare

The Bayer Group is a global enterprise with core competencies in the fields of health care, agriculture and high-tech materials. Bayer HealthCare, a subgroup of Bayer AG with annual sales of EUR 18.6 billion (2012), is one of the world’s leading, innovative companies in the healthcare and medical products industry and is based in Leverkusen, Germany. The company combines the global activities of the Animal Health, Consumer Care, Medical Care and Pharmaceuticals divisions. Bayer HealthCare’s aim is to discover, develop, manufacture and market products that will improve human and animal health worldwide. Bayer HealthCare has a global workforce of 54,900 employees (Dec 31, 2012) and is represented in more than 100 countries. More information at www.healthcare.bayer.com.

molidustat

Molidusat sodium

2D chemical structure of 1375799-59-9

RN: 1375799-59-9
UNII: CI0NE7C96T

Molecular Formula, C13-H13-N8-O2.Na, Molecular Weight, 336.2897

Sodium 1-[6-(morpholin-4-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1H-pyrazol-5-olate

Molidustat sodium is an orally-available hypoxia-inducible factor prolyl hydroxylase inhibitor in phase I clinical trials at Bayer for the treatment of patients suffering from renal anemia due to chronic kidney disease.

Molidustat (INNBay 85-3934) is a drug which acts as a HIF prolyl-hydroxylase inhibitor and thereby increases endogenous production of erythropoietin, which stimulates production of hemoglobin and red blood cells. It is in Phase III clinical trials for the treatment of anemia secondary to chronic kidney disease.[1][2] Due to its potential applications in athletic doping, it has also been incorporated into screens for performance-enhancing drugs.[3]

WO 2008067871

WO 2012065967

WO 2013167552

2-Heteroaryl-4-aryl-1,2-dihydropyrazolones having a bactericidal and/or fungicidal action are disclosed in EP 165 448 and EP 212 281. The use of 2-heteroaryl-4-aryl-1,2-dihydropyrazolones as lipoxygenase inhibitors for treatment of respiratory tract, cardiovascular and inflammatory diseases is claimed in EP 183 159. 2,4-Diphenyl-1,2-dihydropyrazolones having a herbicidal activity are described in DE 2 651 008.

The preparation and pharmacological properties of certain 2-pyridyl-1,2-dihydropyrazolones are reported in Helv. Chim. Acta 49 (1), 272-280 (1966). WO 96/12706, WO 00/51989 and WO 03/074550 claim compounds having a dihydropyrazolone partial structure for treatment of various diseases, and hydroxy- or alkoxy-substituted bipyrazoles for treatment of neuropsychiatric diseases are disclosed in WO 2006/101903.

Heteroaryl-substituted pyrazole derivatives for treatment of pain and various CNS diseases are furthermore described in WO 03/051833 and WO 2004/089303. WO 2006/114213 has meanwhile disclosed 2,4-dipyridyl-1,2-dihydropyrazolones as inhibitors of HIF prolyl 4-hydroxylases.

The x-ray crystal structure of the compound 3-methyl-1-(pyridin-2-yl)-4-(1-pyridin-2-yl-3-methyl-1H-pyrazol-5-yl)-2H-3-pyrazolin-5 (114)-one (other name: 5,5′-dimethyl-2,2′-di-pyridin-2-yl-1′,2′-dihydro-2H,3′H-3,4′-bipyrazol-3′-one) is reported inActa Crystallogr., Section E: Structure Reports Oμline E57 (11), o1126-o1127 (2001) [Chem. Abstr. 2001:796190].

The synthesis of certain 3′,5-dimethyl-2-phenyl-1′-(1,3-thiazol-2-yl)-1′H,2H-3,4′-bipyrazol-5′-ol derivatives is described inIndian J. Heterocyclic Chem. 3 (1), 5-8 (1993) [Chem. Abstr. 1994:323362].

The preparation and tautomerism of individual 4-(pyrazol-5-yl)-pyrazolin-5-one derivatives is reported in J. Heterocyclic Chem. 27 (4), 865-870 (1990) [Chem. Abstr. 1991:428557]. A therapeutic use has not hitherto been described for the compounds mentioned in these publications. The compound 2-tert-butyl-1′-[4-(4-chlorophenyl)-1,3-thiazol-2-yl]-3′,5-dimethyl-1′H,2H-3,4′-bipyrazol-5′-ol is listed as a test example in WO 2007/008541.

SYN

WO 2013167552

CLIP

https://onlinelibrary.wiley.com/doi/pdf/10.1002/cmdc.201700783

Image result for molidustat

1-[6-(Morpholin-4-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1Hpyrazol-5-ol (molidustat, BAY 85-3934, 45): Method A (gram-scale): Ethyl 3-(dimethylamino)-2-(1H-1,2,3-triazol-1-yl)acrylate (73, 1.98 g, 9.43 mmol) and 4-(6-hydrazinopyrimidin-4-yl)morpholine (78, 1.89 g, 9.70 mmol) were introduced into ethyl acetate (25 mL) and TFA (502 mg, 4.4 mmol) was added at RT. The mixture was stirred under reflux for 18 h, then cooled to 0–58C and subsequently stirred for a further 2 h. The solid formed was filtered off, washed with cold ethyl acetate and dried first in air and thereafter under a high vacuum. Yield: 2.13 g (71%);

1H NMR (400 MHz, [D6 ]DMSO): d=8.42 (s, 1H), 8.38 (s, 1H), 8.01 (s, 1H), 7.73 (s, 1H), 7.70 (s, 1H), 3.71–3.65 (m, 4H), 3.57–3.51 ppm (m, 4H);

13C NMR (125 MHz, [D6 ]DMSO): d=44.3, 65.6, 85.6, 102.8, 123.7, 132.9, 135.8, 152.4, 154.1, 154.7, 162.0 ppm;

IR (KBr): n˜ =3441, 3135–3108, 2965–2884, 1636–1345, 1257 cm@1 ;

UV/Vis (acetonitrile/water 1:1): lmax (e)= 249 nm (34928 L (mol cm)@1 );

MS (EI+) m/z: 315 [M+H]+ ;

Anal. calcd for C13H14N8O2 : C 49.7, H 4.5, N 35.7, O 10.2, found: C 49.5, H 4.4, N 35.5, O 12.6.

Method B (kilogram-scale): Inastirred vessel, 4- (6-hydrazinopyrimidin-4-yl)morpholine (78, 42.0 kg, 215.1 mol) and methyl 3-(dimethylamino)-2-(1H-1,2,3-triazol-1-yl)acrylate (83, 44.0 kg, 224.2 mol) were suspended in ethyl acetate (378 kg), admixed with TFA (12.1 kg, 106.1 mol) and heated under reflux (from 788C to 81 8C) at a jacket temperature of 908C for 26 h. The suspension obtained was cooled to 0 8C, stirred at 08C for 1 h and filtered. The filter cake was washed with ethyl acetate (53 kg) and dried under reduced pressure at up to 458C. The filter cake was admixed with a mixture of water (355 kg) and acetic acid (11.7 kg), then suspended and stirred at 50–548C for 1 h. After cooling to 248C, the suspension was filtered. The filter cake was washed first with water (90 kg), then twice with methanol (50 kg each time) and finally dried at 35–458C under reduced pressure. Yield: 57.4 kg (85%)

Synthesis of molidustat sodium (84)

Sodium 1-[6-(morpholin-4-yl)pyrimidin-4-yl]-4-(1 H-1,2,3-triazol1-yl)-1H-pyrazol-5-olate (molidustat sodium, 84): Kilogram scale: In a stirred vessel, compound 45 (55 kg, 175.0 mol) was suspended in a mixture of methanol (200 kg) and water (30 kg), admixed with triethylamine (17.8 kg, 175.9 mmol), heated at 608C, stirred further for about 1 h and filtered hot to separate off undissolved constituents. The filter cake was washed with methanol (15 kg, 608C). Sodium hydroxide solution (18.7 kg, 210.4 mmol, 45% strength) was slowly introduced at 608C and methanol (5 kg) was added. Sodium 1-[6-(morpholin-4-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)- 1H-pyrazol-5-olate (84, 0.12 kg) was added as seed crystals and the mixture was stirred at 608C for another 1 h and cooled to 248C over a period of about 2 h. The mixture was stirred for 8 h at this temperature, subsequently cooled to 08C over a period of about 1 h and filtered in portions by means of a centrifuge. The filter cake was washed with a mixture of water (24 kg) and methanol (168 kg) and also methanol (about 23 kg in each case) and dried all together at 40 8C under reduced pressure in a dryer for 8 h. Yield: 57.6 kg (98%);

1H NMR (500 MHz, [D6 ]DMSO): d=8.98 (d, J= 1.4 Hz, 1H), 8.72 (s, 1H), 8.68 (s, 1H), 8.64 (d, J=1.4 Hz, 1H), 7.77 (s, 1H), 4.25–4.00 ppm (m, 8H);

13C NMR (125 MHz, [D6 ]DMSO): d= 48.2, 67.8, 91.5, 107.0, 129.6, 130.9, 138.0, 151.7, 152.0, 157.4, 159.9 ppm;

IR (KBr): n˜ =3153–3006, 2976–2855, 1630–1439, 1241, 1112, 987 cm@1 ;

UV/Vis (acetonitrile/water 1:1): lmax (e)=284 nm (16855 L [mol cm]@1 );

MS (EI+) m/z: 337 [M+Na]+ , 315 [M+H]+ ;

Anal. calcd for C13H13N8O2Na: C 46.4, H 3.9, N 33.3, found: C 46.1, H 4.0, N 33.1.

PATENT

RM 1

Example 3A 3-(Dimethylamino)-2-(1H-1,2,3-triazol-1-yl)acrylic acid ethyl ester

Figure US20100305085A1-20101202-C00024

The preparation of the starting compound is carried out analogously to 2A starting from 1.00 g (6.45 mmol) 2-(1H-1,2,3-triazol-1-yl)acetic acid ethyl ester.

Yield: 1.4 g (100% of th.)

1H-NMR (400 MHz, DMSO-d6): δ=8.10 (d, 1H), 7.78 (d, 1H), 7.65 (s, 1H), 4.03 (q, 2H), 3.06 (br. s, 3H), 2.10 (br. s, 3H), 1.12 (t, 3H).

LC-MS (Method 5): Rt=1.40 min; MS (ESIpos): m/z=211 [M+H]+.

 …………

RM 2

Example 16A 4-(6-Hydrazinopyrimidin-4-yl)morpholine

Figure US20100305085A1-20101202-C00043

Stage a):

4-(6-Chloropyrimidin-4-yl)morpholine

Figure US20100305085A1-20101202-C00044

45.0 g (302.1 mmol) 4,6-dichloropyrimidine are initially introduced into 450 ml water. 26.3 g (302.1 mmol) morpholine are added and the mixture is stirred at 90° C. for 16 h. Thereafter, it is cooled to 0° C. and the precipitate formed is filtered off. The precipitate is washed once with 50 ml water and dried in air.

Yield: 51.0 g (85% of th.)

LC-MS (Method 4): Rt=1.09 min; MS (ESIpos): m/z=200 [M+H]+;

1H-NMR (400 MHz, DMSO-d6): δ=8.35 (s, 1H), 6.95 (s, 1H), 3.62 (s, 8H).

Stage b)

4-(6-Hydrazinopyrimidin-4-yl)morpholine

Figure US20100305085A1-20101202-C00045

53.0 g (2.7 mmol) 4-(6-chloropyrimidin-4-yl)morpholine are initially introduced into 260 ml ethanol. 132.9 g (2.7 mol) hydrazine hydrate are added and the mixture is stirred under reflux for 16 h. Thereafter, it is cooled to RT and approx. half of the solvent is removed by distillation. The mixture is cooled to 0° C. and the solid formed is filtered off. It is rinsed with cold ethanol and the solid is dried first in air and then in vacuo.

Yield: 35.0 g (68% of th.)

LC-MS (Method 1): Rt=0.17 min; MS (ESIpos): m/z=196 [M+H]+;

1H-NMR (400 MHz, DMSO-d6): δ=7.94 (s, 1H), 7.70 (s, 1H), 5.91 (s, 1H), 4.15 (s, 2H), 3.66-3.60 (m, 4H), 3.45-3.37 (m, 4H).

 ………..

Example 71

2-(6-Morpholin-4-ylpyrimidin-4-yl)-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

Figure US20100305085A1-20101202-C00156

1.9 g (8.8 mmol) of the compound from Example 3A and 1.9 g (9.7 mmol) of the compound from Example 16A are initially introduced into 25 ml ethyl acetate and 504 mg (4.4 mmol) TFA are added at RT. The mixture is stirred under reflux for 16 h, then cooled to 5° C. and subsequently stirred for a further 2 h. The solid formed is filtered off, washed with ethyl acetate and dried first in air and thereafter under a high vacuum. 1.7 g of product are obtained.

The mother liquor is combined with the wash solution and the solvent is removed. According to LC-MS, the residue (2.4 g) still contains the intermediate 3-[2-(6-morpholin-4-ylpyrimidin-4-yl)hydrazino]-2-(1H-1,2,3-triazol-1-yl)prop-2-enoic acid ethyl ester (intermediate stage of the cyclization), which is used directly for the preparation of Example 72 (see there).

Yield: 1.7 g (61% of th.)

LC-MS (Method 9): Rt=0.90 min; MS (ESIpos): m/z=315 [M+H]+;

1H-NMR (400 MHz, DMSO-d6): δ=8.42 (s, 1H), 8.38 (s, 1H), 8.01 (s, 1H), 7.73 (s, 1H), 7.70 (s, 1H), 3.71-3.65 (m, 4H), 3.57-3.51 (m, 4H).

………..

Hydrochloride

Example 72

2-(6-Morpholin-4-ylpyrimidin-4-yl)-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one hydrochloride

Figure US20100305085A1-20101202-C00157

Batch 1: 7.5 ml of a 4 N solution of hydrogen chloride in dioxane are added to 1.7 g (5.4 mmol) of the compound from Example 71. The mixture is stirred at RT, 5 ml dioxane are added and the mixture is stirred at RT for 16 h. The solid is filtered off and washed with 5 ml dioxane. The mixture is dried under a high vacuum for 16 h, 10 ml methanol are then added and the mixture is stirred at RT for 1 h. The solid is filtered off, washed with 4 ml methanol and dried under a high vacuum. 1.6 g of the title compound are obtained.

Batch 2: A further amount of the title compound is obtained as follows: The residue (2.4 g) obtained from the mother liquor during the synthesis of Example Compound 71, which contains the open-ring intermediate state of the cyclization, 3-[2-(6-morpholin-4-ylpyrimidin-4-yl)hydrazino]-2-(1H-1,2,3-triazol-1-yl)prop-2-enoic acid ethyl ester, is dissolved in 12 ml ethanol and 1.5 ml 30% strength sodium methylate solution in methanol are added at RT, while stirring. The mixture is subsequently stirred at RT for 45 min, then adjusted to pH 5 with 2 N hydrochloric acid and subsequently stirred at RT for a further 16 h. The mixture is cooled to 10° C. and the solid is filtered off and washed with 3.5 ml dioxane. The mixture is dried under a high vacuum for 16 h, 5 ml methanol are then added and the mixture is subsequently stirred at RT for 1 h. The solid is filtered off, washed with 2 ml methanol and dried under a high vacuum to give a further 997 mg of the title compound in this way.

Yield: together 2.6 g (83% of th.)

LC-MS (Method 6): Rt=0.89 min; MS (ESIpos): m/z=315 [M+H]+;

1H-NMR (400 MHz, DMSO-d6): δ=8.54 (s, 1H), 8.39 (s, 1H), 8.28 (s, 1H), 7.88 (s, 1H), 7.42 (s, 1H), 3.71 (s, 8H).

References

  1. Jump up^ Flamme, I; Oehme, F; Ellinghaus, P; Jeske, M; Keldenich, J; Thuss, U (2014). “Mimicking hypoxia to treat anemia: HIF-stabilizer BAY 85-3934 (Molidustat) stimulates erythropoietin production without hypertensive effects”PLoS ONE9 (11): e111838. Bibcode:2014PLoSO…9k1838Fdoi:10.1371/journal.pone.0111838PMC 4230943PMID 25392999.
  2. Jump up^ Gupta, Nupur; Wish, Jay B (2017). “Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitors: A Potential New Treatment for Anemia in Patients with CKD”. American Journal of Kidney Diseases69 (6): 815. doi:10.1053/j.ajkd.2016.12.011PMID 28242135.
  3. Jump up^ Dib, Josef; Mongongu, Cynthia; Buisson, Corinne; Molina, Adeline; Schänzer, Wilhelm; Thuss, Uwe; Thevis, Mario (2017). “Mass spectrometric characterization of the hypoxia-inducible factor (HIF) stabilizer drug candidate BAY 85-3934 (molidustat) and its glucuronidated metabolite BAY-348, and their implementation into routine doping controls”. Drug Testing and Analysis9 (1): 61–67. doi:10.1002/dta.2011PMID 27346747.
Patent ID

Title

Submitted Date

Granted Date

US8653111 Substituted dihydropyrazolones for treating cardiovascular and hematological diseases
2012-01-23
2014-02-18
US8653074 Substituted sodium 1H-pyrazol-5-olate
2011-11-08
2014-02-18
US8389520 SUBSTITUTED DIHYDROPYRAZOLONES FOR TREATING CARDIOVASCULAR AND HEMATOLOGICAL DISEASES
2010-12-02
US2016015786 MOBILIZING AGENTS AND USES THEREFOR
2013-11-04
2016-01-21
US2015087827 METHOD FOR THE PREPARATION OF TRIAZOLE COMPOUNDS
2013-05-06
2015-03-26
Molidustat
Molidustat structure.png
Clinical data
Synonyms Bay 85-3934
ATC code
  • None
Identifiers
CAS Number
PubChem CID
UNII
Chemical and physical data
Formula C13H14N8O2
Molar mass 314.31 g·mol−1
3D model (JSmol)

//////////MolidustatBay 85-3934

BMS 986142


Image result for BMS-986142

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BMS-986142

(2S,5R,3S)-6-fluoro-5-(3-(8-fluoro-1-methyl-2,4-dioxo-1,4-dihydroquinazolin-3(2H)-yl)-2-methylphenyl)-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide

6-Fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2- methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide

Molecular Formula, C32-H30-F2-N4-O4, Molecular Weight, 572.609, RN: 1643368-58-4
UNII: PJX9GH268R

  • Originator Bristol-Myers Squibb
  • Class Anti-inflammatories; Antirheumatics; Small molecules
  • Mechanism of Action Agammaglobulinaemia tyrosine kinase inhibitors
  • Phase II Rheumatoid arthritis; Sjogren’s syndrome
  • 24 Jun 2018 Biomarkers information updated
  • 07 Jun 2018 Bristol-Myers Squibb completes a phase II trial in Rheumatoid arthritis (Treatment-experienced) in Argentina, Austria, Belgium, Brazil, Canada, Chile, Colombia, Czech Republic, France, Germany, Israel, Italy, Japan, Mexico, Netherlands, Poland, Russia, South Africa, South Korea, Spain, Taiwan, USA (PO) (NCT02638948) (EudraCT2015-002887-17)
  • 01 Oct 2016 Phase-II clinical trials in Sjogren’s syndrome in Puerto Rico (PO) (NCT02843659) after October 2016
  •  phase II clinical development at Bristol-Myers Squibb for the treatment of patients with moderate to severe rheumatoid arthritis and for the treatment of moderate to severe primary Sjogren’s syndrome.

BMS-986142 is a potent, selective, reversible BTK inhibitor. BMS-986142 shows BTK IC50 = 0.5nM; human WB IC50 = 90 nM. In molecule of BMS-986142, two atropisomeric centers were rotationally locked to provide a single, stable atropisomer, resulting in enhanced potency and selectivity as well as a reduction in safety liabilities. With significantly enhanced potency and selectivity, excellent in vivo properties and efficacy, and a very desirable tolerability and safety profile, BMS-986142 was advanced into clinical studies substituted tetrahydrocarbazole and 10 carbazole carboxamide compounds useful as kinase inhibitors, including the modulation of Bruton’s tyrosine kinase (Btk) and other Tec family kinases such as Itk. Provided herein are substituted tetrahydrocarbazole and carbazole carboxamide compounds, compositions comprising such compounds, and methods of their use. The invention further pertains to pharmaceutical compositions containing at least one compound 15 according to the invention that are useful for the treatment of conditions related to kinase modulation and methods of inhibiting the activity of kinases, including Btk and other Tec family kinases such as Itk, in a mammal. Protein kinases, the largest family of human enzymes, encompass well over 500 proteins. Btk is a member of the Tec family of tyrosine kinases, and is a regulator of 20 early B-cell development, as well as mature B-cell activation, signaling, and survival. B-cell signaling through the B-cell receptor (BCR) leads to a wide range of biological outputs, which in turn depend on the developmental stage of the B-cell. The magnitude and duration of BCR signals must be precisely regulated. Aberrant BCR- mediated signaling can cause disregulated B-cell activation and/or the formation of 25 pathogenic auto-antibodies leading to multiple autoimmune and/or inflammatory diseases. Mutation of Btk in humans results in X-linked agammaglobulinaemia (XLA). This disease is associated with the impaired maturation of B-cells, diminished immunoglobulin production, compromised T-cell-independent immune responses and marked attenuation of the sustained calcium signal upon BCR stimulation. 30 Evidence for the role of Btk in allergic disorders and/or autoimmune disease and/or inflammatory disease has been established in Btk-deficient mouse models. For example, in standard murine preclinical models of systemic lupus erythematosus (SLE), Btk deficiency has been shown to result in a marked amelioration of disease progression. Moreover, Btk deficient mice are also resistant to developing collagen-induced arthritis and are less susceptible to Staphylococcus-induced arthritis.

A large body of evidence supports the role of B-cells and the humoral immune system in the pathogenesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics (such as RITUXAN®) developed to deplete B-cells, represent an important approach to the treatment of a number of autoimmune and/or inflammatory diseases. Because of Btk’s role in B-cell activation, inhibitors of Btk can be useful as inhibitors of B-cell mediated pathogenic activity (such as autoantibody production).

Btk is also expressed in mast cells and monocytes and has been shown to be important for the function of these cells. For example, Btk deficiency in mice is associated with impaired IgE-mediated mast cell activation (marked diminution of TNF-alpha and other inflammatory cytokine release), and Btk deficiency in humans is associated with greatly reduced TNF-alpha production by activated monocytes.

Thus, inhibition of Btk activity can be useful for the treatment of allergic disorders and/or autoimmune and/or inflammatory diseases including, but not limited to: SLE, rheumatoid arthritis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, multiple sclerosis (MS), transplant rejection, type I diabetes, membranous nephritis, inflammatory bowel disease, autoimmune hemolytic anemia, autoimmune thyroiditis, cold and warm agglutinin diseases, Evans syndrome, hemolytic uremic syndrome/thrombotic thrombocytopenic purpura (HUS/TTP), sarcoidosis, Sj5gren’s syndrome, peripheral neuropathies (e.g., Guillain-Barre syndrome), pemphigus vulgaris, and asthma. In addition, Btk has been reported to play a role in controlling B-cell survival in certain B-cell cancers. For example, Btk has been shown to be important for the survival of BCR-Abl-positive B-cell acute lymphoblastic leukemia cells. Thus inhibition of Btk activity can be useful for the treatment of B-cell lymphoma and leukemia. In view of the numerous conditions that are contemplated to benefit by treatment involving modulation of protein kinases, it is immediately apparent that new compounds capable of modulating protein kinases such as Btk and methods of using these compounds should provide substantial therapeutic benefits to a wide variety of patients.

U.S. Patent No. 8,084,620 and WO 2011/159857 disclose tricyclic carboxamide compounds useful as kinase inhibitors, including the modulation of Btk and other Tec family kinases. There still remains a need for compounds useful as Btk inhibitors and yet having selectivity over Jak2 tyrosine kinase. Further, there still remains a need for compounds useful as Btk inhibitors that have selectivity over Jak2 tyrosine kinase and also have improved potency in the whole blood BCR-stimulated CD69 expression assay. Applicants have found potent compounds that have activity as Btk inhibitors. Further, applicants have found compounds that have activity as Btk inhibitors and are selective over Jak2 tyrosine kinase. Further still, applicants have found compounds that have activity as Btk inhibitors, are selective over Jak2 tyrosine kinase, and have improved potency in the whole blood BCR-stimulated CD69 expression assay. These compounds are provided to be useful as pharmaceuticals with desirable stability, bioavailability, therapeutic index, and toxicity values that are important to their drugability.

SYN

CLIP

Adventures in Atropisomerism: A Case Study from BMS – Not a Real Doctor

Dennis Hu

Scheme 2. Highlights from optimization of the first intermediate with axial chirality.

Image result for BMS-986142

Image result for BMS-986142

CLIP

https://cen.acs.org/pharmaceuticals/drug-development/Giving-atropisomers-another-chance/96/i33

Image result for BMS-986142

Yet another atropisomeric kinase inhibitor, of Bruton’s tyrosine kinase (BTK), currently being evaluated in Phase II clinical trials for rheumatoid arthritis, comes from Bristol Myers-Squibb. BMS-986142 contains one point-chiral center and two atropisomeric chiral axes, making it a diastereomeric compound with eight possible isomers. The less stable atropisomeric axis has a half-life on the order of hours to days, which means it can’t be heated above about 45 °C without the compound morphing. To keep the molecule from racemizing, the team had to design its synthetic routes and analysis with a close eye on temperature.

During the discovery stage, BMS analytical chemist Jun Dai and the team developed methods to analyze the compounds’ isomers. She estimates that the researchers screened at least twice as many separation methods for atropisomers as they would have for normal chiral compounds because of the atropisomers’ potential for temperature-dependent conversion. “It was challenging but rewarding,” she says.

To determine the proportion of early atropisomers with half-lives of minutes to hours, the team ran high-performance liquid chromatography analysis at low temperature, chilling the column with ice or cooling equipment. Isolating some atropisomeric compounds required researchers to use ice-bath cooling during fraction collection and even solvent evaporation. The medicinal chemistry route to BMS-986142 required three chiral column purifications to obtain a single diastereomer with the best binding properties (J. Chromatogr. A 2017, DOI: 10.1016/j.chroma.2017.01.016).

Process synthesis, however, generally isn’t amenable to column chromatography steps, which can take weeks to months on a large scale. “To be honest, when I first saw it, I really wasn’t sure how we were going to make it,” says BMS chemist Thomas Razler, who led the process chemistry efforts to scale-up BMS-986142.

The researchers say extensive knowledge sharing between medicinal, analytical, and process teams about the atropisomeric compound was key to the program’s success. The process team took advantage of the fact that the diastereomeric forms of BMS-986142 had very different solubility profiles, enabling the chemists to replace all chiral chromatography with simpler crystallization steps and produce more than 200 kg of a single enantiomer and diastereomer (Org. Lett. 2018, DOI: 10.1021/acs.orglett.8b01218).

Although the final molecule is stable as a solid, the team says that in solution, the risk of racemization is higher. Citing ongoing work in that area of development, Razler declined to elaborate on how the molecule behaves in its formulation but notes the team hopes to publish that information next year. The atropisomerism is still an issue, he says, but a fascinating one.

Paper

Organic Letters, 20(13), 3736-3740; 2018

Adventures in Atropisomerism: Total Synthesis of a Complex Active Pharmaceutical Ingredient with Two Chirality Axes

Chemical & Synthetic DevelopmentBristol-Myers Squibb Company1 Squibb Drive, New Brunswick, New Jersey 08901, United States
Org. Lett.201820 (13), pp 3736–3740
DOI: 10.1021/acs.orglett.8b01218
Abstract Image

A strategy to prepare compounds with multiple chirality axes, which has led to a concise total synthesis of compound 1A with complete stereocontrol, is reported.

Figure

Figure

https://pubs.acs.org/doi/suppl/10.1021/acs.orglett.8b01218/suppl_file/ol8b01218_si_001.pdf

(2S,5R)-6-fluoro-5-(3-(8-fluoro-1-methyl-2,4-dioxo-1,4- dihydroquinazolin-3(2H)-yl)-2-methylphenyl)-2-(2-hydroxypropan-2-yl)-2,3,4,9- tetrahydro-1H-carbazole-8-carboxamide (1A).

1H NMR (500 MHz, DMSO-d6) 10.78 (s, 1H), 8.07 (br. s., 1H), 7.95 (d, J=7.8 Hz, 1H), 7.72 (dd, J=14.2, 8.0 Hz, 1H), 7.56 (d, J=10.8 Hz, 1H), 7.45 (br. s., 1H), 7.42 – 7.36 (m, 1H), 7.34 (d, J=6.9 Hz, 1H), 7.34 – 7.31 (m, 1H), 7.29 (dd, J=7.5, 1.3 Hz, 1H), 4.17 (s, 1H), 3.73 (d, J=8.0 Hz, 3H), 2.91 (dd, J=16.8, 4.4 Hz, 1H), 2.48 – 2.37 (m, 1H), 1.98 – 1.89 (m, 2H), 1.87 (d, J=11.0 Hz, 1H), 1.76 (s, 3H), 1.59 (td, J=11.5, 4.1 Hz, 1H), 1.20 – 1.12 (m, 1H), 1.11 (s, 6H). 13C NMR (125.8 MHz, DMSO-d6) 168.2 (d, J=1.8 Hz, 1C), 160.1 (d, J=3.6 Hz, 1C), 151.9 (d, J=228.9 Hz, 1C), 150.5 (d, J=41.8 Hz, 1C), 148.7 (d, J=205.3 Hz, 1C), 139.2, 135.1, 135.0, 134.8, 131.4, 130.6, 130.0 (d, J=7.3 Hz, 1C), 128.5, 127.1 (d, J=4.5 Hz, 1C), 125.7, 124.3 (d, J=2.7 Hz, 1C), 123.6 (d, J=8.2 Hz, 1C), 123.0 (d, J=23.6 Hz, 1C), 120.8 (d, J=20.0 Hz, 1C), 118.4, 115.3 (d, J=7.3 Hz, 1C), 108.8 (d, J=5.4 Hz, 1C), 106.7 (d, J=28.2 Hz, 1C), 70.4, 45.4, 34.3 (d, J=14.5 Hz, 1C), 27.1, 26.8, 24.8, 24.7, 22.1, 14.5. mp 222-225 °C. IR (neat) 3487, 3418, 3375, 2967, 1651, 1394, 756 cm-1; HRMS (ESI) m/z: calcd for C32H30F2N4O4 [M+H]+ 573.2308, found 573.2312.

Chiral HPLC Analysis: Gradient: Complex Start % B: 0 7 Min. 55% 11 Min. 55% 14 Min. 100% Stop Time: 17 min Flow Rate: 1.5 ml/min Wavelength1: 225 Wavelength2: 256 Solvent Pair: S194/S195 (TFA) Solvent A: A1=0.05%TFA Water:ACN (95:5) S194 Solvent B: B1=0.05%TFA Water:ACN (5:95) S195 Column 1 : 1: Chiralcel OX-3R 3um 4.6 x 150 mm SN = OX3RCD-TE001 Oven Temperature: 50

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Adventures in Atropisomerism: Development of a Robust, Diastereoselective, Lithium-Catalyzed Atropisomer-Forming Active Pharmaceutical Ingredient Step

Chemical and Synthetic DevelopmentBristol-Myers Squibb CompanyOne Squibb Drive, New Brunswick, New Jersey08903, United States
Org. Process Res. Dev., Article ASAP
DOI: 10.1021/acs.oprd.8b00246
Abstract Image

The final step in the route to BMS-986142, a reversible inhibitor of the BTK enzyme, involves the diastereoselective construction of a chiral axis during the base-mediated cyclization of the quinazolinedione fragment. Optimization of the reaction to minimize formation of the undesired atropisomer led to the discovery that the amount of base and nature of the counterion play a vital role in the diastereoselectivity of the reaction. The highest diastereoselectivities were observed with a catalytic amount of LiOt-Bu. Development of a crystallization to selectively purge the undesired atropisomer is reported. Interestingly, ripening of the crystalline API was observed and further investigated, leading to a significant increase in the purity of the active pharmaceutical ingredient.

(2S,5R)-6-fluoro-5-(3-(8-fluoro-1-methyl-2,4-dioxo-1,4- dihydroquinazolin-3(2H)-yl)-2-methylphenyl)-2-(2-hydroxypropan-2-yl)-2,3,4,9- tetrahydro-1H-carbazole-8-carboxamide 1A

white crystalline solid (80.52g, 6 wt % MeOH, 89.4% corrected yield).

1H NMR (500 MHz, DMSO-d6) 10.78 (s, 1H), 8.07 (br. s., 1H), 7.95 (d, J=7.8 Hz, 1H), 7.72 (dd, J=14.2, 8.0 Hz, 1H), 7.56 (d, J=10.8 Hz, 1H), 7.45 (br. s., 1H), 7.42 – 7.36 (m, 1H), 7.34 (d, J=6.9 Hz, 1H), 7.34 – 7.31 (m, 1H), 7.29 (dd, J=7.5, 1.3 Hz, 1H), 4.17 (s, 1H), 3.73 (d, J=8.0 Hz, 3H), 2.91 (dd, J=16.8, 4.4 Hz, 1H), 2.48 – 2.37 (m, 1H), 1.98 – 1.89 (m, 2H), 1.87 (d, J=11.0 Hz, 1H), 1.76 (s, 3H), 1.59 (td, J=11.5, 4.1 Hz, 1H), 1.20 – 1.12 (m, 1H), 1.11 (s, 6H).

13C NMR (125.8 MHz, DMSO-d6) 168.2 (d, J=1.8 Hz, 1C), 160.1 (d, J=3.6 Hz, 1C), 151.9 (d, J=228.9 Hz, 1C), 150.5 (d, J=41.8 Hz, 1C), 148.7 (d, J=205.3 Hz, 1C), 139.2, 135.1, 135.0, 134.8, 131.4, 130.6, 130.0 (d, J=7.3 Hz, 1C), 128.5, 127.1 (d, J=4.5 Hz, 1C), 125.7, 124.3 (d, J=2.7 Hz, 1C), 123.6 (d, J=8.2 Hz, 1C), 123.0 (d, J=23.6 Hz, 1C), 120.8 (d, J=20.0 Hz, 1C), 118.4, 115.3 (d, J=7.3 Hz, 1C), 108.8 (d, J=5.4 Hz, 1C), 106.7 (d, J=28.2 Hz, 1C), 70.4, 45.4, 34.3 (d, J=14.5 Hz, 1C), 27.1, 26.8, 24.8, 24.7, 22.1, 14.5.

mp 222-225 °C.

IR (neat) 3487, 3418, 3375, 2967, 1651, 1394, 756 cm-1;

HRMS (ESI) m/z: calcd for C32H30F2N4O4 [M+H]+ 573.2308, found 573.2312.

Chiral HPLC Analysis: Gradient: Complex Start % B: 0 7 Min. 55% 11 Min. 55% 14 Min. 100% Stop Time: 17 min Flow Rate: 1.5 ml/min Wavelength1: 225 Wavelength2: 256 Solvent Pair: S194/S195 (TFA) Solvent A: A1=0.05%TFA Water:ACN (95:5) S194 Solvent B: B1=0.05%TFA Water:ACN (5:95) S195 Column 1 : 1: Chiralcel OX-3R 3um 4.6 x 150 mm SN = OX3RCD-TE001 Oven Temperature: 50…..https://pubs.acs.org/doi/suppl/10.1021/acs.oprd.8b00246/suppl_file/op8b00246_si_001.pdf

PAPER

Discovery of 6-Fluoro-5-(R)-(3-(S)-(8-fluoro-1-methyl-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (BMS-986142): A Reversible Inhibitor of Bruton’s Tyrosine Kinase (BTK) Conformationally Constrained by Two Locked Atropisomers

Bristol-Myers Squibb Research and Development, P.O. Box 4000, Princeton, New Jersey 08543, United States
J. Med. Chem.201659 (19), pp 9173–9200
DOI: 10.1021/acs.jmedchem.6b01088
Publication Date (Web): September 1, 2016
Copyright © 2016 American Chemical Society
*Phone: 609-252-6778. E-mail: scott.watterson@bms.com.
Abstract Image

Bruton’s tyrosine kinase (BTK), a nonreceptor tyrosine kinase, is a member of the Tec family of kinases. BTK plays an essential role in B cell receptor (BCR)-mediated signaling as well as Fcγ receptor signaling in monocytes and Fcε receptor signaling in mast cells and basophils, all of which have been implicated in the pathophysiology of autoimmune disease. As a result, inhibition of BTK is anticipated to provide an effective strategy for the clinical treatment of autoimmune diseases such as lupus and rheumatoid arthritis. This article details the structure–activity relationships (SAR) leading to a novel series of highly potent and selective carbazole and tetrahydrocarbazole based, reversible inhibitors of BTK. Of particular interest is that two atropisomeric centers were rotationally locked to provide a single, stable atropisomer, resulting in enhanced potency and selectivity as well as a reduction in safety liabilities. With significantly enhanced potency and selectivity, excellent in vivo properties and efficacy, and a very desirable tolerability and safety profile, 14f (BMS-986142) was advanced into clinical studies.

HPLC purity: 99.9%; tr = 11.05 min (Method A); 99.9%; tr = 10.72 min (Method B). Chiral purity: 99.8% ie;

Optical rotation: [α]D20 (c = 2.10, CHCl3) = +63.8°;

LCMS (ESI) m/z calcd for C32H30F2N4O4 [M + H]+ 573.2. Found: 573.5. Anal. calcd for C32H30F2N4O4, 0.72% H2O: C 65.56, H 5.42, N 9.55. Found: C 65.69, H 5.40, N 9.52.

 1H NMR (500 MHz, DMSO-d6) δ 10.78 (s, 1H), 8.07 (br. s., 1H), 7.95 (d, J = 7.8 Hz, 1H), 7.72 (dd, J = 14.2, 8.0 Hz, 1H), 7.56 (d, J = 10.8 Hz, 1H), 7.45 (br. s., 1H), 7.42–7.36 (m, 1H), 7.34 (d, J = 6.9 Hz, 1H), 7.34–7.31 (m, 1H), 7.29 (dd, J = 7.5, 1.3 Hz, 1H), 4.17 (s, 1H), 3.73 (d, J = 8.0 Hz, 3H), 2.91 (dd, J = 16.8, 4.4 Hz, 1H), 2.48–2.37 (m, 1H), 1.98–1.89 (m, 2H), 1.87 (d, J = 11.0 Hz, 1H), 1.76 (s, 3H), 1.59 (td, J = 11.5, 4.1 Hz, 1H), 1.20–1.12 (m, 1H), and 1.11 (s, 6H). 1

3C NMR (126 MHz, DMSO-d6) δ 168.2 (d, J = 1.8 Hz, 1C), 160.1 (d, J = 3.6 Hz, 1C), 151.9 (d, J = 228.9 Hz, 1C), 150.5 (d, J = 41.8 Hz, 1C), 148.7 (d, J= 205.3 Hz, 1C), 139.2, 135.1, 135.0, 134.8, 131.4, 130.6, 130.0 (d, J = 7.3 Hz, 1C), 128.5, 127.1 (d, J = 4.5 Hz, 1C), 125.7, 124.3 (d, J = 2.7 Hz, 1C), 123.6 (d, J = 8.2 Hz, 1C), 123.0 (d, J = 23.6 Hz, 1C), 120.8 (d, J = 20.0 Hz, 1C), 118.4, 115.3 (d, J = 7.3 Hz, 1C), 108.8 (d, J = 5.4 Hz, 1C), 106.7 (d, J = 28.2 Hz, 1C), 70.4, 45.4, 34.3 (d, J = 14.5 Hz, 1C), 27.1, 26.8, 24.8, 24.7, 22.1, and 14.5. 

19F-NMR (470 MHz, DMSO-d6) δ −121.49 (dt, J = 22.9, 11.4 Hz, 1F), and −129.56 (d, J = 11.4 Hz, 1F).

PATENT

WO 2014210085

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=850E1F706BE58D54C2B9AEE37AE6831C.wapp2nC?docId=WO2014210085&tab=PCTDESCRIPTION&queryString=EN_ALL%3Anmr+AND+PA%3A%28Bristol-Myers+Squibb%29+&recNum=19&maxRec=4726

Atropisomers are stereoisomers resulting from hindered rotation about a single bond axis where the rotational barrier is high enough to allow for the isolation of the individual rotational isomers. (LaPlante et al., J. Med. Chem., 54:7005-7022 (2011).)

Th compounds of Formula (A):

have two stereogenic axes: bond (a) between the tricyclic tetrahydrocarbazole/carbazole group and the phenyl group; and bond (b) between the asymmetric heterocyclic dione group Q and the phenyl group. Due to the non-symmetric nature of the substitutions on the rings connected by the single bonds labeled a and b, and due to limited rotation about these bonds caused by steric hindrance, the compounds of Formula (A) can form rotational isomers. If the rotational energy barriers are sufficiently high, hindered rotations about bond (a) and/or bond (b) occur at rates that are slow enough to allow isolation of the separated atropisomers as different compounds. Thus, the compounds of Formula (A) can form four rotational isomers, which under certain conditions, such as chromatography on a chiral stationary phase, can be separated into individual atropisomers. In solution, the compounds of Formula (A) can be provided as a mixture of four diastereomers, or mixtures of two pairs of diastereomers, or single atropisomers.

For the compounds of Formula (A), the pair of rotational isomers formed by hindered rotation about stereogenic axis (a) can be represented by the compounds of Formula (I) and Formula (B) having the structures:

The compounds of Formula (I) and the compounds of Formula (B) were found to be separable and stable in solution at ambient and physiological temperatures. Additionally, rotational isomers are formed by hindered rotation about stereogenic axis (b). These two atropisomers of the compounds of Formula (I) were also found to be separable and stable in solution at ambient and physiological temperatures.

Chiral compounds, such as the compounds of Formula (A), can be separated by various techniques including Supercritical Fluid Chromatography (SFC). SFC, which is form of normal phase HPLC, is a separation technique that uses super/subcritical fluid CO2 and polar organic modifiers such as alcohols as mobile phases. (White et al, J. Chromatography A, 1074: 175-185 (2005).

Example 28

6-Fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2- methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide (single atropisomer)


(28)

Following the procedure used to prepare Example 27, (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro- lH-carbazole-8-carboxamide (single enantiomer) [Intermediate 26] (0.045 g, 0.122 mmol) and 8-fluoro-l-methyl-3-(S)-(2-methyl-3-(4,4,5, 5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)quinazoline-2,4(lH,3H)-dione

[Intermediate 10] (0.065 g, 0.158 mmol) were converted into 6-fluoro-5-(3-(S)-(8-fluoro-1 -methyl-2,4-dioxo- 1 ,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-

hydroxypropan-2-yl)-2,3,4,9-tetrahydro- lH-carbazole-8-carboxamide (mixture of two atropisomers) as a yellow solid (0.035 g, 49% yield). Separation of a sample of this material by chiral super-critical fluid chromatography, using the conditions used to separate Example 27, provided (as the first peak to elute from the column) 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide. The chiral purity was determined to be greater than 99.5%. The relative and absolute configurations were determined by x-ray crystallography. Mass spectrum m/z 573 (M+H)+XH NMR (500 MHz, DMSO-d6) δ 10.77 (s, 1H), 8.05 (br. s., 1H), 7.94 (dd, J=7.9, 1.2 Hz, 1H), 7.56-7.52 (m, 1H), 7.43 (br. s., 1H), 7.40-7.36 (m, 1H), 7.35-7.30 (m, 2H), 7.28 (dd, J=7.5, 1.4 Hz, 1H), 4.15 (s, 1H), 3.75-3.70 (m, 3H), 2.90 (dd, J=16.8, 4.6 Hz, 1H), 2.47-2.39 (m, 1H), 1.93-1.82 (m, 3H), 1.74 (s, 3H), 1.57 (td, J=1 1.7, 4.2 Hz, 1H), 1.16-1.11 (m, 1H), and 1.10 (d, J=1.9 Hz, 6H). [a]D: +63.8° (c 2.1, CHC13). DSC melting point onset temperature = 202.9 °C (heating rate = 10 °C/min.).

The absolute configuration of Example 28 was confirmed by single crystal x-ray analysis of crystals prepared by dissolving the compound in excess methanol and slowly evaporating the solvent at room temperature to provide a di-methanol solvate (crystalline form M2-1). Unit cell dimensions: a = 9.24 A, b = 7.97 A, c = 22.12 A, a = 90.0°, β = 94.1°, γ = 90.0°; Space group: P2i; Molecules of Example 28/asymmetric unit: 1 ;

Volume/Number of molecules in the unit cell = 813 A3; Density (calculated) = 1.301 g/cm3. Fractional atomic coordinates at 173 K are given in Table 6, and a depiction of the structure is given in Figure 5.

Alternative Synthesis of Example 28:

A mixture of (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide [Intermediate 1 1] (5.00 g, 13.54 mmol), 8-fluoro-l-methyl-3-(S)-(2-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)quinazoline-2,4(lH,3H)-dione [Intermediate 10] (6.67 g, 16.25 mmol), tripotassium phosphate (2 M in water) (20.31 mL, 40.6 mmol), and tetrahydrofuran (25 mL) was subjected to 3 evacuate-fill cycles with nitrogen. The mixture was treated with l, l’-bis(di-/er/-butylphosphino)ferrocene palladium dichloride (0.441 g, 0.677 mmol) and the mixture was subjected to 2 more evacuate- fill cycles with nitrogen. The mixture was stirred at room temperature overnight, then was diluted with EtOAc, washed sequentially with water and brine, and dried and concentrated. The residue was purified by column chromatography on silica gel, eluting with EtOAc-hexanes (sequentially 50%, 62%, 75% and 85%), to provide 6-fluoro-5-(3-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3-(S)-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide as a white solid (6.58 g, 85% yield).

Material prepared by this method (40.03 g, 69.9 mmol) was separated by chiral super-critical fluid chromatography to give (2S, 5R)-6-fluoro-5-(3-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide. Further purification was achieved by suspending this material in methanol, sonicating for 5 min, collection of the solid by filtration, rinsing the collected solid with methanol and drying at room temperature under reduced pressure to give a white solid (22.0 g, 90% yield).

2R ANALOGUE

Example 27

6-Fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2- methylphenyl)-2-(R)-(2-hydroxypropan-2-yl)-2,3 ,4,9-tetrahydro- 1 H-carbazole-8- carboxamide (single atropisomer)

Preparation 27A: 6-Fluoro-5-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(R)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (mixture of 2 atropisomers)

A mixture of (R)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (single enantiomer) [Intermediate 25] (5.00 g, 13.5 mmol), 8-fluoro-l-methyl-3-(S)-(2-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl) quinazoline-2,4(lH,3H)-dione [Intermediate 10] (6.94 g, 16.9 mmol), 2 M aqueous K3PO4 (20.3 mL, 40.6 mmol) and THF (60 mL) was subjected to three evacuate-fill cycles with nitrogen. The mixture was treated with 1 , l’-bis(di-tert-butylphosphino) ferrocene palladium(II) chloride (441 mg, 677 μιηοΐ) and subjected to two more evacuate-fill cycles with nitrogen. The mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc, washed sequentially with water and brine, and dried and concentrated. The residue was purified by column chromatography on silica gel, eluting with EtOAc-hexanes (sequentially 50%, 62%, 75% and 85%), to give 6-fluoro-5-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(R)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (mixture of two atropisomers) as an off-white solid (6.77 g, 87% yield). Mass spectrum m/z 573 (M+H)+. ¾ NMR (500 MHz, DMSO-d6) δ 10.79-10.74 (m, 1H), 8.05 (br. s., 1H), 7.98-7.93 (m, 1H), 7.76-7.69 (m, 1H), 7.57-7.51 (m, 1H), 7.43 (br. s., 1H), 7.40-7.26 (m, 4H), 4.19-4.13 (m, 1H), 3.74-3.68 (m, 3H), 2.94-2.84 (m, 1H), 2.49-2.35 (m, 2H), 1.92-1.80 (m, 3H), 1.76-1.68 (m, 3H), 1.62-1.52 (m, 1H), and 1.12-1.06 (m, 6H).

Example 27:

A sample of 6-fluoro-5-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(R)-(2-hydroxypropan-2-yl)-2, 3,4,9-tetrahydro-lH-carbazole-8-carboxamide (mixture of two atropisomers) was separated by chiral super-critical fluid chromatography as follows: column: CHIRALPAK® AS-H (3 x 25 cm, 5 μιη); Mobile Phase: C02-MeOH (70:30) at 120 mL/min, 35 °C, 100 bar; sample preparation: 9 mg/mL in MeOH; injection: 1.7 mL. The first peak eluting from the column provided 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(R)-(2 -hydroxypropan-2-yl)-2, 3,4,9-tetrahydro-lH-carbazole-8-carboxamide. The chiral purity was determined to be greater than 99.5%. Mass spectrum m/z 573 (M+H)+XH NMR (500 MHz, DMSO-d6) δ 10.76 (s, 1H), 8.05 (br. s., 1H), 7.96 (d, J=7.8 Hz, 1H), 7.72 (ddd, J=14.3, 8.0, 1.2 Hz, 1H), 7.55 (d, J=10.8 Hz, 1H), 7.44 (br. s., 1H), 7.40-7.36 (m, 1H), 7.35-7.28 (m, 3H), 4.18 (s, 1H), 3.72

PATENT

WO 2018118830

https://patentscope.wipo.int/search/de/detail.jsf?docId=WO2018118830&tab=PCTDESCRIPTION&office=&prevFilter=%26fq%3DICF_M%3A%22C07D%22%26fq%3DPAF_M%3A%22BRISTOL-MYERS+SQUIBB+COMPANY%22&sortOption=Ver%C3%B6ffentlichungsdatum+ab&queryString=&recNum=1&maxRec=1018

The present invention generally relates to processes for preparing a

tetrahydrocarbazole carboxamide compound.

Protein kinases, the largest family of human enzymes, encompass well over 500 proteins. Btk is a member of the Tec family of tyrosine kinases, and is a regulator of early B-cell development, as well as mature B-cell activation, signaling, and survival.

B-cell signaling through the B-cell receptor (BCR) leads to a wide range of biological outputs, which in turn depend on the developmental stage of the B-cell. The magnitude and duration of BCR signals must be precisely regulated. Aberrant BCR-mediated signaling can cause disregulated B-cell activation and/or the formation of pathogenic auto-antibodies leading to multiple autoimmune and/or inflammatory diseases. Mutation of Btk in humans results in X-linked agammaglobulinaemia (XLA). This disease is associated with the impaired maturation of B-cells, diminished immunoglobulin production, compromised T-cell-independent immune responses and marked attenuation of the sustained calcium signal upon BCR stimulation.

Evidence for the role of Btk in allergic disorders and/or autoimmune disease and/or inflammatory disease has been established in Btk-deficient mouse models. For example, in standard murine preclinical models of systemic lupus erythematosus (SLE), Btk deficiency has been shown to result in a marked amelioration of disease progression. Moreover, Btk deficient mice are also resistant to developing collagen-induced arthritis and are less susceptible to Staphylococcus-induced arthritis.

A large body of evidence supports the role of B-cells and the humoral immune system in the pathogenesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics (such as Rituxan) developed to deplete B-cells, represent an important approach to the treatment of a number of autoimmune and/or inflammatory diseases. Because of Btk’s role in B-cell activation, inhibitors of Btk can be useful as inhibitors of B-cell mediated pathogenic activity (such as autoantibody production).

Btk is also expressed in mast cells and monocytes and has been shown to be important for the function of these cells. For example, Btk deficiency in mice is

associated with impaired IgE-mediated mast cell activation (marked diminution of TNF-alpha and other inflammatory cytokine release), and Btk deficiency in humans is associated with greatly reduced TNF-alpha production by activated monocytes.

Thus, inhibition of Btk activity can be useful for the treatment of allergic disorders and/or autoimmune and/or inflammatory diseases including, but not limited to: SLE, rheumatoid arthritis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, multiple sclerosis (MS), transplant rejection, type I diabetes, membranous nephritis, inflammatory bowel disease, autoimmune hemolytic anemia, autoimmune thyroiditis, cold and warm agglutinin diseases, Evan’s syndrome, hemolytic uremic syndrome/thrombotic thrombocytopenic purpura (HUS/TTP), sarcoidosis, Sjogren’s syndrome, peripheral neuropathies (e.g., Guillain-Barre syndrome), pemphigus vulgaris, and asthma.

In addition, Btk has been reported to play a role in controlling B-cell survival in certain B-cell cancers. For example, Btk has been shown to be important for the survival of BCR-Abl-positive B-cell acute lymphoblastic leukemia cells. Thus inhibition of Btk activity can be useful for the treatment of B-cell lymphoma and leukemia.

Atropisomers are stereoisomers resulting from hindered rotation about a single bond axis where the rotational barrier is high enough to allow for the isolation of the individual rotational isomers. (LaPlante et al., J. Med. Chem. 2011, 54, 7005-7022).

US Patent 9,334,290 discloses substituted tetrahydrocarbazole and carbazole compounds useful as Btk inhibitors, including 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide as Example 28. 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide, referred to herein as Compound 8, has two stereogenic axes:

(i) bond “a” between the tricyclic tetrahydrocarbazole/carbazole group and the phenyl group; and (ii) bond “b” between the substituted tetrahydroquinazolinedione group and the phenyl group. Compound 8 has non-symmetric substitutions on the rings connected by the single bonds labeled “a” and “b”, and limited rotation about these bonds caused by steric hindrance. As the rotational energy barriers are sufficiently high, hindered rotations about bond (a) and bond (b) occur at rates that are slow enough to allow isolation of Compound 8 and the other atropisomers of Compound 8 as four individual diastereomeric atropisomer compounds. These four rotational isomers can be separated by

chromatography on a stationary phase to provide chiral mixtures of two atropisomers or individual atropisomers.

US Patent 9,334,290 discloses a multistep synthesis process for preparing the Compound 8. This process is shown schematically in Figures 2-4. The disclosed process includes three chiral separations from racemic mixtures including (i) a chiral separation of a racemic mixture of chiral enantiomers (FIG.2); (ii) chiral separation of a mixture of atropisomers along bond “b” between the substituted tetrahydroquinazolinedione group and the phenyl group (FIG.3); and chiral separation of a mixture of atropisomers along bond “a” between the tricyclic tetrahydrocarbazole/carbazole group and the phenyl group (FIG.4). In each one of these chiral separations, the maximum yield of the desired enantiomer or atropisomer from the racemic mixture is 50%.

There are difficulties associated with the adaptation of this multistep synthesis disclosed in US Patent 9,334,290 to a larger scale synthesis, such as production in a pilot plant or a manufacturing plant for commercial production. Additionally, it is desired to have a process that provides higher yields and/or reduces waste.

Applicants have discovered a synthesis process for the preparation of Compound 8 that provides higher yields, reduces waste, and/or is adaptable to large scale manufacturing.

he invention is illustrated by reference to the accompanying drawing described below.

FIG.1 shows the stereoselective synthesis scheme for the preparation of 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide, Compound 8, according to the processes of second aspect, the third aspect, and the first aspect of the invention.

FIG.2 shows the synthesis scheme disclosed in US 9,334,290 for the preparation of (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide, Compound 5 (Intermediate 26 in US 9,334,290).

FIG.3 shows the synthesis scheme disclosed in US 9,334,290 for the preparation of 8-fluoro-l-methyl-3-(S)-(2-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) phenyl)quinazoline-2,4(lH,3H)-dione, Intermediate 10 in US 9,334,290.

FIG.4 shows the synthesis scheme disclosed in US 9,334,290 for the preparation of Compound 8 from the coupling reaction of 8-fluoro-l -methyl-3-(S)-(2-methyl-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) phenyl)quinazoline-2,4(lH,3H)-dione, Intermediate 10, and (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro- lH-carbazole-8-carboxamide, Compound 5, to provide a racemic mixture of Example 27 in US 9,334,290; and the chiral separation of Example 27 to provide Compound 8.

wherein R is Ci-8 alkyl or benzyl;

in the presence of:

(i) one or more bases selected from lithium bases, sodium bases, potassium bases, cesium bases, l,8-diazabicycloundec-7-ene, and 1,1,3,3-tetramethylguanidine; and

(ii) a solvent selected from n-butyl acetate (nBuOAc), cyclopentyl methyl ether (CPME), dimethoxy ethane (DME), dimethylacetamide (DMAc), dimethylformamide (DMF), 1,4-dioxane, ethyl acetate (EtOAc), isobutyl acetate (iBuOAc), isopropyl acetate (IP Ac), isopropyl alcohol (IP A), methanol (MeOH), methyl acetate (MeOAc), methyl isobutyl ketone (MIBK), N-methyl-2-pyrrolidone (NMP), 2-methyltetrahydrofuran (MeTHF), tetrahydrofuran (THF), tetrahydropyran (THP), and mixtures thereof;

to provide said Compound 8.

Intermediate Al

2-amino-4 robenzoic acid


(Al)

5% Pt/C (50% water-wet) (60 g, 6 wt%) was charged to a nitrogen blanketed vessel containing isopropyl acetate (22 L) and 4-bromo-5-fluoro-2-nitrobenzoic acid (1.00 kg, 3.79 mol). The headspace was exchanged three times with nitrogen and followed three times with hydrogen. The reaction mixture was stirred at 25 °C under an atmosphere of hydrogen. After 40 hours, the reaction was complete and the headspace was exchanged three times with nitrogen. The reaction mixture was filtered. The reaction vessel and filter train were rinsed with isopropyl acetate (5 L). The combined organic layers were concentrated under reduced pressure to 5.0 L. The solvent was then exchanged to toluene under reduced pressure and the resulting solids were isolated by filtration, washed with toluene, and dried at 50 °C under reduced pressure to afford 0.59 kg (66% yield) of 2-amino-4-bromo-5-fluorobenzoic acid as a white to off-white crystalline solid.

Additional 2-amino-4-bromo-5-fluorobenzoic acid was obtained by washing the spent catalyst twelve times with 2.75: 1 w/w THF in water (9.0 L). Each portion of wash was allowed to soak the spent catalyst for 30 minutes. The filtrate was concentrated to 10 L. The resulting solids were isolated by filtration, washed with water (1.0 L), and dried at 40 °C under reduced pressure to afford 0.15 kg (17% yield) of 2-amino-4-bromo-5-fluorobenzoic acid as an off-white crystalline solid. ¾ NMR (400 MHz, DMSO-de) δ 8.74 (br s, 2H), 7.50 (d, J=9.6 Hz, 1H), 7.08 (d, J=6.1 Hz, 1H). 13C NMR (101 MHz, DMSO-de) 5 168.2, 149.5, 148.8, 147.2, 119.9, 117.0, 116.8, 114.8, 114.6, 109.1.

HPLC Conditions: Column: Waters X-bridge C-18 (150X4.6mm, 3.5μ); Column

Temeprature: 30 °C; Solvent A: 0.05% TFA in water: acetonitrile (95:05 v/v); Solvent B: 0.05%TFA in water: acetonitrile:methanol (05:75:20 v/v); Diluent: 0.25 mg/ml in acetonitrile; Gradient: %B: 0 min. 5%; 20 min. 95%; 25 min. 95%; 26 min. 5%; stop time 30 min; Flow Rate: 0.8 ml/min; Wavelength: 230 nm; The retention time of 2-amino-4-bromo-5-fiuorobenzoic acid was 13.2 min. The retention time of 4-bromo-5-fluoro-2-nitrobenzoic acid was 12.9 min.

Intermediate A2

4-bromo-5-fluoro- -hydrazinylbenzoic acid hydrochloride

A solution of sodium nitrite (100.0 g, 6.38 mol) and water (1.8 L) was slowly charged to a cold slurry (0 °C) of 2-amino-4-bromo-5-fluorobenzoic acid (1.00 kg, 4.27 mol) in water (2.2 L) containing 35% HCl (2.1 kg, 20.15 mol). The reaction mixture slurry was stirred at 0 °C for 5 hours. The resultant cold diazonium salt slurry was charged over 4 hours to a cold solution (0 °C) of sodium bisulfite (2.66 kg, 25.0 mol in water (7.5 L). The diazonium reaction vessel was rinsed with cold water (2.5 L). The rinse water was transferred slowly to the reaction mixture. After 40 minutes, the reaction mixture was warmed to 20 °C over one hour. The reaction mixture slurry was stirred at 20 °C for 3 hours. After 3 hours, the reaction mixture was slowly transferred to a 60 °C solution of 35% HCl (15.0 kg, 144.0 mol) and water (3.0 L). The vessel was rinsed with water (2.5 L); and transferred to 35% HCl and water reaction mixture. The reaction mixture was stirred at 60 °C for 2 hours. The product was isolated by filtration and washed with water (3.0 L). The wet cake was charged back to the reactor and was

slurried with isopropyl acetate (9.0 L) for 1 hour at 20 °C. The product was isolated by filtration, washed with isopropyl acetate (1.0 L), and dried at 45-50 °C under reduced pressure to afford 0.99 kg (81 % yield) of 4-bromo-5-fluoro-2-hydrazinylbenzoic acid hydrochloride as an off-white crystalline solid in 95% purity. ¾ NMR (400 MHz, DMSO-de) δ 10.04 (br s, 3H), 9.00 (br s, 1H), 7.74 (d, J=9.1 Hz, 1H), 7.61 (d, J=5.8 Hz, 1H). 13C NMR (101 MHz, DMSO-de) δ 167.3, 153.0, 150.6, 144.5, 119.2, 1 18.0, 114.6. HPLC analysis: Column: Zorbax Eclipse Plus C 18 3.5 um, 150 x 4.6 mm ID; Column Temeprature: 30 °C; Solvent A: 10 mM ammonium formate in water:MeOH (90: 10 v/v); Solvent B: MeOH : ACN (70:30 v/v); Diluent: 50% CH3CN(aq); Gradient: %B: 0 min. 0%; 15 min. 90%; 18 min. 100%; stop time 18 min; Flow Rate: 1.0 ml/min; Wavelength: 240 nm. The retention time of the diazonium salt intermediate was 3.7 min. The retention time of the mono-sulfamic acid intermediate was 5.2 min. The retention time of 4-bromo-5-fluoro-2-hydrazinylbenzoic acid hydrochloride was 8.0 min. The retention time of 2-amino-4-bromo-5-fluorobenzoic acid was 8.7 min.

INTERMEDIATE Bl

(3-amino-2-methylphenyl)boronic acid hydrochloride

A 500 mL ChemGlass reactor (Reactor A) was equipped with mechanical stirrer and a nitrogen inlet. To the reactor was added 150 ml of methyl tetrahydrofuran. Next, Pd(OAc)2 (241 mg, 0.02 eq) was added, followed by the addition of P(o-tolyl)3 ligand (654 mg, 0.04 eq). The containers holding the Pd(OAc)2 and P(o-tolyl)3 were rinsed with 15 ml of methyl tetrahydrofuran, and the rinse solvents were added to the reactor. The reactor was sealed, evacuated to less than 150 mbar, and filled with nitrogen gas. This was repeated an additional four times to reduce the oxygen level to below 400 ppm. The reaction mixture was stirred for 30 min. Next, 10 g (1.0 eq) of 3-bromo-2-methyl aniline was charged to the inerted reactor. The container that held the 3-bromo-2-methyl aniline was rinsed with 15 ml of Me-THF and added into the reactor. KOAc (15.6 g, 3 eq) was added to the reactor. A slurry formed. The reaction mixture was inerted by using three vacuum/nitrogen cycles to an oxygen endpoint of less than 400 ppm.

A second 500 ml ChemGlass reactor was charged with 150 mL of MeOH, followed by the addition of 7.2 g (1.5 eq) of B2(OH)4. The resultant slurry was agitated at 25 °C. After 30 min, the B2(OH)4 was fully dissolved. The homogeneous solution was inerted by using 5 vacuum/nitrogen purge cycles to reduce the oxygen level to less than 400 ppm. The B2(OH)4/MeOH solution was transferred to Reactor A under a nitrogen atmosphere.

The reactor was inerted using three vacuum/nitrogen cycles with agitation to reduce the oxygen level to less than 400 ppm. The batch was heated to 50 °C (internal batch temperature). A slurry was observed when the temperature reached 40 °C. After reacting for 3 hrs, HPLC analysis of the reaction mixture showed 0.2 AP starting material remained. N-acetyl cysteine (2.0 g, 0.2 g/g) was added to Reactor A. The reaction mixture was stirred at 50 °C (internal batch temperature) for 30 min. The reaction stream was concentrated through distillation to 5 ml/g (~ 50 ml). Methyl tetrahydrofuran (200 ml, 20 ml/g) was charged to the slurry. The slurry was then concentrated via distillation to 150 ml (15 ml/g). Methyl tetrahydrofuran (150 ml, 15 ml/g) was charged to the reaction mixture. The slurry was cooled to 20 °C (batch temperature). Brine (26 wt%, 25 ml, 2.5 ml/g) was charged followed by the addition of aqueous Na2C03 (20 wt%, 15 ml, 1.5 ml/g). The reaction mass was agitated at a moderate rate (50~75/min) for 30 min. Celite (1 g, 0.1 g/g) was charged to the bi-phasic solution. The resultant slurry was agitated for 30 min. The slurry was filtered and transferred to Reactor B. The Celite cake was washed with 10 ml of methyl tetrahydrofuran. The bottom, lean aqueous phase was split from the organic phase and discarded. Brine (26 wt%, 25 ml, 2.5 ml/g) was charged followed by the addition of aqueous Na2C03 (20 wt%, 15 ml, 1.5 ml/g) to the organic solution. The resultant bi-phasic solution was agitated at a moderate rate (75 rpm) for 30 min. The bottom, lean aqueous phase was split from the organic phase and discarded. B2(OH)4 analysis of the rich organic solution did not detect B2(OH)4.

In Reactor B, the rich organic phase was concentrated via distillation to 50 ml (5 ml/g). The concentrated solution was cooled to 0-5 °C (batch temp). Concentrated HC1 (1.06 kg, 2.0 eq) was charged to the solution over 30 min with the batch temperature maintained below 10 °C. Once the concentrated HC1 was added, a slurry formed. The

slurry was agitated for 2 h at 5 °C. The slurry was filtered. The wet cake was washed with methyl tetrahydrofuran (2 X 20 ml). The cake was collected and dried at 50 °C under 100 mbar vacuum for 6 h to afford 8.4 g of 3-amino-2-methylphenyl)boronic acid hydrochloride as a white solid (83.5 % yield). ¾ NMR (500 MHz, D20) δ 7.48-7.23 (m, 3H), 4.78 (br s, 5 H); 2.32 (s, 3H). 13C NMR (126 MHz, D2O) δ 135.2, 134.7, 130.1, 128.0, 124.3, 17.4.

HPLC analysis: Column: Zorbax Eclipse Plus CI 8 3.5 um, 150 x 4.6 mm ID; Solvent A: 10 mM ammonium formate in water: MeOH=90: 10); Solvent B: CH3CN: MeOH (30:70 v/v); Gradient: % B: 0 Min. 0%; 1 Min. 0%; 15 Min. 90%; 15.1 Min. 0%; Stop Time: 20 min; Flow Rate: 1 ml/min; wavelength: 240 nm. The retention time of (3-amino-2-methylphenyl)boronic acid hydrochloride was 4.4 min. The retention time of (3-amino-2-methylphenyl)boronic acid hydrochloride was 17.8 min.

Intermediate CI

7-fluoro-l-methylindoline-2,3-dione

N,N-dimethylformamide (540.0 mL, 6980 mmol, 100 mass%) was added to a 2-L ChemGlass reactor equipped with a mechanical agitator, a temperature probe, and a cooling/heating circulator. Next, 7-fluoroindoline-2,3-dione (135.0 g, 817.6 mmol, 100 mass%) was added at 25 °C and dissolved to form a dark red solution. The charging ports and the beaker that contained the 7-fluoroindoline-2,3-dione were washed with N,N-dimethylformamide (135.0 mL, 1750 mmol, 100 mass%) and the rinse solution was poured into the reactor. Next, cesium carbonate 60-80 mesh (203.66 g, 625.05 mmol, 100 mass%) was added portion-wise to the reaction mixture. The addition was exothermic and the temperature of the reaction mixture increased from 20 to 25.5 °C. The color of the reaction mixture changed from a dark red solution to a black solution. The reactor jacket temperature was set to 0 °C. Next, iodomethane (56.5 mL, 907 mmol, 100 mass%) was added slowly via an additional funnel at ambient temperature, (iodomethane

temperature) while maintaining the batch temperature at less than 30 °C. Upon stirring, the reaction was exothermic, reaching a temperature of 29.3 °C. The batch temperature decreased to 26.3 °C after 85% of iodomethane was added, and the reaction mixture turned from black to an orange. After the addition of the iodomethane was completed, the jacket temperature was raised to 25.5 °C. The reaction mixture was stirred at 25 °C for 2 hrs.

The reddish orange-colored reaction mixture was transferred to a 1 L Erlenmeyer flask. The reaction mixture was filtered through a ceramic Buchner funnel with a No.1 Whatman filter paper to remove solid CS2CO3 and other solid by-products. In addition to a light-colored powder, there were yellow to brown colored rod-shaped crystals on top of the cake, which were water soluble. The filtrate was collected in a 2-L Erlenmeyer flask. The solids cake was washed with N,N-dimethylformamide (100.0 mL, 1290 mmol, 100 mass%). The DMF filtrate was collected in a 2-L Erlenmeyer flask.

To a separate 5-L ChemGlass reactor was charged water (3000.0 mL, 166530 mmol, 100 mass%). Next, 1.66 g of 7-fluoro-l-methylindoline-2,3-dione was added as seed to the water to form an orange colored suspension. The DMF filtrate was charged to the 5-L reactor slowly while maintaining the batch temp, at less than 29 °C over a period of 60 min. Stirring was maintained at 290 rpm. The orange solids precipitated instantly. The 2-L Erlenmeyer flask was rinsed with N,N-dimethylformamide (55.0 mL, 711 mmol, 100 mass%) and charged to the 5-L reactor. The slurry was cooled to 25 °C and agitated at 200 rpm for 12 hrs. The mixture remained as a bright orange-colored suspension. The slurry was filtered over a No. l Whatman filter paper in a 9 cm diameter ceramic Buchner funnel to a 4L Erlenmeyer flask to provide a bright orange-colored cake. The cake was washed with 1200 mL of water via rinsing the 5000 mL reactor (400 mL x 2), followed by 300 mL of deionized water introduced directly on the orange cake. The wet cake was dried under suction for 40 min at ambient temperature until liquid was not observed to be dripping from the cake. The cake was introduced into a vacuum oven (800 mbar) with nitrogen sweeping at ambient temperature for 1 hr, at 40-45 °C for overnight, and at 25 °C for 1 day to provide 7-fluoro-l-methylindoline-2,3-dione (Q, 130.02 g, 725.76 mmol, 100 mass%, 88.77% yield) as a bright orange-colored solid. ¾ NMR (400 MHz, DMSO-de) δ 7.57 (ddd, J=12.0, 8.5, 1.0 Hz, 1H), 7.40 (dd, J=7.3, 1.0 Hz, 1H), 7.12 (ddd, J=8.5, 7.5, 4.0 Hz, 1H), 3.29 (d, J=3.0 Hz, 3H). 13C NMR (101 MHz, DMSO-de) δ 182.3, 158.2, 148.8, 146.4, 137.2, 125.9, 124.3, 120.6, 28.7.

Intermediate C2

3-fluoro-2-(methylamino)benzoic acid

To a 1-L three neck round bottom flask equipped with a mechanical overhead agitator, a thermocouple, and an ice-water bath was charged NaOH (5.0 N) in water (140.0 mL, 700 mmol, 5.0 mol/L) followed by deionized water (140.0 mL, 7771 mmol, 100 mass%) to form a colorless transparent solution (T = 20.2 °C). 7-fluoro-l-methylindoline-2,3-dione (R, 25 g, 139.55 mmol, 100 mass%) was charged portion-wise while controlling the batch temperature at less than 24 °C with an ice-water bath to provide cooling. 7-fluoro-l-methylindoline-2,3-dione was charged and 50 mL of water was used to rinse off the charging funnel, the spatula, and the charging port. The reaction mixture was a thick yellow-green hazy suspension. The yellow-greenish suspension was cooled to 5.0 °C with an ice-water bath. The mixture was stirred for 15 min. Next, hydrogen peroxide (50% wt.) in water (11.0 mL, 179 mmol, 50 mass%) was charged to a 60 mL additional funnel with deionized (4.0 mL, 220 mmol, 100 mass%). The concentration of H2O2 post dilution was ~ 36.7%. The dilute hydrogen peroxide solution was added over a period of 11 minutes to the 1 L round bottom flask cooled with an ice-water bath and stirred at 350 rpm. The reaction mixture color was observed to become lighter in color and less viscous after 5 mL of the peroxide solution was added. After adding 10 mL of peroxide solution, the reaction mixture became clear with visible solids. At the end of addition, the reaction mixture was a green-tea colored transparent solution. The ice-water bath was removed (batch temperature was 16.6 °C), and the transparent, greenish yellow reaction mixture was allowed to warm to ambient temperature (21.0 °C), stirred for 1 hr.

After the reaction was complete, (1.0 hr), the reaction mixture was cooled to 4.3 °C with an ice-water bath. The reaction mixture was neutralized by the addition 6.0 N HCl (aq.) over a period of 3 hours to minimize foaming and the exotherm, resulting in the formation of a yellow-green suspension. The ice-bath was removed and the quenched reaction mixture was stirred at ambient temperature for 20 min. The yellow-green colored reaction mixture was transferred to a 2 L separatory funnel. Dichloromethane (300.0 mL, 4680 mmol, 100 mass%) was charged to the separatory funnel via rinsing the 1 L 3-necked round bottom flask. The separatory funnel was shaken vigorously, then allowed to settle (phase split was fast). Gas evolution was minor. The top aqueous layer was dark amber in color. The bottom dichloromethane layer was tea-green in color. The bottom rich dichloromethane layer was transferred to a clean 1 L Erlenmeyer flask. Next, the 1 L three necked round bottom flask was rinsed again with dichloromethane (200.0 mL, 3120 mmol, 100 mass%). The dichloromethane rinse was added to the separatory funnel. The separatory funnel was shaken vigorously and allowed to settle (phase split was fast). The top aqueous layer was amber in color (lighter); the bottom

dichloromethane layer was lighter green. The bottom rich dichloromethane layer was transferred to the 1 L Erlenmeyer flask. Dichloromethane (200.0 mL, 3120 mmol, 100 mass%) was charged to the separatory funnel and the separatory funnel was shaken vigorously. The contents were allowed to settle (phase split was fast). The bottom rich dichloromethane layer was transferred to the same 1 L Erlenmeyer flask. Peroxide test strip showed > 10 mg/Liter peroxide concentration. The total volume of the aqueous layer was 540 mL.

In a separate 250-mL Erlenmeyer flask was added sodium thiosulfate

pentahydrate (20.0 g, 80.6 mmol, 100 mass%) followed by deionized water (180.0 mL, 9992 mmol, 100 mass%) to form a colorless solution (10% wt. solution). The sodium thiosulfate solution was added to the combined dichloromethane rich solution in the 1 L Erlenmeyer flask. The contents of the flask were stirred vigorously for 10 hrs at ambient temperature. Peroxide strip did not detect the presence of peroxides in the bottom DCM layer. The top Na2S203 layer was amber in color, the bottom dichloromethane layer was much lighter in color, but was still amber in color. After 10 hrs, the mixture was transferred to a 1 L separatory funnel. The top aqueous layer was discarded.

The dichloromethane solution was washed with 150.0 mL of saturated brine solution. After phase split, the bottom rich dichloromethane layer was transferred to a 1 L flask. The dichloromethane solution was distilled to approximately 150 mL to obtain an amber-colored solution. Next, dichloromethane (120 mL, 1872 mmol, 100 mass%) was added and the mixture was heated to 35-40 °C to fully dissolve the solids. The amber solution was filtered through a 0.45 micron PTFE membrane Zap Cap filtration unit into a 1 L flask. The filtrate was transferred into a 3-neck 1 L round bottom flask fitted with a thermocouple, a heating mantle, a mechanical agitator, and a condenser with a nitrogen inlet. To the flask was charged dichloromethane (120 mL, 1872 mmol, 100 mass%) via rinsing the 1 L flask. The contents of the flask were concentrated under reduced pressure to approximately 140 mL to afford a yellow-green-colored suspension. The mixture was heated to 40.5 °C (refluxing) with stirring at 155 rpm to form a green-colored suspension with white solid pieces. After refluxing for 5 min, heptane (100.0 mL, 683 mmol, 100 mass%) was charged to the above mixture. The batch temperature dropped from 41.3 °C to 33.8 °C and the reaction mixture was a suspension. The mixture was heated to 45 °C. The mixture remained as a suspension with supernatant being amber with white solids. The refluxing was mild. After 36 minutes, (batch temp. = 43.8 °C), heptane (120.0 mL, 819 mmol, 100 mass%) was added to the mixture. The batch temperature dropped to 38.0 °C. The reaction mixture was a suspension. The mixture was heated to 40-45 °C and seeded with 0.3 g of 3-fluoro-2-(methylamino)benzoic acid. The reaction mixture remained as a suspension with supernatant being amber and solid pieces of white color. At t = 1 h 25 min (T = 45.4 °C) heptane (100.0 mL, 683 mmol, 100 mass%) was charged to the mixture causing the temperature to drop to 41.0 °C. At t = 2 h l3 min, (T = 45.6 °C) additional heptane (100.0 mL, 683 mmol, 100 mass%) was added to the mixture causing temperature to drop to 41.7 °C. At t = 3 h 07 min, (T = 45.5 °C), the heating was stopped. The mixture was allowed to cool to 20-25 °C under a nitrogen blanket. The suspension was agitated at ambient temperature for 12 hrs. The mixture was filtered using No.1 Whatman filter paper fitted in a ceramic Buchner funnel to a 1 L Erlenmeyer flask. The solids were observed to settle quickly. The mother liquor was green in color. The bottom half of the round bottom flask was coated with a thin dark amber or brown film, which was water soluble. The 1 L round bottom flask was washed with 150 mL of heptane, and then the heptane was used to wash the collected off-white-colored solid.

The filter cake was allowed to dry at ambient temperature with suction for 10 min., then dried in a vacuum oven with nitrogen sweeping at 45-50 °C for 4 hrs, followed by drying at ambient temperature for 10 hrs, with nitrogen sweeping. 3-fluoro-2-(methylamino)benzoic acid (16.1 g) was isolated in 68.1 % yield. ¾ NMR (400 MHz, DMSO-de) δ 7.61 (d, J=7.7 Hz, IH), 7.23 (dq, J=7.9, 1.6 Hz, IH), 6.57 (td, J=8.0, 4.4 Hz, IH), 3.02 (d, J=6.8 Hz, 4H). 13C NMR (101 MHz, DMSO-de) δ 169.5, 153.1, 150.7, 141.8, 141.7, 127.4, 127.4, 120.9, 120.7, 114.8, 114.7, 114.4, 114.3, 32.8.

Intermediate C3

3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic

A 20 L jacketed glass reactor with an overhead mechanical agitator, a

thermocouple, a nitrogen inlet, a glass baffle, and a condenser rinsed with 4 liters of dichloromethane followed by nitrogen sweeping through bottom valve overnight. To the reactor was charged 3-fluoro-2-(methylamino)benzoic acid (1004.7 g, 5939.7 mmol, 100 mass%) followed by dichloromethane (6000 mL, 93400 mmol, 99.8 mass%) to form an off-white-colored suspension. Next, cesium carbonate (1035.2 g, 3170 mmol, 99.9 mass%) was added followed the addition of water (6000 g, 333056 mmol, 99 mass%) at ambient temperature. The batch temperature rose from 17.0 °C to 29.6 °C prior to addition of the water. Gas evolution was observed during the water charging. The colorless biphasic mixture was stirred for 15 min. The batch temperature was approximately 18.8 °C. Next, n-propyl chloroformate (806.0 g, 6445.4 mmol, 98 mass%) was charged to an addition funnel. The reaction mixture was cooled to 15.0 °C with a glycol circulator. The n-propyl chloroformate was added from the addition funnel to the mixture while maintaining the batch temperature between 15.0 and 20.0 °C over 1 hr with stirring at 156 rpm. At the end of the addition, the batch temperature was 18.1 °C. The jacket temperature was increased to 20 °C. The white milky reaction mixture was agitated for 90 minutes.

The agitation was stopped and the reaction mixture was allowed to settle for phase split for 50 min. The hazy, bottom rich dichloromethane layer split from the aqueous layer and was transferred to a carboy. Next, 500 g of anhydrous Na2S04 (s) and 100 g of 60-200 mesh silica gel was added to the dichloromethane solution of 3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid in the carboy. The dichloromethane solution was allowed to dry overnight.

The dichloromethane solution containing the 3-fluoro-2-(methyl

(propoxycarbonyl)amino)benzoic acid was transferred from the carboy to a clean 20 L reactor via a 10 micron Cuno® in-line filter under vacuum to remove solid Na2S04 and silica gel. The carboy was rinsed with 1 liter x 2 of dichloromethane to remove residual solids. The dichloromethane was distilled off in the 20 L reactor with the jacket temperature set at 32 °C, the batch temperature at 15 °C, and vacuum set to 200-253 torr. At the end of distillation, the crude product was a thick light-amber-colored syrup. The solution was concentrated to 3 L of dichloromethane, and refilled with 3 L of dichloromethane each time to a final fill volume of 6 L. Next, 1 liter of dichloromethane was charged via vacuum to the residue in the 20-L reactor. The solution of 3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid became hazier. The solution was filtered using a Buchner funnel with a No.1 filter paper into a new carboy. The reactor was rinsed with 500 mL x 2 of dichloromethane and the rinse was filtered through the same Buchner funnel. All the filtrates were combined in a carboy and stored at the ambient temperature under nitrogen. Yellow-colored solids were observed to settle at the bottom of the carboy. The solution of 3-fluoro-2-(methyl (propoxycarbonyl)amino)benzoic acid in dichloromethane was transferred back to the clean 20-L reactor via vacuum and a 1 micron Cuno® in-line filter. The filtrate was still slightly hazy. The carboy was rinsed with 300 mL x 3 of dichloromethane and the rinses were transferred to the reactor via the 1 micron Cuno® filter. The reactor walls were rinsed with 500-mL of dichloromethane. The dichloromethane solution was concentrated by distillation under reduced pressure until the volume was less than 2.0 liters.

The temperature of the reactor jacket was lowered to 30 °C. The vacuum was broken and the reactor was filed with nitrogen. To the reactor was added 2 liters of cyclohexane followed by 5.0 g of 3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid crystalline seed. The seeds did not dissolve. The mixture was allowed to stir at 30 °C for 5-10 min to form a thick slurry. Additional cyclohexane (2.0 L) was added over 2 minutes. The jacket temperature was lowered to 25 °C. The mixture was allowed to stir for 40 min. Additional cyclohexane (2.0 L) was added over 2 minutes. The j acket temperature was lowered to 23 °C. The suspension was maintained at 23 °C for 60 min. Additional cyclohexane (2.0 L) was added over 2 minutes. The suspension was stirred for 20 min. The jacket temperature was lowered to 19.0 °C. The suspension was maintained at 19-21 °C for 10 hrs. The slurry settled well after overnight aging. A sample of the supernatant was obtained and assessed for the loss based on 9.5 L total volume. The slurry was filtered to collect solids via a ceramic Buchner funnel with a No. l Whatman filter paper. The solids were crystalline and white when dry. The wet cake was washed with cyclohexane (~ 2000 mL x 3) followed by drying for 10 min. The cake volume was 4933 cm3. The wet cake was transferred to four Pyrex glass trays for heated drying. The drying was continued in a vacuum oven at ~ 35-40 °C with nitrogen sweeping for 12 hrs to afford 1302.9 g of 3-fluoro-2-(methyl(propoxycarbonyl)amino) benzoic acid in 85.9 % yield. ¾ NMR (400 MHz, DMSO-de) (3: 1 mixture of rotamers) δ 13.2 (br s, 1H), 7.72-7.67 (m, 1H), 7.58-7.52 (m, 1H), 7.49-7.43 (m, 1H), 4.06-3.95 (m, 0.50H), 3.90 – 3.80 (m, 1.50H) 3.12 (s 0.75H), 3.12 (s 2.25H), 1.67 – 1.58 (m, 0.50H), 1.42 – 1.34 (m5 1.50H), 0.93 (t, J=7.5 Hz, 0.75H), 0.67 (t, J=7.5 Hz, 2.25H). 13C NMR (101 MHz, DMSO-de) (mixture of rotamers) δ 165.8, 159.0, 156.6, 154.3, 131.6, 131.0, 128.7, 128.6, 126.3, 1 19.9, 119.7, 66.6, 66.4, 36.9, 36.4, 36.4, 21.8, 21.5, 10.0, 9.8.

HPLC Analysis: Column: Agilent ZORBAX Eclipse Plus C18 3.5um 4.6X150 mm; Column Temeprature: 40 °C; Solvent A: 0.01M NH4OOCH in water:MeOH (90: 10 v/v); Solvent B: O.OIM NH4OOCH in MeOH:CH3CN (70:30 v/v); Diluent: 0.25 mg/ml in acetonitrile; Gradient: %B: 0 min. 10%; 10 min. 30%; 20 min. 90%; 20.1 min. 10%; stop time 25 min; Flow Rate: 1.0 ml/min; Wavelength: 220 nm;

The retention time of 7-fluoro-l-methylindoline-2,3-dione was 10.7 minutes.

The retention time of 7-fluoroindoline-2,3-dione was 6.8 minutes. The retention time of 3-fluoro-2-(methylamino)benzoic acid was 5.9 minutes. The retention time of 3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid was 12.0 minutes.

Compound 1

(S)-3-(prop-l -en-2-yl)cyclohexan-l-one

Catalyst Preparation: Rhodium (I) (S)-(+)-5,5′-bis[di(3,5-di-tert-butyl-4-methoxyphenyl) phosphino] -4,4′-bi- 1 ,3-benzodioxole

Methanol (320 mL) was charged into a 0.5 L inerted reactor equipped with an overhead agitator, nitrogen sparging tube and an outlet connected to an oxygen meter. The reactor was inerted by sparging nitrogen subsurface through methanol until <300 ppm 02 was detected in the headspace. S-(+) DTBM-SEGPHOS (77.3 g, 65.6 mmol) and [Rh(cod)Cl]2 (15.4 g, 31 mmol) were charged and the nitrogen sparging continued until <300 ppm C was detected in the headspace. The mixture was agitated at room temperature under constant positive nitrogen pressure for 30 min by sweeping a low flow of nitrogen through the headspace. The initial yellow slurry gradually transformed into a deep-red solution containing a small amount of solids (excess ligand). The ligation completion was confirmed by 1P NMR by disappearance of the ligand peak at 13.1 ppm (s) and the appearance of the new singlets at 26.10 ppm and 27.01 ppm for the ligated species.

Synthesis of the Compound I

A 20 L jacketed Chemglass reactor, equipped with an overhead agitator, a thermocouple, nitrogen sparging tube, a sampling port, a condenser connected to the glycol supply and a nitrogen outlet connected sequentially to a bubbler, flow meter and an oxygen meter, was inerted using a vigorous nitrogen sweep. A Teledyne 3110 oxygen meter was used to monitor the progress of inertion. A vigorous nitrogen sweep was implemented prior to reagent charges until the oxygen reading was <300 ppm.

Heptane (4.0 L), 2-cyclohexen-l-one (1 kg, 10.4 M) in heptane (1.0 L), isopropenyl pinacol boronate (1.92 kg, 11.4 M, 1.1 eq) in heptane (1.0 L), DIPEA (0.91 L, 0.67 kg, 0.50 eq), a solution of 2,2-dimethy 1-1, 3 -propanediol (1.19 kg, 1.1 eq) in methanol (0.12L) in water (3 L), and additional heptane (2.55L) were sequentially charged to the reactor via vacuum. Nitrogen sparging subsurface through the agitated bi phasic mixture continued after the charges until an oxygen level of <300 ppm was

reached in the headspace prior to the catalyst charge. Then the nitrogen flow was reduced to maintain a slight positive pressure in the reactor.

The catalyst light slurry was transferred from the bottom value of the 0.5 L reactor’s bottom into the 20 L reactor through an inerted Teflon tubing by applying slight positive pressure of nitrogen. The contents of the small reactor was transferred including the excess of the undissolved solid.

The jacket was set to 60 °C on the 20 L reactor and the biphasic mixture was vigorously heated and agitated under nitrogen at 55-58 °C. After the transfer, the nitrogen flow was reduced to maintain a slight positive pressure and to minimize solvent loss. After completion of the reaction, the reaction mixture was cooled to 20-25 °C. The phases were separated and the organic phase was washed with IN HC1 aq (v=5.7 L, 0.55 eq) to remove DIPEA, and with water (2.5 L). Two back-extractions with heptane (2 x 2L) from the original aqueous phase were performed to bring back an additional 8 mol% of the product. All organic phases were combined and polished filtered back to the cleaned reactor. Heptane was removed under reduced pressure (30-40 °C at 45-55 torr) to give the crude product, which was transferred to a 2 L 4-necked round bottom flask, equipped with a mechanical stirrer, a thermocouple, a 30 cm Vigreaux column, a distillation adapter containing a thermocouple to measure the vapor temperature, a condenser (glycol) and a Teflon tubing attached to a receiver flask. Distillation was performed at a pressure of 10 torr with the main fraction containing the product boiling at 85-92 °C to afford 1.18 kg (85 mol % as is, 82.1 % corrected) of (S)-3-(prop-l-en-2-yl)cyclohexan-l-one. Chiral GC: Supelco AlphaDex 120 30 x 0.25 mm x 0.25 μπι, inlet 200 °C, split ratio 30: 1, carrier gas: helium, constant flow 1.9 mL/min, oven program: 80 °C to 110 °C at 2 °C /min, then 20 °C /min to 220 °C, detector: FID 250 °C; RT for the desired product: 14.4 min. Chemical purity: 97.1 GCAP. Chiral purity: ee = 99.6 %. ¾ NMR (CDCh): 1.57-1.70 (m, 12H), 1.75 (s, 3H), 1.91-1.96 (m, 1H), 2.05-2.12 (m, 1H), 2.26-2.46 (m, 5H), 4.73 (s, 1H), 4.78 (s, 1H).

Compound 2

(S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l-en-2-yl)cyclohexylidene)hydrazinyl)benzoic acid 

(S)-3 -(prop- l -en-2-yl)cyclohexan-l -one (50.00 mL, 33.4 mmol, 0.667 mmol/mL) solution in heptane was added to a Chemglass reactor. Next, 75 mL of MeOH was added. The MeOH solution was distilled at 60 torr/50 °C jacket temperature and 75 mL of constant volume with the addition of 300 mL of MeOH. The contents of the reactor were cooled to 20 °C. 2-amino-4-bromo-5-fluorobenzoic acid (8.5415 g, 29.918 mmol) was added to the reactor. The reaction mixture was stirred at 20 °C. After, 30 minutes, the solid material was dissolved to form a clear brown solution. After 2.0 h, water (25.0 mL) was added over 25 min to the reaction mixture under slow agitation (RPM = 100). After an additional 1.0 h, the slurry was filtered (fast; < 3 seconds). The cake was washed with 2×25 mL of MeOH/H20 (3:2). The cake was dried at 55 °C under vacuum overnight to afford (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l -en-2-yl)cyclohexylidene)

hydrazinyl)benzoic acid (10.5701 g; 95.7% yield). HPLC method: Column: Zorbax Eclipse plus 1.8 um C8 (4.6 X 50 mm); inj ection volume: 10 μί; Mobile Phase A: 0.05% TFA in acetonitrile: water (5 :95, v/v); Mobile Phase B: 0.05% TFA in water: acetonitrile (5:95, v/v); Gradient (%B) 0 min (30%), 14 min (100%), 15 min (30%); Flow Rate: 1.0 mL/min; Wavelength: 240 nm for IPC; Column temp: 25 °C; IPC Sample Prep:

Dissolved 10 of the reaction mixture and dilute with MeOH to 1.5 mL; HPLC results: Intermediate A2, 0.87 min; Compound 2, 9.97 min. ¾ NMR (400 MHz, DMSO-de) δ 13.54 (s, 1H), 10.76 (d, J = 26.5 Hz, 1H), 7.73 (appt triplet, J = 6.32 Hz, 1H), 7.64 (dd, J = 9.35, 1.26 Hz, 1H), 4.77-4.75 (m, 2H), 2.68-2.61 (m, 1H), 2.46-2.44 (m, 1H), 2.27-2.12 (m, 2H), 2.06-1.97 (m, 1H), 1.96-1.86 (m, 1H), 1.82-1.80 (m, 1H), 1.75-1.74 (m, 3H), 1.50-1.41 (m, 2H). 13C NMR (100 MHz, DMSO-de) δ 168.67, 152.76, 152.73, 150.71 , 148.41 , 148.38, 148.20, 145.10, 117.45, 117.21 , 116.45, 1 16.40, 1 15.76, 1 15.74, 1 15.54, 1 15.52, 109.64, 109.39, 108.88, 108.85, 108.83, 108.80, 44.80, 43.72, 34.22, 30.89, 30.08, 30.05, 25.42, 25.39, 24.15, 20.60, 20.44.

Compound 3

(S)-5-bromo-6-fluoro-2-(prop-l-en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxylic acid

Zinc chloride (8.7858 g, 64.46 mmol) and (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop- 1- en-2-yl)cyclohexylidene)hydrazinyl)benzoic acid (17.0011 g, 46.05 mmol) were added to a Chemglass reactor. Next, isopropyl acetate (170 mL) was added. The contents of the reactor were heated at 69.5 °C for 71 h and then cooled to room temperature. 2-MeTHF (205 mL) and HC1 (1 mol/L) in water (85 mL) were added. The reaction mixture was stirred at room temperature for 0.5 h. The layers were allowed to separate. The organic layer was washed with water (85 mL). The layers were separated and the organic layer was polish-filtered. The rich organic layer was distilled at 220 torr and 70 °C jacket temperature to 85 mL (5.0 mL/g (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l-en-2-yl)cyclohexylidene)hydrazinyl) benzoic acid). Next, the solution was distilled at 120 mL (7.0 mL/g (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l-en-2-yl)cyclohexylidene)hydrazinyl) benzoic acid) constant volume under 220 torr and 70 °C jacket temperature with continuous addition of acetonitrile (350 mL, 20 mL/g). Additional CFbCN was added to make the slurry volume = 153 mL (9.0 mL/g (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l-en- 2- yl)cyclohexylidene) hydrazinyl)benzoic acid). The slurry was heated to 82 °C batch temperature. After 3.0 h, the slurry was cooled to 20 °C over 2.0 h. The slurry was stirred at 20 °C for an additional 14 h. The slurry was filtered and the cake was washed with acetonitrile (2 x 17 mL, 1.0 mL/g (S,E)-4-bromo-5-fluoro-2-(2-(3-(prop-l-en-2-yl)cyclohexylidene) hydrazinyl)benzoic acid). The wet cake was dried in a vacuum oven at a temperature range of 50-55 °C overnight to afford (S)-5-bromo-6-fluoro-2-(prop-l-en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxylic acid (7.8991 g; 48.7% yield). HPLC method: Column: Agilent Zorbax Eclipse plus 1.8 μπι C8 (4.6 X 50 mm);

Injection Volume: 10 μί; Mobile Phase A: 0.05% TFA in acetonitrile: water (5:95, v/v); Mobile Phase B: 0.05% TFA in water: acetonitrile (5:95, v/v); Gradient (%B) 0 min

(30%), 14 min (100%), 15 min (100%); Flow Rate: 1.0 mL/min; Wavelength: 240 nm for IPC and Isolated product; Column temp: 25 °C; IPC Sample Prep: 1 mL/100 mL in tetrahydrofuran; Isolated Sample Prep: 0.25 mg/mL in tetrahydrofuran; HPLC results: Compound 3, 8.86 min; Compound 2, 10.0 min. ¾ NMR (400 MHz, DMSO-de) δ 13.41 (s, 1H), 11.03 (s, 1H), 7.45 (d, J = 9.85 Hz, 1H), 4.79 (appt d, J = 4.55Hz, 2H), 3.21-3.17 (m, 1H), 2.95 (dd, J = 17.18, 4.80 Hz, 1H), 2.91-2.83 (m, 1H), 2.61 (dd, J = 16.93, 10.61 Hz, 1H), 2.41-2.35 (m, 1H), 2.01-1.95 (m, 1H), 1.79 (s, 3H), 1.67-1.57 (m, 1H). 13C NMR (100 MHz, DMSO-de) δ 166.64, 166.61, 152.72, 150.42, 148.44, 139.96, 131.90, 127.44, 127.43, 112.40, 112.33, 109.67, 109.54, 109.39, 109.19, 109.14, 28.28, 27.79, 22.20, 20.69.

Compound 4

(S)-5-bromo-6-fluoro-2-(prop- -en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide

Acetonitrile (70 mL) was added to a Chemglass reactor, followed by the addition of (S)-5-bromo-6-fluoro-2-(prop-l-en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxylic acid (7.0150 g). Next, Ι,Γ-carbonyldiimidazole (4.2165 g, 26.004 mmol) was added. The reaction mixture was stirred (RPM = 100) for 5.0 hr at 20 °C. The slurry was cooled to 3 °C. Ammonia (30 mL, 200 mmol, 30 mass%) was added in less than 2 min. The slurry was stirred at 3 °C for 17.5 h. Water (70 mL) was added over 5 min. The slurry was stirred at 3 °C for 3 h. The slurry was filtered and the wet cake was washed with 2×50 mL of CH3CN/H2O (1 : 1). The wet cake was dried at 55 °C under vacuum overnight to afford (S)-5-bromo-6-fluoro-2-(prop-l-en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (5.2941 g; 75.8% yield). HPLC Method; Column: Agilent Zorbax Eclipse plus 1.8 μιη C8 (4.6 X 50 mm); Injection Volume: 10 μί; Mobile Phase A: 0.05% TFA in acetonitrile: water (5:95, v/v); Mobile Phase B: 0.05% TFA in water: acetonitrile (5:95, v/v); Gradient (%B) 0 min (0%), 8 min (100%), 10 min (100%); Flow Rate: 1.0 mL/min; Wavelength: 240 nm for IPC and Isolated product; Column temp: 25 °C; IPC Sample

Prep: Dissolved 10 of the reaction mixture into 1.0 mL 0.05 v% DBU/MeOH;

Product sample preparation: Dissolved product in MeOH at 1 mg/mL; HPLC results: Compound 4, 6.39 min; Compound 3, 6.80 min. ¾ NMR (400 MHz, DMSO-de) δ 11.05 (s, 1H), 8.11 (s, 1H), 7.59 (d, J = 10.36 Hz, 1H), 7.55 (br s, 1H), 4.78 (br s, 2H), 3.18 (br d, J = 14.65 Hz, 1H), 2.94 (dd, J = 16.93, 4.80 Hz, 1H), 2.88-2.82 (m, 1H), 2.62 (dd, J = 16.93, 10.61 Hz, 1H), 2.40-2.34 (m, 1H), 1.98 (d, J = 11.87 Hz, 1H), 1.78 (s, 3H), 1.66-1.56 (m, 1H). 13C NMR (100 MHz, DMSO-de) δ 167.64, 152.68, 150.38, 148.47, 139.47, 131.71, 127.02, 127.01, 115.36, 115.28, 109.53, 108.66, 108.61, 107.47, 107.19, 28.24, 27.87, 22.21, 20.67.

Compound 5

(S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide

Dichloromethane (100 mL) and (S)-5-bromo-6-fluoro-2-(prop-l-en-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (PPP, 10.0016 g, 28.48 mmol) were added to a 250 mL Chemglass reactor. The slurry was cooled to 5 °C. Next, trifluoroacetic acid (14.68 g, 128.7 mmol) was added over 0.5 h with agitation (RPM = 250) while maintaining the internal temperature at less than 10 °C). The temperature was raised to 14 °C and the reaction mixture was stirred at 14 °C for 17.5 h. Next, 60 mL of MeOH was added to dissolve the thin slurry. The solution was cooled to -10 °C. The solution was distilled at 80 torr while the jacket temperature was gradually raised from -10 °C to 20 °C. The solution was distilled to about 60 mL volume. The internal temperature changed from -7 °C to -2 °C. The solution became a heavy slurry. The distillation was continued at 80 torr at 20 °C jacket temperature at 60 mL volume with the addition of 120 mL MeOH. The intemal temperature changed from -2 °C to 15 °C. The solution became a heavy slurry. The distillation became slow. The vacuum pressure was changed to 60 torr, and the distillation was continued with a 20 °C jacket temperature to 40 mL slurry volume. The batch temperature went from 12 °C to 13 °C.

MeOH (20 mL) was sprayed to wash solid crust off the reactor wall, but was not effective. Aqueous N¾ (30.0 mL, 400 mmol, 28 mass%) was sprayed to the slurry (pH = 10.59). Some solid crust on the upper reactor wall still remained. The slurry was stirred at 20 °C for 0.5 h (pH = 10.58), then heated to 70 °C in 15 min. All the solid crust on the upper reactor wall dissolved. Next, water (40 mL) was added over a period of 15 min. The solution remained as a clear solution at 70 °C.

The slurry was seeded with solid (S)-5-bromo-6-fluoro-2-(2 -hydroxy propan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (~ 5 mg). The seeds remained but there was little additional crystallization was observed at 70 °C. The slurry was heated at 70 °C (jacket temperature = 80 °C) for 0.5 h, and then cooled down to 20 °C in 0.5 h. At 65 °C the mixture became cloudy. The mixture was stirred at 20 °C for 65 h. The mixture was filtered. The cake was washed with 2×15 mL of MeOH/LhO (1 : 1). The wet cake was dried at 65 °C under vacuum for 24 h, giving (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (9.1741 g, 87.3% yield).

(S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide was recrystallization in MeOH/MTBE/n-Heptane (1 :4:8).

(S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (8.0123 g) was added to a reactor. Next, MeOH (8.0 mL) and MTBE (32.0 mL) were added. The mixture was heated to 45 °C to dissolve the slurry. Heptane (64 mL) was added over a period of 15 min at 45 °C. The slurry was stirred at 45 °C for an additional 0.5 h and then cooled to 5 °C in 1.0 h. Stirring was continued at 5 °C for an additional 1.0 h. The slurry was filtered and the wet cake was washed with 2×20 mL of n-heptane. The wet cake was dried at 65 °C under vacuum for 16 h to afford (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (6.9541 g; 86.8%).

(S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (8.0123 g) was added to a reactor. Next, MeOH (8.0 mL) and MTBE (32.0 mL) were added. The mixture was heated to 45 °C to dissolve the slurry. Heptane (64 mL) was added over a period of 15 min at 45 °C. The slurry was stirred at 45 °C for an additional 0.5 h and then cooled to 5 °C in 1.0 h. Stirring was continued at 5 °C for an additional 1.0 h. The slurry was filtered and the wet cake was washed with 2×20 mL of n-heptane. The wet cake was dried at 65 °C under vacuum for 16 h to afford (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (6.9541 g; 86.8%). HPLC method Column: Phenomenex Kinetex C18 2.6um 100A 4.6X150mm SN:538219-97; Injection Volume 5 μί; Mobile Phase A: 0.05% TFA in acetonitrile:water (5:95, v/v); Mobile Phase B: 0.05% TFA in

water: acetonitrile (5 :95, v/v); Gradient (%B) 0 min (32%), 5 min (38%), 1 1 min (38%), 18 min (68%), 22 min (68%), 30 min (90%), 31 min (100%); Flow Rate: 1.0 mL/min; Wavelength: 220 nm for IPC and Isolated product; Column temp: 25 °C; IPC Sample Prep: 1 μΙ71 mL in tetrahydrofuran; Isolated Sample Prep: 0.25 mg/mL in

tetrahydrofuran; HPLC results: Compound 5, 9.58 min; Compound 4, 19.98 min; ¾ NMR (400 MHz, DMSO-de) δ 10.99 (s, 1H), 8.10 (s, 1H), 7.57 (d, J = 10.36 Hz, 1H), 7.54 (br s, 1H), 4.27 (s, 1H), 3.26 (dd, J = 15.66, 4.29 Hz, 1H), 2.93 (dd, J = 17.18, 4.55 Hz, 1H), 2.76-2.68 (m, 1H), 2.44 (dd, J = 16.17, 1 1.87 Hz, 1H), 2.12 (br d, J = 1 1.12 Hz, 1H), 1.69-1.62 (m, 1H), 1.31 (ddd, J = 25.01, 12.38, 5.31 Hz, 1H), 1.14 (s, 6H). 13C

NMR (100 MHz, DMSO-de) δ 167.67, 152.64, 150.34, 140.46, 131.77, 127.03, 127.02, 1 15.28, 1 15.21, 109.09, 109.05, 107.30, 107.03, 101.43, 101.19, 70.37, 44.96, 27.17, 26.73, 24.88, 24.36, 22.85.

Compound 6

(2S)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro- lH-carbazole-8-carboxamide

Catalyst activation

Into a 1 Liter Chemglass reactor (Reactor A) were added Me-THF (4 L/kg) followed by (R)-BINAP (0.0550 mol/mol, 7.45 mmol) and Pd(OAc)2 (0.0500 mol/mol, 6.77 mmol). Additional Me-THF (1 L/kg) was added. The mixture was stirred at 25 °C

for 1 h. Next, 4-bromo-3-fluoro-7-(l-hydroxy-l-methyl-ethyl)-6,7,8,9-tetrahydro-5H-carbazole-l-carboxamide (0.10 equiv, 13 mmol) was added into the mixture in Reactor A, followed by the addition of 2-methyltetrahydrofuran (0.50 L/kg) and water (0.5 L/kg).

The overhead space of Reactor A was sparged with nitrogen at 1 mL/second for 40 min at 25 °C. The resulting mixture was then stirred at 70 °C for 3 h under a positive pressure of nitrogen (1.05 atm). The resulting mixture containing the activated catalyst was cooled to

25 °C and kept at 25 °C under a positive pressure of nitrogen before use.

To a 500 mL Chemglass reactor (Reactor B) were added water (6 L/kg) followed by K3PO4 (6 equiv., 813 mmol). The addition was exothermic. The mixture was stirred till the base was fully dissolved. The overhead space of Reactor B was sparged with nitrogen at 1 mL/second for 60 min at 25 °C. The K3PO4 solution in Reactor B was then kept under a positive pressure of nitrogen before use.

To Reactor A, which contained the activated catalyst, was added 4-bromo-3-fluoro-7-(l-hydroxy-l-methyl-ethyl)-6,7,8,94etrahydro-5H-carbazole-l-carboxarnide (0.90 equiv., 122 mmol), followed by THF (2.5 L/kg). Then (3-amino-2-methyl-phenyl)boronic acid hydrochloride (1.15 equiv., 156 mmol) and MeOH (2 L/kg) were added to Reactor A. The overhead space of Reactor A was sparged with nitrogen at 1 mL/second for 40 min. Then the reaction mixture in Reactor A was cooled to -10 °C under a positive pressure of nitrogen.

The K3PO4 aqueous solution in Reactor B was then transferred into Reactor A via a cannula while both reactors were kept under a positive pressure of N2. The rate of transfer was controlled so that the inner temperature in Reactor A was below 0 °C throughout the operation.

The resulting biphasic reaction mixture was stirred at 5 °C under a positive pressure of nitrogen. After 2.5 h at 5 °C, HPLC analysis of the reaction mixture showed

0.3 AP starting material remained. The reaction mixture was then warmed to 25 °C and stirred at 25 °C for 30 min. HPLC analysis of the reaction mixture showed 0.0 AP starting material remained.

N-acetyl-L-cysteine (1 kg/kg, 306 mmol) and water (2.5 L/kg) were added into Reactor A. The resulting mixture was stirred at 40 °C for 2 h then cooled to 25 °C. The bottom layer (aqueous layer) was discharged and the top layer (organic layer) was retained in the reactor.

Afterwards, THF (1 L/kg) and NaCl solution (13 mass%) in water (7 L/kg) were added into Reactor A, and the resulting mixture was stirred at 25 °C for lh. The bottom layer (aqueous layer) was discharged and the top layer (organic layer) was retained in the reactor.

The organic layer was filtered through a polyethylene filter. Then the reactor was rinsed with Me-THF (0.50 L/kg). The rinse was filtered through the polyethylene filter and combined with the filtrate. The solution was transferred into a clean 1 L reactor (Reactor C).

The mixture in Reactor C was concentrated under reduced pressure to 8.8 L/kg. (2 L/kg solvent was removed by distillation). At 50 °C, n-BuOH (4 L/kg) was added slowly over 2 h. The mixture was then stirred at 50 °C for 2.5 h, and a slurry was obtained.

The solvent was swapped to n-BuOH through constant volume distillation. During this operation, n-BuOH (8 L/kg) was used and 8 L/kg solvent was removed from Reactor C. The resulting mixture was stirred at 55 °C for 1 h and cooled to 25 °C over 1 h.

The slurry in Reactor C was filtered. The reactor rinsed with n-BuOH (2 L/kg).

The cake was then washed with this reactor rinse, followed by heptane (8 L/kg). The product was dried under vacuum at 55 °C for 24 h to afford (2S,5R)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide, which was isolated as an off-white solid powder (46.2 g, 86% yield).

HPLC analysis: (2S,5R)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide: 98.1 AP (19.2 min); (2S,5S)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide: 1.8 AP (19.9 min), (S)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide: 0.1 AP (20.9 min). Column: Waters XBridge BEH C18 S-2.5um 150 X 4.6mm; Solvent A: 10 mM sodium phosphate buffer pH 7; Solvent B: CH3CN:MeOH (50:50 v/v); Gradient: % B: 0 Min. 5%; 4 Min. 30%; 41 Min. 95%; 47 Min. 95%; Stop Time: 48 min; Flow Rate: 0.7 ml/min wavelength: 240 nm. ¾ NMR (500 MHz, DMSO-de) δ 10.76 (s, 1H), 8.09 (br s, 1H), 7.54 (d, J=10.7 Hz, 1H), 7.47 (br s, 1H), 6.96 (t, J=7.7 Hz, 1H), 6.72 (d, J=7.9 Hz, 1H), 6.41 (d, J=7.3 Hz, 1H), 4.90 (s, 2H), 4.19 (s, 1H), 2.91 (br dd, J=16.6, 4.0 Hz, 1H), 2.50-2.39 (m, 1H), 2.05-1.93 (m, 1H), 1.88-1.75 (m, 5H), 1.64-1.53 (m, 1H), 1.21-1.11 (m, 1H), 1.09 (s, 6H). 13C NMR (126 MHz, DMSO-de) δ 169.0 (d, J=2.7 Hz), 152.5 (d, J=229.8 Hz), 146.7, 139.1,

134.4, 132.0, 127.7 (d, J=4.5 Hz), 125.6, 123.3 (d, J=20.0 Hz), 120.5, 119.2, 1 15.1 (d, J=7.3 Hz), 1 14.3, 109.5(d, J=4.5 Hz), 107.2 (d, J=27.3 Hz), 70.9, 45.9, 27.6, 27.2, 25.3, 25.0, 22.7, 14.7.

Compound 7

propyl (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro- lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl)carbamate

N, N-Dimethylformamide (7.0 L, 7 L/kg) was charged into a reactor followed by the addition of (2S)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (1 kg, 2528 mmol, 1.0 eq.). 3-Fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid (0.774 kg, 3034 mmol, 1.2 eq.) was added to the reactor, followed by the addition of 1 -methylimidazole (0.267 kg, 3287 mmol, 1.3 eq) and methanesulfonic acid (0.122 kg, 1264 mmol, 0.5 eq.) at 20 °C. The reaction mixture was stirred for at 20 °C for 30 min to completely dissolve the reaction contents. The reaction mixture was cooled to 10 °C and EDAC (l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (0.679 kg, 3540 mmol, 1.4 eq) was charged into the reactor. An exotherm of approximately 4 °C was observed. The reaction mixture was stirred at 10 °C for 4 h.

After 4 hrs, the reaction mixture was warmed to 20 °C. Isopropyl acetate (25 L, 25 L/kg) was added to the reaction mixture followed by 25 wt% aqueous sodium chloride solution (2.5 L, 2.5 L/kg) and 1.0 M aqueous hydrochloric acid (2.5 L, 2.5 L/kg). The reaction mixture was stirred for 30 min. The agitation was stopped and the bottom aqueous layer was separated. Water (5 L, 5 L/kg) was charged to the rich organic solution and stirred for 30 min. The agitation was stopped and the bottom aqueous layer was separated. Next, 2.5% aqueous sodium bicarbonate solution (10 L, 10 L/kg) was charged to the rich organic solution and stirred for 30 min. The agitation was stopped and the bottom aqueous layer was separated. Water (10 L, 10 L/kg) was charged to the rich organic solution and stirred for 30 min. The agitation was stopped and the bottom aqueous layer was separated. The rich organic solution was concentrated under reduced pressure (90 mbar and 40 °C jacket temperature) to 7 L/kg volume. Dichloromethane (5 L, 5 L/kg) was charged to the product rich isopropyl acetate solution at 20 °C. Seeds of propyl (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl)carbamate (10 g, 1%) were charged and a thin slurry formed. Heptane (7 L, 7 L/kg) was charged to the above slurry slowly over 1 hr at 25 °C and stirred for another 1 h before cooling 20 °C over 30 min. The resultant slurry was stirred for 4-6 hrs at 20 °C. The slurry was filtered over a laboratory Buchner funnel. The wet cake was washed with a dichloromethane-heptane mixture (10:7 ratio, 12 vol). The wet cake was dried in a vacuum oven at 25 mm Hg vacuum and 50 °C until the residual heptane was <13 wt% in the solid to provide 1.5 kg of propyl (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl) carbamate in 94% yield. The product was a mixture of four amide rotational isomers. ¾ NMR (400 MHz, DMSO-de) δ 10.79 (br s, 1H), 9.96 (m, 1H), 8.07 (br s, 1H), 7.50 (m, 6H), 7.29 (m, 1H), 7.09 (m, 1H), 4.15 (m, 1H), 3.89 (m, 2H), 3.19 (br s, 1H), 3.13 (br s, 2H), 2.90 (m, 1H), 2.44 (m, 1H), 1.97 (m, 3H), 1.82 (m, 3H), 1.50 (m, 3H), 1.26 (m, 5H), 1.09 (m, 7H), 0.85 (m, 4H), 0.70 (m, 2H). 13C NMR (101 MHz, DMSO-de) δ 168.33, 168.32, 164.85, 164.55, 159.38, 159.16, 156.93, 156.69, 154.90, 154.74, 153.14, 150.86, 139, 15, 139.11, 137.96, 137.89, 137.36, 137.23, 135.75, 135.68, 135.64, 134.77, 134.68, 132.57, 132.51, 132.46, 132.42, 131.50, 128.98 (m), 128.26 (m), 127.05, 127.01, 125.99, 125,76, 124.97, 124.83, 124.06, 121.48, 121.40, 121.28, 121.20, 117.90, 117.86, 117.70, 117.65, 115.19, 115.15, 115.12, 115.07, 108.69, 108.65, 106.87, 106.60, 70.39, 66.83, 66.80, 66.73, 45.32, 37.38, 37.15, 31.23, 28.35, 27.05, 26.68, 24.85, 24.61, 22.27, 22.07, 21.84, 21.75, 14.98, 14.93, 14.86, 14.84, 13.87, 10.11, 9.89.

HPLC Analysis: Column: Zorbax Eclipse Plus C18 3.5 um, 150 x 4.6 mm ID;

Solvent A: 10 mM ammonium formate in water-MeOH (90: 10); Solvent B: C¾CN :

MeOH (30:70 v/v); Gradient: % B: 0 Min. 50%; 25 Min. 81 %; 26 Min. 100%; 30 Min. 100%; Stop Time: 30 min; Flow Rate: 1 ml/min; Wavelength: 240 nm. The retention time of propyl (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl) carbamate wasl4.6 min. The retention time of 3-fluoro-2-(methyl(propoxycarbonyl) amino)benzoic acid was 2.6 min. The retention time of (2S)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide was 6.1 min.

Compound 8

6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l ,2-dihydroquinazolin-3(4H)-yl)-2- methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide


(8)

To a 1 L round bottom flask with stir bar was added propyl (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl)carbamate (100 g, 148 mmol, 93.5 mass%) followed by MeTHF (500 mL, 4990 mmol, 100 mass%). The mixture was stirred at room temperature for 10 minutes to ensure complete dissolution. Next, 150 mL of MeTHF was added, and an azeotropic distillation to remove water was performed at 50 °C and 70 torr. The KF was measured to be 424 ppm. This solution is termed the “Compound 8 solution.”

To a 2 L Chemglass reactor was charged MeTHF (2000 mL, 19900 mmol, 100 mass%) followed by lithium fert-butoxide (7.9 mL, 7.9 mmol, 1 mol/L). The KF of MeTHF was measured to be 622 ppm. The Compound 8 solution was added dropwise

over 2 hours at room temperature via a Simdos pump. After the addition was complete, the reaction mixture was maintained at temperature for 15 minute.

MeOH (200 mL, 4940 mmol, 100 mass%) was then added to the reactor followed by the addition of acetic acid (0.5 mL, 9 mmol, 100 mass%). The reaction mixture was distilled to 5 volumes of organics (60 mbar pressure, jacket temperature = 40 °C). After the distillation, acetone (150 mL, 2000 mmol, 100 mass%) was added to the thick slurry as the solution warmed to 35 °C. Once at 35 °C, MeOH (550 mL, 13600 mmol, 100 mass%) was charged to the reactor, re-dissolving the batch to provide a yellow solution. The reaction mixture was cooled over 1 hour to 20 °C resulting in crystallization of the product. Ten heat cycles were performed. Starting at 20 °C, the batch was heated to 35 °C over 45 minutes, held at 35 °C for 10 minutes, cooled 20 °C over 60 minutes, and held at 20 °C for 10 minutes. After the heat cycles, the slurry was maintained at room temperature for 1 hour at room temperature. Heptane (1100 mL, 7510 mmol, 100 mass%) was added over 4 hours at 20 °C with agitation via a Simdos pump. After the addition, the slurry aged to 20 °C overnight. The product was isolated by vacuum filtration and washed twice with MeOH (200 mL, 4940 mmol, 100 mass%). The product was dried on a filter with vacuum for 1.5 h to afford 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide at 89.4% corrected yield (80.52g, 6 wt % MeOH, Purity by HPLC: 99.32 AP; Retention time (11.65 min)).

¾ NMR (500MHz, DMSO-de) 10.78 (s, 1H), 8.07 (br. s., 1H), 7.95 (d, J=7.8 Hz, 1H), 7.72 (dd, J=14.2, 8.0 Hz, 1H), 7.56 (d, J=10.8 Hz, 1H), 7.45 (br. s., 1H), 7.42-7.36 (m, 1H), 7.34 (d, J=6.9 Hz, 1H), 7.34-7.31 (m, 1H), 7.29 (dd, J=7.5, 1.3 Hz, 1H), 4.17 (s, 1H), 3.73 (d, J=8.0 Hz, 3H), 2.91 (dd, J=16.8, 4.4 Hz, 1H), 2.48-2.37 (m, 1H), 1.98-1.89 (m, 2H), 1.87 (d, J=11.0 Hz, 1H), 1.76 (s, 3H), 1.59 (td, J=l 1.5, 4.1 Hz, 1H), 1.20-1.12 (m, 1H), 1.11 (s, 6H).

13C NMR (126MHz, DMSO-de) 168.2 (d, J=1.8 Hz, 1C), 160.1 (d, J=3.6 Hz, 1C), 151.9 (d, J=228.9 Hz, 1C), 150.5 (d, J=41.8 Hz, 1C), 148.7 (d, J=205.3 Hz, 1C), 139.2, 135.1, 135.0, 134.8, 131.4, 130.6, 130.0 (d, J=7.3 Hz, 1C), 128.5, 127.1 (d, J=4.5 Hz, 1C), 125.7, 124.3 (d, J=2.7 Hz, 1C), 123.6 (d, J=8.2 Hz, 1C), 123.0 (d, J=23.6 Hz, 1C), 120.8 (d, J=20.0 Hz, 1C), 118.4, 115.3 (d, J=7.3 Hz, 1C), 108.8 (d, J=5.4 Hz, 1C), 106.7 (d, J=28.2 Hz, 1C), 70.4, 45.4, 34.3 (d, J=14.5 Hz, 1C), 27.1, 26.8, 24.8, 24.7, 22.1, 14.5.

HPLC Analysis: Column: Chiralcel OX-3R 3um 4.6 x 150 mm; Oven

Temperature: 50 °C; Solvent A: 0.05%TFA Water/ ACN (95:5); Solvent B: 0.05%TFA Water/ ACN (5:95); Gradient % B: 0 Min. 0%; 7 Min. 55%; 11 Min. 55%; 14 Min. 100%; Stop Time: 17 Min.; Flow Rate: 1.5 ml/min; wavelength: 225 nm. (2-((3-((2S)-8-carbamoyl-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazol-5-yl)-2-methylphenyl)carbamoyl)-6-fluorophenyl)(methyl)carbamate: 0.00 AP (9.85 min).

Alternative Preparation of Compound 8

To a 2.5 L Chemglass reactor with agitator were added 2-Me-THF (162.4 g, 1885 mmol, 100 mass%, 189 mL, 11.83) and DMF (179.5 g, 2456 mmol, 100 mass%, 190 mL, 15.41), followed by the addition of (2S)-5-(3-amino-2-methylphenyl)-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (63.03 g, 63.03 mL, 159.4 mmol, 63.03 g), 3-fluoro-2-(methyl(propoxycarbonyl)amino)benzoic acid (44.77 g, 44.77 mL, 175.4 mmol, 44.77 g), and 1 -Me-Imidazole (16.99 g, 16.48 mL, 206.9 mmol, 16.99 g). With agitation, MSA (7.66 g, 5.23 mL, 79.7 mmol, 7.66 g) was added at -20 °C, and a slight exotherm to 26 °C was observed. The reaction mixture was cooled to 10 °C and ED AC (42.73 g, 42.73 mL, 222.9 mmol, 42.73 g) was added as a solid followed by a DMF rinse (60.4 g, 63.9 mL, 826 mmol, 60.4 g). The reaction mixture was aged overnight at 10 °C with agitation. An aliquot was taken and subjected to HPLC analysis to confirm reaction completion.

The batch temperature was increased to 15 °C, and 2-Me-THF (923.96 g, 10727 mmol, 100 mass%, 1080 mL, 67.31) was charged to the reactor, followed by a saturated aqueous brine solution (158 mL, 835.8 mmol, 26 mass%, 158 mL, 5.244) and an aqueous 2.0 M HCl solution (78 mL, 78 mmol, 1.0 mol/L, 78 mL, 0.49). The batch temperature was then increased to 20 °C. The biphasic mixture was agitated for 15 min and allowed to settle for 5 min. An saturated aqueous brine solution (157 mL, 830.5 mmol, 26 mass%, 157 mL, 5.211) and an aqueous 2.0 M HCl solution (78 mL, 78 mmol, 1.0 mol/L, 78 mL, 0.49) were then added to the reactor. The biphasic mixture was agitated for 15 min, allowed to settle for 5 min, and the aqueous layer was removed. Water (634.6 g, 35230 mmol, 100 mass%, 634.6 mL, 221.0) was then added to the reactor. The biphasic mixture was agitated for 15 min, allowed to settle for 5 min, and the aqueous layer was removed. Next, 10 w/w% aqueous NaHCC solution (164.2 g, 97.73 mmol, 5 mass%,

158.2 mL, 0.6132) and water (476.3 g, 26440 mmol, 100 mass%, 476.3 mL, 165.9) were added to the reactor. The biphasic mixture was agitated for 15 min, settled for 5 min, and the aqueous layer was removed. A saturated aqueous brine solution (752.9 g, 3349 mmol, 26 mass%, 633.2 mL, 21.02) was then added to the reactor. The biphasic mixture was agitated for 30 min, allowed to settle for 5 min, and the aqueous layer was removed.

The organic stream was distilled to 6 volumes (380 mL) at a pressure of 200 mbar, a jacket temperature of 60 °C, and a batch temperature of -35 °C. 2-Me-THF (765 g, 8881.6 mmol, 100 mass%, 891 mL, 55.73) was charged to the reactor. The organic solution was distilled to 6 volumes (380 mL) at a pressure of 200 mbar, a jacket temperature of 60 °C, and a batch temperature of -35 °C. 2-Me-THF (268.5 g, 3117 mmol, 100 mass%, 313 mL, 19.56) was charged to the reactor. The organic solution was distilled to 6 volumes (380 mL) at a pressure of 200 mbar, a jacket temperature of 60 °C, and a batch temperature of -35 °C. The concentrated stream was polish filtered through a 0.4 μιη PTFE filter. The reactor was rinsed with 2-Me-THF (134.6 g, 1563 mmol, 100 mass%, 157 mL, 9.806) and the rinse was passed through the PTFE filter. This solution was termed “organic solution.”

To a clean, dry, 2.5 L Chemglass reactor were added LiOtBu 1.0 M in THF (9.91 g, 11.2 mmol, 1 mol/L, 11.2 mL, 0.0700) and 2-Me-THF (1633.3 g, 18963 mmol, 100 mass%, 1900 mL, 119.0). The organic solution was charged to the reactor, with agitation, over 2 hours (at a rate of -100 mL/h) via a sim-dos pump. The reaction mixture was aged 10 minutes upon completion of the addition. An aliquot was taken and subjected to HPLC analysis to confirm reaction completion.

Acetic acid (1.03 g, 17.2 mmol, 100 mass%, 0.983 mL, 0.108) and methanol (150 g, 4681.41 mmol, 100 mass%, 189 mL, 29.37) were charged to the reactor. The organic stream was distilled to 16.5 vol Me-THF. Acetone (638.4 g, 10990 mmol, 100 mass%, 810 mL, 68.97) was added to the reactor and the organic stream was distilled to 9 vol at a pressure of 100 mbar and ajacket temperatures of less than 40 °C. The organic stream was heated to 35 °C, and methanol (400 g, 12483.8 mmol, 100 mass%, 505 mL, 78.33) was added. The stream was cooled to 20 °C to induce crystallization.

Heat cycles were performed for -15 h by heating the batch to 35 °C over 20 min, holding for 10 min, cooling to 20 °C over 20 min, and holding 10 min. After the heat cycles, heptane (686 g, 6846.10 mmol, 100 mass%, 1000 mL, 42.96) was added over 4 hours via a sim-dos pump. The slurry was aged for 2 h. The product was filtered, washed with methanol (152.2 g, 4750 mmol, 100 mass%, 192 mL, 29.81) to afford 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l -methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (68.4 g, 1 19 mmol, 100 mass%, 75.0% Yield, 68.4 mL, 0.750).

Comparative Process Disclosed in US 9,334,290

Intermediates 25 and 26

(R)-5-Bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- carboxamide (1-25), and

(S)-5-Bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8- -26)

A sample of racemic 5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide [Intermediate 24] was separated by chiral supercritical fluid chromatography as follows: column: CHIRALPAK® OD-H (3 x 25 cm, 5μηι); Mobile Phase: CC -MeOH (70:30) at 150 mL/min, 40 °C. The first peak eluting from the column provided (R)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide [Intermediate 25]. The second peak eluting from the column provided (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide [Intermediate 26]. The mass spectra and ¾ NMR spectra of the two enantiomers were the same. Mass spectrum m/z 369, 371 (M+H)+. ¾ NMR (500 MHz, DMSO-de) δ 10.96 (s, 1H), 8.07 (br. s., 1H), 7.55 (d, J=10.3 Hz, 1H), 7.50 (br. s., 1H), 4.24 (s, 1H), 3.26 (dd, J=15.8, 4.4 Hz, 1H), 2.93 (dd, J=17.1, 4.6 Hz, 1H), 2.72 (t, J=11.7 Hz, 1H), 2.48-2.40 (m, 1H), 2.12 (d, J=9.2 Hz, 1H), 1.70-1.62 (m, 1H), and 1.32 (qd, J=12.4, 5.3 Hz, 1H).

Alternative SFC Separation to Give Intermediate 26:

CHIRALPAK® AD-H (3 x 25 cm, 5 μηι); Mobile Phase: C02-MeOH (55:45) at

150 mL/min, 40 °C. The first peak eluting from the column provided (S)-5-bromo-6- fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxarnide

[Intermediate 26]. The second peak eluting from the column provided (R)-5-bromo-6- fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxarnide

[Intermediate 25].

Example 28

6-Fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2- methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-

Following the procedure used to prepare Example 27, (S)-5-bromo-6-fluoro-2-(2- hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (single enantiomer) [Intermediate 26] (0.045 g, 0.122 mmol) and 8-fluoro-l-methyl-3-(S)-(2-methyl-3- (4,4,5, 5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)quinazoline-2,4(lH,3H)-dione

[Intermediate 10] (0.065 g, 0.158 mmol) were converted into 6-fluoro-5-(3-(S)-(8-fluoro- l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2- hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide (mixture of two atropisomers) as a yellow solid (0.035 g, 49% yield). Separation of a sample of this material by chiral super-critical fluid chromatography, using the conditions used to separate Example 27, provided (as the first peak to elute from the column) 6-fluoro-5-(R)-(3-(S)-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxarnide. The chiral purity was determined to be greater than 99.5%. The relative and absolute configurations were determined by x-ray crystallography. Mass spectrum m/z 573 (M+H)+. ¾ NMR (500 MHz, DMSO-de) δ 10.77 (s, 1H), 8.05 (br. s., 1H), 7.94 (dd, J=7.9, 1.2 Hz, 1H), 7.56-7.52 (m, 1H), 7.43 (br. s., 1H), 7.40-7.36 (m, 1H), 7.35-7.30 (m, 2H), 7.28 (dd, J=7.5, 1.4 Hz, 1H), 4.15 (s, 1H), 3.75-3.70 (m, 3H), 2.90 (dd, J=16.8, 4.6 Hz, 1H), 2.47-2.39 (m, 1H), 1.93-1.82 (m, 3H), 1.74 (s, 3H), 1.57 (td, J=l 1.7, 4.2 Hz, 1H), 1.16-1.11 (m, 1H), and 1.10 (d, J=1.9 Hz, 6H). [a]D: +63.8° (c 2.1, CHCh). DSC melting point onset temperature = 202.9 °C (heating rate = 10 °C/min.).

Alternative Synthesis of Example 28:

A mixture of (S)-5-bromo-6-fluoro-2-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide [Intermediate 26] (5.00 g, 13.54 mmol), 8-fluoro-l-methyl-3-(S)-(2-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl)quinazoline-2,4(lH,3H)-dione [Intermediate 10] (6.67 g, 16.25 mmol), tripotassium phosphate (2 M in water) (20.31 mL, 40.6 mmol), and tetrahydrofuran (25 mL) was subjected to 3 evacuate-fill cycles with nitrogen. The mixture was treated with l,l’-bis(di-fert-butylphosphino)ferrocene palladium dichloride (0.441 g, 0.677 mmol) and the mixture was subjected to 2 more evacuate-fill cycles with nitrogen. The mixture was stirred at room temperature overnight, then was diluted with EtOAc, washed sequentially with water and brine, and dried and concentrated. The residue was purified by column chromatography on silica gel, eluting with EtOAc-hexanes (sequentially 50%, 62%, 75% and 85%), to provide 6-fluoro-5-(3-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3-(S)-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide as a white solid (6.58 g, 85% yield).

Material prepared by this method (40.03 g, 69.9 mmol) was separated by chiral super-critical fluid chromatography to give (2S, 5R)-6-fluoro-5-(3-(8-fluoro-l-methyl-2,4-dioxo-l,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-lH-carbazole-8-carboxamide. Further purification was achieved

by suspending this material in methanol, sonicating for 5 min, collection of the solid by filtration, rinsing the collected solid with methanol and drying at room temperature under reduced pressure to give a white solid (22.0 g, 90% yield).

REFERENCES

1: Watterson SH, De Lucca GV, Shi Q, Langevine CM, Liu Q, Batt DG, Beaudoin Bertrand M, Gong H, Dai J, Yip S, Li P, Sun D, Wu DR, Wang C, Zhang Y, Traeger SC, Pattoli MA, Skala S, Cheng L, Obermeier MT, Vickery R, Discenza LN, D’Arienzo CJ, Zhang Y, Heimrich E, Gillooly KM, Taylor TL, Pulicicchio C, McIntyre KW, Galella MA, Tebben AJ, Muckelbauer JK, Chang C, Rampulla R, Mathur A, Salter-Cid L, Barrish JC, Carter PH, Fura A, Burke JR, Tino JA. Discovery of 6-Fluoro-5-(R)-(3-(S)-(8-fluoro-1-methyl-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl )-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (BMS-986142): A Reversible Inhibitor of Bruton’s Tyrosine Kinase (BTK) Conformationally Constrained by Two Locked Atropisomers. J Med Chem. 2016 Oct 13;59(19):9173-9200. PubMed PMID: 27583770.

(a) Watterson, S. H.De Lucca, G. V.Shi, Q.Langevine, C. M.Liu, Q.Batt, D. G.Bertrand, M. B.Gong, H.Dai, J.Yip, S.Li, P.Sun, D.Wu, D.-R.Wang, C.Zhang, Y.Traeger, S. C.Pattoli, M. A.Skala, S.Cheng, L.Obermeier, M. T.Vickery, R.Discenza, L. N.D’Arienzo, C. J.Zhang, Y.Heimrich, E.Gillooly, K. M.Taylor, T. L.Pulicicchio, C.McIntyre, K. W.Galella, M. A.Tebben, A. J.Muckelbauer, J. K.Chang, C.Rampulla, R.Mathur, A.Salter-Cid, L.Barrish, J. C.Carter, P. H.Fura, A.Burke, J. R.Tino, J. A. Discovery of 6-Fluoro-5-(R)-(3-(S)-(8-fluoro-1-methyl-2,4-dioxo-1,2-dihydroquinazolin-3(4H)-yl)-2-methylphenyl)-2-(S)-(2-hydroxypropan-2-yl)-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (BMS-986142): A Reversible Inhibitor of Bruton’s Tyrosine Kinase (BTK) Conformationally Constrained by Two Locked AtropisomersJ. Med. Chem. 2016599173DOI: 10.1021/acs.jmedchem.6b01088
(b) De Lucca, G. V.Shi, Q.Liu, Q.Batt, D. G.Bertrand, M. B.Rampulla, R.Mathur, A.Discenza, L.D’Arienzo, C.Dai, J.Obermeier, M.Vickery, R.Zhang, Y.Yang, Z.Marathe, P.Tebben, A. J.Muckelbauer, J. K.Chang, C. J.Zhang, H.Gillooly, K.Taylor, T.Pattoli, M. A.Skala, S.Kukral, D. W.McIntyre, K. W.Salter-Cid, L.Fura, A.Burke, J. R.Barrish, J. C.Carter, P. H.Tino, J. A. Small Molecule Reversible Inhibitors of Bruton’s Tyrosine Kinase (BTK): Structure–Activity Relationships Leading to the Identification of 7-(2-Hydroxypropan-2-yl)-4-[2-methyl-3-(4-oxo-3,4-dihydroquinazolin-3-yl)phenyl]-9H-carbazole-1-carboxamide (BMS-935177)J. Med. Chem. 2016597915DOI: 10.1021/acs.jmedchem.6b00722
Watterson, S.H.; De Lucca, G.V.; Shi, Q.; et al.
Twisted road to the discovery of BMS-986142: Using conformationally locked atropisomers to drive potency in a reversible inhibitor of Brutonas tyrosine kinase (BTK)
255th Am Chem Soc (ACS) Natl Meet (March 18-22, New Orleans) 2018, Abst MEDI 6

////////////BMS-986142, BMS 986142, BMS986142,  phase II,  clinical development,  Bristol-Myers Squibb, rheumatoid arthritis, primary Sjogren’s syndrome,

CN1C(=O)N(C(=O)c2cccc(F)c12)c3cccc(c3C)c4c(F)cc(C(=O)N)c5[nH]c6C[C@H](CCc6c45)C(C)(C)O

FDA approves first treatment Libtayo (cemiplimab-rwlc) for advanced form of the second most common skin cancer


FDA approves first treatment for advanced form of the second most common skin cancer

New drug targets PD-1 pathway

The U.S. Food and Drug Administration today approved Libtayo (cemiplimab-rwlc) injection for intravenous use for the treatment of patients with metastatic cutaneous squamous cell carcinoma (CSCC) or locally advanced CSCC who are not candidates for curative surgery or curative radiation. This is the first FDA approval of a drug specifically for advanced CSCC.

September 28, 2018

Release

The U.S. Food and Drug Administration today approved Libtayo (cemiplimab-rwlc) injection for intravenous use for the treatment of patients with metastatic cutaneous squamous cell carcinoma (CSCC) or locally advanced CSCC who are not candidates for curative surgery or curative radiation. This is the first FDA approval of a drug specifically for advanced CSCC.

Libtayo works by targeting the cellular pathway known as PD-1 (protein found on the body’s immune cells and some cancer cells). By blocking this pathway, the drug may help the body’s immune system fight the cancer cells.

“We’re continuing to see a shift in oncology toward identifying and developing drugs aimed at a specific molecular target. With the Libtayo approval, the FDA has approved six immune checkpoint inhibitors targeting the the PD-1 / PD-L1 pathway for treating a variety of tumors, from bladder to head and neck cancer, and now advanced CSCC,” said Richard Pazdur, M.D., director of the FDA’s Oncology Center of Excellence and acting director of the Office of Hematology and Oncology Products in the FDA’s Center for Drug Evaluation and Research. “This type of cancer can be difficult to treat effectively when it is advanced and it is important that we continue to bring new treatment options to patients.”

CSCC is the second most common human cancer in the United States with an estimated annual incidence of approximately 700,000 cases. The most common form of skin cancer is basal cell cancer. Squamous cells are thin, flat cells that look like fish scales and are found in the tissue that forms the surface of the skin. CSCC usually develops in skin areas that have been regularly exposed to the sun or other forms of ultraviolet radiation. While the majority of patients with CSCC are cured with surgical resection, a small percentage of patients will develop advanced disease that no longer responds to local treatments including surgery and radiation. Advanced CSCC may cause disfigurement at the site of the tumor and local complications such as bleeding or infection, or it may spread (metastasize) to local lymph nodes, distant tissues and organs and become life-threatening.

The safety and efficacy of Libtayo was studied in two open label clinical trials. A total of 108 patients (75 with metastatic disease and 33 with locally-advanced disease) were included in the efficacy evaluation. The study’s primary endpoint was objective response rate, or the percentage of patients who experienced partial shrinkage or complete disappearance of their tumor(s) after treatment. Results showed that 47.2 percent of all patients treated with Libtayo had their tumors shrink or disappear. The majority of these patients had ongoing responses at the time of data analysis.

Common side effects of Libtayo include fatigue, rash and diarrhea. Libtayo must be dispensed with a patient Medication Guide that describes uses of the drug and its serious warnings. Libtayo can cause the immune system to attack normal organs and tissues in any area of the body and can affect the way they work. These reactions can sometimes become severe or life-threatening and can lead to death. These reactions include the risk of immune-mediated adverse reactions including lung problems (pneumonitis), intestinal problems (colitis), liver problems (hepatitis), hormone gland problems (endocrinopathies), skin (dermatologic) problems and kidney problems. Patients should also be monitored for infusion-related reactions.

Libtayo can cause harm to a developing fetus; women should be advised of the potential risk to the fetus and to use effective contraception.

The FDA granted this application Breakthrough Therapy and Priority Reviewdesignations.

The FDA granted the approval of Libtayo to Regeneron Pharmaceuticals, Inc.

////////////Libtayo, cemiplimab-rwlc, FDA 2018,  Breakthrough Therapy,  Priority Review

Icosapent ethyl, イコサペント酸エチル


DB08887.png

Ethyl eicosapentaenoate.png

Icosapent ethyl

330.5042 , C22H34O2

cas 86227-47-6 / 73310-10-8

ethyl (5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenoate

Ethyl eicosapentaenoic acid

イコサペント酸エチル

(5Z,8Z,11Z,14Z,17Z)-Eicosapetaenoic acid ethyl ester
(all-Z)-5,8,11,14,17-Eicosapentaenoic acid ethyl ester
5,8,11,14,17-Eicosapentaenoic acid, ethyl ester, (5Z,8Z,11Z,14Z,17Z)- [ACD/Index Name]
5,8,11,14,17-Eicosapentaenoic acid, ethyl ester, (all-Z)-
6GC8A4PAYH
86227-47-6 [RN]
all-cis-5,8,11,14,17-Eicosapentaenoic Acid Ethyl Ester
Timnodonic acid ethyl ester
Vascepa
  • 5,8,11,14,17-Eicosapentaenoic acid, ethyl ester, (all-Z)-
  • (5Z,8Z,11Z,14Z,17Z)-5,8,11,14,17-Eicosapentaenoic acid ethyl ester
  • (all-Z)-5,8,11,14,17-Eicosapentaenoic acid ethyl ester
  • AMR 101
  • C20:5 n-3 Ethyl ester
  • Epadel
  • Epadel S 300
  • Ethyl (5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoate
  • Ethyl all-Z-5,8,11,14,17-eicosapentanenoate
  • Ethyl all-cis-5,8,11,14,17-eicosapentaenoate
  • Ethyl eicosapentaenoate
  • Ethyl icosapentate
  • Icosapent ethyl
  • Incromega EPA
  • Timnodonic acid ethyl ester
  • Vascepa
  • cis-Eicosapentaenoic acid ethyl ester

(all-Z)-5,8,11,14,17-Eicosapentaenoic acid ethyl ester; Ethyl all-cis-5,8,11,14,17-eicosapentaenoate;Timnodonic acid ethyl ester; cis-Eicosapentaenoic acid ethyl ester; Ethyl (5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoate; Epadel; Icosapent; EPA ethyl ester; E-EPA; Ethyl eicosapentaenoate; OMEGA-3 ACIDS ETHYL ESTER; EPA-E;

AMARIN PHARMACEUTICALS IRELAND LTD

AMR 101 / AMR-101 / AMR101

Icosapent ethyl or ethyl eicosapentaenoic acid is a synthetic derivative of the omega-3 fatty acid eicosapentaenoic acid (EPA). It is used as adjunct therapy for severe hypertriglyceridemia (TG levels > 500 mg/dL). FDA approved on July 26, 2012.

In 2000, Amarin licensed exclusive U.S. rights to icosapent ethyl ester from the Scottish company Laxdale, and acquired the company in July 2004. In 2015, the product was licensed to Eddingpharm by Amarin for the development and commercialization in China, Hong Kong and Taiwan. Fast-track status has been granted in the U.S. for the treatment of HD. Orphan drug designation was assigned to the compound for this indication in both the U.S. and E.U.

fda

IND 107616 was submitted on 25 March 2010 for the indication of severe hypertriglyceridemia; Epanova had been previously investigated for the treatment of Crohn’s Disease under IND in the Division of Gastroenterology Products. An end-of-phase 2 (EOP2) meeting was held on 02 June 2010. Regarding the indication under consideration at this time, a special protocol assessment (SPA) for the single phase 3 trial OM-EPA-003 (also known as “EVOLVE”) was submitted 02 July 2010 and ultimately agreed upon, after amendments, on 22 October 2010. On 25 April 2012, the applicant proposed an alternative to conducting a thorough QTc study by assessing ECGs recorded during OM-EPA-003; this was found acceptable. A clinical pre-NDA meeting was held on 14 November 2012. The nonclinical development strategy was found reasonable. A clinical package containing OM-EPA-003 (pivotal) and OMEPA-004 (a 6-week phase 3 trial , with long-term safety supported by data from the former Crohn’s disease program (“EPIC” trials), was found adequate for submission. Agreement was reached regarding the clinical pharmacology portion of the submission. Details regarding data pooling for the Integrated Summary of Safety (ISS) were found acceptable

from the former Crohn’s disease program (“EPIC” trials), was found adequate for submission. Agreement was reached regarding the clinical pharmacology portion of the submission. Details regarding data pooling for the Integrated Summary of Safety (ISS) were found acceptable

CMC Drug Substance & Drug Product Chemistry, manufacturing, and controls data related to both the drug substance (omega-3- carboxylic acids) and drug product (Epanova Capsules 1 g) are detailed in the review by Martin Haber, PhD, and Xavier Ysern, PhD. They recommend the NDA for approval. There are no pending CMC issues. The drug substance at sites in Nova Scotia and Prince Edward Island, Canada, from crude fish oil obtained from fish It is a complex mixture of PUFAs, predominantly the omega-3 acids EPA (55%), DHA (20%), and docosapentaenoic acid %). It consistently contains omega-3 and omega-6 PUFA components: total omega-3 fatty acids are limited to not less than % and total omega-6 fatty acids are limited to not more than %. The drug substance also contains 0.3% (m/m) α-tocopherol as . During purification, . Environmental pollutants (heavy metals, pesticides, are controlled by specific tests on the drug substance . Drug substance specifications include tests for acid value, saponification value, ester value, peroxide value, p-anisidine value, total oxidation value, cholesterol, oligomers, , fatty acid composition (PUFAs, EPA, DHA, DPA, total omega-3 fatty acids, total omega-6 fatty acids, other polyunsaturated fatty acids, As described in the review by Drs. Haber and Ysern, the qualitative identify of the drug substance was developed by examining consistencies of peak patterns across 21 discrete lots: there are omega-3 and omega-6 PUFA peaks consistently present in the GC chromatograms (although not necessarily always above the limit of quantitation), which can be used to establish the fingerprint identity of omega-3-carboxylic acids . The quantitative fatty acid composition is given in the table below, excerpted from p. 25 of their review:

Ethyl eicosapentaenoic acid (E-EPAicosapent ethyl) is a derivative of the omega-3 fatty acid eicosapentaenoic acid (EPA) that is used in combination with changes in diet to lower triglyceride levels in adults with severe (≥ 500 mg/dL) hypertriglyceridemia. This was the second class of fish oil-based drug to be approved for use as a drug and was approved by the FDA in 2012. These fish oil drugs are similar to fish oil dietary supplements but the ingredients are better controlled and have been tested in clinical trials.

The company that developed this drug, Amarin Corporation, challenged the FDA’s ability to limit its ability to market the drug for off-label use and won its case on appeal in 2012, changing the way the FDA regulates pharmaceutical marketing.

Medical use

E-EPA is used in addition to changes in diet to reduce triglyceride levels in adults with severe (≥ 500 mg/dL) hypertriglyceridemia.[1]

Intake of large doses (2.0 to 4.0 g/day) of long-chain omega-3 fatty acids as prescription drugs or dietary supplements are generally required to achieve significant (> 15%) lowering of triglycerides, and at those doses the effects can be significant (from 20% to 35% and even up to 45% in individuals with levels greater that 500 mg/dL). It appears that both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) lower triglycerides, however, DHA alone appears to raise low-density lipoprotein (the variant which drives atherosclerosis; sometimes very inaccurately called: “bad cholesterol”) and LDL-C values (always only a calculated estimate; not measured by labs from person’s blood sample for technical and cost reasons), whilst EPA alone, does not and instead lowers the parameters aforementioned.[2]

Other fish-oil based drugs

There are other omega-3 fish oil based drugs on the market that have similar uses and mechanisms of action.[3]

Dietary supplements

There are many fish oil dietary supplements on the market.[8] There appears to be little difference in effect between dietary supplements and prescription forms of omega-3 fatty acids, but EPA and DHA ethyl esters (prescription forms) work less well when taken on an empty stomach or with a low-fat meal.[2] The ingredients of dietary supplements are not as carefully controlled as prescription products and have not been fixed and tested in clinical trials, as prescription drugs have,[9] and the prescription forms are more concentrated, requiring fewer capsules to be taken and increasing the likelihood of compliance.[8]

Side effects

Special caution should be taken with people who have with fish and shellfish allergies.[1] In addition, as with other omega-3 fatty acids, taking E-EPA puts people who are on anticoagulants at risk for prolonged bleeding time.[1][2] The most commonly reported side effect in clinical trials has been joint pain; some people also reported pain in their mouth or throat.[1] E-EPA has not been tested in pregnant women is rated pregnancy category C; it is excreted in breast milk and the effects on infants are not known.[1]

Pharmacology

After ingestion, E-EPA is metabolized to EPA. EPA is absorbed in the small intestine and enters circulation. Peak plasma concentration occurs about 5 hours after ingestion and the half-life is about 89 hours. EPA is metabolized mostly in the liver like other dietary fatty acids.[1]

Mechanism of action

EPA, the active metabolite of E-EPA, like other omega-3 fatty acid based drugs, appears to reduce production of triglycerides in the liver, and to enhance clearance of triglycerides from circulating very low-density lipoprotein (VLDL) particles; the way it does that is not clear, but potential mechanisms include increased breakdown of fatty acids; inhibition of diglyceride acyltransferase which is involved in biosynthesis of triglycerides in the liver; and increased activity of lipoprotein lipase in blood.[1][3]

Physical and chemical properties[edit]

E-EPA is an ethyl ester of eicosapentaenoic acid, which is an omega-3 fatty acid.[1]

History

In July 2012, the US Food and Drug Administration approved E-EPA for severe hypertriglyceridemia as an adjunct to dietary measures; Amarin Corporation had developed the drug.[10]

E-EPA was the second fish-oil drug to be approved, after omega-3 acid ethyl esters (GlaxoSmithKline‘s Lovaza which was approved in 2004[11]) and sales were not as robust as Amarin had hoped. The labels for the two drugs were similar, but doctors prescribed Lovaza for people who had triglycerides lower than 500 mg/dL based on some clinical evidence. Amarin wanted to actively market E-EPA for that population as well which would have greatly expanded its revenue, and applied to the FDA for permission to do so in 2013, which the FDA denied.[12] In response, in May 2015 Amarin sued the FDA for infringing its First Amendment rights,[13] and in August 2015 a judge ruled that the FDA could not “prohibit the truthful promotion of a drug for unapproved uses because doing so would violate the protection of free speech.”[14] The ruling left open the question of what the FDA would allow Amarin to say about E-EPA, and in March 2016 the FDA and Amarin agreed that Amarin would submit specific marketing material to the FDA for the FDA to review, and if the parties disagreed on whether the material was truthful, they would seek a judge to mediate.[15]

PAPER

https://link.springer.com/article/10.1023%2FB%3ACONC.0000039128.78645.a8

Synthesis of Fatty-Acid Ethanolamides from Linum catharticum Oils and Cololabis saira Fats
Chemistry of Natural Compounds (Translation of Khimiya Prirodnykh Soedinenii) (2004), 40, (3), 222-226

PAPER

Journal of Molecular Catalysis B: Enzymatic, 84, 173-176; 2012

https://www.sciencedirect.com/science/article/pii/S1381117712000896?via%3Dihub

STARTING MATERIAL CAS 10417-94-4

  • (all-Z)-Δ5,8,11,14,17-Eicosapentaenoic acid
  • (all-cis)-5,8,11,14,17-Eicosapentaenoic acid

PATENT

CN 104846023

https://patents.google.com/patent/CN104846023A/en

Example 1

[0041] A method for preparing a concentrated fish oil fatty acid glycerides, the process steps shown in Figure 1, comprising the steps of:

[0042] S11 using crude enzyme preparation of deep sea fish art: the ratio: (m m) of deep-sea fish through the machine crushed bone formation minced, weighed 600g yue meat, meat by:: water = 0 5.1 water was added seal, in the dark, under nitrogen flow, at 75 ° C cooking lh. Using NaOH to adjust pH to 8.0. Mass fraction of 2% trypsin (trypsin: food grade, Zhengzhou Hong Cheng Chemical Products Limited), in the dark, enzyme 17h at 20 ° C. After 20min by centrifugation 3000r / min, the upper layer was enzymolysis, namely crude fish oil;

[0043] S12 is prepared refined fish oil: Crude fish oil prepared in Step S11 is added a volume ratio of 0.5% phosphoric acid: degummed (crude phosphoric acid fish oil), a concentration of 70% phosphoric acid, followed by centrifugation speed of 3000 rpm / min, and then add a volume ratio of 1% deacidification NaOH, the NaOH concentration is 20%, after centrifugation, the rotational speed of 3000- rpm / min, to obtain refined fish oil;

. [0044] S13 of the refined fish oil fatty acid ethyl ester prepared by esterification process: step S12 is added to the fish oil refining prepared in mass ratio of 0.5% of sodium ethoxide, and a mass ratio of 0.5 in ethanol (ethanol: fish oil refining ), 40 ° C water bath for 1 hour, 1% (by mass) citric acid (citric acid: fish oil refining), standing layer, the upper layer and the liquid was washed with hot deionized water, standing layered repeated three times to give fatty acid ethyl ester.

. [0045] S14 of the fatty acid ethyl ester was extracted Separation: fatty acid ethyl ester obtained in step S13 is subjected to supercritical fluid extraction (extraction process of separation vessel as a rectification column I – separation kettle II), extraction conditions: a rectification column temperature 25-30-35-40 ° C, a pressure of 6 MPa rectification column, separation kettle I temperature 25 ° C, pressure in the separator tank I is 6 MPa, the temperature in the separation tank II 30-45 ° C, C0 2 flow rate of 151,711;

. [0046] S15 of the fatty acid ethyl ester after enzymatic extraction separation processing: The fatty acid ethyl ester obtained in step S14 using Penicillium expansum lipase enzyme, 4% of the amount of enzyme added,, reaction temperature 40 ° C , reaction pH of 10, speed 150 revolutions / min, hydrolysis time 4h, to obtain fatty acid glycerides.

[0047] Example 2

[0048] A process for preparing concentrated fish oil fatty acid glycerides, comprising the steps of:

. [0049] S21 using crude enzyme preparation of deep sea fish art: The procedure of Example 1 with reference to embodiment 11, wherein the cooking temperature is 85 ° C, hydrolysis temperature 25 ° C, centrifuge speed is 4000r / min;

. [0050] S22 refined fish oil preparation: The procedure of Example 1 with reference to embodiment 12; wherein, phosphate: the crude fish oil volume ratio is 1.5%, the phosphoric acid concentration of 75%; K0H: crude fish oil volume ratio of 3%, K0H the concentration of 30%, a centrifugal speed of 4000r / min;

. [0051] S23 of the refined fish oil fatty acid ethyl ester prepared by esterification process: The procedure of Example 1 with reference to embodiment 13; wherein, potassium ethoxide: refined fish oil mass ratio of 1 billion% ethanol: refined fish oil mass ratio of 2.0 , heat the water bath 60 ° C for 3 hours, and acetic acid is acetic acid: refined fish oil mass ratio of 3.0%;

. [0052] S24 was extracted to separate fatty acid ethyl ester: The procedure of Example 1 with reference to embodiment 14; wherein the extraction conditions: temperature rectification column 30-35-40-45 ° C, a pressure rectification column is 15 megabytes Pa, temperature of separation vessel I 35 ° C, pressure in the separator tank I is 8 MPa, the temperature in the separation tank II was 40 ° C, C0 2 flow rate of 171,711;

. [0053] S25 of the fatty acid ethyl ester after enzymatic extraction is carried out the separation treatment: The procedure of Example 1 with reference to embodiment 15; wherein 10% of the amount of enzyme added, reaction temperature 50 ° C, pH 8 hydrolysis, speed 300 rpm / min, hydrolysis time 12h, to obtain fatty acid glycerides.

[0054] Example 3

[0055] – Preparation Method Species of concentrated fish oil fatty acid glycerides, comprising the steps of:

. [0056] S31 using crude enzyme preparation of deep sea fish art: The procedure of Example 1 with reference to embodiment 11, wherein the cooking temperature is 90 ° C, hydrolysis temperature 35 ° C, centrifuge speed is 5000r / min;

. [0057] S32 prepared fine fish oil: The procedure of Example 1 with reference to embodiment 12; wherein, phosphate: the crude fish oil volume ratio of 3% phosphoric acid concentration of 85%; NaOH: crude fish oil volume ratio of 6% and the concentration of NaOH 50%, a centrifugal speed of 5000r / min;

. [0058] S33 of the refined fish oil fatty acid ethyl ester prepared by esterification process: The procedure of Example 1 with reference to embodiment 13; wherein, potassium ethoxide: refined fish oil mass ratio of 1.5%, ethanol: refined fish oil mass ratio of 4.0 heat treatment is 80 ° C water bath for 5 hours, citric acid and citric acid are added: refined fish oil mass ratio of 5.0%;

. [0059] S34 was extracted to separate fatty acid ethyl ester: The procedure of Example 1 with reference to embodiment 14; wherein the extraction conditions: temperature rectification column 30-35-40-45 ° C, pressure column 17 trillion Pa, I of separation vessel temperature 40 ° C, pressure in the separator tank I is 10 MPa, the temperature in the separation tank II is 45 ° C, C0 2 flow rate is? L / h;

. [0060] S35 of the fatty acid ethyl ester after enzymatic extraction separation processing: The procedure of Example 1 with reference to embodiment 15; wherein 20% of the amount of enzyme added, reaction temperature 60 ° C, a pH of 6.5 hydrolysis, speed 300 rpm / min, hydrolysis time 24h, to obtain fatty acid glycerides.

[0061] Comparative Example

[0062] S1 • obtaining crude fish: The procedure of Example 1 with reference to embodiment 11;

. [0063] S2 refined fish oil preparation: see Example 1, Step 12;

. [0064] S3 of refined fish oil fatty acid ethyl ester prepared by esterification process: Step 1, Example 13 process embodiment with reference, to obtain fatty acid ethyl ester.

PATENT

https://patents.google.com/patent/WO2014054435A1

WO 2014054435

 In recent years, highly unsaturated fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been clarified for their pharmacological effects and are used as raw materials for pharmaceuticals and health foods. Since these polyunsaturated fatty acids have a plurality of double bonds, it is not easy to obtain them by chemical synthesis. Therefore, most of industrially used highly unsaturated fatty acids are produced by extraction or purification from marine organism-derived materials rich in polyunsaturated fatty acids, such as fish oil, etc. However, the content of highly unsaturated fatty acid is not necessarily high, because the biological material is a mixture of various kinds of fatty acids having different numbers of carbon atoms, number and position of double bonds, constitutional ratio of stereoisomers, and the like. For this reason, conventionally, it has been required to selectively purify a target highly unsaturated fatty acid from a biological raw material.
 Patent Document 1 discloses a supercritical gas extraction method after a thin film distillation method when a raw material containing a highly unsaturated fatty acid or an alkyl ester thereof is treated by a thin film distillation method, a supercritical gas extraction method and a urea addition method A method for purifying a highly unsaturated fatty acid or an alkyl ester thereof is described.
 In Patent Document 2, a raw material containing a highly unsaturated fatty acid such as EPA is subjected to vacuum precision distillation treatment, and the resulting EPA or a fraction containing a lower alcohol ester thereof is mixed with an aqueous silver nitrate solution, whereby a high purity eicosapentaene A method of purifying an acid or a lower alcohol ester thereof is described. It is described that the condition of the vacuum precision distillation is a pressure of 5 mmHg (665 Pa) or less, preferably 1 mmHg (133 Pa) or less, 215 ° C. or less, preferably 210 ° C. or less.
 Further, Patent Document 3 discloses a process for producing eicosapentaenoic acid or an ester thereof having a concentration of 80% or more by gradually distilling a raw material containing a highly unsaturated fatty acid or an alkyl ester thereof using a distillation tower having three or more stages Is described. It is described that the condition of the distillation is 10 Torr (1330 Pa) or less, preferably 0.1 Torr (13.3 Pa) or less, 210 ° C. or less, preferably 195 ° C. or less.
 However, highly unsaturated fatty acids having higher concentrations and purities than those obtained by the above-mentioned conventional methods are required as raw materials for pharmaceuticals and health foods.
There are cis and trans isomers in highly unsaturated fatty acids. Most of the highly unsaturated fatty acids in vivo are cis, however, they may be converted from cis form to trans form by heating or the like at the stage of purification from biological origin materials (Non-Patent Document 1). Therefore, polyunsaturated fatty acids conventionally purified industrially from biologically derived raw materials contain a certain amount of trans isomer. However, trans fatty acids have been reported to increase health risks, especially LDL cholesterol levels, and increase the risk of cardiovascular disease. In the United States and Canada, foods are obliged to indicate the content of trans fatty acids.
 Therefore, there is a need for a highly unsaturated fatty acid-containing composition which not only contains the targeted highly unsaturated fatty acid at a high concentration as a raw material for pharmaceuticals and health foods but also contains a trans fatty acid content as low as possible . However, conventionally, purification of highly unsaturated fatty acids has not been conducted focusing on the stereoisomer ratio.
Patent Document 1: Japanese Patent Application Laid-Open No. 10-95744
Patent Document 2: Japanese Patent Application Laid-Open No. 7-242895
Patent Document 3: Japanese Patent No. 3005638

Non-patent literature

[0010]
Non-patent document 1: Journal of the American Oil Chemists’ Society, 1989, 66 (12): 1822-1830

Example 

[0035]
 Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited to only these examples.

[0036]
 In the following examples, the method of composition analysis of highly unsaturated fatty acids and the method of quantitating stereoisomers are as follows.
9 μL of the measurement sample was diluted to 1.5 mL of n-hexane, and the content ratio of each fatty acid and the content ratio of isomers were analyzed using a gas chromatography analyzer (Type 6890 GC, manufactured by Agilent Technologies) under the following conditions did. The results are expressed as mass% converted from the area of the chromatogram.
<Column condition>
Column: DB-WAX 0.25 mm × 30 m manufactured by J & W Co., column temperature: 210 ° C.
He flow rate: 1.0 ml / min, He pressure: 134 kPa
<Detection condition>
2 flow rate: 30 ml / min, Air flow rate : 400 ml / min
He flow rate: 10 ml / min, DET temperature: 260 ° C.
The isomer ratio in the target highly unsaturated fatty acid was obtained by the following formula.

[0037]
[Expression 1]

[0038]
(Example 1)
Raw material: 1000 mL of anhydrous ethanol solution in which 50 g of sodium hydroxide was dissolved was added to 1 kg of sardine oil, mixed and stirred at 70 to 80 ° C. for 1 hour, then 500 mL of water was added and mixed well, 1 It was left standing for a while. The separated aqueous phase was removed and the oil phase was washed several times with water to neutralize the washings to give 820 g of ethyl esterified sardine oil.
As shown in Table 1, the composition of the sardine oil was 44.09% (mass%, hereinafter the same) of eicosapentaenoic acid (EPA), 1.52% of eicosatetraenoic acid (ETA), 1.52% of arachidonic acid (AA) 1.77%, docosahexaenoic acid (DHA) 6.92%. Also, the trans isomer ratio in EPA was 1.23%.
Step (1) 160 ml of n-hexane was added to 300 g of the ethyl esterified sardine oil prepared above, and the mixture was stirred well and dissolved. To this was added 500 mL of an aqueous solution containing 50% by weight of silver nitrate, and the mixture was stirred under conditions of 5 to 30 ° C. After standing, the separated n-hexane phase was removed, and the aqueous phase was recovered.
Step (2): 2000 mL of fresh n-hexane was added to the aqueous phase obtained in the step (1), and the mixture was sufficiently stirred at 50 to 69 ° C. to extract the fatty acid ethyl ester into n-hexane. After standing, the separated aqueous phase was removed and the n-hexane phase was concentrated. The crude fatty acid ethyl ester crude product contained in this n-hexane phase contained 74.54% EPA, 0.32% ETA, 0.17% AA and 14.87% DHA in total fatty acids as shown in Table 1 It was. Also, the trans isomer ratio in EPA was 0.19%.
Step (3): The n-hexane phase containing the fatty acid ethyl ester obtained in the step (2) was maintained under conditions of a top vacuum degree of 1 Pa or less and a distillation temperature of 170 to 190 ° C. using a packed tower precision distillation apparatus While performing vacuum distillation to obtain a highly purified EPA ethyl ester-containing composition in a yield of about 60%. As shown in Table 1, this EPA ethyl ester-containing composition contained 98.25% of EPA, 0.43% of ETA, 0.21% of AA, and 0.05% of DHA in total fatty acids. Also, the trans isomer ratio in EPA was 0.45%.
The yield of EPA in this example in which the steps were performed in the order of (1), (2), (3) was about 53%.

[0039]
Example 2 The
steps (1), (2) and (3) were carried out in the same manner as in Example 1 except that the step (3) was carried out while maintaining the distillation temperature of 180 to 185 ° C., EPA ethyl ester-containing composition was obtained in a yield of about 58%. As shown in Table 1, this EPA ethyl ester-containing composition contained 98.29% of EPA, 0.40% of ETA, 0.32% of AA, and 0.05% of DHA in total fatty acids. Also, the trans isomer ratio in EPA was 0.28%, and the trans isomer was extremely small.
Comparative Example 1 An
EPA ethyl ester-containing composition was obtained in the same manner as in Example 1, except that the top vacuum degree was set to 13.3 Pa (0.1 Torr) in the step (3). As shown in Table 1, the composition contained EPA content ratio as high as 97.44% in the total fatty acid, but the trans isomer ratio in EPA was high (1.37%).

[0040]
Comparative Example 2 The
EPA ethyl ester-containing composition was obtained by performing vacuum distillation (step (3)) of ethyl esterified sardine oil and then steps (1) and (2). The conditions of each step were the same as in Example 1. As shown in Table 1, this composition contained 95.05% EPA, 0.72% ETA, 0.50% AA, 0.21% DHA in total fatty acids, the trans isomer ratio in EPA was 1.55% Met. The yield of EPA in this comparative example in which the steps were carried out in the order of (3), (1) and (2) was about 31%, and the EPA yield greatly decreased as compared with Example 1.
By changing the condition of the vacuum distillation in this Comparative Example (0.5 Pa, 185 to 195 ° C.), it was possible to raise the content of EPA in the total fatty acids in the composition to 98.12%, however, The rate further declined and the trans isomer ratio in EPA was 2.01%, further increased.

[0041]
[table 1]

[0042]
Examples 3 to 4 and Comparative Example 3 In the
step (3), the distillation temperature was 180 ° C. (Example 3), 190 ° C. (Example 4), 200 ° C. (Comparative Example 3), and the vacuum distillation time was A highly purified EPA ethyl ester-containing composition was obtained in the same manner as in Example 1 except that various changes were made and the trans isomer ratio of EPA in the composition was determined. The results are shown in Fig. 1. 1, in Examples 3 to 4 having a distillation temperature of 190 ° C. or less, the trans isomer ratio was less than 1% by mass, but in Comparative Example 3 having a distillation temperature of 200 ° C., the trans isomer The ratio exceeds 1% by mass.

References

  1. Jump up to:a b c d e f g h Icosapent ethyl Label Last revised June 2015. Check for updates at FDA label index page here
  2. Jump up to:a b c Jacobson TA, et al, NLA Expert Panel. National Lipid Association Recommendations for Patient-Centered Management of Dyslipidemia: Part 2. J Clin Lipidol. 2015 Nov-Dec;9(6 Suppl):S1-S122.e1. PMID 26699442 Free full text
  3. Jump up to:a b Weintraub, HS (2014). “Overview of prescription omega-3 fatty acid products for hypertriglyceridemia”Postgrad Med126: 7–18. doi:10.3810/pgm.2014.11.2828PMID 25387209. Retrieved 20 April 2015.
  4. Jump up^ University of Utah Pharmacy Services (15 August 2007) “Omega-3-acid Ethyl Esters Brand Name Changed from Omacor to Lovaza”
  5. Jump up^ Omtryg Label Revised April 2014
  6. Jump up^ FDA Omega-3 acid ethyl esters products Page accessed 31 March 2016
  7. Jump up^ “Epanova (omega-3-carboxylic acids)”CenterWatch. Retrieved 15 December 2014.
  8. Jump up to:a b Ito MK. A Comparative Overview of Prescription Omega-3 Fatty Acid Products. P T. 2015 Dec;40(12):826-57. PMID 26681905 Free PMC Article PMC 4671468
  9. Jump up^ Sweeney MET. Hypertriglyceridemia Pharmacologic Therapy for Medscape Drugs & Diseases, Ed. Khardori R. Updated: 14 April 2015, page accessed 1 April 2016
  10. Jump up^ CenterWatch Vascepa (icosapent ethyl) Page accessed 31 March 2016
  11. Jump up^ VHA Pharmacy Benefits Management Strategic Healthcare Group and the Medical Advisory Panel. October 2005 National PBM Drug Monograph Omega-3-acid ethyl esters (Lovaza, formerly Omacor)
  12. Jump up^ Matthew Herper for Forbes. 17 October 2013 Why The FDA Is Right To Block Amarin’s Push To Market Fish Oil To Millions
  13. Jump up^ Thomas, Katie (7 May 2015). “Drugmaker Sues F.D.A. Over Right to Discuss Off-Label Uses”New York Times. Retrieved 17 May 2017.
  14. Jump up^ Andrew Pollack for the New York Times. 7 August 2015 Court Forbids F.D.A. From Blocking Truthful Promotion of Drug
  15. Jump up^ Katie Thomas for the New York Times. 8 March 2016 F.D.A. Deal Allows Amarin to Promote Drug for Off-Label Use
CN1288732A *2000-07-122001-03-28刘玉Soft concentrated fish oil capsule and its supercritical CO2 extraction and rectification process
CN101255380A *2007-03-032008-09-03苑洪德Triglyceride type fish oil and method for making same
CN101818176A *2010-04-092010-09-01浙江兴业集团有限公司;华南理工大学Method for transforming fatty acid ethyl ester into glyceride
CN102964249A *2012-11-162013-03-13成都圆大生物科技有限公司Process capable of simultaneously producing and separating high-purity EPA (eicosapentaenoic acid) ethyl ester and high-purity DHA (docosahexaenoic acid) ethyl ester
CN102994236A *2012-12-112013-03-27成都圆大生物科技有限公司Method for preparing fatty acid ethyl ester with Omega-3 content of more than 90 percent
Ethyl eicosapentaenoic acid
Ethyl eicosapentaenoate.png
Names
IUPAC name

Ethyl (5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoate
Other names

Eicosapentaenoic acid ethyl ester; Ethyl eicosapentaenoate; Eicosapent; Icosapent ethyl; EPA ethyl ester; E-EPA
Identifiers
3D model (JSmol)
ChEBI
ChemSpider
PubChem CID
Properties
C22H34O2
Molar mass 330.51 g·mol−1
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

////////////Icosapent ethyl, fda 2012, Timnodonic acid ethyl ester, Vascepa, AMR 101, AMR-101, E-EPA, Ethyl eicosapentaenoic acid , Fast-track status, Orphan drug designation 

CCOC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CC

Revefenacin, ревефенацин , ريفيفيناسين , 瑞维那新 ,


Revefenacin.png

Revefenacin; 864750-70-9; TD-4208; UNII-G2AE2VE07O; G2AE2VE07O; TD-4208; GSK-1160724;

160724; GSK 1160724; TD-4028; YUPELRI

Molecular Formula: C35H43N5O4
Molecular Weight: 597.76 g/mol

[1-[2-[[4-[(4-carbamoylpiperidin-1-yl)methyl]benzoyl]-methylamino]ethyl]piperidin-4-yl] N-(2-phenylphenyl)carbamate

TD-4208
UNII:G2AE2VE07O
ревефенацин [Russian] [INN]
ريفيفيناسين [Arabic] [INN]
瑞维那新 [Chinese] [INN]

Revefenacin is under investigation for the treatment of Chronic Obstructive Pulmonary Disease (COPD).

  • Originator Theravance
  • Developer Theravance Biopharma
  • Class Antiasthmatics; Biphenyl compounds; Carbamates; Piperidines
  • Mechanism of Action Muscarinic receptor antagonists
  • Preregistration Chronic obstructive pulmonary disease
  • 17 Sep 2018 Efficacy data from two replicate 12-week phase III trials and a 12-month safety trial in Chronic obstructive pulmonary disease (COPD) presented at the European Respiratory Society International Congress (ERS-2018)
  • 31 May 2018 Theravance Biopharma in collaboration with Theravance Biopharma initiates enrolment in a phase III trial for Chronic obstructive pulmonary disease in USA (NCT03573817)
  • 18 May 2018Efficacy and adverse events data from a phase I trial in Chronic obstructive pulmonary disease presented at the 114th International Conference of the American Thoracic Society

The compound was licensed to GlaxoSmithKline by Theravance for the inhalation treatment of chronic obstructive pulmonary disease in 2004. The rights were returned in 2009. In 2014, Theravance Biopharma spun-off from Theravance. In 2015, Theravance Biopharma and Mylan enter in a co development agreement for the global development and commercialization of the once-daily nebulizer for the treatment of chronic obstructive pulmonary disease and other respiratory diseases.

SYN

WO 2012009166

SYN OF INT

STR1

FINAL

STR1

PAPER
Discovery of (R)-1-(3-((2-Chloro-4-(((2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl)amino)methyl)-5-methoxyphenyl)amino)-3-oxopropyl)piperidin-4-yl (1,1′-biphenyl)-2-ylcarbamate (TD-5959, GSK961081, batefenterol): First-in-class dual pharmacology multivalent muscarinic antagonist and 2 agonist (MABA) for the treatment of chronic obstructive pulmonary disease (COPD)
J Med Chem 2015, 58(6): 2609

Discovery of (R)-1-(3-((2-Chloro-4-(((2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl)amino)methyl)-5-methoxyphenyl)amino)-3-oxopropyl)piperidin-4-yl [1,1′-Biphenyl]-2-ylcarbamate (TD-5959, GSK961081, Batefenterol): First-in-Class Dual Pharmacology Multivalent Muscarinic Antagonist and β2 Agonist (MABA) for the Treatment of Chronic Obstructive Pulmonary Disease (COPD)

Departments of Medicinal Chemistry, Pharmacology, §Drug Metabolism and Pharmacokinetics, and Molecular and Cellular Biology, Theravance Biopharma, Inc., 901 Gateway Boulevard, South San Francisco, California 94080, United States
J. Med. Chem.201558 (6), pp 2609–2622
DOI: 10.1021/jm501915g
*Phone: 650-808-3737. E-mail: ahughes@theravance.com
Abstract Image

Through application of our multivalent approach to drug discovery we previously reported the first discovery of dual pharmacology MABA bronchodilators, exemplified by 1. Herein we describe the subsequent lead optimization of both muscarinic antagonist and β2 agonist activities, through modification of the linker motif, to achieve 24 h duration of action in a guinea pig bronchoprotection model. Concomitantly we targeted high lung selectivities, low systemic exposures and identified crystalline forms suitable for inhalation devices. This article culminates with the discovery of our first clinical candidate 12f (TD-5959, GSK961081, batefenterol). In a phase 2b trial, batefenterol produced statistical and clinically significant differences compared to placebo and numerically greater improvements in the primary end point of trough FEV1 compared to salmeterol after 4 weeks of dosing in patients with moderate to severe chronic obstructive pulmonary disease (COPD).

PATENT

WO 2006099165

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2006099165

FIG. 18 shows a PXRD pattern of Form I of the crystalline freebase of the compound of formula I. This crystalline freebase is further characterized by the DSC trace in FIG. 19, the TGA trace in FIG. 20, the DMS trace in FIG. 21, and the micrographic image in FIG. 22.
FIG. 23 shows a PXRD pattern of Form II of the crystalline freebase of the compound of formula I. This crystalline freebase is further characterized by the DSC trace in FIG. 24, the TGA trace in FIG. 25, and the DMS trace in FIG. 26.

PREPARATION 1
Biphenyl-2-ylcarbamic Acid Piperidin-4-yl Ester
Biphenyl-2-isocyanate (97.5 g, 521 mmol) and 4-hydroxy-N-benzylpiperidine (105 g, 549 mmol) were heated together at 70 0C for 12 hours. The reaction mixture was then cooled to 50 0C and ethanol (1 L) was added and then 6M HCl (191 mL) was added slowly. The resulting mixture was then cooled to ambient temperature and ammonium formate (98.5 g, 1.56 mol) was added and then nitrogen gas was bubbled through the solution vigorously for 20 minutes. Palladium on activated carbon (20 g, 10 wt% dry basis) was then added and the reaction mixture was heated at 40 0C for 12 hours, and then filtered through a pad of Celite. The solvent was then removed under reduced pressure and IM HCl (40 mL) was added to the crude residue. The pH of the mixture was then adjusted with IO N NaOH to pH 12. The aqueous layer was extracted with ethyl acetate (2 x 150 mL) and the organic layer was dried (magnesium sulfate), filtered and the solvent removed under reduced pressure to give 155 g of the title intermediate (100% yield). HPLC (10-70) Rt = 2.52; m/z: [M + H+] calc’d for C18H20N2O2 297.15; found 297.31
PREPARATION 2
iV-Benzyl-iV-methylaminoacetaldehvde
To a 3-necked 2-L flask was added N-benzyl-N-methylethanolamine (30.5 g, 0.182 mol), DCM (0.5 L), DIPEA (95 mL, 0.546 mol) and DMSO (41 mL, 0.728 mol).

Using an ice bath, the mixture was cooled to about -10 °C and sulfur trioxide pyridine-complex (87 g, 0.546 mol) was added in 4 portions over 5 minute intervals. The reaction was stirred at -10 0C for 2 hours. Before removing the ice-bath, the reaction was quenched by adding water (0.5 L). The aqueous layer was separated and the organic layer was washed with water (0.5 L) and brine (0.5 L) and then dried over magnesium sulfate and filtered to provide the title compound which was used without further purification.
PREPARATION 3
Biphenyl-2-ylcarbamic Acid l-[2-(Εenzylmethylammo)ethyllpiperidin-4-yl Ester
To a 2-L flask, containing the product of Preparation 2 in DCM (0.5 L) was added the product of Preparation 1 (30 g, 0.101 mol) followed by sodium triacetoxyborohydride (45 g, 0.202 mol). The reaction mixture was stirred overnight and then quenched by the addition of 1 N hydrochloric acid (0.5 L) with vigorous stirring. Three layers were observed and the aqueous layer was removed. After washing with IN NaOH (0.5 L)3 a homogenous organic layer was obtained which was then washed with a saturated solution of aqueous NaCl (0.5 L), dried over magnesium sulfate, filtered and the solvent removed under reduced pressure. The residue was purified by dissolving it in a minimal amount of isopropanol and cooling this solution to 0 °C to form a solid which was collected and washed with cool isopropanol to provide 42.6 g of the title compound (95% yield). MS m/z: [M + H+] calc’d f for C28H33N3O2444.3; found 444.6. Rf=3.5l min (10-70 ACN:H2O, reverse phase HPLC).
PREPARATION 3 A
Biphenyl-2-ylcarbamic Acid l-f2-(Benzylmethylammo)ethyllpiperidin-4-yl Ester
The title compound was prepared by mesylation of iV-benzyl-N-methyl
ethanolamine, which was then reacted with biphenyl-2-ylcarbamic acid piperidin-4-yl ester in an alkylation reaction.
A 500 mL flask (reactor flask) was charged with N-benzyl-iV-methylethanolamine (24.5 mL), DCM (120 mL), NaOH (80 mL; 30wt%) and tetrabutylammonium chloride. Mixing at low speed throughout the reaction, the mixture was cooled to -10 °C (cooling bath), and the addition funnel charged with DCM (30 mL) and mesyl chloride (15.85 mL), which was added drop wise at a constant rate over 30 minutes. The addition was exothermic, and stirring was continued for 15 minutes while the temperature equilibrated back to -10 0C. The reaction was held for at least 10 minutes to ensure full hydrolysis of the excess mesyl chloride.
A 250 mL flask was charged with biphenyl-2-ylcarbamic acid piperidin-4-yl ester (26 g; prepared as described in Preparation 1) and DCM (125 mL), stirred for 15 minutes at room temperature, and the mixture chilled briefly to 10 0C to form a slurry. The slurry was then charged into the reactor flask via the addition funnel. The cooling bath was removed and the reaction mixture was warmed to 5 °C. The mixture was transferred to a separatory funnel, the layers allowed to settle, and the aqueous layer removed. The organic layer was transferred back to the reactor flask, stirring resumed, the mixture held to room
temperature, and the reaction monitored by HPLC for a total of 3.5 hours.
The reactor flask was charged with NaOH (IM solution; 100 mL), stirred, and the layers allowed to settle. The organic layer was separated, washed (NaCl satd. solution), its volume partially reduced under vacuum, and subjected to repeated IPA washings. The solids were collected and allowed to air-dry (25.85 g, 98% purity). Additional solids were obtained from further processing of the mother liquor (volume reduction, EPA, cooling).
PREPARATION 4
Biphenyl-2-ylcarbamic Acid l-(2-Methylaminoethyl)piperidin-4-yl Ester
To a Parr hydrogenation flask was added the product of Preparation 3 (40 g, 0.09 mol) and ethanol (0.5 L). The flask was flushed with nitrogen gas and palladium on activated carbon (15g, 10 wt% (dry basis), 37% wt/wt) was added along with acetic acid (20 mL). The mixture was kept on the Parr hydrogenator under a hydrogen atmosphere (-50 psi) for 3 hours. The mixture was then filtered and washed with ethanol. The filtrate was condensed and the residue was dissolved in a minimal amount of DCM. Isopropyl acetate (10 volumes) was added slowly to form a solid which was collected to provide 22.0 g of the title compound (70% yield). MS m/z: [M + H+] calc’d for C21H27N3O2 354.2; found 354.3. R/=2.96 min (10-70 ACNrH2O, reverse phase HPLC).
PREPARATION 5
Biphenyl-2-ylcarbamic Acid l-{2-[(4-Formylbenzoyr)
methylaminol ethyll piperidin-4- yl Ester
To a three-necked 1-L flask was added 4-carboxybenzaldehyde (4.77 g,
31.8 mmol), EDC (6.64 g, 34.7 mmol), HOBT (1.91 g, 31.8 mmol), and DCM (200 mL). When the mixture was homogenous, a solution of the product of Preparation 4 (10 g, 31.8 mmol) in DCM (100 mL) was added slowly. The reaction mixture was stirred at room temperature for approximately 16 hours and then washed with water (1 x 100 mL), IN HCl (5 x 60 mL), IN NaOH (1 x 100 mL) brine (1 x 5OmL)3 dried over sodium sulfate, filtered and concentrated to afford 12.6 g of the title compound (92% yield; 85% purity based on HPLC). MS m/z: [M + H+] calc’d for C29H31N3O4 486.2; found 486.4. i?y=3.12 min (10-70 ACNiH2O, reverse phase HPLC).
EXAMPLE 1
Biphenyl-2-ylcarbamic Acid 1 -(2- { |4-(4-Carbamoylpiperidin- 1 -ylmethvD
benzoylimethylamino) ethyl’)piperidin-4-vl Ester

To a three-necked 2-L flask was added isonipecotamide (5.99 g, 40.0 mmol), acetic acid (2.57 mL), sodium sulfate (6.44 g) and isopropanol (400 mL). The reaction mixture was cooled to 0-10 0C with an ice bath and a solution of biphenyl-2-ylcarbamic acid l-{2-[(4-formylbenzoyl)methylamino]ethyl}piperidin-4-yl ester (11 g, 22.7 mmol; prepared as described in Preparation 5) in isopropanol (300 mL) was slowly added. The reaction mixture was stirred at room temperature for 2 hours and then cooled to 0-10 0C. Sodium triacetoxyborohydride (15.16 g, 68.5 mmol) was added portion wise and this mixture was stirred at room temperature for 16 hours. The reaction mixture was then concentrated under reduced pressure to a volume of about 50 mL and this mixture was acidified with IN HCl (200 mL) to pH 3. The resulting mixture was stirred at room temperature for 1 hour and then extracted with DCM (3 x 250 mL). The aqueous phase was then cooled to 0-5 °C with an ice bath and 50% aqueous NaOH solution was added to adjust the pH of the mixture to 10. This mixture was then extracted with isopropyl acetate (3 x 300 mL) and the combined organic layers were washed with water (100 mL), brine (2 x 50 mL), dried over sodium sulfate, filtered and concentrated to afford 10.8 g of the title compound (80% yield. MS m/z: [M + H+] calc’d for C35H43N5O4 598.3; found 598.6. Rj=232 min (10-70 ACNiH2O, reverse phase HPLC).

EXAMPLE 2
Crystalline Diphosphate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4- Carbamoylpiperidin-l-ylmethyl)benzoyl1methylamino>ethyDpiperidin-4-yl Ester
500 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiρeridin-l-ylmethyl) benzoyl]methylamino}ethyl)piperidin-4-yl ester (0.826 mmol of 96% pure material;
prepared as described in Example 1) was taken up in 5 ml of water and 1.5 ml of IM phosphoric acid. The pH was adjusted to approximately pH 5.3 with an additional 0.25ml of IM phosphoric acid (equaling 2.1 molar equivalents). The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness to yield an amorphous diphosphate salt.
20 mg of the amorphous diphosphate salt was dissolved in 2 ml of IPA: ACN (1:1). 0.1 ml of water was added and the mixture heated to 60 °C under stirring. Almost all of the solids dissolved. The suspension was allowed to cool to ambient temperature, under stirring, overnight. The resulting crystals were collected by filtration and air-dried for 20 minutes to give the title compound (18.5 mg, 93% yield) as a white crystalline solid.
When examined under a microscope using polarized light, the crystals exhibited some birefringence.
EXAMPLE 3
Crystalline Diphosphate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{|4-(4- Carbamoylpiperidin-l-vhτiethyl)benzoyl]methylamino}ethyl)piperidin-4-yl Ester
5.0 g of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (freebase; prepared as described in Example 1) was combined with 80 ml of IPA:ACN (1:1). 4.0 ml of water was added and the mixture heated to 50 °C under stirring, forming a clear solution. To this was added dropwise at 50 °C, 16 ml IM phosphoric acid. The resulting cloudy solution was stirred at 50 °C for 5 hours, then allowed to cool to ambient temperature, under slow stirring, overnight. The resulting crystals were collected by filtration and air-dried for 1 hour, then under vacuum for 18 hours, to give the title compound (5.8 g, 75% yield) as a white crystalline solid (98.3% purity by HPLC).

EXAMPLE 4
Crystalline Monosulfate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4- Carbamoylpiperidm-l-ylmethvπbenzoyllmethylamino>ethyl)piperidm-4-yl Ester
442 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-Carbamoylpiperidin-l-ylmethyl) benzoyl]methylamino} ethyl)piperidin-4-yl ester (0.739 mmol of 96% pure material;
prepared as described in Example 1) was taken up in 5 ml of H2OrACN (1 : 1) and 1.45 ml of IN sulfuric acid was added slowly, while monitoring the pH. The pH was adjusted to approx. pH 3.3. The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness to yield a monosulfate salt.
30.3 mg of the monosulfate salt was dissolved in 1.65 ml of IPA:ACN (10:1). The suspension was heated by placing the vial in a pre-heated 60 °C water bath for 30 minutes. A viscous material was formed and the heat increased to 70 °C for 30 minutes. Since the material remained viscous, the heat was lowered to 60 0C and the mixture heated for an additional hour. The heat was turned off and the mixture was allowed to cool to room temperature. After 4 days, the material appeared to be solid, and the sample was allowed to sit for an additional nine days. The solid was then filtered and dried using a vacuum pump for 1 hour to give the title compound (23 mg, 76% yield).
EXAMPLE 5
Crystalline Monosulfate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{[~4-(4- Carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino>ethyl)piperidin-4-yl Ester
161 g of the monosulfate salt (prepared as described in Example 4) was dissolved in 8.77 ml of IPA:ACN (10:1). The suspension was heated by placing the vial in a pre-heated 70 °C water bath for 1.5 hours. Oil droplets formed within 5 minutes. The heat was lowered to 60 °C and the mixture heated for an additional 1.5 hours, followed by heating at 50 °C for 40 minutes, at 40 °C for 40 minutes, then at 30 0C for 45 minutes. The heat was turned off and the mixture was allowed to slowly cool to room temperature. The next day, the material was viewed under a microscope and indicated needles and plates. The material was then heated at 40 °C for 2 hours, at 35 0C for 30 minutes, and then at 30 °C for 30 minutes. The heat was turned off and the mixture was allowed to slowly cool to room temperature. The solid was then filtered and dried using a vacuum pump for 1 hour to give the title compound (117 mg, 73% yield).

EXAMPLE 6
Crystalline Dioxalate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{|4-(4-Carbamoylpiperidin- 1 -ylmethyl)benzoyl]methylamino> ethyl)piperidin-4-yl Ester
510 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino} ethyl)piperidin-4-yl ester (0.853 mmol of 96% pure material; prepared as described in Example 1) was taken up in 5 ml of H2O:ACN (1:1) and 1.7 ml of IM aqueous oxalic acid was added slowly, while monitoring the pH. The pH was adjusted to approx. pH 3.0. The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness to yield a dioxalate salt.
31.5 mg of the dioxalate salt was dissolved in 2.76 ml of 94%IPA/6%H20. The mixture was stirred in a pre-heated 60 °C water bath for 2.5 hours. After 25 minutes, all of the sample was in solution. The heat was turned off and the mixture was allowed to cool to room temperature. The next day, a small amount of viscous material was present. The vial was refrigerated at 4 °C. After 4 days, the viscous material was still present. The vial was then placed at room temperature and observed one month later. The material appeared to be solid, and was observed to be crystalline under a microscope. The solid was then filtered and dried using a vacuum pump for 1 hour to give the title compound (20 mg, 63.5% yield).
EXAMPLE 7
Crystalline Dioxalate Salt of Biphenyl-2-ylcarbamic Acid l-(2-{T4-(4-Carbamoylpiperidin- 1 -ylmethyl)benzoyl]methylammo) ethvDpiperidin-4-yl Ester
150 mg of the dioxalate salt (prepared as described in Example 6) was dissolved in 13.1 ml of 94%IPA/6%H20. The mixture was stirred in a pre-heated 60 °C water bath for 2.5 hours. The heat was turned off and the mixture was allowed to cool to room
temperature. The vial was refrigerated at 4 °C. After 6 days, an oily material was observed with what appeared to be a crystal on the side of the vial. The vial was then allowed to reach room temperature, at which point seeds (crystalline material from Example 6) were added and allowed to sit for 16 days. During this time, more crystals were observed to come out of solution. The solid was then filtered and dried using a vacuum pump for 14 hours to give the title compound (105 mg, 70% yield).

EXAMPLE 8
Crystalline Freebase Biphenyl-2-ylcarbamic Acid l-(2-(f4-(4-Carbamoylpiperidin-l- ylmethvDbenzoyl]methylaniino}ethyl)piperidin-4-yl Ester (Form T)
109 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (prepared as described in
Example 1) was dissolved in 0.56 ml of H2O: ACN (1:1). The suspension was left in a vial (cap loosely placed on top) to allow for a slower evaporation time. The vial was placed under a nitrogen flow environment, although the nitrogen was not used for evaporation, only for the environment. A precipitate was visible within 1 day, which was observed to be crystalline under a microscope. The solid was then placed on a high vacuum line to remove all solvent to give the title compound. Quantitative recovery, 97.8% pure by HPLC.

In an alternate procedure, after dissolving in H2O: ACN (1:1) (approximately 350 mg/mL), the vial was stored at 5 0C, and the precipitate was visible at day 2. The solid was filtered, rinsed with water, and dried on high vacuum overnight. Recovery was 55%, with the solid having 98.2% purity and the liquid having 92.8% purity.
EXAMPLE 9
Crystalline Freebase Biphenyl-2-ylcarbamic Acidl-(2-{J4-(4-Carbamoylpiperidin- l-yhiaethyl)benzoyllmethylammo|ethvDpiperidin-4-yl Ester (Form T)
50.4 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (prepared as described in
Example 1) was dissolved in 0.144 ml of H2O:ACN (1:1). The suspension was left in vial (cap loosely placed on top) to allow for a slower evaporation time. The vial was refrigerated at 4 0C for 6 days. A precipitate was visible after 2 days. The solid was filtered and placed on a high vacuum line to remove all solvent and give the title compound as a white solid (27.8 mg, 55.2 % yield).
EXAMPLE 10
Crystalline Freebase Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4-Carbamoylpiperidin- l-vhnethvDbenzoyl]methylamino>ethvDpiperidin-4-yl Ester (Form T)
230 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-yhnethyl)benzoyl]methylamino}ethyl)piρeridin-4-yl ester (prepared as described in
Example 1) was dissolved in 0.2 ml of H2O:ACN (1:1), using slight heat. The mixture was then heated in a 70 °C water bath for 2 hours. The heat was turned off and the mixture was allowed to cool to room temperature, then refrigerated at 4 °C for 1 hour. 50 μl of water was then added (oiled out), followed by the addition of 40 μl of ACN to get the sample back into solution. Seeds (crystalline material from Example 8) were added under slow stirring at room temperature. Crystals started to form ,and the mixture was allowed to sit overnight, with slow stirring. The next day, a heat cool cycle was applied (30 °C for 10 minutes, 40 0C for 10 minutes, then 50 °C for 20 minutes). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, a second heat/cool cycle was applied (60 0C for 1 hour, with dissolving observed at 70 °C). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, crystals were present and a third heat cool cycle was applied (60 0C for 3 hours). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, a heat cool cycle was applied (60 °C for 3 hours, slow cool, then 60 °C for 3 hours). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. After 3 days, the solid was filtered and placed on a high vacuum line to remove all solvent and give the title compound.
EXAMPLE 11
Crystalline Freebase Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4-Carbamoylpiperidin- l-ylmethyl)benzoyl]methylamino|ethyl)piperidin-4-yl Ester (Form JD
70 mg of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-yhnethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (prepared as described in
Example 1) was dissolved in 0.1 mL ACN. After addition of 0.3 ml MTBE, the solution appeared cloudy. An additional 50 μl of ACN was added to clarify the solution (155 mg/ml ACN:MTBE = 1 :2). The mixture was left in the vial and capped. Crystals appeared by the next day. The solid was then filtered and placed on a high vacuum line to remove all solvent and give the title compound.

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2011008809

U.S. Patent Publication No. 2005/0203133 to Mammen et al. discloses novel biphenyl compounds that are expected to be useful for treating pulmonary disorders such as chronic obstructive pulmonary disease (COPD) and asthma. In particular, the compound biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl) benzoyl]methylamino}ethyl)piperidin-4-yl ester is specifically described in this application as possessing muscarinic receptor antagonist or anticholinergic activity.

The chemical structure of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoyl piperidin- 1 -ylmethyl)benzoyl]methylamino } ethyl)piperidin-4-yl ester is represented by formula I:

I

The compound of formula I has been named using the commercially-available AutoNom software (MDL, San Leandro, California).

Therapeutic agents useful for treating pulmonary or respiratory disorders are advantageously administered directly into the respiratory tract by inhalation. In this regard, several types of pharmaceutical inhalation devices have been developed for administering therapeutic agents by inhalation including dry powder inhalers (DPI),

metered-dose inhalers (MDI) and nebulizer inhalers. When preparing pharmaceutical compositions and formulations for use in such devices, it is highly desirable to have a crystalline form of the therapeutic agent that is neither hygroscopic nor deliquescent and which has a relatively high melting point thereby allowing the material to be micronized without significant decomposition. Although crystalline freebase forms of the compound of formula I have been reported in U.S. Patent Publication No. 2007/0112027 to Axt et al. as Form I and Form II, the crystalline freebase forms of the present invention have different and particularly useful properties, including higher melting points

One aspect of the invention relates to crystalline freebase forms of biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl]methy lamino } ethyl) piperidin-4-yl ester characterized by a powder x-ray diffraction pattern comprising diffraction peaks at 2Θ values of 6.6±0.1, 13.1±0.1, 18.6±0.1, 19.7±0.1, and 20.2±0.1.

Another aspect of the invention relates to a crystalline freebase of biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl]methy lamino } ethyl) piperidin-4-yl ester, designated as form III, which is characterized by a powder x-ray diffraction pattern comprising diffraction peaks at 2Θ values of 6.6±0.1, 13. l±O.l,

18.6±0.1, 19.7±0.1, and 20.2±0.1; and further characterized by having five or more additional diffraction peaks at 2Θ values selected from 8.8=1=0.1, 10. l±O.l, 11.4±0.1, l l.β±O.l, 14.8±0.1, 15.2±0.1, lβ.l±O.l, 16.4±0.1, 16.9±0.1, 17.5±0.1, 18.2±0.1, 19.3±0.1, 19.9±0.1, 20.8±0.1, 21. l±O.l, 21.7±0.1, and 22.3±0.1.

Still another aspect of the invention relates to a crystalline freebase of biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl]methy lamino } ethyl) piperidin-4-yl ester, designated as form IV, which is characterized by a powder x-ray diffraction pattern comprising diffraction peaks at 2Θ values of 6.6±0.1 , 13. l±O.1 ,

18.6=1=0.1, 19.7=1=0.1, and 20.2±0.1; and further characterized by having five or more additional diffraction peaks at 2Θ values selected from 10.6±0.1, 15.0=1=0.1, lβ.O±O.l, 17.3±0.1, 17.7±0.1, 20.9±0.1, 21.4±0.1, 22.6±0.1, 24.6±0.1, and 27.8±0.1.

Preparation 1

Biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l- ylmethvDbenzovHmethylaminol ethyDpiperidin-4-yl Ester The diphosphate salt of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (16 g) was dissolved in a biphasic mixture of water (100 mL) and EtOAc (200 mL). NaOH (2 N, 75 mL) was added over a period of 5 minutes. The mixture was then stirred for 30 minutes. The phases were separated and the aqueous phase was extracted with EtOAc (200 mL). The combined organic phases were concentrated. DCM (100 mL) was added, and the mixture evaporated to dryness. The solids were dried in an oven for about 48 hours to yield the title compound (9.6 g).

EXAMPLE 1

Crystalline Freebase of Biphenyl-2-ylcarbamic Acid l-(2-{r4-(4-Carbamoylpiperidin-l- ylmethyl)benzoyllmethylamino|ethyl)piperidin-4-yl Ester (Form III) Biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (102.4 mg) was dissolved in MeCN (500 μL). The solution was stirred at room temperature for 80 minutes and a white solid precipitate formed. The mixture was placed in the shaker block to thermocycle (0-40 0C in one hour blocks) for 48 hours. A white, dense, immobile solid was observed. MeCN (500 μL) was added to mobilize the slurry. The mixture was then placed back in the shaker block for 2 hours. The solids were isolated by vacuum filtration using a sinter funnel, then placed in the piston dryer at 40 0C under full vacuum for 15.5 hours, to yield 76.85 mg of the title crystalline compound.

EXAMPLE 2

Crystalline Freebase of Biphenyl-2-ylcarbamic Acid l-(2-{r4-(4-Carbamoylpiperidin-l- ylmethyl)benzoyllmethylamino|ethyl)piperidin-4-yl Ester (Form III) Diphosphate salt of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoyl-piperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (C3sH43NsO4»2H3PO4; MW 793.75; 632.9 g) was slurried in isopropyl acetate (11.08 L) and water (6.33 L) at room temperature under nitrogen. The suspension was warmed to 53±3 0C and 1OM NaOH (317 mL) was added to the stirred mixture, while maintaining the temperature of the mixture above 50 0C. The mixture was stirred for approximately 5 minutes at 53±3 0C before allowing the layers to settle. The layers were then separated and the aqueous layer was removed. Water (3.16 L) was added to the organic layer while maintaining the temperature of the mixture above 50 0C. The mixture was stirred for 5 minutes at 53±3 0C before allowing the layers to settle. The layers were separated and the water layer was removed. Isopropyl acetate (6.33 L) was added and then about 10 volumes of distillate were collected by atmospheric distillation. This step was repeated with additional isopropyl acetate (3.2 L). After the second distillation, the temperature of the clear solution was reduced to 53±3 0C, then seeded with a suspension of the biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester crystalline freebase (Form III; 3.2 g) in isopropyl acetate (51 mL). The resulting suspension was stirred at 53±3 0C for 2 hours, then cooled to 10±3 0C over 4 hours. The suspension was stirred at 10±3 0C for at least 2 hours and then the solids were collected by filtration. The resulting filter cake was washed with isopropyl acetate (2 x 1.9 L) and the product was dried in vacuo at 50 0C to yield the title crystalline compound (C3SH43NsO4; MW 597.76; 382.5 g, 80.3% yield).

EXAMPLE 3

Recrystallization of Crystalline Freebase of Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4- Carbamoylpiperidin- 1 -ylmethyDbenzoyllmethylaminol ethyl)piperidin-4-yl Ester (Form

III)

Crystalline freebase of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (Form III; C35H43N5O4; MW 597.76; 372.5 g) was slurried in toluene (5.6 L) at 20±3 0C under nitrogen. The suspension was warmed to 82±3 0C, and held at this temperature until complete dissolution was observed. The solution was then clarified into the crystallizer vessel, followed by rinsing with toluene (373 μL). Solids were observed in the crystallizer vessel, and the vessel was re-heated to 82±3 0C to effect dissolution, then cooled to 58±3 0C and seeded with a pre-sonicated (approximately 1 minute) of crystalline freebase (Form III; 1.9 g) in toluene (8 μL). The resulting suspension was allowed to stand at 58±3 0C for at least 4 hours, then cooled to 20±3 0C over 2 hours (approximate cooling rate of 0.33 °C/min). The suspension was stirred at 20±3 0C for at least 1 hour, then the solids were collected by filtration. The resulting filter cake was washed with toluene (2 x 1.2 L) and the product was dried in vacuo at 52±3 0C to yield the title crystalline compound (345.3 g, 92.7% yield).

EXAMPLE 4

Crystalline Freebase of Biphenyl-2-ylcarbamic Acid l-(2-{r4-(4-Carbamoylpiperidin-l- ylmethyl)benzoyllmethylamino|ethyl)piperidin-4-yl Ester (Form IV) Biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester (prepared as described in Preparation 1; 2.5 g) was dissolved in MeCN (10 mL) to yield a viscous oily pale yellow material. Additional MeCN (5 mL) was added to dilute the material. The solution was seeded with crystalline freebase (20 mg; Form III prepared as described in Example 1) and stirred at room temperature for 90 minutes. A large amount of white precipitate (small crystals) was observed. The slurry was analyzed under a polarized light microscope and found to be birefringent.

Additional MeCN (3 mL) was added and the slurry was placed in a Metz SynlO block to thermocycle (0-40 0C in one hour blocks) at 800 rpm overnight. The Metz SynlO is a 10 position parallel reaction station that is static. Agitation of the solution/slurry was by a cross magnetic stirrer bar. The shaker block was a separate piece of equipment that was heated and cooled by an external Julabo bath. The material was removed at 0 0C. It was observed that the slurry had settled out, leaving a pale yellow solution above the white precipitate. The slurry was stirred and placed back in the shaker block to thermocycle.

The material was removed at 40 0C, and stirred at a high agitation rate at room temperature for 80 minutes. The slurry was again analyzed and found to be birefringent. The filter cake was isolated by vacuum filtration using a sinter funnel. MeCN (3 mL) was used to wet the filter paper and the filter cake was washed with MeCN prior to filtration. The cake was deliquored under vacuum for 40 minutes to yield 2.3 g of a flowing white powder. The material was placed in a piston dryer at 400C for 65 hours, to yield 2.2 g of the title crystalline compound as a white powder (99.6% purity).

PATENT

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=0049F6A3F9FB8C7273B825D49F2465F6.wapp1nA?docId=WO2005087738&tab=PCTDESCRIPTION&maxRec=1000

Example 1
Biphenyl-2-ylcarbamic Acid l-(2-{[4-(4-Carbamoylpiperidin-l- ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl Ester

To a three-necked 2-L flask was added isonipecotamide (5.99 g, 40.0 mmol), acetic acid (2.57 mL), sodium sulfate (6.44 g) and LPA (400 mL). The reaction mixture was cooled to 0-10°C with an ice bath and a solution ofthe product of Preparation 5 (11 g, 22.7 mmol) in LPA (300 mL) was slowly added. The reaction mixture was stined at room temperature for 2 hours and then cooled to 0-10°C. Sodium triacetoxyborohydride (15.16 g, 68.5 mmol) was added portion wise and this mixture was stined at room temperature for 16 h. The reaction mixture was then concentrated under reduced pressure to a volume of about 50 mL and this mixture was acidified with IN HCl (200 mL) to pH 3. The resulting mixture was stined at room temperature for 1 hour and then extracted with DCM (3 x 250 mL). The aqueous phase was then cooled to 0-5°C with an ice bath and 50% aqueous NaOH solution was added to adjust the pH ofthe mixture to 10. This mixture was then extracted with isopropyl acetate (3 x 300 mL) and the combined organic layers were washed with water (100 mL), brine (2 x 50 mL), dried over sodium sulfate, filtered and concentrated to afford 10.8 g ofthe title compound (80% yield. MS m/z: [M + H“1”] calcd for C35H43N5O4, 598.3; found, 598.6. Rf = 2.32 min (10-70 ACN: H2O, reverse phase HPLC).

Example 1A
Biphenyl-2-ylcarbamic acid l-(2- {[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl] methylamino} ethyl)piperidin-4-yl ester was also prepared as a diphosphate salt using the following procedure :
5.0 g ofthe product of Example 1 was combined with 80 ml of IPA:ACN (1:1). 4.0 ml of water was added and the mixture heated to 50°C under stining, forming a clear solution. To this was added dropwise at 50°C, 16 ml 1M phosphoric acid. The resulting cloudy solution was stined at 50°C for 5 hours, then allowed to cool to ambient temperature, under slow stirring, overnight. The resulting crystals were collected by filtration and air-dried for 1 hour, then under vacuum for 18 hours, to give the diphosphate salt ofthe title compound (5.8 g, 75% yield) as a white crystalline solid (98.3% purity by HPLC).

Example IB
Biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl] methylamino }ethyl)piperidin-4-yl ester was also prepared as a monosulfate salt using the following procedure.
442 mg ofthe product of Example 1 (0.739 mmol of 96% pure material) was taken up in 5 ml of H2O:ACN (1:1) and 1.45 ml of IN sulfuric acid was added slowly, while monitoring the pH. The pH was adjusted to approx. pH 3.3. The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness. 161 g of the lyophilized material was dissolved in 8.77 ml of IPA:ACN (10:1). The suspension was heated by placing the vial in a pre-heated 70°C water bath for 1.5 hours. Oil droplets formed within 5 minutes. The heat was lowered to 60°C and the mixture heated for an additional 1.5 hours, followed by heating at 50°C for 40 minutes, at 40°C for 40 minutes, then at 30°C for 45 minutes. The heat was turned off and the mixture was allowed to slowly cool to room temperature. The next day, the material was viewed under a microscope and indicated needles and plates. The material was then heated at 40°C for 2 hours, at 35°C for 30 minutes, and then at 30°C for 30 minutes. The heat was turned off and the mixture was allowed to slowly cool to room temperature. The solid was then filtered and dried using a vacuum pump for 1 hour to give the monosulfate salt ofthe title compound (117 mg, 73% yield).

Example IC
Biphenyl-2-ylcarbamic acid l-(2- {[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl] methylamino} ethyl)piperidin-4-yl ester was also prepared as a dioxalate salt using the following procedure.
510 mg ofthe product of Example 1 (0.853 mmol of 96% pure material) was taken up in 5 ml of H2O:ACN (1:1) and 1.7 ml of 1M aqueous oxalic acid was added slowly, while monitoring the pH. The pH was adjusted to approx. pH 3.0. The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness. 150 mg ofthe lyophilized material was dissolved in 13.1 ml of 94%IPA/6%H20. The mixture was stined in a pre-heated 60°C water bath for 2.5 hours. The heat was turned off and the mixture was allowed to cool to room temperature. The vial was refrigerated at 4°C. After 6 days, an oily material was observed with what appeared to be a crystal on the side ofthe vial. The vial was then allowed to reach room temperature, at which point seeds (synthesis described below) were added and allowed to sit for 16 days. During this time, more crystals were observed to come out of solution. The solid was then filtered and dried using a vacuum pump for 14 hours to give the dioxalate salt ofthe title compound (105 mg, 70% yield).
Seed Synthesis
510 mg ofthe product of Example 1 (0.853 mmol of 96% pure material) was taken up in 5 ml of H2O:ACN (1:1) and 1.7 ml of 1M aqueous oxalic acid was added slowly, while monitoring the pH. The pH was adjusted to approx. pH 3.0. The clear solution was filtered through a 0.2 micron filter, frozen and lyophilized to dryness to yield a dioxalate salt. 31.5 mg of this dioxalate salt was dissolved in 2.76 ml of 94%IPA/6%H20. The mixture was stined in a pre-heated 60°C water bath for 2.5 hours. After 25 minutes, all of the sample was in solution. The heat was turned off and the mixture was allowed to cool to room temperature. The next day, a small amount of viscous material was present. The vial was refrigerated at 4°C. After 4 days, the viscous material was still present. The vial was then placed at room temperature and observed one month later. The material appeared to be solid, and was observed to be crystalline under a microscope. The solid was then » filtered and dried using a vacuum pump for 1 hour to give the dioxalate salt (20 mg, 63.5% yield).

Example ID
Biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl] methylamino} ethyl)piperidin-4-yl ester was also prepared as a freebase crystal using the following procedure.
230 mg ofthe product of Example 1 was dissolved in 0.2 ml of H O:ACN (1:1), using slight heat. The mixture was then heated in a 70°C water bath for 2 hours. The heat was turned off and the mixture was allowed to cool to room temperature, then refrigerated at 4°C for 1 hour. 50 μl of water was then added (oiled out), followed by the addition of 40 μl of ACN to get the sample back into solution. Seeds (synthesis described below) were added under slow stirring at room temperature. Crystals started to form ,and the mixture was allowed to sit overnight, with slow stirring. The next day, a heat cool cycle was applied (30°C for 10 minutes, 40°C for 10 minutes, then 50°C for 20 minutes). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, a second heat/cool cycle was applied (60°C for 1 hour, with dissolving observed at 70°C). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, crystals were present and a third heat cool cycle was applied (60°C for 3 hours). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. The next day, a heat cool cycle was applied (60°C for 3 hours, slow cool, then 60°C for 3 hours). The heat was turned off and the mixture allowed to cool overnight, with slow stirring. After 3 days, the solid was filtered and placed on a high vacuum line to remove all solvent and give a freebase crystal ofthe title compound.

Seed Synthesis
109 mg ofthe product of Example 1 was dissolved in 0.56 ml of H2O:ACN (1:1). The suspension was left in a vial (cap loosely placed on top) to allow for a slower evaporation time. The vial was placed under a nitrogen flow environment, although the nitrogen was not used for evaporation, only for the environment. A precipitate was visible within 1 day, which was observed to be crystalline under a microscope. The solid was then placed on a high vacuum line to remove all solvent to give the freebase crystal.
Quantitative recovery, 97.8% pure by HPLC.

Example IE
Biphenyl-2-ylcarbamic acid 1 -(2- { [4-(4-carbamoylpiperidin- 1 -ylmethyl)benzoyl] methylamino} ethyl)piperidin-4-yl ester was also prepared as a freebase crystal using the following alternate procedure.
70 mg ofthe product of Example 1 was dissolved in 0.1 mL ACN. After addition of 0.3 ml MTBE, the solution appeared cloudy. An additional 50 μl of ACN was added to clarify the solution (155 mg/ml ACNMTBE = 1 :2). The mixture was left in the vial and capped. A solid appeared by the next day. The solid was then filtered and placed on a high vacuum line to remove all solvent and give a freebase crystal ofthe title compound.

PATENT

https://patents.google.com/patent/WO2012009166A1/en

U.S. Patent No. 7,228,657 to Mammen et al. discloses novel biphenyl compounds that are expected to be useful for treating pulmonary disorders such as chronic obstructive pulmonary disease and asthma. In particular, the compound biphenyl-2-ylcarbamic acid 1- (2- {[4-(4-carbamoylpiperidin-l-ylmethyl)benzoyl]methylamino}-ethyl)piperidin-4-yl ester is specifically described in this application as possessing muscarinic receptor antagonist or anticholiner ic activity, and is represented by formula I:

Figure imgf000002_0001

The compound of formula I is synthesized from the compound 8, which is described as being prepared from the oxidation of 2-(benzylmethylamino)ethanol to the aldehyde intermediate followed by reductive amination with biphenyl-2-yl-carbamic acid piperidin- 4-yl ester and debenzylation:

Figure imgf000003_0001
Figure imgf000003_0002

However, while this procedure performs well on small scale, the aldehyde intermediate is difficult to scale up due to its instability, and low yields were typically observed.

Thus, a need exists for an efficient process of preparing compound 8 as a pure material with high chemical purity and good overall yield, without having to isolate intermediates. This invention addresses those needs.

Therapeutic agents useful for treating pulmonary or respiratory disorders are advantageously administered directly into the respiratory tract by inhalation. In this regard, several types of pharmaceutical inhalation devices have been developed for administering therapeutic agents by inhalation including dry powder inhalers, metered- dose inhalers, and nebulizer inhalers. When preparing pharmaceutical compositions and formulations for use in such devices, it is highly desirable to have a crystalline form of the therapeutic agent that is neither hygroscopic nor deliquescent and which has a relatively high melting point thereby allowing the material to be micronized without significant decomposition.

A crystalline diphosphate of the compound of formula I has been reported in U.S. Patent No. 7,700,777 to Axt et al, and a crystalline freebase (identified as Form III) is described in U.S. Patent Application Publication No. 201 1/0015163 to Woollham. All of the aforementioned disclosures are incorporated herein by reference.

The compound of formula I is described as being prepared by reacting compound 8 with 4-carboxybenzaldehyde to form the aldehyde core 10:

Figure imgf000004_0001

which is then isolated prior to being combined with isonipicotamide in the presence of a reducing agent to form the compound of formula I. The crystalline diphosphate is prepared by contacting the separated and purified compound of formula I with phosphoric acid. The crystalline freebase (Form III) can then be prepared from the crystalline diphosphate.

A need also exists for an efficient process of preparing the crystalline freebase (Form III). It is desirable to develop a process that does not first require preparation of the crystalline diphosphate. This invention addresses those needs.

Figure imgf000011_0001
Figure imgf000013_0001
Figure imgf000014_0001

Preparation 1

Biphenyl-2-yl-carbamic acid piperidin-4-yl Ester

Figure imgf000018_0001

Biphenyl-2-isocyanate (97.5 g, 521 mmol) and 1 -benzylpiperidin-4-ol (105 g, 549 mmol) were heated together at 70°C for 12 hours. The mixture was then cooled to 50°C and EtOH (1 L) was added, followed by the slow addition of 6M HC1 (191 mL). The resulting mixture was then cooled to ambient temperature. Ammonium formate (98.5 g, 1.6 mol) was added and then nitrogen gas was bubbled through the solution vigorously for 20 minutes. Palladium on activated carbon (20 g, 10 wt% dry basis) was added and the mixture was heated at 40°C for 12 hours, and then filtered. The solvent was removed under reduced pressure and 1M HC1 (40 mL) was added to the crude residue. The pH of the mixture was adjusted with 10 N NaOH to pH 12. The aqueous layer was extracted with EtOAc (2×150 mL), and the organic layer was dried over MgS04, filtered and the solvent removed under reduced pressure to yield the title compound (155 g). HPLC (10-70) ¾ = 2.52; m/z: [M + H+] calcd for Ci8H2202 297.15; found 297.3.

EXAMPLE 1

Step A: (2,2-Dimethoxyethyl)methylcarbamic Acid Benzyl Ester

Figure imgf000018_0002

K2CO3 (13.8 g, 100 mmol, 1.76 eq.) and H20 (46 mL) were mixed to form a homogeneous solution. The solution was cooled to 20°C. N-methylaminoacetaldehyde dimethylacetal (12.8 mL, 100 mmol, 1.8 eq) and MeTHF (50 mL) were added. The resulting mixture was cooled to 2°C. Benzyl chloroformate (8.1 mL, 56.7 mmol, 1.0 eq.) was added by syringe over 10 minutes (addition was exothermic). The mixture was maintained at room temperature until completion of the reaction. The layers were separated and the organic layer was washed with IN HC1 (50 mL) and used directly in the next step.

Step B: Methyl-(2-oxoethyl)carbamic Acid Benzyl Ester

Figure imgf000019_0001

The mixture from the previous step was combined with a 3N HC1 solution (70 mL), and the resulting mixture was stirred for 18 hours at 22°C to yield a clear homogeneous pale yellow solution. Solid aHC03 was added to the solution to bring the pH to neutral. The layers were separated and the aqueous layer was back-extracted with MeTHF (20 mL). The organic layers were combined and washed with a saturated aHC03 solution (50 mL). The layers were separated and the organic layer was dried over Na2S04, filtered and concentrated to dryness to afford the title compound (1 1.9 g) as a pale yellow oil.

Step C: Biphenyl-2-yl-carbamic acid l-[2-(benzyloxycarbonyl

methylamino)ethyl]piperidin-4-yl Ester

Figure imgf000019_0002

Biphenyl-2-yl-carbamic acid piperidin-4-yl ester (31.1 g, 105 mmol, 1.0 eq.) and MeTHF (150 mL) were mixed. A solution of methyl-(2-oxoethyl)carbamic acid benzyl ester (23 g, 113.4 mmol, 1.05 eq.) in MeTHF (150 mL) was prepared and added to the ester mixture. The resulting mixture was heated to 30°C for a few minutes, then cooled to room temperature over 1 hour. The mixture was then cooled to 3°C and the temperature maintained for 1 hour. NaHB(OAc)3 (35.1 g, 170 mmol, 2.0 eq.) was added portion-wise while maintaining the internal temperature at 7±1°C. After addition, the mixture was allowed to warm to room temperature until the reaction was complete. A saturated solution of aHC03 (3000 mL) was added, stirred for 20 minutes, and the layers separated. This was repeated, after which the organic layer was dried over a2S04. The material was filtered, concentrated and dried under high vacuum to afford the title compound (43 g) as a thick colorless to pale yellow oil, which was used directly in the next step without purification.

Step D: Biphenyl-2-yl-carbamic acid l-(2-methylaminoethyl)piperidin-4-yl Ester

Figure imgf000020_0001

Biphenyl-2-yl-carbamic acid l-[2-(benzyloxycarbonyl methylamino)ethyl] piperidin-4-yl ester (53 g, 105 mmol, 1 eq.), MeOH (250 mL), and MeTHF (50 mL) were combined under nitrogen. 10% palladium on carbon (0.8 g) was added and hydrogen was bubbled into the mixture for 1 minute. The reaction vessel was sealed and stirred under hydrogen at atmospheric pressure for three hours. The mixture was then filtered, and the solids were washed MeTHF (10 mL).

The filtrate and washes were combined and concentrated under reduced pressure (250 mL removed). MTBE (100 mL) was added, and the solution again concentrated under reduced pressure (100 mL removed). MTBE (200 mL) was added and the solution was seeded with a few milligrams of biphenyl-2-yl-carbamic acid l-(2-methylaminoethyl) piperidin-4-yl ester, and the mixture was maintained for 3 hours. The solids were collected and the vessel and filter cake were washed with MTBE (2×15 mL). The material was dried to yield 13.2 g of the title compound (99.5% pure). This process was repeated to yield the title compound (12.5 g, 98.6% pure). The filtrate and washes were combined and concentrated under reduced pressure. MTBE (150 mL) was added and the solution was seeded with a few milligrams of biphenyl-2-yl-carbamic acid l-(2-methylaminoethyl) piperidin-4-yl ester, and the mixture was maintained for 20 hours. The solids were collected and the vessel and filter cake were washed with MTBE (2×15 mL). The material was dried to yield the title compound (5 g, 90% pure).

A portion of the three crops (13 g , 12 g, 4.5 g, respectively) were combined taken up in IPA (90 mL). The resulting slurry was heated to 45°C, then cooled to room temperature over 1 hour. The slurry was stirred for 5 hours at 25°C. The solids were collected and washed with IPA (2×15 mL). The solids were then dried for 1 hour to yield the title compound (25 g, >99% pure).

EXAMPLE 2

All volumes and molar equivalents are given relative to biphenyl-2-yl-carbamic acid piperidin-4-yl ester.

Step A: (2,2-Dimethoxyethyl)methylcarbamic Acid Benzyl Ester K2C03 (8.4 kg, 60 mol, 1.8 eq.) and H20 (49.3 kg, 2.6 volumes) were placed in the reaction vessel and stirred. N-methylaminoacetaldehyde dimethylacetal (6.5 kg, 54 mol, 1.6 eq) and MeTHF (20.2 kg, 2.9 volumes) were added. The resulting mixture was cooled to 5°C. Benzyl chloroformate (6.8 kg, 37.6 mol, 1.1 eq.) was added over a period of about 30 minutes, while maintaining the temperature below 10°C. The feed line was rinsed with MeTHF (4.3 kg). The mixture was then maintained at 5°C and stirred for 1 hour. The layers were separated and the organic layer was washed with IN HC1 (14.3 kg, 1 1.7 mol, 1.4 volumes) and used directly in the next step.

Step B: Methyl-(2-oxoethyl)carbamic Acid Benzyl Ester

The mixture from the previous step was combined with water (23.4 kg,

2.9 volumes) and 30% hydrochloric acid (13.1 kg, 107.7 mol, 1.1 volumes). Water (5.1 kg) was used to rinse the feed line. The temperature was adjusted to 25-30°C, and the reaction was run for 16-24 hours. A 25% NaOH solution (1 1.8 kg, 71.1 mol, 2.2 eq.) was added to the solution to adjust the pH and obtain phase separation.

The layers were separated and the aqueous layer was back-extracted with MeTHF

(10.0 kg, 1.1 volumes). The aqueous layer was discarded and the organic layers were combined. MeTHF (4.4 kg) was used to rinse the feed line. The organics were washed with a saturated aHC03 solution (14.6 kg, 15.6 mol, 1.1 volumes). The layers were separated and the organic layer was dried over a2S04 (2.5 kg, 17.6 mol) for 60-90 minutes. The drying agent was filtered off and the remaining solids were washed with

MeTHF (8.8 kg, 1 volume). The reaction vessel was washed with water and MeOH before continuing with the next step.

Step C: Biphenyl-2-yl-carbamic acid l-[2-(benzyloxycarbonyl

methylamino) ethyl Jpiperidin-4-yl Ester

The product from the previous step (in MeTHF) and biphenyl-2-yl-carbamic acid piperidin-4-yl ester (10.0 kg, 32.6 mol, 1.0 eq.) in MeTHF (28.5 kg) were placed in the reaction vessel and heated to 30°C for one hour. The mixture was then cooled to 5°C. NaHB(OAc)3 (10.0 kg, 45.8 mol, 1.4 eq.) was added portion wise over a period of 40 minutes while maintaining the temperature below 20°C. The mixture was then stirred for 30 minutes. Additional NaHB(OAc)3 (0.5 kg) was added the reaction allowed to progress to completion. A saturated solution of NaHCC^ (14.3 kg, 15.3 mol, 1.1 volumes) was added and stirred for 10 minutes. The aqueous phase was separated and discarded. A 33% NaOH solution (15.8 kg, 129.9 mol, 4.0 eq.) was added to the reaction mixture to adjust the H to be in the range of 8-12. Water (40 kg) was added in two portions, after which phase separation occurred. A saturated NaHCC (7.1 kg, 7.6 mol, 0.7 volumes) was added to the reaction mixture and stirred for 10 minutes. The aqueous phase was separated and discarded. Additional water (4.9 kg) was added to dissolve any remaining salts and a vacuum distillation was conducted at a maximum temperature of 45°C to remove part of the solvent (7.2 volumes). MeOH (56.1 kg, 7.2 volumes) was added to the reaction mixture before continuing with the next step.

Step D: Biphenyl-2-yl-carbamic acid l-(2-methylaminoethyl)piperidin-4-yl Ester

10% palladium on carbon (0.4 kg, 0.03 wt%, Degussa type 101 NE/W) was added to the reaction mixture. A hydrogenation reaction was performed to remove the benzyloxycarbonyl protective group, with reaction conditions at 30±5°C and 4 bar pressure. The reaction was run until completion. The mixture was then filtered and the filter cake was washed with MeOH (8.0 kg, 1.0 volume). The reaction was continued in a clean vessel, which was charged with the product solution (in MeTHF/MeOH) from the hydrogenation reaction. 3-Mercaptopropyl silica (0.6 kg, 0.07 wt%, Silicycle) was added. MeOH (4.8 kg) was used to rinse the feed line. The reaction mixture was stirred for 14-72 hours at 25±5°C. Activated carbon (0.7 kg, 0.07 wt%) was added and the mixture stirred for 30 minutes. The mixture was filtered and the filter cake was washed with MeOH (1.0 volume). The reaction was continued in a clean vessel, which was charged with the product solution (in MeTHF/MeOH), and MeOH (4.2 kg) was used to rinse the feed line. The mixture was heated to 40-45°C and a vacuum distillation was performed to bring the final volume to 5.6 volumes (removal of methanol).

2-propanol (40.2 kg, 5.0 volumes) was added and distillation continued until the volume was reduced to 2.5 volumes. The solids were then isolated by filtration and washed with MTBE (1.5 volumes) to yield the product as a wet cake (8.6 kg, 96.8% purity). The cake was charged to the reaction vessel and additional 2-propanol

(1.9 volumes) was added. The mixture was warmed to 40±5°C, and maintained at that temperature for 2 hours. The mixture was then slowly cooled over a minimum of 4 hours to 20°C, then actively cooled to 5-10°C, followed by stirring for 2 hours. The product was filtered and the resulting cake washed with MTBE (1.0 volume). The solids were then dried under atmospheric conditions to yield the title compound (6.6 kg, 98.5% purity).

EXAMPLE 3

Crystalline Freebase of Biphenyl-2-yl-carbamic Acid l- {2-r(4-carbamoylbenzoyl) methylaminolethyllpiperidin-4-yl Ester (Form III)

Biphenyl-2-yl-carbamic acid l-{2-[(4-formylbenzoyl)

methylamino ] ethyl }piperidin-4-yl Ester

Figure imgf000023_0001

4-Carboxybenzaldehyde (9 g, 60 mmol, 1.0 eq.) and biphenyl-2-yl-carbamic acid 1-

(2-methylaminoethyl)piperidin-4-yl ester (21.2 g, 60 mmol, 1.0 eq.) were combined in MeTHF (115 mL). The mixture was stirred for 0.5 hours, forming a thick slurry.

Additional MeTHF (50 mL) was added to form a free-flowing slurry. 4-(4,6-dimethoxy- l,3,5-triazin-2-yl)-4-methylmorpholinium chloride (18 g, 63 mmol, 1.1 eq., 97% pure) was added in two portions and the funnel rinsed with additional MeTHF (50 mL). The mixture was stirred at room temperature overnight. MeCN (50 mL) was added and the mixture was filtered. The solids were washed with MeTHF (30 mL). The filtrate and washes were combined and a saturated aHC03 solution (100 mL) was added and stirred for 10 minutes. The layers were separated and a saturated NaCl solution (100 mL) was added and stirred for 10 minutes. The layers were separated and the aqueous layer discarded. The resulting solution was concentrated under reduced pressure and held at room temperature for three days, then used directly in the next step.

Step B: Biphenyl-2-yl-carbamic acid l-{2-[(4-carbamoylbenzoyl)

meth lamino] ethyl}piperidin-4-yl ester (non-isolated form)

Figure imgf000023_0002

Isonipecotamide (15.4, 120 mmol, 2.0 eq.) and IPA (200 mL) were added to the solution of biphenyl-2-yl-carbamic acid l-{2-[(4-formylbenzoyl)methylamino]ethyl} piperidin-4-yl ester from the previous step. Liquid (200 mL) was distilled off and additional IPA (400 mL) was added under reduced pressure at 60°C. Liquid (400 mL) was distilled off over a period of 1.5 hours and additional IPA (600 mL) was added. Liquid (100 mL) was distilled off and the remaining solution was cooled to 30°C to yield a hazy white mixture, which was then added to Na2S04 (18 g). The flask was rinsed with IPA (100 mL) and added to the solution. The resulting mixture was cooled to room

temperature and AcOH (20 mL, 360 mmol, 6.0 eq.) was added. The mixture was cooled to 18°C with an ice bath and NaHB(OAc)3 (38.2 g, 180 mmol, 3.0 eq.) was added over 5 minutes. The mixture was allowed to warm up to 25°C and was maintained at that temperature for 2 hours. Solvent was removed under reduced pressure, and the remaining material was used directly in the next step.

Step C: Biphenyl-2-yl-carbamic acid l-{2-[(4-carbamoylbenzoyl)

methylamino]ethyl}piperidin-4-yl ester (isolated solid)

iPrOAc (300 mL) was added to the material, followed by the addition of water (200 mL). The pH of the solution was adjusted to pH 1 with 3N HC1 (-150 mL). The layers were separated and the organic layer was discarded. The aqueous layer was collected, and iPrOAc (300 mL) was added. The pH of the solution was adjusted to basic pH with 50 wt% NaOH (-100 mL). The resulting mixture was stirred for 15 minutes and the layers were separated. The organic layer was filtered and seeded with micronized crystalline freebase of biphenyl-2-yl-carbamic acid l- {2-[(4-carbamoylbenzoyl) methylamino]ethyl}piperidin-4-yl ester (Form III; prepared as described in U.S. Patent Application Publication No. 201 1/0015163 to Woollham) and stirred overnight at room temperature to yield a white slurry. Stirring was continued for 8 hours at room temperature and for 16 hours at 5°C (cold room). The mixture was slowly filtered under pressure. The cake was washed with cold iPrOAc (2×20 mL) and dried under nitrogen to yield a white solid (27.5 g). The material was further dried in a vacuum oven at 30°C for 24 hours to yield 25.9 g.

Step D: Crystalline Freebase of Biphenyl-2-yl-carbamic Acid l-{2-[ ( 4- carbamoylbenzoyl)methylamino]ethyl}piperidin-4-yl Ester (Form III) The white solid (5 g, 60 mmol, 1.0 eq.) was dissolved in toluene (75 mL) and the resulting mixture was heated to 82°C to yield a clear solution. The solution was filtered. The solids were washed with toluene (2 x 5 mL), and the filtrate and washes were combined. The mixture was cooled to 60°C and seeded with micronized crystalline freebase of biphenyl-2-yl-carbamic acid l-{2-[(4-carbamoylbenzoyl)methylamino]ethyl} piperidin-4-yl ester (Form III; prepared as described in Example 3 in U.S. Patent

Application Publication No. 201 1/0015163 to Woollham). The mixture was maintained at 55°C for 2 hours, then cooled to room temperature on an oil bath overnight (~16 hours). The resulting slurry was then filtered and the cake was dried for 3 hours to yield a solid while material (4.6 g). The material was further dried in a vacuum oven at 30°C for 24 hours (exhibited no further weight loss) to yield the title compound (4.6 g).

The product was analyzed by powder x-ray diffraction, differential scanning calorimetry and thermal gravimetric analysis, and was determined to be the crystalline freebase (Form III) of biphenyl-2-ylcarbamic acid l-(2-{[4-(4-carbamoylpiperidin-l- ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester described in U.S. Patent Application Publication No. 201 1/0015163 to Woollham.

US20050113417A1 *2003-11-212005-05-26Mathai MammenCompounds having beta2 adrenergic receptor agonist and muscarinic receptor antagonist activity
WO2006099165A1 *2005-03-102006-09-21Theravance, Inc.Crystalline forms of a biphenyl compound
US7228657B22003-07-102007-06-12Controlled Environments LimitedClimate control for a greenhouse
US20110015163A12009-07-152011-01-20Grahame WoollamCrystalline freebase forms of a biphenyl compound
Family To Family Citations
JP4555283B2 *2003-02-142010-09-29セラヴァンス, インコーポレーテッドβ2 adrenergic receptor agonist activity and biphenyl derivatives having muscarinic receptor antagonist activity
CN1930125B *2004-03-112010-07-21施万制药Biphenyl compounds useful as muscarinic receptor antagonists
US7659403B2 *2005-03-102010-02-09Theravance, Inc.Biphenyl compounds useful as muscarinic receptor antagonists
Patent ID

Title

Submitted Date

Granted Date

US9226896 CRYSTALLINE FREEBASE FORMS OF A BIPHENYL COMPOUND
2014-11-19
2015-06-18
US9656993 CRYSTALLINE FORMS OF A BIPHENYL COMPOUND
2015-12-18
2016-06-16
US7700777 Crystalline forms of a biphenyl compound
2007-12-27
2010-04-20
Patent ID

Title

Submitted Date

Granted Date

US9415041 Crystalline freebase forms of a biphenyl compound
2015-12-01
2016-08-16
US9249099 CRYSTALLINE FORMS OF A BIPHENYL COMPOUND
2014-11-25
2015-06-04
US8921396 Crystalline freebase forms of a biphenyl compound
2013-08-22
2014-12-30
US7521041 Biphenyl compounds useful as muscarinic receptor antagonists
2008-04-24
2009-04-21
US2007112027 Crystalline forms of a biphenyl compound
2007-05-17
Patent ID

Title

Submitted Date

Granted Date

US8017783 Biphenyl compounds useful as muscarinic receptor antagonists
2008-03-20
2011-09-13
US7550595 Biphenyl compounds useful as muscarinic receptor antagonists
2007-12-20
2009-06-23
US9283183 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2014-11-12
2015-06-18
US2010048622 CRYSTALLINE FORMS OF A BIPHENYL COMPOUND
2010-02-25
US9452161 Biphenyl compounds useful as muscarinic receptor antagonists
2016-02-05
2016-09-27
Patent ID

Title

Submitted Date

Granted Date

US8754225 PROCESS FOR PREPARING A BIPHENYL-2-YLCARBAMIC ACID
2012-01-19
US8921395 Crystalline forms of a biphenyl compound
2014-03-19
2014-12-30
US8716313 Crystalline forms of a biphenyl compound
2013-01-14
2014-05-06
US8557997 Biphenyl compounds useful as muscarinic receptor antagonists
2012-08-23
2013-10-15
US8541451 CRYSTALLINE FREEBASE FORMS OF A BIPHENYL COMPOUND
2011-01-20
Patent ID

Title

Submitted Date

Granted Date

US8377965 CRYSTALLINE FORMS OF A BIPHENYL COMPOUND
2010-10-07
US8242137 CRYSTALLINE FORMS OF A BIPHENYL COMPOUND
2010-01-28
2012-08-14
US2017204061 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2016-08-30
US9765028 CRYSTALLINE FREEBASE FORMS OF A BIPHENYL COMPOUND
2016-07-11
US9035061 PROCESS FOR PREPARING A BIPHENYL-2-YLCARBAMIC ACID
2013-11-26
2014-05-01
Patent ID

Title

Submitted Date

Granted Date

US7803812 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2009-09-10
2010-09-28
US7910608 Biphenyl compounds useful as muscarinic receptor antagonists
2009-01-15
2011-03-22
US7491736 Biphenyl compounds useful as muscarinic receptor antagonists
2007-12-20
2009-02-17
US7585879 Biphenyl compounds useful as muscarinic receptor antagonists
2007-11-15
2009-09-08
US7288657 Biphenyl compounds useful as muscarinic receptor antagonists
2005-09-15
2007-10-30
Patent ID

Title

Submitted Date

Granted Date

US8912334 Biphenyl compounds useful as muscarinic receptor antagonists
2013-09-11
2014-12-16
US8273894 Biphenyl compounds useful as muscarinic receptor antagonists
2012-04-03
2012-09-25
US8173815 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2011-12-29
2012-05-08
US8053448 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2011-06-02
2011-11-08
US8034946 BIPHENYL COMPOUNDS USEFUL AS MUSCARINIC RECEPTOR ANTAGONISTS
2010-09-30
2011-10-11

/////////TD-4208, UNII:G2AE2VE07O, ревефенацин ريفيفيناسين 瑞维那新 , GSK 1160724, revefenacin, PHASE 3

CN(CCN1CCC(CC1)OC(=O)NC2=CC=CC=C2C3=CC=CC=C3)C(=O)C4=CC=C(C=C4)CN5CCC(CC5)C(=O)N

Sarecycline , サレサイクリン


Sarecycline.svg

ChemSpider 2D Image | Sarecycline | C24H29N3O8

Sarecycline

サレサイクリン

MW 487.5024, MF C24H29N3O8 FREE FORM

Paratek  INNOVATOR

FDA 2018/10/1 APPROVED SEYSARA, ALMIRALL, for the oral treatment of inflammatory lesions of non-nodular moderate to severe acne vulgaris in patients 9 years of age and older

(4S,4aS,5aR,12aS)-4-(dimethylamino)-3,10,12,12a-tetrahydroxy-7-[(methoxymethylamino)methyl]-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide
(4S,4aS,5aR,12aS)-4-(Dimethylamino)-3,10,12,12a-tetrahydroxy-7-{[methoxy(methyl)amino]methyl}-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydro-2-tetracenecarboxamide
1035654-66-0 [RN] FREE FORM
2-Naphthacenecarboxamide, 4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-7-[(methoxymethylamino)methyl]-1,11-dioxo-, (4S,4aS,5aR,12aS)-
94O110CX2E
9743

P005672, 

  • P 005672

Sarecycline hydrochloride.png

CAS 1035979-44-2 HCl

Molecular Formula C24 H29 N3 O8 . Cl H
 Molecular Weight 523.963

P-005672
PTK-AR-01
SC-1401
WC-3035

Sarecycline (trade name Seysara; development code WC-3035) is a tetracycline-derived antibiotic. In the United States, it was approved by the FDA in October 2018 for the treatment of moderate to severe acne vulgaris.[1]

Paratek Pharmaceuticals, Inc. licensed the US rights to sarecycline for the treatment of acne in the United States to Actavis, a subsidiary of Allergan, while retaining rights in the rest of the world.[2]

Allergan initiated a Phase 3 study in December 2014 evaluating the efficacy and safety of sarecycline tablets 1.5 mg/kg per day taken orally for 12 weeks versus placebo in the treatment of acne vulgaris.[3] Two phase 3 randomized, multi-center, double-blind, placebo-controlled studies evaluating the efficacy and safety of sarecycline in moderate to severe acne reported positive results on 27 March 2017.[4]

SYN

US 2016/0200671

PATENT

WO 2008079363

PATENT

WO 2008079339

PATENT

WO 2012155146

EXAMPLES

[00104] The following examples illustrate the synthesis of the compounds described herein.

Synthesis of (4S,4aS,5aR,12aS)-4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid amide (“the free base”).

[00105] A solution of 7-formylsancycline TFA salt (2.23 g) and N,0-dimethylhydroxylamine hydrochloride (780 mg) in N,N-dimethylacetamide (15 mL) was stirred for 10 minutes at room temperature under argon atmosphere. To this solution was added sodium cyanoborohydride (302 mg). The solution was stirred for 5 minutes and monitored by LC-MS. The reaction mixture was poured into diethyl ether, and the resulting precipitates were collected by filtration under vacuum. The crude product was purified by prep-HPLC using a C18 column (linear gradient 10-40% acetonitrile in 20 mM aqueous triethanolamine, pH 7.4). The prep-HPLC fractions were collected, and the organic solvent (acetonitrile) was evaporated under reduced pressure. The resulting aqueous solution was loaded onto a clean PDVB SPE column, washed with distilled water, then with a 0.1 M sodium acetate solution followed by distilled water. The product was eluted with

acetonitrile. The eluent was concentrated under reduced pressure, 385 mg was obtained as free base.

Synthesis of crystalline mono hydrochloride salt of (4S,4aS,5aR,12aS)-4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid amide (the “Crystalline Mono Hydrochloride Salt”).

[00106] Crude (4S,4aS,5aR,12aS)-4-dimethylamino-3, 10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l ,ll-dioxo-l,4,4a,5,5a,6,l l ,12a-octahydro-naphthacene-2-carboxylic acid amide (lOOg, app. 35% assay) was purified on preparative column chromatography. The desired fractions (8-10 liters) were combined and the pH was adjusted to 7.0-7.5 using ammonium hydroxide. This aqueous solution was extracted 3 times with dichloromethane (4 liters each time). The dichloromethane layers were combined and concentrated under reduced pressure. The residue was suspended in ethanol (800 ml) and 20 ml water was added. The pH was gradually adjusted to pH 1.6-1.3 using 1.25M hydrochloric acid in methanol and the mixture was stirred for 20-60 minutes at which point the free base was completely dissolved. The solution was concentrated under reduced pressure to 200-250 ml and was seeded with (4S,4aS,5aR,12aS)-4-dimethylamino-3,10, 12, 12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]- 1, 11-dioxo-l,4,4a,5,5a,6,l l,12a-octahydro-naphthacene-2-carboxylic acid amide mono HQ crystals (100-200 mg). The stirring was continued for 2-18 hours while the slurry was kept at <5°C. The resulting crystals were filtered, washed with ethanol (50 mL) and dried under reduced pressure to a constant weight. 20g crystalline (4S,4aS,5aR,12aS)-4-dimethylamino-3,10, 12, 12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]- 1, 11-dioxo-l,4,4a,5,5a,6,l l,12a-octahydro-naphthacene-2-carboxylic acid amide mono hydrochloride was isolated in > 90% purity and > 90% assay.

Synthesis of crystalline mono mesylate salt of (4S,4aS,5aR,12aS)-4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid (the “Crystalline Mesylate Salt”).

[00107] (4S,4aS,5aR,12aS)-4-dimethylamino-3, 10,12, 12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,l l,12a-octahydro-naphthacene-2-carboxylic acid amide free base (74mg) was suspended in ethanol (740μ1) and heated with stirring to 60°C (bath temperature). Methane sulfonic acid (1.1 eq, 167μ1 as 1M solution in THF) was added and most of the solid dissolved. After five minutes, the suspension was cooled to ambient temperature over approximately 1.75 hours (uncontrolled in oil bath). By 53 °C, solid had precipitated which was filtered at ambient temperature under reduced pressure. A further portion of ethanol (200μ1) was added to aid filtration, as the suspension was viscous. The cake was washed with n-hexane (400μ1) and air dried on filter for approximately 30 minutes to yield 59 mg (67% yield) of yellow solid.

Synthesis of crystalline mono sulfate salt of (4S,4aS,5aR,12aS)-4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid (the “Crystalline Sulfate Salt”).

[00108] (4S,4aS,5aR,12aS)-4-dimethylamino-3, 10,12, 12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,l l-dioxo-l,4,4a,5,5a,6,l l,12a-octahydro-naphthacene-2-carboxylic acid amide free base (86mg) was suspended in ethanol (500μ1) and heated with stirring to 63 °C (bath temperature) at which temperature most of the free base had dissolved. Sulfuric acid (1.1 eq, 194μ1 as 1M solution in water) was added and all of the solid dissolved. The solution was cooled to ambient temperature over approximately 1.75 hours (uncontrolled in oil bath) at which temperature no solid had precipitated. Methyl t-butyl ether (MtBE) was added as an antisolvent (4 x 50μ1). Each addition caused a cloud point, but the solid re-dissolved on stirring. The solution was stirred with a stopper for approximately 3 hours after which time solid precipitated. The solid was filtered under reduced pressure and washed with MtBE (3 x 200μ1) and air dried on filter for

approximately 45 minutes to yield 93 mg (90% yield) of yellow solid.

COMPARATIVE EXAMPLE 1

Synthesis of amorphous bis hydrochloride salt of (4S,4aS,5aR,12aS)-4-dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll-dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid amide.

[00109] (4S,4aS,5aR,12aS)-4-dimethylamino-3, 10,12, 12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,l l-dioxo-l,4,4a,5,5a,6,l l,12a-octahydro-naphthacene-2-carboxylic acid amide free base (1 g) was suspended in methanol (50 mL). The freebase was converted to the hydrochloride salt by adding an excess of methanolic HCl followed by under reduced pressure evaporation to give 1.1 g yellow solid: MS (Mz+1 = 488). 1H NMR (300 MHz, CD30D) δ 7.46 (d, 1H, J = 8.6 Hz), 6.81 (d, 1H, J = 8.6 Hz), 4.09 (d, 1H, J = 1.0 Hz), 3.79 (d, 1H, J = 13.1 Hz), 3.73 (d, 1H, J = 13.1 Hz), 3.36 (m, 1H), 3.27 (s, 3H), 3.08-2.95 (8H), 2.61 (s, 3H), 2.38 (t, 1H, J = 14.8), 2.22 (m, 1H), 1.64 (m, 1H). An XRPD pattern is shown in Figure 10 and a TGA and DSC curve overlaid are shown in Figure 11.

COMPARATIVE EXAMPLE 2

Synthesis of amorphous mono hydrochloride salt of (4S,4aS,5aR,12aS)-4- dimethylamino-3,10,12,12a-tetrahydroxy-7-[(methoxy(methyl)amino)-methyl]-l,ll- dioxo-l,4,4a,5,5a,6,ll,12a-octahydro-naphthacene-2-carboxylic acid amide.

[00110] A sample of Crystalline Mono Hydrochloride Salt (2.09 g) was dissolved in water (250 ml, 120 vols), filtered and frozen in a -78°C bath. Water was removed from the solidified sample using a lyophilizer for 110 hours to yield the amorphous mono hydrochloride salt as a fluffy yellow solid, that was confirmed to be amorphous by XRPD analysis .

PATENT

US 20130302442

PATENT

WO 2015153864

PATENT

WO 2018051102

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2003075857

References

External links

Sarecycline
Sarecycline.svg
Clinical data
Trade names Seysara
Identifiers
CAS Number
PubChem CID
ChemSpider
UNII
Chemical and physical data
Formula C24H29N3O8
Molar mass 487.51 g·mol−1
3D model (JSmol)

////////////Sarecycline, Seysara, WC-3035 FDA 2018, サレサイクリン , P-005672 , PTK-AR-01 , SC-1401, WC-3035,

AKN 028


img

AKN-028
CAS 1175017-90-9
Chemical Formula: C17H14N6
Molecular Weight: 302.33

N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine

N2-(1H-indol-5-yl)-6-(pyridin-4-yl)pyrazine-2,3-diamine

  • Originator Swedish Orphan Biovitrum
  • Developer Akinion Pharmaceuticals
  • Class Antineoplastics; Small molecules
  • Mechanism of Action Fms-like tyrosine kinase 3 inhibitors; Proto oncogene protein c-kit inhibitors
  • Phase I/II Acute myeloid leukaemia
  • 01 Mar 2016 Akinion Pharmaceuticals terminates phase I/II trial in Acute myeloid leukaemia in Czech Republic, Poland, Sweden and United Kingdom (NCT01573247)
  • 17 Sep 2015 AKN 028 is still in phase I/II trials for Acute myeloid leukaemia in Czech Republic, Poland and Sweden
  • 09 Apr 2014 AKN 028 is still in phase I/II trials for Acute myeloid leukaemia in Czech Republic, Poland and Sweden

AKN-028, a novel tyrosine kinase inhibitor (TKI), is a potent FMS-like receptor tyrosine kinase 3 (FLT3) inhibitor (IC(50)=6 nM), causing dose-dependent inhibition of FLT3 autophosphorylation. Inhibition of KIT autophosphorylation was shown in a human megakaryoblastic leukemia cell line overexpressing KIT. In a panel of 17 cell lines, AKN-028 showed cytotoxic activity in all five AML cell lines included. AKN-028 triggered apoptosis in MV4-11 by activation of caspase 3. In primary AML samples (n=15), AKN-028 induced a clear dose-dependent cytotoxic response (mean IC(50) 1 μM). However, no correlation between antileukemic activity and FLT3 mutation status, or to the quantitative expression of FLT3, was observed. Combination studies showed synergistic activity when cytarabine or daunorubicin was added simultaneously or 24 h before AKN-028. In mice, AKN-028 demonstrated high oral bioavailability and antileukemic effect in primary AML and MV4-11 cells, with no major toxicity observed in the experiment. (source: Blood Cancer J. 2012 Aug 3;2:e81. doi: 10.1038/bcj.2012.28.)

SYN

WO 2013/089636

Clip

Development of a Synthesis of Kinase Inhibitor AKN028

 R&D DepartmentMagle Chemoswed, P.O. Box 839, SE 201 80 Malmö, Sweden
 Recipharm OT ChemistryVirdings Allé 32 B, SE 754 50 Uppsala, Sweden
Org. Process Res. Dev., Article ASAP
DOI: 10.1021/acs.oprd.8b00092
*Telephone: +46 704473035. E-mail: johan.docera@gmail.com
Abstract Image

The novel tyrosine kinase inhibitor AKN028 has demonstrated promising results in preclinical trials. An expedient protocol for the synthesis of the compound at kilogram scale is described, including an SNAr reaction with high regioselectivity and a Suzuki coupling. Furthermore, an efficient method for purification and removal of residual palladium is described.

yellow or faint-orange powder. Mp 300 °C (dec.);

IR 3133 broad, 1689, 1597, 1554, 1480 cm–11H NMR (DMSO-d6) δ 11.01 (s, 1H), 8.62–8.50 (m, 2H), 8.22 (s, 1H), 8.15 (s, 1H), 8.06 (s, 1H), 7.89–7.82 (m, 2H), 7.39 (d, J = 2.0 Hz, 2H), 7.32 (t, J = 2.7 Hz, 1H), 6.77 (s, 2H), 6.42 (dd, J1 = 8.7 Hz, J2 = 2.0 Hz, 1H);

13C NMR (DMSO-d6) δ 149.9, 145.2, 145.0, 139.6, 132.8, 132.4, 132.2, 128.4, 127.6, 125.6, 118.7, 116.1, 111.2, 111.0, 101.0.

PATENT

 WO 2009095399

https://patentscope.wipo.int/search/ko/detail.jsf;jsessionid=074E97C06EF8C2088428DECCA2CD2EBA.wapp1nB?docId=WO2009095399&recNum=208&office=&queryString=&prevFilter=%26fq%3DOF%3AWO%26fq%3DICF_M%3A%22C07D%22%26fq%3DDP%3A2009&sortOption=Pub+Date+Desc&maxRec=3425

PATENT

WO 2013089636

https://patents.google.com/patent/WO2013089636A1/ko

Protein kinases are involved in the regulation of cellular metabolism, proliferation, differentiation and survival. The FLT-3 (fms-like tyrosine kinase) receptor is a member of the class III subfamily of receptor tyrosine kinases and has been shown to be involved in various disorders such as haematological disorders, proliferative disorders, autoimmune disorders and skin disorders.

In order to function effectively as an inhibitor, a kinase inhibitor needs to have a certain profile regarding its target specificity and mode of action. Depending on factors such as the disorder to be treated, mode of administration etc. the kinase inhibitor will have to be designed to exhibit suitable properties. For instance, compounds exhibiting a good plasma stability are desirable since this will provide a pharmacological effect of the compounds extending over time. Another example is oral administration of the inhibitor which may require that the inhibitor is transformed into a prodrug in order to improve the bioavailability.

WO 2009/095399 discloses pyrazine compounds acting as inhibitors of protein kinases, especially FTL3, useful in the treatment of haematological disorders, proliferative disorders, autoimmune disorders and skin disorders. This document discloses methods for manufacturing such compounds. However these methods are not suitable for large scale processes and the chemical yields are moderate. Furthermore, the compounds obtained by these methods are in amorphous form.

n one aspect of the invention, there is provided a process for preparing a compound of formula (I)

said process comprises the steps of:

a) reacting a compound of formula (1) with a compound of formula (2) in an inert solvent and in the presence of an (C1-6alkyl)3amine, providing a compound of formula (3):


, b) Suzuki coupling of a compound of formula (3) and a compound of formula (4) in an inert solvent and in the presence of a palladium catalyst and a base, providing a crude product comprising a compound of formula (I) and palladium

and

c) removing the palladium from the crude product in step b).

The compound of formula (I) may be obtained in amorphous or crystalline form using the processes outlined below.

Step 1:

Reaction of 2-amino-3,5-dibromopyrazine (1) and 5-aminoindole (2) in a

nucleophilic substitution reaction in the presence of a C1-6alkylamine and an inert polar solvent yields 3-bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine (3). Examples of inert polar solvents are DMSO, water and NEP. Examples of (C1-6alkyl)3amine are triethylamine, trimethylamine and tributylamine. The reaction may be performed at reflux temperature or at about 100-130°C.

Step 2:

A Suzuki coupling of 3-bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine) (3) and 4- pyridyl-boronic acid (4) in an inert polar solvent in the presence of a palladium catalyst and a base yields N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine (I) in amorphous form. Examples of inert solvents are DMF, water and DMA. Examples of palladium catalysts are Pd(dppf) and Pd(OAc)2-DTB-PPS. Example of a base is

K2CO3 The reaction may be performed under inert and oxygen-free atmosphere such as nitrogen or argon.

Heating may take place during step 1 and/or step 2. Steps 1 and 2 may be performed at reflux or in a temperature range of from 100 to 140°C, such as from 105 to 135°C, such as from 110 to 130°C, such as from 130-135°C, such as from 110-115ºC.

Step 3:

A compound of formula (I), also denominated N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine, in amorphous form may be dissolved in acetic acid (HOAc) after which potassium hydroxide (KOH) is added. The compound of formula (I) in amorphous form may be obtained from the process outlined in steps 1 and 2.

Alternatively, the compound of formula (I) may be obtained according to the process described in WO 2009/095399. The obtained crystalline form is removed from the slurry by, for instance, filtration. Step 3 may be repeated. Step 3 may be performed at a temperature of about 40°C followed by cooling to room temperature.

The process for preparing a compound according to formula (I) may comprise an additional step (step i) between step 2 and step 3 in order to remove palladium from the crude product of the compound of formula (I). The step comprises; forming a slurry comprising an acid and the compound according to formula (I) in a solvent, adding a siloxane compound to said slurry, removing the solvent from the slurry and adding an organic solvent, such as DMF and/or toluene, to the solid formed whereby a mixture is formed and then potassium hydroxide is added to the formed mixture, Alternatively, palladium may be removed from the crude product comprising (I) using a palladium scavenger such as TMT and/or 3-mercaptopropyl ethyl sulfide silica.

The crystalline form of the compound according to formula (I) may also be prepared from an amorphous form of the compound according to formula (I) by dissolving said amorphous form of the compound in a solvent mixture of

dichloromethane/methanol followed by evaporation of the solvent in a rotary evaporator. The amorphous form of the compound of formula (I) may obtained using the process disclosed in WO 2009/095399.

Example 1. Preparation of 5-Bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine (compound 3)

DMSO (10 L, 11 kg), 2-amino-3,5-dibromopyrazine (1) (4.5 kg, 17.8 mol, 1 eq.), 5- amino indole (2) (3.06 kg, 23.15 mol, 1.3 eq.) and triethylamine (7.4 L, 5.4 kg, 53.36 mol, 3 eq.) were charged to a reactor. The reaction mixture was heated to 95°C while agitated. After 12 hours, the heating was discontinued and the conversion was 88% of 2-amino-3,5-dibromopyrazine. The reaction was heated again to 95°C and

agitated for an additional 2.5 hours. There was no improvement in conversion. The reaction mixture was agitated at ambient temperature overnight. Triethylamine (3.5 kg) was removed under vacuum and the remaining reaction mixture was transferred to a stainless steel container from which it was charged into another reactor.

Subsequently, 18.4 kg of 50% acetic acid (aq.) was introduced over a period of 20 minutes under agitation, followed by purified water (61 L) charged over a period time of 60 minutes. The slurry was then filtered and the isolated material was washed with 2 x 20 L of 1% acetic acid (aq.).

The isolated 3-bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine) (3) was transferred to a drying cabinet and dried to invariable weight at 40 ±3°C, (19 hours), to afford 4.36 kg, 14.34 mol, 81 % yield, with a purity of 96% by HPLC.

The reaction temperature in the batch record was set to be 130-135°C. However, at 95°C the reaction mixture was at reflux.

Example 2. Preparation of N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3- diamine (compound I)

To a reactor was charged N,N-dimethylformamide (46.7 L, 45 kg), 4-pyridylboronic acid (4) (2.64 kg, 21.5 mol, 1.5 eq.) and 5-bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3- diamine (3) (4.36 kg, 14.3 mol). The reactor was then flushed with nitrogen prior to the charging of Pd(dppf)Cl2-catalyst (0.47 kg, 0.55 mol, 0.04 eq.). To reactor was then charged, over a period of 20 minutes, 24.9 kg of a 2 M solution of potassium carbonate (aq.). The reactor was flushed with nitrogen and heated under agitation to 110-115°C for 1.5 hours, after which 98.3% conversion of (3) was showed. The reaction mixture was quenched by addition of purified water (180 L) under vigorous agitation. The precipitated material was isolated on a hastalloy filter and washed with purified water (50 L), The isolated material was transferred to a drying cabinet and dried to invariable weight at 40 ±3°C (18 hours), to afford a compound of formula (5), i.e. a compound of formula (!) also denominated N-3-(1H-lndol-5-yl)-5-pyhdin-4-yl-pyrazine-2,3-diamine, (3.64 kg, 12.1 mol, 85 % yield).

During the process precipitated material was observed in the solutions, after the reactions, in both steps not previously seen in lab-scale. These impurities were not removed.

Example 3. Purification and crystallisation

In order to remove residual solvents from the material, two consecutive re-precipitations of the material from acetic acid were performed. This also gave crystallinity of the isolated substance. The purification is performed in order to remove palladium.

Purification

To a 1 L round bottomed flask was added 37.8 g of a compound according to formula (I) followed by 600 mL 2 M HOAc (aq.). The material was stirred at RT until a clear, dark red solution was obtained. To the solution was added 30 g Hyflo Super Celite and the slurry was filtered. The filter cake was washed with 25 mL 2 M HOAc

(aq) and 2×35 mL purified water. The obtained filtrate was transferred to a 2 L round bottomed flask containing 950 mL of Me-THF. The mixture was then stirred and heated to 40°C for 30 minutes. To the solution was then added 290 mL 8 M KOH (aq.) at 40°C and pH in the solution was 14.

The aqueous phase was removed and the organic phase washed with 2×100 mL of purified water. The remaining organic phase was then transferred to a 2 L round bottomed flask, followed by 95 mL of DMF, 20 g scavenger 3-Mercaptopropyl ethyl sulphide silica, Phosphonics LTD and 20 g scavenger 2-Mercaptoethyl ethyl sulfide silica purchased from Phosphonics LTD. The solution was vigorously stirred and heated at 60°C. A sample was withdrawn from the slurry after 12 hours, and showed 6 ppm of palladium remaining in the solution. The mixture was allowed to cool and was then filtered to remove the scavenger. The round bottomed flask and filter were rinsed with a mixture of 90 mL Me-THF and 10 mL DMF. Me-THF was then removed on a rotary evaporator and the remaining slurry was azeotropically dried with two portions of 100 mL toluene. To the remaining slurry was then added 85 mL of DMF to a total of 185 mL DMF (5ml DMF/g substance). To the clear solution was then added, slowly, while agitated, 1500 mL of toluene which produced a heavy precipitate. The slurry was filtered off and washed with 2×50 mL of toluene where after the material was dried overnight at 35°C under vacuum to afford 30.9 g of a compound according to formula (I) in a yield of 82%.

Crystallisation:

Example i

1. First re-precipitation

The N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine material (30.9 g) was added to a 1 L round bottomed flask and 450 mL 2 M HOAc (aq.) was added. The slurry was agitated and heated to 40°C for 1 hour, until the material had dissolved. To the solution was then added 158 mL 8 M KOH (aq.) at 40°C. The pH in the solution was 11.4. The slurry was then allowed to cool to 25°C and filtered. The filter cake was washed with 3x 80 mL of purified water and the material was dried overnight at 95°C under vacuum to afford 28.7g N-3-(1H-indol-5-yl)-5-pyridin-4-yl- pyrazine-2,3-diamine in a yield of 93%.

2. Second re-precipitation

N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine material (28.7 g) was added to a 1L round bottomed flask and 430 mL 2 M HOAc (aq) was added. The slurry was agitated and heated to 40°C for 1 hour, until the material had dissolved. To the solution was then added 15 mL 8M KOH (aq) at 40°C. The pH in the solution was 12.3. The slurry was then allowed to cool to 25°C and filtered. The filter cake was washed with 5×50 mL of purified water, and the solid was then dried overnight at 95°C under vacuum to afford 28.3 g N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3- diamine in a yield of 99%.

Example ii

The N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine material (2.1 kg, 7 mol) was added to a reactor, followed by 2M HOAc (aq.) (59.6 L, 60.2 kg) . The solution in the reactor was then heated to 40°C and stirred for 20 minutes. To the clear solution was then charged, slowly, 30% KOH (aq.) (25 kg) under vigorous agitation. The slurry was agitated for 15 minutes. pH in the solution was 6.2, and a total of 1.5 kg 30% KOH (aq.) was then added to the solution to give pH 12.1. The precipitated material was isolated on a Hastelloy filter and washed with purified water (5×30 L). The solid was then transferred to a drying cabinet and dried to invariable weight at 85 ±3°C under vacuum (16 hours; a sample was withdrawn after 16 hours, showing 1400 ppm HOAc and 75 ppm DMF), to afford N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine (2.0 kg, 7 mol, 95 % yield).

Hence, N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine is obtained in an uniform crystalline form, which was achieved by precipitating the product from aqueous acetic acid by introduction of aqueous potassium hydroxide.

Example 5. Synthesis of 5-Bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine (compound 3)

2-Amino-3,5-dibromopyrazine (45 g, 1.0 eq.), 5-aminoindole (30,6 g, 1.3 eq.), 67.5 mL NEP, i.e. 1-ethyl-2-pyrrolidone, and 74.5 mL triethylamine were added to a 250 mL reactor. The jacket temperature was set to 130°C and the reaction mixture was stirred for 22 h. HPLC after 22 h showed 87% conversion of the 2-amino-3,5-dibromopyrazine. After 24 h HPLC showed 92% conversion and the reaction slurry was cooled to 80°C and quenched by addition of addition of 50% HOAc(aq) and water. The obtained slurry was then allowed to cool to room temperature over night while agitated. The material was isolated on a glass filter funnel and was washed with water. The material was dried at 80 °C under vacuum until dry to afford 71% of the compound 5-bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine as a dark brown powder. The purity was 99.8% as measured by HPLC.

Example 6. Synthesis of N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine (Compound I)

5-Bromo-N-3-(1H-indol-5-yl)-pyrazine-2,3-diamine (15.0 g, 49 mmol, 1.0 eq.), 4-pyridyl boronic acid (6.6 g, 59 mmol, 1.2 eq.), Pd(OAc)2 (166 mg, 0.74 mmol, 0.015 eq.), DTB-PPS, i.e. 3-(di-tert-butylphosphino)propane-1-sulfonic acid, (199 mg, 0.74 mmol, 0.015 eq.), and DMA, i.e. N,N-dimethylacetamide, (75 mL) were added to a three-necked round-bottomed flask equipped with a mechanical stirrer,

thermometer, and a nitrogen atmosphere. Through a septa was added 2M K2CO3 (aq) (27 ml, 54 mmol, 1.1 eq.) with a syringe. The temperature was increased to 100 °C. Samples for HPLC-analysis of the conversion were drawn and when the conversion had reached 100% the temperature was cooled to 25 °C. At that temperature a water solution of 0.5 M L-cysteine (150 ml) was added by a syringe pump over 1 hour with a rate of 2.5 mL/minute. After 3 hours maturing time at room temperature the material was isolated on a glass filter funnel and was washed with water. The material was dried at 40 °C under vacuum over the weekend, and 15 grams of N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine (101%) were obtained as a brown powder.

Example 7. Purification of N-3-(1H-indol-5-yl)-5-pyridin-4-yl-pyrazine-2,3-diamine (Compound I)

The crude (7.0 g, 23 mmol) and 2M HOAc (98 mL) was added to a 250 mL round-bottomed flask. To this was added TMT, i.e. trithiocyanuric acid, (1.4 g) and SPM32, i.e. 3-mercaptopropyl ethyl sulfide silica, (1.4 g). The mixture was stirred in room temperature for 24 hours. After 24 hour a polish filtration through hyflo super cel was performed. To the clear filtrate was added 50 mL 5 M KOH(aq) under 15 minutes to precipitate the product. After 18 hours maturing time at room temperature the material was isolated on a glass filter funnel and was washed with 2×20 mL water. The first was being a slurry wash and the second a displacement wash. The material was dried at 40 °C under vacuum over the weekend, and 3.9 grams (56%) was obtained as a light yellow powder. The Pd content was 3.7 ppm.

PATENT

US 8436171

PATENT

WO 2016008433

PATENT

WO 2016015604

PATENT

WO 2016015597

PATENT

WO 2016015605

PATENT

WO 2016015598

PATENT

WO 2017146794

PATENT

WO 2017146795

https://patents.google.com/patent/WO2017146795A1/en

PATENT

US 20180071302

REFERENCES

1: Eriksson A, Hermanson M, Wickström M, Lindhagen E, Ekholm C, Jenmalm Jensen A, Löthgren A, Lehmann F, Larsson R, Parrow V, Höglund M. The novel tyrosine kinase  inhibitor AKN-028 has significant antileukemic activity in cell lines and primary cultures of acute myeloid leukemia. Blood Cancer J. 2012 Aug 3;2:e81. doi: 10.1038/bcj.2012.28. PubMed PMID: 22864397; PubMed Central PMCID: PMC3432483.

////////////AKN028 , AKN-028 , AKN 028, phase 2, Swedish Orphan Biovitrum,  Akinion Pharmaceuticals,  Acute myeloid leukaemia

NC1=NC=C(C2=CC=NC=C2)N=C1NC3=CC4=C(NC=C4)C=C3

FDA approves a new antibacterial drug to treat a serious lung disease using a novel pathway to spur innovation


FDA approves a new antibacterial drug to treat a serious lung disease using a novel pathway to spur innovation

First drug granted approval under FDA’s Limited Population Pathway for Antibacterial and Antifungal Drugs, instituted to spur development of antibiotics for unmet medical needs

The U.S. Food and Drug Administration today approved a new drug, Arikayce (amikacin liposome inhalation suspension), for the treatment of lung disease caused by a group of bacteria, Mycobacterium avium complex (MAC) in a limited population of patients with the disease who do not respond to conventional treatment (refractory disease).

MAC is a type of nontuberculous mycobacteria (NTM) commonly found in water and soil. Symptoms of disease in patients with MAC include persistent cough, fatigue, weight loss, night sweats, and occasionally shortness of breath and coughing up of blood.

September 28, 2018

Release

The U.S. Food and Drug Administration today approved a new drug, Arikayce (amikacin liposome inhalation suspension), for the treatment of lung disease caused by a group of bacteria, Mycobacterium avium complex (MAC) in a limited population of patients with the disease who do not respond to conventional treatment (refractory disease).

MAC is a type of nontuberculous mycobacteria (NTM) commonly found in water and soil. Symptoms of disease in patients with MAC include persistent cough, fatigue, weight loss, night sweats, and occasionally shortness of breath and coughing up of blood.

“As bacteria continue to grow impervious to currently available antibiotics, we need to encourage the development of drugs that can treat resistant infections. That means utilizing novel tools intended to streamline development and encourage investment into these important endeavors,” said FDA Commissioner Scott Gottlieb, M.D. “This approval is the first time a drug is being approved under the Limited Population Pathway for Antibacterial and Antifungal Drugs, and it marks an important policy milestone. This pathway, advanced by Congress, aims to spur development of drugs targeting infections that lack effective therapies. We’re seeing a lot of early interest among sponsors in using this new pathway, and it’s our hope that it’ll spur more development and approval of antibacterial drugs for treating serious or life-threatening infections in limited populations of patients with unmet medical needs.”

Arikayce is the first drug to be approved under the Limited Population Pathway for Antibacterial and Antifungal Drugs, or LPAD pathway, established by Congress under the 21st Century Cures Act to advance development and approval of antibacterial and antifungal drugs to treat serious or life-threatening infections in a limited population of patients with unmet need. Approval under the LPAD pathway may be supported by a streamlined clinical development program. These programs may involve smaller, shorter or fewer clinical trials. As required for drugs approved under the LPAD pathway, labeling for Arikayce includes certain statements to convey that the drug has been shown to be safe and effective only for use in a limited population.

Arikayce also was approved under the Accelerated Approval pathway. Under this approach, the FDA may approve drugs for serious or life-threatening diseases or conditions where the drug is shown to have an effect on a surrogate endpoint that is reasonably likely to predict a clinical benefit to patients. The approval of Arikayce was based on achieving three consecutive negative monthly sputum cultures by month six of treatment. The sponsor of Arikayce will be required by the FDA to conduct an additional, post-market study to describe the clinical benefits of Arikayce.

The safety and efficacy of Arikayce, an inhaled treatment taken through a nebulizer, was demonstrated in a randomized, controlled clinical trial where patients were assigned to one of two treatment groups. One group of patients received Arikayce plus a background multi-drug antibacterial regimen, while the other treatment group received a background multi-drug antibacterial regimen alone. By the sixth month of treatment, 29 percent of patients treated with Arikayce had no growth of mycobacteria in their sputum cultures for three consecutive months compared to 9 percent of patients who were not treated with Arikayce.

The Arikayce prescribing information includes a Boxed Warning regarding the increased risk of respiratory conditions including hypersensitivity pneumonitis (inflamed lungs), bronchospasm (tightening of the airway), exacerbation of underlying lung disease and hemoptysis (spitting up blood) that have led to hospitalizations in some cases. Other common side effects in patients taking Arikayce were dysphonia (difficulty speaking), cough, ototoxicity (damaged hearing), upper airway irritation, musculoskeletal pain, fatigue, diarrhea and nausea.

The FDA granted this application Fast Track, Breakthrough Therapy, Priority Review, and Qualified Infectious Disease Product (QIDP) designations. QIDP designation is given to antibacterial products that treat serious or life-threatening infections under the Generating Antibiotic Incentives Now (GAIN) title of the FDA Safety and Innovation Act. Arikayce also received Orphan Drug designation, which provides additional incentives to assist and encourage the development of drugs for rare diseases.

The FDA granted approval of Arikayce to Insmed, Inc. of Bridgewater, NJ.

/////////////////// Arikayce, amikacin liposome inhalation suspension, fda 2018, Fast Track, Breakthrough Therapy, Priority Review, and Qualified Infectious Disease Product, QIDP, Generating Antibiotic Incentives Now, GAIN,

Efonidipine, エホニジピン


Efonidipine structure.svg

ChemSpider 2D Image | Efonidipine | C34H38N3O7P

Efonidipine.png

Efonidipine

  • Molecular FormulaC34H38N3O7P
  • Average mass631.655 Da
  • エホニジピン
  • CAS 111011-63-3; FREE FORM
(±)-Efonidipine
Image result for Efonidipine
Molecular Formula: C36H45ClN3O8P
Molecular Weight: 714.193 g/mol

LD50:> 5 g/kg (R, p.o.)

  • Synonyms:NZ-105
  • ATC:C08CA
Efonidipine hydrochloride monoethanolate  111011-76-8 [RN],エホニジピン塩酸塩エタノール付加物
CAS 111011-63-3; FREE FORM
2-(N-Benzylanilino)ethyl (±)-1,4-dihydro-2,6-dimethyl-4-(m-nitrophenyl)-5-phosphononicotinate Cyclic 2,2-Dimethyltrimethylene Ester
2-[Benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxido-1,3,2-dioxaphosphinan-2-yl)-2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydro-3-pyridinecarboxylate
2-[Benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxido-1,3,2-dioxaphosphinan-2-yl)-2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3-carboxylate
3-Pyridinecarboxylic acid, 5-(5,5-dimethyl-2-oxido-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-, 2-[phenyl(phenylmethyl)amino]ethyl ester
40ZTP2T37Q
5-(5,5-Dimethyl-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylic Acid 2-[Phenyl(phenylmethyl)amino]ethyl Ester P-Oxide
Landel [Trade name]
UNII:40ZTP2T37Q
2-(N-benzylanilino)ethyl 5-(5,5-dimethyl-2-oxo-1,3,2$l^{5}-dioxaphosphinan-2-yl)-2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3-carboxylate

Efonidipine Hydrochloride Ethanolate Bulk & Tablets 10 mg/20mg/40mg, 

Indicated for the management of
• Hypertension
• Renal parenchymal hypertension
• Angina
CDSCO approved INDIA 28.08.2017 
Launched – 1994, Shionogi Zeria
Efonidipine (INN) is a dihydropyridine calcium channel blocker marketed by Shionogi & Co. of Japan. It was launched in 1995, under the brand name Landel. The drug blocks both T-type and L-type calcium channels [A7844, A32001]. It has also been studied in atherosclerosis and acute renal failure [A32001]. This drug is also known as NZ-105, and several studies have been done on its pharmacokinetics in animals [L1456].

Efonidipine (INN) is a dihydropyridine calcium channel blocker marketed by Shionogi & Co. of Japan. It was launched in 1995, under the brand name Landel (ランデル). The drug blocks both T-type and L-type calcium channels.[1] Drug Controller General of India (DCGI) granted approval to M/s. Zuventus pharma Ltd for marketing efonidipine under brand name Efnocar in India .[2]

Structure Activity Relationship

Efonidipine is a dual Calcium Channel Blocker (L & T-type). It has a unique chemical structure. The phosphonate moiety (Figure 1) at the C5 position of the dihydropyridine ring is considered to be important for the characteristic pharmacological profile of the drug. (figure-1)

Figure-1:Efonidipine: Chemical Structure

Mechanism of action

Efonidipine, a new generation dihydropyridine (DHP) calcium channel blocker, inhibits both L-type and T-type calcium channels.[1]

Pharmacodynamics

  • Efonidipine exhibits antihypertensive effect through vasodilatation by blocking L-type and T-type calcium channels.[1]
  • Efonidipine has a negative chronotropic effect. Working on sino atrial node cells by inhibiting T-type calcium channel activation, Efonidipine prolongs the late phase-4 depolarization of the sino atrial node action potential and suppresses an elevated HR. The negative chronotropic effect of Efonidipine decreases heart rate, myocardial oxygen demand and increases coronary blood flow.[3]
  • Efonidipine increases coronary blood flow by blocking L & T-type calcium channels and attenuates myocardial ischaemia.[4]
  • By reducing synthesis and secretion of aldosterone, Efonidipine prevents hypertrophy and remodeling of cardiac myocytes.[5]
  • Efonidipine increases glomerular filtration rate without increasing intra-glomerular pressure and filtration fraction. This prevents hypertension induced renal damage.[6]
  • Efonidipine prevents Rho-kinase and NFB induced renal parenchymal fibrosis and provides long term renal protection.[7][8]
  • Efonidipine suppresses renin secretion from the juxta glomerular apparatus in the kidneys.[9]
  • Efonidipine enhances sodium excretion from the kidneys by suppressing aldosterone synthesis and secretion from the adrenal glands. Aldosterone induced renal parenchymal fibrosis is suppressed by Efonidipine.[5]
  • Efonidipine prevents NFB induced hypertrophy and inflammation in the renal vasculature and protects the kidneys.[7]
  • Efonidipine protects against endothelial dysfunction due to its anti-oxidant activity and by restoring NO bioavailability.[10][11]
  • Efonidipine has anti-atherogenic activity and protects the blood vessels from atherosclerosis.[12]
  • Efonidipine lowers blood pressure in cerebral resistance vessels and prevents hypertension induced brain damage.[4]

Pharmacokinetics

Absorption

Peak plasma concentration is achieved in about 1.5 to 3.67 hours after administration. Half life is approximately 4 hours. The pharmacokinetic parameters of Efonidipine are depicted in Table-1.

Table 1: PK Parameters in Adult Healthy Male Subjects

Variable Efonidipine
Mean Range
Cmax(ng/ml) 36.25 9.66-66.91
Tmax (hour) 2.59 1.50-3.67
T1/2 (hour) 4.18 2.15-6.85

*Data on file

Long Duration of Action

Efonidipine has a slow onset and a long duration of action. This unique characteristic of Efonidipine is because of the following reasons:[13]

  1. High lipophilicity of Efonidipine allows it to enter the phospholipid rich cell membrane and access the dihydropyridine binding site of the Ca2+ channels.
  2. Tight binding to the dihydropyridine receptors.
  3. The dissociation constant of Efonidipine from dihydropyridine receptors is very low (0.0042/min/nM), signifying very slow dissociation from the receptors. This explains the long duration of action of Efonidipine.

Metabolism

Efonidipine is primarily metabolized in the liver. The important metabolites are N-dephenylated Efonidipine (DPH), deaminated Efonidipine (AL) and N-debenzylated Efonidipine (DBZ). DBZ and DPH exhibit activity as calcium antagonists. The vasodilating properties of DBZ and DPH were about two-thirds and one-third respectively than that of the parent compound. Results suggest that the majority of the pharmacological effect after oral dosing of Efonidipine hydrochloride in man is due to unchanged compound and its metabolites make a small contribution to the pharmacological effect.[14]

Elimination

Biliary route is the main pathway of excretion. No significant amount of unchanged drug was excreted in urine. In the urine collected for 24 h after an oral dosing, 1.1 % of the dose was excreted as deaminated Efonidipine, and 0.5% as a pyridine analogue of deaminated Efonidipine.

Indications

  • Essential hypertension and renal parenchymal hypertension
  • Angina

Dosage and Administration

  • Essential hypertension and renal parenchymal hypertension: 20-40 mg orally once daily. A dose of up to 80mg/day is seen to be safe and effective in clinical trials.[15][16]
  • Angina: 40 mg/day.

Contraindications

  • Contraindicated in patients hypersensitive to Efonidipine or any of the excipients
  •  It is also contraindicated in pregnancy and lactation.

Precautions

  •  Should be administered with caution in patients with hepatic impairment
  • Dose adjustment may be required in elderly as hypotension can occur
  •  Efonidipine may worsen clinical condition in patients with sinus bradycardia, sinus arrest or sinus node dysfunction
  • As dizziness can occur due to hypotensive action, one should be careful while operating machines, with aerial work platforms and driving of a motor vehicle
  • Drug should not be stopped abruptly. Discontinuation should be gradual and under supervision of a qualified physician

Drug Interactions

  • Other anti-hypertensive agents: Efonidipine enhances the antihypertensive action additively and may produce hypotension and shock. Blood pressure should be monitored regularly to adjust dose of concomitant drugs.
  •  Cimetidine: Cimetidine inhibits CYP450 enzymes involved in metabolism of CCBs. Blood concentration of calcium channel antagonists increase leading to higher incidence of side effects (hot flushes).
  • Grape fruit juice: Grapefruit juice suppresses enzymes metabolizing calcium channel antagonists (cytochrome P450) and reduces the clearance. Thus, there is a possibility that blood concentration of the drug may increase and the anti-hypertensive effect is enhanced.
  • Tacrolimus: Efonidipine inhibits metabolic enzymes involved in Tacrolimus metabolism and reduces its clearance. So, increase in blood concentration of Tacrolimus can occur.

Adverse Drug Reactions

The common side effects are hot flushes, facial flushing and headache. In addition, elevation in serum total cholesterol, ALT (SGPT), AST (SGOT) and BUN may occur. Frequent urination, pedal edema, increased triglycerides occurs in less than 0.1%.[17]

Lesser incidence of pedal edema (< 0.1%)

One common adverse effect of the L-type Ca2+ channel blockers like Amlodipine is vasodilatory Pedal edema. Combined L-/T-type Ca2+ channel blockers, such as Efonidipine, display antihypertensive efficacy similar to their predecessors (Amlodipine) with much less propensity of pedal edema formation. Efonidipine equalizes the hydrostatic pressure across the capillary bed through equal arteriolar and venular dilatation, thus reducing vasodilatory edema. These incremental microcirculatory benefits of efonidipine over the conventional L-type Ca2+ channel blockers (Amlodipine) are likely attributed to their additional T-type Ca2+ channel blocking properties and the increased presence of T-type Ca2+channels in the microvasculature (e.g. arterioles, capillaries, venules etc).[18]

Among the CCBs, Efonidipine (<0.1%)[17] has lowest incidence of pedal edema compared to amlodipine ( 5-16%)[19], cilnidipine (5%)[20], benidipine (5%)[21] and azelnidipine (15.5%).[22]

Use in Special Population

Administration to Elderly

The drug should be started at low dose (20 mg/day) in elderly. Patient should be carefully observed for development of hypo-tension. Dose may be halved if there is intolerance to the 20 mg/day dosage regimen.

Pregnancy and Lactation

The drug should not be administered to pregnant women and women suspected of being pregnant. Administration to lactating women should be avoided unless benefit significantly surpasses the risk to the child. Mothers on Efonidipine treatment should avoid breast feeding.

Pediatric Use

Safety of Efonidipine in low birth weight infants, newborns, infants and children has not been established.

Efonidipine-The Best in Class

Efonidipine is unique among clinically available CCBs. Its antihypertensive efficacy is superior or at par with other CCBs. But, in terms of pleiotropic effects leading to enhanced cerebral, cardiac and renal protection, Efonidipine scores over the other CCBs.

Advantages over Amlodipine

1.      Better renoprotection by:

  • Dual channel blockade [1]
  • Prevention of Rho-kinase and NFkB induced tubulointerstitial fibrosis[23][24]
  • Reduction of synthesis and secretion of aldosterone from the adrenal cortex[25]

2.       Preferred in  angina  with hypertension due to negative chronotropic action[26]

3.       Better control of reflex tachycardia[3]

4.       Reduces cardiac remodelling, arterial stiffness and prevents atherogenesis[27]

5.       More useful in patients with diabetes & nephropathy[28]

6.       Better protection  against cardiac hypertrophy by significant reduction in LVMI[29]

7.       Less adverse effects compared to Amlodipine[30]

8.       Reduces endothelial dysfunction and oxidative stress(anti-oxidant property)[10]

Advantages over Cilnidipine

1.       Strong negative chronotropic effect (less tachycardia) compared to Cilnidipine[3]

2.       Significant improvement in exercise tolerance.[31]Better choice in hypertensive patients with angina.

3.       Better BP control by marked urinary Na+ excretion[32]

4.       Better renoprotection by:

  • a.      Suppression of plasma renin release[33]
  • b.     Prevention of Rho-kinase and NFkB induced tubulointerstitial fibrosis[34][35]
  • c.      Reduction of synthesis and secretion of aldosterone from the adrenal cortex[5]

5.       Better choice in diabetic hypertensives[36]

6.       Prevents cardiac remodelling by suppression of aldosterone secretion[5]

7.       Superior anti-oxidant activity[10]

8.       Less adverse effects compared to Cilnidipine[30]

Advantages over Benidipine

L & T-type CCBs have invoked a lot of interest in the management of hypertension because of their unique pharmacological profile. Several novel agents have been developed including Azelnidipine, Barnidipine, Benidipine, Efonidipine, Manidipine and Nilvadipine. Among all the agents, Efonidipine has emerged as the best among its peers. The advantages of Efonidipine over Benidipine are summarized below.

1. More selective blockade of T-type calcium channels [37][38]

2. More balanced renal arteriolar dilatation than benidipine[37][38]

3. Superior anti-proteinuric effect [15]

4. Greater reduction of serum aldosterone [39]

5. Renoprotection by reducing plasma renin unlike Benidipine [39]

6. Greater negative chronotropic effect

7. Efonidipine has anti-platelet activity[12]

8. Efonidipine reduces Insulin Resistance [40]

9. Significantly lower incidence of pedal edema & constipation compared to Benidipine

A new synthesis of efonidipine has been described: The cyclization of 2,2-dimethylbutane-1,4-diol (I) with triethyl phosphite (II) by heating at 100 C gives 2-methoxy-5,5-dimethyl-1,3,2-dioxaphosphorinan (III), which, by treatment with iodoacetone (IV) in refluxing ether, yields 2-acetonyl-5,5-dimethyl-1,3,2-dioxaphosphorinan-2-one (V). The condensation of (V) with 3-nitrobenzaldehyde (VI) by means of piperidine in acetic acid affords 3-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-4-(3-nitrophenyl)-3-buten-2-one (VII), which is finally cyclized with 3-amino-2-propenoic acid 2-(N-benzyl-N-phenylamino)ethyl ester (VIII) in refluxing toluene.ReferencesChem Pharm Bull 1992,40(9),2362

A new synthesis for (4S)-efonidipine has been described: The reaction of 5,5-dimethyl-2-(2-oxopropyl)-1,3,2-dioxaphosphorinan-2-one (I) with dimorpholino(3-nitrophenyl)methane (II) by means of trifluoroacetic acid in hot toluene gives 5,5-dimethyl-2-[1-acetyl-2-(3-nitrophenyl)vinyl]-1,3,2-dioxaphosphorina n-2-one (III), which is cyclized with 3-aminocrotonic acid 2(S)-methoxy-2-phenylethyl ester (IV) in refluxing toluene; the recrystallization of the resulting product affords 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-2,6-dimethyl-4(S)-(3-nitrophenyl)-1,4-dihydropyridine-3-carboxylic acid 2(S)-methoxy-2-phenylethyl ester (V). The protection of the NH group of (V) with chloromethyl methyl ether and NaH in THF yields the N-methoxymethyl derivative (VI), which is transesterified with 2-(N-benzyl-N-methylamino)ethanol (VII) and NaH in DMSO, giving the protected final product (VIII). Finally, this compound is deprotected with HCl in ethanol.

An enantioselective synthesis of efonidipine has been described: The enantioselective hydrolysis of 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-2,6-dimethyl-4-(3-n itrophenyl)-1,4-dihydropyridine-3-carboxylic acid propionyloxymethyl ester (I) with lipase AH in 2,5-dimethyltetrahydrofuran saturated with water gives the corresponding free acid of the (S)-isomer (III), while the propionyloxymethyl ester of the (R)-isomer (II) remains undisturbed. After chromatographic separation, the (R)-ester (II) is hydrolyzed with NaOH in methanol to the (R)-acid (IV). Finally, both enantiomerically pure acids (III) and (IV) are separately esterified with 2-(N-benzyl-N-phenylamino)ethanol in the usual way

CLIP

PAPER

Synthesis of 1,4-dihydropyridine-5-phosphonates and their calcium antagonistic and antihypertensive activities: Novel calcium-antagonist 2-[benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate hydrochloride ethanol (NZ-105) and its crystal structure
Chem Pharm Bull 1992, 40(9): 2362

PATENT

IN 201501586

http://ipindiaservices.gov.in/PatentSearch/PatentSearch/ViewPDF

  1. Jump up to:a b c d Tanaka H, Shigenobu K (2002). “Efonidipine hydrochloride: a dual blocker of L- and T-type Ca2+ channels”. Cardiovasc. Drug Rev20 (1): 81–92. PMID 12070536.
  2. Jump up^http://www.cdsco.nic.in/writereaddata/Minutes%20of%2034th%20SEC%20Cardiovascular%20&%20Renal%2008_11_2016.pdf
  3. Jump up to:a b c Masumiya H, Shijuku T, Tanaka H, Shigenobu K. Inhibition of myo¬cardial L- and T-type Ca2+ currents by efonidipine: possible mecha¬nism for its chronotropic effect. Eur J Pharmacol. 1998; 349: 351-7.
  4. Jump up to:a b Masuda Y, Tanaka S. Efonidipine Hydrochloride: A New Calcium Antagonist. Cardiovascular Drug Reviews. 1994; 12 ( 2): 123-135.
  5. Jump up to:a b c d Ikeda K, Isaka T, Fujioka K, Manome Y, Tojo K. Suppression of Aldosterone Synthesis and Secretion by Ca2+ Channel Antagonists. International Journal of Endocrinology. 2012.
  6. Jump up^ Hayashi K, et al. T-Type Ca Channel Blockade as a Determinant of Kidney Protection. Keio J Med. 2010; 59(3): 84-95
  7. Jump up to:a b Hayashi M, Yamaji Y, Nakazato Y, Saruta T. The effects of calcium channel blockers on nuclear factor kappa B activation in the mesangial cells. Hypertens Res. 2000;23:521–525.
  8. Jump up^ Sugano N, Sugano N, Wakino S, Tatematsu S, Homma K, Yoshioka K, Hasegawa K, Utsunomiya Y, Tokudome G, Hosoya T, Saruta T, Hayashi K. Role of T-type Ca2 channels (TCCs) as a determinant of Rho-kinase activation and epithelial-mesenchymal transition (EMT) in renal injury. J Hypertens. 2006;24(suppl 6):128
  9. Jump up^ Baylis C, Qiu C, Engels K. Comparison of L-type and mixed L- and T-type calcium channel blockers on kidney injury caused by deoxycorticosterone-salt hypertension in rats. Am J Kidney Dis. 2001;38: 1292–1297.
  10. Jump up to:a b c Sasaki H, Saiki A, Endo K, Ban N, Yamaguchi T, Kawana H, Nagayama D, Ohhira M, Oyama T, Miyashita Y, Shirai K. Protective effects of efonidipine, a T- and L-type calcium channel blocker, on renal function and arterial stiffness in type 2 diabetic patients with hypertension and nephropathy. J Atheroscler Thromb. 2009 Oct; 16(5): 568-75.
  11. Jump up^ Oshima T, Ozono R, Yano Y, Higashi Y, Teragawa H, Miho N, Ishida T, Ishida M, Yoshizumi M, Kambe M. Beneficial effect of T-type calcium channel blockers on endothelial function in patients with essential hypertension. Hypertens Res. 2005 Nov;28(11):889-94.
  12. Jump up to:a b Nomura S, Kanazawa S, Fukuhara S. Effects of efonidipine on platelet and monocyte activation markers in hypertensive patients with and without type 2 diabetes mellitus. J Hum Hypertens. 2002 Aug;16(8):539-47.
  13. Jump up^ Yamashita T, Masuda Y, et al. NZ-105, a New 1,4-Dihydropyridine Derivative: Correlation between Dihydropyridine Receptor Binding and Inhibition of Calcium Uptake in Rabbit Aorta. Japan J Pharmacol. 1991; 57: 337-348.
  14. Jump up^ Nakabeppu H, et al.Metabolism of Efonidipine in Man.Xenobiotica.1995 Aug;229-239.
  15. Jump up to:a b Hayashi K, Kumagai H, Saruta T. Effects of efonidipine and ACE inhibitors on proteinuria in human hypertension with renal impairment. Am J Hypertens. 2003;16:116 –122
  16. Jump up^  Oh IY, Seo MK, Lee HY, Kim SG, Kim KS, Kim WH, Hyon MS, Han KR, Lim SJ, Kim CH. Beneficial Effect of Efonidipine, an L- and T-Type Dual Calcium Channel Blocker, on Heart Rate and Blood Pressure in Patients With Mild-to-Moderate Essential Hypertension. Korean Circ J. 2010 Oct;40(10):514-9.
  17. Jump up to:a b http://www.kegg.jp/medicus-bin/japic_med?japic_code=00044638. Missing or empty |title= (help)
  18. Jump up^ Ge W, Ren J. Combined L-/T-type calcium channel blockers: ready for prime time. Hypertension. 2009 Apr;53(4):592-4. doi: 10.1161/HYPERTENSIONAHA.108.127548. Epub 2009 Feb 23. PubMed PMID 19237678.
  19. Jump up^ Osterloh IH. An update on the safety of amlodipine. J Cardiovasc Pharmacol.1991;17 Suppl 1:S65-8.
  20. Jump up^ http://www.kegg.jp/medicus-bin/japic_med?japic_code=00062065. Missing or empty |title= (help)
  21. Jump up^ http://www.kegg.jp/medicus-bin/japic_med?japic_code=00005939. Missing or empty |title= (help)
  22. Jump up^ Takihata M, Nakamura A, Kondo Y, Kawasaki S, Kimura M, Terauchi Y. Comparison of Azelnidipine and Trichlormethiazide in Japanese Type 2 Diabetic Patients with Hypertension: The COAT Randomized Controlled Trial. PLoS One. 2015 May 4;10(5):e0125519.
  23. Jump up^ Song I, KimD, Choi S, Sun M, Kim Y, Shin HS. Role of the α1g T-type calcium channel in spontaneous absence seizures in mutant mice. J Neurosci. 2004; 24: 5249–5257.
  24. Jump up^ Lory P, Bidaud I, Chemin J. T-Type calcium channels in differentiation and proliferation. Cell Calcium. 2006; 40: 135–146.
  25. Jump up^ Ikeda K, Isaka T, Fujioka K, Manome Y, Tojo K. Suppression of Aldosterone Synthesis and Secretion by Ca2+ Channel Antagonists. International Journal of Endocrinology. 2012.
  26. Jump up^ Oh IY, Seo MK, Lee HY, Kim SG, Kim KS, Kim WH, Hyon MS, Han KR, Lim SJ, Kim CH. Beneficial Effect of Efonidipine, an L- and T-Type Dual Calcium Channel Blocker, on Heart Rate and Blood Pressure in Patients With Mild-to-Moderate Essential Hypertension. Korean Circ J. 2010 Oct;40(10):514-9.
  27. Jump up^ Catena C, Colussi G, Marzano L, Sechi LA. Aldosterone and the heart: from basic research to clinical evidence. Horm Metab Res. 2012;44:181– 187. 
  28. Jump up^  Sasaki H, Saiki A, Endo K, Ban N, Yamaguchi T, Kawana H, Nagayama D, Ohhira M, Oyama T, Miyashita Y, Shirai K. Protective effects of efonidipine, a T- and L-type calcium channel blocker, on renal function and arterial stiffness in type 2 diabetic patients with hypertension and nephropathy. J Atheroscler Thromb. 2009 Oct; 16(5): 568-75.
  29. Jump up^  Saito T, Fujii K, Takizawa T, Toyosaki T, Kuwabara Y, Kobayashi S, Ichikawa H, Karaki A, Yamazaki Y, Iwata J, Yamada K, Tomiya H, Takeda K, Inagaki Y. Effects of the new calcium antagonist efonidipine hydrochloride on resting and exercise hemodynamics in patients with stable effort angina. Arzneimittelforschung. 1996 Sep;46(9):861-7.
  30. Jump up to:a b Saruta T. Current status of calcium antagonists in Japan. Am J Cardiol. 1998;82:32R-34R.
  31. Jump up^ Okayama S, Imagawa K, Naya N, Iwama H, Somekawa S, Kawata H, Horii M, Nakajima T, Uemura S, Saito Y. Blocking T-type Ca2+ channels with efonidipine decreased plasma aldosterone concentration in healthy volunteers. Hypertens Res. 2006 Jul;29(7):493-7.
  32. Jump up^ Honda M, Hayashi K, Matsuda H, Kubota E, Tokuyama H, Okubo K, Ozawa Y, Saruta T.  Divergent natriuretic action of calcium channel antagonists in mongrel dogs: renal haemodynamics as a determinant of natriuresis. Clinical Science. 2001; 101: 421–427
  33. Jump up^ Wagner C, Kramer KB, Hinder M, Kieninger M, Kurtz A. T-type and L-type calcium channel blockers exert opposite effects on renin secretion and renin gene expression in conscious rats. Br J Pharmacol. 1998;124: 579 –585. 
  34. Jump up^ Song I, KimD, Choi S, Sun M, Kim Y, Shin HS. Role of the α1g T-type calcium channel in spontaneous absence seizures in mutant mice. J Neurosci. 2004; 24: 5249–5257.
  35. Jump up^ Lory P, Bidaud I, Chemin J. T-Type calcium channels in differentiation and proliferation. Cell Calcium. 2006; 40: 135–146.
  36. Jump up^  Ando K, Ueshima K, Tanaka S, Kosugi S, Sato T, Matsuoka H, Nakao K, Fujita T. Comparison of the antialbuminuric effects of L-/N-type and L-type calcium channel blockers in hypertensive patients with diabetes and microalbuminuria: the study of assessment for kidney function by urinary microalbumin in randomized (SAKURA) trial. Int J Med Sci. 2013 Jul 30;10(9):1209-16.
  37. Jump up to:a b Hayashi K, Wakino S, Sugano N, Ozawa Y, Homma K, Saruta T. Ca2+ Channel Subtypes and Pharmacology in the Kidney. Circ Res. 2007;100:342-353.
  38. Jump up to:a b  Hayashi K, Ozawa Y, Fujiwara K, Wakino S, Kumagai H, Saruta T. Role of actions of calcium antagonists on efferent arterioles with special references to glomerular hypertension. Am J Nephrol. 2003 Jul-Aug;23(4):229-44.
  39. Jump up to:a b Tani S, Takahashi A, Nagao K, Hirayama A. Effects of the T/L-type calcium channel blocker benidipine on albuminuria and plasma aldosterone concentration. A pilot study involving switching from L-type calcium channel blockers to benidipine. Int Heart J. 2014;55(6):519-25
  40. Jump up^  Li M.  Role of T-Type Ca2+ Channels in Basal Insulin Release. T-type Calcium Channels in Basic and Clinical Science. Springer Vienna. 2015; 137-150. 
Efonidipine
Efonidipine structure.svg
Clinical data
Trade names Landel (ランデル)
AHFS/Drugs.com International Drug Names
Routes of
administration
Oral
ATC code
  • none
Legal status
Legal status
  • In general: ℞ (Prescription only)
Identifiers
CAS Number
PubChem CID
ChemSpider
UNII
ChEMBL
Chemical and physical data
Formula C34H38N3O7P
Molar mass 631.65 g/mol
3D model (JSmol)
Title: Efonidipine
CAS Registry Number: 111011-63-3
CAS Name: 5-(5,5-Dimethyl-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylic acid 2-[phenyl(phenylmethyl)amino]ethyl ester, P-oxide
Additional Names: 2-(N-benzylanilino)ethyl(±)-1,4-dihydro-2,6-dimethyl-4-(m-nitrophenyl)-5-phosphononicotinate, cyclic 2,2-dimethyltrimethylene ester
Molecular Formula: C34H38N3O7P
Molecular Weight: 631.66
Percent Composition: C 64.65%, H 6.06%, N 6.65%, O 17.73%, P 4.90%
Literature References: Dihydropyridine calcium channel blocker. Prepn: K. Seto et al., WO 8704439idem et al., US 4885284(1987, 1989 both to Nissan); and crystal structure: R. Sakoda et al., Chem. Pharm. Bull. 40, 2362 (1992). Stereoselective synthesis of enantiomers and crystal structure of (S)-form: idem et al., ibid. 2377. Pharmacology: C. Shudo et al., J. Pharm. Pharmacol. 45,525 (1993). Mechanism of action study: T. Yamashita et al., Jpn. J. Pharmacol. 57, 337 (1991). Clinical study: T. Saito et al., Curr. Ther. Res. 52, 113 (1992).
Properties: Crystals from ethyl acetate, mp 169-170° (Sakoda); also reported as mp 155-156° (Seto).
Melting point: mp 169-170° (Sakoda); mp 155-156° (Seto)
Derivative Type: Hydrochloride
CAS Registry Number: 111011-53-1
Molecular Formula: C34H38N3O7P.HCl
Molecular Weight: 668.12
Percent Composition: C 61.12%, H 5.88%, N 6.29%, O 16.76%, P 4.64%, Cl 5.31%
Properties: LD50 in mice (mg/kg): >600 orally (Seto).
Toxicity data: LD50 in mice (mg/kg): >600 orally (Seto)
Derivative Type: Hydrochloride ethanol
CAS Registry Number: 111011-76-8
Manufacturers’ Codes: NZ-105
Trademarks: Landel (Zeria)
Molecular Formula: C34H38N3O7P.C2H5OH.HCl
Molecular Weight: 714.18
Percent Composition: C 60.54%, H 6.35%, N 5.88%, O 17.92%, P 4.34%, Cl 4.96%
Properties: Yellow crystals from aq ethanol, mp 151° (dec).
Melting point: mp 151° (dec)
Derivative Type: (S)- or (R)-Form
Properties: Pale yellow crystals from ethanol, mp 190-192°. [a]D25 + or -7.0° resp (c = 0.50 in chloroform).
Melting point: mp 190-192°
Optical Rotation: [a]D25 + or -7.0° resp (c = 0.50 in chloroform)

(R)-base

  • Formula:C34H38N3O7P
  • MW:631.67 g/mol
  • CAS-RN:128194-13-8

(S)-base

  • Formula:C34H38N3O7P
  • MW:631.67 g/mol
  • CAS-RN:128194-12-7
Therap-Cat: Antihypertensive.
Keywords: Antihypertensive; Dihydropyridine Derivatives; Calcium Channel Blocker; Dihydropyridine Derivatives.

///////////Efonidipine, エホニジピン, IND 2017, Landel , NZ 105, Efonidipine Hydrochloride Ethanolate

CC1=C(C(C(=C(N1)C)P2(=O)OCC(CO2)(C)C)C3=CC(=CC=C3)[N+](=O)[O-])C(=O)OCCN(CC4=CC=CC=C4)C5=CC=CC=C5

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