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Unoprostone
Unoprostone
- Molecular FormulaC22H38O5
- Average mass382.534 Da
Unoprostone (INN) is a prostaglandin analogue. Its isopropyl ester, unoprostone isopropyl, was marketed under the trade name Rescula for the management of open-angle glaucoma and ocular hypertension, but is now discontinued in the US.[1]
Unoprostone isopropyl is a prostaglandin analogue. Ophthalmic Solution 0.15% is a synthetic docosanoid. Unoprostone isopropyl has the chemical name isopropyl (+)-(Z)-7-[(1R,2R,3R,5S)-3,5 dihydroxy-2-(3-oxodecyl)cyclopentyl]-5-heptenoate. The main indication of Unoprostane is treatment of glucoma.
This compound can be prepared by two different ways: 1) The reaction of 1-benzyl-4-(hydroxymethyl)pyrrolidin-2-one (I) with SOCl2 in refluxing dichloromethane gives 1-benzyl-4-(chloromethyl)pyrrolidin-2-one (II), which is condensed with potassium phthalimide (III) in DMF yielding 1-benzyl-4-(phthalimidomethyl)pyrrolidin-2-one (IV). Finally, this compound is treated with hydrazine in ethanol and neutralized with fumaric acid. 2) The dehydration of 1-benzyl-2-oxo-pyrrolidine-4-carboxamide (V) with POCl3 in hot DMF gives 1-benzyl-4-cyanopyrrolidine-2-one (VI), which is reduced with H2 and RaNi in methanol – NH3 and neutralized with fumaric acid. EP 0289349; JP 1989151552; US 5001153; US 5106869
syn 2
The condensation of dimethyl methylphosphonate (I) with ethyl octanoate (II) by means of butyllithium in THF gives dimethyl 2-oxononylphosphonate (III), which is condensed with the protected aldehyde (IV) by means of NaH in THF, yielding the unsaturated ketone (V). The hydrogenation of (V) with H2 over Pd/C in ethyl acetate affords the corresponding saturated ketone (VI), which is treated with ethylene glycol and p-toluenesulfonic acid to give the cyclic ketal (VII). The mild hydrolysis of (VII) with K2CO3 and acetic acid gives the alcohol derivative (VIII); the reduction of the lactone group of (VIII) with dibutylaluminum hydride in toluene affords the lactol (IX), which is condensed with (4-carboxybutyl)triphenylphosphonium bromide (X) by means of NaH in DMSO yielding the protected prostaglandin (XI). Esterification of (XI) with isopropyl iodide and DBU in acetonitrile gives the precursor (XII), which is finally deprotected with acetic acid in THF – water.
References
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| Clinical data | |
|---|---|
| Trade names | Rescula |
| AHFS/Drugs.com | Micromedex Detailed Consumer Information |
| Pregnancy category |
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| Routes of administration |
Topical (eye drops) |
| ATC code | |
| Legal status | |
| Legal status |
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| Pharmacokinetic data | |
| Elimination half-life | 14 min |
| Excretion | Renal |
| Identifiers | |
| CAS Number |
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| PubChem CID | |
| DrugBank | |
| ChemSpider | |
| UNII | |
| KEGG | |
| ChEMBL | |
| ECHA InfoCard | 100.227.145  |
| Chemical and physical data | |
| Formula | C22H38O5 |
| Molar mass | 382.534 g/mol g·mol−1 |
| 3D model (JSmol) | |
///////////////Antiglaucoma, ocular hypertension, UF-021, Unoprostone
CCCCCCCC(=O)CC[C@H]1[C@H](O)C[C@H](O)[C@@H]1C\C=C/CCCC(O)=O
CCCCCCCC(=O)CC[C@H]1[C@H](O)C[C@H](O)[C@@H]1C\C=C/CCCC(=O)OC(C)C
HEC-68498
HEC-68498, CT-365
CAS 1621718-37-3
N-[5-(3-Cyanopyrazolo[1,5-a]pyridin-5-yl)-2-methoxy-3-pyridinyl]-2,4-difluorobenzenesulfonamide
HEC Pharm , Calitor Sciences Llc; Sunshine Lake Pharma Co Ltd
PHASE 1, idiopathic pulmonary fibrosis and solid tumors
Phosphoinositide 3-kinase inhibitor; mTOR inhibitor
- Originator HEC Pharm
- Developer HEC Pharm; Sunshine Lake Pharma
- Class Anti-inflammatories; Antifibrotics; Isoenzymes
- Mechanism of Action 1 Phosphatidylinositol 3 kinase inhibitors; MTOR protein inhibitors
- Phase I Idiopathic pulmonary fibrosis
- 22 May 2018 Phase-I clinical trials in Idiopathic pulmonary fibrosis in USA (PO) (NCT03502902)
- 24 Apr 2018 Sunshine Lake Pharma in collaboration with Covance plans a phase I trial for Idiopathic pulmonary fibrosis (In volunteers) in China , (NCT03502902)
- 19 Apr 2018 Preclinical trials in Idiopathic pulmonary fibrosis in China (PO)
- US 20140234254
- CN 103965199
CN 103965199
CN 103965199
Sunshine Lake Pharma , a subsidiary of HEC Pharm is developing an oral capsule formulation of HEC-68498 (phase 1, in July 2019) sodium salt, a dual inhibitor of phosphoinositide-3 kinase and the mTOR pathway, for the treatment of idiopathic pulmonary fibrosis and solid tumors
HEC 68498 is an oral inhibitor of phosphatidylinositol 3-kinase (PI3K) and mammalian target of rapamycin in clinical development at HEC Pharm for the treatment of idiopathic pulmonary fibrosis. A phase I trial is under way in healthy volunteers.
The phosphoinositide 3-kinases (PI3 kinases or PI3Ks), a family of lipid kinases, have been found to play key regulatory roles in many cellular processes including cell survival, proliferation and differentiation. The PI3K enzymes consist of three classes with variable primary structure, function and substrate specificity. Class I PI3Ks consist of heterodimers of regulatory and catalytic subunits, and are subdivided into 1A and 1B based on their mode of activation. Class 1A PI3Ks are activated by various cell surface tyrosine kinases, and consist of the catalytic pl lO and regulatory p85 subunits. The three known isoforms of Class 1A pl lO are pl lOot, rΐ ΐqb, and rΐ ΐqd, which all contain an amino terminal regulatory interacting region (which interfaces with p85), a Ras binding domain, and a carboxy terminal catalytic domain. Class IB PI3Ks consist of the catalytic (pl lOy) and regulatory (p 101 ) subunits and are activated by G-protein coupled receptors. (“Small-molecule inhibitors of the PI3K signaling network” Future Med. Chem ., 2011, 3, 5, 549-565).
[0004] As major effectors downstream of receptor tyrosine kinases (RTKs) and G protein-coupled receptors (GPCRs), PI3Ks transduce signals from various growth factors and cytokines into intracellular massages by generating phospholipids, which activate the serine-threonine protein kinase ART (also known as protein kinase B (PKB)) and other downstream effector pathways. The tumor suppressor or PTEN (phosphatase and tensin
homologue) is the most important negative regulator of the PI3K signaling pathway. (“Status of PBK/Akt/mTOR Pathway Inhibitors in Lymphoma.” Clin Lymphoma, Myeloma Leuk , 2014, 14(5), 335-342.)
[0005] The signaling network defined by phosphoinositide 3-kinases (PI3Ks), AKT and mammalian target of rapamycin (mTOR) controls most hallmarks of cancer, including cell cycle, survival, metabolism, motility and genomic instability. The pathway also contributes to cancer promoting aspects of the tumor environment, such as angiogenesis and inflammatory cell recruitment. The lipid second messenger produced by PI3K enzymes, phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3; also known as PIP3), is constitutively elevated in most cancer cells and recruits cytoplasmic proteins to membrane-localized‘onco’ signal osomes.
[0006] Cancer genetic studies suggest that the PI3K pathway is the most frequently altered pathway in human tumors: the PIK3CA gene (encoding the PI3K catalytic isoform pl lOa) is the second most frequently mutated oncogene, and PTEN (encoding phosphatase and tensin homolog, the major PtdIns(3,4,5)P3 phosphatase) is among the most frequently mutated tumor suppressor genes. In accord, a recent genomic study of head and neck cancer found the PI3K pathway to be the most frequently mutated. Indeed, even in cancer cells expressing normal PI3K and PTEN genes, other lesions are present that activate the PI3K signaling network (that is, activated tyrosine kinases, RAS and AKT, etc ). As a net result of these anomalies, the PI3K pathway is activated, mutated or amplified in many malignancies, including in ovarian cancer (Campbell et al., Cancer Res., 2004, 64, 7678-7681; Levine et al., Clin. Cancer Res., 2005, 11, 2875-2878; Wang et al., Hum. Mutat., 2005, 25, 322; Lee et al., Gynecol. Oncol. ,2005, 97, 26-34), cervical cancer, breast cancer (Bachman et al.,· Cancer Biol., Ther, 2004, 3, 772-775; Levine et al., supra; Li et al., Breast Cancer Res. Treat., 2006, 96, 91-95; Saal et al., Cancer Res., 2005, 65, 2554-2559; Samuels and Velculescu, Cell Cycle, 2004, 3, 1221-1224), colorectal cancer (Samuels et al., Science, 2004, 304, 554; Velho et al., Eur. J. Cancer, 2005, 41, 1649-1654), endometrial cancer (Oda et al ., Cancer Res., 2005, 65, 10669-10673), gastric carcinomas (Byun et al., M. J. Cancer, 2003 , 104, 318-327; Li et al., supra; Velho et al., supra; Lee et al., Oncogene, 2005 , 24, 1477-1480), hepatocellular carcinoma (Lee et al., id), small and non-small cell lung cancer (Tang et al., Lung Cancer 2006, 11, 181-191; Massion et al , Am. J. Respir. Crit. Care Med., 2004, 170, 1088-1094), thyroid carcinoma (Wu et al., J. Clin. Endocrinol. Metab., 2005, 90, 4688-4693),
acute myelogenous leukemia (AML) (Sujobert et al., Blood, 1997, 106, 1063-1066), chronic myelogenous leukemia (CML) (Hickey et al., J. Biol. Chem ., 2006, 281, 2441-2450), glioblastomas (Hartmann et al. Jlcta Neuropathol (Bert ), 2005, 109, 639-642; Samuels et al., supra), Hodgkin and non-Hodgkin lymphomas (“PI3K and cancer: lessons, challenges and opportunities”, Nature Reviews Drug Discovery., 2014, 13, 140).
[0007] The PI3K pathway is hyperactivated in most cancers, yet the capacity of PI3K inhibitors to induce tumor cell death is limited. The efficacy of PI3K inhibition can also derive from interference with the cancer cells’ ability to respond to stromal signals, as illustrated by the approved PI3K5 inhibitor idelalisib in B-cell malignancies. Inhibition of the leukocyte-enriched PI3K5 or RI3Kg may unleash antitumor T-cell responses by inhibiting regulatory T cells and immune-suppressive myeloid cells. Moreover, tumor angiogenesis may be targeted by PI3K inhibitors to enhance cancer therapy. (“Targeting PI3K in Cancer: Impact on Tumor Cells, Their Protective Stroma, Angiogenesis, and Immunotherapy”, Cancer Discov., 2016, 6(10), 1090-1105.)
[0008] mTOR is a highly conserved serine-threonine kinase with lipid kinase activity and participitates as an effector in the PI3K/AKT pathway. mTOR exists in two distinct complexes, mTORCl and mTORC2, and plays an important role in cell proliferation by monitoring nutrient avaliability and cellular energy levels. The downstream targets of mTORCl are ribosomal protein S6 kinase 1 and eukaryotic translation initiation factor 4E-binding protein 1, both of which are crucial to the regulation of protein synthesis. (“Present and future of PI3K pathway inhibition in cancer: perspectives and limitations”, Current Med. Chem., 2011, 18, 2647-2685).
[0009] Knowledge about consequences of dysregulated mTOR signaling for tumorigenesis comes mostly from studies of pharmacologically disruption of mTOR by repamycin and its analogues such as temsirolimus (CCI-779) and everolimus (RADOOl).Rapamycin was found to inhibit mTOR and thereby induce G1 arrest and apoptosis. The mechanism of rapamycin growth inhibition was found to be related to formation of complexes of rapamycin with FK-binding protein 12 (FKBP-12). These complexes then bound with high affinity to mTOR, preventing activation and resulting in inhibition of protein translation and cell growth. Cellular effects of mTOR inhibition are even more pronounced in cells that have concomitant inactivation of PTEN. Antitumor activity of rapamycin was subsequently identified, and a number of rapamycin analogues such as temsirolimus and everolimus have been approved by the US Food and Drug
Administration for the treatment certain types of cancer.
[0010] Fibrosis is the formation of excess fibrous connective tissue in an organ or tissue in a reparative or reactive process. Examples of fibrosis include, but are not limited to pulmonary fibrosis, liver fibrosis, dermal fibrosis, and renal fibrosis. Pulmonary fibrosis, also called idiopathic pulmonary fibrosis (IPF), interstitial diffuse pulmonary fibrosis, inflammatory pulmonary fibrosis, or fibrosing alveolitis, is a lung disorder and a heterogeneous group of conditions characterized by abnormal formation of fibrous tissue between alveoli caused by alveolitis comprising cellular infiltration into the alveolar septae with resulting fibrosis. The effects of IPF are chronic, progressive, and often fatal.
[0011] The clinical course of IPF is variable and largely unpredictable. IPF is ultimately fatal, with historical data suggesting a median survival time of 2-3 years from diagnosis. A decline in forced vital capacity (FVC) is indicative of disease progression in patients with IPF and change in FVC is the most commonly used endpoint in clinical trials. A decline in FVC of 5% or 10% of the predicted value over 6-12 months has been associated with increased mortality in patients with IPF.
[0012] Our understanding of the pathogenesis of IPF has evolved from that of a predominantly inflammatory disease to one driven by a complex interplay of repeated epithelial cell damage and aberrant wound healing, involving fibroblast recruitment, proliferation and differentiation, and culminating in excess deposition of extracellular matrix. This shift in knowledge prompted a change in the type of compounds being investigated as potential therapies, with those targeted at specific pathways in the development and progression of fibrosis becoming the focus.
[0013] In patients with IPF, the mechanisms whereby PI3K/mTOR inhibitors act may involve inhibition of kinases such as PI3Ks and mTOR. This results in inactivation of cellular receptors for mediators involved in the development of pulmonary fibrosis. As a result, fibroblast proliferation is inhibited and extracellular matrix deposition is reduced. (“Update on diagnosis and treatment of idiopathic pulmonary fibrosis”, J Bras Pneumol. 2015, 41(5), 454-466.)
[0014] Accordingly, small-molecule compounds that specially inhibit, regulate and/or modulate the signal transduction of kinases, particularly including PI3K and mTOR as described above, are desirable as a means to prevent, manage, or treat proliferative disorders and fibrosis, particular idiopathic pulmonary fibrosis in a patient. One such small-molecule is A-(5-(3-cyanopyrazolo[l,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfon-amide, which has the chemical structure as shown in the following:
[0015] WO 2014130375A1 described the synthesis of N-(5 -(‘3 -cyanopyrazol o [l,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfonamide (Example 3) and also disclosed the therapeutic activity of this molecule in inhibiting, regulating and modulating the signal transduction of protein kinases.
[0016] Different salts and solid state forms of an active pharmaceutical ingredient may possess different properties. Such variations in the properties of different salts and solid state forms may provide a basis for improving formulation, for example, by facilitating better processing or handling characteristics, improving the dissolution profile, stability (polymorph as well as chemical stability) and shelf-life. These variations in the properties of different salts and solid state forms may also provide improvements to the final dosage form, for example, if they serve to improve bioavailability. Different salts and solid state forms of an active pharmaceutical ingredient may also give rise to a variety of polymorphs or crystalline forms, which may in turn provide additional opportunities to assess variations in the properties and characteristics of a solid active pharmaceutical ingredient.
Different salts and solid state forms of /V-(5-(3-cyanopyrazolo[l,5- ]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfonamide are described herein.
PATENT
WO2014130375 ,
https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2014130375
claiming new pyrazolo[1,5-a]pyridine derivatives are PI3K and mTOR inhibitors, useful for treating proliferative diseases
Example 3 N-(5-(3-cyanopyrazolo[1,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfonamide
Step 1) 5-bromopyrazolo[1,5-a]pyridine
[196] A solution of ethyl 5-bromopyrazolo[1,5-a]pyridine-3-carboxylate (240
mmol) in 40% H2SO4 (12 mL) was stirred at 100 °C for 4 hours, then cooled to rt, and neutralized to pH=7 with aq. NaOH (6 M) in ice bath. The resulted mixture was extracted with DCM (25 mL x 2). The combined organic phases were dried over anhydrous Na2SO4 and concentrated in vacuo to give the title compound as a light yellow solid (175 mg, 99.5%).
MS (ESI, pos. ion) m/z: 196.9 [M+H]+.
Step 2) 5-bromopyrazolo[1,5-a]pyridine-3-carbaldehyde
[197] To a solution of 5-bromopyrazolo[1,5-a]pyridine (175 mg, 0.89 mmol) in DCM (6 mL) was added (chloromethylene)dimethyliminium chloride (632 mg, 3.56 mmol). The reaction was stirred at 44 °C overnight, and concentrated in vacuo. The residue was dissolved in saturated NaHCO3 aqueous solution (25 mL) and the resulted mixture was then extracted with EtOAc (25 mL x 3). The combined organic phases were dried over anhydrous Na2SO4 and concentrated in vacuo to give the title compound as a light yellow solid (225 mg, 100%).
MS (ESI, pos. ion) m/z: 225.0 [M+H]+.
Step 3) (E)-5-bromopyrazolo[1,5-a]pyridine-3-carbaldehyde oxime
[198] To a suspension of 5-bromopyrazolo[1,5-a]pyridine-3-carbaldehyde (225 mg, 1 mmol) in EtOH (10 mL) and H2O (5 mL) was added hydroxylamine hydrochloride (104 mg, 1.5 mmol). The reaction was stirred at 85 °C for 2 hours, then cooled to rt, and concentrated in vacuo. The residue was adjusted to pH=7 with saturated NaHCO3 aqueous solution. The resulted mixture was then filtered and the filter cake was dried in vacuo to give title compound as a yellow solid (240 mg, 99%).
MS (ESI, pos. ion) m/z: 240.0 [M+H]+.
Step 4) 5-bromopyrazolo[1,5-a]pyridine-3-carbonitrile
[199] A solution of (E)-5-bromopyrazolo[1,5-a]pyridine-3-carbaldehyde oxime (240 mg,
1 mmol) in Ac2O (6 mL) was stirred at 140 °C for 18 hours, then cooled to rt, and concentrated in vacuo. The residue was washed with Et2O (1 mL) to give the title compound as a yellow solid (44 mg, 22.5%).
MS (ESI, pos. ion) m/z: 222.0 [M+H]+.
Step 5) N-(5-(3-cyanopyrazolo[1,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluorobenzenesulfonamide
[200] 2,4-difluoro-N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)benzenesulfonamide (612 mg, 1.5 mmol), 5-bromopyrazolo[1,5-a]pyridine-3-carbonitrile (222 mg, 1 mmol), Pd(dppf)Cl2·CH2Cl2 (16 mg, 0.02 mmol) and Na2CO3 (85 mg, 0.8 mmol) were placed into a two-neck flask, then degassed with N2 for 3 times, and followed by adding 1,4-dioxane (5 mL) and water (1 mL). The resulted mixture was degassed with N2 for 3 times, then heated to 90 °C and stirred further for 5 hours. The mixture was cooled to rt and filtered. The filtrate was concentrated in vacuo and the residue was purified by a silica gel column chromatography (PE/EtOAc (v/v) = 1/2) to give the title compound as a light yellow solid (400 mg, 81.6%).
MS (ESI, pos. ion) m/z: 442.0 [M+H]+;
1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.37 (s, 1H), 9.02 (d, J = 7.2 Hz, 1H), 8.67 (s, 1H), 8.60 (d, J = 2.2 Hz, 1H), 8.26-8.16 (m, 2H), 7.82-7.72 (m, 1H), 7.57 (dd, J = 13.2, 5.8 Hz, 2H), 7.21 (t, J= 8.5 Hz, 1H), 3.67 (s, 3H).
PATENT
WO-2019125967
The invention relates to salts of pyrazolo[l,5-a]pyridine derivatives and use thereof, specifically relates to salt of /V-(5-(3-cyanopyrazolo[l,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl) -2,4-difluorobenzenesulfonamide (compound of formula (I)) and use thereof, further relates to composition containing said salts above. The salts or the composition can be used to inhibit/modulate protein kinases, further prevent, manage or treat proliferative disorders or pulmonary fibrosis in a patient.
Amorphous form of mono-sodium salt of HEC-68498 , useful for treating a proliferative disorder or pulmonary fibrosis.
The invention is further illustrated by the following examples, which are not be construed as limiting the invention in scope.
[00108] /V-(5-(3-cyanopyrazolo[l,5-a]pyridin-5-yl)-2-methoxypyridin-3-yl)-2,4-difluoroben zenesulfonamide can be prepared according to the synthetic method of example 3 disclosed in WO2014130375 Al.
//////////////HEC-68498, HEC 68498, HEC68498, HEC Pharm , Calitor Sciences, Sunshine Lake Pharma, PHASE 1, proliferative disorder, pulmonary fibrosis, idiopathic pulmonary fibrosis, solid tumors, CT-365 , CT 365 , CT365
Fc1ccc(c(F)c1)S(=O)(=O)Nc2cc(cnc2OC)c3ccn4ncc(C#N)c4c3
GNE-0877
GNE-0877
Maybe DNL-151 ?
CAS 1374828-69-9
Chemical Formula: C14H16F3N7
Molecular Weight: 339.31895
2-methyl-2-(3-methyl-4-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-pyrazol-1-yl)propanenitrile
Denali Therapeutics Inc, useful for treating Alzheimer’s disease, breast tumor, type I diabetes mellitus and Crohn’s disease
GNE-0877 is a highly potent and selective LRRK2 inhibitor. Leucine-rich repeat kinase 2 (LRRK2) has drawn significant interest in the neuroscience research community because it is one of the most compelling targets for a potential disease-modifying Parkinson’s disease therapy.
- Developer Denali Therapeutics Inc
- Class Antiparkinsonians; Small molecules
- Mechanism of Action LRRK2 protein inhibitors
- Phase I Parkinson’s disease
- 20 Dec 2017 Denali Therapeutics plans clinical studies for Parkinson’s disease
- 13 Nov 2017 Phase-I clinical trials in Parkinson’s disease (In volunteers) in Netherlands (unspecified route)
- 13 Nov 2017 Preclinical trials in Parkinson’s disease in USA (unspecified route) before November 2017
Denali Therapeutics is developing DNL-151 (phase 1, in July 2019), a lead from a program of small-molecule inhibitors of LRRK2 originally licensed from Genentech, for the treatment of Parkinson’s disease.
Leucine-rich repeat kinase 2 (LRRK2) is a complex signaling protein that is a key therapeutic target, particularly in Parkinson’s disease (PD). Combined genetic and biochemical evidence supports a hypothesis in which the LRRK2 kinase function is causally involved in the pathogenesis of sporadic and familial forms of PD, and therefore that LRRK2 kinase inhibitors could be useful for treatment (Christensen, K.V. (2017) Progress in medicinal chemistry 56:37-80). Inhibition of the kinase activity of LRRK2 is under investigation as a possible treatment for Parkinson’s disease (Fuji, R.N. et al (2015) Science Translational Medicine 7(273):ral5;
Taymans, J.M. et al (2016) Current Neuropharmacology 14(3):214-225). A group of LRRK2 kinase inhibitors have been studied (Estrada, A.A. et al (2015) Jour. Med. Chem. 58(17): 6733-6746; Estrada, A.A. et al (2013) Jour. Med. Chem. 57:921-936; Chen, H. et al (2012) Jour. Med. Chem. 55:5536-5545; Estrada, A.A. et al (2015) Jour. Med. Chem. 58:6733-6746; US 8354420; US 8569281; US8791130; US 8796296; US 8802674; US 8809331; US 8815882; US 9145402; US 9212173; US 9212186; WO 2011/151360; WO 2012/062783; and WO 2013/079493.
PATENT
WO2012062783 , assigned to Hoffmann-La Roche , but naming inventors specifically associated with both Genentech and BioFocus (which had an agreement with Genentech for drug discovery programs); the compound was also later identified in J.Med.Chem (57(3), 921-936, 2014) in an article from these two companies, with the lab code GNE-0877. So while this represents the first application in the name of Denali Therapeutics Inc that focuses on this compound, it is likely that it provides the structure of DNL-151 , a lead from a program of small-molecule inhibitors of leucine-rich repeat kinase 2 (LRRK2) originally licensed from Genentech, being developed for the oral treatment of Parkinson’s disease, and which had begun phase I trials by December 2017 (when this application was lodged).
PATENT
WO2019104086 ,
https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2019104086
claiming novel crystalline and amorphous forms of pyrimidinylamino-pyrazole compound, useful for treating Alzheimer’s disease, breast tumor, type I diabetes mellitus and Crohn’s disease.
Novel crystalline and amorphous forms of 2-methyl-2-(3-methyl-4-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-pyrazol-1-yl)propanenitrile (which is substantially pure form) and their anhydrous and solvates such as cyclohexanol solvate (designated as Forms B-D), processes for their preparation and compositions comprising them are claimed. The compound is disclosed to be leucine rich serine threonine kinase 2 inhibitor, useful for treating Gaucher disease, Alzheimer’s disease, motor neurone disease, Parkinson’s disease, prostate tumor, Lewy body dementia, mild cognitive impairment, breast tumor, type I diabetes mellitus and Crohn’s disease.
The present disclosure relates to crystalline polymorph or amorphous forms of a pyrimidinylamino-pyrazole kinase inhibitor, referred to herein as the Formula I compound and having the structure:
FORMULA I COMPOUND
The present disclosure includes polymorphs and amorphous forms of Formula I compound, (CAS Registry Number 1374828-69-9), having the structure:
and named as: 2-methyl-2-(3-methyl-4-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-lH-pyrazol-l-yl)propanenitrile (WO 2012/062783; US 8815882; US 2012/0157427, each of which are incorporated by reference). As used herein, the Formula I compound includes tautomers, and pharmaceutically acceptable salts or cocrystals thereof. The Formula I compound is the API (Active Pharmaceutical Ingredient) in formulations for use in the treatment of neurodegenerative and other disorders, with pKa when protonated calculated at 6.7 and 2.1.
CRYSTALLIZATION
Initial polymorph screening experiments were performed using a variety of
crystallization or solid transition methods, including: anti-solvent addition, reverse anti-solvent addition, slow evaporation, slow cooling, slurry at room temperature (RT), slurry at 50 °C, solid vapor diffusion, liquid vapor diffusion, and polymer induced crystallization. By all these methods, the Form A crystal type was identified. Polarized light microscopy (PLM) images of Form A obtained from various polymorph screening methods were collected (Example 5).
Particles obtained via anti-solvent addition showed small size of about 20 to 50 microns (pm) diameter while slow evaporation, slow cooling (except for THF/isooctane), liquid vapor diffusion and polymer-induced crystallization resulted in particles with larger size. Adding isooctane into a dichloromethane (DCM) solution of the Formula I compound produced particles with the most uniform size. Crude Formula I compound crystallized from THF///-heptane and then was micronized. A crystallization procedure was developed to control particle size.
A total of four crystal forms (Forms A, B, C, and D) and an amorphous form E of Formula I compound were prepared, including 3 anhydrates (Form A, C, and D) and one solvate (Form B). Slurry competition experiments indicated that Form D was thermodynamically more stable when the water activity aw< 0.2 at RT, while Form C was more stable when aw> 0.5 at RT. The 24 hrs solubility evaluation showed the solubility of Form A, C and D in FLO at RT was 0.18, 0.14 and 0.11 mg/mL, respectively. DVS (dynamic vapor sorption) results indicated that Form A and D were non-hygroscopic as defined by less than 0.1% reversible water intake in DVS, while Form C was slightly hygroscopic. Certain characterization data and observations of the crystal forms are shown in Table 1.
Table 1 Characterization summary for crystal forms of Formula I compound
Differential Scanning Calorimetry (DSC) analysis of Forms A and C showed that Form C had higher melting point and higher heat of fusion (Table 1), suggesting that the two forms are monotropic with Form C being the more stable form. Competitive slurry experiments with 1 : 1 Form A and C in a variety of solvents always produced Form C confirming that Form C was
more stable than Form A. In accordance with this, Form C was produced even when the crystallization batch was seeded with seeds of Form A.
PATENT
WO-2019126383
Methods of making leucine-rich repeat kinase 2 (LRRK2)-inhibiting, pyrimidinyl-4-aminopyrazole compounds (eg 2-methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)- lH-pyrazol-1-yl)propanenitrile), useful for treating LRRK2 mediated diseases such as Parkinson’s disease.
Example 1 Preparation of 2-(4-amino-3 -methyl- liT-pyrazol-l -yl)-2-methylpropanamide 5a
4a 5a
To a 20-L reactor containing dimethyl formamide (4.5 L) was charged 5-methyl-4-nitro-lH-pyrazole la (1.5 kg, 1.0 equiv). The solution was cooled to 0 °C and charged with finely ground K2CO3 (2.45 kg, 1.5 equiv) in three portions over ~l h. Methyl 2-bromo-2-methylpropanoate (3.2 kg, 1.5 equiv) was added dropwise to the mixture and then was allowed to warm to ~25 °C. The reaction mixture was maintained for 16 h and then quenched with water (15 L) and product was extracted with ethyl acetate. The combined organic layer was washed with water, and then with a brine. The organic layer was dried over anhydrous Na2S04, filtered, and concentrated under reduced pressure to give a light yellow solid. The crude product was purified by crystallization with petroleum ether (15 L), filtered, and dried to give methyl 2-m ethyl -2-(3 -methyl -4-nitro- l//-pyrazol- l -yl)propanoate 3a (2.25 kg, >99% purity by HPLC, 84 % yield) as an off-white solid. ¾ NMR (400 MHz, CDCb) 8.28 (s, 1H), 3.74 (s, 3H), 2.53 (s, 3H), 1.85 (s, 6H).
Methanol (23 L) and 2-methyl-2-(3-methyl-4-nitro-lif-pyrazol-l-yl)propanoate 3a (2.25 kg, 1.0 equiv) were charged into a 50-L reactor and cooled to approximately -20 °C. Ammonia gas was purged over a period of 5 h and then the reaction mixture warmed to 25 °C. After 16 h, the reaction mixture was concentrated under reduced pressure (~50 °C) to give the crude product. Ethyl acetate (23 L) was charged and the solution agitated in the presence of charcoal (0.1 w/w) and Celite® (0.1 w/w) at 45 °C. The mixture was filtered and concentrated under reduced pressure, and then the solid was slurried in methyl tert-butyl ether (MTBE, 11.3 L) at RT for 2 h. Filtration and drying at ~45 °C gave 2-m ethyl -2-(3 -m ethyl -4-ni tro- 1 //-pyrazol – 1 -yl)propanamide 4a (1.94 kg, >99% purity by HPLC, 92% yield).
Methanol (5 L) and 2-m ethyl-2-(3 -methyl -4-nitro-lif-pyrazol-l-yl)propanamide 4a (0.5 kg) were charged into a 10-L autoclave under nitrogen atmosphere, followed by slow addition of 10 % (50% wet) Pd/C (50 g). Hydrogen was charged (8.0 kg pressure/l 13 psi) and the reaction mixture agitated at 25 °C until complete. The mixture was filtered, concentrated under reduced
pressure and then slurried in MTBE (2.5 L) for 2 h at 25 °C. Filtration and drying under reduced pressure (45 °C) gave 2-(4-amino-3-methyl- l//-pyrazol- l -yl)-2-methyl propanamide 5a (0.43 kg, >99% purity by HPLC, 99% yield).
Example 2 Preparation of 2-(4-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-lH-pyrazol-l-yl)-2-methylpropanamide 7a
DCM
Into a first reactor was charged /-BuOH (or alternatively 2-propanol) (15.5 vol) and 2-(4-amino-3 -methyl- li7-pyrazol-l-yl)-2-methylpropanamide 5a (15 kg), followed by zinc chloride (13.5 kg, 1.2 equiv) at room temperature and the suspension agitated ~2 h. Into a second reactor was charged dichloromethane (DCM, 26.6 vol) and 2,4-dichloro-5-trifluoromethyl pyrimidine 6a (19.6 kg, 1.1 equiv) and then cooled to 0 °C. The contents from first reactor were added portion-wise to the second reactor. After addition, the reaction mixture was agitated at 0 °C for ~l h and then Et3N (9.2 kg, 1.1 equiv) was slowly charged. After agitation for 1 h, the temperature was increased to 25 °C and monitored for consumption of starting material. The reaction mixture was quenched with 5% aqueous NaHCO, and then filtered over Celite®. The DCM layer was removed and the aqueous layer was back-extracted with DCM (3x). The combined organics were washed with water, dried (Na2S04), and concentrated. Methanol (2.5 vol) was charged and the solution was heated to reflux for 1 h, then cooled to 0 °C. After 1 h, the solids were filtered and dried under reduced pressure to give 2-(4-((4-chloro-5-(tri fluoromethyl)pyri mi din-2-yl)amino)-3 -methyl – l//-pyrazol- l -yl)-2-methyl propanamide 7a
(31.2 kg (wet weight)). 1H NMR (600 MHz, DMSO-de) 10.05 (br. s., 1H), 8.71 (d, J= 11 Hz, 1H), 7.95 (app. d, 1H), 7.18 (br. s., 1H), 6.78 (br. s., 1H), 2.14 (s, 3H), 1.67 (s, 6H).
Example 3 Preparation of 2-methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)- lH-pyrazol- 1 -yl)propanamide 8a
A reactor was charged with anhydrous tetrahydrofuran (THF, 10 vol) and 2-(4-((4-chloro-5-(trifl uoromethyl )pyrimi din-2-yl)amino)-3 -methyl – l //-pyrazol- l -yl)-2-methylpropanamide 7a (21 kg) at room temperature with agitation. A solution of 2M
methylamine in THF (3.6 vol) was slowly charged to the reactor at 25 °C and maintained for ~3 h. The reaction mixture was diluted with 0.5 w/w aqueous sodium bicarbonate solution (10 w/w), and extracted with ethyl acetate (EtOAc, 4.5 w/w). The aqueous layer was extracted with EtOAc (4x), the organics were combined and then washed with H20 (7 w/w). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. «-Heptane (3 w/v) was added to the residue, agitated, filtered and dried under reduced pressure to give 2-m ethyl -2-(3 -methyl -4-((4-(methyl ami no)-5-(trifl uoromethyl )pyri mi din-2-yl)amino)- l //-pyrazol-1 -yl)propanamide 8a (19.15 kg, 93% yield). ¾ NMR (600 MHz, DMSO-d6) 8.85 (m, 1H), 8.10 (s, 1H), 8.00 (m, 1H), 7.16 (br. s., 1H), 6.94 (m, 1H), 6.61 (br. s., 1H), 2.90 (d, J = 4.3 Hz, 3H), 2.18 (br. s., 3H), 1.65 (s, 6H).
Example 4 Preparation of 2-methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)- lH-pyrazol- 1 -yl)propanenitrile 9a
To a reactor was charged 2-methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifl uoromethyl )pyri mi din-2-yl)amino)- l //-pyrazol- l -yl)propan amide 8a (15 kg, 1 equiv) at room temperature followed by EtOAc (2 vol) and 6.7 vol T3P (50% w/w in EtOAc). The reaction mixture was heated to 75 °C over 1 h and then agitated for 16 h until consumption of starting material. The reaction mixture was cooled between -10 to -15 °C then added drop-wise 5N aqueous NaOH (7 vol) resulting in pH 8-9. The layers were separated and the aqueous layer back-extracted with EtOAc (2 x 4 vol). The combined organic extracts were washed with 5 %
aqueous NaHCO, solution, and then distilled to azeotropically remove water. The organics were further concentrated, charged with «-heptane (2 vol) and agitated for 1 h at room temperature. The solids were filtered, rinsed with «-heptane (0.5 vol) and then dried under vacuum (<50 °C). The dried solids were dissolved in EtOAc (1.5 vol) at 55 °C, and then «-heptane (3 vol) was slowly added followed by 5-10% of 9a seeds. To the mixture was slowly added «-heptane (7 vol) at 55 °C, agitated for 1 h, cooled to room temperature and then maintained for 16 h. The suspension was further cooled between 0-5 °C, agitated for 1 hour, filtered, and then rinsed the filter with chilled 1 :6.5 EtOAc/«-heptane (1 vol). The product was dried under vacuum at 50 °C to give 2-methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1 //-pyrazol – 1 -yl )propaneni tri 1 e 9a (9.5 kg, first crop), 67% yield). ‘H NMR (600 MHz, DMSO-d6) 8.14 (s, 1H), 8.13 (br. s., 1H), 7.12 (br. s., 1H), 5.72 (br. s, 1H), 3.00 (d, J= 4.6 Hz, 3H),
2.23 (s, 3H), 1.96 (s, 3H).
Example 5 Preparation of methyl 2-(4-amino-3-methyl-lH-pyrazol-l-yl)-2-methylpropanoate 10a
Following the procedure of Example 1, a mixture of methanol and methyl 2-methyl-2-(3-methyl-4-nitro-lH-pyrazol-l-yl)propanoate 3a (0.5 kg) was charged into an autoclave under nitrogen atmosphere, followed by slow addition of 10 % (50% wet) Pd/C. Hydrogen was charged under pressure and the reaction mixture agitated at 25 °C until complete. The mixture was filtered, concentrated under reduced pressure and then slurried in MTBE for 2 h at 25 °C. Filtration and drying under reduced pressure gave methyl 2-(4-amino-3-methyl-lH-pyrazol-l-yl)-2-methylpropanoate 10a (LC-MS, M+l=l98).
Example 6 Preparation of methyl 2-(4-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3 -methyl- lH-pyrazol- 1 -yl)-2-methylpropanoate 11a
Following the procedure of Example 2, a mixture of methyl 2-(4-amino-3-methyl-lH-pyrazol-l-yl)-2-methylpropanoate 10a and DIPEA (1.2 equiv) in /-BuOH was warmed to 80 °C. Then a solution of 2,4-dichloro-5-trifluoromethyl pyrimidine 6a in /-BuOH was added slowly drop wise at 80 °C. After 15 minutes, LCMS showed the reaction was complete, including later eluting 59.9% of product ester 11a, earlier eluting 31.8% of undesired regioisomer (ester), and no starting material 10a. After completion of reaction, the mixture was cooled to room temperature and a solid was precipitated. The solid precipitate was filtered and dried to give methyl 2-(4-((4-chloro-5-(trifluoromethyl)pyrimi din-2 -yl)amino)-3-methyl-lH-pyrazol-l-yl)-2-methylpropanoate 11a (LC-MS, M+l=378).
PAPER
J.Med.Chem (57(3), 921-936, 2014
https://pubs.acs.org/doi/full/10.1021/jm401654j
2-Methyl-2-(3-methyl-4-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propanenitrile (11)
aReagents and conditions: (a) NaH, methyl 2-bromo-2-methylpropanoate, DMF, 70%; (b) LiOH, THF-H2O, 90%; (c) (i) (COCl)2, CH2Cl2, (ii) R-NH2, THF; (d) Pd/C, H2, MeOH; (e) 26, Et3N, n-BuOH, 120 °C; (f) 26, TFA, 2-methoxyethanol, 70 °C; (g) POCl3, 90 °C, 42%.
GNE-9605
CAS № 1536200-31-3
GNE-9065 is an orally bioavailable and potent inhibitor of leucine-rich repeat kinase 2 (LRRK2; IC50 = 18.7 nM).1 It is selective for LRRK2 over 178 kinases, inhibiting only TAK1-TAB1 >50% at a concentration of 0.1 μM. GNE-9065 (10 and 50 mg/kg) inhibits LRRK2 Ser1292 autophosphorylation in BAC transgenic mice expressing human LRRK2 protein with the G2019S mutation found in families with autosomal Parkinson’s disease.
CNC1=C(C(F)(F)F)C=NC(NC2=C(Cl)N([C@H]3CCN(C4COC4)C[C@@H]3F)N=C2)=N1
N2-(5-Chloro-1-((trans)-3-fluoro-1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-N4-methyl-5-(trifluoromethyl)pyrimidine-2,4-diamine (20)
aReagents and conditions: (a) (±)-(cis)-tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate, PPh3, diisopropyl azodicarboxylate, THF; (b) TFA, DCM, 58% over two steps; (c) oxetan-3-one, DIPEA, NaBH(OAc)3, acetic acid, DCE, 85%; (d) LiHMDS then C2Cl6, THF, −78 °C, 65%; (e) iron dust, NH4Cl, EtOH, 90 °C; (f) 26, TFA, 2-methoxyethanol, 90 °C, 40%, two steps.
REFERENCES
1: Estrada AA, Chan BK, Baker-Glenn C, Beresford A, Burdick DJ, Chambers M, Chen H, Dominguez SL, Dotson J, Drummond J, Flagella M, Fuji R, Gill A, Halladay J, Harris SF, Heffron TP, Kleinheinz T, Lee DW, Pichon CE, Liu X, Lyssikatos JP, Medhurst AD, Moffat JG, Nash K, Scearce-Levie K, Sheng Z, Shore DG, Wong S, Zhang S, Zhang X, Zhu H, Sweeney ZK. Discovery of Highly Potent, Selective, and Brain-Penetrant Aminopyrazole Leucine-Rich Repeat Kinase 2 (LRRK2) Small Molecule Inhibitors. J Med Chem. 2014 Jan 15. [Epub ahead of print] PubMed PMID: 24354345.
/////////////DNL-151, DNL 151, DNL151, Alzheimer’s disease, breast tumor, type I diabetes mellitus, Crohn’s disease, phase 1, Parkinson’s disease, GNE0877, GNE 0877, GNE-0877, GNE-9605, GNE 9605, GNE9605, Genentech
CC(N1N=C(C)C(NC2=NC=C(C(F)(F)F)C(NC)=N2)=C1)(C)C#N
Nicotinamide riboside chloride
Nicotinamide riboside chloride
CAS 23111-00-4 CHLORIDE
CAS : 1341-23-7 (cation) 23111-00-4 (chloride) 445489-49-6 (Triflate)
3-Carbamoyl-1-((2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-ium chloride
Nicotinamide ribose chloride
UNII-8XM2XT8VWI
MW 290.7 g/mol
1-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]pyridin-1-ium-3-carboxamide;chloride
C1=CC(=C[N+](=C1)C2C(C(C(O2)CO)O)O)C(=O)N.[Cl-]
Nicotinamide riboside; SRT647; SRT-647; SRT 647; Nicotinamide Riboside Triflate, α/β mixture
EH-301, nicotinamide riboside chloride,AND pterostilbene,, BY Elysium Health Inc
Nicotinamide riboside, also known as NR and SRT647, is a pyridine-nucleoside form of vitamin B3 that functions as a precursor to nicotinamide adenine dinucleotide or NAD+. NR blocks degeneration of surgically severed dorsal root ganglion neurons ex vivo and protects against noise-induced hearing loss in living mice. Nicotinamide riboside prevents muscle, neural and melanocyte stem cell senescence. Increased muscular regeneration in mice has been observed after treatment with nicotinamide riboside, leading to speculation that it might improve regeneration of organs such as the liver, kidney, and heart. Nicotinamide riboside also lowers blood glucose and fatty liver in prediabetic and type 2 diabetic models while preventing the development of diabetic peripheral neuropathy. Note: Nicotinamide Riboside chloride is a α/β mixture
Nicotinamide riboside (NR) is a pyridine–nucleoside form of vitamin B3 that functions as a precursor to nicotinamide adenine dinucleotide or NAD+.[1][2]
Chemistry
While the molecular weight of nicotinamide riboside is 255.25 g/mol,[3] that of its chloride salt is 290.70 g/mol.[4][5] As such, 100 mg of nicotinamide riboside chloride provides 88 mg of nicotinamide riboside.
History
Nicotinamide riboside (NR) was first described in 1944 as a growth factor, termed Factor V, for Haemophilus influenza, a bacterium that lives in and depends on blood. Factor V, purified from blood, was shown to exist in three forms: NAD+, NMN and NR. NR was the compound that led to the most rapid growth of this bacterium.[6] Notably, H. influenza cannot grow on nicotinic acid, nicotinamide, tryptophan or aspartic acid, which were the previously known precursors of NAD+.[7]
In 2000, yeast Sir2 was shown to be an NAD+-dependent protein lysine deacetylase,[8] which led several research groups to probe yeast NAD+ metabolism for genes and enzymes that might regulate lifespan. Biosynthesis of NAD+ in yeast was thought to flow exclusively through NAMN (nicotinic acid mononucleotide).[9][10][11][12][13]
When NAD+ synthase (glutamine-hydrolysing) was deleted from yeast cells, NR permitted yeast cells to grow. Thus, these Dartmouth College investigators proceeded to clone yeast and human nicotinamide riboside kinases and demonstrate the conversion of NR to NMN by nicotinamide riboside kinases in vitro and in vivo. They also demonstrated that NR is a natural product found in cow’s milk.[14][15]
Properties
Although it is a form of vitamin B3, NR exhibits unique properties that distinguish it from the other B3 vitamins—niacin and nicotinamide. In a head-to-head experiment conducted on mice, each of these vitamins exhibited unique effects on the hepatic NAD+ metabolome with unique kinetics, and with NR as the form of B3 that produced the greatest increase in NAD+ at a single timepoint.[16]
Different biosynthetic pathways are responsible for converting the different B3 vitamins into NAD+. The enzyme nicotinamide phosphoribosyltransferase (Nampt) catalyzes the rate-limiting step of the two-step pathway converting nicotinamide to NAD+. Two nicotinamide riboside kinases (NRK1 and NRK2) convert NR to NAD+ via a pathway that does not require Nampt.[14]
Animal studies have demonstrated that these enzymes respond differently to age and stress. In a mouse model of dilated cardiomyopathy, NRK2 mRNA expression increased, while Nampt mRNA expression decreased.[17] A similar increase in NRK1 and NRK2 expression has been observed in injured central and peripheral neurons.[18][19][20][21][22]
Niacin is known for its tendency to cause an uncomfortable flushing of the skin. This flushing is triggered by the activation of the GPR109A G-protein coupled receptor. NR does not activate this receptor,[23] and has not been shown to cause flushing in humans—even at doses as high as 2,000 mg/day.[16][24][25][26]
Despite being an NAD+ precursor, nicotinamide acts as an inhibitor of the NAD+-consuming sirtuin enzymes.[10] When sirtuins consume NAD+, they create nicotinamide and O-acetyl-ADP-ribose as products of the deacetylation reaction. Consistent with high-dose nicotinamide as a sirtuin inhibitor, NR and niacin, but not nicotinamide, have been shown to increase hepatic levels of O-acetyl-ADP-ribose.[16]
Commercialization
In 2004, Dartmouth Medical School researcher Dr. Charles Brenner discovered that NR could be converted to NAD+ via the eukaryotic nicotinamide riboside kinase biosynthetic pathway[14] Dartmouth was subsequently issued patents for nutritional and therapeutic uses of NR, in 2006.[27] ChromaDex licensed these patents in July 2012, and began to develop a commercially viable, full-scale process to bring NR to market.[28]
Human Clinical Testing
There have been five published clinical trials on groups of both men and women testing for safety. One of these trials studied NR in combination with pterostilbene,[29] while the other four examined the effects of NR alone.[16][24][25][26]
The first published clinical trial established the safety and characterized the pharmacokinetics of single doses of NR.[16] Since then, doses as high as 2,000 mg/day have been administered over periods as long as 12 weeks.[25] These studies show that NR can significantly increase levels of NAD+ and some of its associated metabolites in both whole blood and peripheral blood mononuclear cells.[16][24][26]
In a 12 week clinical trial of obese insulin-resistant men using 2000 mg/day, NR appeared safe, but did not improve insulin sensitivity or whole-body glucose metabolism.[26] In a trial of NR 250 mg plus 50 mg of pterostilbene, as well as with double this dose, the combined supplement raised NAD+ levels in a trial of older adults.[29]
PATENT
WO-2019126482
Crystalline form of nicotinamide riboside chloride, useful for treating motor neuron disease or ALS, infertility, kidney damage, and liver damage or fatty liver. Elysium Health in collaboration with Mayo Clinic , is developing EH-301 (clinical, in July 2019), a combination of nicotinamide riboside chloride and pterostilbene for the treatment of amyotrophic lateral sclerosis. See WO2019108878 , claiming use of composition comprising nicotinamide riboside and pterostilbene, for treating obesity.
Nicotinamide riboside is a pyridine-nucleoside form of niacin ( i.e ., vitamin B3) that serves as a precursor to nicotinamide adenine dinucleotide (NAD+). NAD+promotes cellular metabolism, mitochondrial function, and energy production. Currently, nicotinamide riboside is made through synthetic methods or fermentation processes. Because of its significant potential to confer health benefits when used as a dietary supplement, there exists a need to develop highly efficient and scalable processes for the manufacture and purification of nicotinamide riboside.
SUMMARY OF THE INVENTION
In certain aspects, the present invention provides a crystalline form of a compound having the structure of formula (I)
Example 1. Scale-Up Synthesis and Crystallization of Nicotinamide Riboside Chloride
900 kg of nicotinamide riboside triacetate and 2133 kg of methanol were charged to a reactor and mixed, then cooled to 0 °C. 747 kg of 7M mmmonia in methanol (i.e.,“methanolic NH3”) was slowly charged to the reactor at 0 °C. The reaction mixture was passed through a polish filter, then the reaction mixture was stirred for 14 hours. A sample from the reaction mixture was taken to assess reaction progress. Upon completion of the reaction, the reaction mixture was
placed under vacuum, then warmed to 20 °C to 25 °C for 4 hours. Vacuum was applied until solids formed. Once solids were formed, the resultant slurry was filtered on a Nutsche filter dryer. Solids were washed with 1422 kg of ethanol, then 1422 kg of acetone, then 1322 kg of methyl tert butyl ether (MTBE). The resultant solids were then dried at 40 °C. Product was formed with 60% yield. The process flow diagram for this reaction is shown in FIG. 6.
Example 2. Optional Secondary Isolation
The crystalline form may optionally undergo a second isolation process according to the following steps: The solids obtained in Example 1 were dissolved in purified water at 30 °C to 40 °C. Ethanol was slowly added to the solution and mixed for 10 hours, over which time the solids began to precipitate. MTBE was then added and mixed for 2 hours. The mixture was then filtered on a Buchner funnel, and the solids were washed with ethanol, then acetone, then MTBE. Solids were dried at 40 °C.
Example 3. Spectroscopic Data.
The crystalline form made by the process described in Examples 1 and 2 has an XRD spectrum substantially as shown in FIG. 1. The instrument utilized in collecting the XRD data is a Rigaku Smart Lab X-Ray diffraction system.
Specifically, in order to collect the XRD data, The Rigaku Smart-Lab X-ray diffraction system was configured for reflection Bragg-Brentano geometry using a line source X-ray beam. The X-ray source is a Cu Long Fine Focus tube that was operated at 40 kV and 44 mA. That source provides an incident beam profile at the sample that changes from a narrow line at high angles to a broad rectangle at low angles. Beam conditioning slits are used on the line X-ray source to ensure that the maximum beam size is less than 10 mm both along the line and normal to the line. The Bragg-Brentano geometry is a para-focusing geometry controlled by passive divergence and receiving slits with the sample itself acting as the focusing component for the optics. The inherent resolution of Bragg-Brentano geometry is governed in part by the diffractometer radius and the width of the receiving slit used. Typically, the Rigaku Smart-Lab is operated to give peak widths of 0.1 °2Q or less. The axial divergence of the X-ray beam is controlled by 5.0-degree Sober slits in both the incident and diffracted beam paths.
The samples were prepared in a low background Si holder using light manual pressure to keep the sample surface flat and level with the reference surface of the sample holder. The single crystal Si low background holder has a small circular recess (10 mm diameter and about 0.2 mm depth) that held between 20 and 25 mg of the sample. The samples were analyzed from 2 to 40
°2Q using a continuous scan of 6 °20 per minute with an effective step size of 0.02 °20. The data collection procedure used to analyze these samples was not validated. The peak lists were generated using PDXL2 v.2.3.1.0. The figures were created using PlotMon VI.00.
PATENT
WO2019108878 , claiming use of composition comprising nicotinamide riboside and pterostilbene, for treating obesity.
CLIP
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0186459
CLIP
References
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- ^ Chi Y, Sauve AA (November 2013). “Nicotinamide riboside, a trace nutrient in foods, is a vitamin B3 with effects on energy metabolism and neuroprotection”. Curr Opin Clin Nutr Metab Care. 16 (6): 657–61. doi:10.1097/MCO.0b013e32836510c0. PMID 24071780.
- ^ “Nicotinamide riboside”. pubchem.ncbi.nlm.nih.gov.
- ^ “GRAS Notices, GRN No. 635”. http://www.accessdata.fda.gov. Retrieved 18 February 2019.
- ^ “Spherix/ChromaDex GRAS submission” (PDF). FDA.gov. Retrieved 18 February2019.
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- ^ Gallo; et al. (2004). “Nicotinamide clearance by pnc1 directly regulates sir2-mediated silencing and longevity”. Mol. Cell. Biol. 24 (3): 1301–1312. doi:10.1128/mcb.24.3.1301-1312.2004.
- ^ Panozzo, C.; et al. (2002). “Aerobic and anaerobic NAD+ metabolism in Saccharomyces cerevisiae”. FEBS Lett. 517 (1–3): 97–102. doi:10.1016/s0014-5793(02)02585-1. PMID 12062417.
- ^ Sandmeier, JJ; Celic, I; Boeke, JD; Smith, JS (March 2002). “Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD(+) salvage pathway”. Genetics. 160 (3): 877–89. PMC 1462005. PMID 11901108.
- ^ Jump up to:a b c Bieganowki, P. & Brenner, C. (2004). “Discoveries of Nicotinamide Riboside as a Nutrient and Conserved NRK Genes Establish a Preiss-Handler Independent Route to NAD+ in Fungi and Humans”. Cell. 117 (4): 495–502. doi:10.1016/s0092-8674(04)00416-7. PMID 15137942.
- ^ Hautkooper, R.H.; et al. (2012). “Sirtuins as regulators of metabolism and healthspan”. Nat. Rev. Mol. Cell Biol. 13 (4): 225–238. doi:10.1038/nrm3293. PMC 4872805. PMID 22395773.
- ^ Jump up to:a b c d e f Trammell, Samuel A. J.; Schmidt, Mark S.; Weidemann, Benjamin J.; Redpath, Philip; Jaksch, Frank; Dellinger, Ryan W.; Li, Zhonggang; Abel, E. Dale; Migaud, Marie E.; Brenner, Charles (10 October 2016). “Nicotinamide riboside is uniquely and orally bioavailable in mice and humans”. Nature Communications. 7 (1): 12948. doi:10.1038/ncomms12948. PMC 5062546. PMID 27721479.
- ^ Diguet, Nicolas; Trammell, Samuel A.J.; Tannous, Cynthia; Deloux, Robin; Piquereau, Jérôme; Mougenot, Nathalie; Gouge, Anne; Gressette, Mélanie; Manoury, Boris; Blanc, Jocelyne; Breton, Marie; Decaux, Jean-François; Lavery, Gareth G.; Baczkó, István; Zoll, Joffrey; Garnier, Anne; Li, Zhenlin; Brenner, Charles; Mericskay, Mathias (22 May 2018). “Nicotinamide Riboside Preserves Cardiac Function in a Mouse Model of Dilated Cardiomyopathy”. Circulation. 137 (21): 2256–2273. doi:10.1161/CIRCULATIONAHA.116.026099. PMID 29217642.
- ^ Vaur, Pauline; Brugg, Bernard; Mericskay, Mathias; Li, Zhenlin; Schmidt, Mark S.; Vivien, Denis; Orset, Cyrille; Jacotot, Etienne; Brenner, Charles; Duplus, Eric (December 2017). “Nicotinamide riboside, a form of vitamin B , protects against excitotoxicity-induced axonal degeneration”. The FASEB Journal. 31 (12): 5440–5452. doi:10.1096/fj.201700221RR. PMID 28842432.
- ^ Sasaki, Y.; Araki, T.; Milbrandt, J. (16 August 2006). “Stimulation of Nicotinamide Adenine Dinucleotide Biosynthetic Pathways Delays Axonal Degeneration after Axotomy”. Journal of Neuroscience. 26 (33): 8484–8491. doi:10.1523/JNEUROSCI.2320-06.2006. PMID 16914673.
- ^ Frederick, David W.; Loro, Emanuele; Liu, Ling; Davila, Antonio; Chellappa, Karthikeyani; Silverman, Ian M.; Quinn, William J.; Gosai, Sager J.; Tichy, Elisia D.; Davis, James G.; Mourkioti, Foteini; Gregory, Brian D.; Dellinger, Ryan W.; Redpath, Philip; Migaud, Marie E.; Nakamaru-Ogiso, Eiko; Rabinowitz, Joshua D.; Khurana, Tejvir S.; Baur, Joseph A. (August 2016). “Loss of NAD Homeostasis Leads to Progressive and Reversible Degeneration of Skeletal Muscle”. Cell Metabolism. 24 (2): 269–282. doi:10.1016/j.cmet.2016.07.005. PMC 4985182. PMID 27508874.
- ^ Cantó, Carles; Jiang, Lake Q.; Deshmukh, Atul S.; Mataki, Chikage; Coste, Agnes; Lagouge, Marie; Zierath, Juleen R.; Auwerx, Johan (March 2010). “Interdependence of AMPK and SIRT1 for Metabolic Adaptation to Fasting and Exercise in Skeletal Muscle”. Cell Metabolism. 11 (3): 213–219. doi:10.1016/j.cmet.2010.02.006. PMC 3616265. PMID 20197054.
- ^ Rappou, Elisabeth; Jukarainen, Sakari; Rinnankoski-Tuikka, Rita; Kaye, Sanna; Heinonen, Sini; Hakkarainen, Antti; Lundbom, Jesper; Lundbom, Nina; Saunavaara, Virva; Rissanen, Aila; Virtanen, Kirsi A.; Pirinen, Eija; Pietiläinen, Kirsi H. (March 2016). “Weight Loss Is Associated With Increased NAD /SIRT1 Expression But Reduced PARP Activity in White Adipose Tissue”. The Journal of Clinical Endocrinology & Metabolism. 101 (3): 1263–1273. doi:10.1210/jc.2015-3054. PMID 26760174.
- ^ Cantó, Carles; Houtkooper, Riekelt H.; Pirinen, Eija; Youn, Dou Y.; Oosterveer, Maaike H.; Cen, Yana; Fernandez-Marcos, Pablo J.; Yamamoto, Hiroyasu; Andreux, Pénélope A.; Cettour-Rose, Philippe; Gademann, Karl; Rinsch, Chris; Schoonjans, Kristina; Sauve, Anthony A.; Auwerx, Johan (June 2012). “The NAD+ Precursor Nicotinamide Riboside Enhances Oxidative Metabolism and Protects against High-Fat Diet-Induced Obesity”. Cell Metabolism. 15 (6): 838–847. doi:10.1016/j.cmet.2012.04.022. PMC 3616313. PMID 22682224.
- ^ Jump up to:a b c Airhart, Sophia E.; Shireman, Laura M.; Risler, Linda J.; Anderson, Gail D.; Nagana Gowda, G. A.; Raftery, Daniel; Tian, Rong; Shen, Danny D.; O’Brien, Kevin D.; Sinclair, David A. (6 December 2017). “An open-label, non-randomized study of the pharmacokinetics of the nutritional supplement nicotinamide riboside (NR) and its effects on blood NAD+ levels in healthy volunteers”. PLOS ONE. 12 (12): e0186459. doi:10.1371/journal.pone.0186459. PMC 5718430. PMID 29211728.
- ^ Jump up to:a b c Dollerup, Ole L; Christensen, Britt; Svart, Mads; Schmidt, Mark S; Sulek, Karolina; Ringgaard, Steffen; Stødkilde-Jørgensen, Hans; Møller, Niels; Brenner, Charles; Treebak, Jonas T; Jessen, Niels (August 2018). “A randomized placebo-controlled clinical trial of nicotinamide riboside in obese men: safety, insulin-sensitivity, and lipid-mobilizing effects”. The American Journal of Clinical Nutrition. 108 (2): 343–353. doi:10.1093/ajcn/nqy132. PMID 29992272.
- ^ Jump up to:a b c d Martens, Christopher R.; Denman, Blair A.; Mazzo, Melissa R.; Armstrong, Michael L.; Reisdorph, Nichole; McQueen, Matthew B.; Chonchol, Michel; Seals, Douglas R. (29 March 2018). “Chronic nicotinamide riboside supplementation is well-tolerated and elevates NAD+ in healthy middle-aged and older adults”. Nature Communications. 9 (1): 1286. doi:10.1038/s41467-018-03421-7. PMC 5876407. PMID 29599478.
- ^ Brenner, Charles (20 April 2006). “Nicotinamide riboside kinase compositions and methods for using the same”. Google Patents. Dartmouth College. Retrieved 19 February2019.
- ^ “ChromaDex Licenses Exclusive Patent Rights for Nicotinamide Riboside (NR) Vitamin Technologies”. 2012-07-16. Retrieved 15 February 2019.
- ^ Jump up to:a b Dellinger, Ryan W.; Santos, Santiago Roel; Morris, Mark; Evans, Mal; Alminana, Dan; Guarente, Leonard; Marcotulli, Eric (24 November 2017). “Repeat dose NRPT (nicotinamide riboside and pterostilbene) increases NAD+ levels in humans safely and sustainably: a randomized, double-blind, placebo-controlled study”. NPJ Aging and Mechanisms of Disease. 3 (1): 17. doi:10.1038/s41514-017-0016-9. PMC 5701244. PMID 29184669.
Further reading
- “Press Release: NIH researchers find potential target for reducing obesity-related inflammation”. National Institutes of Health (NIH). 16 November 2015.
- Stipp, David (March 11, 2015). “Guest Blog: Beyond Resveratrol: The Anti-Aging NAD Fad”. Scientific American Blog Network.
- Zhang, H; Ryu, D; Wu, Y; Gariani, K; Wang, X; Luan, P; D’Amico, D; Ropelle, ER; Lutolf, MP; Aebersold, R; Schoonjans, K; Menzies, KJ; Auwerx, J (17 June 2016). “NAD⁺ repletion improves mitochondrial and stem cell function and enhances life span in mice”. Science. 352 (6292): 1436–43. doi:10.1126/science.aaf2693. PMID 27127236.
- Dolopikou CF, Kourtzidis IA, Margaritelis NV, Vrabas IS, Koidou I, Kyparos A, Theodorou AA, Paschalis V, Nikolaidis MG. (2019 Feb 6). Acute nicotinamide riboside supplementation improves redox homeostasis and exercise performance in old individuals: a double-blind cross-over study. doi:10.1007/s00394-019-01919-4.
ADDITIONAL INFORMATION
High dose nicotinic acid is used as an agent that elevates high-density lipoprotein cholesterol, lowers low-density lipoprotein cholesterol and lower free fatty acids through a mechanism that is not completely understood. It was suggested that nicotinamide riboside might possess such an activity by elevating NAD in the cells responsible for reverse cholesterol transport. The discovery that the Wallerian degeneration slow gene encodes a protein fusion with NMN adenylyltransferase 1 indicated that increased NAD+ precursor supplementation might oppose neurodegenerative processes.
ChromaDex acquired intellectual property on uses and synthesis of NR from Dartmouth College, Cornell University, and Washington University and began distributing NR as Niagen in 2013. In November 2015 ChromaDex received New Dietary Ingredient (NDI) status for Niagen from the U.S. Food and Drug Administration (FDA) and the FDA issued a generally recognized as safe (GRAS) No Objection Letter for Nicotinamide Riboside Chloride (NR) on August 3, 2016.
REFERENCES
1: Chi Y, Sauve AA. Nicotinamide riboside, a trace nutrient in foods, is a vitamin B3 with effects on energy metabolism and neuroprotection. Curr Opin Clin Nutr Metab Care. 2013 Nov;16(6):657-61. doi: 10.1097/MCO.0b013e32836510c0. Review. PubMed PMID: 24071780.
2: Bogan KL, Brenner C. Nicotinic acid, nicotinamide, and nicotinamide riboside: a molecular evaluation of NAD+ precursor vitamins in human nutrition. Annu Rev Nutr. 2008;28:115-30. doi: 10.1146/annurev.nutr.28.061807.155443. Review. PubMed PMID: 18429699.
3: Ghanta S, Grossmann RE, Brenner C. Mitochondrial protein acetylation as a cell-intrinsic, evolutionary driver of fat storage: chemical and metabolic logic of acetyl-lysine modifications. Crit Rev Biochem Mol Biol. 2013 Nov-Dec;48(6):561-74. doi: 10.3109/10409238.2013.838204. Review. PubMed PMID: 24050258; PubMed Central PMCID: PMC4113336.
4: Yang Y, Sauve AA. NAD(+) metabolism: Bioenergetics, signaling and manipulation for therapy. Biochim Biophys Acta. 2016 Dec;1864(12):1787-1800. doi: 10.1016/j.bbapap.2016.06.014. Review. PubMed PMID: 27374990.
5: Sauve AA. NAD+ and vitamin B3: from metabolism to therapies. J Pharmacol Exp Ther. 2008 Mar;324(3):883-93. doi: 10.1124/jpet.107.120758. Review. PubMed PMID: 18165311.
6: Kato M, Lin SJ. Regulation of NAD+ metabolism, signaling and compartmentalization in the yeast Saccharomyces cerevisiae. DNA Repair (Amst). 2014 Nov;23:49-58. doi: 10.1016/j.dnarep.2014.07.009. Review. PubMed PMID: 25096760; PubMed Central PMCID: PMC4254062.
7: Gerlach G, Reidl J. NAD+ utilization in Pasteurellaceae: simplification of a complex pathway. J Bacteriol. 2006 Oct;188(19):6719-27. Review. PubMed PMID: 16980474; PubMed Central PMCID: PMC1595515.
8: Srivastava S. Emerging therapeutic roles for NAD(+) metabolism in mitochondrial and age-related disorders. Clin Transl Med. 2016 Dec;5(1):25. doi: 10.1186/s40169-016-0104-7. Review. PubMed PMID: 27465020; PubMed Central PMCID: PMC4963347.
9: Handschin C. Caloric restriction and exercise “mimetics”: Ready for prime time? Pharmacol Res. 2016 Jan;103:158-66. doi: 10.1016/j.phrs.2015.11.009. Review. PubMed PMID: 26658171; PubMed Central PMCID: PMC4970791.
10: Ruggieri S, Orsomando G, Sorci L, Raffaelli N. Regulation of NAD biosynthetic enzymes modulates NAD-sensing processes to shape mammalian cell physiology under varying biological cues. Biochim Biophys Acta. 2015 Sep;1854(9):1138-49. doi: 10.1016/j.bbapap.2015.02.021. Review. PubMed PMID: 25770681.
11: Imai S, Guarente L. NAD+ and sirtuins in aging and disease. Trends Cell Biol. 2014 Aug;24(8):464-71. doi: 10.1016/j.tcb.2014.04.002. Review. PubMed PMID: 24786309; PubMed Central PMCID: PMC4112140.
12: Jaehme M, Slotboom DJ. Structure, function, evolution, and application of bacterial Pnu-type vitamin transporters. Biol Chem. 2015 Sep;396(9-10):955-66. doi: 10.1515/hsz-2015-0113. Review. PubMed PMID: 26352203.
13: Magni G, Di Stefano M, Orsomando G, Raffaelli N, Ruggieri S. NAD(P) biosynthesis enzymes as potential targets for selective drug design. Curr Med Chem. 2009;16(11):1372-90. Review. PubMed PMID: 19355893.
14: Mendelsohn AR, Larrick JW. Partial reversal of skeletal muscle aging by restoration of normal NAD⁺ levels. Rejuvenation Res. 2014 Feb;17(1):62-9. doi: 10.1089/rej.2014.1546. Review. PubMed PMID: 24410488.
15: Penberthy WT. Pharmacological targeting of IDO-mediated tolerance for treating autoimmune disease. Curr Drug Metab. 2007 Apr;8(3):245-66. Review. PubMed PMID: 17430113.
16: Gazzaniga F, Stebbins R, Chang SZ, McPeek MA, Brenner C. Microbial NAD metabolism: lessons from comparative genomics. Microbiol Mol Biol Rev. 2009 Sep;73(3):529-41, Table of Contents. doi: 10.1128/MMBR.00042-08. Review. PubMed PMID: 19721089; PubMed Central PMCID: PMC2738131.
17: Magni G, Amici A, Emanuelli M, Orsomando G, Raffaelli N, Ruggieri S. Enzymology of NAD+ homeostasis in man. Cell Mol Life Sci. 2004 Jan;61(1):19-34. Review. PubMed PMID: 14704851.
18: Magni G, Orsomando G, Raffelli N, Ruggieri S. Enzymology of mammalian NAD metabolism in health and disease. Front Biosci. 2008 May 1;13:6135-54. Review. PubMed PMID: 18508649.
19: Belenky P, Bogan KL, Brenner C. NAD+ metabolism in health and disease. Trends Biochem Sci. 2007 Jan;32(1):12-9. Review. Erratum in: Trends Biochem Sci. 2008 Jan;33(1):1. PubMed PMID: 17161604.
20: Niven DF, O’Reilly T. Significance of V-factor dependency in the taxonomy of Haemophilus species and related organisms. Int J Syst Bacteriol. 1990 Jan;40(1):1-4. Review. PubMed PMID: 2145965.
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| Names | |
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| Other names
1-(β-D-Ribofuranosyl)nicotinamide; N-Ribosylnicotinamide
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| Identifiers | |
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3D model (JSmol)
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| ChEBI | |
| ChemSpider | |
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PubChem CID
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| Properties | |
| C11H15N2O5+ | |
| Molar mass | 255.25 g/mol |
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Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
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///////////// EH-301, EH 301, EH301, Nicotinamide riboside, SRT647, SRT-647, SRT 647, Nicotinamide Riboside Triflate, α/β mixture
C1=CC(=C[N+](=C1)C2C(C(C(O2)CO)O)O)C(=O)N.[Cl-]
New patent, Opicapone, WO 2019123066, Unichem
New patent, Opicapone, WO 2019123066, Unichem
WO-2019123066
https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2019123066&tab=PCTDESCRIPTION
Process for the preparation of opicapone and its intermediates, useful for treating parkinson’s disease. Bial-Portela has developed and launched opicapone, for treating Parkinson’s disease.
Opicapone is a selective and reversible catechol-O-methyltransferase (COMT) inhibitor, use as adjunctive therapy for parkinson’s disease. Opicapone was approved by European Medicine Agency (EMA) on June 24, 2016 and it is developed and marketed as ONGENTYS® by Bial-Portela in Europe. Opicapone is chemically described as 2,5-dichloro-3-(5-(3,4-dihydroxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl)-4,6-dimethylpyridine-l-oxide and depicted below as compound of formula (I).
Opicapone and a process for preparation of it is disclosed in US 8,168,793. The process disclosescondensation of 3, 4-dibenzyloxy-5-nitrobenzoic acid with (Z)-2, 5-dichloro-N’-hydroxy-4, 6-dimethylnicotinimidamide in presence of N, N’-Carbonyl diimidazole in N, N’-dimethylformamide. The crude condensation intermediate was subjected to tetrabutylammonium fluoride (TBAF) mediated cyclization in tetrahydrofuran to give l,2,4-oxadiazole derivative, purifying it by precipitating in 1:1 mixture of dichloromethane: diethyl ether and recrystallized it in isopropyl alcohol. Oxidation of l,2,4-oxadiazole compound is carried out using 10 fold excess of urea hydrogen peroxide complex and trifluoroacetic anhydride in dichloromethane and was purified by column chromatography. Obtained N-oxide compound was converted into opicaponecompound of formula (I) by deprotection O-benzoyl groups by exposure it to boron tribromide (BBr3) in dichloromethane at -78°C to room temperature. Final product was purified in mixture of toluene and ethanol. Above synthetic stepsare outline in scheme 1.
c eme
This process has several drawbacks like cyclization reaction involve use of TBAF and THF. Use of expensive TBAF, leads to high cost in the production and therefore uneconomical for industrial production, whereas use of THF during this reaction has limitation due to peroxide contents. . Similarly diethyl ether is a potential fire hazard and can form peroxides rapidly and thus should be avoided in commercial scale production. Above cyclization is also carried out in presence of DMF and CDI at l20°C.
Similar approach was reported in W02008094053 which describes preparation of opicapone by one pot cyclization of 3,4-dibenzyloxy-5-nitrobenzoic acid with (Z)-2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide using N,N’-Carbonyl diimidazole in N,N’-dimethylformamide followed by heating the reaction mixture at l35°C for 5 hours to obtain l,2,4-oxadiazole derivative. This oxadiazole derivative was purified by recrystallization from isopropyl alcohol. Further oxidation using urea hydrogen peroxide complex followed by o-debenzylation using boron tribromide (BBr3) was achieved to obtain Opicapone.
This process also suffers from drawback like use of elevated temperature (l35°C) and use of expensive BBr3.
US 9,126,988 also disclose process for the preparation of opicapone, whichinvolves several chemical steps: 1) nitrating vanillic acid in presence of nitric acid in acetic acidfollowed by recrystallization with acetic acid to get nitro compound with yield 40-46%; 2)which converted into acid chloride compoundby treating it with thionyl chloride in presence of catalytic amount of N, N-dimethylformamide in dichloromethane or l,4-dioxane; 3) condensing acid chloride compound with (Z)-2, 5-dichloro-N’-hydroxy-4, 6-dimethylnicotinimidamide in presence of excess amount of pyridine in N,N-dimethyl acetamide/ tetrahydrofuran/ dichloromethane or l,4-dioxane at 5-10 °C and then heating the reaction mixture at H0-l l5°C for 5-6 hours to get 1,2,4-oxadiazole compound; 4) which was oxidized using urea hydrogen peroxide complex and trifluoroacetic anhydride in dichloromethane to get N-oxide product which was purified by repeated recrystallization (2 or more times) using mixture of formic acid and toluene to get pure product with 59% yield; 5) O-methyl group was deprotected using aluminium chloride and pyridine in N-Methyl pyrrolidone at 60 C to obtain opicapone. After completion of reaction, the crude product was isolated by quenching the reaction mixture in mixture cone. HC1: water followed by filtration, washing with water: isopropyl alcohol and recrystallization from ethanol. Final purification was done in mixture of formic acid and isopropyl alcohol. Above synthetic steps are outline in scheme 2.
Scheme 2
As described above, cited literature processes suffers from some drawbacks like elevated reaction temperature and longer duration, use of excess amount of pyridine for cyclization reaction which is difficult to handle on large scale preparation. Another drawback of reported procedure is unsafe workup procedures for isolation of N-oxide as residual peroxides were not quenched by any peroxide quenching reagent. Also repeated crystallizations (more than two) are required for purification of N-oxide derivative to remove unreacted starting material which is tedious and time consuming process. Also for its purification mixture of solvent i.e. formic acid and toluene are used which hamper its recovery and is not cost effective process.
US 9,126,988 also disclosed process for the preparation of 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide compound of formula (IV), in which 2,5-dichloro-4,6-dimethylnicotinonitrile compound of formula (VIII) was reacted with hydroxyl amine solution in the presence of catalytic amount of 1,10-phenanthroline in methanokwater at 70-80°C for 6 hrs. After completion reaction mixture was cooled, filtered and dried to get 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (88%).
Bioorganic & Medicinal Chemistry 13 (2005) 5740-5749, Karl Bailey et.al. disclosed process for preparation of 3,4-dimethoxy-5-nitro benzoic acid compound of formula (Ilia). In which a solution of Cr03, concentrated H2S04 and water was added to solution of 3,4-dimethoxy-5-nitro benzaldehyde in acetone and water. The obtained solution was stirred for 24 hrs and then isopropanol was added to eliminate any unreacted Cr(VI) species to obtained crude green sludge, which was extracted into ethyl acetate and washed with 1M HC1 to remove remaining Cr(III) species. Obtained product is then recrystallized from water and ethanol to yield 69 % of 3,4-dimethoxy-5-nitro benzoic acid compound of formula (Ilia).
US 5,358, 948 also disclosed process for preparation of 3,4-dimethoxy-5-nitro benzoic acid compound of formula (Ilia). In which a solution of potassium permanganate was added to a solution of 3,4-dimethoxy-5-nitro benzaldehyde in acetone. The mixture was then stirred at 20°C for 18 hrs togives 3,4-dimethoxy-5-nitro benzoic acid compound of formula (Ilia) with 72% yield.
Disadvantage of the above cited literature (Karl Bailey et.al and US’ 948) processes are harsh, acidic condition and involve expensive reagents. The process is both uneconomical and time consuming, (18-24 hrs) hence not suitable for commercial production.
Oxidation of aldehydes to the corresponding carboxylic acids, on the other hand, are commonly carried out using KMn04 in acidic or basic media, or K2Cr207 in acidic medium or chromic acid. These heavy metal-based reagents are hazardous and the protocols produce metal wastes that require special handling owing to their toxicities.
It is therefore, desirable to provide efficient, robust, alternative simple process, cost effective process which is used on a large scale and allows product to be easily workup, purified and isolate without the disadvantages mentioned above.
Example 1: Preparation of 3, 4-dimethoxy-5-nitro benzoic acid (Ilia).
To a cooled solution of 3,4-dimethoxy-5-nitro benzaldehyde (lOOg, 0.474 mole) in DMF (500 ml) was added Oxone (294.1 g, 0.478 mole) lot wise at 5-10 °C. Reaction mixture was stirred for 30 minutes at same temperature, allowed to warm to room temperature and stirred for 2-3 hours. After completion, the reaction mixture was diluted with 1500 ml of water and filtered. The solid was washed with water until all peroxides removed and drying at 50°C under vacuum afforded 3,4-dimethoxy-5-nitro benzoic acid of formula (Ilia) (l02g, 95%).
Example 2: Preparation of 2 ,5-dichloro-N’ {[(3,4-dimethoxy-5-nitrophenyl) carbonyl]oxy}-4, 6-dimethylpyridine-3-carboximidamide (Va)
To a solution of 3,4-dimethoxy-5-nitro benzoic acid of formula (Ilia) (5 g, 0.022 mole)in 60 ml of acetonitrile was added N,N’-Carbonyldiimidazole (4.28g, 0.026 mole) in portions and the reaction mixture was stirred at room temperature for 1.5 hours. Then was added 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (5.4g, 0.023 mole) and stirring was continued for 3 hours. After completion, the reaction mixture was diluted with 240 ml of water and 300 ml of dichloromethane. Organic layer was separated and washed with water (200 ml x 3), concentrated under reduced pressure to obtain 2,5-dichloro-N'{ [(3,4-dimethoxy-5-nitrophenyl)carbonyl]oxy}-4,6-dimethylpyridine-3-carboximidamide of formula (Va) (8.67g, 88.9%).
Example 3: Preparation of 2, 5-dichloro-3-[5-(3, 4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine (Via)
To a solution of 2,5-dichloro-N'{ [(3,4-dimethoxy-5-nitrophenyl)carbonyl]oxy}- 4,6-dimethylpyridine-3-carboximidamide of formula (Va) (0.5g, 0.0011 mole)in 10 ml of dichloromethane was added isopropyl alcohol (1 ml) followed by KOH (0.075g, 0.001 lmole) dissolved in 0.1 ml of water. After stirring for 1 hour at room temperature the reaction mixture was diluted with 30 ml of dichloromethane and washed with water (lOml x 2). The reaction mixture was concentrated under reduced pressure to obtain 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine of formula (Via) (0.4 g, 83%).
Example 4: Preparation of 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine (Via) (One pot cyclization procedure)
To a stirred solution of 3,4-dimethoxy-5-nitro benzoic acid formula (Ilia) (lOOg, 0.44 mol)in 1000 ml of dichloromethane was added N,N’-Carbonyldiimidazole (86g, 0.53 mole) in portions and the reaction mixture was stirred at room temperature for 1.5 hours. Then was added 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (l08g, 0.46 mole) and stirring was continued for 3 hours. Isopropyl alcohol (200 ml) and KOH (30g, 0.53 mole) dissolved in 30 ml of water was then added to the reaction mixture. After stirring for 1 hour at room temperature the organic layer was washed with water (1000 ml x 2). Solvent was distilled out at atmospheric pressure, added 1000 ml of isopropyl alcohol and suspension was stirred at 55-60°C for 2 hours. The reaction mixture was allowed to cool to room temperature, stirred for 2 hours and filtered. The solid was washed with isopropyl alcohol (100 ml x 2) and dried at 50-60°C under vacuum to obtain 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine of formula (Via) (l60g, 85%).
Example 5: Preparation of 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine (Via) (cyclization procedure using thionyl chloride)
To a stirred solution of 3,4-dimethoxy-5-nitro benzoic acid of formula (Ilia) (lOOg, 0.44 mol) in 500 ml of dichloromethane was added 0.4 ml of N,N-dimethyl formamide followed bythionyl chloride (82g, 0.69 mole) drop wise at room temperature and the reaction mixture was heated at 40°C for 4 hours. After completion, dichloromethane and excess of thionyl chloride was distilled out under reduced pressure at 40°C. The obtained residue was dissolved in 500 ml of dichloromethane and was added to pre-cooled mixture of 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (l03g, 0.44 mole) and triethyl amine (73 ml, 0.53 mole) in 500 ml of dichloromethane at 5°C. After addition, the reaction mixture was allowed to warm to 25-30°C and stirred for 2 hours. Then was added isopropyl alcohol (200 ml) followed by KOH (62g, 1.1 mole) dissolved in 62 ml of water and stirring was continued for 2 hours at room temperature. The reaction mixture was washed with 1000 ml of water, 1N aqueous HC1 solution (500ml x 2) followed by 500 ml of 5% aqueous sodium bicarbonate solution. Solvent was distilled out at atmospheric pressure at 40°C. To the residue was added 1200 ml of methanol and the suspension was stirred at 55-60°C for 2 hours. The reaction mixture was allowed to cool to room temperature, maintained for 2 hours and filtered. The solid product was washed with methanol (100 ml x 2) and dried at 50°C under vacuum to obtain 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)- 1, 2, 4-oxadiazol-3-yl]-4, 6-dimethyl pyridine of formula (Via) (l65g, 88%).
Example 6: Preparation of 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine-l-oxide (Vila)
To a cooled solution of 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine of formula (Via) (25g, 0.0588 mole) in 300 ml of dichloromethane was added urea hydrogen peroxide complex (l8.26g, 0.194 mole) in portions followed by trifluoroacetic anhydride (37g, 0.176 mole) maintaining temperature below l0°C. After stirring at 5-l0°C for 1 hour, the reaction mixture was allowed to warm to room temperature and stirred for 5 hours. The reaction mixture was washed with water (300 ml x 2), 300ml of 5% aqueous sodium sulphite solution to quench residual peroxides and finally with 300 ml of water. Dichloromethane layer was distilled out at atmospheric pressure. The obtained solid was suspended in 250 ml of ethyl acetate and 12.5 ml of cone. HC1 was added at room temperature. The resulting suspension was then stirred at 65-70°C for 1 hour and allowed to cool to room temperature. After stirring for 2 hours, the reaction mixture was filtered, solid was washed with ethyl acetate (50 ml x 2) followed by water (50 x 3) and dried at 50°C under vacuum to obtain (5-(3,4-bis(methoxy)-5-nitrophenyl)-l,2,4-oxadiazol-3-yl)-2,5-dichloro-4,6-dimethylpyridine 1 -oxide of formula (Vila) (18g, 69%).
Example 7: Preparation of 5-[3-(2,5-Dichloro-4,6-dimethyl-l-oxido-3-pyridinyl)-l,2,4-oxadiazol-5-yl]-3-nitro-l,2-benzenediol (Opicapone, I)
To a cooled solution of 2,5-dichloro-3-[5-(3,4-dimethoxy-5-nitrophenyl)-l,2,4-oxadiazol-3-yl]-4,6-dimethylpyridine-l-oxide of formula (Vila) (25g, 0.056 mole) in 200 ml of N,N-Dimethylformamide was added AlCl3 (l l.34g, 0.085 mol) at 5-l0°C in portions. The reaction mixture was then heated at 85 °C for 6 hours. After completion, the reaction mixture was cooled to room temperature and poured onto cold mixture of cone. HC1 (200 ml) and water (400 ml). The reaction mixture was filtered, solid washed with water (100 ml X 3) followed by methanol (50 ml x2) and dried at 50°C under vacuum to obtain 5-[3-(2,5-Dichloro-4, 6-dimethyl- l-oxido-3-pyridinyl)- 1,2, 4-oxadiazol-5-yl]-3-nitro- 1,2-benzenediol of formula (I) (22 g, 94%).
Example 8: Preparation of 2, 5-dichloro-N’-hydroxy-4, 6-dimethyl nicotinimidamide of formula (IV)
To a suspension of 2,5-dichloro-4,6-dimethylnicotinonitrile of formula (VIII) (lOOg, 0.497 mole) in l,4-dioxane (400 ml) and water (900 ml) was added 50% aqueous solution of hydroxyl amine (l30g) and N-methyl morpholine (50.2g, 0.497) at room temperature. The reaction mixture was then stirred at 70-80°C for 10 hours. After completion, water (1100 ml) was added to the reaction mixture at 70-80°C and allowed to cool to room temperature. After stirring for 2 hours the reaction mixture was filtered, solid was washed with water (200ml x 3) and dried at 50°C under vacuum to obtain 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (68 g, 58%).
Example 9:Preparation of 2,5-dichloro-N’-hydroxy-4, 6-dimethylnicotinimidamide of formula (IV)
To a suspension of 2,5-dichloro-4,6-dimethylnicotinonitrile of formula (VIII) (lOOg, 0.497 mole) in methanol (600 ml) and water (800 ml) was added 50% aqueous solution of hydroxyl amine (l30g) and 2-methylpyrazine (7.02g, 0.0746) at room temperature. The reaction mixture was then stirred at 70-80°C for 6-8 hours. After completion, water (800 ml) was added to the reaction mixture at 70-80°C and allowed to cool to room temperature. After stirring for 2 hours the reaction mixture was filtered, solid was washed with water (200ml x 3) and dried at 50°C under vacuum to obtain 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (82 g, 70%).
Example 10: Preparation of 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV)
To a solution of hydroxylamine hydrochloride (86.4g, 1.243 mole) in 400 ml of water was added LiOH.H20 (52.7g, 1.25 mole) at room temperature and heated at 50°C for 30 minutes. To the reaction mixture was added 300 ml of methanol, 2-methylpyrazine (3.5 lg, 0.037 mole) and 2,5-dichloro-4,6-dimethylnicotinonitrile of formula (VIII) (50g, 0.248 mole) at 50°C. The reaction mixture was then stirred at 70-80°C for 6 hours. After completion, water (500 ml) was added to the reaction mixture at 70-80°C and allowed to cool to room temperature. After stirring for 2 hours the reaction mixture was filtered, solid was washed with water (lOOml x 3) and dried at 50°C under vacuum to obtain 2,5-dichloro-N’-hydroxy-4,6-dimethylnicotinimidamide of formula (IV) (37.6 g, 64%).
Example 11: Purification of 5-[3-(2,5-Dichloro-4,6-dimethyl-l-oxido-3-pyridinyl)-l,2,4-oxadiazol-5-yl]-3-nitro-l,2-benzenediol (Opicapone, I)
The crude 5-[3-(2,5-Dichloro-4, 6-dimethyl- l-oxido-3-pyridinyl)- 1,2, 4-oxadiazol-5-yl]-3-nitro-l,2-benzenediol of formula (I)(25.0g) was suspended in 250 ml of N,N-dimethylformamide and reaction mixture was heated at 60-65°C to obtain clear solution. Then was added 500 ml of methanol and reaction mixture was cooled to room temperature. After stirring for 2-3 hours, the reaction mixture was filtered, solid was washed with methanol and dried at 50°C under vacuum to obtain 5-[3-(2, 5-Dichloro-4, 6-dimethyl- l-oxido-3-pyridinyl)- 1,2, 4-oxadiazol-5-yl]-3-nitro-l,2-benzenediol of formula (I) (22.0 g, 88%).
/////////////New patent, Opicapone, WO 2019123066, Unichem, WO2019123066
Quinupramine, キヌプラミン
Quinupramine
キヌプラミン
- 5-(1-azabicyclo[2.2.2]oct-3-yl)-10,11-dihydro-5H-dibenz[b,f]azepine
- Formula:C21H24N2
- MW:304.44 g/mol
- CAS:31721-17-2
Quinupramine (brand names Kevopril, Kinupril, Adeprim, Quinuprine) is a tricyclic antidepressant (TCA) used in Europe for the treatment of depression.[1][2]
Pharmacologically, quinupramine acts in vitro as a strong muscarinic acetylcholine receptor antagonist (anticholinergic) and H1 receptorantagonist (antihistamine), moderate 5-HT2 receptor antagonist, and weak serotonin and norepinephrine reuptake inhibitor.[3] It has negligible affinity for the α1-adrenergic, α2-adrenergic, β-adrenergic, or D2 receptor.[3]
Clinically, quinupramine is reported to be stimulating similarly to imipramine, desipramine, and demexiptiline.[4] It can be inferred that its in vivo metabolites may have stronger effects on the reuptake of norepinephrine and/or serotonin than quinupramine itself
SYN
References
- ^ Swiss Pharmaceutical Society (2000). Index Nominum 2000: International Drug Directory (Book with CD-ROM). Boca Raton: Medpharm Scientific Publishers. p. 908. ISBN 3-88763-075-0.
- ^ José Miguel Vela; Helmut Buschmann; Jörg Holenz; Antonio Párraga; Antoni Torrens (2007). Antidepressants, Antipsychotics, Anxiolytics: From Chemistry and Pharmacology to Clinical Application. Weinheim: Wiley-VCH. p. 248. ISBN 978-3-527-31058-6.
- ^ Jump up to:a b Sakamoto H, Yokoyama N, Kohno S, Ohata K (December 1984). “Receptor binding profile of quinupramine, a new tricyclic antidepressant”. Japanese Journal of Pharmacology. 36 (4): 455–60. doi:10.1254/jjp.36.455. PMID 6098759.
- ^ Kent, Angela; M. Billiard (2003). Sleep: physiology, investigations, and medicine. New York: Kluwer Academic/Plenum. p. 233. ISBN 0-306-47406-9.
-
- DOS 2 030 492 (Sogeras; appl. 20.6.1970; GB-prior. 20.6.1969).
- GB 1 252 320 (Sogeras; valid from 29.5.1970; prior. 20.6.1969).
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| Clinical data | |
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| Routes of administration |
Oral |
| ATC code | |
| Legal status | |
| Legal status |
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| Pharmacokinetic data | |
| Elimination half-life | 33 hours |
| Identifiers | |
| CAS Number | |
| PubChem CID | |
| ChemSpider | |
| UNII | |
| KEGG | |
| ECHA InfoCard | 100.046.149 |
| Chemical and physical data | |
| Formula | C21H24N2 |
| Molar mass | 304.43 g/mol g·mol−1 |
//////////////Quinupramine, キヌプラミン
Quizartinib dihydrochloride, キザルチニブ塩酸塩
Quizartinib dihydrochloride
キザルチニブ塩酸塩
| Formula |
C29H32N6O4S. 2HCl
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|---|---|
| CAS |
1132827-21-4
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| Mol weight |
633.5891
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JAPAN, PMDA APPROVED,. 2019/6/18, Vanflyta, To use as part of a treatment regimen for newly diagnosed acute myeloid leukemia that meets certain criteria
Drug Trials Snapshot
fda 2023, 7/20/2023
Quizartinib (AC220) is a small molecule receptor tyrosine kinase inhibitor, originated Ambit Biosciences, and acquired by Daiichi Sankyo, that is currently under development for the treatment of acute myeloid leukaemia. Its molecular target is FLT3, also known as CD135 which is a proto-oncogene.[1]
Flt3 mutations are among the most common mutations in acute myeloid leukaemia due to internal tandem duplication of Flt3. The presence of this mutation is a marker of adverse outcome.
Mechanism
Specifically, Quizartinib selectively inhibits class III receptor tyrosine kinases, including FMS-related tyrosine kinase 3 (FLT3/STK1), colony-stimulating factor 1 receptor (CSF1R/FMS), stem cell factor receptor (SCFR/KIT), and platelet derived growth factor receptors (PDGFRs).
Mutations cause constitutive action of Flt3 resulting in inhibition of ligand-independent leukemic cell proliferation and apoptosis.
Clinical trials
It reported good results in 2012 from a phase II clinical trial for refractory AML – particularly in patients who went on to have a stem cell transplant.[2]
As of 2017 it has completed 5 clinical trials and another 7 are active.[3]
SYN
References
- ^ Chao, Qi; Sprankle, Kelly G.; Grotzfeld, Robert M.; Lai, Andiliy G.; Carter, Todd A.; Velasco, Anne Marie; Gunawardane, Ruwanthi N.; Cramer, Merryl D.; Gardner, Michael F.; James, Joyce; Zarrinkar, Patrick P.; Patel, Hitesh K.; Bhagwat, Shripad S. (2009). “Identification of N-(5-tert-Butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,1-b][1,3]benzothiazol-2-yl]phenyl}urea Dihydrochloride (AC220), a Uniquely Potent, Selective, and Efficacious FMS-Like Tyrosine Kinase-3 (FLT3) Inhibitor”. Journal of Medicinal Chemistry. 52 (23): 7808–7816. doi:10.1021/jm9007533.
- ^ Drug Tames Refractory AML. ASH Dec 2012
- ^ Quizartinib studies
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|
| Names | |
|---|---|
| IUPAC name
1-(5-(tert-Butyl)isoxazol-3-yl)-3-(4-(7-(2-morpholinoethoxy)benzo[d]imidazo[2,1-b]thiazol-2-yl)phenyl)urea
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| Other names
AC220
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| Identifiers | |
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3D model (JSmol)
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| ChEBI | |
| ChEMBL | |
| ChemSpider | |
| KEGG | |
| UNII | |
|
CompTox Dashboard (EPA)
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| Properties | |
| C29H32N6O4S | |
| Molar mass | 560.67 g·mol−1 |
|
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
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/////////////Quizartinib dihydrochloride, キザルチニブ塩酸塩, JAPAN 2019
OLD POST
N-(5-tert-butyl-isoxazol-3-yl)-N’-{ 4- [7-(2-morpholin-4-yl-ethoxy)imidazo [2, 1 -b] [ 1 ,3 ]benzothiazol-2-yl]phenyl } urea
| CAS | 950769-58-1 (free base) 1132827-21-4 (2HCl) |
| Formula | C29H32N6O4S |
| MW | 560.7 |
| Synonim | AC220, AC-010220 ASP-2689 |
Quizartinib
Ambit Biosciences
Ambit Biosciences (NASDAQ:AMBI) is a biotech company that focuses on treatments that inhibit kinases, which are drivers for diseases such as cancer. Three drugs are in development, with the lead one being quizartinib — a Phase 2B trial treatment for acute myeloid leukemia. However, AMBI’s collaboration agreement with Astellas Pharma is set to expire in September, and if it is not replaced, it could mean a delay in Phase 3 trials for quizartinib. Keep in mind that AMBI generated $23.8 million in collaboration revenues last year.
Quizartinib (AC220) is a small molecule receptor tyrosine kinase inhibitor that is currently under development by Ambit Biosciencesfor the treatment of acute myeloid leukaemia. Its molecular target is FLT3, also known as CD135 which is a proto-oncogene.[1]
AC-220 is an angiogenesis inhibitor that antagonizes several proteins involved in vascularization. It was engineered by Ambit Biosciences using KinomeScan technology to potently target FLT3, KIT, CSF1R/FMS, RET and PDGFR kinases. Ambit is developing oral AC-220 in phase III clinical studies for the treatment of relapsed/refractory acute myeloid leukemia (AML) patients with the FMS-like tyrosine kinase-3 (FLT3)-ITD mutation. Early clinical trials are also ongoing for the treatment of advanced solid tumors, for the treatment of refractory or relapsed myelodysplasia, in combination with induction and consolidation chemotherapy for previously-untreated de novo acute myeloid leukemia, and as a maintenance therapy of AML following hematopoietic stem cell transplantation (HSCT). In 2009, orphan drug designation was received both in the U.S. and in the EU for the treatment of AML. In 2009, Ambit Biosciences and Astellas Pharma have entered into a worldwide agreement to jointly develop and commercialize the drug candidate for the treatment of cancer and non-oncology indications. This agreement was terminated in 2013.
Flt3 mutations are among the most common mutations in acute myeloid leukaemia due to internal tandem duplication of Flt3. The presence of this mutation is a marker of adverse outcome.
| Quizartinib is a small molecule with potential anticancer activity. Quizartinib is a selective inhibitor of class III receptor tyrosine kinases, including FMS-related tyrosine kinase 3 (FLT3/STK1), stem cell factor receptor (SCFR / KIT), colony-stimulating factor 1 receptor (CSF1R/FMS) and platelet-derived growth factor receptors (PDGFRs .) Able to inhibition of ligand-independent cell proliferation and apoptosis. Mutations in FLT3 are the most frequent genetic alterations in acute myeloid leukemia (AML) and occur in approximately 30% of cases of AML. | |
| Quizartinib представляет собой малую молекулу с потенциальной противораковой активностью. Quizartinib является селективным ингибитором класса III рецепторов тирозин киназ, в том числе FMS-связанных тирозинкиназы 3 (FLT3/STK1), фактор стволовых клеток рецепторов (SCFR / KIT), колониестимулирующий фактор 1 рецепторов (CSF1R/FMS) и тромбоцитарный рецепторов фактора роста (PDGFRs). Способен к торможению лиганд-независимой клеточной пролиферации и апоптоза. Мутации в FLT3 являются наиболее частыми генетическими изменениями в остром миелобластном лейкозе (ОМЛ) и встречаются примерно в 30% случаев ОМЛ. |
Mechanism
Specifically, Quizartinib selectively inhibits class III receptor tyrosine kinases, including FMS-related tyrosine kinase 3 (FLT3/STK1), colony-stimulating factor 1 receptor (CSF1R/FMS), stem cell factor receptor (SCFR/KIT), and platelet derived growth factor receptors (PDGFRs).
Mutations cause constitutive action of Flt3 leading to resulting in inhibition of ligand-independent leukemic cell proliferation and apoptosis.
Clinical trials
It had good results in a phase II clinical trial for refractory AML – particularly in patients who went on to have a stem cell transplant.[2]
………………………..
WO 2007109120 COMPD B1
EXAMPLE 3: PREPARATION OF N-(5-TERT-BUTYL-ISOXAZOL-3-YL)-N’-{4-[7-(2- MORPHOLIN-4-YL-ETHOXY)IMIDAZO[2,1 -B3[1 ,3]BENZOTHIAZOL-2-YL]PHENYL}UREA [Compound B1]
[00426] A. The intermediate 2-amino-1,3-benzothiazol-6-ol was prepared according to a slightly modified literature procedure by Lau and Gompf. J. Org. Chem. 1970, 35, 4103-4108. To a stirred solution of thiourea (7.6 g, 0.10 mol) in a mixture of 200 ml_ ethanol and 9 ml_ concentrated hydrochloric acid was added a solution of 1 ,4-benzoquinone (21.6 g, 0.20 mol) in 400 mL of hot ethanol. The reaction was stirred for 24 hours at room temperature and then concentrated to dryness. The residue was triturated with hot acetonitrile and the resulting solid was filtered and dried.
[00427] The free base was obtained by dissolving the hydrochloride salt in water, neutralizing with sodium acetate, and collecting the solid by filtration. The product (2-amino-1 ,3-benzothiazol-6-ol) was obtained as a dark solid that was pure by LCMS (M+H = 167) and NMR. Yield: 13.0 g (78 %). NMR (DMSOd6) £7.6 (m, 2H ), 6.6 (d, 1H).
[00428] B. To prepare the intermediate 2-(4-nitrophenyl)imidazo[2,1- b][1 ,3]benzothiazoI-7-ol, 2-amino-1 ,3-benzothiazol-6-ol, (20.0 g, 0.12 mol) and 2-bromo-4′-nitroacetophenone (29.3 g, 0.12 mol) were dissolved in 600 mL ethanol and heated to reflux overnight. The solution was then cooled to 00C in an ice-water bath and the product was collected by vacuum filtration. After drying under vacuum with P2O5 , the intermediate (2-(4- nitrophenyl)imidazo[2,1-_D][1,3]benzothiazol-7-ol) was isolated as a yellow solid. Yield: 17.0 g (46 %) NMR (DMSO-CT6) δ 10 (s, 1 H), 8.9 (s, 1H), 8.3 (d, 2H), 8.1 (d, 2H), 7.8 (d, 1 H), 7.4 (s, 1 H), 6.9 (d, 1 H). [00429] C. To make the 7-(2-morpholin-4-yl-ethoxy)-2-(4-nttro- phenyl)imidazo[2,1-£>][1 ,3]benzothiazo!e intermediate: 2-(4- nitrophenyl)imidazo[2,1-jb][1 ,3]benzothiazol-7-ol, (3.00 g, 9.6 mmol) was suspended in 100 mL dry DMF. To this mixture was added potassium carbonate (4.15 g, 30 mmol, 3 eq), chloroethyl morpholine hydrochloride (4.65 g, 25 mmol, 2.5 eq) and optionally tetrabutyl ammonium iodide (7.39 g, 2 mmol). The suspension was then heated to 900C for 5 hours or until complete by LCMS. The mixture was cooled to room temperature, poured into 800 mL water, and allowed to stand for 1 hour. The resulting precipitate was collected by vacuum filtration and dried under vacuum. The intermediate, (7-(2- morpholin-4-yl-ethoxy)-2-(4-nitro-phenyl)imidazo[2,1-jb][1 ,3]benzothiazole) was carried on without further purification. Yield: 3.87 g (95 %) NMR (DMSO-Cf6) δ 8.97 (s, 1 H), 8.30 (d, 2H), 8.0 (d, 2H), 7.9 (d, 1 H), 7.7 (s, 1 H), 7.2 (d, 1 H), 4.1 (t, 2H), 5.6 (m, 4H), 2.7 (t, 2H).
[00430] D. To make the intermediate 7-(2-morpholin-4-yl-ethoxy)-2-(4- amino-phenyl)!midazo[2, 1 -b][1 ,3]benzothiazole: To a suspension of 7-(2- morpholin-4-yl-ethoxy)-2-(4-nitro-phenyl)imidazo[2,1-ib][1 ,3]benzothiazole (3.87g, 9.1 mmol) in 100 ml_ isopropyl alcohol/water (3:1 ) was added ammonium chloride (2.00 g, 36.4 mmol) and iron powder (5.04 g, 90.1 mmol). The suspension was heated to reflux overnight with vigorous stirring, completion of the reaction was confirmed by LCMS. The mixture was filtered through Celite, and the filtercake was washed with hot isopropyl alcohol (150 ml_). The filtrate was concentrated to approximately 1/3 of the original , volume, poured into saturated sodium bicarbonate, and extracted 3 times with dichloromethane. The combined organic phases were dried over MgSO4 and concentrated to give the product as an orange solid containing a small amount (4-6 %) of starting material. (Yield: 2.75 g 54 %). 80% ethanol/water may be used in the place of isopropyl alcohol /water — in which case the reaction is virtually complete after 3.5 hours and oniy traces of starting material are observed in the product obtained. NMR (DMSO-d6) δ 8.4 (s, 1 H), 7.8 (d, 1 H), 7.65 (d, 1 H), 7.5 (d, 2H), 7.1 (d, 1 H), 6.6 (d, 2H), 4.1 (t, 2H)1.3.6 (m, 4H), 2.7 (t, 2H).
[00431] E. A suspension of 7-(2-morpholin-4-yl-ethoxy)-2-(4-amino- phenyl)imidazo[2,1-b][1 ,3]benzothiazole (4.06 g, 10.3 mmol) and 5-tert- butylisoxazole-3-isocyanate (1.994 g, 12 mmol) in toluene was heated at 120 0C overnight. The reaction was quenched by pouring into a mixture of methylene chloride and water containing a little methanol and neutralized with saturated aqueous NaHCO3 solution. The aqueous phase was extracted twice with methylene chloride, the combined organic extracts were dried over MgSO4 and filtered. The filtrate was concentrated to about 20 ml volume and ethyl ether was added resulting in the formation of a solid. The precipitate was collected by filtration, washed with ethyl ether, and dried under vacuum to give the free base. Yield: 2.342 g (41 %) NMR (DMSO-Cf6) £9.6 (br, 1H), 8.9 (br, 1H), 8.61 (s, 1H), 7.86 (d, 1 H), 7.76 (d, 2H), 7.69 (d, 1 H), 7.51 (d, 2H), 7.18 (dd, 1H), 6.52 (s, 1H), 4.16 (t, 2H), 3.59 (t, 4H), 3.36 (overlapping, 4H), 2.72 (t, 2H), 1.30 (s, 9H). NMR (CDCI3) £9.3 (br, 1H), 7.84 (m, 4H), 7.59 (d, 2H), 7.49 (d, 1 H), 7.22 (d, 1 H), 7.03 (dd, 1 H)1 5.88 (s, 1 H), 4.16 (t, 2H), 3.76 (t, 4H), 2.84 (t, 2H), 2.61 (t, 4H), 1.37 (s, 9H).
[00432] F. For the preparation of the hydrochloride salt, N-(5-tert-butyl- isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2, 1 – b][1 ,3]benzothiazol-2-yl]phenyI}urea hydrochloride, the free base was dissolved in a mixture of 20 ml methylene chloride and 1 ml methanol. A solution of 1.0 M HCI in ethyl ether (1.1 eq.) was added dropwise, followed by addition of ethyl ether. The precipitate was collected by filtration or centrifugation and washed with ethyl ether to give the hydrochloride salt. Yield: 2.44 g (98 %) NMR (DMSO-d6) £11-0 (br, 1 H), 9.68 (s, 1H), 9.26 (s, 1H), 8.66 (s, 1 H), 7.93 (d, 1H), 7.78 (m, 3H), 7.53 (d, 2H), 7.26 (dd, 1H), 6.53 (S, 1 H), 4.50 (t, 2H), 3.97 (m, 2H), 3.81 (t, 2H), 3.6 (overlapping, 4H)13.23 (m, 2H)1 1.30 (s, 9H).
[00433] G. Alternatively, Compound B1 may be made by taking the intermediate from Example 4B and reacting it with chloroethyl morpholine hydrochloride under conditions described in Step C. [00434] H . Λ/-(5-tert-butyl-isoxazol-3-yl)-Λ/’-{4-[5-(2-morpholin-4-yl- ethoxy)imidazo[2,1-6][1 ,3]benzothiazol-2-yl]phenyl}urea hydrochloride, a compound having the general formula (I) where R1 is substituted on the 5 position of the tricyclic ring, was prepared in the manner described in Steps A- F but using the cyciization product 2-amino-benzothiazol-4-ol with 2-bromo-4′- nitroacetophenone in Step A. 1H NMR (DMSO-d6) δ 11.6 (br, 1 H)1 9.78 (br, 1H), 9.56 (br, 1 H), 8.64 (s, 1H)1 7.94 (d, 2H), 7.70 (s, 1H)1 7.56 (d, 2H), 7.45 (t, 1 H), 7.33 (d, 1H), 6.54 (s, 1 H), 4.79 (t, 2H), 3.87 (m, 6H), 3.60 (m, 2H), 3.34 (m, 2H)1 1.30 (s, 9H); LC-MS: ESI 561 (M+H)+. [Compound B11] [00435] I. N-(5-tert-butyl-isoxazol-3-yl)-N’-{4-[6-(2-morpholin-4-yl- ethoxy)imidazo[2,1-b][1 ,3]benzothiazol-2-yl]phenyl}urea hydrochloride [Compound B12] was also prepared by first preparing the benzothiazole starting material, 5 methoxy-benzothiazol-2yl~amine: [00436] To prepare the 5-methoxy-benzothiazol-2-ylamine starting material: To a suspension of (3-methoxy-phenyl)-thiourea (1.822g, 10 mmol) in CH2CI2 (20 ml_) at 0 0C was added dropwise a solution of bromine (1.76 g, 11 mmol) in 10 ml of trichloromethane over a period of thirty minutes. The reaction was stirred for 3 hours at room temperature then heated to 3 hours to reflux for one hour. The precipitate was filtered and washed with dichloromethane. The solid was suspended in saturated NaHCOsand extracted with CH2CI2. The extract was dried over MgSO4 and concentrated to give a white solid (1.716 g, 95%).
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WO 2011056939
N-(5-ieri-butyl- isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l-&][l,3]benzothiazol-2- yl]phenyl}urea (I), or a pharmaceutically acceptable salt, solvate, hydrate, or polymorph thereof. N-(5-ieri-Butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- / ][!, 3]benzo
N- (5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- &][l,3]benzo-thiazol-2-yl]phenyl}urea (I), or a pharmaceutically acceptable salt, solvate, hydrate, or polymorph thereof, comprising any one, two, three, four, five, six, seven of the steps of:
(A) converting 2-amino-6-alkoxybenzothiazole (II), wherein R1 is a suitable phenolic hydroxyl protecting ;
(II) (III)
(B) reacting 2-amino-6-hydroxybenzothiazole (III) with compound (IV), wherein X is a leaving group, to yield 2-(4-nitrophenyl)imidazo[2,l-b]benzothiazol-7-ol (V);
(C) reacting 2-(4-nitrophenyl)imidazo[2,l-b]benzothiazol-7-ol (V) with compound (VI), wherein X2 is a leaving group, to yield 7-(2-morpholin-4-yl-ethoxy)-2-(4- nitrophenyl)imidazo[ -b]benzothiazole (VII);
(D) reducing 7-(2-morpholin-4-yl-ethoxy)-2-(4-nitrophenyl)imidazo[2, 1- bjbenzothiazole (VII) to yield 7-(2-morpholin-4-yl-ethoxy)-2-(4- am
(E) converting 3-amino-5-£er£-butyl isoxazole (IX) to a 5-?er?-butylisoxazol-3- ylcarbamate derivative (X), wherein R2 is optionally substituted aryl, heteroaryl, alkyl, or cycloalkyl;
(IX) (X)
(F) reacting 7-(2-morpholin-4-yl-ethoxy)-2-(4-aminophenyl)imidazo[2,l- bjbenzothiazole (VIII) with a 5-£er£-butylisoxazol-3-ylcarbamate derivative (X) to yield N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- &][l,3]benzo-
(G) converting N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl- ethoxy)imidazo[2,l-&][l,3]benzo-thiazol-2-yl]phenyl}urea to an acid addition salt of N- (5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- b] [ 1 ,3]benzo-thiazol-2-yl]phenyl } urea.
[00128] In certain embodiments, provided herein are processes for the preparation of N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- &][l,3]benzo-thiazol-2-yl]phenyl}urea, or a pharmaceutically acceptable salt, solvate, hydrate, or polymorph thereof, as depicted in Scheme 1, wherein R1, R2, X1, and X2 are defined herein elsewhere. In specific embodiments, provided herein are processes for the preparation of N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl- ethoxy)imidazo[2,l-&][l,3]benzo-thiazol-2-yl]phenyl}urea (I), or a pharmaceutically acceptable salt, solvate, hydrate, or polymorph thereof, comprising any one, two, three, four, five, six, seven, of the Steps A, B, C, D, E, F, and G, as depicted in Scheme 1.
Scheme 1 :
A. Preparation of 2-amino-6-hydroxybenzothiazole
1. Example A-l[00252] To a 1-L 3-necked round bottom flask fitted with a condenser, heating mantle, and mechanical stirrer was charged aqueous hydrobromic acid (48%, 632 mL, 5.6 mol, 10 equiv). 2-Amino-6-methoxybenzothiazole (100 g, 0.55 mol, 1 equiv) was added to the above flask over 15 minutes. The reaction temperature was raised slowly to reflux (105-110 °C). A clear dark brown colored solution was observed at about 80 °C. The reflux was continued at 105-110 °C for about 4 hr. The progress of the reaction was monitored by HPLC. When 2-amino-6-methoxybenzothiazole was less than 2%, the reaction was substantially complete.
[00253] The reaction mass was cooled to 0-5 °C and at this point precipitation of a solid was observed. The mixture was maintained at 0-5 °C for 0.5 hr and filtered, and the cake was pressed to remove HBr. The wet cake was transferred to a 2-L round bottom flask fitted with a mechanical stirrer. Saturated aqueous sodium bicarbonate solution (-1500 mL) was added slowly at ambient temperature, whereupon considerable frothing was observed. The pH of the solution was found to be about 6.5 to 7. The mixture was stirred for 0.5 hr at ambient temperature and filtered. The filter cake was washed with water (500 mL), dried on the filter and then under vacuum at 30-35 °C for 10-12 hr to give the product 2-amino-6-hydroxybenzothiazole (82 g, 89% yield, HPLC purity = 99%). JH NMR (DMSO-if6, 500 MHz): δ 7.12 (d, 1H), 7.06 (S, 2H, NH2), 7.01 (d, 1H), 6.64 (dd, 1H); MS (m/z) = 167.1 [M+ + 1].
[00254] Table: Summary of Plant Batches
[00255] HPLC chromatographic conditions were as follows: The column used was XTerra RP8, 250 X 4.6 mm, 5μ or equivalent. Mobile Phase A was buffer, prepared by mixing 3.48 g of dipotassium hydrogen phosphate in 1.0 L of water, and adjusting the pH to 9.0 with phosphoric acid. Mobile Phase B was methanol. The flow rate was 1.0 mL/minute. Detection was set at UV 270 nm. The injection volume was 20 μΐ^, and the sample was diluted with a diluent (Mobile Phase A : Mobile Phase B = 70:30). Test solution was prepared by weighing accurately about 25 mg of sample and transferring it into a 100 mL volumetric flask, dissolving with 20-30 mL of diluent, making up the volume to the mark with diluent, and mixing. The HPLC was performed by separately injecting equal volumes of blank and test solution, and recording the chromatogram for all injections. The purity was calculated by area normalization method.
[00256] Table: HPLC Method
2. Example A-2
[00257] 2-Amino-6-methoxybenzothiazole was reacted with hot aqueous HBr at a temperature of >70 °C for about 3 hours and then the clear solution was cooled to ambient temperature overnight. The precipitated solids were collected, dissolved in hot water and the pH was adjusted to between 4.5-5.5. The resultant solids were collected, dried and re-crystallized from isopropanol. Second crop material was collected. The solids were vacuum dried to give 2-amino-6-hydroxybenzothiazole.
[00258] The reaction progress was monitored by thin layer chromatography (TLC). The product was isolated as a white solid, with 99.4% purity (HPLC area %). JH NMR (300 MHz, DMSO-if6) was collected, which conformed to structure.
3. Example A-3
[00259] A 22-L 3-neck round bottom flask was equipped with a mechanical agitator, thermocouple probe, a reflux condenser, and a heating mantle. The flask was charged with hydrobromic acid (14 L, 123.16 mol, 13.10 equiv). Heating was initiated and 2- amino-6-methoxybenzothiazole was added (1.7 kg, 9.4 mol, 1.00 equiv) over 10 minutes with stirring. The heating of the reaction mixture was continued to reflux, and maintained (>107 °C) for approximately 5 hours. The reaction mixture turned into a clear solution between 75 °C and 85 °C. The reaction progress was monitored by TLC until no starting material was observed (A -0.5 mL reaction mixture aliquot was diluted with -0.5 mL water as a clear solution, neutralized with sodium acetate to pH -5 and extracted with 1 mL dichloromethane. The organic layer was spotted: 5%
methanol/dichloromethane; Rf (product) = 0.35; Rf (starting material) = 0.40).
[00260] The reaction mixture was cooled to – 20 °C (overnight). White solids precipitated. The solids were filtered on a polypropylene filter and pressed to remove as much hydrobromic acid from the solids as possible to facilitate the subsequent pH adjustment step. The slightly wet crude product was dissolved in hot (50 °C) water (5 L). The clear solution was filtered to remove any insoluble material present, and the solids were washed with 50 °C water. The filtrate was cooled to 10 °C. The cooled filtrate was neutralized with sodium acetate (- 1.0 kg) to pH 4.5 to 5.5 with vigorous stirring. A thick white solid precipitated. The solids were collected by filtration, and washed with cool (-10 °C) water (2 x 1.0 L) and air dried.
[00261] The wet crude product was slurried in hot (50 °C) isopropanol (3 L) briefly and allowed to stand in a cool room (-5 °C) overnight. The solids were collected by filtration and washed with methyl ferf-butylether (2 x 500 mL). The solids were dried in a vacuum oven overnight (<30 mm Hg) at 30 °C (first crop). Yield: 1068 g (68%), white solid. HPLC: 99.4% (area). JH NMR (300 MHz, DMSO- ) conformed to structure.
[00262] The organic filtrate was collected in a total volume of 1.0 L, cooled to 10 °C. The off-white solids were precipitated and collected by filtration. The solids were dried in a vacuum oven overnight (<30 mm Hg) at 30 °C (second crop). Yield: 497 g (32%), off-white solid. HPLC: 99.8% (area).
[00263] The overall yield combining the first crop and the second crop was 1565 g, (99%).
B. Preparation of 2-(4-nitrophenyl)imidazo[2,l-b]benzothiazol-7-ol
1. Example B-l[00264] A 3-L 3-neck round bottom flask fitted with a condenser, a heating mantle, and a mechanical stirrer was charged with H-butanol (1.5 L), followed by 2-amino-6- hydroxybenzothiazole (75 g, 0.45 mol, 1.0 equiv), 2-bromo-4′-nitroacetophenone (121 g, 0.50 mol, 1.1 equiv), and sodium bicarbonate (41.6 g, 0.50 mol, 1.0 equiv). The reaction temperature was gradually raised to reflux and maintained at reflux (110-115 °C) for 2-3 hr. During the temperature increase, the reaction mass turned into a clear solution and then immediately changed into an orange colored suspension at 65-75 °C. The progress of the reaction was monitored by HPLC analysis every 1 hr (reaction mass sample was submitted to QC). When the level of 2-amino-6-hydroxybenzothiazole was less than 2%, the reaction was substantially complete.
[00265] The reaction mass was slowly cooled to 50-60 °C and then further cooled to 0-5 °C and stirred for 15 min. The precipitated solids were collected by filtration, and dried on the filter. The wet cake was transferred in to a 1-L round bottom flask, and water (600 mL) was added. The suspension was stirred for 0.5 hr and filtered, and the solid was dried on the filter. The wet cake was again taken in to a 1-L round bottom flask and stirred with acetone (200 mL). The slurry was filtered and washed with acetone (2 X 100 mL), and the solid was dried on the filter, unloaded and further dried in a vacuum oven at ambient temperature to give the product 2-(4-nitrophenyl)imidazo[2,l- b]benzothiazol-7-ol (V) (120 g, 85.7% yield, HPLC purity = 98.7%). JH NMR (DMSO- d6, 500 MHz): δ 9.96 (s, 1H, OH), 8.93 (s, 1H), 8.27 (d, 2H), 8.06 (d, 2H), 7.78 (d, 1H), 7.38 (d, 1H), 6.97 (dd, 1H); MS (m/z) = 312 [M+ + 1].
[00266] Table: Summary of Plant Batches
* Input of 2-amino-6-hydroxybenzothiazole (III)
[00267] HPLC chromatographic conditions were as follows: The column used was XTerra RP8, 250 X 4.6 mm, 5μ or equivalent. Mobile Phase A was buffer, prepared by mixing 3.48 g of dipotassium hydrogen phosphate in 1.0 L of water, and adjusting the H to 9.0 with phosphoric acid. Mobile Phase B was methanol. The flow rate was 1.0 mL/minute. Detection was set at UV 235 nm. The injection volume was 10 μΐ^. The blank was prepared by transferring 200 μΐ. of DMSO and 200 μΐ. of 2M NaOH into a 10 mL volumetric flask, making up the volume to the mark with methanol, and mixing. The test solution was prepared by weighing accurately about 10 mg of sample and transferring it into a 50 mL volumetric flask, dissolving with 1 mL of DMSO and 1 mL of 2M NaOH, sonicating to dissolve, making up the volume to the mark with methanol, and mixing. The HPLC was performed by separately injecting equal volumes of blank and test solution, and recording the chromatogram for all injections. The purity was calculated by area normalization method.
[00268] Table: HPLC Method
2. Example B-2
[00269] A 50-L 3-neck round bottom flask was equipped with a mechanical agitator, a thermocouple probe, a reflux condenser, and a heating mantle. The flask was charged with 2-amino-6-hydroxybenzothiazole (1068 g, 6.43 mol, 1.0 equiv) and ethanol (200 proof, 32.0 L), and the suspension was stirred for 10 minutes. 2-Bromo-4- nitroacetophenone (1667 g, 6.49 mol, 1.01 equiv) was added in one portion. The reaction mixture was heated to reflux (78 °C). The reflux was maintained for approximately 25 hours, resulting in a yellow suspension. The reaction progress was monitored by TLC (20% methanol/ethyl acetate; Rf (product) = 0.85; Rf (starting material) = 0.30). TLC indicated -50% 2-amino-6-hydroxybenzothiazole after 24 hours of reflux. Tetrabutylammonium iodide (10 g) was added and reflux was maintained for an additional 12 hours. TLC indicated -50% starting material still present. Coupling was seen to occur at both the thiazole and the amine.
[00270] The reaction mixture was cooled to 0-5 °C. The solids were collected by filtration, and the yellow solid was washed with ethanol (200 proof, 2 X 1.0 L) and diethyl ether (2 X 1.5 L). The solids were dried in a vacuum oven (<10 mm Hg) at 40 °C. Yield: 930 g (46%), yellow solid. HPLC: 99.5% (area). JH NMR (300 MHz, DMSO-i¾) conformed to structure.
3. Example B-3
[00271] The reaction of Step B was carried out on multiple runs, varying solvents, temperature, and base. The results were summarized in the table below. The product (V) was isolated as yellow or green solids, with 1H NMR consistent with the structure and a purity of greater than about 98% by HPLC analysis.
[00272] Table: Reaction Condition Screening
TBAI = Tetrabutylammonium Iodide
C. Preparation of 7-(2-morpholin-4-yl-ethoxy)-2-(4- nitrophenyl)imidazo[2, 1 -bjbenzothiazole
1. Example C-l
[00273] To a 2000-L glass-lined (GL) reactor was charged DMF (298 kg), and the agitator was started. Under a nitrogen blanket, the reactor was charged with 2-(4- nitrophenyl)imidazo[2,l-&]benzothiazol-7-ol (36.8 kg, 118.2 mol, 1.0 equiv), 4-(2- chloroethyl)morpholine hydrochloride (57.2-66.0 kg, 307.3-354.6 mol, 2.6-3.0 equiv), tetrabutylammonium iodide (8.7 kg, 23.6 mol, 0.2 equiv) and potassium carbonate (49.0 kg, 354.6 mol, 3.0 equiv). The resulting yellow suspension was heated and stirred at 90 + 5 °C for at least 15 minutes, then the temperature was increased to 110 + 5 °C. The mixture was stirred for at least 1 hour and then sampled. The reaction was deemed complete if 2-(4-nitrophenyl) imidazo[2,l-&]benzothiazol-7-ol was <1% by HPLC. If the reaction was not complete, the heating was continued and the reaction sampled. If the reaction was incomplete after 6 hours, additional 4-(2-chloroethyl)morpholine hydrochloride may be charged. In general, additional charges of 4-(2- chloroethyl)morpholine hydrochloride had not been necessary at the given scale.
[00274] The reactor was cooled to 20 + 5 °C and charged with water (247 kg) and acetone (492 kg). The mixture was agitated at 20 + 5 °C for at least 1 hour. The product (VII) was isolated by filtration or centrifuge, and washed with water and acetone, and then dried in a vacuum oven at 45 °C to constant weight to give a yellow solid (46.2 kg, 92% yield, HPLC purity = 97.4% by area). JH NMR (300 MHz, DMSO- ) conformed to structure.
2. Example C-2
[00275] 2-(4-Nitrophenyl)imidazo[2, l-b]benzothiazol-7-ol, 4-(2-chloroethyl)- morpholine hydrochloride, potassium carbonate, and tetrabutylammonium iodide were added to N,N-dimethylformamide forming a yellow suspension that was heated at a temperature of >50 °C for over 3 hours. The reaction was cooled and the solids were collected, slurried into water, filtered, slurried into acetone, filtered and washed with acetone to give yellow solids that were dried under vacuum to give 7-(2-morpholin-4-yl- ethoxy)-2-(4-nitrophenyl)imidazo[2,l-b]benzothiazole.
[00276] The reaction progress was monitored by thin layer chromatography (TLC). The product was isolated as a yellow solid, with 99% purity (HPLC area %), and a water content of 0.20%. Infrared (IR) spectrum was collected, which conformed to structure.
3. Example C-3
[00277] A 50-L 3-neck round bottom flask was equipped with a mechanical agitator, a thermocouple probe, a drying tube, a reflux condenser, and a heating mantle. The flask was charged with 2-(4-nitrophenyl)imidazo [2,l-&]benzothiazol-7-ol (1.770 kg, 5.69 mol, 1.0 equiv), N,N-dimethylformamide (18.0 L), 4-(2-chloroethyl)morpholine hydrochloride (2.751 kg, 14.78 mol, 2.6 equiv), potassium carbonate (2.360 kg, 17.10 mol, 3.0 equiv), and tetrabutylammonium iodide (0.130 kg, 0.36 mol, 0.06 equiv) with stirring. The resulting yellow suspension was heated to about 90 °C to 95 °C, maintaining the temperature for approximately 5 hours. The reaction was monitored by TLC until no starting material was observed (20% methanol / ethyl acetate; Rf (product) = 0.15; Rf (starting material) = 0.85).
[00278] The reaction mixture was cooled to -10 °C, and the yellow solids were collected by filtration on a polypropylene filter pad. The solids were slurried in water (2 X 5 L) and filtered. The crude wet product was slurried in acetone (5 L), filtered, and the solids were rinsed with acetone (2 X 1.5 L). The solids were dried in a vacuum oven (<10 mm Hg) at 45 °C. Yield: 2.300 kg (95%), yellow solid. TLC: R/ = 0.15 (20% methanol / EtOAc). HPLC: 95.7% (area). JH NMR (300 MHz, DMSO-i¾) conformed to the structure.
[00279] Table: Yields from multiple batch runs
4. Example C-4
[00280] To a reactor were added 2-(4-nitrophenyl)imidazo [2,l-&]benzothiazol-7-ol (1.0 kg), 4-(2-chloroethyl)morpholine hydrochloride (1.6 kg), tetrabutylammonium iodide (0.24 kg), and potassium carbonate (1.3 kg, anhydrous, extra fine, hydroscopic). N,N-Dimethylformamide (DMF) (8.6 L) was added into the reactor. The DMF used had water content of no more than 0.05% w/w. The mixture was stirred for between 15 and 30 minutes to render a homogeneous suspension, which was heated to between 85 °C and 95 °C and stirred at between 85 °C and 95 °C for 15 to 30 minutes. The mixture was then heated to between 105 °C and 120 °C and stirred at between 105 °C and 120 °C (e.g. , 115 °C) until the reaction was complete (as determined by HPLC to monitor the consumption of starting material). In some embodiments, if necessary (e.g. , if after 6 hours the reaction was not complete as indicated by HPLC analysis), additional 4-(2- chloroethyl)morpholine hydrochloride (0.03 kg) may be added and the reaction mixture stirred at between 105 °C and 120 °C (e.g. , 115 °C) until reaction completion.
[00281] The reaction mixture was cooled to between 20 °C and 30 °C (e.g. , over a period of 3 hours). To another reactor was added deionized water (7.6 L) and acetone (15 L). The mixture of water and acetone was then added into the reaction mixture while maintaining the temperature at between 20 °C and 30 °C. The mixture was then stirred for 1 to 2 hours at a temperature of between 20 °C and 30 °C. The mixture was filtered, and the solid was washed with deionized water (e.g. , about 45x deionized water) until pH of washes was below 8. The solid was then washed with acetone (9.7 L). The solid was dried under vacuum at a temperature of less than 50 °C until the water content by Karl-Fischer was less than 0.30% w/w and TGA curve showed a mass loss of less than 0.30% w/w at about 229 °C (sampling approximately every 6 hours). The desired product was obtained in about 89% yield having about 99% purity by HPLC.
5. Example C-5
[00282] To a reactor were added 2-(4-nitrophenyl)imidazo [2, l-&]benzothiazol-7-ol (1.0 kg), 4-(2-chloroethyl)morpholine hydrochloride (1.6 kg), and potassium carbonate (1.3 kg, anhydrous, extra fine, hydroscopic). N,N-Dimethylformamide (DMF) (8.6 L) was added into the reactor. The DMF used had water content of no more than 0.05% w/w. The mixture was stirred for between 15 and 30 minutes to render a homogeneous suspension, which was heated to between 95 °C and 120 °C (e.g. , between 100 °C and 105 °C) and stirred at between 95 °C and 120 °C (e.g. , 105 °C) until the reaction was complete (as determined by HPLC to monitor the consumption of starting material). In some embodiments, if necessary (e.g. , if after 6 hours the reaction was not complete as indicated by HPLC analysis), additional 4-(2-chloroethyl)morpholine hydrochloride (0.03 kg) and potassium carbonate (0.024 kg) may be added and the reaction mixture stirred at between 100 °C and 120 °C (e.g. , 105 °C) until reaction completion.
[00283] The reaction mixture was cooled to between 60 °C and 70 °C over a period of at least 60 minutes. Industrial water (6 L) was added to the reactor. The reaction mixture was cooled to between 20 °C and 30 °C. Acetone (6 L) was added to the reactor. The mixture was stirred for 1 to 2 hours at a temperature of between 20 °C and 30 °C. The mixture was filtered, and the solid was washed with industrial water (e.g. , about 45 x industrial water) until pH of washes was below 8. The solid was then washed with acetone (9.7 L). The solid was dried under vacuum at a temperature of less than 50 °C, until the water content by Karl-Fischer was less than 0.30% w/w and TGA curve showed a mass loss of less than 0.30% w/w at about 229 °C (sampling approximately every 6 hours).
6. Example C-6
[00284] To a reactor is added 2-(4-nitrophenyl)imidazo [2, l-&]benzothiazol-7-ol (1.0 kg), 4-(2-chloroethyl)morpholine hydrochloride (1.6 kg), and potassium carbonate (1.3 kg, anhydrous, extra fine, hydroscopic). N,N-Dimethylformamide (DMF) (8.6 L) is added into the reactor. The DMF has a water content of no more than 0.05% w/w. The mixture is stirred for between 15 and 30 minutes to render a homogeneous suspension, which is heated to between 95 °C and 120 °C (e.g. , between 100 °C and 105 °C) and stirred at between 95 °C and 120 °C (e.g. , 105 °C) until the reaction is complete (as determined by HPLC to monitor the consumption of starting material). In some embodiments, if necessary (e.g. , if after 6 hours the reaction is not complete as indicated by HPLC analysis), additional 4-(2-chloroethyl)morpholine hydrochloride (0.03 kg) and potassium carbonate (0.024 kg) may be added and the reaction mixture stirred at between 100 °C and 120 °C (e.g. , 105 °C) until reaction completion.
[00285] The reaction mixture is cooled to between 20 °C and 30 °C (e.g. , over a period of 3 hours). To another reactor is added deionized water (7.6 L) and acetone (15 L). The mixture of water and acetone is then added into the reaction mixture while maintaining the temperature at between 20 °C and 30 °C. The mixture is then stirred for 1 to 2 hours at a temperature of between 20 °C and 30 °C. The mixture is filtered, and the solid is washed with deionized water (e.g. , about 45x deionized water) until pH of washes is below 8. The solid is then washed with acetone (9.7 L). The solid is dried under vacuum at a temperature of less than 50 °C until the water content by Karl-Fischer is less than 0.30% w/w and TGA curve shows a mass loss of less than 0.30% w/w at about 229 °C (sampling approximately every 6 hours). D. Preparation of 7-(2-morpholin-4-yl-ethoxy)-2-(4- aminophenyl)imidazo [2, 1 -bjbenzothiazole
[00286] To a 200-L high pressure (HP) reactor was charged a slurry of 7-(2- morpholin-4-yl-ethoxy)-2-(4-nitrophenyl)imidazo [2,l-&]benzothiazole (VII) (7.50 kg, 17.7 mol, 1.0 equiv) in methanol (30 kg). The container was rinsed with additional methanol (10 kg) and the rinse was charged to the reactor. The reactor was then charged with THF (67 kg) and methanol (19 kg). The contents were agitated and the reactor was flushed with nitrogen by alternating nitrogen and vacuum. Vacuum was applied to the reactor and Raney Ni catalyst (1.65 kg, 0.18 wt. equiv) was charged through a sample line. Water (1 kg) was charged through the sample line to rinse the line. The reactor was flushed with nitrogen by alternating nitrogen and vacuum. The reactor was then vented and heated to 50 °C. The reactor was closed and pressurized with hydrogen gas to 15 psi keeping the internal temperature below 55 °C. The reactor was vented and re- pressurized a total of 5 times, then pressurized to 150 psi with hydrogen gas. The contents were agitated at 50 °C for at least 4 hours. At this point a hydrogen uptake test was applied: The reactor was re-pressurized to 150 psi and checked after 1 hour. If a pressure drop of more than 5 psi was observed, the process was repeated. Once the pressure drop remained < 5 psi, the reactor was vented and sampled. The reaction was deemed complete when 7-(2-morpholin-4-yl-ethoxy)-2-(4-nitrophenyl)imidazo [2,1- 6]benzothiazole (VII) was < 0.5% by HPLC.
[00287] The reactor was flushed with nitrogen as shown above. The 200-L HP reactor was connected to the 2000-L GL reactor passing through a bag filter and polish filter. The bag filter and polish filter were heated with steam. The 200-L HP reactor was pressurized (3 psi nitrogen) and its contents were filtered into the 2000-L reactor. The filtrates were hot. The 200-L reactor was vented and charged with THF (67 kg) and methanol (59 kg), the reactor agitated, and filtered into the 2000-L GL reactor.
[00288] A total of 6 reductions (46.2 kg processed) were carried out and the combined batches were concentrated by vacuum distillation (without exceeding an internal temperature of 40 °C) to a volume of -180 L. The reactor was cooled to 20 °C and charged with heptane (250 kg) and again vacuum distilled to a volume of -180 L. The reactor was charged with heptane (314 kg) and agitated at 20 °C for at least 1 hour, and then the product was isolated by centrifugation or collection on a Nutsche filter, washing with heptanes (2-5 kg per portion for centrifugation, followed by a 10-20 kg heptanes rinse of the reactor; or 94 kg for Nutsche filtration, rinsing the reactor first). The cake was blown dry, transferred to a vacuum oven and dried to constant weight maintaining a temperature < 50 °C to give the desired product (VIII) (34.45 kg, 80% yield, HPLC purity = 97.9%).
2. Example D-2
[00289] 7-(2-Morpholin-4-yl-ethoxy)-2-(4-nitrophenyl)imidazo[2,l-b]benzothiazole was dissolved into methanol and THF and placed in a hydrogenator. Raney nickel was added and the vessel was pressurized with hydrogen and stirred for >24 hours. The reaction mixture was concentrated to a thick paste and diluted with methyl ferf-butyl ether. The resulting solids were filtered and washed with methyl ferf-butyl ether and dried under vacuum to give 7-(2-morpholin-4-yl-ethoxy)-2-(4-aminophenyl) imidazo [2, 1 -bjbenzothiazole.
[00290] The reaction progress was monitored by thin layer chromatography (TLC). The product was isolated as a yellow solid, with 99% purity (HPLC area %). IR was collected, which conformed to structure.
3. Example D-3
[00291] Into a 5-gallon autoclave, 7-(2-morpholin-4-yl-ethoxy)-2-(4-nitrophenyl) imidazo[2,l-&]benzothiazole (580 g, 1.37 mol, 1.0 equiv), THF (7.5 L), methanol (7.5 L, AR) and -55 g of decanted Raney nickel catalyst were added. The reaction vessel was purged with nitrogen (3 X 50 psi) and hydrogen (2 X 50 psi), with stirring briefly under pressure and then venting. The autoclave was pressurized with hydrogen (150 psi). The mixture was stirred and the hydrogen pressure was maintained at 150 psi for over 24 hours with repressurization as necessary. The reaction progress was monitored by TLC (10% methanol / chloroform with 1 drop ammonium hydroxide; Rf (product) 0.20; Rf (SM) 0.80). The reaction was substantially complete when the TLC indicated no starting material present, typically after 24 hours of stirring at 150 psi. The hydrogenation was continued at 150 psi for a minimum of 4 hours or until completion if starting material was still present after the initial 4 hours.
[00292] The reaction mixture was filtered through a Buchner funnel equipped with a glass fiber filter topped with a paper filter. Unreacted starting material was removed together with the catalyst. The filtrate was concentrated to a total volume of 0.5 L, and the residue was triturated with methyl ferf-butyl ether (0.5 L). The resultant solids were collected by filtration, and washed with methyl ferf-butyl ether (0.3 L) (first crop).
[00293] The filtrate was concentrated to dryness and the residue was diluted with methyl ferf-butyl ether (2 L). The resultant solids were collected by filtration, washing with methyl ferf-butyl ether (0.5 L) (second crop).
[00294] The solids were dried in a vacuum oven (<10 mm Hg) at 25 °C. Yield: 510 g (95%), beige solid. TLC: R/ 0.2 (10% methanol / chloroform with one drop of ammonium hydroxide). HPLC: 99.0% (area). JH NMR (300 MHz, DMSO-i¾) conformed to the structure.
[00295] Table: Yields from multiple batch runs
4. Example D-4
[00296] The reaction of Step D was carried out in multiple runs under various conditions, such as, e.g. , varying catalyst loading, concentration of reactant, reaction temperature, and/or workup procedures. The results are summarized in the table below.
Description Run # l Run # 2 Run # 3 Run # 4 Run # 5Rxn Temp (°C) RT RT RT RT RT
Rxn Time (Hr) 24 hr 24 hr 24 hr 24 hr 24 hr
Filtered the Filtered the solution
Filtered the Filtered the Filtered the
solution through through celite. The solution through solution through solution through
celite, washed celite filter cake celite, celite, celite,
with THF, refluxed in THF concentrated, concentrated, concentrated,
concentrated, washed with hot solvent exchanged solvent exchanged solvent exchanged
Work Up solvent exchanged THF, concentrated, with heptane, with heptane, with heptane,
with heptane, solvent exchanged stirred the solids stirred the solids stirred the solids
stirred the solids with heptane, stirred and filtered and filtered and filtered
and filtered the solids and washed with washed with washed with
washed with filtered washed with heptane heptane heptane
heptane heptane
Produce (VIII) 1.9 g 3.88 g 1.11 g 2.6 g 4.4 g
Yield 88% 83.4% 56 94.6%
HPLC purity 95.6% 77.5% 91% 93.8%
5. Example D-5
[00297] To a pressure reactor under nitrogen atmosphere was added a slurry of Raney Nickel in water (0.22 kg) (e.g. about 0.14 kg dry catalyst in water) and the charging line was rinsed with deionized water (0.13 L). To another reactor (Reactor B) were added methanol (5.05 L) and 7-(2-morpholin-4-yl-ethoxy)-2-(4-nitrophenyl)imidazo [2, 1- &]benzothiazole (1.0 kg), and the mixture was stirred for between 15 and 30 minutes to render a homogenous suspension. The suspension was transferred to the pressure reactor. Reactor B was washed with methanol (4.88 L) and the wash was transferred to the pressure reactor. Tetrahydrofuran (10.1 L) was added to the pressure reactor.
Hydrogen was charged to the pressure reactor to a pressure of between 2.0 bar and 3.0 bar. The reactor was heated to a temperature of between 45 °C and 55 °C. Hydrogen was then charged to the pressure reactor to a pressure of between 6.0 bar and 7.0 bar. The mixture was stirred at a temperature of between 45 °C and 55 °C (e.g. , 50 °C), while maintaining the hydrogen pressure between 6.0 bar and 7.0 bar until reaction completion (as determined by HPLC to monitor the consumption of starting material).
[00298] The mixture was filtered while maintaining the temperature at between 35 °C and 50 °C. The pressure reactor and the filter were washed with a mixture of THF (10.1 L) and methanol (9.93 L) preheated to a temperature of between 45 °C and 55 °C (e.g. , 50 °C). The combined filtrate was concentrated to a volume of between 2.4 L and 2.8 L under vacuum at a temperature of no more than 40 °C, and a precipitate was formed. Methanol (7.5 L) was added. The slurry was cooled to a temperature of between 5 °C and -5 °C (e.g. , over 3 hours) and stirred for at least 1 hour (e.g. , for 3 hours) while maintaining the temperature at between 5 °C and -5 °C. The suspension was filtered. The solid was washed with methanol (2 X 1.2 L). The solid was then dried under vacuum at a temperature of less than 50 °C until the methanol and THF contents were each less than 3000 ppm as analyzed by GC (e.g. , less than 1500 ppm). The desired product was obtained in about 88.5% yield having about 99% purity by HPLC.
E. Preparation of phenyl 5-£er£-butylisoxazol-3-ylcarbamate
[00299] The jacket temperature of a 200-L glass-lined (GL) reactor was set to 20 °C. To the reactor was charged 5-ieri-butylisoxazole-3-amine (15.0 kg, 107.0 mol, 1.0 equiv), then K2C03 (19.5 kg, 141.2 mol, 1.3 equiv) and anhydrous THF (62 kg).
Agitation was started and then phenyl chloroformate (17.6 kg, 112.4 mol, 1.05 equiv) was charged. The charging line was rinsed with additional anhydrous THF (5 kg). The reaction was agitated at 20 + 5 °C for at least 3 hours then sampled. The reaction was deemed complete if 5-£er£-butylisoxazole-3-amine was < 2% by HPLC. If the reaction was not complete after 6 hours, additional K2CO3 and phenyl chloroformate may be added to drive the reaction to completion.
[00300] Once complete, the reaction was filtered (Nutsche). The filter was rinsed with THF (80 kg). The filtrate was vacuum distilled without exceeding an internal temperature of 40 °C until -50 L remained. Water (188 kg) and ethanol (45 L) were charged, and the mixture was agitated for at least 3 hours with a jacket temperature of 20 °C. The resulting solid was isolated by centrifugation or collection on a Nutsche filter, rinsed with water (2-5 kg for each centrifugation portion; 30 kg for Nutsche filtration) and blow-dried. The solid was then dried to constant weight in a vacuum oven (45 °C) to give the desired product (19.4 kg, 92% yield, HPLC purity = 97.4%). On an 800 g scale, 1559 g of the desired product (98% yield) was obtained with a 99.9% HPLC purity. JH NMR (DMSO-i¾) δ 11.17 (s, 1H); 7.4 (m, 2H); 7.2 (m, 3H); 1.2 (s, 9H). LCMS (M+H)+ 261.
F. Preparation of N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl- ethoxy)imidazo[2, 1 -b] [ 1 ,3 ]benzothiazol-2-yl]phenyl } urea
1. Example F-l
[00301] The jacket of a 2000-L GL reactor was set to 20 °C and the reactor was charged with 7-(2-morpholin-4-yl-ethoxy)-2-(4-aminophenyl)imidazo[2,l- &]benzothiazole (26.7 kg, 67.8 mol, 1.0 equiv), 3-amino-5-?-butylisoxazole phenyl carbamate (19.4 kg, 74.5 mol, 1.1 equiv), DMAP (0.5 kg, 4.4 mol, 0.06 equiv), and DCM (anhydrous, 260 kg). Agitation was started, triethylamine (1.0 kg, 10.2 mol, 0.15 equiv) was charged followed by additional DCM (5 kg) through the charging line. The reaction was heated to reflux (-40 °C) and agitated for at least 20 hours with complete dissolution observed followed by product crystallizing from solution after -30 minutes. The reaction was sampled and deemed complete when HPLC analysis showed a ratio of compound (VIII) to compound (I) < 1%.
[00302] The reactor was cooled to 0 °C and stirred for at least 2 hours. The content of the reactor were isolated by centrifuge. Each portion was rinsed with 2-3 kg of cold (0 °C) DCM and spun dry for at least 5 minutes with a 10 psi nitrogen purge. For the final portion, the reactor was rinsed with 10 kg of cold (0 °C) DCM and transferred to the centrifuge where it was spun dry for at least 5 minutes with a 10 psi nitrogen purge. The combined filter cakes were transferred to a vacuum tray dryer and dried to constant weight at 50 °C and at least >20 inches of Hg to give the desired product (I) (35.05 kg, 92% yield, HPLC purity = 98.8%). Phenol was the major impurity detected (0.99%); and three other impurities (<0.10%) were detected. JH NMR (300 MHz, DMSO- ) conformed to structure.
2. Example F-2
[00303] A variety of solvents were used in the reaction of Step F to optimize for better yields and purity profiles. The contents of the symmetrical urea impurity (XI) were compared and summarized in the table below:
http://www.google.com/patents/WO2011056939A1?cl=en SE THIS FOR DELETED CLIPS
4. Example F-4
[00305] To Reactor A under a nitrogen atmosphere was added 7-(2-morpholin-4-yl- ethoxy)-2-(4-aminophenyl)imidazo[2,l-&]benzothiazole (1 kg) and DMAP (0.02 kg). To Reactor B under a nitrogen atmosphere was added 3-amino-5-?-butylisoxazole phenyl carbamate (0.73 kg) and DCM (5.6 L). The DCM used had a water content of less than 0.05 % w/w. The mixture in Reactor B was stirred until dissolution. The solution was transferred into Reactor A (the solution can be filtered into Reactor A to remove any insoluble impurities in the carbamate starting material), and the mixture was stirred in Reactor A. Reactor B was washed with DCM (0.8 L) and the wash was transferred into Reactor A. Reactor A was washed with DCM (0.9 L). To Reactor A was added triethylamine (0.1 L) and the charging vessel and lines were rinsed with DCM (0.1 L) into Reactor A. The mixture was then heated to reflux and stirred at reflux until reaction completion (as determined by HPLC to monitor the consumption of starting material).
[00306] The reaction mixture was cooled (e.g. , over 2 to 3 hours) to a temperature of between -5 °C and 5 °C (e.g. , 0 °C). The mixture was then stirred for 2 to 3 hours at a temperature of between -5 °C and 5 °C (e.g. , 0 °C). The suspension was filtered. The solid was washed with cool DCM (2 X 1.5 L) (pre-cooled to a temperature of between -5 °C and 5 °C). The solid was dried under vacuum at a temperature of less than 45 °C until the DCM content was less than 1000 ppm (e.g., below 600 ppm) as analyzed by GC. The desired product was obtained having about 99% purity by HPLC.
G. Preparation of N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl- ethoxy)imidazo[2, l-b] [1 ,3]benzothiazol-2-yl]phenyl }urea dihydrochloride
1. Example G-l
[00307] The jacket of a 2000-L GL reactor was set to 20 °C and the reactor was charged with N-(5-ieri-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo [2, 1-&][1, 3]benzothiazol-2-yl]phenyl}urea (35.0 kg, 62.4 mol, 1.0 equiv) followed by methanol (553 kg). Agitation was started and the reaction mixture was heated to reflux (-65 °C). Concentrated aqueous HC1 (15.4 kg, 156.0 mol, 2.5 equiv) was charged rapidly (<5 minutes) and the charge line was rinsed into the reactor with methanol (12 kg). Addition of less than 2.0 equivalents of HC1 normally resulted in the formation of an insoluble solid. The reaction mixture was heated at reflux for at least 1 hour. Upon HC1 addition, the slurry dissolved and almost immediately the salt started to crystallize, leaving insufficient time for a polish filtration.
[00308] The reactor was cooled to 20 °C and the product was isolated by filtration (Nutsche) rinsing the reactor and then the cake with methanol (58 kg). The solid was then dried in a vacuum oven (50 °C) to constant weight to give the desired
dihydrochloride salt (35 kg, 89% yield, HPLC purity = 99.94%). JH NMR (300 MHz, DMSO-i¾) conformed to structure.
2. Example G-2
[00309] Concentrated HC1 was added to a suspension of N-(5-ieri-butyl-isoxazol-3- yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l-&][l,3]benzothiazol-2- yl]phenyl}urea in warm methanol forming a solution that slowly began to precipitate. The reaction mixture was refluxed for over 2 hours and then stirred overnight at ambient temperature. The dihydrochloride salt was collected and dried under vacuum.
3. Example G-3
[00310] A 50-L 3-neck round bottom flask was equipped with a mechanical agitator, a thermocouple probe, a nitrogen inlet, a drying tube, a reflux condenser, an addition funnel, and a heating mantle. The flask was charged with N-(5-ieri-butyl-isoxazol-3-yl)- N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l-&][l,3]benzothiazol-2-yl]phenyl}urea (775 g, 1.38 mol, 1.0 equiv) and MeOH (40 L, AR). The resulting off-white suspension was heated to reflux (68 °C). A clear solution did not form. HC1 (37% aqueous) (228 mL, 3.46 mol, 2.5 equiv) was added over 5 minutes at 68 °C. The reaction mixture turned into a clear solution and then a new precipitate formed within approximately 3 minutes. Heating was continued at reflux for approximately 5 hours. The reaction mixture was allowed to cool to ambient temperature overnight. The off-white solids were collected by filtration on a polypropylene filter, washing with MeOH (2 X 1 L, AR). [00311] Two lots of material prepared in this manner were combined (740 g and 820 g). The combined solids were slurried in methanol (30 L) over 30 minutes at reflux and allowed to cool to the room temperature. The solids were collected by filtration on a polypropylene filter, rinsing with methanol (2 X 1.5 L). The solids were dried in a vacuum oven (<10 mm Hg) at 40 °C. Yield: 1598 g (84%), off-white solid. HPLC: 98.2% (area). MS: 561.2 (M+l)+. JH NMR (300 MHz, DMSO-i¾) conformed to the structure. Elemental Analysis (EA): Theory, 54.97 %C; 5.41 %H; 13.26 %N; 5.06 %S; 11.19 %C1; Actual, 54.45 %C; 5.46 %H; 13.09 %N; 4.99 %S; 10.91 %C1.
4. Example G-4
[00312] Into a 50-L 3-neck round bottom flask equipped with a mechanical stirrer, a heating mantle, a condenser and a nitrogen inlet, were charged N-(5-ieri-butyl-isoxazol- 3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l-&][l,3]benzothiazol-2- yl]phenyl}urea (1052.4 g, 1.877 mol, 1.0 equiv) and methanol (21 L). The reactor was heated and stirred. At an internal temperature of > 50 °C, cone. HC1 (398.63 mL, 4.693 mol, 2.5 equiv) was charged over 5 minutes through an addition funnel. During the addition, the mixture changed from a pale yellow suspension to a white suspension. The internal temperature was 55 °C at the conclusion of the addition. The mixture was heated to reflux for 1 hour, then heating was discontinued and the mixture was allowed to cool to room temperature. The mixture was filtered in two portions, and each filter cake was washed with methanol (2 X 1 L), transferred to trays and dried in a vacuum oven (45 °C) to constant weight. The dried trays were combined to produce 1141.9 g of the salt (96% yield, 99.1 % HPLC purity, 10.9% chloride by titration).
H. Analytical Data
1. N-(5-ieri-butyl-isoxazol-3-yl)-N’-{ 4-[7-(2-morpholin-4-yl- ethoxy)imidazo[2, l-&] [l ,3]benzothiazol-2-yl]phenyl}urea
dihydrochloride
[00314] A batch of about 30 grams of N-(5-ieri-butyl-isoxazol-3-yl)-N’- {4-[7-(2- morpholin-4-yl-ethoxy)imidazo[2, l-&] [l ,3]benzothiazol-2-yl]phenyl}urea
dihydrochloride was prepared using the methods described herein. This lot was
prepared in accordance with the requirements for production of clinical Active
Pharmaceutical Ingredients (APIs) under GMP conditions. The analytical data for this batch was obtained, and representative data were provided herein. [00315] Summary of analytical data for the dihydrochloride salt.
………………………
EXAMPLE 1. SYNTHESIS OF N-(5-TERT-BUTYL-ISOXAZOL-3-YU- N>-{4-f7-(2-MORPHOLIN-4- YL-ETHOXY)IMID AZO[2,1- BlH,31BENZOTHIAZOL-2-YL|PHENYLiUREA (“COMPOUND Bl”)
[00357] A. The intermediate 2-amino-l,3-benzothiazol-6-ol was prepared according to a slightly modified literature procedure by Lau and Gompf: J. Org. Chem. 1970, 35, 4103- 4108. To a stirred solution of thiourea (7.6 g, 0.10 mol) in a mixture of 200 mL ethanol and 9 mL concentrated hydrochloric acid was added a solution of 1 ,4-benzoquinone (21.6 g, 0.20 mol) in 400 mL of hot ethanol. The reaction was stirred for 24 hours at room temperature and then concentrated to dryness. The residue was triturated with hot acetonitrile and the resulting solid was filtered and dried.
[00358] The free base was obtained by dissolving the hydrochloride salt in water, neutralizing with sodium acetate, and collecting the solid by filtration. The product (2- amino-l,3-benzothiazol-6-ol) was obtained as a dark solid that was pure by LCMS (M+H = 167) and NMR. Yield: 13.0 g (78 %). NMR (DMSO-^) Sl.6 (m, 2H), 6.6 (d, IH). [00359] B. To prepare the 2-(4-nitrophenyl)imidazo[2,l-b][l,3]benzothiazol-7-ol intermediate, 2-amino-l,3-benzothiazol-6-ol (20.0 g, 0.12 mol) and 2-bromo-4′- nitroacetophenone (29.3 g, 0.12 mol) were dissolved in 600 mL ethanol and heated to reflux overnight. The solution was then cooled to O0C in an ice-water bath and the product was collected by vacuum filtration. After drying under vacuum with P2O5 , the intermediate (2- (4-nitrophenyl)imidazo[2,l-£][l,3]benzothiazol-7-ol) was isolated as a yellow solid. Yield: 17.0 g (46 %) NMR (DMSO-(I6) δ 10 (s, IH), 8.9 (s, IH), 8.3 (d, 2H), 8.1 (d, 2H), 7.8 (d, IH), 7.4 (s, IH), 6.9 (d, IH).
[00360] C. To make the 7-(2-morpholin-4-yl-ethoxy)-2-(4-nitro-phenyl)imidazo[2,l-
6][l,3]benzothiazole intermediate: 2-(4-nitrophenyl)imidazo[2,l-6][l,3]benzothiazol-7-ol,
NYI-4144519vl 84 (3.00 g, 9.6 mmol) was suspended in 100 mL dry DMF. To this mixture was added potassium carbonate (4.15 g, 30 mmol, 3 eq), chloroethyl morpholine hydrochloride (4.65 g, 25 mmol, 2.5 eq) and optionally tetrabutyl ammonium iodide (7.39 g, 2 mmol). The suspension was then heated to 900C for 5 hours or until complete by LCMS. The mixture was cooled to room temperature, poured into 800 mL water, and allowed to stand for 1 hour. The resulting precipitate was collected by vacuum filtration and dried under vacuum. The intermediate, (7-(2-morpholin-4-yl-ethoxy)-2-(4-nitro-phenyl)imidazo[2, 1 – b][\, 3]benzothiazole) was carried on without further purification. Yield: 3.87 g (95 %) NMR (DMSO-d6) δ 8.97 (s, IH), 8.30 (d, 2H), 8.0 (d, 2H), 7.9 (d, IH), 7.7 (s, IH), 7.2 (d, IH), 4.1 (t, 2H), 5.6 (m, 4H), 2.7 (t, 2H).
[00361] D. To make the intermediate 7-(2-morpholin-4-yl-ethoxy)-2-(4-amino- phenyl)imidazo[2,l-b][l,3]benzothiazole: To a suspension of 7-(2-morpholin-4-yl-ethoxy)- 2-(4-nitro-phenyl)imidazo[2,l -b][\ , 3]benzothiazole (3.87g, 9.1 mmol) in 100 mL isopropyl alcohol/water (3:1) was added ammonium chloride (2.00 g, 36.4 mmol) and iron powder (5.04 g, 90.1 mmol). The suspension was heated to reflux overnight with vigorous stirring, completion of the reaction was confirmed by LCMS. The mixture was filtered through Celite, and the filtercake was washed with hot isopropyl alcohol (150 mL). The filtrate was concentrated to approximately 1/3 of the original volume, poured into saturated sodium bicarbonate, and extracted 3 times with dichloromethane. The combined organic phases were dried over MgSO4 and concentrated to give the product as an orange solid containing a small amount (4-6 %) of starting material. (Yield: 2.75 g 54 %). 80% ethanol/water may be used in the place of isopropyl alcohol /water – in which case the reaction is virtually complete after 3.5 hours and only traces of starting material are observed in the product obtained. NMR (DMSO-Λfc) δ 8.4 (s, IH), 7.8 (d, IH), 7.65 (d, IH), 7.5 (d, 2H), 7.1 (d, IH), 6.6 (d, 2H), 4.1 (t, 2H), 3.6 (m, 4H), 2.7 (t, 2H).
[00362] E. A suspension of 7-(2-morpholin-4-yl-ethoxy)-2-(4-amino- phenyl)imidazo[2,l-b][l,3]benzothiazole (4.06 g, 10.3 mmol) and 5-tert-butylisoxazole-3- isocyanate (1.994 g, 12 mmol) in toluene was heated at 120 0C overnight. The reaction was quenched by pouring into a mixture of methylene chloride and water containing a little methanol and neutralized with saturated aqueous NaHCO3 solution. The aqueous phase was extracted twice with methylene chloride, the combined organic extracts were dried over
NYI-4144519vl 85 MgSO4 and filtered. The filtrate was concentrated to about 20 ml volume and ethyl ether was added resulting in the formation of a solid. The precipitate was collected by filtration, washed with ethyl ether, and dried under vacuum to give the free base of Compound B 1. Yield: 2.342 g (41 %) NMR (DMSO-J6) £9.6 (br, IH), 8.9 (br, IH), 8.61 (s, IH), 7.86 (d, IH), 7.76 (d, 2H), 7.69 (d, IH), 7.51 (d, 2H), 7.18 (dd, IH), 6.52 (s, IH), 4.16 (t, 2H), 3.59 (t, 4H), 3.36 (overlapping, 4H), 2.72 (t, 2H), 1.30 (s, 9H). NMR (CDCl3) £9.3 (br, IH), 7.84 (m, 4H), 7.59 (d, 2H), 7.49 (d, IH), 7.22 (d, IH), 7.03 (dd, IH), 5.88 (s, IH), 4.16 (t, 2H), 3.76 (t, 4H), 2.84 (t, 2H), 2.61 (t, 4H), 1.37 (s, 9H).
6.2 EXAMPLE 2. ALTERNATIVE SYNTHESIS QF N-(5-TERT-BUTYL- ISOXAZQL-3- YL)-N -{4-[7-q-MORPHOLIN-4- YL- ETHOXYUMID AZOf2,l-BUl,31BENZOTHIAZOL-2- YLIPHENYLIUREA (“COMPOUND Bl”)
[00363] A. To a suspension of the intermediate 2-(4-Nitrophenyl)imidazo[2,l- b][l,3]benzothiazol-7-ol from Example IB (2.24 g, 7.2 mmol) in ethanol (40 mL) was added SnCl2 1H2O (7.9Og, 35 mmol) and heated to reflux. Concentrated HCl was added to the reaction mixture and the precipitate formed gradually. The reaction mixture was heated to reflux for 20 hours and then allowed to cool to room temperature. The solution was poured into ice and neutralized with 10% NaOH and adjusted to approximately pH 6. The organic phase was extracted three times with ethyl acetate (80 mL x 3). Extracts were dried over MgSθ4 and concentrated to give a yellow solid. (1.621 g, 80%). The solid was recrystallized from methanol to give a pure product (1.355 g, 67%).
[00364] B. To a suspension of the intermediate from Step 2A (0.563 g, 2 mmol) in toluene (30 mL) was added 5-tert-butylisoxazole-3-isocyanate (0.332g, 2 mmol) and heated to reflux overnight. LC-MS analysis showed presence of the intermediate but no trace of 5- tert-butylisoxazole-3-isocyanate and an additional 0.166 g of the isocyanate was added. The reaction was again heated to reflux overnight. Completion of reaction was verified by LC- MS. The solvent was removed and the resulting mixture was dissolved in methanol which was removed to give the second intermediate as a solid.
[00365] The mixture was dissolved in CH2Cl2 (150 mL) and washed with saturated
NaHCO3. The organic layer was dried over MgSO4, concentrated, and purified by silica gel chromatography three times, first using a methanol/CH2Cl2 gradient, the second time using a
NYI-4144519vl 86 hexane/ethyl acetate gradient followed by a methanol/ethyl acetate gradient, and a third time using a methanol/CH2Cl2 gradient.
[00366] C. To a suspension of the intermediate from Step 2B (0.1 10 g, 0.25 mmol) in
THF (5mL) was added Ph3P (0.079g, 0.3 mmol), diisopropylazodicarboxylate (0.06 Ig, 0.3 mmol) and 4-morpholinoethanol (0.039 g, 0.3 mmol). The reaction mixture was stirred at room temperature overnight. Completion of the reaction was verified by LC-MS. The solvent was removed and the final product was purified using silica gel chromatography, with methanol in CH2Cl2 (0.030g, 21%).
6.3 EXAMPLE 3. BULK SYNTHESIS OF N-(5-TERT-BUTYL- ISOXAZOL-3-YL)-N’-f4-[7-(2-MORPHOLIN-4-YL- ETHOXY^IMID AZO[2α-BUlJlBENZOTHIAZOL-2- YLlPHENYLiUREA (“COMPOUND Bl”)
[00367] A multi-step reaction scheme that was used to prepare bulk quantities of
Compound Bl is depicted in FIG. 66a and FIG. 66b, and is described further below. [00368] Step 1 : Preparation of 2- Amino-6-hydroxybenzothiazole (Intermediate 1). 2-
Amino-6-methoxybenzothiazole is reacted with hot aqueous HBr for about 3 hrs and then the clear solution is cooled to ambient temperature overnight. The precipitated solids are collected, dissolved in hot water and the pH is adjusted to between 4.5-5.5. The resultant solids are collected, dried and recrystallized from Isopropanol. Second crop material is collected. The solids are vacuum dried to give Intermediate 1.
[00369] Step 2: Preparation of 2-(4-Nitrophenyl) imidazo [2J-b]benzothiazol-7-ol
(Intermediate 2). 2-Amino-6-hydroxybenzothiazole, 2-Bromo-4-nitroacetophenone and absolute Ethanol are added together and heated to reflux for approximately 24 hours. Tetrabutylammonium iodide is added and the reaction is refluxed an additional 12 hours. The resulting yellow suspension is cooled and the solids collected and washed with Ethanol and Diethyl ether. The solids are dried under vacuum to give Intermediate 2. [00370] Step 3: Preparation of 7-(2-Morpholin-4-yl-ethoxy)-2-(4-nitrophenyl) imidazo
[2,1-b] benzothiazole (Intermediate 3). Intermediate 2, 4-(2-Chloroethyl)morpholine hydrochloride, Potassium carbonate and Tetrabutylammonium iodide are added to N,N- Dimethylformamide forming a yellow suspension that is heated for over 3 hours. The reaction is cooled and the solids are collected, slurried into water, filtered, slurried into
NYl-4 l4451′)v l 87 acetone, filtered and washed with Acetone to give yellow solids that are dried under vacuum to give Intermediate 3.
[O0371] Step 4: Preparation of 7-(2-Moφholin-4-yl-ethoxy)-2-(4-aminophenyl) imidazo [2,1 -b] benzothiazole (Intermediate 4). Intermediate 3 is dissolved into Methanol and THF and placed in a Hydrogenator. Raney Nickel is added and the vessel is pressurized with Hydrogen and stirred for >24 hrs. The reaction mixture is concentrated to a thick paste and diluted with Methyl tert-butyl ether. The resulting solids are filtered and washed with Methyl tert-butyl ether and dried under vacuum to give Intermediate 4. [O0372] Step 5: Preparation of {[5-(tert-Butyl) isoxazol-3-vnatnino}-N-{4-r7-(2- morpholin-4-yl-ethoxy)(4-hvdroimidazolo[2J-blbenzothiazol-2-yl)]phenyl|carboxamide (Compound Bl). 3 -Amino- 5 -tert-butyl isoxazole in Methylene chloride is added to a vessel containing toluene which is cooled to approx 0 0C. Triphosgene is then added and the reaction mixture is cooled to below -15 0C. Triethylamine is added, followed by Intermediate 4. The mixture is heated to distill off the Methylene chloride and then heated to over 60 0C for over 12 hours and cooled to 50-60 °C. The resulting solids are filtered, washed with Heptane, slurried with 4% sodium hydroxide solution, and filtered. The solids are then washed with Methyl tert-butyl ether followed by Acetone and dried under vacuum to give Compound Bl.
6.4 EXAMPLE 4. EXAMPLES OF PREPARATION OF COMPOUND Bl HCL SALT
[00373] Example A: For the preparation of a hydrochloride salt of Compound Bl5 N-
(5-tert-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,l- b][l,3]benzothiazol-2-yl]phenyl}urea hydrochloride, the free base was dissolved in a mixture of 20 ml methylene chloride and 1 ml methanol. A solution of 1.0 M HCl in ethyl ether (1.1 eq.) was added dropwise, followed by addition of ethyl ether. The precipitate was collected by filtration or centrirugation and washed with ethyl ether to give a hydrochloride salt of Compound Bl. Yield: 2.44 g (98 %) NMR (DMSO-^) S X 1.0 (br, IH), 9.68 (s, IH), 9.26 (s, IH), 8.66 (s, IH), 7.93 (d, IH), 7.78 (m, 3H), 7.53 (d, 2H), 7.26 (dd, IH), 6.53 (s, IH), 4.50 (t, 2H), 3.97 (m, 2H), 3.81 (t, 2H), 3.6 (overlapping, 4H), 3.23 (m, 2H), 1.30 (s, 9H). [00374] Example B: Concentrated HCl is added to a suspension of Compound Bl in warm methanol forming a solution that slowly begins to precipitate. The reaction mixture is
NYI-4144519vl 88 refluxed for over 2 hrs and then stirred overnight at ambient temperature. The HCl salt is collected and dried under vacuum.
[00375] Example C: Materials: {[5-(tert-Butyl) isoxazol-3-yl]amino}-N-{4-[7-(2- morpholin-4-yl-ethoxy)(4-hydroimidazolo[2,l-6]benzothiazol-2-yl)] phenyl }carboxamide (775 g, 1.38 mol, 1.0 eq); HCl 37% aqueous (288 mL, 3.46 mol, 2.5 eq); Methanol (MeOH, AR) (40L). Procedure: (Step 1) Equipped a 5OL 3-neck round bottom flask with a mechanical agitator, thermocouple probe, Nitrogen inlet, drying tube, reflux condenser, addition funnel and in a heating mantle. (Step 2) Charged the flask with {[5-(tert-Butyl) isoxazol-3-yl] amino}-N-{4-[7-(2-morpholin-4-yl-ethoxy)(4-hydroimidazolo[2,l- b]benzothiazol-2-yl)] phenyl jcarboxamide (775g) and MeOH, AR (40L). Heat the resulting off-white suspension to reflux (680C). A clear solution did not form. (Step 3) Added HCl (37% aqueous) (228 mL) over 5 minutes at 68°C. The reaction mixture turned into a clear solution and then a new precipitate formed within approximately 3 minutes. Continued heating at reflux for approximately 5 hours. Allowed the reaction mixture to cool to ambient temperature overnight. (Step 4) Collected the off-white solids by filtration onto a polypropylene filter, washing the solids with MeOH, AR (2 x 1 L). (Step 5) Combined two lots of material prepared in this manner (74Og and 82Og). Slurried the combined solids in Methanol (30 L) over 30 minutes at reflux and cool to the room temperature. (Step 6) Collected the solids by filtration onto a polypropylene filter, rinsing with Methanol (2 x 1.5L). (Step 7) Dried the solids in a vacuum oven (<10mniHg) at 400C. Yield: 1598 g (84%), off-white solid; HPLC: 98.2% (area); MS: 561.2 (M+l); IH NMR: conforms (300 MHz, DMSO-d6); Elemental Analysis (EA): Theory = 54.97 %C; 5.41 %H; 13.26 %N; 5.06 %S; 11.19 %C1; Actual = 54.45 %C; 5.46 %H; 13.09 %N; 4.99 %S; 10.91 %C1.
NYl-4I44519v! 89 [00376] Examples of Compound Bl HCl salt synthesis
[00377] Example D: In a 50-L 3-neck round bottom flask equipped with a mechanical stirrer, heating mantle, condenser and nitrogen inlet was charged Compound Bl (1052.4 g, 1.877 mol, 1.00 equiv.) and methanol (21 L). The reactor was heated and stirred. At an internal temperature > 50 0C, cone. HCl (398.63 mL, 4.693 mol, 2.5 equiv.) was charged over 5 minutes through an addition funnel. With the addition, the reaction changed from a pale yellow suspension to a white suspension. The internal temperature was 55 0C at the conclusion of the addition. The reaction was heated to reflux for 1 hour, then heating discontinued and the reaction allowed to cool to room temperature. The reaction was filtered in two portions, each filter cake washed with methanol (2 x 1 L), transferred to trays and dried in a vacuum oven (45 0C) to constant weight. The dried trays were combined to produce 1141.9 g, 96% yield, 99.1 % HPLC purity, 10.9% chloride by titration.
Solid Forms Comprising the HCl Salt of Compound Bl 6.6.2.1 Preparation of Solid Forms
6.6.2.2 Cold Precipitation Experiments
NYl-4144519vl 102 6.6.2.3 Slurry Experiments
NYI-41445 l9vl 103 6.6.2.4 Additional Preparation of Solid Forms Comprising the HCI Salt of Compound Bl
NYl-4144519v l 104
NYM 144519vl 105
N Y l -4 1 4 4 5 1 9 v l 1 0 6
NYI-4I44519vi 107
N V I 4 1 4 4 5 1 9 1 0 8
“Abbreviations in Table: CC = crash cool, CP = crash precipitation, EtOAc = ethyl acetate, FE = fast evaporation, VD = vapor diffusion, IPA = isopropanol, MEK = methyl ethyl ketone (2-butanone), RE = rotary evaporation, RT = room (ambient) temperature, SC = slow cool, SE = slow evaporation, THF = tetrahydrofuran, TFE = 2,2,2=trifluoroethanol.
6.6.2.5 Scale-up Experiments of Involving Crystal Forms Comprising the HCl Salt of Compound Bl
NYI-4144519v l 109
Abbreviations in Table: CC = crash cool, CP = crash precipitation, EtOAc = ethyl acetate, FE = fast evaporation, IPA = isopropanol, MEK = methyl ethyl ketone (2-butanone), RE = rotary evaporation, RT = room (ambient) temperature, SC = slow cool, SE = slow evaporation, THF = tetrahydrofuran, TFE = 2,2,2=trifluoroethanol.
……………………
Identification of N-(5-tert-butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,1-b][1,3]benzothiazol-2-yl]phenyl}urea dihydrochloride (AC220), a uniquely potent, selective, and efficacious FMS-like tyrosine kinase-3 (FLT3) inhibitor
J Med Chem 2009, 52(23): 7808
http://pubs.acs.org/doi/full/10.1021/jm9007533
General Procedure E for Preparation of Hydrochloride Salt
References
- Chao, Qi; Sprankle, Kelly G.; Grotzfeld, Robert M.; Lai, Andiliy G.; Carter, Todd A.; Velasco, Anne Marie; Gunawardane, Ruwanthi N.; Cramer, Merryl D.; Gardner, Michael F.; James, Joyce; Zarrinkar, Patrick P.; Patel, Hitesh K.; Bhagwat, Shripad S. (2009). “Identification of N-(5-tert-Butyl-isoxazol-3-yl)-N’-{4-[7-(2-morpholin-4-yl-ethoxy)imidazo[2,1-b][1,3]benzothiazol-2-yl]phenyl}urea Dihydrochloride (AC220), a Uniquely Potent, Selective, and Efficacious FMS-Like Tyrosine Kinase-3 (FLT3) Inhibitor”. Journal of Medicinal Chemistry 52 (23): 7808–7816.
- Drug Tames Refractory AML. ASH Dec 2012
- NMR……….http://file.selleckchem.com/downloads/nmr/S152601-AC-220-HNMR-Selleck.pdf
- HPLC………http://file.selleckchem.com/downloads/hplc/S152601-AC-220-HPLC-Selleck.pdf
HS 10340
HS-10340
CAS 2156639-66-4
CAS 2307670-65-9
Jiangsu Hansoh Pharmaceutical Group Co Ltd
Being investigated by Jiangsu Hansoh, Shanghai Hansoh Biomedical and Changzhou Hengbang Pharmaceutical ; in June 2018, the product was being developed as a class 1 chemical drug in China.
Useful for treating liver cancer, gastric cancer and prostate cancer.
Use for treating cancers, liver cancer, gastric cancer, prostate cancer, skin cancer, ovary cancer, lung cancer, breast cancer, colon cancer, glioma and rhabdomyosarcoma
PATENT
WO2017198149
where it is claimed to be an FGFR-4 inhibitor for treating liver and prostate cancers, assigned to Jiangsu Hansoh Pharmaceutical Group Co Ltd and Shanghai Hansoh Biomedical Co Ltd .
PATENT
WO2019085860
PATENT
WO-2019085927
https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2019085927&tab=FULLTEXT
Novel crystalline salt (such as hydrochloride, sulfate, methane sulfonate, mesylate, besylate, ethanesulfonate, oxalate, maleate, p-toluenesulfonate) forms of FGFR4 inhibitor, particularly N-[5-cyano-4-[[(1R)-2-methoxy-1-methyl-ethyl]amino]-2-pyridyl]-7-formyl-6-[(2-oxo-1,3-oxazepan-3-yl)methyl]-3,4-dihydro-2H-1,8-naphthyridine-1-carboxamide (designated as Forms I- IX), compositions comprising them and their use as an FGFR4 inhibitor for the treatment of cancer such as liver cancer, gastric cancer, prostate cancer, skin cancer, ovarian cancer, lung cancer, breast cancer, colon cancer and glioma or rhabdomyosarcoma are claimed.
///////////HS-10340 , HS 10340 , HS10340, CANCER, Jiangsu Hansoh, Shanghai Hansoh Biomedical, Changzhou Hengbang, CHINA, liver cancer, gastric cancer, prostate cancer, skin cancer, ovary cancer, lung cancer, breast cancer, colon cancer, glioma, rhabdomyosarcoma
C[C@H](COC)Nc1cc(ncc1C#N)NC(=O)N4CCCc3cc(CN2CCCCOC2=O)c(C=O)nc34
CCS(=O)(=O)O.C[C@H](COC)Nc1cc(ncc1C#N)NC(=O)N4CCCc3cc(CN2CCCCOC2=O)c(C=O)nc34
FDA approves first treatment Soliris (eculizumab) for neuromyelitis optica spectrum disorder, a rare autoimmune disease of the central nervous system
The U.S. Food and Drug Administration today approved Soliris (eculizumab) injection for intravenous use for the treatment of neuromyelitis optica spectrum disorder (NMOSD) in adult patients who are anti-aquaporin-4 (AQP4) antibody positive. NMOSD is an autoimmune disease of the central nervous system that mainly affects the optic nerves and spinal cord.
“Soliris provides the first FDA-approved treatment for neuromyelitis optica spectrum disorder, a debilitating disease that profoundly impacts patients’ lives,” said Billy Dunn, M.D., director of the Division of Neurology Products in the FDA’s Center for Drug Evaluation and Research. “This approval changes the landscape of therapy for patients with NMOSD. Having an approved therapy for this condition is the culmination of extensive work we have engaged in with drug companies to …
- June 27, 2019
The U.S. Food and Drug Administration today approved Soliris (eculizumab) injection for intravenous use for the treatment of neuromyelitis optica spectrum disorder (NMOSD) in adult patients who are anti-aquaporin-4 (AQP4) antibody positive. NMOSD is an autoimmune disease of the central nervous system that mainly affects the optic nerves and spinal cord.
“Soliris provides the first FDA-approved treatment for neuromyelitis optica spectrum disorder, a debilitating disease that profoundly impacts patients’ lives,” said Billy Dunn, M.D., director of the Division of Neurology Products in the FDA’s Center for Drug Evaluation and Research. “This approval changes the landscape of therapy for patients with NMOSD. Having an approved therapy for this condition is the culmination of extensive work we have engaged in with drug companies to expedite the development and approval of safe and effective treatments for patients with NMOSD, and we remain committed to these efforts for other rare diseases.”
In patients with NMOSD, the body’s immune system mistakenly attacks healthy cells and proteins in the body, most often in the optic nerves and spinal cord. Individuals with NMOSD typically have attacks of optic neuritis, which causes eye pain and vision loss. Individuals also can have attacks resulting in transverse myelitis, which often causes numbness, weakness, or paralysis of the arms and legs, along with loss of bladder and bowel control. Most attacks occur in clusters, days to months to years apart, followed by partial recovery during periods of remission. Approximately 50% of patients with NMOSD have permanent visual impairment and paralysis caused by NMOSD attacks. According to the National Institutes of Health, women are more often affected by NMOSD than men and African Americans are at greater risk of the disease than Caucasians. Estimates vary, but NMOSD is thought to impact approximately 4,000 to 8,000 patients in the United States.
NMOSD can be associated with antibodies that bind to a protein called aquaporin-4 (AQP4). Binding of the anti-AQP4 antibody appears to activate other components of the immune system, causing inflammation and damage to the central nervous system.
The effectiveness of Soliris for the treatment of NMOSD was demonstrated in a clinical study of 143 patients with NMOSD who had antibodies against AQP4 (anti-AQP4 positive) who were randomized to receive either Soliris treatment or placebo. Compared to treatment with placebo, the study showed that treatment with Soliris reduced the number of NMOSD relapses by 94 percent over the 48-week course of the trial. Soliris also reduced the need for hospitalizations and the need for treatment of acute attacks with corticosteroids and plasma exchange.
Soliris has a boxed warning to alert health care professionals and patients that life-threatening and fatal meningococcal infections have occurred in patients treated with Soliris, and that such infections may become rapidly life-threatening or fatal if not recognized and treated early. Patients should be monitored for early signs of meningococcal infections and evaluated immediately if infection is suspected. Use should be discontinued in patients who are being treated for serious meningococcal infections. Health care professionals should use caution when administering Soliris to patients with any other infection. In the NMOSD clinical trial, no cases of meningococcal infection were observed.
Soliris is available only through a restricted program under a Risk Evaluation and Mitigation Strategy (REMS). Prescribers must enroll in the REMS program. Prescribers must counsel patients about the risk of meningococcal infection, provide the patients with the REMS educational materials and ensure patients are vaccinated with meningococcal vaccine(s). The drug must be dispensed with the FDA-approved patient Medication Guide that provides important information about the drug’s uses and risks.
The most frequently reported adverse reactions reported by patients in the NMOSD clinical trial were: upper respiratory infection, common cold (nasopharyngitis), diarrhea, back pain, dizziness, influenza, joint pain (arthralgia), sore throat (pharyngitis) and contusion.
The FDA granted the approval of Soliris to Alexion Pharmaceuticals.
Soliris was first approved by the FDA in 2007. The drug is approved to reduce destruction of red blood cells in adults with a rare blood disease called paroxysmal nocturnal hemoglobinuria, for the treatment of adults and children with a rare disease that causes abnormal blood clots to form in small blood vessels in the kidneys (atypical hemolytic uremic syndrome to inhibit complement-mediated thrombotic microangiopathy), and for the treatment of adults with Myasthenia Gravis who are anti-acetylcholine receptor antibody positive.
The FDA granted this application Priority Review. The use for NMOSD received Orphan Drug designation, which provides incentives to assist and encourage the development of drugs for rare diseases.
///////////////fda 2019, Soliris, eculizumab, neuromyelitis optica spectrum disorder, Orphan Drug, Priority Review
CS 3001
CS-3001
BB 7, VX 033
- Molecular Weight, 478.37
C17 H18 Br F2 N3 O2 S2
CStone Pharmaceuticals Co Ltd, JUNE 2018 IND FILED CHINA
URAT1 inhibitor – useful for treating hyperuricemia and gout.
The compound was originally claimed in WO2017202291 , covering thiophene derivative URAT1 inhibitors, useful for treating hyperuricemia and gouty arthritis, assigned to Medshine Discovery Inc , but naming the inventors.and has been reported in some instances to be a URAT1 modulator. In June 2018, an IND application was filed in
WO-2019101058
https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2019101058&tab=FULLTEXT&maxRec=1000
Novel crystalline forms of URAT1 inhibitor (designated as Forms A and B) are claimed. The compounds are disclosed to be useful for treating hyperuricemia and gouty arthritis.
Novel crystalline forms of a URAT1 inhibitor, designated as Forms A and B, and their preparation.
For example, remove 2.0 mL of phosphoric acid into 2000 mL of water, sonicate for 10 minutes, mix, and let cool to room temperature as mobile phase A.
////////////CS-3001, BB 7, VX 033, CHINA, PRECLINICAL, CStone Pharmaceuticals, URAT1 inhibitor, hyperuricemia, gout
O=C(O)C(C)(C)Sc4nnc(Br)n4c2sc(c1CC(F)(F)CCc12)C3CC3
DR ANTHONY MELVIN CRASTO
ORGANIC SPECTROSCOPY
New Drug Approvals 