Abiraterone acetate

• Molecular FormulaC₂₆H₃₃NO₂
• Average mass391.546 Da

Abiraterone, CB-7598, アビラテロン酢酸エステル

Abiraterone is a derivative of steroidal progesterone and is an innovative drug that offers clinical benefit to patients with hormone refractory prostate cancer. Abiraterone is administered as an acetate salt prodrug because it has a higher bioavailability and less susceptible to hydrolysis than abiraterone itself. FDA approved on April 28, 2011.

Used in combination with prednisone for the treatment of metastatic, castration-resistant prostate cancer.

• Originator The Institute of Cancer Research
• Developer All Ireland Cooperative Oncology Research Group; Cancer Research UK; Cougar Biotechnology; Janssen Research & Development; Johnson & Johnson; UNICANCER
• Class Androstenols; Antiandrogens; Antineoplastics; Small molecules
• Mechanism of Action CYP17A1 protein inhibitors

Highest Development Phases

• Marketed Prostate cancer
• Phase II Breast cancer; Ovarian cancer
• No development reported Congenital adrenal hyperplasia

Most Recent Events

• 06 Jun 2018 The National Institute for Health and Clinical Excellence does not recommend abiraterone for Prostate cancer (Combination therapy, First-line therapy, Hormone refractory, Metastatic disease)
• 06 Mar 2018 Janssen initiates a phase II trial for Prostate cancer (Combination therapy, Hormone refractory, Metastatic disease, Second-line therapy or greater) in USA (PO) (NCT03360721)
• 01 Mar 2018 Janssen plans the phase II OPTIMABI trial in Prostate cancer (Hormone refractory, Metastatic disease) in France (PO, Tablet) (NCT03458247)
• Abiraterone is associated with decreases in PSA levels, tumor shrinkage (as evaluated by RECIST criteria), radiographic regression of bone metastases and improvement in pain. Levels of adrenocorticotropic hormones increased up to 6-fold but this can be suppressed by dexamethasone.

FDA

NDA 202379, ZYTIGA (abiaterone acetate)

(3β)-17-(3-pyridinyl)androsta-5,16-dien-3-yl acetate

OND Division: NDA: Applicant: Stamp Date: PDUFA Goal Date: Established Name: Trade Name Dosage Form and Strength: Route of Administration: Indication: eCTD Reference for CMC Regulatory Filing Related IND Assessed by: Division of Drug Oncology Products 202-379 Centocor Ortho Biotech, Inc. 20 December, 2010 20 June, 2011 (Priority) Abiraterone Acetate ZYTIGA (proposed) Tablet – 250 mg Oral Indicated with prednisone for the treatment of metastatic (castrationresistant prostate cancer) in patients who have received prior chemotherapy containing a

https://www.accessdata.fda.gov/drugsatfda_docs/nda/2011/202379Orig1s000ChemR.pdf

Abiraterone acetate, the drug substance, is an acetyl ester of abiraterone. It is a pro-drug of the active metabolite abiraterone. Abiraterone acetate is converted in vivo to abiraterone which selectively inhibits the enzyme CYP17. Abiraterone acetate is designated chemically as (3β)-17- (3-pyridinyl)androsta-5,16-dien-3-yl acetate. It is a white to off-white, non-hygroscpic, crystalline powder. It is freely soluble in organic solvents like tetrahydrofuran and dichloromethane but practically insoluble in water. It shows some solubility in 0.1N HCl. It should be noted that abiraterone acetate contains a . The dissociation constant (pKa) of abiraterone acetate is 5.19. It indicates that most of the abiraterone acetate will be soluble in stomach pH and most of the drug will be absorbed in the unionized form in the intestine at higher pH. The partition coefficient (log P) value of abiraterone acetate is 5.12 indicating high lipophilicity. Based on low aqueous solubility and low permeability thru the cells in GI tract, the drug substance is considered BCS Class IV.

Abiraterone acetate, sold under the brand name Zytiga among others, is an antiandrogen medication which is used in the treatment of prostate cancer.^[1] It is specifically indicated for use in conjunction with castration and prednisone for the treatment of metastaticcastration-resistant prostate cancer (mCRPC) and in the treatment of metastatic high-risk castration-sensitive prostate cancer (mCSPC).^[1] It is taken by mouth once per day with food.^[1]
Side effects of abiraterone acetate include fatigue, arthralgia, hypertension, nausea, edema, hypokalemia, hot flashes, diarrhea, vomiting, cough, headache, glucocorticoid deficiency, mineralocorticoid excess, and hepatotoxicity among others.^[1] The drug is an androgen synthesis inhibitor – specifically, a CYP17A1 inhibitor – and thereby inhibits the production of androgens like testosterone and dihydrotestosterone in the body.^[1] In doing so, it prevents the effects of these hormones in the prostate gland and elsewhere in the body.^[1] Abiraterone acetate is a prodrug of abiraterone.^[1]Abiraterone acetate, sold under the brand name Zytiga among others, is an antiandrogen medication which is used in the treatment of prostate cancer.^[1] It is specifically indicated for use in conjunction with castration and prednisone for the treatment of metastaticcastration-resistant prostate cancer (mCRPC) and in the treatment of metastatic high-risk castration-sensitive prostate cancer (mCSPC).^[1] It is taken by mouth once per day with food.^[1]

Abiraterone acetate was first described in 1993 and was introduced for medical use in 2011.^[5][6][7] It was approved for the treatment of mCRPC in 2011 and was subsequently approved for the treatment of mCSPC in 2018.^[8] The medication is marketed widely throughout the world.^[9] It is not available as a generic medication.^[10]

Medical uses

Prostate cancer

Abiraterone acetate is indicated for use in combination with prednisone, a corticosteroid, as a treatment for mCRPC (previously called hormone-resistant or hormone-refractory prostate cancer).^{[11][12][13][14]} This is a form of prostate cancer that is not responding to first-line androgen deprivation therapy or treatment with androgen receptor antagonists. Abiraterone acetate has received FDA (28 April 2011), EMA (23 September 2011), MHRA (5 September 2011) and TGA (1 March 2012) approval for this indication.^{[11][12][13][14]} In Australia it is covered by the Pharmaceutical Benefits Scheme when being used to treat castration-resistant prostate cancer and given in combination with prednisone/prednisolone (subject to the conditions that the patient is not currently receiving chemotherapy, is either resistant or intolerant of docetaxel, has a WHO performance status of <2, and his disease has not since become progressive since treatment with PBS-subsidised abiraterone acetate has commenced).^[15]

Clinical effectiveness

A phase III study in subjects previously treated with docetaxel started in 2008.^[16] In September 2010, an independent panel found that the interim results in patients previously treated with docetaxel were so much better compared to those treated with placebo that it was unethical to keep half the study participants on placebo, and all patients began receiving the drug. Overall survival was increased by 3.9 months in to this study (14.8 months versus 10.9 months for placebo).^[17]

A placebo-controlled double-blind randomized phase III study in patients with castration-refractory prostate cancer but who had not received chemotherapy opened to accrual in April 2009.^[18][19] 1,088 men received either abiraterone acetate (1000 mg daily) plus prednisone (5 mg twice daily), or placebo plus prednisone. The median radiographic progression-free survival was 16.5 months with abiraterone acetate–prednisone and 8.3 months with prednisone alone (hazard ratio (HR) = 0.53; 95% confidence interval (CI), 0.45 to 0.62; P<0.001). After a median follow-up period of 22.2 months, overall survival was better with abiraterone acetate plus prednisone (median not reached) compared to placebo plus prednisone (27.2 months); HR = 0.75; 95% CI, 0.61 to 0.93; P=0.01).^[20]

Available forms

Abiraterone acetate is available in the form of 250 mg and 500 mg film-coated oral tablets and 250 mg uncoated oral tablets.^[1] It is used at a dosage of 1,000 mg orally once per day with food in conjunction with castration (via GnRH analogue therapy or orchiectomy) and in combination with 5 mg prednisone orally twice per day.^[1]

Contraindications

Contraindications include hypersensitivity to abiraterone acetate. Although documents state that it should not be taken by women who are or who may become pregnant,^[12][21] there is no medical reason that any woman should take it. Women who are pregnant should not even touch the pills unless they are wearing gloves.^[21] Other cautions include severe baseline hepatic impairment, mineralocorticoid excess, cardiovascular disease including heart failure and hypertension, uncorrected hypokalemia, and adrenocorticoid insufficiency.^[22]

Side effects

Side effects by frequency:^{[11][12][13][14][22]}

Very common (>10% frequency):

Common (1-10% frequency):

Uncommon (0.1-1% frequency):

• Adrenal insufficiency
• Myopathy
• Rhabdomyolysis

Rare (<0.1% frequency):

• Allergic alveolitis

Overdose

Clinical experience with overdose of abiraterone acetate is limited.^[1] There is no specific antidote for abiraterone acetate overdose, and treatment should consist of general supportive measures, including monitoring of cardiac and liver function.^[1]

Interactions

Abiraterone acetate is a CYP3A4 substrate and hence should not be administered concurrently with strong CYP3A4 inhibitors such as ketoconazole, itraconazole, clarithromycin, atazanavir, nefazodone, saquinavir, telithromycin, ritonavir, indinavir, nelfinavir, voriconazole) or inducers such as phenytoin, carbamazepine, rifampin, rifabutin, rifapentine, phenobarbital.^[22][21] It also inhibits CYP1A2, CYP2C9, and CYP3A4 and likewise should not be taken concurrently with substrates of any of these enzymes that have a narrow therapeutic index.^[22][21]

Pharmacology

Pharmacodynamics

Antiandrogenic activity

Abiraterone, the active metabolite of abiraterone acetate, inhibits CYP17A1, which manifests as two enzymes, 17α-hydroxylase (IC₅₀ = 2.5 nM) and 17,20-lyase (IC₅₀ = 15 nM) (approximately 6-fold more selective for inhibition of 17α-hydroxylase over 17,20-lyase)^[23][24] that are expressed in testicular, adrenal, and prostatic tumor tissues. CYP17A1 catalyzes two sequential reactions: (a) the conversion of pregnenolone and progesterone to their 17α-hydroxy derivatives by its 17α-hydroxylase activity, and (b) the subsequent formation of dehydroepiandrosterone (DHEA) and androstenedione, respectively, by its 17,20-lyase activity.^[25] DHEA and androstenedione are androgens and precursors of testosterone. Inhibition of CYP17A1 activity by abiraterone thus decreases circulating levels of androgens such as DHEA, testosterone, and dihydrotestosterone (DHT). Abiraterone acetate, via its metabolite abiraterone, has the capacity to lower circulating testosterone levels to less than 1 ng/dL (i.e., undetectable) when added to castration.^[23][26] These concentrations are considerably lower than those achieved by castration alone (~20 ng/dL).^[26] The addition of abiraterone acetate to castration was found to reduce levels of DHT by 85%, DHEA by 97 to 98%, and androstenedione by 77 to 78% relative to castration alone.^[26] In accordance with its antiandrogenic action, abiraterone acetate decreases the weights of the prostate gland, seminal vesicles, and testes.^[27]

Abiraterone also acts as a partial antagonist of the androgen receptor (AR), and as an inhibitor of the enzymes 3β-hydroxysteroid dehydrogenase (3β-HSD), CYP11B1 (steroid 11β-hydroxylase), CYP21A2 (Steroid 21-hydroxylase), and other CYP450s (e.g., CYP1A2, CYP2C9, and CYP3A4).^{[22][28][29][30]} In addition to abiraterone itself, part of the activity of the drug has been found to be due to a more potent active metabolite, δ⁴-abiraterone (D4A), which is formed from abiraterone by 3β-HSD.^[31] D4A is an inhibitor of CYP17A1, 3β-hydroxysteroid dehydrogenase/Δ^5-4 isomerase, and 5α-reductase, and has also been found to act as a competitive antagonist of the AR reportedly comparable to the potent antagonist enzalutamide.^[31] However, the initial 5α-reduced metabolite of D4A, 3-keto-5α-abiraterone, is an agonist of the AR, and promotes prostate cancer progression.^[32] Its formation can be blocked by the coadministration of dutasteride, a potent and selective 5α-reductase inhibitor.^[32]

Estrogenic activity

There has been interest in the use of abiraterone acetate for the treatment of breast cancer due to its ability to lower estrogen levels.^[33] However, abiraterone has been found to act as a direct agonist of the estrogen receptor, and induces proliferation of human breast cancer cells in vitro.^[33] If abiraterone acetate is used in the treatment of breast cancer, it should be combined with an estrogen receptor antagonist like fulvestrant.^[33] In spite of its antiandrogenic and estrogenic properties, abiraterone acetate does not appear to produce gynecomastia as a side effect.^[34]

Other activities

Due to inhibition of glucocorticoid biosynthesis, abiraterone acetate can cause glucocorticoid deficiency, mineralocorticoid excess, and associated adverse effects.^[35] This is why the medication is combined with prednisone, a corticosteroid, which serves as a means of glucocorticoid replacement and prevents mineralocorticoid excess.^[36]

Abiraterone acetate, along with galeterone, has been identified as an inhibitor of sulfotransferases (SULT2A1, SULT2B1b, SULT1E1), which are involved in the sulfation of DHEA and other endogenous steroids and compounds, with K_i values in the sub-micrmolar range.^[37]

Pharmacokinetics

After oral administration, abiraterone acetate, the prodrug form in the commercial preparation, is converted into the active form, abiraterone. This conversion is likely to be esterase-mediated and not CYP-mediated. Administration with food increases absorption of the drug and thus has the potential to result in increased and highly variable exposures; the drug should be consumed on an empty stomach at least one hour before or two hours after food. The drug is highly protein bound (>99%), and is metabolised in the liver by CYP3A4 and SULT2A1 to inactive metabolites. The drug is excreted in feces (~88%) and urine (~5%), and has a terminal half-life of 12 ± 5 hours.^[21]

Chemistry

Abiraterone acetate, also known as 17-(3-pyridinyl)androsta-5,16-dien-3β-ol acetate, is a synthetic androstane steroid and a derivative of androstadienol (androsta-5,16-dien-3β-ol), an endogenous androstane pheromone. It is specifically a derivative of androstadienol with a pyridine ring attached at the C17 position and an acetate ester attached to the C3β hydroxyl group. Abiraterone acetate is the C3β acetate ester of abiraterone.

History

In the early 1990s, Mike Jarman, Elaine Barrie, and Gerry Potter of the Cancer Research UK Centre for Cancer Therapeutics in the Institute of Cancer Research in London set out to develop drug treatments for prostate cancer. With the nonsteroidal androgen synthesis inhibitor ketoconazole as a model, they developed abiraterone, filing a patent in 1993 and publishing the first paper describing it the following year.^[5][38] Rights for commercialization of the drug were assigned to BTG, a UK-based specialist healthcare company. BTG then licensed the product to Cougar Biotechnology, which began development of the commercial product.^[39] In 2009, Cougar was acquired by Johnson & Johnson, which developed and sells the commercial product, and is conducting ongoing clinical trials to expand its clinical uses.^[40]

Abiraterone acetate was approved by the United States Food and Drug Administration on April 28, 2011.^[6][7] The FDA press release made reference to a phase III clinical trial in which abiraterone use was associated with a median survival of 14.8 months versus 10.9 months with placebo; the study was stopped early because of the successful outcome.^[41]Abiraterone acetate was also licensed by the European Medicines Agency.^[42] Until May 2012 the National Institute for Health and Clinical Excellence (NICE) did not recommend use of the drug within the NHS on cost-effectiveness grounds. This position was reversed when the manufacturer submitted revised costs.^[43] The use is currently limited to men who have already received one docetaxel-containing chemotherapy regimen.^[44][45]

Society and culture

Generic names

Abiraterone acetate is the generic name of the drug and its USAN, BANM, and JAN, while abiraterone is the INN and BAN of abiraterone, its deacetylated form.^[9] Abiraterone acetate is also known by its developmental code names CB-7630 and JNJ-212082, while CB-7598 was the developmental code name of abiraterone.^[9][46]

Brand names

Abiraterone acetate is marketed by Janssen Biotech (a subsidiary of Johnson & Johnson) under the brand name Zytiga.^[9] In addition, Intas Pharmaceuticals markets the drug under the brand name Abiratas, Cadila Pharmaceuticals markets the drug as Abretone, and Glenmark Pharmaceuticals as Abirapro.^{[citation needed]}

Availability

Abiraterone acetate is marketed widely throughout the world, including in the United States, Canada, the United Kingdom, Ireland, elsewhere in Europe, Australia, New Zealand, Latin America, Asia, and Israel.^[9]

Research

Abiraterone acetate is under development for the treatment of breast cancer and ovarian cancer and as of March 2018 is in phase II clinical trials for these indications.^[46] It was also under investigation for the treatment of congenital adrenal hyperplasia, but no further development has been reported for this potential use.^[46] An oral ultramicrosize tablet formulation of abiraterone acetate (also known as abiraterone acetate fine particle (AAFP) or submicron abiraterone acetate) with improved bioavailability is in pre-registration in the United States for the treatment of prostate cancer as of April 2018 and has the tentative brand name Yonza.^[47]

PAPER

https://pubs.acs.org/doi/abs/10.1021/op500044p

Improved Procedure for Preparation of Abiraterone Acetate

Mukesh Kumar Madhra*, Hari Mohan Sriram, Murad Inamdar, Mukesh Kumar Sharma, Mohan Prasad, and Sony Joseph

Chemical Research Division, Ranbaxy Research Laboratory, Gurgaon, Haryana 122001, India

Org. Process Res. Dev., 2014, 18 (4), pp 555–558

DOI: 10.1021/op500044p

*E-mail: Mukesh.madhra@ranbaxy.com. Tel: (91-124)4011832.

An improved procedure for the preparation of abiraterone acetate is described. The present process highlights reduced reaction time, isolation with acid–base treatment without involving column chromatography, multiple crystallization and is amenable to large-scale synthesis.

Abiraterone Acetate (1)

1 in 81% yield (1.8 kg). HPLC Purity: 99.72%, Assay: 98.8% (HPLC, w/w). MS: m/z = 392.7 [M + H]⁺. IR (KBr) (cm^–1): 3047, 2936, 1735, 1244, 1035, 801, 714. ¹H NMR (400 MHz, DMSO-d₆): δ 8.58 (s, 1 H), 8.43–8.42 (d, 1 H), 7.76–7.74 (d, 1 H), 7.34–7.31 (dd, 1 H), 6.11 (s, 1 H), 5.38 (s, 1 H), 4.44 (m, 1H), 2.19–2.50 (m, 3H), 1.98–2.08 (m, 6H), 1.39–1.85 (m, 9H), 1.03–1.11 (m, 8H). ¹³C NMR (CDCl₃): δ 170.4, 151.6, 147.9, 147.8, 140.0, 133.6, 132.9, 129.1, 122.9, 122.2, 73.8, 57.4, 50.2, 47.3, 38.1, 36.9, 36.7, 35.1, 31.7, 31.4, 30.3, 27.7, 21.4, 20.8, 19.2, 16.5.

https://pubs.acs.org/doi/suppl/10.1021/op500044p/suppl_file/op500044p_si_001.pdf

Abiraterone (2)

2 (2.88 kg, 72%) as a white solid. HPLC Purity: 99.87%. MS: m/z = 350.3 [M + H]⁺. IR (KBr) (cm^–1): 3236, 3062, 3031, 2931,1596, 1065, 803. ¹H NMR (400 MHz, CDCI₃): δ 8.61 (s, 1 H), 8.44–8.46 (d, 1 H), 7.63–7.65 (d, 1 H), 7.20–7.23 (dd, 1 H), 5.993–5.996 (d, 1 H), 5.38–5.99 (d, 1 H), 3.48–3.54 (m, 1 H), 2.24–2.32 (m, 3H), 1.97–2.10 (m, 3H), 1.47–1.86 (m, 10H), 1.04–1.10 (s, 8H). ¹³C NMR (CDCl₃): δ 16.58, 19.34, 20.88, 30.45, 31.52, 31.63, 31.81, 35.27, 36.71, 37.20, 42.32, 47.34, 50.37, 57.56, 71.62, 121.28, 123.03, 129.24, 132.99, 133.70, 141.21, 147.79, 147.88, 151.68

see supp info

PATENT

CN 201210356832

PATENT

https://patents.google.com/patent/CN103665085A/en

The abiraterone acetate was the ester of formula (Abiraterone acetate) structure.

[0004]

[0005] So far, the search route may abiraterone acetate ester (Abiraterone acetate) are two.

[0006] Patent W09509178, CN 102030798, WO 2006021777, 2006021777, WO2006021776, J.Med.Chem.38,2463-2471,1995, synthetic route reported in the literature like the following formula WO.

[0007]

[0008] The route is DHEA as raw material, with an acetyl group protecting the hydroxyl group, the product obtained is then reacted with trifluoromethanesulfonic anhydride to give triflate product was finally reagent under palladium catalysis, Suzuki coupling reaction with 3-pyridyl diethyl borane, to give an ester of abiraterone acetate.

[0009] Patent GB 2282377,0PPI, 29 (I), 123-134,1997 the reported another method of synthesis.

The synthetic procedure the following formula.

[0010]

[0011] The route is DHEA as raw material, the reaction with hydrazine hydrate, and then reacted with iodine to give the 17-iodo – androsta-5,16-diene–3beta- alcohol, and catalytic agent in the button with Li-yl-pyrazol-diethyl _3_ boron burning Suzuki coupling reaction to give abiraterone, and finally acetylated abiraterone acetate to give abiraterone acetate.

[0012] By comparing the two lines, a synthetic routes can be found with a reagent such as trifluoromethanesulfonic anhydride, 2,6-di-t-butyl-4-methylpyridine and the like are expensive, relatively high chemical costs. In comparison, two synthetic route mild reaction conditions, the reagents are cheap, and therefore have more industrialized prospects. However, according to the synthesis process reported in the literature, the route to industrial production, there are still some technical problems.

[0013] More specifically, to 17- iodo – androsta-5,16-diene–3beta_ when alcohol (2) Synthesis of abiraterone (3) as a raw material for the Suzuki coupling reaction, the solvent is tetrahydrofuran, the solvent high cost; shall reaction refluxed for 4 days, the reaction time is too long. More importantly, when the Suzuki coupling reaction, starting material 17- iodo – male left diene-5,16-ol _3beta_

(2) will react with the impurities abiraterone (3) 4, 4 impurities not removed by recrystallization, can only be purified by column chromatography. If the compound is not 4 Ex, abiraterone prepared by acetylation reaction of abiraterone acetate ester, the impurities will be converted to 4 5 impurities, the impurities by recrystallization 5 likewise not removed, only purified by column chromatography.

[0014]

[0015] The abiraterone acetate ester synthesis, synthesis is reported abiraterone 24h the reaction with acetic anhydride and pyridine at room temperature, the reaction time is too long. The mixture was then evaporated under reduced pressure to be excess acetic anhydride and pyridine, and then crystallized from diethyl ether again, to give the final acetate abiraterone acetate was purified by column chromatography.

[0016] In summary, two synthetic routes reported in the literature of the last two long reaction time and complicated operation, product purification difficult. All this has seriously hampered the industrialization prospects abiraterone acetate esters.It is essential to two synthetic routes to optimize the improvement, in order to achieve the industrial production of abiraterone acetate ester.

] Example 1

Preparation of [0031] 17- (3-pyrazol Li-yl) androsta-5,16-diene-_3beta_ alcohol (abiraterone) of

[0032] A 750ml NMP was added to a 3L three-necked flask, were added with stirring 50gl7_ iodo – androsta-5,16-diene–3beta- alcohol, 88 mg of bis (triphenylphosphine) palladium chloride and diethyl 19.74g yl – (3-pyridyl) borane, and finally adding 345ml 2mol / L Na2CO3 solution. Heating, holding temperature of about 70-80 ° C, TLC monitored the reaction was complete. The reaction was cooled to room temperature, the reaction solution was added 1500ml of water, stirred, filtered and washed with water. Blast drying, 26.3g abiraterone.

[0033] Example 2

[0034] Preparation and purification of abiraterone acetate ester

[0035] The abiraterone 26g 156ml dissolved in pyridine, 52 ml of acetic anhydride was added at room temperature, heating, holding temperature of about 70-80 ° C, the reaction for about 4 h, TLC monitoring of the reaction was complete. The reaction was cooled to room temperature, the ice bath, 560ml of ice water was added to the reaction mixture, the precipitated white solid was stirred 20min, filtered, washed with water. 55 ° C blast drying. The crude product was added to 26ml of ethanol was dissolved by heating to clarify. Water was added 26ml, stirred for lh. Cooled to room temperature and filtered. Blast drying. Abiraterone acetate to give the final acetate 22.lg, HPLC> 99.5%.

[0036] Example 3

Preparation of [0037] 17- (3-pyridyl) androsta-5,16-diene-_3beta_ alcohol (abiraterone) of

[0038] The IlOL NMP was added to a 3L three-necked flask, were added with stirring 7.5kgl7_ iodo – androsta-5,16-diene–3beta- alcohol, 132 g of bis (triphenylphosphine) palladium chloride and 29.6kg two ethyl – (3-pyridyl) borane and finally 500L2mol / L Na2CO3 solution. To maintain the internal temperature of about 70_80 ° C, TLC monitored the reaction was complete. The reaction was cooled to room temperature, 220L of water was added to the reaction mixture, stirred for 30min, filtered, washed with water. Blast drying, 39.2kg abiraterone.

[0039] Example 4

[0040] Preparation and purification of abiraterone acetate ester

[0041] The abiraterone 39kg dissolved in pyridine 230L, 78L of acetic anhydride was added at room temperature, heating, holding temperature of about 70-80 ° C, the reaction for about 4 h, TLC monitoring of the reaction was complete. The reaction was cooled to room temperature, the ice bath, ice water was added to the 840L reaction solution, stirred 30min, filtered, washed with water, 50_55 ° C blast drying. The crude product was added to 39L of ethanol was dissolved by heating to clarify. Water was added 39L, stirred for lh then cooled to room temperature and filtered. Blast drying. Abiraterone acetate to obtain the final ester 33.2kg, HPLC> 99.5% ο

PATENT

https://patents.google.com/patent/WO2013030410A2/en

Abiraterone acetate [17-(3-pyridyl)-5,16-androstadien-33-acetate] is a steroid compound which inhibits selectively and efficiently the enzyme 17-ohydroxylase-C17- 20-lyase, which catalyzes the conversion of dehydroepiandrosterone and androstenedione to testosterone. The inhibition of said enzyme causes a strong decrease of testosterone levels in the patient and therefore this drug is used in the treatment of certain hormone-dependent tumors resistant to chemotherapy such as prostate cancer. This compound has the followin chemical formula:

This product was disclosed for the first time in WO 93/20097, which also provides a synthetic process for its preparation including as last step the reaction of an enol triflate with a pyridine borate by Suzuki coupling (see scheme below). However, this process is not viable in practice, mainly because of the difficulty in preparing the enol trifluorosulfonate at the 17-position 2: this step, apart from proceeding with a poor conversion and low yield, gives place to the impurity tri-unsaturated 3 in a 10% yield, which only may be removed by column chromatography. Further, the product obtained after the subsequent Suzuki coupling must be also purified by column chromatography according to the examples provided therein.

Abiraterone-acetate

The above-mentioned impurity was prevented in later processes (EP 1 781 683 y EP 1 789 432) thanks to the use of alternative bases to that previously employed (i.e. 2,5-ditert-butyl-4-methylpyridine) such as DABCO, DBU or tryethylamine. However, in the sole example described in said documents, whilst the final product is achieved without using any column chromatography, it is obtained in a global yield of scarcely 21 % and shows a purity of only 96.4%.

EP 0 721 461 proposes the use of a vinyl iodide or bromide intermediate instead of the enol triflate, as depicted in the following scheme:

However, the iodo-enol is much less reactive than the triflate in the coupling with the pyridine borane, resulting in long reaction times (48 hours – 4 days) with a part of the starting material unreacted and wherein until a 5% of a dimeric impurity is obtained, which can only be removed by purification by means of reverse phase column chromatograp

Therefore, there is still a need of developing new processes for obtaining 17-(3- pyridyl)-5,16-androstadien-33-ol and related compounds, some of which are of therapeutic interest (e.g. abiraterone acetate) which overcome all or part of the drawbacks associated to the known processes belonging to the state of the art.

PATENT

https://patents.google.com/patent/CN107056871A/en

The chemical name of abiraterone acetate ⑽) -17- (3- pyridyl) – androsta-5,16-diene-3-acetate, by the oral US Centocor Ortho developed CYP17 inhibitor . As anti-cancer drugs on the market April 28, 2011 by the US Food and Drug Administration (FDA) approval, in combination with prednisone therapy with castration-resistant prostate cancer. Trade name Zyitga. Abiraterone acetate is an oral androgen synthesis inhibitors, capable of inhibiting 17a- hydroxylase / C17,20-lyase (CYP17). Clinical results show that abiraterone acetate can significantly prolong patients with advanced prostate cancer include the use of one or both docetaxel-containing chemotherapy but her condition is still deteriorating lives of patients, the risk of death by 35%, and the side effects of drugs is very small, good safety.

[0003] Currently, the synthesis of abiraterone acetate routes are mainly three: (1) dehydroepiandrosterone acetic acid as a starting material, first-butyl-4-methylpyridine 2,6_ di ( esterified by trifluoromethanesulfonic anhydride, then with diethyl under DTBMP) under catalytic bis-triphenylphosphine palladium chloride – acetate proceeded abiraterone acetate Suzuki coupling (2-pyridyl) borane the total yield of the reaction is 48.7%; short reaction step of the process, but after the first-stage reaction a lot of by-products, to be purified by column chromatography, and the double bonds can not be divided by-products, and therefore remains a need for second-stage reaction column chromatography Analysis and purified by recrystallization complicated operation; (2) DHEA as a starting material, a condensation reaction of a hydrazone with hydrazine hydrate, and the presence (TMG) in 1,1,3,3-tetramethylguanidine ene reaction with iodine to generate iodine compound 17- iodo – male left -5,16_ diene -30- alcohol, then in the catalytic bis-triphenylphosphine palladium dichloride and diethyl – (3-pyridyl ) borane was prepared by Suzuki coupling abiraterone abiraterone acetate to give finally acetylated hydroxyl prepared. The total yield of the reaction is 41.5%. This synthesis step is longer, lower yields, and since the active iodides easy to generate high polymer impurities that can not be removed by recrystallization or column chromatography, can only be purified by preparative chromatography to give A in the Suzuki coupling process Long bit acetate pure, can not meet the requirements of industrial production; (3) in the method (1) was treated with triethylamine instead of DTBMP, reduces the formation of byproducts double bond, then after the reaction the remaining starting material was recrystallized removed. This reaction increases the process steps and the purity of the final product was only 96.4%, the drug does not meet the quality standards.

Example 1

[0021] One method of synthesis of abiraterone acetate, comprising the steps of:

[0022] A, in a 100ml round bottom flask were sequentially added 0 • 95g (5mmol) of p-toluenesulfonyl chloride, 15 mL of toluene, sufficiently stirred to dissolve, to obtain X-solution, (solution cooled to 15 ° C X) at 15 ° C under a slow was added dropwise by molar ratio of 1: 2 was added 1.5mL (lOmmol) 80% hydrazine hydrate to the solution X, dropwise within 5min; 3〇min reaction was continued, white precipitate appears in the flask. TLC analysis of the reaction end point is determined. After completion of the reaction, cold water was added 3〇mllO ° C., Stirred, filtered off with suction, the filter cake was then washed with purified water 3-5 times, and dried to obtain a white crystalline p-toluene sulfonyl hydrazide billion • 82g, 86.3% yield ( literature values: yield 92%).

[0023] B, DHEA -17- Synthesis of p-toluenesulfonyl hydrazone

[0024] In a 100ml round bottom flask were sequentially added in square • 75g CMmo 1) dehydroepiandrosterone (DHEA), 25 mL of methanol, 0.81 g sufficiently stirred to dissolve the p-toluene sulfonyl hydrazide, rt (15_25 ° C), was added O.lmL 0.2 mol .L-1 sulfuric acid, 60 ° C in an oil bath at reflux for 2h, TLC analysis of the reaction end point is determined. After completion of the reaction, the solvent was largely removed by rotary evaporation, a heavy white precipitate appeared in the flask. Was added 30mL of water, filtered off with suction, the filter cake was then washed with purified water 3-5 times to remove water at one thousand bake 50 ° C, to give a white solid 1.27g i.e. -17- Dehydroepiandrosterone p-toluenesulfonyl hydrazone, yield 81.4%.

[0025] C, 17- (3- pyridyl) androsta-5,16-diene–30- _ Synthesis of alcohol

[0026] In a 100ml round bottom flask was added 0.91g (2mmol) -17- Dehydroepiandrosterone p-toluenesulfonyl hydrazone, 25mLl, 4- dioxane, 〇.27g (3 mmol of) lithium tert-butoxide, 0_012g (0.013mmol) Pd2 (dba) 3,0.023g stirred for five minutes to fully dissolve the (0_005mmol) Xphos, at room temperature, wherein Pd2 (dba) 3 were added under nitrogen, and then quickly added 0.39g (2.5mmol) 3- bromo pyridine, 95 ° C oil bath reactor 12h, TLC analysis of the reaction end point is determined. After the reaction, ice water was added 30mL0 ° C, thoroughly shaken, ethyl acetate was added 20mL of acetic acid, liquid separation, was extracted with ethyl acetate (1 〇ml each, extracted three times) and the combined organic phase was dried over anhydrous sodium sulfate (by lg / ml was added over anhydrous sodium ratio) sulfate, filtered, and the filtrate rotary evaporated to give a pale yellow solid which was recrystallized from n-hexane (20ml) to give a white solid that is 17- (3-pyridyl) – male stay -5, 16- -3P- diene alcohols, a yield of 42.6%.

[0027] D, Synthesis of abiraterone acetate

[0028] In 0.39gl7- successively added 100mL round bottom flask (3-pyridyl) – androsta-5,16-diene-fir -3 – ol, 3〇111 dagger diethyl ether, 0.15mL (0.25mmol) triethylamine amine, are hook stirring, was slowly added dropwise 0.3 mL (2mm〇l) of acetyl chloride, the reaction stirred at room temperature Jiao 3h, TLC analysis of the reaction end point is determined. The mixture was then suction filtered, the filtrate was decolorized with charcoal, rotary evaporation, to give a white solid, i.e. abiraterone acetate product yield of 80.6%.

[0029] Example 2

[0030] A, in a 100ml round bottom flask were added 1. (^ (5.311111101) p-toluenesulfonyl chloride, 15 mL of toluene, sufficiently stirred to dissolve to give the solution X, at 15 ° C was slowly added dropwise 1 • 7mL (12mmol) X 80% hydrazine hydrate to the solution in dropwise within 5min; reaction was continued for 30min, a white precipitate appeared .TLC analysis to determine the end of the reaction after the completion of the reaction flask was added 30ml 10 ° C water with stirring, suction filtered, then the filter cake. washed 3-5 times purified water, was dry, i.e., p-toluenesulfonyl hydrazide to give white crystals 0.93 g, 86.7% yield (literature: yield 92%).

[0031] B, DHEA -17- Synthesis of p-toluenesulfonyl hydrazone

[0032] successively added 0 • 97g (3mmo 1) dehydroepiandrosterone (DHEA) in a 100ml round bottom flask, 25 mL of methanol, 1 • 08g p-toluene sulfonyl hydrazide, fully dissolved with stirring at room temperature, was added O.lmL 0.2mol • L_1 sulfuric acid, 60 ° C in an oil bath at reflux for 2h, TLC analysis of the reaction end point is determined. After completion of the reaction, the solvent was largely removed by rotary evaporation, a heavy white precipitate appeared in the flask. Was added 30mL of water, filtered off with suction, the filter cake was then washed with purified water 3-5 times, 5 (TC drying under removal of water, to give a white solid 1.43g i.e. Dehydroepiandrosterone p-toluenesulfonyl hydrazone -17- yield 80.9%.

[0033] C, 17- (3- pyridyl) -30- _ male left diene -5,16-ol Synthesis

[0034] In a 100ml round bottom flask was added in 1.32g (3 mmol of) -17- Dehydroepiandrosterone p-toluenesulfonyl hydrazone, 25mLl, 4- dioxane, 0.35g (4mmol) of lithium t-butoxide, 0.012g (0.013_ol) Pd2 (dba) 3,0.023g stirring for five minutes under fully dissolved (0.005mmol) Xphos, at room temperature, wherein Pd2 (dba) 3 were added under nitrogen, and then quickly added 0.48g (3mmol) 3- bromo pyridine, 80 ° C oil bath and the reaction 19h, TLC analysis of the reaction end point is determined. After the reaction, 30mL of ice water was added, shaken well, 2〇mL ethyl acetate was added, liquid separation, was extracted with 30ml ethyl acetate (10ml each, extracted three times) and the combined organic phases were scaled lg / ml was added anhydrous dried over sodium sulfate, filtered, and the filtrate was rotary evaporated to give a pale yellow solid which was recrystallized from 20ml of n-hexane to give a white solid that is 17- (3-pyridyl) – androsta-5,16-diene–3P- alcohol, 42 • 6% yield.

[0035] D, Synthesis of abiraterone acetate

[0036] In 0.41gl7- successively added 100mL round bottom flask (3-pyridyl) -! -33- androst-5,16-diene-ol, ^ 3〇 diethyl ether, 0.2mL (0 • 3 round 〇1 ) of triethylamine, stir, slowly added dropwise 0.3mL (2mmol) of acetyl chloride, the reaction was stirred at room temperature for 3h, TLC analysis of the reaction end point is determined. The mixture was then suction filtered, the filtrate was decolorized with charcoal, rotary evaporation, to give a white solid, i.e. abiraterone acetate product yield of 81 • 2%.

[0037] Example 3

[0038] A, were added in a 100ml round-bottomed flask 1.08g (5.7mmo 1) p-toluenesulfonyl chloride, 15 mL of toluene, sufficiently stirred to dissolve to give the solution X, at 15 ° C was slowly added dropwise 1 • 8mL (13mmo 1 ) of 80% hydrazine hydrate to the solution X, dropwise within 5min; reaction was continued for 30min, a white precipitate appeared in the flask. TLC analysis of the reaction end point is determined. After completion of the reaction, water 30ml 10 ° C with stirring, filtered off with suction, the filter cake was then washed with purified water 3-5 times, and dried to obtain a white crystalline p-toluene sulfonyl hydrazide 0.94g, 85.6% (Yield literature values: yield 92%).

[0039] B, DHEA -17- Synthesis of p-toluenesulfonyl hydrazone

[0040] 1 • 18g were added in a 100ml round bottom flask (3.3 mmol) Dehydroepiandrosterone, 25 mL of methanol, 1.28 g of p-toluene sulfonyl hydrazide, fully dissolved with stirring at room temperature, was added O.lmL 0.2mol • I / a sulfate, an oil bath at reflux for 2h, TLC analysis of the reaction end point is determined. After completion of the reaction, the solvent was largely removed by rotary evaporation, a heavy white precipitate appeared in the flask. Was added 30mL of water, filtered off with suction, the filter cake was then washed with purified water 3-5 times, 5 (TC drying under removal of water, to give a white solid 1.51g i.e. Dehydroepiandrosterone p-toluenesulfonyl hydrazone -17- yield 80.9%.

[0041] C, 17- (3- pyridyl) – androst _5,16- diene synthetic alcohols -3P-

[0042] Add l_45g (3.2mmol) -17- Dehydroepiandrosterone p-toluenesulfonyl hydrazone, 25mLl, 4- dioxane, 0 • 35g in 100ml round bottom flask (4mmol) of lithium tert-butoxide, 0.012 g (0.013mmol) Pd2 (dba) 3,0.023g (0.005ramol) Xphos, fully dissolved after stirred at room temperature for five minutes, wherein Pd2 (dba) 3 were added under nitrogen, then added rapidly 0 • 60g (4mmol) 3 – bromopyridine, 120 ° C oil bath and the reaction 9h, TLC analysis of the reaction end point is determined. After the reaction, 30mL of ice water was added, shaken well, was added 20mL of ethyl acetate, separated, extracted with 30ml ethyl acetate (10ml each, extracted three times) and the combined organic phases were scaled lg / ml was added over anhydrous sodium to intervene sulfate, filtered, and the filtrate rotary evaporated to give a pale yellow solid which was recrystallized from burning 2〇ml n-hexyl, i.e. 17_ to give a white solid (3-Jie ratio piperidyl) – androst -5,16_ diene -30- alcohols, a yield of 43.1%.

[0043] D, Synthesis of abiraterone acetate

[0044] successively added in a round bottom flask i〇〇mL 0.52gl7- (3- pyridyl) -! -30- androst-5,16-diene-ol, ^ 3〇 diethyl ether, 0.25mL (0.36mmol) triethylamine, stir, slowly added dropwise 0.35 mL (2.2 mmol) of acetyl chloride, the reaction was stirred at room temperature for 3h, TLC analysis of the reaction end point is determined. The mixture was then suction filtered, the filtrate was decolorized with charcoal, rotary evaporation, to give a white solid, i.e. abiraterone acetate product yield of 81.8%.

[0045] In each of the above embodiments, the improved synthetic route abiraterone acetate to DHEA as raw materials by the condensation of p-toluenesulfonyl hydrazide, and then reacted with 3-bromopyridine coupling occurs, acetylation, etc. 3 target product was synthesized from abiraterone acetate, 43.4% overall yield.Route and the mild reaction conditions, readily available and inexpensive raw materials, low production cost.

PAPER

Pharmaceutical Chemistry Journal

September 2016, Volume 50, Issue 6, pp 404–406| Cite as

Four-Step Synthesis of Abiraterone Acetate from Dehydroepiandrosterone

https://link.springer.com/article/10.1007/s11094-016-1459-1

Balaev, A.N., Gromyko, A.V. & Fedorov, V.E. Pharm Chem J (2016) 50: 404. https://doi.org/10.1007/s11094-016-1459-1

Syn 1

J Med Chem 1995,38(13),2463

Treatment of dehydroepiandrosterone 3-acetate (I) with triflic anhydride and 2,6-di-tert-butyl-4-methylpyridine provided the desired enol triflate (III) along with some 3,5-diene (II), which were separated by column chromatography. Subsequent coupling of triflate (III) with pyridylborane (IV) using bis(triphenylphosphine)- palladium(II) chloride as the catalyst afforded the (3-pyridyl)androstadiene (V), which after hydrolysis of the acetate ester with NaOH provided the target compound.

Abiraterone

Substances Referenced in Synthesis Path

CAS-RN	Formula	Chemical Name	CAS Index Name
853-23-6	C21H30O3	dehydroepiandrosterone-3-acetate	Androst-5-en-17-one, 3-(acetyloxy)-, (3β)-
89878-14-8	C9H14BN	diethyl (3-pyridyl)borane	Pyridine, 3-(diethylboryl)-
C26H33NO2	(3β)-acetoxy-17-(3-pyridyl)androsta-5,16-diene

Trade Names

Country	Trade Name	Vendor	Annotation
EU	Zytiga	Janssen Cilag, 2011
USA	Zytiga	Johnson & Johnson, 2011

Formulations

• tabs. 250 mg

References

• • Potter, G. A. et al., J. Med. Chem., (1995) 38, 2463.
  • US 5 604 213 (British Technology Group; 18.2.1997; appl. 30.9.1994; GB-prior. 31.3.1992).
  • EP 633 893 (Brit. Technology Group; 18.1.1995; appl. 15.3.1993; GB-prior. 31.3.1992).
  • WO 9 320 097 (Brit. Technology Group; 14.10.1993; appl. 15.3.1993; GB-prior. 31.3.1992).

• large scale synthesis of acetate:

  • Potter, G. A. et al., Org. Prep. Proced. Int., (1997) 29, 123.

CLIP

Abiraterone acetate, the active ingredient of ZYTIGA is the acetyl ester of abiraterone. Abiraterone is an inhibitor of CYP17 (17α-hydroxylase/C17,20-lyase). Each ZYTIGA tablet contains either 250 mg or 500 mg of abiraterone acetate. Abiraterone acetate is designated chemically as (3β)-17-(3-pyridinyl) androsta-5,16-dien-3-yl acetate and its structure is:

Abiraterone acetate is a white to off-white, non-hygroscopic, crystalline powder. Its molecular formula is C₂₆H₃₃NO₂ and it has a molecular weight of 391.55. Abiraterone acetate is a lipophilic compound with an octanol-water partition coefficient of 5.12 (Log P) and is practically insoluble in water. The pKa of the aromatic nitrogen is 5.19.

ZYTIGA tablets are available in 500 mg film-coated tablets, 250 mg film-coated tablets and 250 mg uncoated tablets with the following inactive ingredients:

• 500 mg film-coated tablets: colloidal silicon dioxide, croscarmellose sodium, hypromellose, lactose monohydrate, magnesium stearate, silicified microcrystalline cellulose, and sodium lauryl sulfate. The coating, Opadry® II Purple, contains iron oxide black, iron oxide red, polyethylene glycol, polyvinyl alcohol, talc, and titanium dioxide.
• 250 mg film-coated tablets: colloidal silicon dioxide, croscarmellose sodium, lactose monohydrate, magnesium stearate, microcrystalline cellulose, povidone, and sodium lauryl sulfate. The coating, Opadry® II Beige, contains iron oxide red, iron oxide yellow, polyethylene glycol, polyvinyl alcohol, talc, and titanium dioxide.
• 250 mg uncoated tablets: colloidal silicon dioxide, croscarmellose sodium, lactose monohydrate, magnesium stearate, microcrystalline cellulose, povidone, and sodium lauryl sulfate.

PAPER

A CONVENIENT, LARGE-SCALE SYNTHESIS OF ABIRATERONE ACETATE [3β-ACETOXY-17-(3-PYRIDYL)ANDROSTA-5,16-DIENE], A POTENTIAL NEW DRUG FOR THE TREATMENT OF PROSTATE CANCER

Organic Preparations and Procedures International , The New Journal for Organic Synthesis , Volume 29, 1997 – Issue 1

https://www.tandfonline.com/doi/abs/10.1080/00304949709355175

• 10.1080/00304949709355175
• 3P-Acetoxy- 17-(3-pyridyl)androsta-5,16-diene (abiraterone acetate, 5) is a pro-drug for 17- (3-pyridyl)androsta-5,16-dien-3P-ol (abiraterone, 4), a potent inhibitor of human cytochrome P450,,, (steroidal 17a-hydroxylase-C,,,,,-lyase).’ This enzyme is a potential target for a drug designed to treat hormone-dependent prostatic carcinoma, and 5 has been approved for clinical trial in patients with this cancer. In connection with this trial, a method amenable to the synthesis of 5 on a large scale (ca. 100 g or more) was required. The present paper reports such a synthesis.

The key step in the previously reported’ synthesis of 5 was the palladium-catalysed crosscoupling reaction between diethyl(3-pyridy1)borane and the 17-en01 triflate derived from the 3-acetate of dehydroepiandrosterone 1. The procedure has potential drawbacks as a method for large-scale synthesis. Aside from the use of the expensive and noxious triflic anhydride, the formation of the enol triflate requires the expensive hindered base 2,6-di-tert-b~tyl-4-methylpyridine.~ Further, it was accompanied by some elimination of acetic acid to give androsta-3,5,16-trien- 17-yl triflate which required chromatographic separation from the desired product, and contributed to reducing its isolated yield from the acetate of 1 to a moderate 58%. These problems prompted consideration of an altemative steroidal precursor suitable for the cross-coupling reaction. It occurred to us that the vinyl iodide 3 might provide a viable alternative to an enol triflate in the palladium catalyzed cross-coupling step. Such steroidal vinyl iodides are easily and cheaply obtained via the corresponding 17-h~drazones.’-~ The synthesis of 3 iself from the hydrazone3 2 by oxidation with iodine in the presence of a hindered guanidine base has been optimi~ed~.~ to obtain the product on a small scale (0.13 g) in 95% yield. We were able to repeat this reaction on a large scale and obtain a similar yield of 3. The palladium catalysed cross-coupling reaction of 3 with diethyl(3-pyridy1)borane proceeded without the need to protect the 3-hydroxyl function to give 4, whereas the use of an enol triflate in the coupling reaction does not conveniently allow this option. However, coupling with the iodide was much slower, requiring 4 days at 80″ as compared with the 1 hr required when an enol triflate precursor was used.’ RO A ‘ OR Recrystallization of the crude 4 obtained by the foregoing procedure gave a product with melting point lower than that previously reported,’ and TLC revealed a less mobile contaminant that was not removed by further recrystallization. The crude product was therefore acetylated to give the crude target compound 5 contaminated with a by-product. This by-product was 6, formed from a precursor 7 present as a contaminant in crude 4. The prolonged reaction time required for the cross-coupling reaction using the vinyl iodide 3 had enabled a Heck-type reaction7 to occur between the initial product 4 and the bis(tripheny1phosphine)- palladium derivative of 3 to form 7. The very recently reported8 palladium-catalysed dimerisation of 17-i0dod’~-steroids to give 16,17′-coupled products provides a precedent for this side-reaction. Whereas column chromatography on silica-gel of crude 5 afforded pure 6, which was eluted first, compound 6 contaminated later fractions and could not be completely removed from 5 by recrystallization. However subsequent reverse phase chromatography allowed the complete separation of the now faster eluting 5 from 6, and recovery of >lo0 g of pure 5 by batchwise chromatography of the crude product. The by-product 6 was deacetylated to give 7, the contaminant present in 4 prepared by the present route. Neither of the new compounds 6 and 7 was appreciably inhibitory towards the human cytochrome P450,,, (S. E. Barrie, personal communication). The availability of pure 7 affords the option of exploring the purification of 4 prior to acetylation. However, for chromatographic purification, the greater solubilities of 5 and 6 in suitable organic solvents compared with their non-acetylated counterparts favor the present choice of purification after acetylation.

3P-Acetoxy-17-(3-pyridyl)androsta-5,16-diene ( 5) and 3~-acetoxy-16-(3~-acetoxyandrosta-5,16- dien-17-yl)-17-(3-pyridyl)androsta-5,16-diene (6).- To a stirred suspension of the product from the foregoing reaction (36.5 g, 0.104 mol) in dry pyridine (200 mL) in a 500 mL round-bottomed flask was added acetic anhydride (75 mL) and the mixture stirred at room temperature for 24 hrs. The pyridine and excess acetic anhydride was removed on a rotary evaporator, initially at water pump pressure with the water bath at 70 “, and finally under high vacuum at 80″ for 30 min. The resulting oil was dissolved in Et,O (500 mL), washed with saturated aqueous NaHCO, (2 x 200 mL), dried (N%CO,), and concentrated to an oil which crystallised on standing. The crude 5 was partly purified by preparative flash chromatography on silica gel using a 9 cm diameter column, eluting with dichloromethane. A by-product (6) eluted first and was followed by fractions variously enriched in 5. The foregoing reaction and purification procedure was carried out a total of four times, thus using a total of 146 g (0.41 8 mol) of crude 4. The dichloromethane fractions containing the least by-product were combined and concentrated. Recrystallisation from hexane afforded product (1 08 g) consisting of 5 containing 6.8% w/wof 6 as impurity as determined by analytical HPLC. The more contaminated fractions similarly afforded product (25 g) containing 21 3% w/w of 6 (we thank Dr C. P. Quarterman, Aston Molecules Ltd, Birmingham U.K. for these analyses). A pure sample of 6 (4 g) was isolated from the combined initial fractions as pale yellow crystals, mp. 269-270” (from hexane); IR 1732 cm-‘ (GO str); ‘H-NMR: 6 0.85 (s, 3, H-18′), 1.02, 1.04 (2s, 6, H-19,19′), 1.06 (s, 3, H-18), 2.034, 2.039 (2s, 6, CH,CO), 4.59 (2m, 2, H-3,3’), 5.13 (s, 1, H-16), 5.39 (dd, 2, H-6,6), 7.62 (dd, 1, Js,4 = 8.0 Hz, Js,6 = Anal. Calcd for C,,H,,NO,: C, 80.18; H, 8.73; N, 1.99. Found: C, 80.19; H, 8.78; N, 1.95

The crude 5 was purified by reverse-phase column chromatography. A solution of material from the 108 g batch (10 g) in a hot mixture of acetonitrile (200 mL) and methanol (40 mL) was allowed to cool and filtered. The filtrate was applied to a 10 cm diameter column (500 g) of LiChroprep@ RP-8 reverse-phase C, packing Art. No. 9324. The column was eluted with acetonitrile-0.05 M ammonium acetate (20: 1) with a flow rate of 25 amin and 500 mL fractions were collected and analysed by analytical HPLC (see below). Fractions 4-10 contained pure 5. After a further two fractions, the eluant was changed to acetonitrile-acetic acid (20:l) and pure 6 was completely eluted in fractions 16-19. In 3 subsequent runs using the same column, 25 g portions of the same batch of crude 5 were each dissolved in a mixture of hot acetonitrile (350 mL) and methanol (100 mL) and processed as before. Fractions 2-7 contained pure 5 and, following the change of solvent, fractions 8-12 contained 6. The four eluates containing 5 were combined and recrystallised from acetonitrile (1200 mL) to give pure 5 (57.5 g), mp. 146-148″, lit.’ mp. 144-145′, in which 6 was not detected by analytical HPLC (for procedure, see below) at the limit of detection (<0,05% w/w 6). Anal. Calcd for C,,H,,NO,: C, 79.75; H, 8.50; N, 3.58. Found: C, 79.73; H, 8.48; N, 3.62

Further material (14 g) from the 108 g batch was combined with a portion (22 g) of the more impure 25 g batch and the total of 36 g was chromatographed in one batch as above, again giving complete separation of 5 from 6. Recrystallisation from acetonitrile (600 mL) gave a further 28.5 g of 5 of purity equal to the foregoing crop of 57.5 g 5. Concentration of the combined mother liquors from these crops followed by addition of water (MeCN:&O, 12:l v/v) gave further pure 5 (17.5 g). The total recovery of pure 5 was therefore 103.5 g (36% based on 3). The spectroscopic data (NMR, IR, and MS) of the final products from this procedure were identical with those reported for the product obtained by the route previously described.’

Procedure for Analysis of Purity of Batches of 5 Using Analytical HPLC.- The eluant was acetonitrile-0.05M ammonium acetate and the flow rate 1.5 mumin. Components were monitored either by fluorescence detection (excitation wavelength hex 262 nm, emission wavelength hem 353 nm) or by UV detection (254 nm). Typical retention times were: for 5,225 sec; for 6, 1162 sec. For analysis of crystalline products, a solution (5 mg/mL) in acetonitrile was diluted 50 fold to 100 pg/mL and 100 pl of this solution was injected onto the column.

1 G. A. Potter, S. E. Barrie, M. Jarman and M. G. Rowlands, J. Med. Chem., 38,2463 (1995).

References

1. ^ Jump up to:^{a b c d e f g h i j k l m n o p q} Janssen Biotech, Inc. “ZYTIGA Prescribing Information”(PDF). Horsham, PA: U.S. Food and Drug Administration.
2. ^ Jump up to:^a b c d “Meeting Library – Meeting Library”. meetinglibrary.asco.org.
3. ^ Jump up to:^a b c d Benoist GE, Hendriks RJ, Mulders PF, Gerritsen WR, Somford DM, Schalken JA, van Oort IM, Burger DM, van Erp NP (November 2016). “Pharmacokinetic Aspects of the Two Novel Oral Drugs Used for Metastatic Castration-Resistant Prostate Cancer: Abiraterone Acetate and Enzalutamide”. Clin Pharmacokinet. 55 (11): 1369–1380. doi:10.1007/s40262-016-0403-6. PMC 5069300 . PMID 27106175.
4. Jump up^ Potter et al. J. Med. Chem., 1995, 38 (13), pp 2463–2471
5. ^ Jump up to:^a b Scowcroft H (2011-09-21). “Where did abiraterone come from?”. Cancer Research UK. Retrieved 2011-09-28.
6. ^ Jump up to:^a b “Drugs@FDA – FDA Approved Drug Products – Zytiga”. http://www.fda.gov. United States Food and Drug Administration. Retrieved 4 March 2016.
7. ^ Jump up to:^a b “FDA approves Zytiga for late-stage prostate cancer” (Press release). United States Food and Drug Administration. 2011-04-28.
8. Jump up^ NCI Staff. “Abiraterone Approved for Earlier Use in Men with Metastatic Prostate Cancer”. U.S. Department of Health and Human Services, National Institutes of Health, National Cancer Institute.
9. ^ Jump up to:^{a b c d e} “Abiraterone”. Drugs.com.
10. Jump up^ “Generic Zytiga Availability”. Drugs.com.
11. ^ Jump up to:^a b c “Zytiga (abiraterone acetate) tablet [Janssen Biotech, Inc.]”. DailyMed. Janssen Biotech, Inc. September 2013. Retrieved 24 January 2014.
12. ^ Jump up to:^a b c d “Zytiga: EPAR – Product Information” (PDF). European Medicines Agency. Janssen-Cilag International N.V. 29 October 2013. Retrieved 24 January 2014.
13. ^ Jump up to:^a b c “Zytiga 250 mg tablets – Summary of Product Characteristics”. electronic Medicines Compendium. Janssen-Cilag Ltd. 21 January 2014. Retrieved 24 January 2014.
14. ^ Jump up to:^a b c “Zytiga abiraterone acetate product information” (PDF). TGA eBusiness Services. Janssen-Cilag Pty Ltd. 1 March 2012. Retrieved 24 January 2014.
15. Jump up^ “Pharmaceutical Benefits Scheme – Abiraterone”. Pharmaceutical Benefits Scheme. Retrieved 24 January 2014.
16. Jump up^ Clinical trial number NCT00638690 for “Abiraterone Acetate in Castration-Resistant Prostate Cancer Previously Treated With Docetaxel-Based Chemotherapy” at ClinicalTrials.gov
17. Jump up^ de Bono JS, Logothetis CJ, Molina A, Fizazi K, North S, Chu L, Chi KN, Jones RJ, Goodman OB, Saad F, Staffurth JN, Mainwaring P, Harland S, Flaig TW, Hutson TE, Cheng T, Patterson H, Hainsworth JD, Ryan CJ, Sternberg CN, Ellard SL, Fléchon A, Saleh M, Scholz M, Efstathiou E, Zivi A, Bianchini D, Loriot Y, Chieffo N, Kheoh T, Haqq CM, Scher HI (May 2011). “Abiraterone and increased survival in metastatic prostate cancer”. The New England Journal of Medicine. 364 (21): 1995–2005. doi:10.1056/NEJMoa1014618. PMC 3471149 . PMID 21612468.
18. Jump up^ Clinical trial number NCT00887198 for “Abiraterone Acetate in Asymptomatic or Mildly Symptomatic Patients With Metastatic Castration-Resistant Prostate Cancer” at ClinicalTrials.gov
19. Jump up^ “BTG and Ortho Biotech’s Prostate Cancer Trial Unblinded”. Genetic Engineering & Biotechnology News. 2010-09-10. Retrieved 2011-05-26.
20. Jump up^ Ryan CJ, Smith MR, de Bono JS, Molina A, Logothetis CJ, de Souza P, Fizazi K, Mainwaring P, Piulats JM, Ng S, Carles J, Mulders PF, Basch E, Small EJ, Saad F, Schrijvers D, Van Poppel H, Mukherjee SD, Suttmann H, Gerritsen WR, Flaig TW, George DJ, Yu EY, Efstathiou E, Pantuck A, Winquist E, Higano CS, Taplin ME, Park Y, Kheoh T, Griffin T, Scher HI, Rathkopf DE (January 2013). “Abiraterone in metastatic prostate cancer without previous chemotherapy”. The New England Journal of Medicine. 368 (2): 138–48. doi:10.1056/NEJMoa1209096. PMC 3683570 . PMID 23228172.
21. ^ Jump up to:^{a b c d e} “Zytiga prescribing information” (pdf). Janssen Biotech. May 2012. Retrieved 4 March 2016.
22. ^ Jump up to:^{a b c d e} “Zytiga (abiraterone) dosing, indications, interactions, adverse effects, and more”. Medscape Reference. WebMD. Retrieved 24 January 2014.
23. ^ Jump up to:^a b Neidle S (30 September 2013). Cancer Drug Design and Discovery. Academic Press. pp. 341–342. ISBN 978-0-12-397228-6.
24. Jump up^ Fernández-Cancio, Mónica; Camats, Núria; Flück, Christa E.; Zalewski, Adam; Dick, Bernhard; Frey, Brigitte M.; Monné, Raquel; Torán, Núria; Audí, Laura (2018-04-29). “Mechanism of the Dual Activities of Human CYP17A1 and Binding to Anti-Prostate Cancer Drug Abiraterone Revealed by a Novel V366M Mutation Causing 17,20 Lyase Deficiency”. Pharmaceuticals. 11 (2): 37. doi:10.3390/ph11020037.
25. Jump up^ Attard G, Belldegrun AS, de Bono JS (December 2005). “Selective blockade of androgenic steroid synthesis by novel lyase inhibitors as a therapeutic strategy for treating metastatic prostate cancer”. BJU International. 96 (9): 1241–6. doi:10.1111/j.1464-410X.2005.05821.x. PMID 16287438.
26. ^ Jump up to:^a b c Small EJ (November 2014). “Can targeting the androgen receptor in localized prostate cancer provide insights into why men with metastatic castration-resistant prostate cancer die?”. Journal of Clinical Oncology. 32 (33): 3689–91. doi:10.1200/JCO.2014.57.8534. PMID 25311216. Abiraterone acetate is a prodrug for abiraterone, a CYP17 inhibitor, which has the capacity to lower serum testosterone levels to less than 1 ng/dL (compared with levels closer to 20 ng/dL that are achieved with conventional ADT).19 […] Relative to LHRHa alone, the addition of abiraterone resulted in an 85% decline in dihydrotestosterone (DHT) levels, a 97% to 98% decline in dehydroepiandrosterone (DHEA) levels, and a 77% to 78% decline in androstenedione levels.
27. Jump up^ Tindall DJ, James M (20 April 2009). Androgen Action in Prostate Cancer. Springer Science & Business Media. pp. 748–. ISBN 978-0-387-69179-4.
28. Jump up^ Yin L, Hu Q (January 2014). “CYP17 inhibitors–abiraterone, C17,20-lyase inhibitors and multi-targeting agents”. Nature Reviews. Urology. 11 (1): 32–42. doi:10.1038/nrurol.2013.274. PMID 24276076.
29. Jump up^ Malikova, Jana; Brixius-Anderko, Simone; Udhane, Sameer S.; Parween, Shaheena; Dick, Bernhard; Bernhardt, Rita; Pandey, Amit V. (2017). “CYP17A1 inhibitor abiraterone, an anti-prostate cancer drug, also inhibits the 21-hydroxylase activity of CYP21A2”. The Journal of Steroid Biochemistry and Molecular Biology. 174: 192–200. doi:10.1016/j.jsbmb.2017.09.007. ISSN 1879-1220. PMID 28893623.
30. Jump up^ Udhane, Sameer S.; Dick, Bernhard; Hu, Qingzhong; Hartmann, Rolf W.; Pandey, Amit V. (2016). “Specificity of anti-prostate cancer CYP17A1 inhibitors on androgen biosynthesis”. Biochemical and Biophysical Research Communications. 477 (4): 1005–1010. doi:10.1016/j.bbrc.2016.07.019. ISSN 1090-2104. PMID 27395338.
31. ^ Jump up to:^a b Li Z, Bishop AC, Alyamani M, Garcia JA, Dreicer R, Bunch D, Liu J, Upadhyay SK, Auchus RJ, Sharifi N (July 2015). “Conversion of abiraterone to D4A drives anti-tumour activity in prostate cancer”. Nature. 523 (7560): 347–51. doi:10.1038/nature14406. PMC 4506215 . PMID 26030522.
32. ^ Jump up to:^a b Li Z, Alyamani M, Li J, Rogacki K, Abazeed M, Upadhyay SK, Balk SP, Taplin ME, Auchus RJ, Sharifi N (May 2016). “Redirecting abiraterone metabolism to fine-tune prostate cancer anti-androgen therapy”. Nature. 533 (7604): 547–51. doi:10.1038/nature17954. PMID 27225130.
33. ^ Jump up to:^a b c Capper CP, Larios JM, Sikora MJ, Johnson MD, Rae JM (May 2016). “The CYP17A1 inhibitor abiraterone exhibits estrogen receptor agonist activity in breast cancer”. Breast Cancer Research and Treatment. 157 (1): 23–30. doi:10.1007/s10549-016-3774-3. PMID 27083183.
34. Jump up^ Alesini D, Iacovelli R, Palazzo A, Altavilla A, Risi E, Urbano F, Manai C, Passaro A, Magri V, Cortesi E (2013). “Multimodality treatment of gynecomastia in patients receiving antiandrogen therapy for prostate cancer in the era of abiraterone acetate and new antiandrogen molecules”. Oncology. 84 (2): 92–9. doi:10.1159/000343821. PMID 23128186.
35. Jump up^ Figg WD, Chau CH, Small EJ (14 September 2010). Drug Management of Prostate Cancer. Springer Science & Business Media. p. 97. ISBN 978-1-60327-829-4.
36. Jump up^ Rosenthal L, Burchum J (17 February 2017). Lehne’s Pharmacotherapeutics for Advanced Practice Providers – E-Book. Elsevier Health Sciences. p. 936. ISBN 978-0-323-44779-9.
37. Jump up^ Yip CK, Bansal S, Wong SY, Lau AJ (April 2018). “Identification of Galeterone and Abiraterone as Inhibitors of Dehydroepiandrosterone Sulfonation Catalyzed by Human Hepatic Cytosol, SULT2A1, SULT2B1b, and SULT1E1”. Drug Metabolism and Disposition. 46 (4): 470–482. doi:10.1124/dmd.117.078980. PMID 29436390.
38. Jump up^ “A new way to treat prostate cancer: The story of abiraterone”. The Institute of Cancer Research. 2012-09-10. Retrieved 2012-11-12.
39. Jump up^ “Abiraterone Acetate (CB7630)”. Cougar Biotechnology. Archived from the original on 7 September 2008. Retrieved 2008-08-20.
40. Jump up^ “Johnson & Johnson Announces Definitive Agreement to Acquire Cougar Biotechnology, Inc” (Press release). Cougar Biotechnology. 2009-05-11. Archived from the original on 29 May 2009. Retrieved 2009-06-03.
41. Jump up^ “FDA Approval for Abiraterone Acetate”. U.S. Department of Health and Human Services, National Institutes of Health, National Cancer Institute.
42. Jump up^ “EMA assessment of Zytiga (abiraterone)”. European Medicines Agency.
43. Jump up^ “Prostate cancer (metastatic, castration resistant) – abiraterone (following cytoxic therapy): final appraisal determination guidance” (PDF). NICE guidance. 15 May 2012. Archived from the original (PDF) on February 19, 2013.
44. Jump up^ “NICE technology appraisal guidance [TA259]”. NICE guidance. June 2012.
45. Jump up^ “NICE appraisal of earlier treatment with abiraterone for prostate cancer”. NICE press release. 14 August 2014.
46. ^ Jump up to:^a b c “Abiraterone acetate – Johnson & Johnson”. Adis Insight.
47. Jump up^ “Abiraterone acetate – Churchill Pharmaceuticals”. Adis Insight.

External links

• Zytiga (abiraterone acetate) – Official website
• Abiraterone acetate – AdisInsight

Abiraterone acetate

Clinical data
Trade names	Zytiga, others
Synonyms	CB-7630; JNJ-212082; 17-(3-Pyridinyl)androsta-5,16-dien-3β-ol acetate
AHFS/Drugs.com	Monograph
MedlinePlus	a611046
License data	EU EMA: by INN US FDA: abiraterone
Pregnancy category	AU: D US: X (Contraindicated)
Routes of administration	By mouth (tablets)[1]
ATC code	L02BX03 (WHO)
Legal status
Legal status	AU: S4 (Prescription only) CA: ℞-only UK: POM (Prescription only) US: ℞-only
Pharmacokinetic data
Bioavailability	Unknown, but may be 50% at most on empty stomach[3]
Protein binding	Abiraterone: ~99.8% (to albumin and α1-AGp)[3][1][2]
Metabolism	Esterases, CYP3A4, SULT2A1[2]
Metabolites	Abiraterone, others[1][3]
Elimination half-life	Abiraterone: 12–24 hours[1][3]
Excretion	Feces: 88%[1][2] Urine: 5%[1][2]
Identifiers
IUPAC name[show]
CAS Number	154229-18-2  154229-19-3 (abiraterone)
PubChem CID	9821849
IUPHAR/BPS	6745
DrugBank	DB05812
ChemSpider	7997598
UNII	EM5OCB9YJ6
KEGG	D09701
ChEBI	CHEBI:68639
ChEMBL	CHEMBL271227
Chemical and physical data
Formula	C26H33NO2
Molar mass	391.555 g/mol
3D model (JSmol)	Interactive image
Melting point	144 to 145 °C (291 to 293 °F) [4]

Depiction of the Zephyr ® endobronchial valve. Image courtesy of Pulmonx, Inc.

BMS-986169

CAS 1801151-08-5 Related CAS : 1801151-09-6   1801151-08-5
Chemical Formula: C23H27FN2O2
Molecular Weight: 382.4794
Elemental Analysis: C, 72.23; H, 7.12; F, 4.97; N, 7.32; O, 8.37

(R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one

(3R)-3-[(3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl]-1-[(4-methylphenyl)methyl]pyrrolidin-2-one

BMS-986169 is a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate 2B Receptor Negative Allosteric Modulator with Potential in Major Depressive Disorder. BMS-986169 showed high binding affinity for the GluN2B subunit allosteric modulatory site (Ki = 4.03-6.3 nM) and selectively inhibited GluN2B receptor function in Xenopus oocytes expressing human N-methyl-d-aspartate receptor subtypes (IC50 = 24.1 nM). BMS-986169 weakly inhibited human ether-a-go-go-related gene channel activity (IC50 = 28.4 μM) and had negligible activity in an assay panel containing 40 additional pharmacological targets.

Chemical structures of BMS-986169 and the phosphate prodrug BMS-986163.

PAPER

Evolution of a Scale-Up Synthesis to a Potent GluN2B Inhibitor and Its Prodrug

James Kempson*^† , Huiping Zhang^†, Michael K. Y. Wong^†, Jianqing Li^† , Peng Li^†, Dauh-Rurng Wu^†, Richard Rampulla^†, Michael A. Galella^‡, Marta Dabros^‡, Sarah C. Traeger^†, Vetrichelvan Muthalagu^§, Anuradha Gupta^§, Pirama Nayagam Arunachalam^§, and Arvind Mathur^†

^† Discovery Chemistry and Molecular Technologies, Bristol-Myers Squibb Research and Development, Princeton, New Jersey 08540, United States

^‡ Drug Product Science & Technology, Materials Science & Engineering, Bristol-Myers Squibb Research and Development, Princeton, New Jersey 08540, United States

^§ Department of Discovery Synthesis, Biocon Bristol-Myers Squibb Research Center (BBRC), Bangalore 560099, India

Org. Process Res. Dev., Article ASAP

DOI: 10.1021/acs.oprd.8b00120

*E-mail: james.kempson@bms.com.

This paper describes the efficient scale-up synthesis of the potent negative allosteric glutamate N2B (GluN2B) inhibitor 1 (BMS-986169), which relies upon a stereospecific S_N2 alkylation strategy and a robust process for the preparation of its phosphate prodrug 28 (BMS-986163) from parent 1 using POCl₃. A deoxyfluorination reaction employing bis(2-methoxyethyl)aminosulfur trifluoride (Deoxo-Fluor) is also used to stereospecifically introduce a fluorine substituent. The optimized routes have been demonstrated to provide APIs suitable for toxicological studies in vivo.

https://pubs.acs.org/doi/suppl/10.1021/acs.oprd.8b00120/suppl_file/op8b00120_si_001.pdf

PAPER

https://pubs.acs.org/doi/abs/10.1021/acsmedchemlett.8b00080

BMS-986163, a Negative Allosteric Modulator of GluN2B with Potential Utility in Major Depressive Disorder

Lawrence R. Marcin*^† , ET AL

^† Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, Connecticut 06492, United States

^‡ Biocon Bristol-Myers Squibb Research Center, Bangalore, India

^§ Bristol-Myers Squibb Research and Development, 3551 Lawrenceville Road, Princeton, New Jersey 08648, United States

ACS Med. Chem. Lett., 2018, 9 (5), pp 472–477

DOI: 10.1021/acsmedchemlett.8b00080

*Phone 203-677-6701. E-mail: lawrence.marcin@bms.com.

There is a significant unmet medical need for more efficacious and rapidly acting antidepressants. Toward this end, negative allosteric modulators of the N-methyl-d-aspartate receptor subtype GluN2B have demonstrated encouraging therapeutic potential. We report herein the discovery and preclinical profile of a water-soluble intravenous prodrug BMS-986163 (6) and its active parent molecule BMS-986169 (5), which demonstrated high binding affinity for the GluN2B allosteric site (K_i = 4.0 nM) and selective inhibition of GluN2B receptor function (IC₅₀ = 24 nM) in cells. The conversion of prodrug 6 to parent 5 was rapid in vitro and in vivo across preclinical species. After intravenous administration, compounds 5 and 6 have exhibited robust levels of ex vivo GluN2B target engagement in rodents and antidepressant-like activity in mice. No significant off-target activity was observed for 5, 6, or the major circulating metabolites met-1 and met-2. The prodrug BMS-986163 (6) has demonstrated an acceptable safety and toxicology profile and was selected as a preclinical candidate for further evaluation in major depressive disorder.

(S)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-
methylbenzyl)pyrrolidin-2-one (compound 23) and (R)-3-((3S,4S)-3-fluoro-4-(4-
hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrrolidin-2-one (BMS-986169, compound
5)……https://pubs.acs.org/doi/suppl/10.1021/acsmedchemlett.8b00080/suppl_file/ml8b00080_si_001.pdf

Analytical data for BMS-986169 (compound 5): LCMS (C23H27FN2O2, MW 382.2, ESAPI),
observed 383.2 m/z (M+H)+; []D20 = +6.09 (c = 1.15, MeOH); Anal. Calcd for
C23H27FN2O2 (382.21): C, 72.22; H, 7.12; N, 7.32. Found: C, 72.26; H, 7.05; N, 7.31; HRMS
(ESI) Calcd for C23H27N2O2, 383.2118. Found, 383.2129;

13C NMR (126 MHz, chloroformd)
172.4, 155.0, 137.5, 133.0, 132.8, 129.4, 128.6, 128.2, 115.6, 91.6 (d, J=173.5 Hz),
65.0, 54.5 (d, J=25.4 Hz), 48.3, 47.7 (d, J=17.3 Hz), 46.7, 43.6, 31.5, 21.1, 19.2;(500 MHz, chloroform-d) 7.23 – 7.11 (m, 5H), 6.92 (d, J=8.5 Hz, 2H), 6.18 (br. s., 1H),
4.79 – 4.55 (m, 1H), 4.57 – 4.33 (m, 2H), 3.72 (t, J=8.7 Hz, 1H), 3.46 – 3.30 (m, 1H), 3.30 –
3.09 (m, 2H), 2.82 (d, J=8.5 Hz, 1H), 2.73 – 2.56 (m, 2H), 2.49 (d, J=2.5 Hz, 1H), 2.36 (s,
3H), 2.21 – 1.98 (m, 2H), 1.87 (br. s., 2H). The corresponding 1H NMR spectrum for
compound 5 is shown below

1H NMR

PATENT

https://patents.google.com/patent/US9221796B2/und

InventorDalton KingLorin A. Thompson, IIIJianliang ShiSrinivasan ThangathirupathyJayakumar Sankara WarrierImadul IslamJohn E. Macor

Current Assignee Bristol-Myers Squibb Co

https://patents.google.com/patent/WO2015105772A1/und

N-Methyl-D-aspartate (NMDA) receptors are ion channels which are gated by the binding of glutamate, an excitatory neurotransmitter in the central nervous system. They are thought to play a key role in the development of a number of neurological diseases, including depression, neuropathic pain, Alzheimer’s disease, and Parkinson’s disease. Functional NMDA receptors are tetrameric structures primarily composed of two NRl and two NR2 subunits. The NR2 subunit is further subdivided into four individual subtypes: NR2A, NR2B, NR2C, and NR2D, which are differentially distributed throughout the brain. Antagonists or allosteric modulators of NMDA receptors, in particular NR2B subunit-containing channels, have been investigated as therapeutic agents for the treatment of major depressive disorder (G. Sanacora, 2008, Nature Rev. Drug Disc. 7: 426-437).

The NR2B receptor contains additional ligand binding sites in additon to that for glutamate. Non-selective NMDA antagonists such as Ketamine are pore blockers, interfering with the transport of Ca⁺⁺ through the channel. Ketamine has demonstrated rapid and enduring antidepressant properties in human clinical trials as an i.v. drug. Additionally, efficacy was maintained with repeated, intermittent infusions of Ketamine (Zarate et al., 2006, Arch. Gen. Psychiatry 63: 856-864). This class of drugs, though, has limited therapeutic value because of its CNS side effects, including dissociative effects.

An allosteric, non-competitive binding site has also been identified in the N-terminal domain of NR2B. Agents which bind selectively at this site, such as

Traxoprodil, exhibited a sustained antidepressant response and improved side effect profile in human clinical trials as an i.v. drug (Preskorn et al., 2008, J. Clin.

PsychopharmacoL, 28: 631-637, and F. S. Menniti, et al, 1998, CNS Drug Reviews, 4, 4, 307-322). However, development of drugs from this class has been hindered by low bioavailability, poor pharmacokinetics, and lack of selectivity against other pharmacological targets including the hERG ion channel. Blockade of the hERG ion channel can lead to cardiac arrythmias, including the potentially fatal Torsades de pointe, thus selectivity against this channel is critical. Thus, in the treatment of major depressive disorder, there remains an unmet clinical need for the development of effective NR2B-selective negative allosteric modulators which have a favorable tolerability profile.

NR2B receptor antagonists have been disclosed in PCT publication WO 2009/006437.

The invention provides technical advantages, for example, the compounds are novel and are ligands for the NR2B receptor and may be useful for the treatment of various disorders of the central nervous system. Additionally, the compounds provide advantages for pharmaceutical uses, for example, with regard to one or more of their mechanism of action, binding, inhibition efficacy, target selectivity, solubility, safety profiles, or bioavailability.

Synthetic Scheme 1

The l-phenyl/benzyl-3-bromo-pyrrolidinones/piperidinones V may be reacted with (4-oxy-phenyl)cyclic amines VI in the presence of base to produce protected products VII, which may be subjected to cleavage conditions appropriate for the protecting group (PGi) to generate final products I, which may be separated into individual enantiomers/diastereomers I*, as shown in synthetic scheme 2.

Synthetic Scheme 2

I I*

Compounds la may be prepared by condensing l-phenyl/benzyl-3-bromo-pyrroli-dinones/piperidinones V with substituted 4(4-oxyphenyl)piperidines Vllla-c to generate protected intermediates IX, which may be subjected to cleavage conditions appropriate for the protecting group (PGi) to generate final products la, which may be separated into individual enantiomers/diastereomers la*, as shown in synthetic scheme 3.

Synthetic Scheme 3

The 4(4-oxyphenyl)piperidines Vllla-c may be synthesized in turn by a sequence starting with a protected tetrahydropiperidine X, which can be hydroxylated via hydroboration/oxidation to give the protected hydroxypiperidine XI, which may be either directly transformed into the protected fluoropiperidine XII by treatment with DAST or oxidized into the protected 3-oxopiperidine XIII, which may be further transformed into protected 3,3-difluoropiperidines XIV via treatment with DAST. XI, XII, and XIV may be transformed into Villa, Vlllb, and VIIIc, respectively, by employing cleaving conditions appropriate for the protecting group (PG₂), as shown in synthetic scheme 3 a.

S nthetic scheme 3 a

Chiral

Cleavage Individual enantiomers/

G₂P-N diastereomers

separation

conditions

OH 
Villa*

Villa

XI

Chiral

Cleavage Individual enantiomers/

HN

G₂P-N diastereomers

%_\J> PQ separation

PG₁ conditions

R F

F Vlllb*

Vlllb

XII

Chiral

Individual enantiomers/

G₂P- diastereomers separation

Vlllc*

For tetrahydropyridines X which are not commercially available may be synthesized by coupling protected bromophenols XV with protected unsaturated

piperidineboronic acids XVI, as shown in synthetic scheme 4a.

Synthetic scheme 4a:

For tetrahydropyridines X which are not commercially available may be synthesized by adding the anion generated from protected bromophenols XV to a protected 4-piperidinone XVII to yield 4-phenyl-4-piperidinol XVIII, which may be dehydrated under acid conditions to yield the desired X, as shown in synthetic scheme 4b.

Synthetic scheme 4b:

l-Phenyl/benzyl-3-bromo-pyrroli-dinones/piperidinones V may be condensed with isolated individual enantiomers VIIIa-c*, which results in diastereomers 1- phenyl/benzyl-3-bromo-pyrroli-dinones/piperidinones IX*, which may be deprotected and separated to give final products la*, as shown in scheme 5.

Alternatively, the backbone scaffold may be synthesized by condensing 1- phenyl/benzyl-3-bromo-pyrroli-dinones/piperidinones V with hydroxypiperidines Villa to yield the protected 3-fluoropiperidines IXa, which may themselves be converted to the protected 3-fluoropiperidines IXb or oxidized to the ketones XIX, which may be converted to the 3,3-difluoropiperidines Ixc, as shown in scheme 6. The final compounds can then be isolated after the deprotection of IXa-c.

Scheme 6

Example 46, P-1 Example 46, P-2

(S)-3-((3S,4S)-3-Fluoro-4-(4-hydroxyphenyl)piperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one and (R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one.

Example 46, P-3 Example 46, P-4

Step A. (±)-rel-(3S,4S)- 1 -benzyl-4-(4-methoxyphenyl)piperidin-3-ol.

To a suspension of sodium tetrahydroborate (2.7 g, 72 mmol) in THF (200 mL) at 0 °C under a nitrogen atmosphere was added dropwise boron trifluoride etherate (8.8 mL, 70 mmol) and the resulting mixture was stirred for 30 minutes. Then 1-benzyl- 4-(4-methoxyphenyl)-l,2,3,6-tetrahydropyridine (10 g, 36 mmol, from S. Halazy et al WO 97/28140 (8/7/97)) dissolved in 100 mL of tetrahydrofuran was added. The mixture was allowed to warm to rt and stirred for 2 h. The reaction was then quenched by the dropwise addition of 100 mL of water. Next were added

sequentially 100 mL of ethanol, 100 mL of a 10% aqueous sodium hydroxide solution, and 30%> hydrogen peroxide (18 mL, 180 mmol) and the mixture was stirred at reflux temperature overnight. The reaction mixture was then allowed to cool, diluted with saturated aqueous ammonium chloride (200 mL), and extracted with ethyl acetate (500 mL). The organic layer was dried over Na₂S0₄, filtered, and evaporated under reduced pressure to give (±)-rel-(3S,4S)- 1 -benzyl-4-(4-methoxyphenyl)piperidin-3-ol (8.5 g, 24.6 mmol, 69%> yield) which was used without further purification. LCMS (Method K) RT 1.99 min; m/z 298.0 (M+H⁺).

Step B. (±)-re -(3S,4S)-4-(4-methoxyphenyl)piperidin-3-ol.

To a solution of (±)-re/-(35′,45)-l-benzyl-4-(4-methoxyphenyl)piperidin-3-ol (9 g, 30 mmol) in methanol (150 mL) was added 10 % Pd/C (4.8 g) and the reaction mixture was stirred overnight under a hydrogen atmosphere. The catalyst was then removed by filtration through Celite and the solvent was evaporated under reduced pressure to give (±)-re/-(3S,4S)-4-(4-methoxyphenyl)piperidin-3-ol (5.1 g, 24.6 mmol, 81% yield) which was used without further purification. 1H NMR (400 MHz, DMSO-de) δ ppm 7.10 – 7.15 (m, 2 H) 6.80 – 6.86 (m, 2 H) 4.30 (d, J=5.27 Hz, 1 H) 3.37 – 3.43 (m, 1 H) 3.04 (dd, J=11.58, 4.36 Hz, 1 H) 2.86 (d, J=12.17 Hz, 1 H) 2.43 (td, J=12.09, 2.67 Hz, 1 H) 2.22 – 2.35 (m, 2 H) 1.57 – 1.63 (m, 1 H) 1.43 – 1.54 (m, 1 H).

To a solution of (±)-re/-(3S,4S)-4-(4-methoxyphenyl)piperidin-3-ol (4.5 g, 21.7 mmol) in DCM (150 mL) at -10°C under nitrogen was added a 1 M solution of boron tribromide in DCM (109 mL, 109 mmol). The reaction mixture was allowed to warm to rt, stired for 2 h, and then rechilled to 0 °C and quenched by the addition of a saturated aqueous sodium bicarbonate solution (300 mL). The aqueous layer was washed with 250 mL of DCM and then to it was added 200 mL 10% aqueous NaOH, followed by 9.5 g (43.5 mmol) of di-t-butyl dicarbonate and the resulting mixture was stirred for an additional 2 h. The mixture was then extracted with 200 mL ethyl acetate and the organic layer was separated, dried over Na₂S0₄,filtered, and evaporated under reduced pressure to (±)-re/-(35′,45)-tert-butyl 4-(4-(tert-butoxycarbonyloxy)phenyl)-3-hydroxypiperidine-l-carboxylate (6.5 g, 12 mmol, 56 % yield) which was used without further purification. LCMS (Method K) RT 2.33 min, m/z 282 (M+H⁺ -2 t-butyl), 370; 1H NMR (400 MHz, DMSO-d₆) δ ppm 7.27 (d, J=8.66 Hz, 2 H) 7.08 (d, J=8.66 Hz, 2 H) 4.85 (d, J=5.65 Hz, 1 H) 4.13 (d, J=8.41 Hz, 1 H) 3.97 (d, J=10.48 Hz, 1 H) 3.45 (tt, J=10.27, 5.19 Hz, 1 H) 1.67 (d, J=3.39 Hz, 1 H) 1.50 – 1.59 (m, 1 H) 1.49 (s, 11 H).

Step D. (±)-re/-(35′,45)-tert-butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine-l-carboxylate.

To a solution of (±)-re/-(35′,45)-tert-butyl 4-(4-(tert-butoxycarbonyloxy)phenyl)-3-hydroxypiperidine-l-carboxylate (6.5 g, 16.5 mmol) in 100 mL of methanol was added 11.42 g of potassium carbonate (83 mmol) and the reaction mixture was stirred at rt for 5 h. The organic solvent was removed under reduced pressure and the residue was partitioned between IN HC1 (300 mL) and ethyl acetate (300 mL). The layers were separated and the organic layer was dried over Na₂S0₄ and evaporated under reduced pressure to give (±)-re/-(35′,45)-tert-butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine-l-carboxylate (5 g, 15 mmol, 92 % yield) which was used without further purification. LCMS (method F) RT 1.85 min, m/z 238 (M+H⁺ – 1-butyl), 279 (M+H⁺ – t-butyl+CH₃CN), 1H NMR (400 MHz, DMSO-d₆) δ ppm 7.01 (d, J=8.53 Hz, 2 H) 6.66 (d, J=8.53 Hz, 2 H) 4.70 (d, J=5.02 Hz, 1 H) 4.09 (br. s., 1 H) 3.94 (d, J=11.55 Hz, 1 H) 3.35 – 3.41 (m, 1 H) 2.66 – 2.77 (m, 1 H) 2.29 – 2.39 (m, 1 H) 1.63 (dd, J=13.30, 3.26 Hz, 1 H) 1.44 – 1.52 (m, 1 H) 1.42 (s, 9 H).

Step E. (3S,4S)-tert-Butyl 3 -hydroxy-4-(4-hydroxyphenyl)piperidine-l -carboxylate and (3R, -tert-butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine-l-carboxylate.

E-1 E-2

(±)-rel-(3S,4S)-tert-Butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine- 1 -carboxylate (5 g, 17 mmol, from step D) was subjected to chiral SFC separation (method C-5) to yield enantiomers E-1 (1.9 g, 6.48 mmol, 38.0 % yield) and E-2 (2.4 g, 8.18 mmol, 48.0 % yield). Data for E-1 : chiral HPLC (method A5 ) retention time 3.42 min. Data for E-2: chiral HPLC (method A5) retention time 4.2 min.

Step F. (3R,4R)-tert-Butyl 4-(4-(benzyloxy)phenyl)-3-hydroxypiperidine-l-carboxylate.

A mixture of (3R,4R)-tert-butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine-l-carboxylate (620 mg, 2.1 mmol, E-2 from step E), potassium carbonate (584 mg, 4.2 mmol), and benzyl bromide (0.25 mL, 2.1 mmol) in DMF (5 mL) was stirred at rt for 16 h. The solvent was removed by evaporation and the residue was treated with 50 mL of water. The aqueous mixture was then extracted 4 times with 50 mL of chloroform. The combined organic phases were dried over anhydous Na₂S0₄, filtered, and evaporated to yield 750 mg of (3R,4R)-tert-butyl 4-(4-(benzyloxy)phenyl)-3-hydroxypiperidine-l -carboxylate which was used without further purification. LCMS (method F) RT 2.28 min, m/z = 310 (M+H⁺ – t-butyl -water), 328 (M+H⁺ -t-butyl).

Step G. (3i?,4i?)-4-(4-(Benzyloxy)phenyl)piperidin-3-ol hydrochloride.

A mixture of (3R,4R)-tert-butyl 4-(4-(benzyloxy)phenyl)-3-hydroxypiperidine-1-carboxylate (750 mg, 2 mmol), dioxane (4 mL) and 4.9 mL of 4 M HCI in dioxane was stirred at rt for 2h. The reaction was then evaporated to dryness to yield 550 mg of (3i?,4i?)-4-(4-(Benzyloxy)phenyl)piperidin-3-ol hydrochloride which was used without further purification. LCMS (method J) RT 0.70 min, m/z 284 (M+H⁺).

Step H. 3-((3i?,4i?)-4-(4-(Benzyloxy)phenyl)-3-hydroxypiperidin-l -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one .

A mixture of 3-bromo-l-(4-methylbenzyl)pyrrolidin-2-one (Intermediate 2, 220 mg, 0.82 mmol), (3i?,4i?)-4-(4-(benzyloxy)phenyl)piperidin-3-ol hydrochloride (262 mg, 0.82 mmol, from step G) and triethylamine (11 mL, 8.2 mmol) was stirred at 60 °C for lh, 80 °C for 1 h, 100 °C for 1 h and 120 °C for 1 h. The reaction mixture was then allowed to cool, diluted with 40 mL of water and extracted four times with 50 mL of chloroform. The combined organic layers were washed with 60 mL brine, dried over anhydrous sodium sulfate, filtered, and evaporated to yield 382 mg of 3-((3 ?,4i?)-4-(4-(benzyloxy)phenyl)-3-hydroxypiperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one which was used without further purification. LCMS (method J) (main component of a mixture) RT 2.23 min, m/z 471 (M+H⁺).

Step I. 3-((3R, 4R)-4-(4-(Benzyloxy)phenyl)-3-fluoropiperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one .

A solution of 3-(-4-(4-(benzyloxy)phenyl)-3-hydroxypiperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one (382 mg, 0.81 mmol) in DCM (5 mL) cooled to 0 °C was treated dropwise with DAST (0.32 mL, 2.4 mmol) over 3 min. The reaction mixture was then allowed to warm to rt and was stirred for 2 h. The reaction was then quenched with 50 mL of 10% aqueous sodium bicarbonate solution and extracted 4 times with 40 mL of DCM. The combined organic layers were washed with 50 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to yield 382 mg of 3-((3i?,4i?)-4-(4-(benzyloxy)phenyl)-3-fluoropiperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one as a mixture of two diastereomers and rearrangement products which was used without further purification. LCMS (method J) (main component of a mixture) RT 0.9 min, m/z 473 (M+H⁺).

Step J. 3-((3i?,4i?)-3-Fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)-l -(4-methylbenzyl)pyrrolidin-2-one .

A mixture of 3-((Ji?,4i?)-(4-(4-(benzyloxy)phenyl)-3-fluoropiperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one (382 mg, 0.81 mmol) and methanol (4 mL) was flushed with nitrogen, followed by the addition of 172 mg of 10% Pd/C. Then the mixture was stirred at rt overnight under 25-99 psi hydrogen pressure. The reaction was then transferred to a 100 mL autoclave and stirred at 7 kg/cm² hydrogen pressure for 4 days. The catalyst was removed by filtration through Celite and the solvent was evaporated off. The crude product was subjected to HPLC purification (method B) to yield 77.3 mg 3-((Ji?,4i?)-3-fluoro-4-(4-hydroxyphenyl)-piperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one (diastereomeric pair) LCMS (method Q) RT 1.15 min, m/z 383.0 (M+H⁺).

Step K. (5)-3-((3i?,4i?)-3-Fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl>

methylbenzyl)pyrrolidin-2-one and (i?)-3-((3i?,4i?)-3-fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one.

The diastereomeric mixture from step J was separated by SFC method C-7 to yield homochiral Examples 46 P-l (29.3 mg) and P-2 (32.8 mg). Data for P-l (S)-3-((3R, 4R)-3 -fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one: LCMS (method F) RT 2.10 min, m/z 383.2 (M+H⁺), 405.2 (M+Na⁺); HPLC (method B) RT 8.24 min (98.8% AP); HPLC (method C) RT 6.52 min (99.1% AP); Chiral HPLC (method C-6) RT 4.1 min; 1H NMR (400 MHz, methanol-d₄) δ ppm 1.76 – 1.86 (m, 2 H) 2.07 (d, J=8.53 Hz, 1 H) 2.13 – 2.21 (m, 1 H) 2.34 (s, 3 H) 2.43 (s, 0 H) 2.55 – 2.60 (m, 1 H) 2.65 – 2.70 (m, 1 H) 2.75 (br. s., 1 H) 3.20 – 3.30 (m, 2 H) 3.38 – 3.45 (m, 1 H) 3.70 (t, J=8.78 Hz, 1 H) 4.44 (t, J=79.81 Hz, 3 H) 4.63 – 4.71 (m, 1 H) 6.70 – 6.80 (m, 2 H) 7.07 – 7.15 (m, 2 H) 7.07 – 7.12 (m, 1 H) 7.13 – 7.22 (m, 4 H); ¹⁹F NMR δ ppm -184.171. Data for P-2: (R)-3-((3R,4R)-3-fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one: LCMS (method F) RT 2.10 min, m/z 383.2 (M+H⁺), 405.2 (M+Na⁺); HPLC (method B) RT 8.29 min (99.7% AP); HPLC (method C) RT 6.52 min (99.8% AP); Chiral HPLC (method C-6) RT 6.92 min; 1H NMR (400 MHz, methanol-d₄) δ ppm 1.80 – 1.90 (m, 2 H) 2.07 (d, J=8.03 Hz, 1 H) 2.19 (s, 1 H) 2.34 (s, 3 H) 2.41 – 2.48 (m, 1 H) 2.66 (d, J=4.52 Hz, 2 H) 2.95 – 3.03 (m, 1 H) 3.10 – 3.18 (m, 1 H) 3.20 – 3.30 (m, 2 H) 3.68 – 3.78 (m, 1 H) 4.38 (s, 1 H) 4.51 (d, J=14.56 Hz, 2 H) 6.70 – 6.80 (m, 2 H) 7.05 – 7.13 (m, 2 H) 7.13 – 7.22 (m, 4 H); ¹⁹F NMR δ ppm -184.311.

(3S,4S)-tert-Butyl 3-fluoro-4-(4-hydroxyphenyl)piperidine-l-carboxylate.

To a solution of (3S,4S)-tert-butyl 3-hydroxy-4-(4-hydroxyphenyl)piperidine-l-carboxylate (400 mg, 1.36 mmol, the first eluting enantiomer E-l from step E) in DCM (5 mL) cooled to 0 °C was added dropwise DAST (0.54 mL, 4.1 mmol) over 10 min. The mixture was allowed to warm up to rt and was stirred for 2h. The reaction was slowly quenched with 50 mL of a 10%> aqueous sodium bicarbonate solution and extracted four times with 50 mL of DCM. The combined organic layerss were washed with 75 mL of brine, dried, and concentrated under vacuum to yield 390 mg of {3S,4S)-tert-bvXy\ 3-fluoro-4-(4-hydroxyphenyl)piperidine-l-

carboxylate which was used without further purification. LCMS (Method Q) RT 0.92 min, m z 240.1(M+H⁺).

Step M. 4-((3S’,4S)-3-Fluoropi ridin-4-yl)phenol hydrochloride.

A mixture of (3S,4S)-tert-butyl 3-fluoro-4-(4-hydroxyphenyl)piperidine-l-carboxylate (390 mg, 1.3 mmol) and 4M HC1 in dioxane (3.3 mL, 13.2 mmol) in dioxane (4 mL) was stirred at rt for 2 hr. It was then concentrated to dryness, washed with 10 mL of 5% DCM/diethyl ether mixture and the solid was isolated by filtration. Yield: 260 mg of 4-((J£4S)-3-fluoropiperidin-4-yl)phenol hydrochloride; LCMS

(method Q) RT 0.46 min, mz 196.1(M+H⁺) 1H NMR (400 MHz, DMSO-d₆) δ = 9.57 (br. s., 4 H), 8.92 – 8.68 (m, 1 H), 7.14 (d, J= 8.5 Hz, 1 H), 7.06 (d, J= 8.5 Hz, 2 H), 6.82 – 6.73 (m, 2 H), 5.07 – 4.85 (m, 1 H), 3.77 – 3.36 (m, 9 H), 3.32 – 3.22 (m, 2 H), 3.13 – 2.85 (m, 5 H), 2.06 – 1.88 (m, H).

Step N. 3-((3S,4S)-3-Fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one.

A mixture of 3-bromo-l-(4-methylbenzyl)pyrrolidin-2-one (200 mg, 0.75 mmol), triethylamine (0.52 mL, 3.7 mmol) and 4-((3S,4S)-3-fluoropiperidin-4-yl)phenol hydrochloride (173 mg, 0.75 mmol) in DMF (3 mL) was heated to 120 °C in a microwave reactor for 1.5 h. The mixture was allowed to cool and was then mixed with 60 mL water and extracted 5 times with 40 mL of DCM. The combined organic extracts were washed with 80 mL of brine, dried over anhydrous sodium sulfate, filtered, and evaporated to give 265 mg of 3-((3 4S)-3-fluoro-4-(4-hydroxy-phenyl)piperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one as a mixture of 2 diastereoisomers. LCMS (method P) RT 0.92 min m/z 383.4 (M+H⁺).

Step O. (5)-3-((3_lS,45)-3-Fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one and (i?)-3-((35^,,45)-3-fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one.

A portion of the diasteromer mixture from step N (130 mg) was subjected to chiral purification via SFC (method C-7) to give homochiral Examples 46 P-3 (37.7 mg) and P-4 (60.7 mg). Data for P-3 (S)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin- 1 -yl)- 1 -(4-methylbenzyl)pyrrolidin-2-one: LCMS (Method F) RT = 2.10 min, m/z 383.2 (M+H⁺); HPLC (Method C) RT 6.54 min, (Method D) RT 8.20 min; chiral HPLC (method C-6) RT 3.42 min;1H NMR (400 MHz, methanol-d₄) δ ppm 1.76 – 1.86 (m, 2 H) 2.06 (d, J=8.53 Hz, 1 H) 2.10 – 2.21 (m, 1 H) 2.34 (s, 3 H) 2.40 – 2.48 (m, 1 H) 2.53 – 2.60 (m, 1 H) 2.61 – 2.70 (m, 2 H) 2.95 -3.01 (m, 1 H) 3.01 (s, 2 H) 3.10 – 3.16 (m, 1 H) 3.18 – 3.28 (m, 2 H) 3.72 (s, 1 H) 4.35 – 4.41 (m, 1 H) 4.46 – 4.70 (m, 2 H) 6.72 – 6.80 (m, 2 H) 7.05 – 7.23 (m, 6 H). Data for P-4 (R)-3-((3S,4S)-3-fiuoro-4-(4-hydroxyphenyl)piperidin-l-yl)-l-(4-methylbenzyl)pyrrolidin-2-one: LCMS (Method F) RT 2.11 min, m/z 383.2 (M+H⁺);; HPLC (Method C) RT 6.50 min, (Method D) RT 8.21 min; chiral HPLC (method C-6) RT 6.31 min; 1H NMR (400 MHz, methanol-d₄) δ ppm 1.81 (dd, J=7.28, 2.76 Hz, 2 H) 2.06 (d, J=9.04 Hz, 2 H) 2.33 (s, 3 H) 2.43 (s, 1 H) 2.55 (br s, 1 H) 2.66 (d, J=40.16 Hz, 2 H) 2.75 – 2.80 (m, 1 H) 2.96 – 3.10 (m, 2 H) 3.20 – 3.28 (m, 2 H) 3.41 (d, J=5.52 Hz, 1 H) 3.66 – 3.75 (m, 1 H) 4.31 – 4.41 (m, 1 H) 4.46 – 4.71 (m, 2 H) 6.76 (d, J=8.53 Hz, 2 H) 7.05 – 7.23 (m, 6 H).

Example 48 (Peak 1, Peak 2, Peak 3, Peak 4)

(S)-l-(4-fluorobenzyl)-3-((3S,4S)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l- yl)pyrrolidin-2-one, (S)-l-(4-fluorobenzyl)-3-((3R,4R)-3-hydroxy-4-(4- methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one, (R)-l-(4-fluorobenzyl)-3-((3S,4S)-3- hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one, and (R)-l-(4- fluorobenzyl)-3-((3RAR)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2- one

Step A. (±)-re/-l-(4-Fluorobenzyl)-3-((3S,4S)-3-hydroxy-4-(4- methoxyphenyl)piperidin- 1 -yl)pyrrolidin-2-one.

To a solution of 3-bromo-l-(4-fluorobenzyl)pyrrolidin-2-one (Intermediate 1, 300 mg, 1.1 mmol) and trans-4-(4-methoxyphenyl)piperidin-3-ol (from Example 46, step B, 240 mg, 1.16 mmol) in acetonitrile (10 mL) was added triethylamine (560 mg, 5.5 mmol) and the mixture was heated at 120 °C in a microwave reactor for 1 h. The reaction mixture was then diluted with water and extracted with 100 mL of ethyl acetate. The organic layer was dried over Na₂S0₄, filtered, and evaporated under reduced pressure to give (±)-re/-l-(4-fluorobenzyl)-3-((3S,4S)-3-hydroxy-4-(4- methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one (450 mg, 0.7 mmol) as a mixture of four diastereomers which was used without further purification. LCMS (Method S) RT 1.89 min, m/z 399.1 (M+H⁺).

Step B. (±)-re/-3-((3_lS,45)-3-Fluoro-4-(4-methoxyphenyl)piperidin- 1 -yl)- 1 -(4-fluorobenzyl)pyrrolidin-2-one.

To a solution of l-(4-fluorobenzyl)-3-(trans-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one from step B (2.5 g, 6.3 mmol) in 50 mL DCM was added DAST (4.1 mL, 31 mmol) and the reaction was stirred at ambient temperature for 1 h. The reaction was then quenched with a sat.bicarbonate solution (200 mL) and the mixture was extracted with 200 mL of DCM. The organic layer was dried over Na₂S0₄, filtered, and evaporated under reduced pressure. The residue was purified via silica gel chromatography eluting with 28% ethyl acetate in hexane to give (±)-re/-3-((35′,45)-3-fluoro-4-(4-methoxyphenyl)piperidin-l-yl)-l-(4-fluorobenzyl)pyrrolidin-2-one (900 mg, 1.6 mmol) as a mixture of four

diastereomers. LCMS (method P) RT 0.89 min, m/z 401.2 (M+H⁺).

Step C. 3-((3_lS,45)-3-Fluoro-4-(4-hydroxyphenyl)piperidin-l-yl)-l-(4-fluorobenzyl)pyrrolidin-2-one.

To a solution of (tra/?5-3-fluoro-4-(4-methoxyphenyl)piperidin-l-yl)-l-(4-fluorobenzyl)pyrrolidin-2-one (700 mg, 1.75 mmol) in 50 mL of DCM at 0 °C was added BBr₃(0.3 mL, 3.5 mmol). The reaction mixture was allowed to warm up to room temperature over 1 h. The mixture was then diluted with a sat. bicarbonate solution and extracted with 200 mL of DCM. The organic layer was dried over Na₂S0₄, filtered, and evaporated under reduced pressure.

The residue was purified by preparative HPLC(method A) to yield 120 mg of 3-((35′,45)-3-fluoro-4-(4-hydroxyphenyl)piperidin-l-yl)-l-(4-fluorobenzyl)pyrrolidin-2-one as a mixture of four diastereomers. LCMS (method N) RT 1.45 min, m/z 387.0 (M+H⁺).

Step D. (S)- 1 -(4-fluorobenzyl)-3-((35′,45)-3-hydroxy-4-(4-methoxyphenyl)piperidin-1 -yl)pyrrolidin-2-one, (S)- 1 -(4-fluorobenzyl)-3-((3i?,4i?)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one, (i?)-l-(4-fluorobenzyl)-3-((35′,45)-3-hydroxy-4-(4-methoxyphenyl)piperidin- 1 -yl)pyrrolidin-2-one, and (R)- 1 -(4-fluorobenzyl)-3-((3i?,4i?)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one.

The diastereomeric mixture from step C was separated via chiral SFC (method F) into the 4 homochiral diastereomers, Example 48 P-1, P-2, P-3, and P-4. Data for P-1 (S)- 1 -(4-fluorobenzyl)-3-((35′,45)-3-hydroxy-4-(4-methoxyphenyl)piperidin- 1 -yl)pyrrolidin-2-one: Chiral SFC (Method F) RT 3.32 min, 100% AP; HPLC (Method A) RT 6.53 min, 96.0%AP, (Method B) RT 6.7 min, 96.3 %AP; LCMS (Method F) RT 2.02 min, m/z 387.0 (M+H⁺); 1H NMR (400 MHz, methanol-^) δ ppm 7.32 (dd, J=8.78, 5.27 Hz, 2 H) 7.07 – 7.14 (m, 4 H) 6.76 (d, J=8.53 Hz, 2 H) 4.41 – 4.56 (m, 2 H) 3.74 (t, J=8.78 Hz, 1 H) 3.23 – 3.31 (m, 2 H) 3.10 – 3.17 (m, 1 H) 3.01 (d, J=11.04 Hz, 1 H) 2.88 (d, J=7.03 Hz, 1 H) 2.66 (td, J=10.04, 4.52 Hz, 1 H) 2.57 (dd, J=10.54, 6.53 Hz, 1 H) 2.40 – 2.49 (m, 1 H) 2.16 – 2.25 (m, 1 H) 2.03 – 2.13 (m, 1 H) 1.80 -1.88 (m, 2 H). Data for P-2 (5)-l-(4-fluorobenzyl)-3-((3i?,4i?)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one: Chiral SFC (Method F) RT 4.15 min, 99.7% AP; HPLC (Method A) RT 6.52 min, 98.1%AP, (Method B) RT 6.92 min, 98.6 %AP; LCMS (Method F) RT 2.03 min, m/z 387.0 (M+H⁺);1H NMR (400 MHz, methanol-^) δ ppm 7.29 – 7.34 (m, 2 H) 7.07 – 7.14 (m, 4 H) 6.76 (d, J=9.04 Hz, 2 H) 3.71 (t, J=8.78 Hz, 1 H) 3.39 – 3.45 (m, 1 H) 3.24 – 3.31 (m, 2 H) 2.74 -2.80 (m, 1 H) 2.64 – 2.72 (m, 1 H) 2.57 (dd, J=10.54, 6.02 Hz, 1 H) 2.43 (td, J=10.04, 5.02 Hz, 1 H) 2.15 – 2.25 (m, 1 H) 2.05 – 2.14 (m, 1 H) 1.77 – 1.85 (m, 2 H). Data for P-3 (i?)-l-(4-fluorobenzyl)-3-((35′,45)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one: Chiral SFC (Method F) RT 4.56 min, 97.4% AP; HPLC (Method A) RT 6.53 min, 96.0%AP, (Method B) RT 6.94 min, 96.4 %AP; LCMS (Method F) RT 2.02 min, m/z 387.0 (M+H⁺); 1H NMR (400 MHz, methanol-^) δ ppm 7.29 -7.34 (m, 2 H) 7.07 – 7.13 (m, 4 H) 6.74 – 6.78 (m, 2 H) 4.40 – 4.55 (m, 2 H) 3.71 (t, J=9.04 Hz, 1 H) 3.38 – 3.45 (m, 1 H) 3.23 – 3.31 (m, 2 H) 2.76 (br. s., 1 H) 2.64 -2.72 (m, 1 H) 2.57 (dd, J=10.54, 6.02 Hz, 1 H) 2.43 (td, J=10.04, 5.02 Hz, 1 H) 2.16 – 2.25 (m, 1 H) 2.05 – 2.14 (m, 1 H) 1.77 – 1.87 (m, 2 H). Data for P-4 (R)-\-(4- fluorobenzyl)-3-((3i?,4i?)-3-hydroxy-4-(4-methoxyphenyl)piperidin-l-yl)pyrrolidin-2-one: Chiral SFC (Method F) RT 5.57 min, 99.9% AP; HPLC (Method A) RT 6.55 min, 99.9%AP, (Method B) RT 6.90 min, 99.9 %AP; LCMS (Method F) RT 2.03 min, m/z 387.0 (M+H⁺); 1H NMR (400 MHz, methanol-^) δ ppm 7.32 (dd, J=8.78, 5.27 Hz, 2 H) 7.07 – 7.14 (m, 4 H) 6.76 (d, J=8.53 Hz, 2 H) 4.41 – 4.56 (m, 3 H) 3.74 (t, J=8.53 Hz, 1 H) 3.24 – 3.32 (m, 2 H) 3.10 – 3.17 (m, 1 H) 2.66 (td, J=9.91, 4.77 Hz, 1 H) 2.57 (dd, J=10.54, 6.53 Hz, 1 H) 2.41 – 2.49 (m, 1 H) 2.16 – 2.24 (m, 1 H) 2.04 – 2.12 (m, 1 H) 1.80 – 1.88 (m, 2 H).

ADDITIONAL INFORMATION

Intravenous administration of BMS-986169 or BMS-986163 dose-dependently increased GluN2B receptor occupancy and inhibited in vivo [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine ([3H]MK-801) binding, confirming target engagement and effective cleavage of the prodrug. BMS-986169 reduced immobility in the mouse forced swim test, an effect similar to intravenous ketamine treatment. Decreased novelty suppressed feeding latency, and increased ex vivo hippocampal long-term potentiation was also seen 24 hours after acute BMS-986163 or BMS-986169 administration. BMS-986169 did not produce ketamine-like hyperlocomotion or abnormal behaviors in mice or cynomolgus monkeys but did produce a transient working memory impairment in monkeys that was closely related to plasma exposure. Finally, BMS-986163 produced robust changes in the quantitative electroencephalogram power band distribution, a translational measure that can be used to assess pharmacodynamic activity in healthy humans. Due to the poor aqueous solubility of BMS-986169, BMS-986163 was selected as the lead GluN2B NAM candidate for further evaluation as a novel intravenous agent for TRD.

REFERENCES

1: Bristow LJ, Gulia J, Weed MR, Srikumar BN, Li YW, Graef JD, Naidu PS, Sanmathi
C, Aher J, Bastia T, Paschapur M, Kalidindi N, Kumar KV, Molski T, Pieschl R,
Fernandes A, Brown JM, Sivarao DV, Newberry K, Bookbinder M, Polino J, Keavy D,
Newton A, Shields E, Simmermacher J, Kempson J, Li J, Zhang H, Mathur A, Kallem
RR, Sinha M, Ramarao M, Vikramadithyan RK, Thangathirupathy S, Warrier J, Islam
I, Bronson JJ, Olson RE, Macor JE, Albright CF, King D, Thompson LA, Marcin LR,
Sinz M. Preclinical Characterization of
(R)-3-((3S,4S)-3-fluoro-4-(4-hydroxyphenyl)piperidin-1-yl)-1-(4-methylbenzyl)pyrr
olidin-2-one (BMS-986169), a Novel, Intravenous, Glutamate N-Methyl-d-Aspartate
2B Receptor Negative Allosteric Modulator with Potential in Major Depressive
Disorder. J Pharmacol Exp Ther. 2017 Dec;363(3):377-393. doi:
10.1124/jpet.117.242784. Epub 2017 Sep 27. PubMed PMID: 28954811.

2. BMS-986163, a Negative Allosteric Modulator of GluN2B with Potential Utility in Major Depressive Disorder
Lawrence R. Marcin, Jayakumar Warrier, Srinivasan Thangathirupathy, Jianliang Shi, George N. Karageorge, Bradley C. Pearce, Alicia Ng, Hyunsoo Park, James Kempson, Jianqing Li, Huiping Zhang, Arvind Mathur, Aliphedi B. Reddy, G. Nagaraju, Gopikishan Tonukunuru, Grandhi V. R. K. M. Gupta, Manjunatha Kamble, Raju Mannoori, Srinivas Cheruku, Srinivas Jogi, Jyoti Gulia, Tanmaya Bastia, Charulatha Sanmathi, Jayant Aher, Rajareddy Kallem, Bettadapura N. Srikumar, Kumar Kuchibhotla Vijaya, Pattipati S. Naidu, Mahesh Paschapur, Narasimharaju Kalidindi, Reeba Vikramadithyan, Manjunath Ramarao, Rex Denton, Thaddeus Molski, Eric Shields, Murali Subramanian, Xiaoliang Zhuo, Michelle Nophsker, Jean Simmermacher, Michael Sinz, Charlie Albright, Linda J. Bristow, Imadul Islam, Joanne J. Bronson, Richard E. Olson, Dalton King, Lorin A. Thompson, and John E. Macor
Publication Date (Web): April 13, 2018 (Letter)
DOI: 10.1021/acsmedchemlett.8b00080

//////////////////BMS-986169, BMS-986169, BMS 986169, BMS986169

 O=C1N(CC2=CC=C(C)C=C2)CC[C@H]1N3C[C@@H](F)[C@H](C4=CC=C(O)C=C4)CC3

CH4630808, CH-4630808, NA-808

(2S)-2-[(E,2S)-1-[[(1S)-2-(4-but-2-ynoxyphenyl)-1-carboxyethyl]amino]-1,11-dioxooctadec-3-en-2-yl]-2-hydroxybutanedioic acid

Cas 827034-92-4  DOUBLE BOND E, SP ROT (-)

CAS 744208-75-1  E Z NOT DEFINED

• D-erythro-Pentonic acid, 5-[[(1S)-2-[4-(2-butynyloxy)phenyl]-1-carboxyethyl]amino]-3-C-carboxy-2,4,5-trideoxy-5-C-oxo-4-[(1E)-9-oxo-1-hexadecenyl]- (9CI)
• 5-[[(1S)-2-[4-(2-Butyn-1-yloxy)phenyl]-1-carboxyethyl]amino]-3-C-carboxy-2,4,5-trideoxy-5-C-oxo-4-[(1E)-9-oxo-1-hexadecen-1-yl]-D-erythro-pentonic acid
• D-erythro-Pentonic acid, 5-[[(1S)-2-[4-(2-butyn-1-yloxy)phenyl]-1-carboxyethyl]amino]-3-C-carboxy-2,4,5-trideoxy-5-C-oxo-4-[(1E)-9-oxo-1-hexadecen-1-yl]-

Chugai Pharmaceutical (Originator)

Trisodium Der ,CAS 1799542-36-1,  SP ROT (-), MW 709.7097, MF C35 H46 N O10 . 3 Na, Trisodium (2S)-2-[(2S,3E)-1-([(1S)-2-[4-(but-2-yn-1-yloxy)phenyl]-1-carboxylatoethyl]amino)-1,11-dioxooctadec-3-en-2-yl]-2-hydroxybutanedioate

SIMILAR

PAPER

https://www.sciencedirect.com/science/article/pii/S0960894X12013741

Bioorganic & Medicinal Chemistry Letters

Scheme 3. Reagents and conditions: (a) TBDPSCl, imidazole, DMF, rt; (b) n-BuLi, (CH2O)n, THF; (c) Red-Al, 0 C, then I2, THF 40 C; (d) DHP, PPTS, DCM, rt; (e) n-BuLi, (CH2O)n, THF, 78 C to 0 C; (f) TBDPSCl, imidazole, DMF, rt; (g) PPTS, EtOH. 28.6% over 7 steps; (h) L-(+)-DET, Ti(Oi-Pr)4, TBHP, DCM, 97%, >95% ee; (i) Terminal alkyne 7 in Scheme 2, Cp2ZrClH, MeMgCl, CuI, THF, 20 C, 91% yield⁄ ; (j) 2,2-dimethoxypropane, PPTS, DCM, 85% yield⁄ ; (k) TBAF, AcOH, THF, 89% yield⁄ ; (l) oxalyl chloride, DMSO, triethylamine, DCM, 78 C; (m) NaClO2, NaH2PO4, 2-methyl-2-butene, t-BuOH-H2O; (n) N,N-dimethylformamide di-tert-butyl acetal, 58% yield in 3 steps⁄ ; (o) 80% AcOH, THF, rt, 90% yield⁄ ; (p) Jones reagent, aqueous acetone, 10 C, 80% yield⁄ ; (q) the corresponding amine, HATU, Hunig base, 85% yield⁄ ; (r) TFA, anisole, DCM, 90% yield⁄ ; (s) H2-Pd/C, EtOH, 80%; (t) NaBH4, THF, MeOH, 93% yield. ⁄ yields when n = 5 and R1 = n-C7H15.

Paper

Development of a Kilogram-Scale Synthesis of a Novel Anti-HCV Agent, CH4930808

CH4630808 corrected

Tsuyoshi Haneishi^*† , Yoshiaki Kato^‡, Hiroshi Fukuda^†, Tomoyuki Shimamura^‡, Takemi Tanokura^‡, Akira Hiraide^‡, Kaichiro Koyama^‡, Masato Fudesaka^‡, Kenji Maeda^‡, Nobuyuki Nakata^‡, Masahiro Nagase^‡, Takahiko Yabuzaki^‡, Hiroaki Takao^‡, Masaharu Kigawa^‡, Hitoshi Shimizu^‡, and Makoto Shimizu^§

^† Research Division, Chugai Pharmaceutical Co., Ltd., 1-135 Komakado, Gotemba, Shizuoka 412-8513, Japan

^‡ Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-5-1 Ukima, Kita-ku, Tokyo 115-8543, Japan

^§ Department of Chemistry for Materials, Graduate School of Engineering, Mie University, Tsu, Mie 514-8507, Japan

Org. Process Res. Dev., 2018, 22 (2), pp 236–240

DOI: 10.1021/acs.oprd.7b00383

*E-mail: haneishitys@chugai-pharm.co.jp. Tel.: +81-550-87-9102. Fax: +81-550-87-5326.

Abstract

Herein, we report the kilogram-scale synthesis of CH4930808 (1) CH 4630808 CORRECTED, a novel anti-hepatitis C virus agent. While pursuing improved productivity using many through-process strategies, we conducted scrupulous impurity control. Finally, we successfully developed a practical and scalable process for the synthesis of (1·1.5Na·2.5H₂O), by which we prepared 3.28 kg of the active pharmaceutical ingredient for clinical studies

https://pubs.acs.org/doi/suppl/10.1021/acs.oprd.7b00383/suppl_file/op7b00383_si_001.pdf

1H-NMR and 13C-NMR spectra of compound 5·HCl S 3– S 4

1H-NMR spectra of compound 1·1.5 Na·2.5 H2O S 5

13C-NMR spectra compound 1·1.5 Na·2.5 H2O S 6

1H-COSY spectra of compound 1·1.5 Na·2.5 H2O S 7 – S 8

DEPT spectra of compound 1·1.5 Na·2.5 H2O S 9 – S 10

HMBC spectra of compound 1·1.5 Na·2.5 H2O S 11 – S 17

MASS

PATENT

WO 2004071503

WO 2005005372

WO 2006016657

WO 2006088071

WO 2007000994

WO 2007132882

WO 2009154248

WO 2014027696

PAPER

Angewandte Chemie, International Edition (2012), 51(17), 4218-4222, S4218/1-S4218/77.

Bioorganic & Medicinal Chemistry Letters (2013), 23(1), 336-339

PAPER

Organic & Biomolecular Chemistry (2017), 15(31), 6632-6639.

http://pubs.rsc.org/en/Content/ArticleLanding/2017/OB/C7OB01608E#!divAbstract

10.1039/C7OB01608E

Stereoselective synthesis of the viridiofungin analogue NA808 from a chiral tetrahydrofuran-carboxylic acid

Masatoshi Murakata*^a  and  Takuma Ikeda^a 

 Author affiliations

*Corresponding authors

^aAPI Process Development Department, Chugai Pharmaceutical Co., Ltd., 5-5-1 Ukima, Kita-Ku, Japan
E-mail: murakatamst@chugai-pharm.co.jp

Abstract

The viridiofungin analogue NA808 was synthesized by the stereoselective Ireland–Claisen rearrangement of dienylmethyl ester, regioselective bromolactonization of β-divinylpropanoic acid and retro-bromolactonization.

http://www.rsc.org/suppdata/c7/ob/c7ob01608e/c7ob01608e1.pdf

  

PATENT

https://patents.google.com/patent/WO2004071503A1/ar

The number of people infected with hepatitis C virus (HCV) is estimated at 1 to 200 million people worldwide, and over 2 million people in Japan. Approximately 50% of these patients migrate to chronic hepatitis, of which approximately 20% become liver cirrhosis, liver cancer after more than 30 years after infection. About 90% of liver cancer is said to be hepatitis C cause. In Japan, more than 20,000 patients die every year from liver cancer associated with HCV infection.

HCV was discovered in 1989 as a major causative virus of non-A non-B hepatitis after transfusion. HCV is an enveloped RNA virus whose genome

It consists of single-stranded (+) RNA and is classified as a genus Hepacivirus of Flaviviridae.

Since HCV avoids the immune mechanism of the host due to a cause which is still unclear, persistent infection is often established even when infected with an adult with developed immune mechanism, progresses to chronic hepatitis, liver cirrhosis, hepatocellular carcinoma, surgery It is also known that many patients have liver cancer recurrence due to inflammation that continues to occur in non-cancerous areas.

Therefore, establishment of an effective therapy for hepatitis C is desired, and among them, apart from coping therapy that suppresses inflammation by anti-inflammatory agents, development of a drug that reduces or eradicates HCV in the affected liver It is strongly desired.

Interferon treatment is currently known as the only effective treatment for HCV elimination. However, the number of patients with interferon effective is about one third of all patients. In particular, interferon response to HCV genotype 1 b is very low. Therefore, development of anti-HCV drugs that can replace or be used in combination with interferon is strongly desired.

In recent years, Ribavirin (1 – 3 – D – lipofuranosyl – 1 H – 1, 2, 4 – triazole – 3 – carboxamide) is commercially available as a therapeutic agent for hepatitis C by combining with interferon, Is still low, further new treatment for hepatitis C is desired. In addition, attempts have been made to eliminate viruses by enhancing the immune system of patients, such as interferon agonists, interleukin-12 agonists, etc. However, no effective drug has yet been found.

Since the HCV gene has been cloned, molecular biological analysis of the mechanism and function of viral genes, functions of proteins of each virus and the like has been accompanied by rapid development of forces, replication of virus in host cells, persistent infection, pathogenesis The mechanism such as sexuality has not been sufficiently elucidated, and at the present time, an HCV infection experiment system using reliable cultured cells has not been constructed. Conventionally, when evaluating anti-HCV drugs, alternative alternative virus method using other closely related viruses had to be used.

In recent years, however, it became possible to observe in vitro HCV replication using the nonstructural region part of HCV, so that anti-HCV drugs could be easily evaluated by the replicon assay method (Non-Patent Document 1). The mechanism of H CV RN A replication in this system is believed to be identical to the replication of the full-length HCV RNA genome infected with hepatocytes. Therefore, this system can be said to be a cell-based approach system useful for identifying compounds that inhibit the replication of HCV.

The compounds claimed in this patent are compounds that inhibit the replication of HCV found by the replicon astrocyte method. These inhibitors are considered highly likely to be therapeutic agents for HCV.

Non-Patent Document 1

B. Roman et al., Science (Science), 1999, 285, 110 – 113

Example 14

– 1 (Step 1 1)

According to the method described in the literature (J. Org. Chem. 1989, 45, 5522, BE Marron, et al)

TBDPSO.

a on

Of compound a (7.0 1 g) was synthesized, and anhydrous ethyl ether of this compound a

(700 ml) was cooled to 0 ° C. and bis (2-methoxyethoxy) aluminum hydride (414 mmol, 218 ml, 70% toluene solution) was added slowly. Five minutes after adding the reagent, the ice bath was removed and stirring was continued for 1 hour at room temperature. The reaction solution was cooled to 0 ° C and anhydrous ethyl acetate (1 9.8 ml, 203 mmol) was added slowly. After stirring at the same temperature for 10 minutes, it was cooled to 1 78 ° C., and iodine (76.1 g,

300 thigh ₀ 1) was added. The temperature was gradually raised to room temperature over 2 hours to complete the reaction. To the reaction solution was added aqueous sodium bisulfite solution, and ethyl acetate was added. The reaction solution was filtered with suction through celite, the organic layer was separated, and the aqueous layer was extracted again with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude title compound (100 g) as a light brown oily substance. The obtained crude product was directly used for the next reaction.

Physicochemical properties of compound b

Molecular weight 466

FAB-MS (positive mode, matrix m-NBA) 467 (M + H ⁺ ).

Chemical shift value of ^X H-NMR (in heavy chloroform) δ:

J = 6 Hz), 3.80 (2H, t, J = 6 Hz), 4.18 (2H, t, J = 5 Hz), 2.73 (2H, t, J = 6 Hz), 1.49 Hz, 5.91 (1 H, t, J = 5 Hz), 7.35 – 7.46 (6 H, m), 7.65 – 7.69 (4 H, m)

1 -2 (Step 1 – 2)

TBDPS

Dichloro port methane solution of compound b obtained in the above reaction (300 ml) was cooled to 0 ° C, dihydropyran (22. 7 ml, 248删₀ plus 1). Pyridinium paratoluenesulfonic acid (260 mg, 1 mol) was added to this solution. After 1 hour sodium bicarbonate water was added to stop the reaction. The separated organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound c (108 g) thus obtained was directly used for the next reaction.

Physicochemical properties of compound c

Molecular weight 550

FAB-MS (positive mode, matrix m-NBA) 551 (M + H ⁺ )

Chemical shift value of ¹ H – NMR (in heavy mouth formium) δ:

3.46-3.58 (2H, m), 3.76 (2H, t, J = 6 Hz), 3.82 (2H, t, J = 6 Hz), 1.04 (9H, s), 1.49-1.91 J = 13, 6 Hz), 4.65 (1 H, t, J = 3 Hz), 5.91 (1 H, t (s)), 4.93 (1 H, m), 4.06 (1 H, dd, J = 13, 6 Hz) , J = 5 Hz) 7.35 – 7.43 (6 H, m), 7.65 – 7.69 (4 H, m)

1-3 (Step 1- 3)

The crude compound c (4. 73 g) was dissolved in anhydrous ethyl ether (30 ml) and cooled to 1 78 ° C. Tert-butyllithium (1 7. 2 mol, 1 0.7 ml, 1.6 N pentane solution) was added slowly. After stirring at the same temperature for 1 hour, paraformaldehyde (1 8.9 mraol, 570 mg) was added and the mixture was warmed to 0 ° C. for 30 minutes at the same temperature and stirred for 1 hour. An aqueous solution of salthyanmonium was added to stop the reaction, and the mixture was extracted with ethyl acetate. The aqueous layer was extracted with a small amount of ethyl acetate and the combined organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The crude product obtained by concentration under reduced pressure was purified by column chromatography (silica gel, hexane-ethyl acetate 9: 1 to 4: 1) to give compound d (1. 635 g) as a colorless oily substance.

Physicochemical properties of compound d

Molecular weight 454

FAB-MS (positive mode, matrix m-NBA) 455 (M + H ⁺ )

^ – NMR (chemical shift value in heavy chloroform) δ:

J = 6 Hz), 3.03 (1 H, t, J = 6 Hz), 3.47 – 3.58 (2 H, m), 3.75 – 3.92 (2 H, (3 H, m), 4.08 – 4.26 (4 H, m), 4.68 (1 H, t, 3 Hz),

5.53 (1 H, t, J = 7 Hz) 7.35 – 7.47 (6 H, m), 7.64 – 7.68 (4 H, m)

1 -4 (Step 1 – 4)

An anhydrous N, N-dimethylformamide solution (2 ml) of the compound d (34 mg, 0. 76 mmol) and imidazole (71 mg, 1.14 mmol) was cooled to 0 ° C and tert- Chlorosilane (0: 2 ml, 0. 76 mmol) was capped and stirred for 2 hours. An ammonium chloride aqueous solution was added to stop the reaction, and the mixture was extracted with hexane. The organic layer was washed with water twice, followed by saturated brine and dried over anhydrous sodium sulfate. And concentrated under reduced pressure to obtain crude compound e (554 mg) as a colorless oily substance.

Physicochemical properties of compound e

FAB-MS (positive mode, matrix m-NBA) 715 (M + Na ⁺ )

Chemical shift value of ‘1 H-NMR (in heavy mouth formium) δ:

(4H, m), 1.00 J = 7 Hz), 5.43 (1 H, t, J = 7 Hz), 7.29 – 7.48 (12 H, m), 4.00 – 4.09 (1 H, m), 4.14 , 7.57 – 7.78 (8 H, m)

1-5 (Step 1- 5)

f

Pyridinium paratoluenesulfonic acid (9 O mg, 0.36 mmol) was added to an ethanol solution (6 ml) of the compound e (1. 16 g, 1. 67 mmol), and the mixture was stirred at 60 ° C. for 3.5 hours. After cooling the solution to room temperature, a saturated aqueous sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic layer was washed successively with water and saturated brine, and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and the resulting crude product was purified by column chromatography (silica gel, hexane / ethyl acetate 20: 1) to give compound f (825 mg, 81%) as a colorless oily substance.

Physicochemical properties of compound f

Molecular weight 608

FAB-MS (positive mode, matrix m-NBA) 631 (M + Na ⁺ )

^ – NMR (chemical shift value in heavy chloroform) δ:

(2H, t, J = 7 Hz), 3.75 (2H, t, J = 7 Hz), 3.90 (2H, t, J = 7 Hz), 1.01 (9H, s), 1.01 , 7.59-7.47 (12 H m), 7.57-7.75 (8 H, m), 4.14 (2 H, s), 5 47 (1 H, t, J =

1-6 (Step 1-6)

9

The round bottom flask containing the rotor was heated and dried under reduced pressure and then purged with nitrogen, and anhydrous

Dichloromethane (60 ml) was added and cooled to _20 ° C. Titanium tetraisopropoxide (2.3 3 ml, 7.8 8 mmol), L 1 (+) – Jetyl tartrate (1.6 2 ml, 9. 4 6 min. ₀ 1) was added successively, and after stirring for 15 minutes, compound f (4.80 g, 7. 88 mmol) in dichloromethane (30 ml), and the mixture was stirred for 15 minutes. Cool to _ 25 ° C and add tert-butyl hydroperoxide (5. 25 ml,

15. 8 mmol, 3 N dichloromethane solution) was slowly added dropwise. After completion of the dropwise addition, the mixture was stirred at 20 ° C. for 2 hours, dimethylsulfide (1.1 ml) was added, and the mixture was further stirred at the same temperature for 1 hour. A 10% aqueous solution of tartaric acid was added to the reaction solution and the mixture was stirred for 30 minutes, and then stirred at room temperature for 1 hour. The organic layer was separated, the aqueous layer was extracted with a small amount of dichloromethane and the combined organic layers were dried over anhydrous sodium sulfate. The crude product obtained was concentrated under reduced pressure, and purified by force RAM chromatography (silica gel, hexane / monoacetic acid ethyl 9: 1). Compound g (4. 78 g, 97%) was obtained as a colorless oily substance. The asymmetric yield (> 95% ee) was determined by NMR analysis of the corresponding MT PA ester.

Physicochemical properties of compound g

Molecular weight 624

F AB-MS (positive mode, matrix m-NBA) 647 (M + Na ⁺ )

– Chemical shift value of NMR (in heavy chloroform) δ:

J = 14, 7 Hz), 2.23 (1 H, dt, J = 14, 1 H), 1.02 (9 H, s), 1.03 (9 H, s), 1.72 (6H, m), 7.32-7.45 (12H, m), 7.60- 7.65 (8H, m), 6.5 Hz), 3.17 (1H, dd, J = 6, 5 Hz), 3.55-3.79

1 – 7 (Step 1 – 7)

To a solution (100 ml) of the compound α (10. 45 g, 37.2 mmol) produced in the step 2-3 of Production Example 1 described below in an anhydrous tetrahydrofuran solution (100 ml) under a nitrogen atmosphere was added biscyclopentadienylzirconium hydride chloride (10. lg, 37.2 mol) was added at room temperature and stirred for 30 minutes. The resulting solution was cooled to 1780C and methyl magnesium chloride (24.7 ml, 74 mmol, 3 N tetrahydrofuran

Furan solution), and the mixture was stirred for 5 minutes. Monovalent copper iodide (500 mg, 7.2 mM) was added to this solution and the temperature was gradually raised to _ 30 ° C. An anhydrous tetrahydrofuran solution (70 ml) of the compound g (4. 49 g) was added over 20 minutes, and after completion of the dropwise addition, the mixture was stirred at 25 ° C. overnight. The saturated ammonium chloride aqueous solution was slowly added, the reaction was stopped, and the temperature was gradually raised to room temperature. The mixture was stirred at room temperature for 10 hours and the resulting white solid was filtered off through celite. The celite was washed thoroughly with ethyl acetate and the organic layer was separated. The aqueous layer was extracted with a small amount of ethyl acetate and the combined organic layer was washed with saturated aqueous ammonium chloride solution and then dried over anhydrous sodium sulfate. Concentrated under reduced pressure and the obtained crude product was purified by column chromatography (silica gel, hexyl acetate

20: 1 to 9: 1) to give compound h (5. 96 g, 91%) as a pale yellow oily substance.

Physicochemical properties of compound h

Molecular weight 907

F AB – MS (negative mode, matrix πι – Α Β A) 906 (Μ – Η ⁺ )

Chemical shift value of ¹ H-NMR (in heavy chloroform) δ:

0.88 (3H, t, 
0.99 (9H, s), 1.04 (9H, s), 1.18-1.63 (22H, m), 1.78-2.01 (4H, m), 2.44-2.57 (1H, m), 3.00 (1H, t, J = 6 Hz), 3.59-3.92 (10H, m), 4.28 (1H, s), 5.37-5.55 (2H, m), 7.29-7.65 (20H, m)

1-8 (Step 1-8)

Compound h (5.30 g, 5.84 dragon ol) was dissolved in dichloromethane (200 ml) and 2, 2-dimethoxypropane (150 ml), pyridinium paratoluenesulfonic acid (15 mg, 0.058 mmol) was added , And the mixture was stirred at room temperature overnight. The reaction was quenched by adding saturated aqueous sodium bicarbonate and extracted twice with dichloromethane. After drying over anhydrous sodium sulfate, the mixture was concentrated under reduced pressure, and the resulting crude product was purified by column chromatography (silica gel, hexane-ethyl acetate 20: 1). Compound i (4. 69 g, 86%) was obtained as a pale yellow oily substance.

Physicochemical properties of Compound i

Molecular weight 947

F AB-MS (negative mode, matrix m-NBA) 946 (M – H ⁺ )

Chemical shift value of ¹ H – NMR (in heavy mouth formium) δ:

(1 H, m), 0.88 (3H, t, J = 6 Hz), 1.02 (9H, s), 1.05 (9H, s), 1.14-1.63 (28H, m), 1. 2.16 (2H, m), 7.28 – 7.47 (12H, m), 7.61 – 7.69 (1H, d, J = 10 Hz), 3.64-3.86 (6H, m 3.92 (s, 4H), 5.36-5.42 8 H, m) 1 – 9 (Step 1 – 9)

A tetrahydrofuran solution (50 ml) of the compound i (4. 39 g, 4. 64 mmol) was cooled to 0 ° C., tetrabutylammonium fluoride (10. 2 ml, 10, 2 difficulty, 1 M tetrahydrofuran solution) and Acetic acid (0. 53 ml, 9. 27 mmol) was added. The temperature was gradually raised to room temperature and stirred for 2 days. A saturated ammonium chloride aqueous solution was added and the mixture was extracted twice with dichloromethane. The combined organic layer was washed with aqueous sodium bicarbonate and dried over anhydrous sodium sulfate. The crude product was purified by column chromatography (silica gel, hexane-ethyl acetate 9: 1 to 3: 2) to obtain the compound〗 (1. 73 g, 81%) Was obtained as a pale yellow oily substance.

Physicochemical properties of compound j

Molecular weight 470

F AB-MS (positive mode, matrix m-NBA) 493 (M + Na ⁺ )

Chemical shift value of ^X H-NMR (in heavy chloroform) δ:

2.73 (1H, dt, J = 6, 10 Hz), 2.95 (3H, t, J = 6 Hz), 1.17-1.73 (26H, m), 1.91-2.16 (4H, m), 2.4 J = 15, 7 Hz (1 H, dt, J = 15 Hz), 3.48 (1 H, d, J = 1 Hz), 3.63-4.01 (m, 10 H), 5.15 )

1-10 (Step 1- 10)

Under an atmosphere of nitrogen, a solution of oxalyl chloride (0. 575 ml, 6. 6 mol) in anhydrous dichloromethane (17 ml) was cooled to 178 ° C. and dimethyl sulfoxide

(0. 9 36 ml, 1 3 2 minol) in dichloromethane (1 ml) was added dropwise and the mixture was stirred for 15 minutes. Dichloromethane solution (5 ml) of compound j (388 mg, 0. 824 aura) was slowly added dropwise. The mixture was stirred at the same temperature for 1 hour, then terethylamine (3 ml, 21.4fflmol) was added and the mixture was stirred for 30 minutes. The cooling bath was removed and a low-boiling compound was removed by blowing a nitrogen gas stream to the solution, followed by drying under reduced pressure. Jether ether (15 ml) was added to the residue, and insoluble matter was filtered off and concentrated. After this operation was carried out twice, the obtained residue was immediately used for the next reaction.

The crude dialdehyde was dissolved in 2-methyl-2-propanol (24 ml) and 2-methyl-2-butene (6 ml) and cooled to about 5 to 7 ° C. To this solution was added sodium chlorite (745 mg, 8. 24 mmol) and sodium dihydrogenphosphate

(745 mg, 6. 2 l mmol) in water (7. 45 ml) was slowly added dropwise. After 2 hours the mixture was cooled to 0 ° C. and aqueous sodium hydrogenphosphate solution was added to adjust PH to approximately 5. The mixture was extracted three times with dichloromethane, and the combined organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate. After filtration, concentration under reduced pressure afforded a pale yellow oily residue which was immediately used for the next reaction without further purification.

The crude dicarboxylic acid was dissolved in N, N-dimethylformamide di tert-butylacetal (4. 5 ml) and stirred at 70 ° C. for 1 hour. The low boiling point compound was distilled off under reduced pressure. The residue was purified by column chromatography (silica gel, hexane / ethyl acetate 20: 1) to give compound k (340 mg, 60%) as a pale yellow oily substance.

Physicochemical properties of compound k

Molecular weight 6 10

FAB-MS (positive mode, matrix m-NBA) (M + H ⁺ ) 611, (M + Na ⁺ ) 633

^ – NMR (chemical shift value in heavy chloroform) δ:

(2H, ABq, J = 15, 18 Hz), 2.93 (1 H, q, J = 6 Hz), 1.18 J = 7 Hz), 3.82-3.88 (2H, m), 3.92 (4H, s), 5.51-5.69 (2H, m)

1- 11 (Step 1 – 1 1)

Compound k (34 mg, 0. 556 mmol) was dissolved in tetrahydrofuran (1 ml), 80% acetic acid aqueous solution (10 ml) was added, and the mixture was stirred at room temperature for 3.5 hours. The mixture was slowly added into a saturated aqueous solution of sodium bicarbonate to neutralize acetic acid and then extracted twice with ethyl acetate. Drying over anhydrous sodium sulfate, followed by filtration and concentration under reduced pressure to give compound t

(290 mg, 99%) as a pale yellow oil.

Physicochemical properties of compound f

Molecular weight 526

FAB – MS (positive mode, matrix m – NBA) (M + H ⁺ ) 527,

(M + Na ⁺ ) 549

Chemical shift value of iH-NMR (in heavy chloroform) δ:

(2H, Q 
, 2.25-2.41 (5H, m), 1.99 (1H, d, J = 7 Hz), 2.04 (1H, d (1H, t, 7 Hz), 1.18- 1.68 (36H, ra), 2.01 J = 7 Hz), 5.58 (1 H, dt, J = 16, 6 Hz), 3.62 (3H, m), 3.99 (1H, s), 5.42

1-12 (Step 1 – 12)

Acetone (45 ml) was cooled to 0 ° C. and Jyones reagent (0.48 ml, 0.9 mmol, 1.8 9 N) was added. An acetone solution (3 ml) of the compound (216 mg, 0, 41) was slowly added dropwise to this mixture. Stirring at the same temperature for 1 hour

After stirring, the reaction was stopped by adding an aqueous sodium bisulfite solution until the yellow color of the reaction disappeared and a dark green precipitate appeared. A saturated saline solution (20 ml) was added thereto, and the mixture was extracted twice with dichloromethane, and the combined organic layer was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane monomethanol 50: 1 to 20: 1) to give compound m (198 mg, 89%) as a pale yellow oily substance.

Physicochemical properties of compound m

Molecular weight 541

ESI (L CZMS positive mode) (M + H ⁺ ) 542

Chemical shift value of ¹ H – NMR (in heavy mouth formium) δ:

J = 8 Hz), 2.70 (1 H, t, J = 6 Hz), 1.16 – 1.67 (36 H, m), 1.99 (2 H, J = 15, 5 Hz), 2.68 (1 H, d, J = 9 Hz), 3.28 )

1 – 13 (Step 1 – 13)

A solution of the compound m (6. 4 mg, 0.12 mmol), a solution of (S) -4- (2-butynyloxy) phenylalanine t-butyl ester hydrochloride (4.6 mg, 0.114 mmol) in N, N-dimethylformamide lml) was cooled to 110 ° C and N, N-diisopropylethylamine (0 ° 5 ml, 0.026 mmol), O- (7-azobenzotriazole 1- 1, N, N, N ‘, N’ – tetramethyluronium hexafluorophosphate (7.0 mg, 0.17 mmol) was added sequentially. The temperature was raised to room temperature with stirring and stirred overnight. An aqueous ammonium chloride solution was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic layer was washed twice with water and then with saturated brine, and then dried over anhydrous sodium sulfate. After filtration and concentration under reduced pressure, the residue was purified by thin layer silica gel thin layer chromatography (hexane / ethyl acetate 7: 3) to obtain compound n

(8. 4 mg, 88%) as a colorless solid.

Physicochemical properties of compound n

^ – NMR (chemical shift value in heavy chloroform) δ:

J = 1.9 Hz), 1.90-2.03 (2H, m), 2.29 – 2.43 (4H, t, J = 6.9 Hz), 1 .12-1.68 (45H, m), 1.85 (3H, m), 4.22 (1 H, s), 4.57 – 4.74 (3H, d, J = 16.5 Hz) J = 8.6 Hz), 7.01 (1 H, d, J = 8.6 Hz), 5.46 (1 H, dd J = 9.2, 15.2 Hz), 5.64 (1 H, dt, J = 6.6, 15.2 Hz) 7.9 Hz), 7.13 (2H, d, J – 8.6 Hz)

1-14 (Step 1 – 14)

Dichloromethane solution (3 ml) of compound n (8.4 mg) was cooled to 0 ° C and anisanol (0.01 ml) and trifluoroacetic acid (1 ml) were sequentially added. Slowly warmed to room temperature and stirred overnight. After concentrating the reaction solution under reduced pressure and azeotropically twice with benzene, the residue was purified with megabond-1-butanediol (500 mg, Parian) (dichloromethane-methanol = 20: 1) to obtain Compound 21 (5. 3 mg, 80% As a colorless solid.

Physicochemical properties of compound 21 ‘

Molecular weight 643

ESI (LC / MS positive mode) 644 (M + H +)

Chemical shift value of ¹ H – NMR (in methanol d – 4) δ:

0.90 (3 H, t, J = 7 Hz), 1.19 – 1.38 (1 m), 1.42 – 1.60 (cm), 1.82 (3 H, t,

J = 2 Hz), 2.8 – 2.98 (2 H, m), 3.09 – 3.23 (2 H, m), 2.8 (2H, d, J = 9 Hz) 7 7.13 (2H, d, J = 9 Hz), 4.53 – 4.67 (3H, m), 5.39-5.61 (2H, m), 6.83

Patent ID	Patent Title	Submitted Date	Granted Date
US2011098477	Method Of Producing Compound Having Anti-Hcv Activity	2011-04-28
US2010152457	Intermediate compound for synthesis of viridiofungin a derivative	2010-06-17
US8030496	Intermediate compound for synthesis of viridiofungin a derivative	2010-06-17	2011-10-04
US7897783	Intermediate compound for synthesis of viridiofungin a derivative	2008-11-27	2011-03-01

 

Patent ID	Patent Title	Submitted Date	Granted Date
US2011160252	PHARMACEUTICAL COMPOSITIONS FOR TREATMENT OR PREVENTION OF HBV INFECTION	2011-06-30
US2010274026	Virus therapeutic drug	2010-10-28
US7776918	Remedy for viral disease	2006-09-28	2010-08-17
US7378446	Compound having anti-hcv activity and process for producing the same	2006-08-31	2008-05-27
US9266853	ORALLY AVAILABLE VIRIDIOFUNGIN DERIVATIVE POSSESSING ANTI-HCV ACTIVITY	2013-08-16	2015-07-30

 

References

Discovery of NA808: A novel host targeting anti-HCV agent
237th Am Chem Soc (ACS) Natl Meet (March 22-26, Salt Lake City) 2009, Abst MEDI 14

///////////////CH4630808, CH 4630808, NA 808

Viridiofungin A

CCCCCCCC(=O)CCCCCCC=CC(C(=O)NC(CC1=CC=C(C=C1)O)C(=O)O)C(CC(=O)O)(C(=O)O)O

TITLE COMPD

O=C(O)[C@](O)(CC(=O)O)[C@H](\C=C\CCCCCCC(=O)CCCCCCC)C(=O)N[C@@H](Cc1ccc(OCC#CC)cc1)C(=O)O

Doconexent

CAS 6217-54-5

WeightAverage: 328.4883
Chemical FormulaC₂₂H₃₂O₂

4,7,10,13,16,19-Docosahexaenoic acid, (4Z,7Z,10Z,13Z,16Z,19Z)-

Doconexent sodium

295P7EPT4C

81926-93-4

• (4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic acid
• 22:6-4, 7,10,13,16,19
• 22:6(n-3)
• 4,7,10,13,16,19-docosahexaenoic acid
• 4,7,10,13,16,19-Docosahexaenoic acid
• all-cis-4,7,10,13,16,19-docosahexaenoic acid
• all-cis-DHA
• cervonic acid
• DHA
• docosa-4,7,10,13,16,19-hexaenoic acid
• Docosahexaenoic acid
• Ropufa 60
• S.Presso
• all-Z-Docosahexaenoic acid
• all-cis-4,7,10,13,16,19-Docosahexaenoic acid
• Δ^{4,7,10,13,16,19}-Docosahexaenoic acid
• 4,7,10,13,16,19-Docosahexaenoic acid, (all-Z)- (8CI)
• Docosahexaenoic acid (6CI)
• • (4Z,7Z,10Z,13Z,16Z,19Z)-4,7,10,13,16,19-Docosahexaenoic acid
  • (4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic acid
  • (4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexenoic acid
  • (all-Z)-4,7,10,13,16,19-Docosahexaenoic acid
  • 4-cis,7-cis,10-cis,13-cis,16-cis,19-cis-Docosahexaenoic acid

A mixture of fish oil and primrose oil, doconexent is used as a high-docosahexaenoic acid (DHA) supplement. DHA is a 22 carbon chain with 6 cis double bonds with anti-inflammatory effects. It can be biosythesized from alpha-linolenic acid or commercially manufactured from microalgae. It is an omega-3 fatty acid and primary structural component of the human brain, cerebral cortex, skin, and retina thus plays an important role in their development and function. The amino-phospholipid DHA is found at a high concentration across several brain subcellular fractions, including nerve terminals, microsomes, synaptic vesicles, and synaptosomal plasma membranes

Synthesis , By Farmer, Ernest H.; Van den Heuvel, Frantz A., From Journal of the Chemical Society (1938), 427-30.

ALSO

Docosahexaenoic acid (DHA) is an omega-3 fatty acid that is a primary structural component of the human brain, cerebral cortex, skin, and retina. It can be synthesized from alpha-linolenic acid or obtained directly from maternal milk (breast milk), fish oil, or algae oil.^[1]

DHA’s structure is a carboxylic acid (-oic acid) with a 22-carbon chain (docosa- derives from the Ancient Greek for 22) and six (hexa-) cis double bonds (-en-);^[2] with the first double bond located at the third carbon from the omega end.^[3] Its trivial name is cervonic acid, its systematic name is all-cis-docosa-4,7,10,13,16,19-hexa-enoic acid, and its shorthand name is 22:6(n−3) in the nomenclature of fatty acids.

Most of the DHA in fish and multi-cellular organisms with access to cold-water oceanic foods originates from photosynthetic and heterotrophic microalgae, and becomes increasingly concentrated in organisms the further they are up the food chain. DHA is also commercially manufactured from microalgae: Crypthecodinium cohnii and another of the genus Schizochytrium.^[4] DHA manufactured using microalgae is vegetarian.^[5]

In strict herbivores, DHA is manufactured internally from α-linolenic acid, a shorter omega-3 fatty acid manufactured by plants (and also occurring in animal products as obtained from plants), while omnivores and carnivores primarily obtain DHA from their diet.^[6] Limited amounts of eicosapentaenoic and docosapentaenoic acids are possible products of α-linolenic acid metabolism in young women^[7] and men.^[6] DHA in breast milk is important for the developing infant.^[8] Rates of DHA production in women are 15% higher than in men.^[9]

DHA is a major fatty acid in brain phospholipids and the retina. While the potential roles of DHA in the mechanisms of Alzheimer’s disease are under active research,^[10] studies of fish oil supplements, which contain DHA, have failed to support claims of preventing cardiovascular diseases.^[11][12][13]

Central nervous system constituent

DHA is the most abundant omega-3 fatty acid in the brain and retina. DHA comprises 40% of the polyunsaturated fatty acids (PUFAs) in the brain and 60% of the PUFAs in the retina. Fifty percent of the weight of a neuron‘s plasma membraneis composed of DHA.^[14]

DHA modulates the carrier-mediated transport of choline, glycine, and taurine, the function of delayed rectifier potassium channels, and the response of rhodopsin contained in the synaptic vesicles, among many other functions.^[15]

DHA deficiency is associated with cognitive decline.^[16] Phosphatidylserine (PS) controls apoptosis, and low DHA levels lower neural cell PS and increase neural cell death.^[17] DHA levels are reduced in the brain tissue of severely depressed patients.^[18][19]

Metabolic synthesis

In humans, DHA is either obtained from the diet or may be converted in small amounts from eicosapentaenoic acid (EPA, 20:5, ω-3) via docosapentaenoic acid (DPA, 22:5 ω-3) as an intermediate.^[7][6] This synthesis had been thought to occur through an elongation step followed by the action of Δ4-desaturase.^[6] It is now considered more likely that DHA is biosynthesized via a C24 intermediate followed by beta oxidation in peroxisomes. Thus, EPA is twice elongated, yielding 24:5 ω-3, then desaturated to 24:6 ω-3, then shortened to DHA (22:6 ω-3) via beta oxidation. This pathway is known as Sprecher’s shunt.^[20][21]

In organisms such as microalgae, mosses and fungi, biosynthesis of DHA usually occurs as a series of desaturation and elongation reactions, catalyzed by the sequential action of desaturase and elongase enzymes. A common pathway in these organisms involves:

1. a desaturation at the sixth carbon of alpha-linolenic acid by a Δ6 desaturase to produce stearidonic acid,
2. elongation of the stearidonic acid by a Δ6 elongase to produce to eicosatetraenoic acid,
3. desaturation at the fifth carbon of eicosatetraenoic acid by a Δ5 desaturase to produce eicosapentaenoic acid,
4. elongation of eicosapentaenoic acid by a Δ5 elongase to produce docosapentaenoic acid, and
5. desaturation at the fourth carbon of docosapentaenoic acid by a Δ4 desaturase to produce DHA.^[22]

Metabolism

DHA can be metabolized into DHA-derived specialized pro-resolving mediators (SPMs), DHA epoxides, electrophilic oxo-derivatives (EFOX) of DHA, neuroprostanes, ethanolamines, acylglycerols, docosahexaenoyl amides of amino acids or neurotransmitters, and branched DHA esters of hydroxy fatty acids, among others.^[23]

The enzyme CYP2C9 metabolizes DHA to epoxydocosapentaenoic acids (EDPs; primarily 19,20-epoxy-eicosapentaenoic acid isomers [i.e. 10,11-EDPs]).^[24]

Potential health effects

Neurological research

While one human trial of 402 subjects lasting 18 months concluded that DHA did not slow decline of mental function in elderly people with mild to moderate Alzheimer’s disease,^[25] a similar trial of 485 subjects lasting 6 months concluded that algal DHA of 900 mg per day taken decreased heart rate and improved memory and learning in healthy, older adults with mild memory complaints.^[26]

In another early-stage study, higher DHA levels in middle-aged adults was related to better performance on tests of nonverbal reasoning and mental flexibility, working memory, and vocabulary.^[27]

One study found that the use of DHA-rich fish oil capsules did not reduce postpartum depression in mothers or improve cognitive and language development in their offspring during early childhood.^[28] Another systematic review found that DHA had no significant benefits in improving visual field in individuals with retinitis pigmentosa.^[29] A 2017 pilot study found that fish oil supplementation reduced the depression symptoms emphasizing the importance of the target DHA levels.^[30]

Pregnancy and lactation

It has been recommended to eat foods which are high in omega-3 fatty acids for women who want to become pregnant or when nursing.^[31] A working group from the International Society for the Study of Fatty Acids and Lipids recommended 300 mg/day of DHA for pregnant and lactating women, whereas the average consumption was between 45 mg and 115 mg per day of the women in the study, similar to a Canadian study.^[32] Despite these recommendations, recent evidence from a trial of pregnant women randomized to receive supplementation with 800 mg/day of DHA versus placebo, showed that the supplement had no impact on the cognitive abilities of their children at up to seven years follow-up.^[33]

Other research

In one preliminary study, men who took DHA supplements for 6–12 weeks had lower blood markers of inflammation.^[34]

Nutrition

Ordinary types of cooked salmon contain 500–1500 mg DHA and 300–1000 mg EPA per 100 grams.^[35] Additional rich seafood sources of DHA include caviar (3400 mg per 100 grams), anchovies (1292 mg per 100 grams), mackerel (1195 mg per 100 grams), and cooked herring(1105 mg per 100 grams).^[35] Brains from mammals are also a good direct source, with beef brain, for example, containing approximately 855 mg of DHA per 100 grams in a serving.^[36]

Discovery of algae-based DHA

In the early 1980s, NASA sponsored scientific research on a plant-based food source that could generate oxygen and nutrition on long-duration space flights. Certain species of marine algae produced rich nutrients, leading to the development of an algae-based, vegetable-like oil that contains two polyunsaturated fatty acids, DHA and arachidonic acid,^[37] present in some health supplements.

Use as a food additive

DHA is widely used as a food supplement. It was first used primarily in infant formulas.^[38] In 2004, the US Food and Drug Administration endorsed qualified health claims for DHA.^[39]

Some manufactured DHA is a vegetarian product extracted from algae, and it competes on the market with fish oil that contains DHA and other omega-3s such as EPA. Both fish oil and DHA are odorless and tasteless after processing as a food additive.^[40]

Studies of vegetarians and vegans

Vegetarian diets typically contain limited amounts of DHA, and vegan diets typically contain no DHA.^[41] In preliminary research, algae-based supplements increased DHA levels.^[42]While there is little evidence of adverse health or cognitive effects due to DHA deficiency in adult vegetarians or vegans, breast milk levels remain a concern for supplying adequate DHA to the developing fetus.^[41]

DHA and EPA in fish oils

Fish oil is widely sold in capsules containing a mixture of omega-3 fatty acids, including EPA and DHA. Oxidized fish oil in supplement capsules may contain lower levels of EPA and DHA.^[43][44]

Hypothesized role in human evolution

An abundance of DHA in seafood has been suggested as being helpful in the development of a large brain,^[45] though other researchers claim a terrestrial diet could also have provided the necessary DHA.^[46]

Patent

CN 106190872

https://patents.google.com/patent/CN106190872A/zh

PATENT

WO 2017038860

https://patents.google.com/patent/WO2017038860A1/en

[Example 1]
The raw EPA ethyl ester 1 of Comparative Example 1 containing EPA 96.7%, except for changing the temperature of the alkaline hydrolysis in 6 ° C., in the same manner as in Comparative Example 1 was alkaline hydrolysis.
That is, the starting EPA ethyl ester 1 2.50 g, ethanol 6.25 mL (4.92 g, 14.11 equivalents relative fatty acid), water 1.00 mL, 48 wt% sodium hydroxide aqueous solution 0.76 g ( 1.20 equivalents of base) was added a sample solution 3 was prepared against fatty acids. In sample liquid 3, moisture 1.40 g, i.e., was 10.27 equivalents relative fatty acid. The sample liquid 3, stirred for 24 hours 6 ° C., was subjected to hydrolysis treatment. Confirmed the completion of the reaction of the hydrolysis treatment, returned to the sample liquid 3 after treatment at room temperature, after transferred to a separatory funnel, and hexane was added 3.13 mL, purified water 2.50mL the sample liquid 3. When further adding 2.25g of hydrochloric acid, the sample solution 3 was separated into two layers of hexane and aqueous layers. The pH of the aqueous layer was 1.0.

The sample liquid 3 was stirred, then the mixture was allowed to stand, after removing the aqueous layer from the sample liquid 3, was further stirred with purified water 3.75mL the sample liquid 3 after removal. Hydrochloric acid was added small amount to adjust the pH of the aqueous layer to 1.0. Thereafter, the aluminum plate was washed with the same amount of purified water as rinsing liquid. Rinsing liquid is recovered after washing with water was repeatedly washed with water until neutral pH 6.0 ~ 7.0. The hexane layer was recovered from the sample liquid 3 after washing with water, the recovered hexane layer, the hexane was removed with an evaporator and vacuum, the EPA3 a composition containing free EPA was obtained 2.14 g.
Against EPA3, it was evaluated in the same manner as EPA1. The results are shown in Table 1 and Table 4.
The recovery was 93.8%. The resulting Gardner color of EPA3 is 2-, AnV 1.3, ethyl ester (EE) content 2790Ppm, conjugated diene acid content was 0.47%. Conjugated unsaturated fatty acids other than the conjugated diene acid was not detected. These physical property values are shown in Table 1. Note that the conjugated unsaturated fatty acids, only the conjugated diene acid shown in Table 1.

PATENT

WO-2018120574

Process for production of docosahexaenoic acid (DHA), by microbial fermentation of Schizochytrium limacinum . Discloses use of DHA for treating cardiovascular diseases, infertility or neurological diseases. See CN106635405 , claiming method for separating DHA from powder DHA grease by supercritical extraction method. Kingdomway lists that it produces DHA by microorganism fermentation.

The current commercial sources of DHA are mainly fish oil and microalgae. DHA extracted from traditional deep-sea fish oil is unstable due to the variety, season and geographical location of fish, and the content of cholesterol and other unsaturated fatty acids is high. The difference in length and degree of unsaturation of fatty acid chains is large, resulting in limited production and content of DHA. It is not high, it is difficult to separate and purify, and the cost is high. With the growing shortage of fish oil raw materials, it is difficult to achieve the widespread use of DHA, a high value-added product in the food and pharmaceutical industries. The production of DHA by microbial fermentation can overcome the defects of traditional fish oil extraction, can be used for mass production of DHA, continuously meet people’s needs, has broad application prospects, and has attracted the attention of scholars at home and abroad. The microbial fermentation method uses fermented microorganisms such as fungi and microalgae to produce DHA-containing algal oil, and refined to obtain essential oil with high DHA content. DHA-producing strains approved by the Ministry of Health include Schizochytrium sp., Ulkenia amoeboida, and Crypthecodinium cohnii.

The market share of DHA produced by microbial fermentation is increasing rapidly year by year. There is a trend to replace DHA of fish oil, improve the production technology and quality of microalgae DHA, and the prospect of entering the microalgae DHA market is broad.

The publication No. CN103882072A discloses a method for producing docosahexaenoic acid by using Schizochytrium, and the highest yield disclosed is a cell dry weight of 61.2 g/L, a DHA content of 55.07%, and a DHA yield of 22.17 g. /L. The publication No. CN101812484A discloses a method for fermenting DHA by high-density culture of Schizochytrium, which discloses a dry cell weight of 120-150 g/L and a DHA yield of 26-30 g/L, which is also reported. The highest production level of DHA produced by Schizochytrium sp. Although the DHA productivity has been greatly improved compared with the previous research, the industrial production of docosahexaenoic acid by using microalgae greatly reduces the production cost, increases the unit yield, and enables the method of microbial fermentation to produce DHA. Promotion and popularization are still far from enough.

There are three main methods for extracting DHA from the fermentation liquid of Schizochytrium, one is centrifugation, the other is organic solvent extraction, and the third is supercritical extraction. Centrifugation, such as the publication No. CN101817738B, discloses a method for extracting DHA from algae and fungal cells by separating the microalgae or fungal fermentation broth after fermentation by a separation system, and adjusting the pH of the sludge with an acid. 2.0-4.0, then control the temperature of the slime at 10 °C-20 °C, add anti-oxidant in the slime, and then carry out high-pressure homogenization and breaking through the high-pressure homogenizer; add the broken mud to the water, stir and feed The liquid was separated by a three-phase separator to obtain DHA grease. The invention adopts physical wall breaking and physical extraction methods, has simple process, high cell breakage, low temperature treatment of bacteria sludge and antioxidant treatment, can effectively protect the biological activity of algae and fungal cells, and the product is green and non-toxic. Residue. However, the quality of the oil layer after centrifugation of the invention is poor. In addition to the oil, it also contains impurities such as water, medium components and cell debris, which is not conducive to subsequent refining. In addition, the wastewater layer after centrifugation contains a large amount of slag and has a high COD. Difficult to handle or process is extremely costly. The organic solvent extraction method, such as the publication No. CN101824363B, discloses a method for extracting docosahexaenoic acid oil: the fermentation liquid containing docosahexaenoic acid is subjected to enzymatic breaking, and then an organic solvent is used first. The first stage water is divided, the cells are enriched, and the organic solvent is used for secondary extraction to obtain a crude oil. The method is simple in operation and low in equipment investment, but the method uses organic solvent for extraction, and the final product may have solvent residue, and the extraction process has safety hazards such as flammability and explosion. The supercritical extraction method, as disclosed in the publication No. CN102181320B, discloses a method for extracting bio-fermented DHA algae oil, comprising the following steps: a) drying the solid matter obtained by solid-liquid separation of the microalgae fermentation liquid to obtain a dried bacterial cell; b) extracting the dried cells with supercritical carbon dioxide as an extractant to obtain a carbon dioxide fluid; c) separating the carbon dioxide fluid under reduced pressure to obtain DHA algae oil. Experiments show that the DHA content of DHA algae oil obtained by the method provided by the invention is more than 40%, the extraction yield is only 85.23%, and the need to add ethanol as the extracting agent has certain safety risks and supercritical. The equipment is expensive and the extraction yield is not high.

In the prior art, the refining of DHA hair oil is mostly carried out by chemical refining technology, and the DHA hair oil is degummed, alkali refining, decolorized and deodorized to obtain DHA essential oil. Inevitably, there are some problems in the process technology. For example, in order to achieve the requirement of controlling low acid value, alkali refining usually adds excessive alkali, and some triglycerides are inevitably saponified; high COD wastewater produced by alkali refining will pollute the environment; Alkali refining requires high temperature treatment for a long time, which is easy to cause the product’s peroxide value and anisidine value to increase; the deodorization temperature is high, and the long time is easy to produce trans fatty acids.

Currently, there is still a need to develop new DHA production processes.

Fermentation culture

In the following Examples 1-13, unless otherwise specified, the seed medium formulations used were: glucose 3%, peptone 1%, yeast powder 0.5%, sea crystal 2%, and pH natural (the rest being water). The fermentation medium formula is: glucose 12%, peptone 1%, yeast powder 0.5%, sea crystal 2% (the rest is water).

Example 1

The Schizochytrium sp. ATCC 20888, Schizochytrium limacinum Honda et Yokochi ATCCMYA-1381, and Schizochytrium sp. CGMCC No. 6843 slope-preserved strains were respectively inserted into 400 mL of medium. The 2L shake flask was cultured at a temperature of 25 ° C at a rotation speed of 200 rpm for 24 hours to complete the activated culture of the strain. According to the inoculation amount of 0.4%, the shake flask seed solution was connected to the first-stage seed tank containing the sterilized medium, and the culture temperature was 28 ° C, the aeration amount was 1 vvm, the tank pressure was 0.02 MPa, and the stirring speed was 50 rpm for 30 hours to complete the first stage. Seeds are expanded and cultured. The seed liquid of the primary seed tank was connected to the secondary seed tank containing the sterilized medium according to the inoculation amount of 3%, and the culture temperature was 28 ° C, the aeration amount was 1 vvm, the tank pressure was 0.02 MPa, and the stirring speed was 75 rpm for 24 hours. Complete secondary seed expansion culture. The seed solution of the secondary seed tank was connected to a fermentor containing the sterilized medium according to a 3% inoculum.

The fermentation process has a culture temperature of 28 ° C, aeration of 1 vvm, a can pressure of 0.02 MPa, a stirring speed of 75 rpm, a carbon source containing 30% of the pretreated crude glycerin, a glucose concentration of 5 g/L, and a nitrogen source. Fermentation culture. During the fermentation process, the glucose concentration, pH, bacterial biomass, crude oil production and DHA yield of the fermentation broth were measured.

After 96 hours of culture, the fermentation was terminated. Table 1 below shows the biomass, crude oil production, DHA production and DHA productivity of the three strains cultured in the original culture mode. Table 2 below shows the mixed fat and fatty acid composition of the gas obtained after fermentation. Analysis results. The biomass, crude oil production and DHA production of CGMCC No.6843 are also shown in Figure 3.

Table 1: Fermentation results of different strains in the original culture mode

Table 2: 100m ³ fermenter original culture method

It can be seen from Table 1 and Table 2 that the yield and fatty acid composition of the three strains are different in the original culture mode, and the Schizochytrium sp. CGMCC No. 6843 is superior to the other two strains. Schizochytrid sp. (Schizochytrium sp. CGMCC No. 6843) was used as the starting strain to optimize the different culture methods.

PATENT

CN106635405

https://patents.google.com/patent/CN106635405A/zh

PATENT

WO2012153345

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2012153345

PAPER

NMR

Organic Chemistry 2014 vol. 2014  21 pg. 4548 – 4561

Patent

WO 2015162265

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2015162265&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

Patent

References

External links

• DHA / EPA Omega-3 Institute – Recent studies, overviews, and objective science.
• Docosahexaenoic acid (DHA) – University of Maryland Medical Center (UMMC)
• Docosahexaenoic acid – DHA ChemSub Online

REFERENCE

1. Calder PC: Omega-3 fatty acids and inflammatory processes. Nutrients. 2010 Mar;2(3):355-74. doi: 10.3390/nu2030355. Epub 2010 Mar 18. [PubMed:22254027]
2. Kim HY: Novel metabolism of docosahexaenoic acid in neural cells. J Biol Chem. 2007 Jun 29;282(26):18661-5. Epub 2007 May 8. [PubMed:17488715]
3. Picq M, Chen P, Perez M, Michaud M, Vericel E, Guichardant M, Lagarde M: DHA metabolism: targeting the brain and lipoxygenation. Mol Neurobiol. 2010 Aug;42(1):48-51. doi: 10.1007/s12035-010-8131-7. Epub 2010 Apr 28. [PubMed:20422316]
4. Butovich IA, Lukyanova SM, Bachmann C: Dihydroxydocosahexaenoic acids of the neuroprotectin D family: synthesis, structure, and inhibition of human 5-lipoxygenase. J Lipid Res. 2006 Nov;47(11):2462-74. Epub 2006 Aug 9. [PubMed:16899822]
5. Serhan CN, Gotlinger K, Hong S, Lu Y, Siegelman J, Baer T, Yang R, Colgan SP, Petasis NA: Anti-inflammatory actions of neuroprotectin D1/protectin D1 and its natural stereoisomers: assignments of dihydroxy-containing docosatrienes. J Immunol. 2006 Feb 1;176(3):1848-59. [PubMed:16424216]
6. Mas E, Croft KD, Zahra P, Barden A, Mori TA: Resolvins D1, D2, and other mediators of self-limited resolution of inflammation in human blood following n-3 fatty acid supplementation. Clin Chem. 2012 Oct;58(10):1476-84. Epub 2012 Aug 21. [PubMed:22912397]
7. Chen CT, Kitson AP, Hopperton KE, Domenichiello AF, Trepanier MO, Lin LE, Ermini L, Post M, Thies F, Bazinet RP: Plasma non-esterified docosahexaenoic acid is the major pool supplying the brain. Sci Rep. 2015 Oct 29;5:15791. doi: 10.1038/srep15791. [PubMed:26511533]
8. Pawlosky RJ, Hibbeln JR, Novotny JA, Salem N Jr: Physiological compartmental analysis of alpha-linolenic acid metabolism in adult humans. J Lipid Res. 2001 Aug;42(8):1257-65. [PubMed:11483627]
9. Pawlosky RJ, Hibbeln JR, Salem N Jr: Compartmental analyses of plasma n-3 essential fatty acids among male and female smokers and nonsmokers. J Lipid Res. 2007 Apr;48(4):935-43. Epub 2007 Jan 17. [PubMed:17234605]
10. Cederholm T, Salem N Jr, Palmblad J: omega-3 fatty acids in the prevention of cognitive decline in humans. Adv Nutr. 2013 Nov 6;4(6):672-6. doi: 10.3945/an.113.004556. eCollection 2013 Nov. [PubMed:24228198]
11. Guesnet P, Alessandri JM: Docosahexaenoic acid (DHA) and the developing central nervous system (CNS) – Implications for dietary recommendations. Biochimie. 2011 Jan;93(1):7-12. doi: 10.1016/j.biochi.2010.05.005. Epub 2010 May 15. [PubMed:20478353]
12. Kelley DS, Siegel D, Fedor DM, Adkins Y, Mackey BE: DHA supplementation decreases serum C-reactive protein and other markers of inflammation in hypertriglyceridemic men. J Nutr. 2009 Mar;139(3):495-501. doi: 10.3945/jn.108.100354. Epub 2009 Jan 21. [PubMed:19158225]
13. Arterburn LM, Hall EB, Oken H: Distribution, interconversion, and dose response of n-3 fatty acids in humans. Am J Clin Nutr. 2006 Jun;83(6 Suppl):1467S-1476S. [PubMed:16841856]

Docosahexaenoic acid

Names
IUPAC name (4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid
Other names cervonic acid DHA doconexent (INN)
Identifiers
CAS Number	6217-54-5
3D model (JSmol)	Interactive image
ChEBI	CHEBI:28125
ChEMBL	ChEMBL367149
ChemSpider	393183
ECHA InfoCard	100.118.398
IUPHAR/BPS	1051
PubChem CID	445580
UNII	ZAD9OKH9JC
InChI[show]
SMILES[show]
Properties
Chemical formula	C22H32O2
Molar mass	328.488 g/mol
Density	0.943 g/cm3
Melting point	−44 °C (−47 °F; 229 K)
Boiling point	446.7 °C (836.1 °F; 719.8 K)
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

///////////Docosahexaenoic acid (22:6(n-3)), ZAD9OKH9JC, доконексен, دوكونيكسانت , 二十二碳六烯酸 , Doconexent, 6217-54-5, cervonic acid, DHA, doconexent, 81926-93-4

• all-Z-Docosahexaenoic acid
• AquaGrow Advantage
• CCRIS 7670
• Cervonic acid
• DHA
• Doconexent
• Doconexento
• Doconexento [INN-Spanish]
• Doconexentum
• Doconexentum [INN-Latin]
• Docosahexaenoic acid (all-Z)
• Doxonexent
• Efalex
• Marinol D 50TG
• Martek DHA HM
• Monolife 50
• Ropufa 60
• UNII-ZAD9OKH9JC

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

6	Promega 308064-99-5 MW: 644.9746
7	4,7,10,13,16,19-Docosahexaenoic acid, (4E,7E,10E,13E,16E,19E)- 391921-09-8 MW: 328.4928
8	Algal DHA MW: 328.4928
9	Omega-3 Fatty Acids MW: 909.3808

4,7,10,13,16,19-Docosahexaenoic acid 2091-24-9 MW: 328.493
2	Doconexent [INN] 6217-54-5 MW: 328.4928
3	Docosahexaenoic acid, (Z,Z,Z,Z,Z,Z)- 32839-18-2 MW: 328.493
4	Doconexent sodium 81926-93-4 MW: 350.4749
5	(14C)Docosahexaenoic acid 93470-46-3 MW: 328.493

Acamprosate calcium

3-acetamidopropane-1-sulfonic acid

Acamprosate, sold under the brand name Campral, is a medication used along with counselling to treat alcohol dependence.^[1][2]

Acamprosate, also known by the brand name Campral, is a drug used for treating alcohol dependence. Acamprosate is thought to stabilize the chemical balance in the brain that would otherwise be disrupted by alcoholism, possibly by blocking glutaminergic N-methyl-D-aspartate receptors, while gamma-aminobutyric acid type A receptors are activated. Reports indicate that acamprosate only works with a combination of attending support groups and abstinence from alcohol. Certain serious side effects include allergic reactions, irregular heartbeats, and low or high blood pressure, while less serious side effects include headaches, insomnia, and impotence. Acamprosate should not be taken by people with kidney problems or allergies to the drug.

Acamprosate is thought to stabilize chemical signaling in the brain that would otherwise be disrupted by alcohol withdrawal.^[3] When used alone, acamprosate is not an effective therapy for alcoholism in most individuals;^[4] however, studies have found that acamprosate works best when used in combination with psychosocial support since it facilitates a reduction in alcohol consumption as well as full abstinence.^[2][5][6]

Serious side effects include allergic reactions, abnormal heart rhythms, and low or high blood pressure, while less serious side effects include headaches, insomnia, and impotence.^[7] Diarrhea is the most common side-effect.^[8] Acamprosate should not be taken by people with kidney problems or allergies to the drug.^[9]

Until it became a generic in the United States, Campral was manufactured and marketed in the United States by Forest Laboratories, while Merck KGaA markets it outside the US.

Medical uses

Acamprosate is useful when used along with counselling in the treatment of alcohol dependence.^[2] Over three to twelve months it increases the number of people who do not drink at all and the number of days without alcohol.^[2] It appears to work as well as naltrexone.^[2]

Contraindications

Acamprosate is primarily removed by the kidneys and should not be given to people with severely impaired kidneys (creatinine clearance less than 30 mL/min). A dose reduction is suggested in those with moderately impaired kidneys (creatinine clearancebetween 30 mL/min and 50 mL/min).^[1][10] It is also contraindicated in those who have a strong allergic reaction to acamprosate calcium or any of its components.^[10]

Adverse effects

The US label carries warnings about increased of suicidal behavior, major depressive disorder, and kidney failure.^[1]

Adverse effects that caused people to stop taking the drug in clinical trials included diarrhea, nausea, depression, and anxiety.^[1]

Other frequent adverse effects include headache, stomach pain, back pain, muscle pain, joint pain, chest pain, infections, flu-like symptoms, chills, heart palpitations, high blood pressure, fainting, vomiting, upset stomach, constipation, increased appetite, weight gain, edema, sleepiness, decreased sex drive, impotence, forgetfulness, abnormal thinking, abnormal vision, distorted sense of taste, tremors, runny nose, coughing, difficulty breathing, sore throat, bronchitis, and rashes.^[1]

Pharmacology

Pharmacodynamics

The pharmacodynamics of acamprosate is complex and not fully understood;^[11][12][13] however, it is believed to act as an NMDA receptor antagonist and positive allosteric modulator of GABA_A receptors.^[12][13]

Ethanol and benzodiazepines act on the central nervous system by binding to the GABA_A receptor, increasing the effects of the inhibitory neurotransmitter GABA (i.e., they act as positive allosteric modulators at these receptors).^[12][4] In chronic alcohol abuse, one of the main mechanisms of tolerance is attributed to GABA_A receptors becoming downregulated (i.e. these receptors become less sensitive to GABA).^[4] When alcohol is no longer consumed, these down-regulated GABA_A receptor complexes are so insensitive to GABA that the typical amount of GABA produced has little effect, leading to physical withdrawal symptoms;^[4] since GABA normally inhibits neural firing, GABA_A receptor desensitization results in unopposed excitatory neurotransmission (i.e., fewer inhibitory postsynaptic potentialsoccur through GABA_A receptors), leading to neuronal over-excitation (i.e., more action potentials in the postsynaptic neuron). One of acamprosate’s mechanisms of action is the enhancement of GABA signaling at GABA_A receptors via positive allosteric receptor modulation.^[12][13] It has been purported to open the chloride ion channel in a novel way as it does not require GABA as a cofactor, making it less liable for dependence than benzodiazepines. Acamprosate has been successfully used to control tinnitus, hyperacusis, ear pain and inner ear pressure during alcohol use due to spasms of the tensor tympani muscle.^{[medical citation needed]}

In addition, alcohol also inhibits the activity of N-methyl-D-aspartate receptors (NMDARs).^[14][15] Chronic alcohol consumption leads to the overproduction (upregulation) of these receptors. Thereafter, sudden alcohol abstinence causes the excessive numbers of NMDARs to be more active than normal and to contribute to the symptoms of delirium tremensand excitotoxic neuronal death.^[16] Withdrawal from alcohol induces a surge in release of excitatory neurotransmitters like glutamate, which activates NMDARs.^[17] Acamprosate reduces this glutamate surge.^[18] The drug also protects cultured cells from excitotoxicity induced by ethanol withdrawal^[19] and from glutamate exposure combined with ethanol withdrawal.^[20]

Pharmacokinetics

Acamprosate is not metabolized by the human body.^[13] Acamprosate’s absolute bioavailability from oral administration is approximately 11%.^[13] Following administration and absorption of acamprosate, it is excreted unchanged (i.e., as acamprosate) via the kidneys.^[13]

History

Acamprosate was developed by Lipha, a subsidiary of Merck KGaA.^[21] and was approved for marketing in Europe in 1989.^{[citation needed]}

In October 2001 Forest Laboratories acquired the rights to market the drug in the US.^[21][22]

It was approved by the FDA in July 2004.^[23]

The first generic versions of acamprosate were launched in the US in 2013.^[24]

As of 2015 acamprosate was in development by Confluence Pharmaceuticals as a potential treatment for fragile X syndrome. The drug was granted orphan status for this use by the FDA in 2013 and by the EMA in 2014.^[25]

Society and culture

“Acamprosate” is the INN and BAN for this substance. “Acamprosate calcium” is the USAN and JAN. It is also technically known as N-acetylhomotaurine or as calcium acetylhomotaurinate.

It is sold under the brand name Campral.^[1]

Research

In addition to its apparent ability to help patients refrain from drinking, some evidence suggests that acamprosate is neuroprotective (that is, it protects neurons from damage and death caused by the effects of alcohol withdrawal, and possibly other causes of neurotoxicity).^[18][26]

References

 Acamprosate calcium

Substances Referenced in Synthesis Path

CAS-RN	Formula	Chemical Name	CAS Index Name
3687-18-1	C3H9NO3S	3-aminopropane-1-sulfonic acid	1-Propanesulfonic acid, 3-amino-
156-87-6	C3H9NO	3-amino-1-propanol	1-Propanol, 3-amino-

Trade Names

Country	Trade Name	Vendor	Annotation
D	Campral	Merck
F	Aotal	Merck Lipha
GB	Campral EC	Merck Serono
USA	Campral	Forest

Formulations

• tabl. 50 mg, 100 mg, 333 mg

References

• • DE 3 019 350 (Lab. Meram; appl. 21.5.1980; F-prior. 23.5.1979).
  • US 4 355 043 (Lab. Meram; 19.10.1982; F-prior. 23.5.1979).

• synthesis of 3-aminopropane-1-sulfonic acid:

  • Fujii, A. et al.: J. Med. Chem. (JMCMAR) 18, 502 (1975).
  • JP 46 002 012 (Kowa; appl. 19.1.1971).
  • WO 8 400 958 (Mitsui; appl. 15.3.1984; J-prior. 7.9.1982, 19.7.1983, 8.9.1982).

Acamprosate

Clinical data
Trade names	Campral EC
Synonyms	N-Acetyl homotaurine, Acamprosate calcium (JAN JP), Acamprosate calcium (USANUS)
Pregnancy category	C[1]> (US) B2 (AU)
Routes of administration	Oral [1]
ATC code	N07BB03 (WHO)
Legal status
Legal status	AU: S4 (Prescription only) UK: POM (Prescription only) US: ℞-only
Pharmacokinetic data
Bioavailability	11%[1]
Protein binding	Negligible[1]
Metabolism	Nil[1]
Elimination half-life	20 h to 33 h[1]
Excretion	Renal[1]
Identifiers
IUPAC name[show]
CAS Number	77337-76-9
PubChem CID	155434
IUPHAR/BPS	7106
DrugBank	DB00659
ChemSpider	136929
UNII	N4K14YGM3J
KEGG	D07058
ChEBI	CHEBI:51042
ChEMBL	CHEMBL1201293
ECHA InfoCard	100.071.495
Chemical and physical data
Formula	C5H11NO4S
Molar mass	181.211 g/mol
3D model (JSmol)	Interactive image
SMILES[hide] [Ca+2].O=C(NCCCS(=O)(=O)[O-])C.[O-]S(=O)(=O)CCCNC(=O)C
InChI[hide] InChI=1S/2C5H11NO4S.Ca/c2*1-5(7)6-3-2-4-11(8,9)10;/h2*2-4H2,1H3,(H,6,7)(H,8,9,10);/q;;+2/p-2  Key:BUVGWDNTAWHSKI-UHFFFAOYSA-L
(what is this?)  (verify)

////////////////Acamprosate calcium, アカンプロセート

CC(=O)NCCCS(O)(=O)=O

CC(=O)NCCCS(=O)(=O)[O-].CC(=O)NCCCS(=O)(=O)[O-].[Ca+2]

WATCH OUT FOR CORRECT FORMATTING TEXT AND STRUCTURE BOTH IN 2-3 DAYS

FDA approves the first drug with an indication for treatment of smallpox

 

Tecovirimat

4-trifluoromethyl-N-(3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop(f)isoindol-2(1H)-yl)-benzamide

N- [(3aR,4R,4aR,5aS,6S, 6aS)- 3,3a,4,4a,5,5a,6,6a- octahydro-1,3-dioxo- 4,6- ethenocycloprop[f]iso- indol-2(1H)-yl]-4- (trifluoromethyl)- benzamide

4 -trifluoromethyl -N- (3, 3a, 4, 4a, 5, 5a, 6, 6a- octahydro-1, 3 -dioxo-4, 6 -ethenocycloprop [f] isoindol -2 ( 1H) -yl ) – benzamide

Details

NDA FILED IN  US

2006 ORPHAN DRUG DESIGNATION IN US FOR SMALL POX

2010 ORPHAN DRUG DESIGNATION IN US FOR ORTHOPOX VIRUS

• N-((3aR,4R,4aR,5aS,6S,6aS)-1,3-Dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-ethenocyclopropa(f)isoindol-2(1H)-yl)-4-(trifluoromethyl)benzamide
• N-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-ethenocyclopropa[f]isoindol-2(1H)-yl)-4-(trifluoromethyl)benzamide

A core protein cysteine protease inhibitor potentially for treatment of smallpox infection.

SIGA TECHNOLOGIES INNOVATOR
SIGA-246; ST-246

CAS No. 869572-92-9

C19H15F3N2O3,

376.32921 g/mol

The Orthopox genus (Orthopoxyiridae) is a member of the Poxyiridae family and the Choropoxivirinae subfamily. The genus consists of numerous viruses that cause significant disease in human and animal populations. Viruses in the orthopox genus include cowpox, monkeypox, vaccina, and variola (smallpox), all of which can infect humans.

The smallpox (variola) virus is of particular importance. Recent concerns over the use of smallpox virus as a biological weapon has underscored the necessity of developing small molecule therapeutics that target orthopoxviruses. Variola virus is highly transmissible and causes severe disease in humans resulting in high mortality rates (Henderson et al. (1999) JAMA. 281:2127-2137). Moreover, there is precedent for use of variola virus as a biological weapon. During the French and Indian wars (1754-1765), British soldiers distributed blankets used by smallpox patients to American Indians in order to establish epidemics (Stern, E. W. and Stern A. E. 1945. The effect of smallpox on the destiny of the Amerindian. Boston). The resulting outbreaks caused 50% mortality in some Indian tribes (Stern, E. W. and Stern A. E.). More recently, the soviet government launched a program to produce highly virulent weaponized forms of variola in aerosolized suspensions (Henderson, supra). Of more concern is the observation that recombinant forms of poxvirus have been developed that have the potential of causing disease in vaccinated animals (Jackson et al. (2001) J. Virol., 75:1205-1210).

The smallpox vaccine program was terminated in 1972; thus, many individuals are no longer immune to smallpox infection. Even vaccinated individuals may no longer be fully protected, especially against highly virulent or recombinant strains of virus (Downie and McCarthy. (1958) J. Hyg. 56:479-487; Jackson, supra). Therefore, mortality rates would be high if variola virus were reintroduced into the human population either deliberately or accidentally.

Variola virus is naturally transmitted via aerosolized droplets to the respiratory mucosa where replication in lymph tissue produces asymptomatic infection that lasts 1-3 days. Virus is disseminated through the lymph to the skin where replication in the small dermal blood vessels and subsequent infection and lysis of adjacent epidermal cells produces skin lesions (Moss, B. (1990) Poxyiridae and Their Replication, 2079-2111. In B. N. Fields and D. M. Knipe (eds.), Fields Virology. Raven Press, Ltd., New York). Two forms of disease are associated with variola virus infection; variola major, the most common form of disease, which produces a 30% mortality rate and variola minor, which is less prevalent and rarely leads to death (<1%). Mortality is the result of disseminated intravascular coagulation, hypotension, and cardiovascular collapse, that can be exacerbated by clotting defects in the rare hemorrhagic type of smallpox (Moss, supra).

A recent outbreak of monkeypox virus underscores the need for developing small molecule therapeutics that target viruses in the orthpox genus. Appearance of monkeypox in the US represents an emerging infection. Monkeypox and smallpox cause similar diseases in humans, however mortality for monkeypox is lower (1%).

Vaccination is the current means for preventing orthopox virus disease, particularly smallpox disease. The smallpox vaccine was developed using attenuated strains of vaccinia virus that replicate locally and provide protective immunity against variola virus in greater than 95% of vaccinated individuals (Modlin (2001) MMWR (Morb Mort Wkly Rep) 50:1-25). Adverse advents associated with vaccination occur frequently (1:5000) and include generalized vaccinia and inadvertent transfer of vaccinia from the vaccination site. More serious complications such as encephalitis occur at a rate of 1:300,000, which is often fatal (Modlin, supra). The risk of adverse events is even more pronounced in immunocompromised individuals (Engler et al. (2002) J Allergy Clin Immunol. 110:357-365). Thus, vaccination is contraindicated for people with AIDS or allergic skin diseases (Engler et al.). While protective immunity lasts for many years, the antibody response to smallpox vaccination is significantly reduced 10 to 15 years post inoculation (Downie, supra). In addition, vaccination may not be protective against recombinant forms of ortho poxvirus. A recent study showed that recombinant forms of mousepox virus that express IL-4 cause death in vaccinated mice (Jackson, supra). Given the side effects associated with vaccination, contraindication of immunocompromised individuals, and inability to protect against recombinant strains of virus, better preventatives and/or new therapeutics for treatment of smallpox virus infection are needed.

Vaccinia virus immunoglobulin (VIG) has been used for the treatment of post-vaccination complications. VIG is an isotonic sterile solution of immunoglobulin fraction of plasma derived from individuals who received the vaccinia virus vaccine. It is used to treat eczema vaccinatum and some forms of progressive vaccinia. Since this product is available in limited quantities and difficult to obtain, it has not been indicated for use in the event of a generalized smallpox outbreak (Modlin, supra).

Cidofovir ([(S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine][HPMPC]) is a nucleoside analog approved for treatment of CMV retinitis in AIDS patients. Cidofovir has been shown to have activity in vitro against a number of DNA containing viruses including adenovirus, herpesviruses, hepadnaviruses, polyomaviruses, papillomaviruses, and ortho poxviruses (Bronson et al. (1990) Adv. Exp. Med. Biol. 278:277-83; De Clercq et al. (1987) Antiviral Res. 8:261-272; de Oliveira et al. (1996) Antiviral Res. 31:165-172; Snoeck et al. (2001) Clin Infect. Dis. 33:597-602). Cidofovir has also been found to inhibit authentic variola virus replication (Smee et al. (2002) Antimicrob. Agents Chemother. 46:1329-1335).

However, cidofovir administration is associated with a number of issues. Cidofovir is poorly bioavailable and must be administered intravenously (Lalezari et al. (1997) Ann. Intern. Med. 126:257-263). Moreover, cidofovir produces dose-limiting nephrotoxicity upon intravenous administration (Lalezari et al.). In addition, cidofovir-resistance has been noted for multiple viruses. Cidofovir-resistant cowpox, monkeypox, vaccinia, and camelpox virus variants have been isolated in the laboratory by repeated passage in the presence of drug (Smee, supra). Cidofovir-resistance represents a significant limitation for use of this compound to treat orthopoxvirus replication. Thus, the poor bioavailability, need for intravenous administration, and prevalence of resistant virus underscores the need for development of additional and alternative therapies to treat orthopoxvirus infection

In addition to viral polymerase inhibitors such as cidofovir, a number of other compounds have been reported to inhibit orthopoxvirus replication (De Clercq. (2001) Clin Microbiol. Rev. 14:382-397). Historically, methisazone, the prototypical thiosemicarbazone, has been used in the prophylactic treatment of smallpox infections (Bauer et al. (1969) Am. J. Epidemiol. 90:130-145). However, this compound class has not garnered much attention since the eradication of smallpox due to generally unacceptable side effects such as severe nausea and vomiting. Mechanism of action studies suggest that methisazone interferes with translation of L genes (De Clercq (2001), supra). Like cidofovir, methisazone is a relatively non-specific antiviral compound and can inhibit a number of other viruses including adenoviruses, picornaviruses, reoviruses, arboviruses, and myxoviruses (Id.).

Another class of compounds potentially useful for the treatment of poxviruses is represented by inhibitors of S-adenosylhomocysteine hydrolase (SAH). This enzyme is responsible for the conversion of S-adenosylhomocysteine to adenosine and homocysteine, a necessary step in the methylation and maturation of viral mRNA. Inhibitors of this enzyme have shown efficacy at inhibiting vaccinia virus in vitro and in vivo (De Clercq et al. (1998) Nucleosides Nucleotides. 17:625-634.). Structurally, all active inhibitors reported to date are analogues of the nucleoside adenosine. Many are carbocyclic derivatives, exemplified by Neplanacin A and 3-Deazaneplanacin A. While these compounds have shown some efficacy in animal models, like many nucleoside analogues, they suffer from general toxicity and/or poor pharmacokinetic properties (Coulombe et al. (1995) Eur. J. Drug Metab Pharmacokinet. 20:197-202; Obara et al. (1996) J. Med. Chem. 39:3847-3852). It is unlikely that these compounds can be administered orally, and it is currently unclear whether they can act prophylactically against smallpox infections. Identification of non-nucleoside inhibitors of SAH hydrolase, and other chemically tractable variola virus genome targets that are orally bioavailable and possess desirable pharmicokinetic (PK) and absorption, distribution, metabolism, elimination (ADME) properties would be a significant improvement over the reported nucleoside analogues. In summary, currently available compounds that inhibit smallpox virus replication are generally non-specific and suffer from use limiting toxicities and/or questionable efficacies.

In U.S. Pat. No. 6,433,016 (Aug. 13, 2002) and U.S. Application Publication 2002/0193443 A1 (published Dec. 19, 2002) a series of imidodisulfamide derivatives are described as being useful for orthopox virus infections.

Synthesis

RAW MATERIAL

Key RM is, 4,6-Etheno-1H-cycloprop[f]isobenzofuran-1,3(3aH)-dione, 3a,4,4a,5,5a,6-hexahydro-, (3aR,4R,4aR,5aS,6S,6aS)-rel–

cas  944-41-2, [US7655688]

Molecular Formula:	C11H10O3
Molecular Weight:	190.1953 g/mol

• 4,6-Etheno-1H-cycloprop[f]isobenzofuran-1,3(3aH)-dione, 4,4a,5,5a,6,6a-hexahydro-, (3aα,4β,4aα,5aα,6β,6aα)-
• Tricyclo[3.2.2.0^2,4]non-8-ene-6,7-dicarboxylic anhydride, stereoisomer (8CI)
• 3,6-Cyclopropylene-Δ⁴-tetrahydrophthalic anhydride

MP 94-96 °C

Ref, Dong, Ming-xin; European Journal of Medicinal Chemistry 2010, V45(9), Pg 4096-4103

SMILES……….

O=C1OC(=O)[C@H]4[C@@H]1[C@H]3C=C[C@@H]4[C@@H]2C[C@@H]23

SYNTHESIS CONTINUED…….

ST-246

Patent

WO2014028545

 

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=B0F96581458D847AE213FF5A89020C1D.wapp1nA?docId=WO2014028545&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

 

 

The present invention provides a process for making ST-246 outlined in Scheme 1

P = Boc

Scheme 1

The present invention also provides a process for making ST-246 outlined in, Scheme 2

Scheme 2

The present invention further provides a process for making ST-246 outlined in Scheme 3

ST-246

P = Boc

Scheme 3

P = Boc

Scheme 4

The present invention further provides a process for making ST-246 outlined in

Scheme 5

Scheme 5

 

Example 1 : Synthetic Route I:

P = Boc

Scheme 1

Step A. Synthesis of Compound 6 (P = Boc)

To a mixture of compound 3 (5.0 g, 26.3 mmol, synthesized according to WO041 12718) in EtOH (80 mL, EMD, AX0441 -3) was added terf-butyl carbazate 5 (3.65 g, 27.6 mmol, Aldrich, 98%). The reaction mixture was heated to reflux for 4 h under nitrogen atmosphere. LC-MS analysis of the reaction mixture showed less than 5% of compound 3 remained. The reaction mixture was evaporated under reduced pressure. The residue was recrystallized from EtOAc – hexanes, the solid was filtered, washed with hexanes (50 mL) and dried under vacuum to afford compound 6 (3.1 g, 39% yield) as a white solid. The filtrate was concentrated and purified by column chromatography eluting with 25% EtOAc in hexanes to give an additional 3.64 g (46% yield) of compound 6 as a white solid. Total yield: 6.74 g (84% yield). ¹H NMR in CDCI₃: δ 6.30 (br s, 1 H), 5.79 (t, 2H), 3.43 (s, 2H), 3.04 (s, 2H), 1 .46 (s, 9H), 1 .06-1 .16 (m, 2H), 0.18-0.36 (m, 2H); Mass Spec: 327.2 (M+Na)⁺

Step B. Synthesis of Compound 7 (HCI salt)

Compound 6 (3.6 g, 1 1 .83 mmol) was dissolved in /^‘-PrOAc (65 mL, Aldrich, 99.6%). 4M HCI in dioxane (10.4 mL, 41 .4 mmol, Aldrich) was added drop-wise to the above solution keeping the temperature below 20 °C. The reaction mixture was stirred at room temperature overnight (18 h) under nitrogen atmosphere. The resulting solid was filtered, washed with /^‘-PrOAc (15 mL) and dried under vacuum to yield HCI salt of compound 7 (1 .9 g, 67% yield) as a white solid. The filtrate was concentrated to 1/3 its volume and stirred at 10 – 15 °C for 30 min. The solid was filtered, washed with minimal volume of /^‘-PrOAc and dried to afford additional 0.6 g (21 % yield) of compound 7. Total yield: 2.5 g (88% yield). ¹ H NMR in DMSO-d6: δ 6.72 (br s, 3H), 5.68 (m, 2H), 3.20 (s, 2H), 3.01 (s, 2H), 1 .07-1 .17 (m, 2H), 0.18-0.29 (m, 1 H), -0.01 -0.07 (m, 1 H); Mass Spec: 205.1 (M+H)⁺

Step C. Synthesis of ST-246

To a mixture of compound 7 (0.96 g, 4 mmol) in dry dichloromethane (19 mL) was added triethylamine (1 .17 mL, 8.4 mmol, Aldrich) keeping the temperature below 20 °C. The resulting solution was stirred for 5 minutes at 15 – 20 °C, to it was added drop-wise 4-(trifluoromethyl)benzoyl chloride 8 (0.63 mL, 4.2 mmol, Aldrich, 97%) and the reaction mixture was stirred at room temperature overnight (18 h). LC-MS and TLC analysis showed the correct molecular weight and R_f value of ST-246 but the reaction was not complete. Additional 0.3 mL (2 mmol, 0.5 eq) of 4-(trifluoromethyl)benzoyl chloride 8 was added to the reaction mixture at 15 – 20 °C. The reaction was then stirred at room temperature overnight (19 h). LC-MS analysis indicated ca. 5% of starting material 7 still remained. The reaction was stopped and dichloromethane (30 mL) was added. The organic phase was washed with water (30 mL), saturated aqueous NH CI (30 mL), water (15 mL) and saturated aqueous NaHCO3 (30 mL). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated to give crude product. The crude product was purified by column chromatography eluting with 30 -50% EtOAc in hexanes to afford ST-246 (0.34 g, 23% yield) as an off-white solid. Analytical data (¹H NMR, LC-MS and HPLC by co-injection) were matched with those of ST-246 synthesized according to WO041 12718 and were consistent.

Example 2: Synthetic Route II

Scheme 2

Step A. Synthesis of Compound 9

A mixture of compound 4 (2.0 g, 9.8 mmol) and maleic anhydride 2 (0.96 g, 9.8 mmol, Aldrich powder, 95%) in o-xylene (100 mL, Aldrich anhydrous, 97%) was heated to reflux using a Dean-Stark trap apparatus overnight. After 18 h, LC-MS analysis at 215 nm showed the desired product 9 (86%), an uncyclized product (2.6%) and a dimer by-product (1 1 .6%).

Uncyclized product (MS = 303) Dimer by-product (MS = 489)

The reaction mixture was cooled to 45 °C and evaporated under reduced pressure. The residue was dissolved in EtOAc (50 mL) and the insoluble solid (mostly uncyclized product) was removed by filtration. The filtrate was concentrated and purified by column chromatography eluting with 50% EtOAc in hexanes to yield compound 9 (1 .5 g, 54% yield) as an off-white solid. ¹ H NMR in CDCI₃: δ 8.44 (s, 1 H), 7.91 (d, 2H), 7.68 (d, 2H), 6.88 (s, 2H); Mass Spec: 285.1 (M+H)⁺

Step B. Synthesis of ST-246 (Route II)

A mixture of compound 9 (0.97 g, 3.4 mmol) and cycloheptatriene 1 (0.51 mL, 4.42 mmol, distilled before use, Aldrich tech 90%) in toluene (50 mL, Aldrich anhydrous) was heated at 95 °C under nitrogen atmosphere. After 1 .5 h at 95 °C, LC-MS analysis at 254 nm showed 29% conversion to the desired product (endo:exo = 94:6). The resulting solution was continued to be heated at same temperature overnight. After 18 h at 95 °C, LC-MS analysis indicated 75% conversion with an endo:exo ratio of 94:6. The reaction temperature was increased to 1 10 °C and the reaction was monitored. After heating at 1 10 °C for 7 h, LC-MS analysis at 254 nm showed 96.4% conversion to the desired product (endo:exo = 94:6). The volatiles were removed by evaporation under reduced pressure and the reside was purified by column chromatography eluting with 30% EtOAc in hexanes to afford ST-246 (0.29 g, 22.6% yield, HPLC area 99.7% pure and 100% endo isomer) as a white solid. Analytical data (¹H NMR, LC-MS and HPLC by co-injection) were matched with those of ST-246 synthesized according to WO041 12718 and were consistent. An additional 0.5 g of ST-246 (38.9% yield, endo:exo = 97: 3) was recovered from column chromatography. Total Yield: 0.84 g (65.4% yield). ¹H NMR of ST-246 exo isomer in CDCI₃: δ 8.62 (s, 1 H), 7.92 (d, 2H), 7.68 (d, 2H), 5.96 (m, 2H), 3.43 (s, 2H), 2.88 (s, 2H), 1 .17 (s, 2H), 0.24 (q, 1 H), 0.13 (m, 1 H); Mass Spec: 377.1 (M+H)⁺

Example 3: Synthetic Route III

ST-246 9

P = Boc

Scheme 3

Step A. Synthesis of Compound 10

A mixture of maleic anhydride 2 (15.2 g, 155 mmol, Aldrich powder 95%) and terf-butyl carbazate 5 (20.5 g, 155 mmol, Aldrich, 98%) in anhydrous toluene (150 mL, Aldrich anhydrous) was heated to reflux using a Dean-Stark trap apparatus under nitrogen atmosphere. After refluxing for 2 h, no starting material 2 remained and LC-MS analysis at 254 nm showed the desired product 10 (20% by HPLC area), imine byproduct (18%) and disubstituted by-product (56%). The reaction mixture was concentrated and purified by column chromatography eluting with 25% EtOAc in hexanes to afford compound 10 (5.98 g, 18% yield, HPLC area >99.5% pure) as a white solid. ¹ H NMR in DMSO-d6: δ 9.61 (s, 1 H), 7.16 (s, 2H), 1 .42 (s, 9H); Mass Spec: 235.1 (M+Na)⁺.

duct

C₉H₁₂N₂0₄ C₁₄H₂₂N₄05

Mol. Wt.: 212.2 Mol. Wt.: 326.35

Step B. Synthesis of Compound 11 (HCI salt)

Compound 10 (3.82 g, 18 mmol) was dissolved in /^‘-PrOAc (57 mL, Aldrich, 99.6%). 4M HCI in dioxane (15.8 mL, 63 mmol, Aldrich) was added drop-wise to the above solution keeping the temperature below 20 °C. The solution was stirred overnight (24 h) at room temperature under nitrogen atmosphere. The resulting solid was filtered, washed with /^‘-PrOAc (10 mL) and dried at 45 °C under vacuum for 1 h to afford HCI salt of compound 11 (2.39 g, 89% yield) as a white solid. ¹ H NMR in CD₃OD: δ 6.98 (s, 2H); Mass Spec: 1 13.0 (M+H)⁺

Step C. Synthesis of Compound 9 (Route III)

To a mixture of compound 11 (1 .19 g, 8 mmol) in dry dichloromethane (24 mL) was added diisopropylethylannine (2.93 mL, 16.8 mmol, Aldrich redistilled grade) keeping the temperature below 20 °C. The resulting solution was stirred for 5 minute at 15 – 20 °C and to it was added 4-(trifluoromethyl)benzoyl chloride 8 (1 .31 mL, 8.8 mmol, Aldrich, 97%) drop-wise. The reaction was stirred at room temperature for 5 h. LC-MS analysis showed the correct MW but the reaction was not complete. Additional 0.48 mL (0.4 equiv) of 4-(trifluoromethyl)benzoyl chloride 8 was added to the reaction mixture at 15 – 20 °C and the reaction mixture was stirred at room temperature overnight (21 h). The reaction was stopped and dichloromethane (50 mL) was added. The organic phase was washed with water (50 mL), saturated aqueous NH₄CI (50 mL), water (30 mL) and saturated aqueous NaHCO3 (30 mL). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated to give crude product. The crude product was purified by column chromatography eluting with 30 – 35% EtOAc in hexanes to afford compound 9 (0.8 g, 35% yield) as a light pink solid. Analytical data (¹H NMR and LC-MS) were consistent with those of compound 9 obtained in Synthetic Route II.

Step D. Synthesis of ST-246 (Route III)

A mixture of compound 9 (0.5 g, 1 .76 mmol) and cycloheptatriene 1 (0.33 mL, 3.17 mmol, distilled before to use, Aldrich tech 90%) in toluene (10 mL, Aldrich anhydrous) was heated at 1 10 – 1 15 °C under nitrogen atmosphere. After 6 h, LC-MS analysis at 254 nm showed 95% conversion to the desired product (endo:exo = 94:6). The resulting solution was heated at same temperature overnight (22 h). LC-MS analysis at 254 nm showed no starting material 9 remained and the desired product (endo:exo = 93:7). The reaction mixture was concentrated and purified by column chromatography eluting with 25 – 35% EtOAc in hexanes to afford ST-246 (0.39 g, HPLC area >99.5% pure with a ratio of endo:exo = 99:1 ) as a white solid. Analytical data (¹ H NMR, LC-MS and HPLC by co-injection) were compared with those of ST-246 synthesized according to WO041 12718 and were found to be consistent. An additional 0.18 g of ST-246 (HPLC area >99.5% pure, endo:exo = 91 : 9) was recovered from column chromatography. Total Yield: 0.57 g (86% yield).

Example 4 ; Synthetic Route IV:

P = Boc

Scheme 4

Step A. Synthesis of Compound 10

A mixture of maleic anhydride 2 (3.4 g, 34.67 mmol, Aldrich powder, 95%) and terf-butyl carbazate 5 (4.6 g, 34.67 mmol, Aldrich, 98%) in anhydrous toluene (51 ml_, Aldrich) was heated to reflux using a Dean-Stark trap apparatus under nitrogen atmosphere. After refluxing for 2.5 h, no starting material 2 remained and LC-MS analysis at 254 nm showed the desired product 10 (19% HPLC area), imine by-product (18%) and another by-product (56%). The reaction mixture was concentrated and purified by column chromatography eluting with 30% EtOAc in hexanes to afford compound 10 (1 .0 g, 13.6% yield, HPLC area >99% pure) as a white solid. Analytical data (¹H NMR and LC-MS) were consistent with those of compound 10 obtained in Synthetic Route III.

Im ine by-product

Mol. Wt.: 212.2

Step B. Synthesis of Compound 6

A mixture of compound 10 (4.4 g, 20.74 mmol) and cycloheptatriene 1 (3.22 mL, 31 .1 mmol, distilled before to use, Aldrich tech 90%) in toluene (88 mL, 20 volume, Aldrich anhydrous) was heated at 95 °C under nitrogen atmosphere. After 15 h at 95 °C, LC-MS analysis showed 83% conversion to the desired product. The reaction mixture was heated at 105 °C overnight. After total 40 h at 95 – 105 °C, LC-MS analysis at 254 nm showed -99% conversion to the desired product (endo:exo = 93:7). The reaction mixture was concentrated and the crude was purified by column chromatography eluting with 25 – 50 % EtOAc in hexanes to afford compound 6 (2.06 g, 32.6% yield, HPLC area 99.9% pure and 100% endo isomer) as a white solid. ¹ H NMR and LC-MS were consistent with those of compound 6 obtained in Synthetic Route I. An additional 4.0 g of 6 (63.4% yield, HPLC area 93% pure with a ratio of endo:exo = 91 : 9) was recovered from column chromatography. Total Yield: 6.06 g (96% yield).

Step C. Synthesis of Compound 7 (HCI salt)

Compound 6 (2.05 g, 6.74 mmol) was dissolved in /^‘-PrOAc (26 mL, Aldrich, 99.6%). 4M HCI in dioxane (5.9 mL, 23.58 mmol, Aldrich) was added drop-wise to the above solution keeping the temperature below 20 °C. The solution was stirred overnight (18 h) at room temperature under nitrogen atmosphere. The resulting solid was filtered, washed with /^‘-PrOAc (5 mL) and dried under vacuum to yield HCI salt of compound 7 (1 .57 g, 97% yield) as a white solid. Analytical data (¹ H NMR and LC-MS) were consistent with those of compound 7 in Synthetic Route I.

Step D. Synthesis of ST-246 (Route IV)

To a mixture of compound 7 (0.84 g, 3.5 mmol) in dichloromethane (13 mL) was added diisopropylethylamine (1 .34 mL, 7.7 mmol) keeping the temperature below 20 °C and the resulting solution was stirred for 5 – 10 minutes. 4-(Trifluoromethyl)benzoyl chloride 8 (0.57 mL, 3.85 mmol, Aldrich, 97%) was added to above solution keeping the temperature below 20 °C. The reaction mixture was stirred at room temperature for 2 h. Additional 0.2 mL (0.4 equiv) of 4-(trifluoromethyl)benzoyl chloride 8 was added to the reaction keeping the temperature below 20 °C. The reaction was stirred at room temperature overnight (24 h). The reaction mixture was diluted with dichloromethane (20 mL). The organic phase was washed with water (20 mL), saturated aqueous NH₄CI (20 mL), water (20 mL) and saturated aqueous NaHCO3 (20 mL). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated to give crude product. The crude product was purified by column chromatography eluting with 30 – 35% EtOAc in hexanes to afford ST-246 (0.25 g, 19% yield, HPLC area >99.5% pure) as a white solid. Analytical data (¹H NMR and LC-MS) were consistent with those of ST-246 synthesized according to WO041 12718.

Example 5: Synthetic Route V:

Scheme 5

Step A. Synthesis of Compound 13

To a mixture of compound 7 (1 .6 g, 6.65 mmol, synthesized according to Synthetic Route I) in dichloromethane (80 ml_,) was added triethylamine (2.04 ml_, 14.63 mmol) keeping the temperature below 20 °C and the resulting solution was stirred for 5 – 10 minute. 4-lodobenzoyl chloride 12 (1 .95 g, 7.31 mmol, 1 .1 equiv, Aldrich) was added portion-wise under nitrogen atmosphere to the above solution keeping the temperature below 20 °C. The reaction mixture was stirred at room temperature overnight. After 17 h and 19 h, additional 0.35 g (0.2 equiv) of acid chloride 12 was added to the reaction keeping the temperature below 20 °C. After 24 h, additional 0.18 g (0.1 equiv, used total 1 .6 equiv) of acid chloride 12 was added and the reaction was continued to stir at room temperature overnight (total 43 h). LC-MS analysis at 215 nm showed 43% of the desired product (13) and -5% of compound 7. The reaction was diluted with dichloromethane (100 ml_). The organic phase was washed with saturated aqueous NH₄CI (100 ml_), water (100 ml_) and saturated aqueous NaHCO₃ (100 ml_). The organic phase was separated, dried over Na2SO₄, filtered and concentrated to give crude product. The crude product was purified by column chromatography eluting with 25 – 50% EtOAc in hexanes to afford compound 13 (1 .63 g, 57% yield, HPLC area 93% pure) as a white solid. ¹ H NMR in DMSO-d6: δ 1 1 .19 and 10.93 (two singlets with integration ratio of 1 .73:1 , total of 1 H, same proton of two rotamers), 7.93 (d, 2H), 7.66 (d, 2H), 5.80 (s, 2H), 3.36 (s, 2H), 3.27 (s, 2H), 1 .18 (s, 2H), 0.27 (q, 1 H), 0.06 (s,1 H); Mass Spec: 435.0 (M+H)⁺

Step B. Synthesis of ST-246 (Route V)

Anhydrous DMF (6 ml_) was added to a mixture of compound 13 (0.2 g, 0.46 mmol), methyl 2, 2-difluoro-2-(fluorosulfonyl)acetate (0.44 ml_, 3.45 mmol, Aldrich) and copper (I) iodide (90 mg, 0.47 mmol). The reaction mixture was stirred at -90 °C for 4 h. LC-MS analysis at 254 nm indicated no starting material 13 remained and showed 48% HPLC area of ST-246. The reaction mixture was cooled to 45 °C and DMF was removed under reduced pressure. The residue was slurried in EtOAc (30 mL) and insoluble solid was removed by filtration. The filtrate was concentrated and purified by column chromatography eluting with 25 – 35% EtOAc in hexanes to afford ST-246 (55

mg, 32% yield, 95% pure by HPLC at 254 nm) as off-white solid. Analytical data (¹H NMR and LC-MS) were consistent with those of ST-246 synthesized according to WO041 12718.

PAPER

N-(3,3a,4,4a,5,5a,6,6a-Octahydro-1,3-dioxo-4,6- ethenocycloprop[f]isoindol-2-(1H)-yl)carboxamides:  Identification of Novel Orthopoxvirus Egress Inhibitors

Thomas R. Bailey ,*† Susan R. Rippin ,† Elizabeth Opsitnick ,† Christopher J. Burns ,† Daniel C. Pevear ,† Marc S. Collett ,† Gerry Rhodes ,† Sanjeev Tohan ,† John W. Huggins ,‡ Robert O. Baker ,‡§ Earl R. Kern ,‖ Kathy A. Keith ,‖Dongcheng Dai ,⊥ Guang Yang ,⊥ Dennis Hruby ,⊥ and Robert Jordan ⊥

ViroPharma Incorporated, 397 Eagleview Boulevard, Exton, Pennsylvania 19341, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Frederick, Maryland 21702, University of Alabama, Birmingham, Alabama 35294, and SIGA Technologies, Inc., 4575 SW Research Way, Corvallis, Oregon 97333

J. Med. Chem., 2007, 50 (7), pp 1442–1444

DOI: 10.1021/jm061484y

A series of novel, potent orthopoxvirus egress inhibitors was identified during high-throughput screening of the ViroPharma small molecule collection. Using structure−activity relationship information inferred from early hits, several compounds were synthesized, and compound 14was identified as a potent, orally bioavailable first-in-class inhibitor of orthopoxvirus egress from infected cells. Compound 14 has shown comparable efficaciousness in three murine orthopoxvirus models and has entered Phase I clinical trials.

http://pubs.acs.org/doi/suppl/10.1021/jm061484y

http://pubs.acs.org/doi/suppl/10.1021/jm061484y/suppl_file/jm061484ysi20070204_060607.pdf

General Procedure for synthesis of compounds 2-14, 16-18.

N-(3,3a,4,4a,5,5a,6,6aoctahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl)-4- (trifluoromethyl)benzamide (14).

A mixture of 2.00 g (9.8 mmol) of 4-(trifluoromethyl) benzoic acid hydrazide, 1.86 g (9.8 mmol) of 4,4a,5,5a,6,6a-hexahydro-4,6-etheno-1Hcycloprop[f]isobenzofuran-1,3(3aH)-dione, and one drop of diisopropylethylamine in 40 mL of absolute ethanol was refluxed for 4.5 h. Upon cooling to rt, 4 mL of water was added, and the product began to crystallize. The suspension was cooled in an ice bath, and the precipitate collected by filtration. The crystalline solid was air-dried affording 3.20 g (87%) of the product as a white solid;

Mp 194-195 ºC. 1 H NMR, (300 MHz, d6 -DMSO) δ 11.20, 11.09 (2 brs from rotamers, 1H), 8.06 (d, J= 7.8 Hz, 2H), 7.90 (d, J= 7.8 Hz, 2H), 5.78 (m, 2H), 3.26 (m, 4H), 1.15 (m, 2H), 0.24 (dd, J= 7.2, 12.9 Hz, 1H), 0.04 (m, 1H).

Anal. calcd. for C19H15F3N2O3● 0.25H2O: %C, 59.92; %H, 4.10; %F, 14.97; %N, 7.36; %O, 13.65. Found: %C, 59.97; %H, 4.02; %F, 14.94; %N, 7.36; %O, 13.71.

CLICK ON IMAGE

PATENT

US20140316145

CLICK ON IMAGE

http://www.google.com/patents/US8802714

Example 1

Preparation of 4-trifluoromethyl-N-(3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl)-benzamide

a. Preparation of Compounds 1(a) and 1(b).

A mixture of cycloheptatriene (5 g, 54.26 mmol) and maleic anhydride (6.13 g, 62.40 mmol) in xylenes (35 mL) was heated at reflux under argon overnight. The reaction was cooled to room temperature and a tan precipitate was collected by filtration and dried to give 2.94 grams (28%) of the desired product, which is a mixture of compounds 1(a) and 1(b). Compound 1(a) is normally predominant in this mixture and is at least 80% by weight. The purity of Compound 1(a) may be further enhanced by recrystallization if necessary. Compound 1(b), an isomer of compound 1(a) is normally less than 20% by weight and varies depending on the conditions of the reaction. Pure Compound 1(b) was obtained by concentrating the mother liquid to dryness and then subjecting the residue to column chromatography. Further purification can be carried out by recrystallization if necessary. ¹H NMR (500 MHz) in CDCl₃: δ 5.95 (m, 2H), 3.42 (m, 2H), 3.09 (m, 2H), 1.12 (m, 2H), 0.22 (m, 1H), 0.14 (m, 1H).

b. Preparation of N-[(3aR,4R,4aR,5aS,6S,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl]-4-(trifluoromethyl)-benzamide. desired

A mixture of compound 1(a) (150 mg, 0.788 mmol) and 4-trifluoromethylbenzhydrazide (169 mg, 0.827 mmol) in ethanol (10 mL) was heated under argon overnight. The solvent was removed by rotary evaporation. Purification by column chromatography on silica gel using 1/1 hexane/ethyl acetate provided 152 mg (51%) of the product as a white solid.

c. Preparation of N-[(3aR,4S,4aS,5aR,6R,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl]-4-(trifluoromethyl)-benzamide. UNWANTED

N-[(3aR,4S,4aS,5aR,6R,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl]4-(trifluoromethyl)-benzamide was prepared and purified in the same fashion as for N-[(3aR,4R,4aR,5aS,6S,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl]-4-(trifluoromethyl)-benzamide by replacing 1(a) with 1(b) and was obtained as a white solid. ¹H NMR (300 MHz) in CDCl₃: δ 8.62 (s, 1H), 7.92 (d, 2H), 7.68 (d, 2H), 5.96 (m, 2H), 3.43 (s, 2H), 2.88 (s, 2H), 1.17 (s, 2H), 0.24 (q, 1H), 0.13 (m, 1H); Mass Spec: 377.1 (M+H)⁺.

FINAL COMPD SYNTHESIS

TABLE 1
Example			**Mass
Number	R6	*NMR	Spec	Name
1		1H NMR in DMSO-d6: δ 11.35 (d, 1H); 11.09 (d, 1H); 8.08 (d, 2H); 7.92 (d, 2H); 5.799 (s, 2H); 3.29 (brs, 4H); 1.17 (m, 2H); 0.26 (m, 1H); 0.078 (s, 1H)	375 (M − H)−	N-[(3aR,4R,4aR,5aS,6S, 6aS)-3,3a,4,4a,5,5a,6,6a- octahydro-1,3-dioxo- 4,6-ethenocycloprop[f] isoindol-2(1H)-yl]-4- (trifluoromethyl)- benzamide

TABLE 1 EXAMPLE 1

N- [(3aR,4R,4aR,5aS,6S, 6aS)- 3,3a,4,4a,5,5a,6,6a- octahydro-1,3-dioxo- 4,6- ethenocycloprop[f]iso- indol-2(1H)-yl]-4- (trifluoromethyl)- benzamide

¹H NMR in DMSO-d₆: δ 11.35 (d, 1H); 11.09 (d, 1H); 8.08 (d, 2H); 7.92 (d, 2H); 5.799 (s, 2H); 3.29 (brs, 4H); 1.17 (m, 2H); 0.26 (m, 1H); 0.078 (s, 1H), 375 (M − H)⁻

EXAMPLE 42 Characterization of 4-trifluoromethyl-N-(3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl)-benzamide (“ ”)

In the present application, ST-246 refers to: N-[(3aR,4R,4aR,5aS,65,6aS)-3,3a,4,4a,5,5a,6,6a-octahydro-1,3-dioxo-4,6-ethenocycloprop[f]isoindol-2(1H)-yl]-4-(trifluoromethyl)-benzamide.

Physico-Chemical Properties

Appearance: ST-246 is a white to off-white powder.

Melting Point: Approximately 196° C. by DSC.

Permeability: The calculated log P is 2.94. Based on the partition coefficient, ST-246 is expected to have good permeability.

Particle Size: The drug substance is micronized to improve its dissolution in the gastrointestinal fluids. The typical particle size of the micronized material is 50% less than 5 microns.

Solubility: The solubility of ST-246 is low in water (0.026 mg/mL) and buffers of the gastric pH range. Surfactant increases its solubility slightly. ST-246 is very soluble in organic solvents. The solubility data are given in Table 5.

CLICK ON IMAGE

PATENT

http://www.google.com/patents/CN101445478A?cl=en

Tecovirimat (ST-246) is an antiviral with activity against orthopoxviruses such as smallpox and is currently undergoing clinical trials. It was previously owned by Viropharma and discovered in collaboration with scientists at USAMRIID. It is currently owned and is synthesized by Siga Technologies, a drug development company in the biodefense arena. It works by blocking cellular transmission of the virus, thus preventing the disease. Tecovirimat has been effective in laboratory testing, with no serious side effects reported to date. Despite not yet having FDA approval for medical use, tecovirimat is stockpiled in the US Strategic National Stockpile as a defense against a smallpox outbreak.^[1]

Clinical study

The results of clinical trials involving tecovirimat supports its use against smallpox and other related orthopoxviruses. It has shown potential for a variety of uses including prophylaxis, as a post-exposure therapeutic, as a therapeutic and an adjunct to vaccination.^[2]

Tecovirimat can be taken orally and has recently been granted permission to conduct Phase II trials by the U.S. Food and Drug Administration (FDA). In phase I trials tecovirimat was generally well tolerated with no serious adverse events.^[3] Due to its importance for biodefense, the FDA has designated tecovirimat for ‘fast-track’ status, creating a path for expedited FDA review and eventual regulatory approval.

Tecovirimat is an orthopoxvirus egress inhibitor. Tecovirimat appears to target the V061 gene in cowpox, which is homologous to the vaccinia virus F13L. By targeting this gene, tecovirimat inhibits the function of a major envelope protein required for the production of extracellar virus. Thus the virus is prevented from leaving the cell, and the spread of the virus within the body is prevented.^[4]

 

References

1. Damon, Inger K.; Damaso, Clarissa R.; McFadden, Grant (2014). “Are We There Yet? The Smallpox Research Agenda Using Variola Virus”. PLoS Pathogens 10 (5): e1004108.doi:10.1371/journal.ppat.1004108. PMID 24789223.
2. Siga Technologies
3. Jordan, R; Tien, D; Bolken, T. C.; Jones, K. F.; Tyavanagimatt, S. R.; Strasser, J; Frimm, A; Corrado, M. L.; Strome, P. G.; Hruby, D. E. (2008). “Single-Dose Safety and Pharmacokinetics of ST-246, a Novel Orthopoxvirus Egress Inhibitor”. Antimicrobial Agents and Chemotherapy 52 (5): 1721–1727. doi:10.1128/AAC.01303-07. PMC 2346641. PMID 18316519.
4. Yang, G; Pevear, D. C.; Davies, M. H.; Collett, M. S.; Bailey, T; Rippen, S; Barone, L; Burns, C; Rhodes, G; Tohan, S; Huggins, J. W.; Baker, R. O.; Buller, R. L.; Touchette, E; Waller, K; Schriewer, J; Neyts, J; Declercq, E; Jones, K; Hruby, D; Jordan, R (2005). “An Orally Bioavailable Antipoxvirus Compound (ST-246) Inhibits Extracellular Virus Formation and Protects Mice from Lethal Orthopoxvirus Challenge”. Journal of Virology 79 (20): 13139–13149. doi:10.1128/JVI.79.20.13139-13149.2005. PMC 1235851. PMID 16189015.

Referenced by
Citing Patent Filing date Publication date Applicant Title
CN101912389A * Aug 9, 2010 Dec 15, 2010 中国人民解放军军事医学科学院微生物流行病研究所 Pharmaceutical composition containing ST-246 and preparation method and application thereof
CN102406617A * Nov 30, 2011 Apr 11, 2012 中国人民解放军军事医学科学院生物工程研究所 Tecovirimat dry suspension and preparation method thereof
CN102406617B Nov 30, 2011 Aug 28, 2013 中国人民解放军军事医学科学院生物工程研究所 Tecovirimat dry suspension and preparation method thereof
CN103068232B * Mar 23, 2011 Aug 26, 2015 西佳科技股份有限公司 多晶型物形式st-246和制备方法
US8530509 Jul 29, 2011 Sep 10, 2013 Siga Technologies, Inc. Compounds, compositions and methods for treatment and prevention of orthopoxvirus infections and associated diseases
US8802714 Aug 14, 2013 Aug 12, 2014 Siga Technologies, Inc. Compounds, compositions and methods for treatment and prevention of orthopoxvirus infections and associated diseases
US9045418 Jul 3, 2014 Jun 2, 2015 Siga Technologies, Inc. Compounds, compositions and methods for treatment and prevention of Orthopoxvirus infections and associated diseases

Patent Citations

Cited Patent	Filing date	Publication date	Applicant	Title
US20070287735 *	Apr 23, 2007	Dec 13, 2007	Siga Technologies, Inc.	Chemicals, compositions, and methods for treatment and prevention of orthopoxvirus infections and associated diseases
US20090011037 *	Apr 23, 2008	Jan 8, 2009	Cydex Pharmaceuticals, Inc.	Sulfoalkyl Ether Cyclodextrin Compositions and Methods of Preparation Thereof
Citing Patent	Filing date	Publication date	Applicant	Title
US8530509	Jul 29, 2011	Sep 10, 2013	Siga Technologies, Inc.	Compounds, compositions and methods for treatment and prevention of orthopoxvirus infections and associated diseases
US8802714	Aug 14, 2013	Aug 12, 2014	Siga Technologies, Inc.	Compounds, compositions and methods for treatment and prevention of orthopoxvirus infections and associated diseases
US9045418	Jul 3, 2014	Jun 2, 2015	Siga Technologies, Inc.	Compounds, compositions and methods for treatment and prevention of Orthopoxvirus infections and associated diseases

Classifications

Tecovirimat

Systematic (IUPAC) name
N-{3,5-Dioxo-4- azatetracyclo[5.3.2.0{2,6}.0{8,10}]dodec-11-en-4- yl}-4-(trifluoromethyl)benzamide
Identifiers
UNII	F925RR824R
ChEMBL	CHEMBL1242629
Synonyms	ST-246
Chemical data
Formula	C19H15F3N2O3
Molecular mass	base: 376.3 g/mol

//////////////////Tecovirimat, FDA 2018, ORPHAN DRUG DESIGNATION,  TPOXX, SIGA Technologies Inc,  Fast Track, Priority Review

FC(F)(F)c1ccc(cc1)C(=O)NN1C(=O)C2C(C3C=CC2C2CC32)C1=O

DR ANTHONY MEVIN CRASTO Conferred prestigious individual national award at function for contribution to Pharma society from Times Network, National Awards for Marketing Excellence ( For Excellence in HEALTHCARE) | 5th July, 2018 | Taj Lands End, Mumbai India

////////////National award,  contribution to Pharma society, Times Network, Excellence in HEALTHCARE,  5th July, 2018, Taj Lands End, Mumbai,  India, ANTHONY CRASTO

#hotpersoninawheelchair
#worlddrugtracker

Mercaptamine bitartrate

2-aminoethanethiol;2,3-dihydroxybutanedioic acid

Cystagon; Cysteamine – Mylan/Orphan Europe; Cysteamine bitartrate

Procysbi; CYSTEAMINE BITARTRATE; 27761-19-9; CHEBI:50386; (+/-)-Tartaric Acid

• OriginatorMylan
• DeveloperAlphapharm; Mylan
• ClassMercaptoethylamines; Small molecules; Sulfhydryl compounds
• Mechanism of ActionGlutathione synthase stimulants

Highest Development Phases

• MarketedNephropathic cystinosis
• DiscontinuedUnspecified

Most Recent Events

• 09 Apr 2018Mercaptamine bitartrate licensed to Recordati worldwide
• 26 Oct 2017Chemical structure information added
• 31 Dec 2008Mercaptamine bitartrate oral is still in phase II/III trials for Undefined indication in European Union

DESCRIPTION: CYSTAGON® (cysteamine bitartrate) Capsules for oral administration, contain cysteamine bitartrate, a cystine depleting agent which lowers the cystine content of cells in patients with cystinosis, an inherited defect of lysosomal transport. CYSTAGON® is the bitartrate salt of cysteamine, an aminothiol, beta-mercaptoethylamine. Cysteamine bitartrate is a highly water soluble white powder with a molecular weight of 227 and the molecular formula C2H7NS · C4H6O6. It has the following chemical structure:

Cysteamine is a medication intended for a number of indications, and approved by the FDA to treat cystinosis.

It is stable aminothiol, i.e., an organic compound containing both an amine and a thiol functional groups. Cysteamine is a white, water-soluble solid. It is often used as salts of the ammonium derivative [HSCH₂CH₂NH₃]^+[1] including the hydrochloride, phosphocysteamine, and bitartrate.^[2]

Cysteamine molecule is biosynthesized in mammals, including humans, by the degradation of coenzyme A. The intermedia pantetheineis broken down into cysteamine and pantothenic acid.^[2] It is the biosynthetic precursor to the neurotransmitter hypotaurine.^[3][4]

Medical uses

Cysteamine is used to treat cystinosis. It is available by mouth (capsule and extended release capsule) and in eye drops.^{[5][6][7][8][9]}

Adverse effects

Topical use

The most important adverse effect related to topical use might be skin irritation.

Oral use

The label for oral formulations of cysteamine carry warnings about symptoms similar to Ehlers-Danlos syndrome, severe skin rashes, ulcers or bleeding in the stomach and intestines, central nervous symptoms including seizures, lethargy, somnolence, depression, and encephalopathy, low white blood cell levels, elevated alkaline phosphatase, and idiopathic intracranial hypertension that can cause headache, tinnitus, dizziness, nausea, double or blurry vision, loss of vision, and pain behind the eye or pain with eye movement.^[6]

The main side effects are Ehlers-Danlos syndrome, severe skin rashes, ulcers or bleeding in the stomach and intestines, central nervous symptoms, low white blood cell levels, elevated alkaline phosphatase, and idiopathic intracranial hypertension (IIH). IIH can cause headache, ringing in the ears, dizziness, nausea, blurry vision, loss of vision, and pain behind the eye or with eye movement.

Additional adverse effects of oral cysteamine include bad breath, skin odor, vomiting, nausea, stomach pain, diarrhea, and loss of appetite.^[6]

The drug is in pregnancy category C; the risks of cysteamine to a fetus are not known but it harms babies in animal models at doses less than those given to people.^[7][8]

For eye drops, the most common adverse effects are sensitivity to light, redness, and eye pain, headache, and visual field defects.^[8]

Interactions

There are no drug interactions for normal capsules or eye drops,^[7][8] but the extended release capsules should not be taken with drugs that affect stomach acid like proton pump inhibitors or with alcohol, as they can cause the drug to be released too quickly.^[6] It doesn’t inhibit any cytochrome P450 enzymes.^[6]

Pharmacology

People with cystinosis lack a functioning transporter (cystinosin) which transports cystine from the lysosome to the cytosol. This ultimately leads to buildup of cystine in lysosomes, where it crystallizes and damages cells.^[5] Cysteamine enters lysosomes and converts cystine into cysteine and cysteine-cysteamine mixed disulfide, both of which can exit the lysosome.^[6]

Biological function

Cysteamine also promotes the transport of L-cysteine into cells, that can be further used to synthesize glutathione, which is one of the most potent intracellular antioxidants.^[4]

Cysteamine is used as a drug for the treatment of cystinosis; it removes cystine that builds up in cells of people with the disease.^[10]

History

First evidence regarding the therapeutic effect of cysteamine on cystinosis dates back to 1950s. Cysteamine was first approved as a drug for cystinosis in the US in 1994.^[6] An extended release form was approved in 2013.^[11]

Society and culture

It is approved by FDA and EMA.^[5][6]

In 2013, the regular capsule of cysteamine cost about $8,000 per year; the extended release form that was introduced that year was priced at $250,000 per year.^[11]

Research

It was studied in in vitro and animal models for radiation protection in the 1950s, and in similar models from the 1970s onwards for sickle cell anemia, effects on growth, its ability to modulate the immune system, and as a possible inhibitor of HIV.^[2]

In the 1970s it was tested in clinical trials for Paracetamol toxicity which it failed, and in clinical trials for systemic lupus erythematosus in the 1990s and early 2000s, which it also failed.^[2]