Methods and processes for preparing MCT4 inhibitors

Abstract

Processes for preparing heterocyclic compounds and compositions for use as MCT4 inhibitors are provided.

Description

[Technical Field]

[0001] Disclosed herein are processes for preparing heterocyclic compounds and compositions for use as MCT4 inhibitors. [Background technology]

[0002] Lactate transport from glycolytic cells is normally mediated by the monocarboxylate transporter MCT4, which has a weak affinity for lactate (K m = 28 mM), which, in combination with a high turnover rate, allows for the rapid transport of large amounts of lactate. MCT4 expression is typically restricted to highly glycolytic tissues, such as white muscle fibers, lymphocytes, astrocytes, and Sertoli cells. While MCT4 is absent in most normal tissues, in many cancer indications, including colorectal cancer, glioma, head and neck cancer, triple-negative breast cancer, prostate cancer, KRAS-mutated lung cancer, liver cancer, and kidney cancer, MCT4 expression is highly upregulated and correlates with reduced survival.

[0003] The correlation between MCT4 expression and poor cancer outcomes appears to be related to important functional consequences in multiple cancer models. Stable expression of MCT4 is highly tumorigenic in respiratory-impaired, Ras-transformed fibroblast xenograft models. Conversely, silencing MCT4 slows or eliminates tumor growth in xenograft models of breast cancer, colon cancer, and glioma. In xenograft models of breast cancer and colon cancer, MCT4 expression is required for inflammatory cytokine IL-8-mediated angiogenesis. MCT4 has also been shown to play an important role in various aspects of cancer cell migration, invasion, and the Warburg effect (e.g., growth on glucose, extracellular acidification, and lactate secretion).

[0004] Inhibition of MCT4-mediated lactate transport may be an effective strategy to reduce the Warburg effect in cancer. Unfortunately, no potent and selective MCT4 inhibitors have been reported. Moderate to weak MCT4 inhibitors are known (e.g., phloretin and α-CN-4-OH-cinnamate). However, these compounds indiscriminately inhibit several other transporters, including MCT1.

[0005] Novel potent inhibitors of MCT4 are described, for example, in WO 2016 / 201426, the contents of which are incorporated herein by reference in their entirety. The compound 2-([1-[2-(azetidin-1-yl)phenyl]-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl]methoxy)-2-methylpropanoic acid and related compounds are described in WO 2018 / 111904, the contents of which are incorporated herein by reference in their entirety, as inhibitors of MCT4 with promising potential. Summary of the Invention [Problem to be solved by the invention]

[0006] There is a need for new and improved methods for the synthesis of 2-([1-[2-(azetidin-1-yl)phenyl]-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl]methoxy)-2-methylpropanoic acid and related compounds that are amenable to large-scale synthesis.

[0007] Throughout this application, the citation of any reference should not be construed as an admission that such reference is prior art to the present application. [Means for solving the problem]

[0008] Formula I [ka] or a salt thereof, which comprises: i) Formula II [ka] with a compound of formula III [ka] in a polar aprotic solvent in the presence of a non-nucleophilic base to form a compound of formula IV [ka] and forming a compound of formula (I): ii) hydrolyzing the compound of formula IV to produce a compound of formula I or a salt thereof; wherein R 1 is a C1-C4 alkyl, and R 2 is a halogen.

[0009] prepared by the processes described herein, [ka] Also provided is a compound of the formula: wherein the compound contains a detectable amount of one or more organic solvents that is less than 3% by weight.

[0010] Furthermore, the formula [ka] or a salt thereof.

[0011] Furthermore, the formula [ka] or a salt thereof.

[0012] Also provided herein is a method for treating a monocarboxylic acid transporter MCT4-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound described herein or a salt thereof, or a composition described herein.

[0013] These and other aspects of the invention disclosed herein will be explained in more detail as the patent disclosure proceeds. DETAILED DESCRIPTION OF THE INVENTION

[0014] Formula I [ka] or a salt thereof, which comprises: i) Formula II [ka] with a compound of formula III [ka] in a polar aprotic solvent in the presence of a non-nucleophilic base to form a compound of formula IV [ka] and forming a compound of formula (I): ii) hydrolyzing the compound of formula IV to produce a compound of formula I or a salt thereof; wherein R 1 is a C1-C4 alkyl, and R 2 is a halogen.

[0015] In some embodiments, R 1 is isopropyl, and R 2 is bromine.

[0016] In some embodiments, the molar ratio of the compound of Formula III to the compound of Formula II is from about 7.0 to about 10.0.

[0017] In some embodiments, the non-nucleophilic base is selected from N,N-diisopropylethylamine (DIPEA), 8-diazabicycloundec-7-ene (DBU), 1,5-diazabicyclo(4.3.0)non-5-ene (DBN), 2,6-di-tert-butylpyridine, tert-butyllithium, tert-butylphosphazene, lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), potassium tert-butoxide, potassium bis(trimethylsilyl)amide (KHMDS), lithium tetramethylpiperidide (LiTMP), sodium hydride, potassium hydride, and sodium tert-butoxide. In some embodiments, the non-nucleophilic base is potassium bis(trimethylsilyl)amide (KHMDS).

[0018] In some embodiments, the molar ratio of KHMDS to the compound of Formula II is from about 4.0 to about 6.0.

[0019] In some embodiments, the polar aprotic solvent is selected from acetone, acetonitrile, dichloromethane, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethyl acetate, hexamethylphosphoric triamide (HMPT), pyridine, tetrahydrofuran (THF), and mixtures thereof.

[0020] In some embodiments, the polar aprotic solvent is a mixture composed of THF and DMF.

[0021] In some embodiments, hydrolyzing the compound of Formula IV comprises reacting the compound of Formula IV with a nucleophilic base.

[0022] In some embodiments, the nucleophilic base is selected from sodium hydroxide and sodium methoxide.

[0023] In some embodiments, the molar ratio of the nucleophilic base to the compound of Formula IV is from about 18.0 to about 20.0.

[0024] In some embodiments, the compound of formula II has formula V: [ka] (In the formula, R 3 is a halogen) is prepared by contacting the compound of formula (I) with azetidine in the presence of a palladium catalyst.

[0025] In some embodiments, R 3 is bromine.

[0026] In some embodiments, the molar ratio of azetidine to the compound of Formula V is from about 2.0 to about 3.0.

[0027] In some embodiments, the molar ratio of the palladium catalyst to the compound of Formula V is from about 0.05 to about 0.15.

[0028] In some embodiments, the palladium catalyst is selected from Pd(OAc), palladium(II) pivalate, tetrakis(triphenylphosphine)palladium(0), bis(acetonitrile)palladium(II) dichloride, bis(triphenylphosphine)palladium(II) dichloride, [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, tris(dibenzylideneacetone)dipalladium(0), and palladium(II) chloride.

[0029] In some embodiments, the palladium catalyst is Pd(OAc) 2 .

[0030] In some embodiments, the contacting is carried out in the presence of a ligand.

[0031] In some embodiments, the ligand is trimethylphosphine, triphenylphosphine, tricyclohexylphosphine, tri(o-tolyl)phosphine, 2-(dicyclohexylphosphino)-2',4',6'-tri-i-propyl-1,1'-biphenyl (XPhos), 2-(dicyclohexylphosphino)-2'-(N,N-dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl, 2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl, butyldi-1-adamantylphosphine, 2-(di-t-butylphosphino)biphenyl, 2-(dicyclohexylphosphino)biphenyl, (R)-(-)-1-[(S )-2-(diphenylphosphino)ferrocenyl]ethyldicyclohexylphosphine, 1,2-bis(diphenylphosphino)benzene (dppbenzene), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), bis(2-diphenylphosphinophenyl)ether (DPEphos), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos), 1,4-bis(diphenylphosphino)butane (dppb), 1,2-bis(diphenylphosphino)ethane (dppe), 1,1'-bis(diphenylphosphino)ferrocene (dppf), 1,3-bis(diphenylphosphino)propane (dppp), and [(t-Bu3)PH]BF4.

[0032] In some embodiments, the ligand is xantphos.

[0033] In some embodiments, the molar ratio of the ligand to the compound of formula V is from about 0.05 to about 0.15.

[0034] In some embodiments, the compound of formula V has formula VI [ka] with a compound of formula VII: [ka] by contacting a compound of formula VIII: [ka] and thereafter contacting a compound of formula VIII with a reducing agent to obtain a compound of formula V, wherein R 3 is a halogen.

[0035] In some embodiments, the reducing agent is lithium aluminum hydride, sodium bis(2-methoxyethoxy)aluminum hydride (Red-Al), nascent hydrogen, sodium amalgam, zinc amalgam, sodium borohydride, lithium borohydride, SmI, Fe 2+ Compounds containing ions, Sn 2+ The compound is selected from compounds containing ions, hydrazine, diisobutylaluminum hydride, and any combination thereof.

[0036] In some embodiments, the reducing agent is sodium borohydride.

[0037] In some embodiments, the compound of formula VI is prepared by reacting 1-(3-hydroxyphenyl)ethan-1-one with a compound of formula IX: [ka] (In the formula, R 4 is selected from halogens) in a polar aprotic solvent in the presence of a non-nucleophilic base to form 1-(3-cyclobutoxyphenyl)ethan-1-one, which is then contacted with dimethyl oxalate and a strong base to provide the compound of formula VII.

[0038] In some embodiments, the polar aprotic solvent is selected from acetone, acetonitrile, dichloromethane, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethyl acetate, hexamethylphosphoric triamide (HMPT), pyridine, tetrahydrofuran (THF), and mixtures thereof.

[0039] In some embodiments, the non-nucleophilic base is selected from cesium carbonate, sodium carbonate, and potassium carbonate.

[0040] In some embodiments, the strong base is selected from sodium hydroxide, sodium methoxide, sodium ethoxide, lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), potassium tert-butoxide, potassium bis(trimethylsilyl)amide (KHMDS), lithium tetramethylpiperidide (LiTMP), sodium hydride, potassium hydride, and sodium tert-butoxide.

[0041] In some embodiments, the molar ratio of non-nucleophilic base to 1-(3-hydroxyphenyl)ethan-1-one is from about 1.0 to about 2.0.

[0042] In some embodiments, the process comprises reacting a compound of formula I with a compound of formula [ka] The method further comprises converting the compound to a Tris salt of

[0043] Also provided are Tris salts prepared by the processes described herein.

[0044] Additionally, compounds of the formula [ka] Also provided are compounds of the formula: which contain detectable amounts, in some embodiments less than 3% by weight, of residual organic solvent.

[0045] at least 90%, for example at least 95%, 96%, 97%, 98%, 99% or 99.5% of a compound of formula I or a salt thereof and a detectable amount of a compound of formula IV, a compound of formula II, a compound of formula V, a compound of formula VIII, 1-(3-hydroxyphenyl)ethan-1-one, bromocyclobutane, cesium carbonate, hydrochloric acid, 2-bromophenylhydrazine, sodium borohydride, Xantphos, sodium tert-butoxide, isopropyl 2-bromo-2-methylpropionate, potassium bis(trimethylsilyl)amide, sodium hydroxide, water Also provided is a composition comprising one or more impurities selected from the group consisting of sodium borohydride, sodium tert-pentoxide, acetonitrile, dichloromethane, 4-dimethylaminopyridine, ethyl acetate, dimethylformamide, ethanol, water, isopropanol, isopropyl acetate, potassium bis(trimethylsilyl)amide, methanol, methyl tert-butyl ether, sodium borohydride, N-methyl-2-pyrrolidone, palladium diacetate, sodium triacetoxyborohydride, tetrahydrofuran, and one or more impurities selected from one or more heavy metals.

[0046] In some embodiments, the one or more heavy metals are selected from platinum, palladium, iridium, rhodium, rhenium, ruthenium, cadmium, mercury, lead, arsenic, manganese, chromium, cobalt, nickel, copper, zinc, selenium, silver, antimony, thallium, nickel, vanadium, and zinc.

[0047] Also provided herein is a method of treating a monocarboxylate transporter MCT4-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound described herein or a composition described herein.

[0048] In some embodiments, the monocarboxylate transporter MCT4-mediated disorder is cardiac hypertrophy.

[0049] In some embodiments, the monocarboxylate transporter MCT4-mediated disorder is heart failure.

[0050] In some embodiments, the monocarboxylate transporter MCT4-mediated disorder is an MCT4-expressing cancer.

[0051] In some embodiments, the monocarboxylate transporter MCT4-mediated disorder is rheumatoid arthritis.

[0052] formula [ka] or a salt thereof.

[0053] Furthermore, the formula [ka] or a salt thereof.

[0054] To facilitate understanding of this disclosure, some terms and abbreviations used herein are defined below.

[0055] When introducing elements of the disclosure or preferred embodiments thereof, the articles "a," "an," "the," and "said" are intended to mean that there are one or more of the elements. The terms "comprise," "include," and "have" are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0056] When used in a list of two or more items, the term "and / or" means that any one of the listed items may be used by itself or in combination with any one or more of the listed items. For example, the phrase "A and / or B" is intended to mean either or both of A and B, i.e., A alone, B alone, or A and B in combination. The phrase "A, B, and / or C" is intended to mean A alone, B alone, C alone, A and B in combination, A and C in combination, B and C in combination, or A, B, and C in combination.

[0057] As used herein, the term "about" when referring to a measurable value such as compound amount, dosage, time, temperature, etc., is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5% or even 0.1% from the specified amount.

[0058] When a range of values is disclosed and the notation "n1... to n2" or "between n1... and n2" is used, where n1 and n2 are numbers, unless otherwise specified, this notation is intended to include the numbers themselves and the range therebetween. The range may be an integer or continuous value therebetween, and includes the end values. For example, a range of "2 to 6 carbon atoms" is intended to include carbon numbers 2, 3, 4, 5, and 6, since the number of carbon atoms is an integer unit. For example, "1 to 3 μM (micromolar)" is intended to include 1 μM, 3 μM, and all significant figures therebetween (e.g., 1.255 μM, 2.1 μM, 2.9999 μM, etc.).

[0059] The term "alkyl," as used herein, alone or in combination, refers to a straight- or branched-chain alkyl group containing 1 to 20 carbon atoms. In certain embodiments, alkyl contains 1 to 10 carbon atoms. In yet other embodiments, alkyl contains 1 to 6 carbon atoms. Alkyl groups can be optionally substituted as defined herein. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl, nonyl, and the like. The term "alkylene," as used herein, alone or in combination, refers to a saturated aliphatic group derived from a straight- or branched-chain saturated hydrocarbon bonded at two or more positions, e.g., methylene (—CH—). Unless otherwise specified, the term "alkyl" can include "alkylene" groups.

[0060] The terms "halo" or "halogen," as used herein, alone or in combination, refer to fluorine, chlorine, bromine, or iodine.

[0061] As used herein, the term "non-nucleophilic base" refers to a sterically hindered organic base that is a weak nucleophile. Examples of non-nucleophilic bases include N,N-diisopropylethylamine (DIPEA), 8-diazabicycloundec-7-ene (DBU), 1,5-diazabicyclo(4.3.0)non-5-ene (DBN), 2,6-di-tert-butylpyridine, tert-butyllithium, tert-butylphosphazene, lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), potassium tert-butoxide, potassium bis(trimethylsilyl)amide (KHMDS), lithium tetramethylpiperidide (LiTMP), sodium hydride, potassium hydride, sodium tert-butoxide, and potassium tert-butoxide. In certain embodiments, the non-nucleophilic base is potassium bis(trimethylsilyl)amide (KHMDS).

[0062] As used herein, the term "nucleophilic base" refers to a Bronsted-Lowry base that is an excellent nucleophile. Examples include sodium methoxide, methyllithium, sodium hydroxide, lithium hydroxide, sodium cyanide, potassium cyanide, sodium acetylide, sodium amide, sodium iodide, lithium bromide, potassium iodide, and sodium azide.

[0063] As used herein, the term "polar solvent" refers to a solvent that has a large dipole moment.

[0064] As used herein, the term "polar aprotic solvent" refers to a polar solvent lacking acidic hydrogen. Therefore, they are not hydrogen bond donors. Examples of polar aprotic solvents include acetone, acetonitrile, dichloromethane, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethyl acetate, hexamethylphosphoric triamide (HMPT), pyridine, and tetrahydrofuran (THF). In certain embodiments, the polar aprotic solvent is THF, DMF, or a combination of THF and DMF.

[0065] The term "disease" as used herein is generally intended to be synonymous with, and used interchangeably with, the terms "disorder" and "condition" (in medical conditions), in that it refers to any abnormal condition of the human or animal body or one of its parts that impairs normal functioning, is typically manifested by characteristic signs and symptoms, and reduces the lifespan or quality of life of the human or animal.

[0066] The compounds disclosed herein may exist as therapeutically acceptable salts. The present invention includes the compounds described above in the form of salts, such as acid addition salts. Suitable salts include those formed with both organic and inorganic acids. Such acid addition salts are usually pharmaceutically acceptable. However, pharmaceutically unacceptable salts may be useful in the preparation and purification of the compounds. Basic addition salts may also be formed, which may also be pharmaceutically acceptable.

[0067] The term "therapeutically acceptable salt," as used herein, refers to a water- or oil-soluble or dispersible and therapeutically acceptable salt or zwitterionic form of a compound disclosed herein. These salts can be prepared during the final isolation and purification of the compound, or can be prepared separately by reacting the free base form of the appropriate compound with a suitable acid. Representative acid addition salts include acetate, adipate, alginate, L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, butyrate, camphorate, camphorsulfonate, citrate, digluconate, formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate (isethionate), lactate, maleate, and malonate. The salts include DL-mandelate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphonate, picrate, pivalate, propionate, pyroglutamate, succinate, sulfonate, tartrate, L-tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, para-toluenesulfonate (p-tosylate), and undecanoate. The basic groups of the compounds disclosed herein can also be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, bromides, and iodides; and benzyl and phenethyl bromides. Examples of acids that can be used to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid. Salts can also be formed by coordination of an alkali metal or alkaline earth ion with a compound. Thus, the present invention contemplates sodium, potassium, magnesium, calcium, and other salts of the compounds disclosed herein.

[0068] Base addition salts can be prepared during the final isolation and purification of the compounds by reacting the carboxyl group with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a metal cation, or with ammonia or an organic primary, secondary, or tertiary amine. Therapeutically acceptable salt cations include lithium, sodium, potassium, calcium, magnesium, and aluminum, as well as non-toxic quaternary amine cations such as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine, and N,N'-dibenzylethylenediamine. Other representative organic amines useful for the formation of base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine, and piperazine.

[0069] Salts of compounds can be prepared by reacting the free base form of the appropriate compound with an appropriate acid.

[0070] The compounds disclosed herein may exist as polymorphs and other discrete solid forms such as solvates, hydrates, etc. The compounds may be polymorphs, solvates, or hydrates of a salt or free base or acid.

[0071] In some embodiments, the process further comprises formulating the product and / or its salt into a pharmaceutical composition. The pharmaceutical composition may comprise one or more pharmaceutically acceptable carriers. The pharmaceutical composition may be administered orally. The pharmaceutical composition may be delivered orally using tablets, lozenges, liquids, emulsions, suspensions, drops, capsules, caplets, or gel caps, and other oral administration methods known to those skilled in the art. Suitable excipients for pharmaceutical compositions include one or more fillers, binders, and surfactants, glidants, lubricants, disintegrants, swelling agents, and antioxidants.

[0072] As used herein, the term "detectable" refers to a measurable amount as determined using an HPLC method with a detection limit of 0.01 area %.

[0073] As used herein, the term "not detectable" in reference to the amount of an impurity means not detected by the HPLC method described herein, which has a detection limit of 0.01 area % for the impurity.

[0074] As used herein, "limit of detection (LOD)" or "detection limit" refers to the lowest concentration of analyte that can be unambiguously detected above a baseline signal that is approximately three times the baseline signal noise.

[0075] Fillers include, but are not limited to, lactose, saccharose, glucose, starch, microcrystalline cellulose, fine cellulose, mannitol, sorbitol, calcium hydrogen phosphate, aluminum silicate, amorphous silica, and sodium chloride, starch, and calcium phosphate dibasic dihydrate. In one embodiment, the filler can absorb water but is not water-soluble. In one embodiment, the filler is a spheronization aid. The spheronization aid may include one or more of crospovidone, carrageenan, chitosan, pectic acid, glycerides, β-cyclodextrin (β-CD), cellulose derivatives, microcrystalline cellulose, powdered cellulose, polyplasdone crospovidone, and polyethylene oxide.

[0076] Binders include, but are not limited to, cellulose ethers, methylcellulose, ethylcellulose, hydroxyethylcellulose, propylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose (hypromellose, e.g., hypromellose 2910, Methocel™ E), carboxymethylcellulose, starch, pregelatinized starch, acacia, tragacanth, gelatin, polyvinylpyrrolidone (povidone), cross-linked polyvinylpyrrolidone, sodium alginate, microcrystalline cellulose, and lower alkyl-substituted hydroxypropylcellulose. In one embodiment, the binder is selected from wetting binders.

[0077] Surfactants include, but are not limited to, anionic surfactants such as sodium lauryl sulfate, sodium deoxycholate, dioctyl sodium sulfosuccinate, and sodium stearyl fumarate, nonionic surfactants such as polyoxyethylene ethers and polysorbate 80, and cationic surfactants including quaternary ammonium compounds. In one embodiment, the surfactant is selected from anionic surfactants, for example, sodium lauryl sulfate.

[0078] Disintegrants include, but are not limited to, starch, cross-linked sodium carboxymethylcellulose, carmellose sodium, carmellose calcium, cross-linked polyvinylpyrrolidone and sodium starch glycolate, low-substituted hydroxypropyl cellulose and hydroxypropyl starch.

[0079] Lubricants include, but are not limited to, polyethylene glycol of various molecular weights, magnesium stearate, calcium stearate, calcium silicate, fumed silicon dioxide, magnesium carbonate, magnesium lauryl sulfate, aluminum stearate, stearic acid, palmitic acid, cetyl alcohol, stearol, and talc.

[0080] Lubricants include, but are not limited to, stearic acid, magnesium stearate, calcium stearate, aluminum stearate, and siliconized talc.

[0081] In certain embodiments, the formulation further comprises one or more antioxidants. Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc., (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc., and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0082] method General synthetic methods for preparing compounds The present invention below is further illustrated by the examples set forth below.

[0083] In the examples below and throughout this disclosure, the following abbreviations may be used: DMF = dimethylformamide, KHMDS = potassium bis(trimethylsilyl)amide, MTBE = methyl tert-butyl ether, TBME = tert-butyl methyl ether, NMP = N-methyl-2-pyrrolidone, THF = tetrahydrofuran, 1 H-NMR = proton nuclear magnetic resonance, ICP = inductively coupled plasma mass spectrometry, IPC = in-process control / check. Other abbreviations may be used and will be familiar to those skilled in the art. [Example]

[0084] Example 1: Synthesis of 2-([1-[2-(azetidin-1-yl)phenyl]-5-(3-cyclobutoxyphenyl)-1H-pyrazol-3-yl]methoxy)-2-methylpropanoic acid 1. Synthesis of Compound 7 Compound 7 was synthesized by the process in Scheme 1. Scheme 1 [ka]

[0085] Step 1: [ka] A mixture of 1-(3-hydroxyphenyl)ethan-1-one (20 kg), bromocyclobutane (1.3 eq.), and CsCO (1.5 eq.) in DMF (5 vol.) was stirred at 80±5°C for 16 h. The reaction vessel was charged with water (15 vol.) and methyl tert-butyl ether (MTBE, 15 vol.). The organic layer was separated, washed twice with 20% brine (5 vol.), concentrated to approximately 5 vol., and then solvent exchanged three times with methanol to approximately 5 vol. The crude methanol solution containing compound 1 was used in the next step without further purification.

[0086] Step 2: [ka] To the crude methanol solution from Step 1 (5 volumes) was added sodium methoxide (2.0 eq.) and dimethyl oxalate (1.5 eq.). The reaction vessel was stirred at 30±5°C for 16 hours. Upon completion, the reaction mixture was cooled to a temperature of 0-10°C. The pH was adjusted to 2-3 with 4.0 M HCl in methanol, and the crude solution containing compound 2 was used in the next step without further purification.

[0087] Step 3: [ka] 2-Bromophenylhydrazine hydrochloride (1.0 eq.) was added to the reaction vessel containing the crude methanol solution from Step 2. The reaction mixture was then stirred at 60±5°C for 10 hours and then cooled to 5-15°C. The resulting solid was filtered, washed with methanol (1 vol.), and then slurried in water (20 vol.). The solid was filtered again and washed with water (2 vol.). The product was dried in an oven at 50°C to give 54.9 kg of compound 3 (88% yield for Steps 1-3).

[0088] Step 4: [ka] Compound 3 (4.0 kg, 1.0 eq.) was added to THF (4 vol.), followed by the addition of methanol (0.4 vol.) to the reaction vessel, which was then cooled to 10–25°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 10–25°C for 16 h. After completion of the reaction, the reaction was cooled to 0–10°C, and the pH was adjusted to 3–5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5–15°C for an additional 1–2 h. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 3.5 kg of compound 4 in 85% yield.

[0089] Step 4 was repeated on a 5x scale using compound 3 (20.0 kg, 1.0 eq.) in THF (6.0 vol.), followed by the addition of methanol (0.4 vol.) to the reaction vessel, which was then cooled to 25±5°C. Sodium borohydride (1.2 eq.) was added, and the reaction was stirred at 25±5°C for 16 hours. After completion of the reaction, the reaction was cooled to 0-10°C and the pH was adjusted to 3-5 with 0.5 M HCl, at which point the product precipitated from solution. The mixture was stirred at 5-15°C for an additional 1-2 hours. The product was collected by filtration, washed with water, and dried in an oven at 50°C to give 18.2 kg of compound 4 in 99.4% purity and 97% isolated yield.

[0090] Step 5: [ka] Compound 4 (2.97 kg, 1.0 eq.) was added to THF (10 vol.) under a nitrogen atmosphere with stirring. Xantphos (0.11 eq.), Pd(OAc) (0.11 eq.), and t-BuONa (2.0 eq.) were added to the reaction vessel. Azetidine (2.5 eq.) was added, and the resulting mixture was stirred at 25-35°C for 20 hours. MTBE (20 vol.) and water (20 vol.) were added to the vessel. The organic layer was separated, washed with 5% aqueous NH4Cl and 5% aqueous NaCl, and filtered through Celite®. The resulting organic layer was concentrated and solvent exchanged with ethyl acetate (10 vol.). The resulting solution was stirred at 70-80°C for 30-30 minutes, then cooled to 15-20°C and stirred for an additional 1-2 hours. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and dried in an oven at 50° C. to give 1.7 kg of compound 5 in 60% yield. Reproduced with 18.0 kg of compound 4, 10.8 kg of compound 5 was obtained in 98.8% purity and 63.8% isolated yield.

[0091] Step 6 [ka] To a solution of compound 5 (700.0 g, 1 eq.) in DMF (8 vol.) was added isopropyl 2-bromo-2-methylpropanoate (8.0 eq.). The reaction mixture was cooled to -60°C to -50°C using liquid nitrogen. A solution of potassium bis(trimethylsilyl)amide (KHMDS, 5.0 eq.) in THF was added dropwise over 1.5 h, and the reaction was stirred for 30–50 min. Water (10 vol.) was added to the reaction vessel. The organic phase was collected, washed twice with NaCl (15%) (5 vol.), and then concentrated. Crude compound 6 (70% yield) was used directly in the next step without further purification.

[0092] Step 7 [ka] Crude compound 6 (1950.0 g, 1 eq.) from Step 6 was added to a reaction vessel along with 30% (aq.) NaOH (10 vol, 20 eq.) and methanol (5 vol). The reaction was stirred at 50±5°C for 16 h and then cooled to 15–30°C. The solution was washed twice with MTBE (10 vol), after which the aqueous layer was separated and filtered through Celite®. The pH of the resulting aqueous phase was adjusted to 3–5 with 2.0 M HCl. The solution was cooled to 5–10°C and stirred for 2–3 h before filtering. The filter cake was dissolved in THF (10 vol), and then mercaptosilica gel (300 g, 15% w / w) was added to the solution. The resulting mixture was heated to 50–60°C, stirred for 3 h, and then filtered. The solution was concentrated, and the crude product was dissolved in ethyl acetate (10% v / v). The resulting solution was heated to 70–80° C. and stirred for 3–4 h, then cooled to 0–10° C. The solid was collected by filtration, washed with ethyl acetate (0.5 vol), and then dried in an oven at 50° C. to give compound 7 (1.4 kg, 58% yield) as an off-white solid.

[0093] This reaction was repeated on a larger scale by adding crude compound 6 (8.4 kg, 1 eq.) from step 6 to a reaction vessel along with 30% (aq.) KOH (10 vol, 20 eq.) and methanol (5 vol.). After performing the same workup as above, 8.76 kg of compound 7 was obtained over two steps in 97.7% purity and 67% yield.

[0094] 2. Use of various propanoic acid esters in Step 6 In step 6, various propanoic acid esters were investigated, and the results are shown in Table 1. [ka]

[0095] [Table 1]

[0096] [Table 2]

[0097] As shown in Table 1, starting materials using the isopropyl ester performed better than t-butyl and methyl esters. Furthermore, the use of the isopropyl ester limits the potential for the formation of dimer (or bis-adduct) by-products detected when the methyl ester was used in Run 13. The use of the isopropyl ester also allows for the use of solvents other than DMF / NaH. The reaction also proceeded much faster than when DMF / NaH was used.

[0098] 3. Formation of the Tris Salt of Compound 7 [ka] Compound 7 (8.4 kg, 1 eq.) was added to a reaction vessel containing THF (42 L, 5.0 vol.), charged with mercaptosilica gel (420 g, 5 wt.%), and stirred and heated at 50–60°C for 3 hours. The reaction mixture was sampled, concentrated, and subjected to ICP analysis to determine the residual Pd concentration (approximately 15 ppm). The reaction mixture was filtered, and acetone (168 L, 20.0 vol.) and aqueous tris(hydroxymethyl)aminomethane ("Tris", 1.01 eq.) solution (4.2 L, 0.5 vol.) were added. The reaction mixture was stirred at 20–30°C for 5–10 minutes to dissolve most of the solids. Stirring was continued at 20–30°C for approximately 20 hours to precipitate the Tris salt of compound 7. The precipitate was filtered, washed with acetone (4.2 L, 0.5 vol), and dried under vacuum at 60°C to give 8.4 kg of compound 7 tris containing 1690 ppm residual acetone and 962 ppm residual THF in 99.5% purity and 79.2% yield.

[0099] Example 2: Pharmacokinetic assay The Tris salt and the free acid of the compound of Formula I were tested in two animal models to evaluate pharmacokinetic parameters.

[0100] Rat model Groups, Dosing, and Collections. The pharmacokinetics of the free acid and Tris salt of the compound of Formula I by oral gavage were evaluated in male Sprague-Dawley rats. Three rats per group were orally administered 20, 60, or 200 mg of the free acid or 25, 75, or 250 mg of 0.5% methylcellulose in saline to final concentrations of 2, 6, or 20 mg / mL of the free acid or 2.5, 7.5, or 20 mg / mL. Plasma was collected from the jugular vein at 5, 15, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after administration. No abnormal clinical signs were observed.

[0101] Stock and Dose Preparation. Stock solutions were prepared by dissolving 2.47 mg of the compound of Formula I (free acid) in 2.470 mL of DMSO with vortexing to give a 1 mg / mL solution of the free acid, or 2.09 mg of the Tris salt in 1.655 mL of DMSO with vortexing to give a 1 mg / mL solution of the Tris salt. Dose solutions were prepared by vortexing / sonicating the following solids in the solvent:

[0102] [Table 3]

[0103] LC MS-MS Analysis. Liquid chromatography with tandem mass spectrometry was used to measure plasma concentrations of the compound of Formula I free acid and the compound of Formula I tris salt in plasma samples collected at predetermined time points.

[0104] Appropriate serial concentrations of working solutions were obtained by diluting the analyte stock solution with 50% acetonitrile in aqueous solution. Five µL of working solution (10, 20, 50, 100, 500, 1000, 5000, 8000, and 10,000 ng / mL) was added to 50 µL of blank male SD rat plasma to achieve calibration standards ranging from 1 to 1,000 ng / mL (1, 2, 5, 10, 50, 100, 500, 800, and 1000 ng / mL) in a total volume of 55 µL. Five quality control (QC) samples of plasma were prepared independently from those used for the calibration curve: 2 ng / mL, 5 ng / mL, 10 ng / mL, 50 ng / mL, and 800 ng / mL. These QC samples were prepared on the day of analysis using the same method as the calibration standards. 55 μL of standard, 55 μL of QC sample, and 55 μL of unknown sample (50 μL of plasma / blood and 5 μL of blank solution) were each added to 200 μL of methanol mixture containing internal standard (dexamethasone) for protein precipitation and vortexed for 30 seconds. After centrifugation at 4°C and 4000 rpm for 15 minutes, the supernatant was diluted 1:2 with water. 2 μL of the supernatant was injected into the LC / MS / MS system for quantitative analysis.

[0105] The instrumentation included a HALO 90A C18 2.7 μm 2.1 × 50 mm HPLC column, a Prominence degasser DGU-20A5R(C), a Shimadzu LC-30AD liquid chromatograph with a communication bus module CBM-20A and an Auto SIL-20AC HT, and an AB Sciex Triple Quad 5500 LC / MS / MS system. The following conditions were used:

[0106] [Table 4]

[0107] Results. Results are shown in Table 3 (free acid) and Table 4 (Tris salt). Note that the Tris salt (FW = 582.7) has a higher formula weight than the free acid (MW = 461.6), so the appropriate comparison is 25 mg / kg Tris salt versus 20 mg / kg free acid. Overall, the total drug exposure (AUC lastand AUC Inf ) was higher in the Tris salt group than in the free acid group in both male and female subjects. max ) was also higher in the Tris salt group than in the free acid group in both males and females, except for females given 20 / 25 mg / kg, where the free acid C max (78,883ng / mL) in Tris C max (68,567 ng / mL). In general, exposure was higher in female than in male subjects.

[0108] [Table 5]

[0109] [Table 6]

[0110] Dog Model Groups, Dosing, and Collections. The pharmacokinetics of the free acid and Tris salt of the compound of Formula I administered by oral capsule were evaluated in dogs. Three male beagle dogs were administered a single dose of 3 mg / kg of the free acid on study day 1 and a single dose of 3 mg / kg of the Tris salt on study day 8. Plasma was collected by peripheral venipuncture pre-dose and at 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. No abnormal clinical signs were observed.

[0111] In a follow-up study, the pharmacokinetics of the Tris salt of the compound of Formula I was examined by oral gavage of a 0.5% methylcellulose formulation in saline. Two groups of animals were studied. In Group 1, three male beagle dogs were administered a single dose of 3 mg / kg of the Tris salt. In Group 2, three male beagle dogs were administered a single dose of 30 mg / kg of the Tris salt. Plasma was collected by peripheral venipuncture before administration and at 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. No abnormal clinical signs were observed.

[0112] LC MS-MS Analysis. Liquid chromatography with tandem mass spectrometry was used to measure plasma concentrations of the compound of Formula I free acid and the compound of Formula I tris salt in plasma samples collected at predetermined time points. 18 The instruments included a Shimadzu LC-30AD liquid chromatograph with an S-5 μm (50 × 2.1 mm) HPLC column, a CBM-20A communication bus module, and an Auto SIL-20AC HT, and a Triple Quad 5500 LC / MS / MS system. Tolbutamide was used as the internal standard. The following conditions were used:

[0113] [Table 7]

[0114] [Table 8]

[0115] Results. Results from the powder capsule formulation are shown in Table 5. Results from oral gavage of the suspension are shown in Table 6. Overall, total drug exposure (AUC last and AUC Inf ) was higher for the Tris salt (both powder capsule and suspension formulations) compared to the free acid. The half-life appeared to be shorter for the Tris salt compared to the free acid, but the mean residence time was increased. Because one animal in the salt powder capsule formulation experiment had abnormally high plasma concentrations (30-50 times higher than the other two subjects) at the first two time points, SD and CV% were not calculated.

[0116] [Table 9]

[0117] [Table 10]

[0118] Other embodiments The foregoing detailed description is provided to aid those skilled in the art in practicing the present disclosure. However, the disclosure described and claimed herein is not limited in scope by the specific embodiments disclosed herein, as these embodiments are intended as illustrations of certain aspects of the present disclosure. Any equivalent embodiments are intended to be within the scope of the present disclosure. Indeed, various modifications of the present disclosure in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description, without departing from the spirit or scope of the inventive findings. Such modifications are also intended to be encompassed within the scope of the appended claims.

Claims

1. Formula I 【Chemical 1】 or a salt thereof, comprising: i) Formula II 【Chemistry 2】 with a compound of formula III 【Chemistry 3】 in a polar aprotic solvent in the presence of a non-nucleophilic base to produce a compound of formula IV 【Chemistry 4】 and forming a compound of formula (I): ii) hydrolyzing the compound of formula IV to produce the compound of formula I or a salt thereof; wherein R １ is C １ ~C ４ is alkyl, and R ２ is a halogen, process.

2. R １ is isopropyl, and R ２ 2. The process of claim 1, wherein is bromine.

3. 10. The process of claim 1, wherein the molar ratio of the compound of formula III to the compound of formula II is from about 7.0 to about 10.

0.

4. 2. The process of claim 1, wherein the non-nucleophilic base is selected from N,N-diisopropylethylamine (DIPEA), 8-diazabicycloundec-7-ene (DBU), 1,5-diazabicyclo(4.3.0)non-5-ene (DBN), 2,6-di-tert-butylpyridine, tert-butyl-lithium, tert-butyl-phosphazene, lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), potassium tert-butoxide, potassium bis(trimethylsilyl)amide (KHMDS), lithium tetramethylpiperidide (LiTMP), sodium hydride, potassium hydride, and sodium tert-butoxide.

5. 5. The process of claim 4, wherein the non-nucleophilic base is potassium bis(trimethylsilyl)amide (KHMDS).

6. 2. The process of claim 1, wherein the polar aprotic solvent is selected from acetone, acetonitrile, dichloromethane, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethyl acetate, hexamethylphosphoric triamide (HMPT), pyridine, tetrahydrofuran (THF), and mixtures thereof.

7. 10. The process of claim 1, wherein the hydrolysis of the compound of formula IV comprises reacting the compound of formula IV with a nucleophilic base selected from sodium hydroxide and sodium methoxide.

8. The compound of formula II has formula V: 【Chemistry 5】 (In the formula, R ３ is a halogen) 2. The process of claim 1, wherein the compound is prepared by contacting the compound of formula (I) with azetidine in the presence of a palladium catalyst.

9. R ３ The process of claim 8 , wherein is bromine.

10. 9. The process of claim 8, wherein the molar ratio of azetidine to the compound of formula V is from about 2.0 to about 3.

0.

11. The palladium catalyst is Pd(OAc) ２ 10. The process of claim 8, wherein the palladium compound is selected from the group consisting of palladium(II) pivalate, tetrakis(triphenylphosphine)palladium(0), bis(acetonitrile)palladium(II) dichloride, bis(triphenylphosphine)palladium(II) dichloride, [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, tris(dibenzylideneacetone)dipalladium(0) and palladium(II) chloride.

12. The contact may be with trimethylphosphine, triphenylphosphine, tricyclohexylphosphine, tri(o-tolyl)phosphine, 2-(dicyclohexylphosphino)-2',4',6'-tri-i-propyl-1,1'-biphenyl (XPhos), 2-(dicyclohexylphosphino)-2'-(N,N-dimethylamino)biphenyl, 2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl, 2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl, butyldi-1-adamantylphosphine, 2-(di-t-butylphosphino)biphenyl, 2-(dicyclohexylphosphino)biphenyl, (R)-(-)-1- 9. The process of claim 8, carried out in the presence of a ligand selected from [(S)-2-(diphenylphosphino)ferrocenyl]ethyldicyclohexylphosphine, 1,2-bis(diphenylphosphino)benzene (dppbenzene), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (BINAP), bis(2-diphenylphosphinophenyl)ether (DPEphos), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos), 1,4-bis(diphenylphosphino)butane (dppb), 1,2-bis(diphenylphosphino)ethane (dppe), 1,1′-bis(diphenylphosphino)ferrocene (dppf), 1,3-bis(diphenylphosphino)propane (dppp), and [(t-Bu 3 )PH]BF 4 .

13. 13. The process of claim 12, wherein the ligand is xantphos.

14. The compound of formula V has formula VI 【Chemistry 6】 with a compound of formula VII: 【Chemistry 7】 by contacting a compound of formula VIII: 【Chemistry 8】 and thereafter contacting said compound of formula VIII with a reducing agent to obtain said compound of formula V, wherein R ３ The process of claim 8 , wherein is a halogen.

15. The compound of formula VI can be prepared by reacting 1-(3-hydroxyphenyl)ethan-1-one with a compound of formula IX: 【Chemistry 9】 (In the formula, R ４ is a halogen) in the presence of a non-nucleophilic base in a polar aprotic solvent to form 1-(3-cyclobutoxyphenyl)ethan-1-one, which is then contacted with dimethyl oxalate and a strong base to provide the compound of formula VII.

16. The compound of formula I is prepared by reacting the compound of formula 【Chemistry 10】 2. The process of claim 1, further comprising converting the compound to a Tris salt of

17. 17. A Tris salt prepared by the process of claim 16.

18. formula 【Chemistry 11】 or a salt thereof.

19. formula 【Chemistry 12】 or a salt thereof.

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