Processes of making and crystalline forms of mdm2 inhibitor

Abstract

To provide: processes for making 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid and intermediates; processes for making the intermediates; and crystalline forms of the compound and the intermediates.SOLUTION: The invention provides a crystalline compound of the formula in the figure, pharmaceutical compositions comprising the compound, and the like.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention provides a method for producing 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (hereinafter referred to as "Compound A" in this specification) and an intermediate, and a method for producing the intermediate. The present invention also provides crystal forms of the compound and the intermediate.

Background Art

[0002] p53 is a tumor suppressor and a transcription factor that responds to cellular stress by activating the transcription of a number of genes involved in cell cycle arrest, apoptosis, senescence, and DNA repair. Unlike normal cells in which p53 activation rarely occurs, tumor cells are exposed to constant cellular stress resulting from various insults, including hypoxia and activation of apoptosis-promoting oncogenes. Therefore, there is a strong selective advantage in the inactivation of the p53 pathway in tumors, and it has been proposed that the elimination of p53 function may be a prerequisite for tumor survival. Based on this idea, three groups of researchers used mouse models to demonstrate that the absence of p53 function is a continuous requirement for the maintenance of established tumors. When the researchers restored the p53 function of tumors with inactivated p53, the tumors disappeared.

[0003] p53 is inactivated by mutations and / or deletions in 50% of solid tumors and 10% of liquid tumors. Other major elements of the p53 pathway are also genetically or epigenetically altered in cancer. The oncoprotein MDM2 inhibits p53 function, is activated by gene amplification, and has been reported to have an incidence of up to 10%. In addition, MDM2 is inhibited by another tumor suppressor, p14ARF. Changes downstream of p53 may be at least partly responsible for the inactivation of the p53 WT (p53 wild-type) pathway in tumors. Supporting this idea, some p53 WTThe tumor retains its ability to arrest the cell cycle, but shows a reduced ability to undergo apoptosis. One cancer treatment strategy involves the use of small molecules that bind to MDM2 and abrogate its interaction with p53. MDM2 inhibits p53 activity through three mechanisms: 1) acting as an E3 ubiquitin ligase to promote p53 degradation, 2) binding to the p53 transcriptional activation domain to block it, and 3) transporting p53 from the nucleus to the cytoplasm. All three of these mechanisms are blocked by abrogating the MDM2-p53 interaction. In particular, this treatment strategy is applicable to tumors with WT p53. In tests using small molecule MDM2 inhibitors, promising tumor growth reduction was achieved both in vitro and in vivo. Furthermore, in patients with p53-inactivated tumors, stabilization of wild-type p53 in normal tissues by MDM2 inhibition may enable selective protection of normal tissues from cytotoxic agents.

[0004] The present invention relates to compounds that inhibit the interaction between p53 and MDM2 and can activate p53 downstream effector genes. Thus, the compounds of the present invention will be useful for the treatment of cancer, bacterial infections, viral infections, ulcers, and inflammation. In particular, the compounds of the present invention are useful for the treatment of solid tumors such as tumors of the breast, colon, lung, and prostate, as well as liquid tumors such as lymphomas and leukemias. As used herein, MDM2 means the human MDM2 protein, and p53 means the human p53 protein. Note that human MDM2 is also referred to as HDM2 or hMDM2.

[0005] The following chemical structure [Chemical formula] The compound 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid having [a certain property] is disclosed in PCT International Publication Patent WO2011 / 153,509 (Example 362). This compound, which is an MDM2 inhibitor, is being investigated in human clinical trials for the treatment of various cancers. The present invention provides methods for producing the compound and intermediates, as well as methods for producing the intermediates. It also provides crystalline forms of the compound and the intermediates.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Summary of the Invention

[0007] In Embodiment 1, the present invention provides a crystalline

Chemical Formula

[0008] In Embodiment 2, the present invention provides an anhydrous crystalline

Chemical Formula

[0009] In Embodiment 3, the present invention provides an anhydrous crystalline

Chemical Formula

[0010] In Embodiment 4, the present invention provides the anhydrous crystal according to Claim 3, which has an X-ray diffraction pattern substantially shown in FIG. 1.

Chemical formula

[0011] In Embodiment 5, the present invention provides a pharmaceutical composition containing the crystalline

Chemical formula

[0012] In Embodiment 6, the present invention provides a method for treating bladder cancer, breast cancer, colon cancer, rectal cancer, kidney cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer, squamous cell carcinoma, melanoma, acute lymphoblastic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, Burkitt lymphoma, acute myeloid leukemia, chronic myelogenous leukemia, endometrial cancer, head and neck cancer, glioblastoma, osteosarcoma or rhabdomyosarcoma, which comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition containing the crystalline

Chemical formula

[0013] In Embodiment 7, the present invention provides

Chemical formula

[0014] In Embodiment 8, the present invention provides

Chemical formula

[0015] In Embodiment 9, the present invention provides a crystalline

Chemical formula

[0016] In Embodiment 10, the present invention provides a crystalline

Chemical formula

[0017] In Embodiment 11, the present invention provides the crystalline

Chemical formula

[0018] In Embodiment 12, the present invention provides a compound that is

Chemical formula

[0019] In Embodiment 13, the present invention provides a method for producing

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0020] In Embodiment 14, the present invention provides the method according to Embodiment 13, wherein the dehydration condition is azeotropic distillation with toluene.

[0021] In Embodiment 15, the present invention

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0022] In Embodiment 16, the present invention

Chemical formula

Chemical formula

Chemical formula

[0023] In Embodiment 17, the present invention ​​ [Chemical formula] A method for producing [Chemical formula] with lutidine and [Chemical formula] reacting therewith to [Chemical formula] form and provides a method.

[0024] In Embodiment 18, the present invention [Chemical formula] A method for producing [Chemical formula] reacting [Chemical formula] therewith to [Chemical formula] form, which is [Chemical formula] oxidized to [Chemical formula] and converted to and provides a method.

[0025] In Embodiment 19, the present invention provides the method according to Embodiment 18, wherein the oxidation is achieved using ozone.

[0026] In Embodiment 20, the present invention provides the method according to Embodiment 18, wherein the oxidation is achieved using ozone followed by pinic oxidation.

[0027] In Embodiment 21, the present invention

Chemical formula

Chemical formula

[0028] In Embodiment 22, the present invention is such that the oxidation is achieved using ozone followed by pinic oxidation,

Chemical formula

Chemical formula

[0029] In Embodiment 23, the present invention

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0030] In Embodiment 24, the present invention [Chemical formula] and [Chemical formula] are reacted in the presence of a base to provide the method according to Embodiment 23.

[0031] In Embodiment 25, the present invention provides the method according to Embodiment 24, wherein the base is sodium tert-butoxide.

[0032] In Embodiment 26, the present invention provides the method according to Embodiment 23, wherein the oxidation is achieved using RuCl 3 and NaIO 4 .

[0033] In Embodiment 27, the present invention [Chemical formula] of [Chemical formula] is achieved using methanol and water to provide the method according to Embodiment 23.

[0034] In Embodiment 28, the present invention [Chemical formula] and [Chem.] is reacted in the presence of a base, oxidation is achieved using RuCl 3 and NaIO 4 and is achieved using methanol and water, [Chem.] of [Chem.] conversion to is achieved using methanol and water, A method according to Embodiment 23 is provided.

[0035] In Embodiment 29, the present invention provides a compound that is an ethanol adduct. [Chem.]

[0036] In Embodiment 30, the present invention provides a crystalline [Chem.] ethanol adduct.

[0037] In Embodiment 31, the present invention provides a crystalline [Chem.] ethanol adduct, characterized in that its powder X-ray diffraction pattern includes peaks at diffraction angles 2θ of approximately 10.5, 18.2, 20.3, 21, 21.9, and 24.2.

[0038] In Embodiment 32, the present invention provides the crystalline [Chem.] ​Provide an ethanol adduct.

[0039] In Embodiment 33, the present invention

Chemical formula

[0040] In Embodiment 34, the present invention is crystalline

Chemical formula

[0041] In Embodiment 35, the present invention provides a crystalline

Chemical formula

[0042] In Embodiment 36, the present invention provides the crystalline

Chemical formula

[0043] In Embodiment 37, the present invention

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0044] In Embodiment 38, the present invention provides the method according to Embodiment 37, wherein the oxidizing agent is ozone and the acid is hydrochloric acid.

Brief Description of the Drawings

[0045]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Mode for Carrying Out the Invention

[0046] The present invention provides a method for producing 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (hereinafter referred to as "Compound A") and an intermediate, and a method for producing the intermediate. In addition, crystal forms of the compound and the intermediate are also provided.

[0047] The term "comprising" means non-limiting and includes the indicated components without excluding other elements.

[0048] The term "therapeutically effective amount" means an amount of a compound or a combination of therapeutically effective compounds that improves, alleviates or eliminates one or more symptoms of a particular disease or condition, or prevents or delays the onset of one or more symptoms of a particular disease or condition.

[0049] The terms "patient" and "subject" can be used interchangeably and mean animals such as dogs, cats, cows, horses, sheep and humans. A specific patient is a mammal. The term patient includes males (bucks) and females (does).

[0050] The term "pharmaceutically acceptable" means that the substances described, such as the compounds of the present invention or salts of such compounds, or formulations containing such compounds or specific excipients, are suitable for administration to a patient.

[0051] The terms "treating", "treat" or "treatment" include prevention (e.g., prophylactic treatment) and palliative treatment.

[0052] The term "excipient" means any pharmaceutically acceptable additive, carrier, diluent, adjuvant, or other component other than the active pharmaceutical ingredient (API) that is normally included in a formulation and / or administered to a patient.

[0053] The compounds of the present invention can be administered to a patient in a therapeutically effective amount. The compound can be administered alone or as part of a pharmaceutically acceptable composition or formulation. Additionally, the compound or the composition can be administered all at once, for example by bolus injection, or in multiple doses, for example by a series of tablets, and can be delivered substantially uniformly over a period of time, for example using transdermal delivery. It should be noted that the dosage of the compound can be changed over time.

[0054] Also, the compounds of the present invention or pharmaceutically acceptable salts thereof may be administered in combination with one or more additional pharmaceutically active compounds / agents. It should be noted that the additional pharmaceutically active compounds / agents can be conventional small organic chemical molecules and can be macromolecules such as proteins, antibodies, peptide bodies, DNA, RNA or fragments of these macromolecules.

[0055] If a patient is scheduled to receive, or is receiving, a plurality of pharmaceutically active compounds, the compounds can be administered simultaneously or sequentially. For example, in the case of tablets, the active compounds may be in one tablet or in separate tablets that can be administered simultaneously or sequentially in any order. Further, it should be appreciated that the present compositions may be in different forms. For example, while one or more compounds may be delivered via a tablet, another compound may be administered via injection or orally as a syrup. All combinations, delivery methods and administration orders are contemplated.

[0056] The term "cancer" means a physiological state in mammals characterized by uncontrolled cell growth. General classifications of cancer include carcinomas, lymphomas, sarcomas and blastomas.

[0057] The compounds of the present invention can be used to treat cancer. A method for treating cancer comprises administering to a patient in need thereof a therapeutically effective amount of the present compound or a pharmaceutically acceptable salt thereof.

[0058] The compounds of the present invention can be used to treat tumors. A method for treating tumors comprises administering to a patient in need thereof a therapeutically effective amount of the present compound or a pharmaceutically acceptable salt thereof.

[0059] The present invention also relates to the use of the compounds of the present invention in the manufacture of a medicament for the treatment of conditions such as cancer.

[0060] Cancers that can be treated with the compounds of the present invention include, but are not limited to, carcinomas such as cancers of the bladder, breast, colon, rectum, kidney, liver, lung (small cell lung cancer and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin (including squamous cell carcinoma); lymphohematopoietic tumors (including leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma); hematopoietic tumors of the myeloid cell line (including acute and chronic myelogenous leukemia, myelodysplastic syndrome, and promyelocytic leukemia); mesenchymal-derived tumors (including fibrosarcoma and rhabdomyosarcoma, and other sarcomas such as soft tissue and bone sarcomas); central and peripheral nervous system tumors (including astrocytoma, neuroblastoma, glioma, and schwannoma); and other tumors (including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid carcinoma, and Kaposi sarcoma). Other cancers that can be treated with the compounds of the present invention include endometrial cancer, head and neck cancer, glioblastoma, malignant ascites, and blood cancer.

[0061] Specific cancers that can be treated with the compounds of the present invention include soft tissue sarcoma, bone cancer such as osteosarcoma, breast tumor, bladder cancer, Li-Fraumeni syndrome, brain tumor, rhabdomyosarcoma, adrenocortical carcinoma, colorectal cancer, non-small cell lung cancer, and acute myeloid leukemia (AML).

[0062] In certain embodiments of the present invention related to the treatment of cancer, the cancer is identified as p53 wild-type (p53 WT ). In another specific embodiment, the cancer is identified as p53 WT and CDKN2A variants. In another aspect, the present invention provides a diagnostic method for determining which patients should be administered the compounds of the present invention. For example, a sample of a patient's cancer cells can be collected and analyzed to determine the status of the cancer cells with respect to p53 and / or CDKN2A. In one aspect, patients having a cancer with a mutated p53 are less likely to be selected for treatment than patients having a cancer that is p53 WT . In another aspect, p53 WTand cancer patients who also have a mutant CDNK2A protein are selected over patients who do not have these characteristics. The collection of cancer cells for analysis is well known to those skilled in the art. "p53" WT The term "p53" means a protein encoded by genomic DNA sequence no. NC_000017 version 9 (7512445..7531642) (GenBank), a protein encoded by cDNA sequence no. NM_000546 (GenBank), or a protein having GenBank sequence no. NP_000537.3. The term "CDNK2A variant" means a CDNK2A protein that is not wild type. The term "CDKN2A wild type" means a protein encoded by genomic DNA sequence no. 9:21957751-21984490 (Ensembl ID), a protein encoded by cDNA sequence no. NM_000077 (GenBank) or NM_0581959 (GenBank), or a protein having GenBank sequence no. NP_000068 or NP_478102.

[0063] In another aspect, the present invention relates to the use of a compound of the present invention in combination with one or more pharmaceutical agents that are inhibitors of proteins in the phosphatidylinositol 3-kinase (PI3K) pathway. When the compound of the present invention is combined with an inhibitor of a protein in the PI3K pathway, a synergistic effect including an increase in apoptosis and cell lethality was shown in a cancer cell proliferation assay. Examples of proteins in the PI3K pathway include PI3K, mTOR, and PKB (also known as Akt). There are multiple isoforms of the PI3K protein, including α, β, δ, or γ. The PI3K inhibitor that can be used in combination with the compound of the present invention is contemplated to be selective for one or more isoforms. Selectivity means that the compound inhibits one or more isoforms more strongly than other isoforms. Selectivity is a well-known concept to those skilled in the art and can be measured by well-known in vitro activity or cell-based assays. Preferred selectivities include a selectivity of 2-fold, preferably 10-fold, more preferably more than 100-fold for one or more isoforms compared to other isoforms. In one aspect, the PI3K inhibitor that can be used in combination with the compound of the present invention is a PI3Kα-selective inhibitor. In another aspect, the compound is a PI3Kδ-selective inhibitor.

[0064] Examples of PI3K inhibitors that can be used in combination with one or more compounds of the present invention include those disclosed in PCT International Publication Patent WO2010 / 151791, PCT International Publication Patent WO2010 / 151737, PCT International Publication Patent WO2010 / 151735, PCT International Publication Patent WO2010151740, PCT International Publication Patent WO2008 / 118455, PCT International Publication Patent WO2008 / 118454, PCT International Publication Patent WO2008 / 118468, U.S. Patent Application Publication US20100331293, U.S. Patent Application Publication US20100331306, U.S. Patent Application Publication US20090023761, U.S. Patent Application Publication US20090030002, U.S. Patent Application Publication US20090137581, U.S. Patent Application Publication US2009 / 0054405, U.S. Patent Application Publication U.S.2009 / 0163489, U.S. Patent Application Publication US2010 / 0273764, U.S. Patent Application Publication U.S.2011 / 0092504 or PCT International Publication Patent WO2010 / 108074.

[0065] Compounds that inhibit both PI3K and mTOR (dual inhibitors) are known. In yet another aspect, the present invention provides the use of a dual inhibitor of PI3K and mTOR for use in combination with the compounds of the present invention.

[0066] mTOR is a protein in the PI3K pathway. Another aspect of the present invention is the use of an mTOR inhibitor in combination with the compounds of the present invention. Examples of mTOR inhibitors that can be used in combination with the compounds of the present invention include those disclosed in the documents of PCT International Publication Patent WO2010 / 132598 or PCT International Publication Patent WO2010 / 096314.

[0067] PKB (Akt) is also a protein in the PI3K pathway. Another aspect of the present invention is the use of an mTOR inhibitor in combination with the compounds of the present invention. PKB inhibitors that can be used in combination with the compounds of the present invention include those disclosed in the documents of US Patent No. 7,354,944, US Patent No. 7,700,636, US Patent No. 7,919,514, US Patent No. 7,514,566, US Patent Application Publication US2009 / 0270445A1, US Patent No. 7,919,504, US Patent No. 7,897,619 or PCT International Publication Patent WO2010 / 083246A1.

[0068] The combinations of the present invention may also be used together with radiotherapy, hormone therapy, surgery and immunotherapy. These treatment methods are well known to those skilled in the art.

[0069] Since one aspect of the present invention contemplates the treatment of a disease / condition by a combination of pharmaceutically active compounds that can be administered individually, the present invention further relates to combining individual pharmaceutical compositions in kit form. The kit includes two individual pharmaceutical compositions, the compound of the present invention and a second pharmaceutical composition. The kit includes a container for containing the individual compounds, such as a divided bottle or a divided foil packet. Other examples of containers include syringes, boxes and bags. Usually, the kit includes instructions for use regarding the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when the prescribing physician or veterinarian desires a titration of the individual components of the combination.

[0070] Examples of such kits include so-called blister packs. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (such as tablets, capsules, etc.). A blister pack usually consists of a sheet of relatively rigid material, preferably covered with a foil of a transparent plastic material. During the packaging process, depressions are formed in the plastic foil. These depressions are the size and shape of the tablets or capsules to be packaged. Next, the tablets or capsules are placed in these depressions, and the sheet of relatively rigid material is sealed to the plastic foil on the foil surface opposite to the direction in which the depressions are formed. As a result, the tablets or capsules are sealed in the depressions between the plastic foil and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by applying manual pressure to the depressions to form an opening in the sheet at the depression portion. Thereafter, the tablets or capsules can be removed through this opening.

[0071] It may be desirable to provide a memory aid on the kit, for example, in the form of numbers on the side of a tablet or capsule. These numbers are made to correspond to the regimen days on which the designated tablets or capsules must be taken. Another example of such a memory aid is, for example, a calendar printed on a card such as "First week, Monday, Tuesday... etc., Second week, Monday, Tuesday...". Other variations of the memory aid will be readily apparent. A "daily dose" can be one tablet or capsule or a plurality of pills or capsules to be obtained on a given day. Also, the daily dose of the compound of the present invention can be composed of one tablet or capsule, and the daily dose of a second compound can be composed of a plurality of tablets or capsules, and vice versa. The memory aid should reflect this point and assist in the correct administration of the active substance. In another specific embodiment of the present invention, a dispenser designed to dispense the daily dose one by one in the order of use is provided. Preferably, the dispenser is provided with a memory aid to further facilitate compliance with the regimen of the medication. Examples of such memory aids are mechanical counters that indicate the number of daily doses dispensed. Another example of such a memory aid is, for example, a battery-powered microchip memory linked to a liquid crystal readout or audible reminder signal that reads the date on which the daily dose was last taken and / or reminds of the date on which the next dose should be taken.

[0072] The compounds of the present invention and other pharmaceutically active compounds can be administered to a patient, if necessary, orally, rectally, parenterally (e.g., intravenously, intramuscularly, or subcutaneously), into the large intestine, vaginally, intraperitoneally, intravesically, topically (e.g., as a powder, ointment or drops), or as a buccal or nasal medication. All methods used by those skilled in the art to administer pharmaceutically active agents are contemplated.

[0073] Compositions suitable for parenteral injection may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution in injectable sterile solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, vegetable oils (such as olive oil, etc.) and injectable organic esters such as ethyl oleate. Suitable fluidity can be maintained, for example, by the use of coatings such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0074] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Microbial contamination can be prevented by adding various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, etc. It is also desirable to include isotonic agents such as sugars, sodium chloride, etc. Sustained absorption of injectable pharmaceutical compositions can be achieved by using agents that delay absorption such as aluminum monostearate and gelatin.

[0075] Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or (a) fillers or bulking agents such as starch, lactose, sucrose, mannitol, and silicic acid; (b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) wetting agents such as glycerin; (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (a) dissolution retardants such as paraffin; (f) absorption promoters such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glycerol monostearate; (h) adsorbents such as kaolin and bentonite; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In the case of capsules and tablets, the dosage form may also contain buffering agents. Similar types of solid compositions can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0076] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other techniques well known in the art. They may also contain opacifying agents and may be compositions that release a single active compound or multiple active compounds in a delayed manner at specific sites in the intestinal tract. Examples of encapsulating compositions that can be used are polymeric substances and waxes. The active compound may also, where appropriate, be in the form of microcapsules containing one or more of the above excipients.

[0077] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 - butylene glycol, dimethylformamide, oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame seed oil, glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters or mixtures of these substances.

[0078] In addition to such inert diluents, the compositions may also contain adjuvants such as wetting agents, emulsifying agents, suspending agents, sweetening, flavoring, and perfuming agents. In addition to the active compound, suspensions may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, and sorbitan esters, crystalline cellulose, aluminum metahydroxide, bentonite, agar, and tragacanth, or mixtures of these substances.

[0079] Compositions for rectal administration are preferably suppositories and can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or suppository wax. These substances are solid at normal room temperature but liquid at body temperature and thus dissolve in the rectal or vaginal cavity and release the active ingredient.

[0080] Dosage forms for topical administration of the compounds of the present invention include ointments, powders, sprays and inhalants. The active compound is mixed under aseptic conditions with a physiologically acceptable carrier and any preservatives, buffers or propellants that may be required. Ophthalmic formulations, eye ointments, powders and solutions are also contemplated to be within the scope of the present invention.

[0081] The compounds of the present invention can be administered to patients at dosage levels in the range of about 0.1 to about 3,000 mg per day. For a normal adult human weighing about 70 kg, typically a dosage in the range of about 0.01 to about 100 mg per kilogram body weight is sufficient. The specific dosages and dosage ranges that can be used depend on a number of factors including the requirements of the patient, the condition or severity of the disease being treated, and the pharmacological activity of the compound being administered. Determination of the dosage range and optimal dosage for a particular patient is within the ordinary skill in the art.

[0082] The compounds of the present invention can be administered as pharmaceutically acceptable salts, esters, amides or prodrugs. The term "salt" refers to inorganic and organic salts of the compounds of the present invention. The salts can be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in free base or acid form with a suitable organic or inorganic base or acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate and lauryl sulfate, etc. The salts can include cations based on alkali and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. See, for example, "Pharmaceutical Salts" by S.M. Berge et al., J Pharm Sci, 66:1-19 (1977).

[0083] Examples of pharmaceutically acceptable esters of the compounds of the present invention include C 1 ~C 8 alkyl esters. Acceptable esters also include C 5 ~C 7 cycloalkyl esters and arylalkyl esters such as benzyl. C 1 ~C 4 alkyl esters are generally used. The esters of the compounds of the present invention can be prepared according to methods well known in the art.

[0084] Examples of pharmaceutically acceptable amides of the compounds of the present invention include ammonia, primary C 1 ~C 8Alkylamines and secondary C 1 ~C 8 Amides derived from dialkylamines are included. In the case of secondary amines, the amine may also be in the form of a 5- or 6-membered heterocycloalkyl group containing at least one nitrogen atom. Ammonia, C 1 ~C 3 Primary alkylamines and C 1 ~C 2 Amides derived from dialkyl secondary amines are often used. The amides of the compounds of the present invention can be prepared according to methods well known to those skilled in the art.

[0085] The term "prodrug" means a compound that is converted in vivo to produce a compound of the present invention. This conversion can occur by various mechanisms, such as hydrolysis in the blood. Considerations regarding the use of prodrugs are described in T. Higuchi and W. Stella's "Prodrugs as Novel Delivery Systems", Vol. 14 of the A.C.S. Symposium Series, as well as in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987. By way of example, since the compounds of the present invention contain a carboxylic acid functional group, the prodrug is formed by replacing the hydrogen atom of the carboxylic acid group with a (C 1 ~C 8 alkyl, (C 2 ~C1 2) alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having 3 to 9 carbon atoms, 1-(N-(alkoxycarbonyl)aminomethyl having 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolactone-4-yl, di-N,N-(C 1 ~C 2 ) alkylamino (C 2 ~C 3 ) alkyl (such as β-dimethylaminoethyl), carbamoyl-(C 1 ~C 2 ) alkyl, N,N-di(C 1 ~C 2 ) alkylcarbamoyl-(C 1 ~C 2 ) alkyl and piperidino-, pyrrolidino- or morpholino-(C 2 ~ 3 ) alkyl and the like can be formed by substitution with groups.

[0086] The compounds of the present invention may contain asymmetric or chiral centers and thus may exist as different stereoisomers. All stereoisomers of the present compounds and mixtures thereof (including racemic mixtures) are intended to form part of the present invention. Further, the present invention contemplates all geometric and positional isomers. For example, if the present compound contains a double bond, both cis-type and trans-type (represented by Z and E, respectively), as well as mixtures are contemplated.

[0087] A mixture of stereoisomers, such as a mixture of diastereomers, can be separated into individual stereochemical components by known methods such as chromatography and / or fractional crystallization based on physicochemical differences. Enantiomers can also be separated by reacting a mixture of enantiomers with a suitable optically active compound (e.g., an alcohol) to convert the enantiomer mixture into a mixture of diastereomers, separating the diastereomers, and converting the individual diastereomers into the corresponding pure enantiomers (e.g., by hydrolysis). Also, some compounds may be atropisomers (e.g., substituted biaryls).

[0088] The compounds of the present invention may exist in the unsolvated form or in the solvated form with a pharmaceutically acceptable solvent such as water (hydrate), ethanol, etc. The present invention contemplates and includes both the solvated and unsolvated forms described herein.

[0089] Also, the compounds of the present invention may exist in different tautomeric forms. All tautomers of the compounds of the present invention are contemplated. For example, all tautomers of the tetrazole component are included in the present invention. Also, for example, all keto - enol forms or imine - enamine forms of the present compound are included in the present invention.

[0090] One skilled in the art will recognize that the compound names and structures included herein may be based on a particular tautomer of the compound. Although a name or structure indicating only a particular tautomer may be used, unless otherwise specified, it is intended that all tautomers are included by the present invention.

[0091] Also, the present invention is intended to include compounds synthesized in vitro using laboratory techniques, such as techniques well known to synthetic chemists, or compounds synthesized via in vivo techniques such as metabolism, fermentation, digestion, etc. Also, it is contemplated that the compounds of the present invention may be synthesized using a combination of in vitro and in vivo techniques.

[0092] The present invention also includes isotopically labeled compounds that are identical to those recited herein, except that one or more atoms are replaced by atoms having an atomic weight or mass number different from the atomic weight or mass number normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 16 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F and 36 Cl. In one embodiment, the present invention relates to compounds in which one or more hydrogen atoms are replaced by deuterium ( 2 H) atoms.

[0093] Compounds of the present invention containing the foregoing isotopes and / or other isotopes of other atoms are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, for example, compounds incorporating radioactive isotopes such as 3 H and 14 C, are useful in drug and / or substrate tissue distribution assays. Tritium-labeled isotopes, i.e., 3 H, and carbon-14, i.e., 14 C isotopes, are particularly preferred because of the ease of preparation and detection. Furthermore, substitution with heavier isotopes, such as deuterium, i.e., 2 H, can provide certain therapeutic advantages resulting from greater metabolic stability, such as an increase in in vivo half-life or a decrease in dosage requirements, and may thus be preferred in some situations. The isotopically labeled compounds of the present invention can generally be prepared by replacing an isotopically unlabeled reagent with an easily obtainable isotopically labeled reagent.

[0094] The compounds of the present invention may exist in various solid states, including crystalline and amorphous states. The different crystalline, also called polymorphs, and amorphous states of the compounds are contemplated as part of the invention described herein.

[0095] In synthesizing the compounds of the present invention, it may be desirable to use certain leaving groups. The term "leaving group" ("LG") generally refers to a group displaceable by a nucleophile. Such leaving groups are known in the art. Examples of leaving groups include, but are not limited to, halides (e.g., I, Br, F, Cl), sulfonates (e.g., mesylates, tosylates), sulfides (e.g., SCH 3 ), N-hydroxysuccinimide, N-hydroxybenzotriazole, etc. Examples of nucleophiles include, but are not limited to, amines, thiols, alcohols, Grignard reagents, anionic species (e.g., alkoxides, amides, carbanions), etc.

[0096] All patents, published patent applications and other publications cited herein are hereby incorporated by reference.

[0097] The specific experimental examples presented in this application illustrate specific embodiments of the present invention. These examples are meant to be representative and are not intended to limit the scope of the claims in any way.

[0098] 1 H-NMR spectra were typically recorded on a Bruker Avance III 500 spectrometer system (Bruker, Bilerica, MA) equipped with a Bruker 5 mm PABBI probe with a z-axis gradient at 500.13 MHz. 1 H frequency or a Bruker Avance II or Avance III 400 spectrometer equipped with a Bruker 5 mm PABBO probe with a z-axis gradient was used. 1 H frequencies. Samples were typically collected in 500 μL of DMSO-d6 or CD 3 dissolved in either of them. 1 The 1H chemical shift was referenced to the residual solvent signals from δ 2.50 of DMSO-d 6 and δ 3.30 of CD 3 OD.

[0099] Prominent peaks are presented in the table. This usually includes the number of hydrogens, multiplicity (s: singlet, d: doublet, dd: doublet of doublets, t: triplet, q: quartet, m: multiplet, br s: broad singlet), and coupling constants in Hz. Electron ionization (EI) mass spectra were typically recorded on an Agilent Technologies 6140 Quadrupole LC / MS mass spectrometer (Agilent Technologies, Englewood, CO). Mass spectrometry results are shown as mass / charge ratios and, in some cases, subsequently, the relative abundance of each ion (in parentheses). Starting materials in the examples below are typically available from commercial sources such as Sigma-Aldrich, St. Louis, MO, or via literature procedures.

[0100] Powder X-ray diffraction data (XRPD) were obtained using a PANalytical X’Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with a real-time multiple strip (RTMS) detector. The radiation used was CuKα (1.54 Å), and the voltage and current were set to 45 kV and 40 mA, respectively. Data were collected at room temperature at 2θ from 5 to 45° using a step size of 0.0334°. Samples were prepared on a low-background sample holder and placed on a sample stage that rotated with a rotation time of 2 seconds.

[0101] Alternatively, XRPD data were obtained using a PANalytical X’Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with an RTMS detector. The radiation used was CuKα (1.54 Å), and the voltage and current were set at 45 kV and 40 mA, respectively. The data were collected at room temperature with 2θ ranging from 5 to 40° using a step size of 0.0334° in all cases. The sample was prepared on a low-background sample holder and placed on a sample stage that rotated with a rotation time of 2 seconds.

[0102] Alternatively, XRPD data were obtained using a PANalytical X’Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with an RTMS detector. The radiation used was CuKα (1.54 Å), and the voltage and current were set at 45 kV and 40 mA, respectively. The data were collected at room temperature with 2θ ranging from 5 to 40° using a step size of 0.0167° in all cases. The sample was prepared on a low-background sample holder and placed on a sample stage that rotated with a rotation time of 2 seconds.

[0103] Alternatively, XRPD data were obtained using a PANalytical X’Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with an RTMS detector. The radiation used was CuKα (1.54 Å), and the voltage and current were set at 45 kV and 40 mA, respectively. The data were collected at room temperature with 2θ ranging from 3 to 40° using a step size of 0.008°. The sample was prepared on a low-background sample holder and placed on a sample stage with a rotation time of 2 seconds.

[0104] Alternatively, XRPD patterns were obtained using a Bruker D8 Discover X-ray diffractometer (Bruker, Billerica, MA) equipped with an electric xyz sample stage and a GADDS area detector. The radiation used was CuKα (1.54 Å), and the voltage and current were set to 45 kV and 40 mA, respectively. Solid samples on a flat glass plate were mapped, and for each sample, a 1 mm 2 area was scanned in oscillation mode at 2θ from 5 to 48° for 3 minutes.

[0105] Differential scanning calorimetry (DSC) data were collected using a standard DSC mode (DSC Q200, TA Instruments, New Castle, DE). A heating rate of 10 °C / min was employed over a temperature range of 40 °C to 300 °C. The analysis was carried out under nitrogen, and the samples were placed in standard hermetic aluminum pans. Indium was used as the calibration standard.

[0106] Alternatively, DSC data were collected using a temperature-modulated DSC mode (DSC Q200, TA Instruments, New Castle, DE). After equilibrating the sample at 20 °C for 5 minutes, a heating rate of 3 °C / min was employed with an adjustment of ±0.75 °C / min over a temperature range of 20 °C to 200 °C. The analysis was carried out under nitrogen, and the samples were placed in standard non-corrugated aluminum pans. Indium was used as the calibration standard.

[0107]

Table 1

[0108] Procedures and starting materials for manufacturing an intermediate

Chemical formula

Chemical formula

Chem.

[0109] Step A: 2-(3-Chlorophenyl)-1-(4-chlorophenyl)ethanone

Chem.

[0110] Sodium bis(trimethylsilyl)amide (1 M in tetrahydrofuran, 117 mL) was slowly added over 1 hour to a solution of 2-(3-chlorophenyl)acetic acid (10 g, 58.6 mmol) in tetrahydrofuran (58 mL) at -78 °C. After stirring at -78 °C for 40 minutes, a solution of methyl 4-chlorobenzoate (10 g, 58.6 mmol) in tetrahydrofuran (35 mL) was added over 10 minutes. The reaction mixture was stirred at -78 °C for 3 hours and then warmed to 25 °C. After 2 hours at 25 °C, the reaction was quenched with saturated aqueous ammonium chloride solution and most of the tetrahydrofuran was removed under reduced pressure. The residue was extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with saturated sodium chloride solution, dried over sodium sulfate, filtered, and the filtrate was concentrated. The product was recrystallized from ether / pentane to give the title compound as a white solid.

[0111] Alternative procedure A mixture of chlorobenzene (170 L, 1684 mol), 3-chlorophenylacetic acid (50 Kg, 293 mol) and dimethylformamide (0.7 L, 9 mol) at 0 °C was added with thionyl chloride (39.1 Kg, 329 mol) over 30 minutes. The mixture was warmed to 15 °C and stirred for 6 hours. The mixture was cooled to 0 °C and aluminum chloride (43 Kg, 322 mol) was added over 1.5 hours. The mixture was warmed to 20 °C and stirred for 15 hours. Water (200 L) and ethanol (200 L) were added to the mixture and the biphasic mixture was stirred for 2 hours. The phases were separated and the organic phase was washed twice with an aqueous solution of sodium ethylenediaminetetraacetate (3 wt%, 200 L) and once with water (200 L). Heptane (1600 L) was added to the organic phase over 15 minutes. The suspension was stirred for 30 minutes, cooled to -5 °C and filtered. The filtrate was dried at 40 °C for 20 hours. 2-(3-Chlorophenyl)-1-(4-chlorophenyl)ethanone was isolated in a yield of 83.6% (67.4 Kg). 1 H NMR (500 MHz, DMSO-d 6 , δ ppm): 8.05 (m, 2H), 7.62 (m, 2H), 7.33 (m, 3H), 7.21 (br d, J = 7.3 Hz, 1H), 4.45 (s, 2H). MS (ESI) = 265.1 [M+H] + 。

[0112] Step B: Methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate

Chemical Structure

[0113] Methyl methacrylate (12.65 mL, 119 mmol) was added to a solution of 2-(3-chlorophenyl)-1-(4-chlorophenyl)ethanone (30 g, 113 mmol) in tetrahydrofuran (283 mL). Then, potassium tert-butoxide (1.27 g, 11.3 mmol) was added and the reaction mixture was stirred at room temperature for 2 days. The solvent was removed under vacuum and replaced with 300 mL of ethyl acetate. The organic phase was washed with brine (50 mL), water (3 × 50 mL), and brine (50 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated under vacuum to obtain methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate as a mixture of diastereomers in an approximate ratio of 1:1. 1 H NMR (400 MHz, CDCl 3 , δ ppm): 7.87 (m, 2H), 7.38 (m, 2H), 7.27 - 7.14 (a series of m, 4H), 4.61 (m, 1H), 3.69 (s, 1.5H), 3.60 (s, 1.5 H), 2.45 (m, 1H), 2.34 (m, 1H), 2.10 (ddd, J = 13.9, 9.4, 5.5 Hz, 0.5H), 1.96 (ddd, J = 13.7, 9.0, 4.3 Hz, 0.5H), 1.22 (d, J = 7.0 Hz, 1.5H), 1.16 (d, J = 7.0, 1.5 H). MS (ESI) = 387.0 [M + 23] + 。

[0114] Step C: (3S,5R,6R)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one and (3R,5R,6R)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one

Chemical Structure

[0115] The crude product (mainly, (4R,5R)-isopropyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoate) was dissolved in tetrahydrofuran (450 mL) and methanol (150 mL). Lithium hydroxide (1.4 M, 149 mL, 208 mmol) was added and the solution was stirred at room temperature for 24 h. The mixture was concentrated under vacuum and the residue was redissolved in ethyl acetate. 1N aqueous hydrochloric acid was added with stirring until the pH of the aqueous layer was about 1. The layers were separated, the organic phase was washed with brine, dried over magnesium sulfate, filtered and concentrated. This material was dissolved in 200 mL of anhydrous toluene and treated with pyridinium p-toluenesulfonate (PPTS, 0.784 g, 3.12 mmol). The reaction was heated to reflux under Dean-Stark conditions (about 2 h) until the seco acid was consumed. The reaction was cooled to room temperature and washed with saturated sodium bicarbonate (50 mL) and brine (50 mL). This solution was dried over sodium sulfate, filtered and concentrated. The crude material was purified by flash chromatography on silica gel (120 g column, eluting with 100% dichloromethane). The title compound was obtained as a white solid with an enantiomeric ratio of about 94:6 and a 7:3 mixture of methyl diastereomers. 1 H NMR (400 MHz, CDCl 3 , δ ppm): 7.22 - 6.98 (a series of m, 5H), 6.91 (dt, J = 7.4, 1.2 Hz, 0.3H), 6.81 (m, 2H), 6.73 (dt, J = 7.6, 1.4 Hz, 0.7H), 5.76 (d, J = 4.1 Hz, 0.3 H), 5.69 (d, J = 4.7 Hz, 0.7H), 3.67 (dt, J = 6.6, 4.3 Hz, 0.3H), 3.55 (td, J = 7.8, 4.7 Hz, 0.7 H), 2.96 (quintet of d, J = 13.5, 6.7 Hz, 0.7 H), 2.81 (m, 0.3 H), 2.56 (dt, J = 14.3, 8.0 Hz, 0.7 H), 2.32 (dt, J = 13.69, 7.0 Hz, 0.3 H), 2.06 (ddd, J = 13.7, 8.4, 4.1, 0.3 H), 1.85 (ddd, J = 14.1, 12.5, 7.4, 0.7 H), 1.42 (d, J = 7.0 Hz, 0.9 H), 1.41 (d, J = 6.7 Hz, 2.1H). MS(ESI) = 357.0 [M+23] + . [α] D (22 °C, c = 1.0, CH 2 Cl 2 ) = -31.9°; m.p. 98 - 99 °C。

[0116] Step D (3S,5R,6R)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one

Chem.

[0117] A solution of (3S,5R,6R)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one and (3R,5S,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one (4.5 g, 13.4 mmol) and allyl bromide (3.48 mL, 40.3 mmol) in tetrahydrofuran (22 mL) at -35 °C (acetonitrile / dry ice bath) was treated with a solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 17.45 mL, 17.45 mmol). The reaction was warmed to -5 °C over 1 hour and then quenched with 50% saturated ammonium chloride. The reaction was diluted with 100 mL of ethyl acetate and the layers were separated. The organic phase was washed with brine, dried over magnesium sulfate, filtered, concentrated under vacuum, and allowed to stand under vacuum to give the title compound as a white solid. Chiral SFC (92% CO 2, 8% methanol (20 mM ammonia), 5 mL / min, Phenomenex Lux-2 column (Phenomenex, Torrance, CA), 100 bar (10,000 kPa), 40 °C, 5 min method) was used to confirm that the enantiomeric ratio of the compound was 96:4 (major enantiomer: title compound, retention time = 2.45 min, 96%; minor enantiomer (structure not shown: retention time = 2.12 min, 4%). The title compound was recrystallized by adding it to heptane (4.7 g slurried in 40 mL) under reflux, and 1.5 mL of toluene was added dropwise to solubilize it. This solution was cooled to 0 °C. The white solid was filtered and washed with 20 mL of cold heptane to obtain a white powder. Chiral SFC (92% CO 2 , 8% methanol, Phenomenex Lux-2 column, the same method as above) showed an enantiomeric ratio of 99.2:0.8 (major enantiomer: 2.45 min, 99.2%; minor enantiomer: 2.12 min, 0.8%). 1 H NMR (400 MHz, CDCl 3 , δ ppm): 7.24 (ddd, J = 8.0, 2.0, 1.2 Hz, 1H), 7.20 - 7.15 (series of m, 3H), 6.91 (t, J = 2.0 Hz, 1H), 6.78 (br d, J = 7.6 Hz, 1H), 6.60 (m, 2H), 5.84 (ddt, J = 17.6, 10.2, 7.4 Hz, 1H), 5.70 (d, J = 5.3 Hz, 1H), 5.21 - 5.13 (series of m, 2H), 3.82 (dt, J = 11.7, 4.5 Hz, 1H), 2.62 (ABX J AB = 13.7 Hz, J AX = 7.6 Hz, 1H), 2.53 (ABX, J AB = 13.9 Hz, J BX = 7.2 Hz, 1H). 1.99 (dd, J = 14.1, 11.9 Hz, 1H), 1.92 (ddd, J = 13.9, 3.9, 1.2 Hz, 1H). 13 C NMR (CDCl 3, 100 MHz, δ ppm): 175.9, 140.2, 134.5, 134.3, 134.0, 132.2, 129.8, 128.6, 128.0, 127.9, 127.8, 126.4, 119.9, 83.9, 44.5, 42.4, 40.7, 31.8, 26.1. MS(ESI) = 375.2 [M+H] + . IR = 1730 cm -1 . [α] D (24 °C, c = 1.0, CH 2 Cl 2 ) = -191°. m.p. 111 - 114 °C。

[0118] (3S,5R,6R)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one alternative route for production

Chem.

[0119] Step 1: Isopropyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate

Chem.

[0120] A solution of 2-(3-chlorophenyl)-1-(4-chlorophenyl)ethanone (67.4 Kg, 255 mol) in THF (325 L) was azeotropically dried to a water content of 0.05 wt% by Karl Fischer. Methyl methacrylate (25.8 Kg, 257 mol) was added to the solution and the mixture was heated to 45 °C. A solution of potassium tert-butoxide (20 wt% in THF, 14.3 Kg, 25 mol) was added over 30 minutes and the mixture was stirred for 6 hours. The mixture was cooled to 10 °C and an aqueous solution of citric acid monohydrate (20 wt%, 35 L) was added without delay over 5 minutes. Isopropyl acetate (400 L) and an aqueous sodium chloride solution (20 wt%, 300 L) were added. The mixture was stirred for 15 minutes and the phases were separated. While adding isopropanol (350 L) simultaneously, the organic phase was distilled under reduced pressure to give a solution of methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate in isopropanol (54 wt%, total solution mass 140 kg) while producing a distillate of 560 L. The water content of this solution by Karl Fischer was 0.01 wt%. Additional isopropanol (420 L) and sulfuric acid (53 Kg, 535 mol) were added to the solution. The mixture was heated to reflux and stirred for 12 hours, during which 200 L of the solvent was distilled off and 200 L of fresh isopropanol was added to the mixture. The mixture was cooled to 20 °C and water (180 L) was added over 30 minutes. Isopropyl acetate (270 L) was added and the mixture was stirred for 30 minutes. The phases were separated and the aqueous phase was extracted with isopropyl acetate (100 L). The combined organic phases were washed 4 times with water (200 L). While adding isopropanol (50 L) simultaneously, the organic phase was distilled under reduced pressure to give a solution of isopropyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate in isopropanol (60 wt%, total solution mass 134 kg) while producing a distillate of 500 L. The water content of this solution by Karl Fischer was 0.02 wt%. The title substance was obtained in an overall yield of 81% as an approximately 1:1 mixture of diastereomers. 1 H NMR (400 MHz, CDCl 3 , δ ppm): 7.70 - 7.80 (m, 2H), 7.22 - 7.28 (m, 2H), 7.00 - 7.18 (a series of m, 4H), 4.78 - 4.96 (m, 1H), 4.42 - 4.50 (m, 1H), 2.02 - 2.30 (m, 2H), 1.80 - 1.95 (m, 1H), 0.99 - 1.19 (m, 15H).

[0121] Step 2: (3S,5R,6R)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one

Chemical Structure

[0122] To a degassed solution of isopropyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate in isopropanol (60 wt%, total solution mass 252 Kg, starting material isopropanol ester 151 Kg, 385 mol), degassed isopropanol (900 L) and potassium tert-butoxide (13 Kg, 116 mol) were added. A degassed solution of (S)-RUCY®-XylBINAP (also known as RuCl[(S)-diapena][(S)-xylbinap], 230 g, 0.2 mol, catalyst, Takasago International Corporation, Rockleigh, NJ) in isopropanol (25 L) was prepared separately. The mixture was purged 4 times with hydrogen at 5 bar (500 kPa) and stirred at 20 °C for 5.5 hours. The hydrogen pressurization was stopped and the mixture was degassed with nitrogen. Tetrahydrofuran (460 L) was added to the mixture. A solution of lithium hydroxide (24 Kg, 576 mol) in water (305 L) was added to the reaction mixture over 40 minutes and the resulting mixture was stirred at 20 °C for 24 hours. A solution of concentrated hydrochloric acid (79.3 Kg, 11.4 M, 740 mol) in water (690 L) was added to the mixture over 2 hours. Toluene (580 L) was added, the mixture was stirred for 30 minutes and the phases were separated. The aqueous phase was extracted with toluene (700 L). The combined organic layers were washed with an aqueous sodium chloride solution (25 wt%, 700 Kg). While adding toluene (800 L) simultaneously, the organic phase was distilled at atmospheric pressure and 100 °C, resulting in a distillate volume of 2700 L. After this solvent exchange, less than 0.05 wt% of isopropanol or water (by Karl Fischer) remained in the mixture. Carbonyldiimidazole (59 Kg, 365 mol) was added to the toluene solution over 2 hours and the mixture was stirred at 20 °C for an additional 2 hours. The mixture was cooled to 10 °C and a solution of orthophosphoric acid (72 Kg, 545 mol) in water (400 L) was added over 1 hour while maintaining the temperature of the mixture below 20 °C. The mixture was stirred for 30 minutes, the phases were separated and the organic layer was washed with an aqueous sodium chloride solution (25 wt%, 484 Kg). Toluene (400 L) was distilled at atmospheric pressure and 110 °C.After cooling the solution to 20 °C, tetrahydrofuran (500 L) was added, and the water content measured by Karl Fischer was 0.03 wt%. The resulting solution was cooled to -10 °C, and a solution of allyl bromide (66.8 Kg, 552 mol) in tetrahydrofuran (50 L) was added. A solution of lithium hexamethyldisilazide in toluene (255 Kg, 26 wt%, 492 mol) was added over 6 hours, and the mixture was stirred at -10 °C for 1 hour. The mixture was warmed to 0 °C, and an aqueous solution of orthophosphoric acid (40 wt%, 400 mol) was added over 3 hours. The mixture was warmed to 20 °C. Water (200 L) and dichloromethane (400 L) were added. The mixture was stirred for 15 minutes and the phases were separated. The solution was distilled at atmospheric pressure and 100 °C to give a distillate volume of 1350 L, and the residual toluene in the mixture was measured to be 9.8 wt%. The mixture was cooled to 70 °C. Diisopropyl ether (85 L), water (26 L) and isopropanol (65 L) were added. The mixture was cooled to 35 °C, stirred for 9 hours, cooled to 30 °C and filtered. The filtrate was washed three times with heptane (80 L). The solid was dried at 55 °C for 48 hours to give 90.1 Kg of (3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one in an overall yield of 63%. Chiral HPLC showed an enantiomeric ratio of 99.95:0.05.

[0123] Step E (S)-2-((2R,3R)-2-(3-chlorophenyl)-3-(4-chlorophenyl)-3-hydroxypropyl)-N-((S)-1-hydroxy-3-methylbutan-2-yl)-2-methylpenta-4-enamide

Chemical formula

[0124] (3S,5R,6R)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one (113 g, 300.0 mmol) was combined with (S)-2-amino-3-methylbutan-1-ol (93 g, 900.0 mmol), and the suspension was heated at 100 °C for 5 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (1000 mL), and washed with 1 N hydrochloric acid (2×), water, and brine. The organic layer was dried over magnesium sulfate and concentrated under vacuum to give the title compound as a white solid. This was used in the next step without further purification.

[0125] Step F (3S,5S,6R,8S)-8-Allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium trifluoromethanesulfonate

Chemical formula

[0126] To a solution of (S)-2-((2R,3R)-2-(3-chlorophenyl)-3-(4-chlorophenyl)-3-hydroxypropyl)-N-((S)-1-hydroxy-3-methylbutan-2-yl)-2-methylpent-4-enamide (73.7 g, 154 mmol) and 2,6-dimethylpyridine (78 mL, 678 mmol) in dichloromethane (700 mL) at -50 °C, trifluoromethanesulfonic anhydride (57 mL, 339 mmol) was added dropwise via an addition funnel over 60 minutes. The reaction mixture was stirred at -50 °C for an additional 1 hour and concentrated under vacuum to give the title compound as a reddish solid. This was used in the next step without further purification.

[0127] Step G (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylthio)-3-methylbutan-2-yl)-3-methylpiperidin-2-one

Chemical formula

[0128] (3S,5S,6R,8S)-8-Allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium trifluoromethanesulfonate (736 mg, 1.242 mmol) was weighed into an oven-dried 50 mL round-bottom flask and dissolved in 20 mL of dry toluene. To remove trace amounts of moisture in the solid, the toluene was removed under vacuum. This method was repeated twice, and the resulting residue was dried under high vacuum.

[0129] A solution of sodium isopropyl sulfide was prepared by adding potassium 2-methylpropan-2-olate (3.0 mL, 3.00 mmol, 1 M solution in tetrahydrofuran) to a solution of propan-2-thiol (331 mg, 4.35 mmol) in 8 mL of dimethylformamide prepared under nitrogen and cooled to 0 °C. The sulfide solution was stirred at room temperature for 5 minutes and cooled to 0 °C. Dry (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium trifluoromethanesulfonate (736 mg, 1.242 mmol) was dissolved in dimethylformamide (total 8 mL) and transferred via syringe to the sulfide solution over 5 minutes (total 3 times). After 5 minutes, the ice bath was removed and the pale orange solution was warmed to room temperature.

[0130] After stirring overnight, the mixture was partitioned between ethyl acetate and saturated ammonium chloride solution. The aqueous phase was saturated with sodium chloride and back-extracted 3 times. The combined organic materials were washed twice with saturated sodium bicarbonate and twice with brine, dried over sodium sulfate, filtered, and concentrated under vacuum to give a residue. This was purified by silica gel column chromatography (80 g column, gradient elution with 0% - 50% ethyl acetate in hexane).

[0131]

Chem.

[0132] Step A: (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-hydroxy-3-methylbutan-2-yl)-3-methylpiperidin-2-one

Chem.

[0133] To a solution of (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium trifluoromethanesulfonate (Step F above) dissolved in tetrahydrofuran (500 ml) and water (300 ml), lithium hydroxide hydrate (64.6 g, 1540 mmol) was added portionwise over 5 minutes. The reaction mixture was stirred at room temperature for 1 hour and concentrated under vacuum. The residue was dissolved in ethyl acetate (about 1.3 L), and the layers were separated. The organic layer was washed with 1 N hydrochloric acid (cooled with ice, 300 mL × 2 of hydrochloric acid sufficient to protonate and remove the remaining 2,6-dimethylpyridine), water, and brine. The solvent was removed under vacuum to give a residue, which was purified by silica gel column chromatography (1500 g column, elution with a gradient of 0% to 50% ethyl acetate in hexane). The product was also crystallized from cyclohexane.

[0134] Step B: (3S,5S,6R,8S)-8-Allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium 4-methylbenzenesulfonate

Chem.

[0135] (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-hydroxy-3-methylbutan-2-yl)-3-methylpiperidin-2-one (49.77 g, 98 mmol) was transferred to a 1000 mL flask containing 4-methylbenzenesulfonic acid hydrate (19.27 g, 101 mmol) and a stir bar. The reaction mixture was suspended in toluene (230 mL). A Dean-Stark trap and a reflux condenser were attached to the flask, and the stirred mixture was heated to reflux in a pre-heated bath. After 1 hour, the solvent was carefully removed under vacuum, and the resulting residue was dried further under high vacuum. The title compound was used in the next step without further purification.

[0136] Step C (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one

Chemical formula

[0137] (3S,5S,6R,8S)-8-Allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium 4-methylbenzenesulfonate, dry powdered potassium carbonate (26.9 g, 195 mmol), and propan-2-thiol (14 mL, 150 mmol) were added together with 200 mL of freshly sparged dimethylformamide. The mixture was heated at 50 °C under argon. After about 21 h, a solution of meta-chloroperbenzoic acid (68.2 g, 77% purity by weight, in 100 mL of dimethylformamide) was transferred to an addition funnel and rapidly added to the stirred reaction mixture while immersing the flask in an ice bath. After 5 min, the resulting yellow solution was warmed to room temperature. After 10 min, additional meta-chloroperbenzoic acid (12 g, 77% wt%) was added as a solid and the mixture was stirred at room temperature. When complete, the mixture was poured into ethyl acetate and washed with 1 M sodium hydroxide (500 mL) poured into ice. The aqueous phase was back-extracted three times and further washed with 1 M NaOH (500 mL, also poured into ice). The aqueous layer was washed once with ethyl acetate and the organic materials were combined. Sodium thiosulfate (1 M in water, 250 mL) was added to the organic materials in a large Erlenmeyer flask and the mixture was stirred for 20 min. The organic phase was washed again with sodium thiosulfate (1 M in water, 250 mL) and the mixture was allowed to stand over the weekend. The organic materials were concentrated to about 500 mL and then washed successively with 10% aqueous citric acid, 1 M sodium hydroxide, and brine. The organic materials were dried over sodium sulfate, filtered, and concentrated to give the crude product. The residue was purified by flash column chromatography (1.5 kg silica gel column, gradient elution of 0% - 50% ethyl acetate in hexane) to give the title compound as a white solid.

[0138] Synthesis of Compound A (Synthesis A) 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

Chemical Structure

[0139] To a mixture of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylthio)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (390 mg, 0.752 mmol) in acetonitrile (4.0 mL), carbon tetrachloride (4.0 mL), and water (6.0 mL) were added ruthenium(III) chloride trihydrate (22 mg, 0.084 mmol) and sodium periodate (1.12 g, 5.24 mmol). The resulting dark brown mixture was stirred vigorously overnight at ambient temperature. The mixture was filtered through a pad of diatomaceous earth and washed with ethyl acetate. The filtrate was partitioned between 2 M HCl and ethyl acetate. The aqueous phase was back-extracted twice with ethyl acetate, and the combined organic materials were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo to give a residue. This was purified by flash chromatography (40 g silica gel column, eluting with a gradient of 0% to 15% isopropanol in hexane). Fractions containing the desired product were combined, the solvent was removed, and the residue was redissolved in a small amount of ACN / water, frozen, and lyophilized to give a white powder.

[0140] Subsequently, a mixture of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylthio)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (388 mg, 0.748 mmol), ruthenium(III) chloride trihydrate (19.56 mg, 0.075 mmol), and sodium periodate (1.15 g, 5.38 mmol) in acetonitrile (4 mL), carbon tetrachloride (4.00 mL), and water (4.00 mL) was stirred vigorously at ambient temperature. After 4 hours, the mixture was filtered through a pad of diatomaceous earth, and the filtrate was partitioned between ethyl acetate and 2 M HCl. The aqueous phase was back-extracted twice with ethyl acetate, the combined organic materials were washed with brine, dried over sodium sulfate, filtered, and concentrated under vacuum to give a residue. This residue was purified by flash chromatography (40 g silica gel column, eluting with a gradient of 0% - 15% isopropanol in hexane). The fractions containing the product were concentrated and combined with the solid obtained in the previous experiment. The combined material was dissolved in a small amount of acetonitrile / water, frozen, and lyophilized overnight to give a white solid.

[0141] The XRPD pattern obtained was consistent with an amorphous form (Figure 2).

[0142] Synthesis of Compound A (Synthesis B) 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

Chemical Structure

[0143] To a mixture of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (1.73 g, 3.14 mmol) in acetonitrile (18 mL), carbon tetrachloride (18 mL) and water (27 mL) was added sodium periodate (2.85 g, 13.32 mmol) and ruthenium(III) chloride trihydrate (0.049 g, 0.189 mmol). The mixture was stirred vigorously at room temperature for 25 h. The mixture was diluted with 2 M HCl, filtered through a pad of diatomaceous earth and rinsed with ethyl acetate. The organic layer was separated, washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The material was purified twice by flash chromatography (120 g silica gel, eluting with a gradient of 0% - 20% isopropanol in hexane; 120 g column, eluting with a gradient of 0% - 15% isopropanol in hexane). The most pure fractions were concentrated, set aside, and the pooled fractions were combined and chromatographed again using flash chromatography (220 g silica gel; eluting with a gradient of 0% - 20% isopropanol in hexane for 45 min) to purify once more.

[0144] Subsequently, a mixture of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (4.1 g, 7.45 mmol), ruthenium(III) chloride trihydrate (0.120 g, 0.459 mmol), and sodium periodate (6.73 g, 31.5 mmol) in acetonitrile (40 mL), carbon tetrachloride (40 mL), and water (60 mL) was stirred vigorously at ambient temperature for 23 h. The reaction was diluted by the addition of 2 M aqueous HCl, filtered through a pad of diatomaceous earth, and washed with a large volume of ethyl acetate. Most of the organic material was removed under vacuum. The crude product was extracted into ethyl acetate, washed with brine, dried over sodium sulfate, filtered, concentrated, and the residue was purified by flash chromatography twice (330 g silica gel column, eluting with a gradient of 0% - 20% isopropanol in hexane; 330 g silica gel column, eluting with a gradient of 0% - 20% isopropanol in hexane) to afford an off-white foam. The most pure fractions were concentrated, set aside, and the pooled fractions were chromatographed again using a method of pooling and rechromatographing to further purify this material by three additional flash chromatographies (220 g silica gel; eluting with a 0% - 20% isopropanol gradient in hexane for 45 min).

[0145] The combined fractions obtained from both experiments were further purified by two additional flash chromatographies (220 g silica gel; eluting with a 0% - 20% isopropanol gradient in hexane for 45 min), and again, the pure fractions were set aside.

[0146] All of the pure fractions were combined, concentrated under vacuum, dissolved in a small amount of acetonitrile / water, and lyophilized.

[0147] The XRPD pattern was consistent with an amorphous form (Figure 2).

[0148] Synthesis of Compound A (Synthesis C) 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

Chem.

[0149] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (5.05 g, 9.17 mmol) was weighed into a 500 mL round-bottom flask containing a large stir bar and 2.04 g (2.04 g) of sodium periodate. The mixture was diluted with carbon tetrachloride (52 mL), acetonitrile (52 mL) and water (78 mL). The flask was immersed in a water bath at room temperature and the internal temperature was monitored with a digital thermocouple.

[0150] Hydrated ruthenium chloride (about 50 mg) was added all at once. The internal temperature was raised to 22 °C and then ice was added to the bath to cool the mixture. Additional hydrated ruthenium chloride (25 mg) was added after 3 minutes. After stirring for a total of 30 minutes, three portions of sodium periodate (2.08 g, 2.07 g and 2.08 g) were added slowly at 15-minute intervals. The temperature was maintained below 19 °C and ice was quickly added to the bath if the internal temperature began to rise. The mixture was stirred at ambient temperature overnight. The mixture was filtered through a pad of diatomaceous earth and the filter cake was washed copiously with ethyl acetate. The filtrate was concentrated under vacuum and partitioned between 2 M HCl (100 mL) and ethyl acetate (200 mL).

[0151] The title compound was obtained by two flash column chromatographies (330 g silica gel, then 220 g silica gel, gradient elution with 0% - 20% isopropanol in hexane). A portion of this material was lyophilized from acetonitrile and water. Further, two flash column chromatographies (220 g silica gel column, then 330 g, gradient elution with 0% - 20% isopropanol in hexane) were performed to repurify the fractions of low purity. The purest fractions obtained from both experiments were combined, concentrated under vacuum, and lyophilized from acetonitrile and water to obtain the title compound.

[0152] The XRPD pattern was consistent with an amorphous form (Figure 2).

[0153] By the above three syntheses, amorphous Compound A was obtained. No crystalline form was obtained. An attempt to crystallize the amorphous Compound A produced by the above procedure (Synthesis C) is summarized in Table 1A below.

Table 2

[0154] The amorphous compound produced by the above procedure (Synthesis C) was used for high - throughput (HT) crystal polymorphism screening. The starting material was observed to be amorphous by XRPD. In the form screening, out of 192 conditions tested, only one crystalline sample was observed, which is shown as a representative in Figure 5 (Crystal Form 2 of Compound A). The form identified by the HTS screening does not match the anhydrous crystal of Compound A.

[0155] The compound loading was approximately 8 mg / well. The amorphous Compound A (Synthesis C) was dispensed into each well of a rack for 96 - well glass vials. Then, the solid sample in the vial was transferred to a 96 - well crystallization source plate.

[0156] For library design, the crystallization solvent was dispensed into the source plate (960 μL / vial) (Tables 1 and 2). After solvent addition, the source plate was sonicated for 30 minutes, then heated at 55 °C with stirring for 30 minutes and held at 25 °C without stirring for 30 minutes. While maintaining at 25 °C, the solvent in the source plate was aspirated and filtered through a filter plate. Subsequently, the filtrate was aspirated and dispensed into three crystallization plates (evaporation, precipitation, cooling). After completion of filtration of the 96 wells, stirring of the source plate was continued at 25 °C for 8 hours. The evaporation plate (200 μL of filtrate per well) was exposed to the environment for 24 hours. The sealed precipitation plate (150 μL of filtrate per well, injected into either pre-filled 150 μL of poor solvent: water or heptane (Table 1)) was linearly cooled from 25 °C to 5 °C over 8 hours and held at 5 °C for 8 hours. The sealed cooling plate (300 μL of filtrate per well) was started at 25 °C, cooled to 5 °C over 8 hours, and held at 5 °C for an additional 8 hours. At the end of crystallization, the precipitation and cooling plates were centrifuged at 1500 rpm for 10 minutes at 5 °C, and the supernatant of each well of both plates was aspirated and discarded. Before removing each of the four plates and recovering the crystal samples onto a 96-well glass substrate, wicking paper was dipped into each well to ensure drying.

[0157]

Table 3

[0158] Using a cross-polarized light microscope, birefringence images were acquired for each well of the four 96-well plates. XRPD patterns were obtained with a Bruker D8 Discover X-ray diffractometer equipped with a motorized xyz sample stage and an area detector of a General Area Detector Diffraction System (GADDS). The screening samples on a flat glass plate were mapped, and using CuKα radiation (40 kV, 40 mA) through a graphite monochromator and a 0.5 mm pinhole collimator, in oscillation mode at 1 mm for 3 minutes from 5° to 48° 2θ. 2The sample area was scanned. In addition to plate screening, the starting materials were also analyzed using the said equipment and method.

[0159] Furthermore, HT crystallization experiments using bases as additives were carried out. Stoichiometric amounts of CH 3 OK, CH 3 ONa, Tris, and ammonium hydroxide were added as MeOH solutions, and Ca(OH) 2 , lysine, diethanolamine, and diethylamine were added as aqueous solutions, and prior to solvent partitioning, the solvent was evaporated by blowing a nitrogen stream.

[0160] For library design, the crystallization solvent was dispensed into a source plate (960 μL / well). After solvent addition, the source plate was sonicated for 30 minutes, then heated at 55 °C with stirring for 30 minutes and held at 25 °C without stirring for 30 minutes. While maintaining at 25 °C, the solvent in the source plate was aspirated and filtered into a filter plate. Then, the filtrate was aspirated and dispensed into three crystallization plates (evaporation, precipitation, cooling). After 96-well filtration was completed, the source plate was continuously stirred at 25 °C for 8 hours. The evaporation plate (200 μL of filtrate per well) was exposed to the environment for 24 hours. The sealed precipitation plate (150 μL of filtrate per well, injected into 150 μL of pre-filled poor solvent) was linearly cooled from 25 °C to 5 °C over 8 hours and held at 5 °C for 8 hours. The sealed cooling plate (300 μL of filtrate per well) was started at 25 °C and cubically cooled to 5 °C over 8 hours and further held at 5 °C for 8 hours. At the end of crystallization, the precipitation and cooling plates were centrifuged at 1500 rpm for 10 minutes at 5 °C, and the supernatant of each well of both plates was aspirated and discarded. Prior to removing each of the four plates and recovering the crystal samples onto a 96-well glass substrate, wicking paper was dipped into each well to ensure drying.

[0161] No crystalline salts were obtained in these experiments. Crystalline forms that matched the XRPD pattern in Figure 4 (Crystal Form 1 of Compound A) were obtained for seven samples. All crystalline samples observed in the screening of this section were by the evaporation method. The samples were 3 IPA containing OK, MeCN containing Tris, THF / H 2 O (90 / 10) containing lysine, IPA containing lysine, THF / water (90 / 10) containing diethanolamine, MeCN containing diethanolamine, toluene / MeOH (50 / 50) containing diethanolamine. When evaporated, crystalline samples that matched Crystal Form 1 of Compound A by XRPD were obtained.

[0162]

Table 4

[0163] Crystallization studies Experiment 1 2 - ((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (100 mg) was placed in a 13 mm test tube, and 1 mL of 40% ethanol in water was added at room temperature. Even after reflux heating, the substance did not dissolve. An additional 2 mL of 40% ethanol in water was added, but the substance did not completely dissolve even after reflux. Ethanol was added dropwise until the substance became a solution. The solution was slowly cooled. Before reaching room temperature, the substance became oily.

[0164] Experiment 2 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (100 mg) was placed in a 13 mm test tube, dissolved in 1 mL of ethanol, and heated under reflux. Water was added dropwise until the turbidity formed upon addition disappeared within a few seconds (a total of 1 mL of water was added). The solution was cooled slowly. Before reaching room temperature, it became oily. Additional ethanol (0.2 mL) was added and the mixture was heated under reflux. When slowly cooled to room temperature, the substance became oily. Additional ethanol (0.2 mL) was added and the mixture was heated under reflux. The mixture did not become oily upon cooling to room temperature, nor did crystals form. After being left at room temperature for 1.5 hours, when the solution was placed in a freezer, the substance became oily.

[0165] Experiment 3 2-((3R,5R,6S)-5-(3-Chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (100 mg, white foam) was placed in a 13 mm test tube, and 1 mL of 60% ethanol in water was added at room temperature. The foam either completely dissolved or mostly dissolved and then precipitated as a white solid. This solid was collected by vacuum filtration. Analysis showed that this solid was purer than the starting material. 2-((3R,5R,6S)-5-(3-Chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (100 mg, white foam) was placed in a 13 mm test tube, and 1 mL of 60% ethanol in water was added. During the addition, when the mixture was stirred at room temperature, the substance dissolved in a short time and then precipitated as a white solid. The mixture was heated to reflux to dissolve the substance and then slowly cooled to room temperature. After stirring overnight at room temperature, no crystals were formed. When the solid prepared in the above experiment was added to the solution as a seed crystal, a solid was immediately formed. The crystals were collected by vacuum filtration and washed with a cold solution of 60% ethanol in water to obtain a white crystalline solid. Analysis showed a further improvement in purity, and X-ray diffraction showed that the substance was crystalline. XRPD was consistent with the ethanol adduct of compound A (Figure 6).

[0166] Experiment 4 2-((3R,5R,6S)-5-(3-Chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (100 mg, white foam) was placed in a 13 mm test tube, and 0.75 mL of 60% ethanol in water was added. During the addition, when the mixture was stirred at room temperature, after several minutes, the foam was replaced by a white crystalline solid. The mixture was heated to reflux, slowly cooled to room temperature without stirring. After several days, large crystals were formed. This was collected by vacuum filtration to obtain the title compound as colorless needle crystals. A single crystal X-ray structure was obtained and was consistent with the ethanol adduct of compound A (Figure 6).

[0167] Synthesis of Ethanol Adduct of Compound A 2 - ((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

Chemical Structure

[0168] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (86.8 g, 158 mmol) was dissolved in acetonitrile (300 mL) and ethyl acetate (300 mL) and transferred to a 2 L three-necked Morton flask. Water (450 mL) was added. A thermocouple and a magnetic stirrer were attached to the flask and then immersed in a water bath. Ruthenium(III) chloride hydrate (0.782 g, 3.47 mmol) was added, followed by sodium periodate (33.75 g). The temperature rose from 17 °C to 22 °C. After 35 minutes, a second aliquot of sodium periodate (33.75 g) was added and the temperature rose from 21 °C to 25 °C. After 38 minutes, a third aliquot of sodium periodate (33.75 g) was added and the temperature rose from 22 °C to 28 °C over 12 minutes. Ice was added to the water bath and when the mixture had cooled (about 8 minutes), a third aliquot of sodium periodate (35 g) was added. The temperature rose from 21 °C to 25 °C. After stirring overnight at room temperature, sodium periodate (20 g) was added and after 4 hours, another aliquot of sodium periodate (20 g) was added. After 1 hour, the mixture was stirred at room temperature using an overhead stirrer. The reaction mixture was filtered through a Buchner funnel and the filter cake was rinsed with ethyl acetate. The cake was dried in a vacuum filtration apparatus overnight.

[0169] The substance was added to a large separatory funnel together with water (1 L) and ethyl acetate (500 mL). Brine was added (50 mL). After 5 hours, the phases were separated and the organic phase was washed with 10% sodium bisulfite solution. After standing overnight, the phases were separated and the organic phase was washed with brine (1 L). After 30 minutes, the organic phase was separated, dried over sodium sulfate, filtered, and concentrated under vacuum. The crude substance was purified by flash column chromatography (1.5 kg silica gel column, gradient elution with 0% - 50% isopropanol in hexane) to obtain the title compound as a white foam.

[0170] The obtained 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid was dissolved in ethanol and transferred to a 500 mL pear-shaped flask. The solvent was removed under vacuum to obtain a white solid. A 60% ethanol solution in water (360 mL) was added and the mixture was heated to 90 °C to dissolve all the substances. The solution was cooled slowly and about 5 mg of crystalline product was added as a seed crystal at 50 °C, 45 °C, and 40 °C, but the substances dissolved. When about 5 mg of crystalline product was added as a seed crystal to the solution at 37 °C, the substances did not dissolve. The substance was cooled slowly to room temperature and placed in a freezer overnight. The crystals were collected by vacuum filtration through a Buchner funnel and washed with cold 60% ethanol in water (about 100 mL). The substance was dried by suction of air through the filter bed for 4 hours to obtain a white solid (80.6 g). The substance was placed under vacuum at room temperature for 2 days. Next, the substance was placed on a rotary evaporator at 50 °C, 15 torr (2 kPa) for 4 hours. Then, it was placed under vacuum at 50 °C overnight. NMR analysis showed that 6 wt% of ethanol was present in the sample.

[0171] A small portion (100 mg) of the sample was slurried in water (0.5 mL) overnight. The solid was recovered by vacuum filtration, washed with water to obtain a white solid. NMR analysis showed that 2.9 wt% ethanol was present. The substance was slurried again in water (0.5 mL) overnight, recovered by vacuum filtration to obtain a white solid. NMR analysis showed that 0.5 wt% ethanol was present. X-ray diffraction showed that the substance was amorphous.

[0172] The remainder of the substance was heated overnight at 55 °C under vacuum. After cooling to room temperature, it was slurried in water (250 mL) and mechanically stirred. Aliquots were taken periodically to measure the ethanol content of the solid. After 40 h, additional water (100 mL) was added and the substance was stirred at room temperature for a further 4.5 days. The substance was recovered by vacuum filtration to obtain a white granular solid. This was resuspended in water (350 mL) and mechanically stirred at room temperature for about 8 h. The substance was recovered by vacuum filtration through a Buchner funnel to obtain a white solid. This solid was dried for 6 h by drawing air through the filter bed and then exposed to air overnight in a hood to obtain a white solid containing 3.5 wt% ethanol.

[0173] Manual polymorph screening The sample was prepared according to the following general procedure. Approximately 20 mg of the ethanol adduct of Compound A was weighed and added to a 1-drum vial. 1 mL of the solvent was added to the vial. The sample was slurried. The solvents tested were water / ethanol (80 / 20, v / v), water / ethanol (70 / 30, v / v), water / ethanol (60 / 40, v / v), water / 1-propanol (90 / 10, v / v), water / 1-propanol (80 / 20, v / v), water / 1-propanol (70 / 30, v / v), water / acetonitrile (95 / 5, v / v), water / acetonitrile (90 / 10, v / v), water / acetone (95 / 5, v / v), water / acetone (90 / 10, v / v), heptane, heptane / isopropyl acetate (99 / 1, v / v), cyclohexane, cyclohexane / isopropyl acetate (99 / 1, v / v). Observations were recorded at the start of the experiment, at 3 days, 7 days, 10 days, 13 days and 19 days. The sample was analyzed by XRPD at 7 days and 10 days, 13 days or 19 days. The results are shown in Table 3. The XRPD was consistent with the ethanol adduct of Compound A (Figure 6), the propanol solvate of Compound A (Figure 7), the anhydrous crystal of Compound A (Figure 1) or the amorphous form of Compound A (Figure 2).

[0174]

Table 5

[0175] Synthesis of the ethanol adduct of Compound A 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid

Chem.

[0176] Batch 1: (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (80.6 g, 146 mmol) was dissolved in acetonitrile (280 mL) and ethyl acetate (280 mL) and transferred to a 2 L three-necked Morton flask. Water (418 mL) was added. A thermocouple was attached to the flask and it was immersed in a water bath. Ruthenium(III) chloride hydrate (0.726 g, 3.22 mmol) was added, followed by sodium periodate (31.25 g). The temperature rose from 17 °C to 24 °C and ice was added to the water bath to adjust the temperature. After 15 minutes, a second aliquot of sodium periodate (31.25 g) was added and the temperature rose from 18 °C to 20 °C. After 15 minutes, a third aliquot of sodium periodate (31.25 g) was added and the temperature rose from 18 °C to 25.6 °C. Additional ice was added to the water bath. After 10 minutes, a fourth aliquot of sodium periodate (31.25 g) was added. After stirring for 2 hours, sodium periodate (15 g) was added and after 90 minutes, sodium periodate (6 g) was added again. After 1 hour, the liquid was decanted into a large separatory funnel. The solid material was rinsed with ethyl acetate (1.5 L), added to the separatory funnel and washed with 10% sodium bisulfite (1 L). The organic layer was washed with brine and the phases were allowed to separate overnight. The solid material was slurried again with ethyl acetate (300 mL) and filtered. The filtrate was washed with 10% sodium bisulfite and brine. The combined organic layers were dried over sodium sulfate, filtered and concentrated. The crude material was purified by flash column chromatography (1.5 kg silica gel column, eluting with a gradient of 0% - 50% isopropanol in hexane) to give the title compound.

[0177] Batch 2: (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (90.4 g, 162 mmol) was dissolved in acetonitrile (308 mL) and ethyl acetate (308 mL) and transferred to a 2 L three-necked Morton flask. Water (463 mL) was added. A thermocouple and a mechanical stirrer were attached to the flask. Ruthenium(III) chloride hydrate (0.803 g, 3.56 mmol) was added and the reaction vessel was immersed in a cold water bath. Sodium periodate was added in several portions (first portion: 34.0 g) and the temperature of the reaction mixture was monitored to keep it below 25 °C. Ice was added to the water bath periodically to make temperature adjustment easier.

[0178] After stirring for 12 minutes, the second portion was added (39.7 g), followed by the third portion (36.6 g) after 28 minutes, and the fourth portion (35.6 g) after 13 minutes. The mixture was stirred at room temperature overnight, the fifth portion was added (15 g), and the sixth portion (16.5 g) was added after 25 minutes. After about 15 minutes, the reaction mixture was decanted into a separatory funnel and the residual solid was rinsed with ethyl acetate (2 × 1 L). The organic matter was recovered and washed with 10% sodium bisulfite (1 L). The organic layer was washed with brine (1 L), dried over sodium sulfate, filtered, and concentrated. The crude material was purified by flash column chromatography (1.5 kg silica gel column, elution with a gradient of 0% to 20% isopropanol in hexane) to give the title compound.

[0179] Batch 3: (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (131.8 g, 239 mmol) was dissolved in acetonitrile (402 mL) and ethyl acetate (402 mL) and transferred to a 2 L three-necked Morton flask. Water (603 mL) was added. A thermocouple and a mechanical stirrer were attached to the flask. Ruthenium(III) chloride hydrate (1.079 g, 4.79 mmol) was added and the reaction vessel was immersed in a cold water bath. Sodium periodate was added in several portions (first portion: 59 g) and the temperature of the reaction mixture was monitored to keep it below 25 °C. Ice was added to the water bath periodically to facilitate temperature control.

[0180] After stirring for 45 minutes, the second portion was added (50 g), followed by the third portion after 30 minutes (22 g), the fourth portion after 20 minutes (30 g), the fifth portion after 20 minutes (50 g). After stirring for 2 hours, the sixth portion (20 g) was added, followed by the seventh portion after 20 minutes (10 g) and the eighth portion after 20 minutes (10 g). After 15 minutes, the reaction mixture was decanted into a separatory funnel and the residual solid was rinsed with ethyl acetate (2 × 1 L). The organic matter was recovered and washed with 10% sodium bisulfite (1 L). The organic layer was washed with brine (1 L), dried over sodium sulfate, filtered and concentrated. To remove fine particles, the substance was dissolved in dichloromethane, filtered and concentrated. The crude substance was divided into two portions and each was purified by flash column chromatography (1.5 kg silica gel column, elution with a gradient of 0% to 20% isopropanol in hexane) to give the title compound.

[0181] Batch 4: (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (87.3 g, 159 mmol) was dissolved in acetonitrile (302 mL) and ethyl acetate (302 mL), and transferred to a 2 L three-necked Morton flask. Water (453 mL) was added. A thermocouple and a mechanical stirrer were attached to the flask. Ruthenium(III) chloride hydrate (0.786 g, 3.49 mmol) was added, and the reaction vessel was immersed in a cold water bath. Sodium periodate was added in several portions (first portion: 34.5 g), and the temperature was monitored to keep the reaction mixture below 25 °C. Ice was added to the water bath periodically to facilitate temperature control.

[0182] After stirring for 1 hour, the second portion was added (34.4 g), followed by the third portion (34.5 g) after 30 minutes, and the fourth portion (34.5 g) after another 30 minutes. The maximum temperature was 27 °C. After stirring for 3.5 hours, the fifth portion (20 g) was added, followed by the sixth portion (5 g) after 1 hour. After 15 minutes, the reaction mixture was decanted into a separatory funnel, and the residual solid was rinsed with ethyl acetate (2 × 1 L). The organic matter was recovered and washed with 10% sodium bisulfite (1 L). The organic layer was washed with brine (0.5 L), dried over sodium sulfate, filtered, and concentrated. The crude material was purified by flash column chromatography (Biotage SNAP cart, 1.5 kg silica gel column, elution with a gradient of 0% - 50% isopropanol in hexane) to obtain the title compound. The impure fractions were re-purified by flash column chromatography (1.5 kg silica gel column, elution with a gradient of 0% - 20% isopropanol in hexane) to obtain the title compound.

[0183] Batch 5: The impure fractions from Batches 1 to 4 were re-purified by multiple flash column chromatographies (the amount of silica gel varied from 330 g to 1.5 kg, elution with a gradient of 0% - 20% isopropanol in hexane) to obtain the title compound.

[0184] Final Purification: The substances from batches 1 - 5 were combined with a portion (18 g) of the substance from another synthesis. 2 - ((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (400 g) was dissolved in ethanol and concentrated under vacuum to obtain a white crystalline solid. A 60% ethanol solution in water (1900 mL) was added, and the mixture was heated to 80 °C while rotating on a rotary evaporator at atmospheric pressure. After the substance dissolved, the solution was slowly cooled while the flask was mechanically stirred. After 3 hours, the temperature was cooled to 50 °C, and crystalline 2 - ((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid was added to the substance as seed crystals. The solid completely dissolved. After 30 minutes, when seed crystals were added again to the solution (at 45 °C), the substance began to crystallize slowly. When the mixture cooled to room temperature, it was placed in a freezer overnight. The crystals were collected by vacuum filtration through a Buchner funnel. The filter cake was washed with ice-cold 60% ethanol in water and dried under vacuum in the Buchner funnel to obtain a white solid. NMR analysis showed that 7.8 wt% ethanol was present (1 molar equivalent). Water (deionized and filtered (Milli-Q filtration system, EMD Millipore, Billerica, MA)) was added to the solid, and the mixture was mechanically stirred at room temperature overnight. Aliquots were taken periodically to monitor the ethanol content of the solid. After 3 days, the substance was vacuum filtered through Buchner filtration, washed with water (the above deionized and filtered water), and the filter cake was dried by vacuum drawing for 3 hours. The filter cake was air dried in the funnel for 2 days, then this was transferred to a 2 L flask as a white solid and dried under vacuum overnight. NMR analysis showed that 6.2 wt% ethanol was present.

[0185] The XRPD pattern was consistent with the ethanol adduct of Compound A (Figure 6). 1 1H NMR (500 MHz, DMSO-d 6 , δ ppm): 12.43 (br s, 1H), 7.72 (br, 1H), 7.37 (br, 2H), 7.23 (t, J = 7.8 Hz, 1H), 7.17 (d, J = 8.1 Hz, 1H), 7.02 (t, J = 1.9, 1.9 Hz, 1H), 6.99 (br, 1H), 6.98 (dt, J = 7.7, 1.4, 1.4 Hz, 1H), 5.01 (d, J = 11.2 Hz, 1H), 3.84 (dd, J = 14.0, 10.1 Hz, 1H), 3.59 (ddd, J = 13.7, 11.3, 2.9 Hz, 1H), 3.39 (m, 1H), 3.18 (dd, J = 13.9, 1.3 Hz, 1H), 3.06 (ddd, J = 10.6, 8.1, 1.6 Hz, 1H), 2.95 (d, J = 13.7 Hz, 1H), 2.50 (d, J = 13.8 Hz, 1H), 2.12 (t, J = 13.5 Hz, 1H), 2.10 (m, 1H), 2.03 (dd, J = 13.3, 3.0 Hz, 1H), 1.29 (d, J = 6.8 Hz, 3H), 1.29 (d, J = 6.8 Hz, 3H), 1.23 (s, 3H), 0.55 (d, J = 6.6 Hz, 3H), 0.37 (d, J = 6.9 Hz, 3H); mS (ESI) = 568.2 [M+H] + 。

[0186] Synthetic procedure for preparing 2 - ((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methyl-2-oxopiperidin-3-yl)acetic acid (Compound A)

[0187] Scheme 1 - Step 1

Chemical Structure

[0188] Preparation of propane-2-sulfinic acid: Tetrahydrofuran (20 L) was added to a reaction vessel, and the temperature of the vessel was cooled to -50 °C. Sulfur dioxide (3.5 kg, 54.6 mol) was condensed at -50 °C in the reaction vessel. Isopropylmagnesium chloride (2 M in tetrahydrofuran, 21 L, 42 mol) was added to the solution. The reaction mixture was stirred at -10 °C for 30 minutes, and 2.5 N aqueous hydrochloric acid solution (18.5 L, 46.2 mol) was added. The reaction mixture was warmed to 20 °C, and t-butyl methyl ether (10 L) was added. The phases were separated, and the aqueous phase was extracted twice with t-butyl methyl ether (10 L). The combined organic extracts were washed with an aqueous sodium chloride solution (12 wt%, 20 mL) and concentrated under reduced pressure to obtain the desired sulfinic acid in a yield of 82% (3.7 Kg).

[0189] (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one preparation

[0190] Tetrahydrofuran (3.6 L) was added to a solution of propane-2-sulfinic acid (912 g, 8.4 mol) in toluene (7.5 L). While maintaining the temperature of the mixture below 20 °C, sodium t-butoxide (2 M in tetrahydrofuran, 3.6 L, 7.2 mol) was added. The pH of the mixture was measured and was about 6. When the mixture was distilled at atmospheric pressure, a distillate of 6.6 kg was obtained. While maintaining the temperature of the mixture below 30 °C, (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium naphthalene-1-sulfonate hemitoluene solvate (also referred to herein as "hemitoluene solvate of the oxoiminium salt") (3.62 Kg, 5.2 mol) and toluene (7.8 L) were added. While simultaneously adding dimethylacetamide (10.9 L), the mixture was distilled at atmospheric pressure and a distillate of 7.2 Kg was obtained. The mixture was stirred at about 120 °C for 14 hours and cooled to 25 °C. t-Butyl methyl ether (9.1 L) and water (14.5 L) were added to the mixture and the biphasic mixture was stirred until no solid was visible. The phases were separated. The organic phase was washed with water (7.3 L) and saturated aqueous sodium bicarbonate solution (7.1 L). The organic phase was filtered and distilled under reduced pressure while simultaneously adding acetonitrile (21.3 L), and a distillate of 15 Kg was obtained. Water (2 L) was added and (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (160 g, 0.29 mol) was added to the solution as seed crystals at 25 °C (the seed crystal material was prepared by the same procedure as in a previous small-scale experiment). The mixture was stirred at 25 °C for 25 minutes and cooled to 20 °C over about 45 minutes. A mixture of acetonitrile (3.0 L) and water (7.0 L) was added to the reaction mixture over 1.5 hours. The resulting mixture was stirred for 1 hour and filtered. The product was washed with a mixture of acetonitrile (3.6 L) and water (2.4 L).The product was dried under nitrogen to obtain (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (2.9 Kg) in a yield of 86%.

[0191] Preparation of the ethanol adduct of Compound A A solution of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (2.4 Kg, 4.4 mol) in ethyl acetate (8.4 L), acetonitrile (8.6 L) and water (6.5 L) was added ruthenium chloride hydrate (20.5 g, 0.09 mol). While maintaining the temperature of the mixture between 20 °C and 28 °C, sodium periodate (5.0 kg, 23.2 mol) was added in four equal portions over 1.5 h. The mixture was stirred for 2.5 h and filtered through a layer of diatomaceous earth (3.33 Kg). The resulting diatomaceous earth cake was washed with isopropyl acetate (10.4 L) and water (3 L). The filtrate was phase-separated. The organic phase was washed twice with an aqueous sodium chloride solution (25 wt%, 5.5 L), twice with an aqueous solution of sodium chloride and sodium bisulfite (25 wt% sodium chloride and 20 wt% sodium bisulfite, 7.8 L), and once with an aqueous sodium chloride solution (25 wt%, 6.5 L). While adding isopropyl acetate (12.4 L) simultaneously, the organic phase was distilled under reduced pressure. The batch was filtered. Carbon (680 g) was added and the mixture was stirred for 13 h. The mixture was filtered through a layer of diatomaceous earth (1.5 Kg) and the diatomaceous earth cake was washed with isopropyl acetate (8 L). While adding ethanol (16 L) simultaneously, the solution was distilled under reduced pressure, giving a distillate of 24.5 Kg. Heptane (8.5 L) was added and an ethanol adduct of Compound A was added as seed crystals to the solution (the seed crystal material was prepared by the same procedure as in a small-scale experiment conducted previously) (95 g). The mixture was stirred at 20 °C for 40 min and while adding heptane (8.8 L) simultaneously, it was distilled under reduced pressure, giving a distillate of 10.9 Kg. The mixture was stirred for 12 h and filtered. The product was washed with a mixture of ethanol (0.4 L) and heptane (1.6 L). The product was dried under nitrogen to obtain an ethanol adduct of Compound A (1.99 Kg) in 70% yield.

[0192] Preparation of the anhydrous crystal of Compound A: The ethanol adduct of Compound A (1.0 Kg, 1.62 mol) was dissolved in methanol (8.5 L), and the resulting solution was filtered. This solution was warmed to 35 °C, and water (2.5 L) was added. Anhydrous crystals of Compound A (50 g, 0.074 mol) were added as seed crystals to the solution, and it was cooled to 20 °C over 4 hours (the seed crystal material was prepared by the same procedure as in a previous small-scale experiment). Water (2 L) was added over 30 minutes. The mixture was stirred for 30 minutes and filtered. The product was dried under nitrogen to obtain anhydrous crystals of Compound A (0.86 Kg) in a yield of 93%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.37 (s, 1H), 7.36 (bs, 4H), 7.23 (t, 1H, J = 7.9 Hz), 7.16 (ddd, 1H, J = 7.9, 1.9, 1.0 Hz), 7.02 (t, 1H, J = 1.9 Hz), 6.98 (bd, 1H, J = 7.9 Hz), 5.02 (d, 1H, J = 7.9 Hz), 3.84 (dd, 1H, J = 13.4, 10.2 Hz), 3.58 (ddd, 1H, J = 13.5, 11.3, 3.0 Hz), 3.39 (spt, 1H, J = 6.8 Hz), 3.17 (bd, 1H, J = 13.4 Hz), 3.07 (bt, 1H, J = 8.6 Hz), 2.95 (d, 1H, J = 13.9 Hz), 2.51 (d, 1H, J = 13.9 Hz), 2.13 (bt, 1H, J = 13.5 Hz), 2.11 (spt, 1H, J = 6.8 Hz), 2.04 (dd, 1H, J = 13.5, 3.0 Hz), 1.30 (2x d, 6H, J = 6.8 Hz), 1.24 (s, 3H), 0.56 (d, 3H, J = 6.8 Hz), 0.38 (d, 3H, J = 6.8 Hz); exact mass [C 28 H 36 Cl 2 NO 5 S] + : calculated value = 568.1691, measured value M / Z [M + 1] = 568.1686.

[0193] When using a seed crystal in the procedure described in this application, it should be noted that in order to obtain a seed crystal for large-scale synthesis, the seed crystal can usually be obtained by small-scale synthesis according to the procedure described herein.

[0194] Scheme 2 - Procedure 2

Chemical formula

[0195] Preparation of calcium propane-2-sulfinate dihydrate: Tetrahydrofuran (20 L) was added to a reaction vessel and the temperature of the vessel was cooled to -50 °C. Sulfur dioxide (3.5 kg, 54.6 mol) was condensed at -50 °C in the reaction vessel. Isopropylmagnesium chloride (2 M in tetrahydrofuran, 21 L, 42 mol) was added to the solution. The reaction mixture was stirred at -10 °C for 30 minutes and 2.5 N aqueous hydrochloric acid solution (18.5 l, 46.2 mol) was added. The reaction mixture was warmed to 20 °C and t-butyl methyl ether (10 L) was added. The phases were separated and the aqueous phase was extracted twice with t-butyl methyl ether (10 L). The combined organic extracts were washed with aqueous sodium chloride solution (12 wt%, 20 mL) and concentrated under reduced pressure to obtain the desired propane-2-sulfinic acid in a yield of 82% (3.7 Kg). This propane-2-sulfinic acid was dissolved in ethanol (37 L) and a solution of calcium acetate monohydrate (3.0 Kg, 17.1 mol) in water (7.2 L) was added. The resulting mixture was stirred for 1 hour and filtered. The product was washed with a mixture of ethanol (10.8 L) and water (1.1 L). The product was dried under nitrogen to obtain calcium propane-2-sulfinate dihydrate in a yield of 86% (4.26 Kg). 1 H NMR (400 MHz, DMSO-d6) δ 3.37 (s, 4H), 1.88 (spt, 2H, J = 7.0 Hz), 0.92 (d, 12H, J = 7.0 Hz).

[0196] (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one Preparation: Calcium propane-2-sulfinate dihydrate (2943616) (2.7 Kg, 9.36 mol) and toluene (22 L) were added to a 60 L vessel. The reaction mixture was warmed to 110 °C and distilled under reduced pressure while simultaneously adding toluene (43 L), resulting in a distillate of 50 Kg. The reaction mixture was cooled to 40 °C, and (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium naphthalene-1-sulfonate hemitoluene solvent (3.6 Kg, 5.2 mol) and toluene (9.0 L) were added. The reaction mixture was warmed to 110 °C and distilled at atmospheric pressure while simultaneously adding dimethylacetamide (10.9 L), resulting in a distillate of 15.8 Kg. The mixture was stirred at about 120 °C for 14 hours and cooled to 40 °C. t-Butyl methyl ether (9.1 L) and water (14.5 L) were added to the mixture, and the two-phase mixture was stirred until no solid was visible. The phases were separated. The organic phase was washed twice with water (2 × 7.3 L), once with saturated aqueous sodium bicarbonate solution (7.1 L), and once with aqueous sodium chloride solution (12 wt%, 7.1 L). The organic phase was cooled to 20 °C, filtered, and distilled under reduced pressure while simultaneously adding acetonitrile (21.3 L), resulting in a distillate of 15 Kg. Water (2 L) was added. (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (160 g, 0.29 mol) was added to the solution as a seed crystal at 25 °C. The mixture was stirred at 25 °C for 25 minutes and cooled to 20 °C over about 45 minutes (the seed crystal material was prepared by the same procedure as in a previous small-scale experiment). A mixture of acetonitrile (3.0 L) and water (7.0 L) was added to the reaction mixture over 1.5 hours. The resulting mixture was stirred for 1 hour and filtered. The product was washed with a mixture of acetonitrile (3.6 L) and water (2.4 L).The product was dried under nitrogen, and (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (2.8 Kg) was obtained in a yield of 83%.

[0197] Preparation of the ethanol adduct of Compound A: A solution of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (1.6 Kg, 2.9 mol) in a mixture of water (2.4 L) and acetonitrile (21.6 L) was flowed at a flow rate of 60 mL / min at 20 °C into the ozone vessel (1 L vessel) of a continuous stirred tank reactor (alternatively, ozonolysis was carried out in the reaction vessel using an ozone sparger). The reaction mixture was added to a solution of sodium chlorite (80 wt%, 1.0 Kg, 11.6 mol) in water (5.6 L) over 6 hours (alternatively, an aqueous solution of sodium chlorite was added to the reaction mixture). The reaction mixture was stirred for 16 hours, and a solution of sodium bisulfite (1.2 Kg, 11.6 mol) in water (5.6 L) was added over 2 hours. The mixture was stirred for 1 hour and the phases were separated. Isopropyl acetate (8 L) and water (8 L) were added to the organic phase. The mixture was stirred for 30 minutes and the phases were separated. The organic phase was washed once with an aqueous sodium chloride solution (6 wt%, 8 L), three times with a 1 M aqueous sodium phosphate solution (pH 6, 8 L), and once with an aqueous sodium chloride solution (6 wt%, 8 L). The organic phase was filtered. When the mixture was distilled under reduced pressure while simultaneously adding isopropyl acetate (32 L), a distillate of 35 Kg was obtained. When the mixture was distilled under reduced pressure while simultaneously adding ethanol (32 L), a distillate of 36 Kg was obtained. Heptane was added (9.6 L), and when the mixture was distilled under reduced pressure, a distillate of 5 Kg was obtained. An ethanol adduct of compound A (80 g, 0.13 mol) was added as a seed crystal to the mixture (the seed crystal material was prepared by the same procedure as in a previously conducted small-scale experiment). Heptane (6.4 L) was added over 1 hour, the mixture was stirred for 12 hours, cooled to 15 °C, and filtered. The product was washed with a mixture of ethanol (90 mL) and heptane (4.8 L). The product was dried under nitrogen to obtain an ethanol adduct of compound A (1.33 Kg) in a yield of 81%.

[0198] Preparation of the anhydrous crystal of compound A: The ethanol adduct of Compound A (1.0 Kg, 1.62 mol) was dissolved in methanol (8.5 L), and the resulting solution was filtered. This solution was warmed to 35 °C, and water (2.5 L) was added. Anhydrous crystals of Compound A (50 g, 0.074 mol) were added as seed crystals to the solution, and it was cooled to 20 °C over 4 hours (the seed crystal material was prepared by the same procedure as in a previous small-scale experiment). Water (2 L) was added over 30 minutes. The mixture was stirred for 30 minutes and filtered. The product was dried under nitrogen to obtain anhydrous crystals of Compound A (0.86 Kg) in a yield of 93%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.37 (s, 1H), 7.36 (bs, 4H), 7.23 (t, 1H, J = 7.9 Hz), 7.16 (ddd, 1H, J = 7.9, 1.9, 1.0 Hz), 7.02 (t, 1H, J = 1.9 Hz), 6.98 (bd, 1H, J = 7.9 Hz), 5.02 (d, 1H, J = 7.9 Hz), 3.84 (dd, 1H, J = 13.4, 10.2 Hz), 3.58 (ddd, 1H, J = 13.5, 11.3, 3.0 Hz), 3.39 (spt, 1H, J = 6.8 Hz), 3.17 (bd, 1H, J = 13.4 Hz), 3.07 (bt, 1H, J = 8.6 Hz), 2.95 (d, 1H, J = 13.9 Hz), 2.51 (d, 1H, J = 13.9 Hz), 2.13 (bt, 1H, J = 13.5 Hz), 2.11 (spt, 1H, J = 6.8 Hz), 2.04 (dd, 1H, J = 13.5, 3.0 Hz), 1.30 (2x d, 6H, J = 6.8 Hz), 1.24 (s, 3H), 0.56 (d, 3H, J = 6.8 Hz), 0.38 (d, 3H, J = 6.8 Hz); exact mass [C 28 H 36 Cl 2 NO 5 S] + : Calculated value = 568.1691, measured value M / Z [M+1] = 568.1686. A representative XRPD pattern of the anhydrous crystal of Compound A is shown in Figure 1.

[0199] An alternative route for producing the anhydrous crystal of Compound A is to produce the DABCO salt instead of the ethanol adduct, as shown in Scheme 3.

[0200] Scheme 3 - DABCO Salt Procedure

Chemical Structure

[0201] Preparation of the DABCO Salt of Compound A: To a stirred solution of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (4.0 Kg, 7.27 mol) in a mixture of water (6 L) and acetonitrile (54 L), ozone was supplied at 20 °C over 10 hours using a Hastelloy C22 inner layer sparger. While maintaining the temperature of the mixture below 40 °C, an aqueous solution of sodium chlorite (80 wt%, 2.5 Kg, 29 mol) in water (14 L) was added over 1 hour. While maintaining the temperature of the reaction mixture below 40 °C, the reaction mixture was stirred for 12 hours and a solution of sodium bisulfite (3.0 Kg, 29 mol) in water (14 L) was added over 2 hours. The mixture was stirred for 1 hour and the phases were separated. Isopropyl acetate (IPAC) (20 L) and 1 M aqueous sodium phosphate solution at pH 6 (8 L) were added to the organic phase. The mixture was stirred for 30 minutes and the phases were separated. The organic phase was washed with 1 M aqueous sodium phosphate solution at pH 6 (20 L) and then with 1 M aqueous sodium chloride solution (20 L). While adding isopropyl acetate (80 L) simultaneously, the mixture was distilled under reduced pressure, yielding a distillate of 75 Kg. The water content of the solution by Karl Fischer was less than 1 percent. The organic phase was filtered. The solution was further distilled to a volume of about 16 L. The solution was heated to 55 °C and 1,4-diazabicyclo[2.2.2]octane (DABCO, 424 g, 3.65 mol) was added. A seed crystal of the 1,4-diazabicyclo[2.2.2]octane (DABCO) salt of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (136 g, 0.18 mol) was added as a slurry in isopropyl acetate and heptane (1 / 1, 800 mL). The mixture was stirred at 55 °C for 20 minutes and cooled to 20 °C over 2 hours. Heptane (16.8 L) was added over 1 hour and the mixture was stirred at 20 °C for 12 hours.The product was filtered and the filter cake was washed once with a mixture of isopropyl acetate and heptane (2 / 3, 21 L) and once with a mixture of isopropyl acetate and heptane (1 / 4, 21 L). The product was dried under nitrogen to obtain the DABCO salt of Compound A (4.64 Kg) in an 87% yield (liquid chromatography area percentage (LCAP) 100%, Compound A 78.9 wt%). The DABCO salt of Compound A is a solvate of isopropyl acetate (IPAC) according to Scheme 3. The DABCO salt of Compound A is a purification reference point for increasing the purity of the active pharmaceutical ingredient (Compound A). Usually, when the purity of the crude reaction mixture is 97 - 99 in terms of the liquid chromatography area percentage purity, the crystallization of the DABCO salt can be used to improve the purity to 100 in terms of the liquid chromatography area percentage (no impurities with a liquid chromatography area percentage greater than 0.05). For comparison, when using the ethanol adduct of Compound A as a reference point to increase the purity of the active pharmaceutical ingredient (Compound A), the 97 - 99 liquid chromatography area percentage purity of the crude reaction mixture can be improved to a 99.5 - 99.6 liquid chromatography area percentage purity (in the filtrate, multiple impurities exist with a liquid chromatography area percentage greater than 0.05). 1 H NMR(400MHz,CDCl 3 ):δ ppm 0.49( d,J=6.8 Hz,6H),0.64(d,J=6.4 Hz,6H),1.23(d,J=6.0 Hz,12H),1.41(s,6H),1.43(d,J=7.6 Hz,12H),2.02(s,6H),2.05 - 2.00(m,2H),2.30 - 2.15(m,4H),2.71(d,J=13.2,2H),2.84(dd,J=2.0,13.6,2H),2.90(d,J=13.6 Hz,2H),2.96(s,12H),3.11(sept,J=6.8 Hz,2H),3.67 - 3.22(m,2H),3.55 - 3.48(m,2H),4.07(dd,J=10.4,13.2 Hz, 2H), 4.99 (sept, J = 6.4 Hz, 2H), 5.13 (d, J = 11.2 Hz, 2H), 7.10 - 6.98 (m, 8H), 7.35 - 7.10 (m, 8H), 13.2 (br, 2H). 13 C NMR(101MHz, CDCl 3 ) δ ppm 15.3, 15.7, 20.3, 21.0, 21.4, 21.8, 25.6, 32.6, 39.6, 41.5, 44.5, 44.6, 44.8, 47.0, 54.8, 58.4, 67.6, 69.2, 76.7, 77.0, 77.4, 125.7, 126.9, 128.2, 128.5, 129.8, 133.9, 134.0, 137.5, 143.8, 170.7, 174.6, 176.3. m.p. 103 °C。

[0202] Preparation of anhydrous crystals of Compound A (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one 1,4-diazabicyclo[2.2.2]octane (DABCO) salt (8.28 Kg, 5.79 mol) was added to isopropyl acetate (41.4 L) and water (41.4 L). 4M aqueous hydrochloric acid solution (3 L, 12.1 mol) was added to this mixture, and the biphasic mixture was stirred for 30 minutes. The phases were separated, and the organic phase was washed twice with 1M aqueous sodium phosphate solution (25 L) at pH 6 and once with aqueous sodium chloride solution (7 wt%, 33 L). While adding isopropyl acetate (42 L) simultaneously, the mixture was distilled under reduced pressure, and a distillate of 56 Kg was obtained. When both the isopropyl acetate content and the water content by Karl Fischer were measured, they were less than 1 percent in the solution. The organic phase was filtered. While adding acetic acid (45 L) simultaneously, the organic phase was distilled under reduced pressure, and a distillate of 20 kg was obtained. The solution was heated to 60 °C, and deionized water (29 L) was added over 30 minutes. A seed crystal of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (320 g, 0.56 mol) was added as a slurry in acetic acid and deionized water (3 / 2, 1 L). The mixture was stirred at 60 °C for 3 hours and cooled to 20 °C over 6 hours. The mixture was stirred at 20 °C for 12 hours. Deionized water (7 mL) was added over 1 hour, and the mixture was stirred for an additional 1 hour. The product was filtered, and the filter cake was washed once with a mixture of acetic acid and deionized water (1 / 1, 13 L) and three times with deionized water (3 × 65 L). The product was dried under nitrogen to obtain (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutan-2-yl)-3-methylpiperidin-2-one (6.3 Kg) in a yield of 92% (LCAP 100%, 100.3 wt%, acetic acid 320 ppm, water < 100 ppm).

[0203] The synthesis of compound A is shown in Scheme A. An important intermediate in this synthesis is the compound (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium naphthalene-1-sulfonate (also referred to herein as the "oxoiminium salt" or the "oxazolinium salt"). The TfO - or TsO - salts of (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridin-4-ium naphthalene-1-sulfonate were not isolated because they were difficult to crystallize. Crystallization is useful because it can be used to remove impurities generated in the procedure or present in the starting materials. Therefore, hydrolysis to the crystalline lactam and subsequent reformation of the oxoiminium salt can be used.

[0204] Scheme A [Chemical formula]

[0205] The present invention describes a method for producing an oxoiminium naphthalenesulfonate, particularly a hemitoluene solvate of a crystalline oxoiminium naphthalenesulfonate. The use of the hemitoluene solvate of the oxoiminium naphthalenesulfonate provides an improvement in the method for producing compound A (see Scheme B below).

[0206] Scheme B [Chemical formula]

[0207] (3S,5R,6S)-3-Allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-hydroxy-3-methylbutan-2-yl)-3-methylpiperidin-2-one and 1-naphthalenesulfonic acid in toluene were heated under dehydration conditions to produce the hemitoluene hydrate of the oxoiminium salt. According to NMR, DSC, and XRPD, this crystalline substance exhibits the characteristics of a hemitoluene solvate. This crystal form is a storage-stable substance and is thus well-suited as a reagent for producing compound A. One method for producing the oxoiminium salt is by ion exchange using 1-naphthalenesulfonate followed by crystallization from toluene. The advantages of using 1-naphthalenesulfonate rather than other counterions include rapid crystallization kinetics, predictable crystal behavior and size, low solubility in toluene at room temperature (<10 mg / mL), a high melting point (207 - 209 °C), and most importantly, a high impurity purging ability. Impurities in all methods, including stereoisomers, were typically purged by single crystallization until the liquid chromatography area percentage (LCAP) was less than 0.5 (see Scheme C below).

[0208] Scheme C

Chemical Structure

[0209] The formation of the oxoiminium salt shown in Scheme D below was achieved by a double dehydration cyclization reaction using Tf 2 O at extremely low temperature conditions (Condition a) or Ts 2 O at high temperature (Condition b).

[0210] Scheme D

Chemical Structure

[0211] The advantage of condition a is that the reaction can be carried out in a single step. However, these conditions can have side reactions (such as undesirable eliminations that lead to stilbene-based by-products) and require undesirable cryogenic treatments. The latter (condition b) is a stepwise method, and the formation of intermediates in the pathway to the oxoiminium naphthalenesulfonate is well characterized. Ts 2 O is a milder reagent, so the unwanted dicyclization reaction is significantly reduced, and a higher yield (>75% vs <60% yield) can be obtained. Furthermore, this method is desirable for scale-up under heating conditions.

[0212] Scheme E [Chemical formula] Ts 2 The stepwise conversion of the valinol adduct (denoted as "amide" in Scheme E) to the oxoiminium naphthalenesulfonate under Ts

[0213] The following is an explanation of a method that enables the supply of multiple kilograms of oxoiminium salts. The first step of this method is to react (3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one with L-valinol at high temperature. The low optical purity (80% ee) and overall purity (85%) of the starting lactone are acceptable. The valinol adduct is formed as a mixture of diastereomers, which is carried over to subsequent synthetic steps.

[0214] The reaction of the valinol adduct (amide in Scheme E) with tosyl anhydride in the presence of 2,6-lutidine is essentially instantaneous at 15 - 25 °C, yielding hydroxyoxazoline as a stable intermediate. In the presence of additional tosyl anhydride and 2,6-lutidine, tosyl oxazoline, a second observable reaction intermediate, is formed. Finally, when the reaction mixture is heated at reflux temperature (55 °C for 1 day) for a long time, the reaction proceeds to completion, giving the oxoiminium tosylate.

[0215] The reaction mixture was quenched with sulfuric acid and washed multiple times with a sodium 1-naphthalenesulfonate solution to facilitate counterion exchange. After a distillation step to switch the reaction solvent from dichloromethane to toluene, the oxoiminium salt was crystallized as a rod-shaped hemitoluene solvate.

[0216] In short, the crystalline oxoiminium salt is a separable and stable intermediate and can be used to purge various impurities such as diastereomers and stilbene by crystallization. As a material for producing Compound A, the hemitoluene solvate of the oxoiminium salt has desirable characteristics such as easy separation with high purity of chemical and stereoisomers, bulk properties suitable for standard manufacturing techniques, and storage stability.

[0217] Scheme F

Chemical formula

[0218] Preparation of the hemitoluene solvate of the oxoiminium salt: According to Scheme F, L-valinol (2.6 Kg, 25.2 mol) was dissolved at 50 °C, and (3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one (3.6 Kg, 84.0 wt%, 80.8% ee, 7.9 mol) was added. The mixture was heated to 110 °C and stirred at that temperature for 5 hours. The mixture was cooled to 20 °C and dichloromethane (17.9 L) was added. 1N aqueous hydrochloric acid solution (18.5 L) was added and the biphasic mixture was stirred for 10 minutes. The phases were separated and the organic phase was washed with an aqueous sodium chloride solution (20 wt%, 7 L). While adding dichloromethane (3.3 L) simultaneously, the organic phase was distilled at atmospheric pressure, and a distillate of 13.7 Kg was obtained. The organic phase was added to a solution of p-toluenesulfonic anhydride (5.9 Kg, 18 mol) in dichloromethane (23.0 L) over 10 minutes. While maintaining the temperature of the mixture below 25 °C, 2,6-lutidine (3.56 Kg, 33.2 mol) was added over 1 hour. The mixture was stirred at 20 °C for 40 minutes. When the mixture was distilled at atmospheric pressure and 40 °C, a distillate of 13.0 Kg was obtained. While maintaining the temperature below 20 °C, the mixture was added to 2N aqueous sulfuric acid solution (19.5 Kg) over 15 minutes. The mixture was stirred for 15 minutes and the phases were separated. The organic phase was washed twice with an aqueous sodium 1-naphthalenesulfonate solution (10 wt%, 19.4 Kg) and once with an aqueous sodium bicarbonate solution (5 wt%, 19.5 Kg). Sodium 1-naphthalenesulfonate dihydrate (64 g, 0.26 mol) was added.

[0219] While adding toluene (27.0 L) simultaneously, the organic phase was maintained at a temperature of 50 °C and distilled under reduced pressure, resulting in a distillate amount of 39.9 Kg. To the mixture, a hemitoluene solvate of an oxoiminium salt (40 g, 0.06 mol) was added as a seed crystal and stirred for 20 minutes (the seed crystal material was prepared by the same procedure as the small-scale experiment conducted previously). The mixture was cooled to 20 °C and stirred for 20 hours. The mixture was filtered. The product cake was washed with toluene (7.9 L) and dried under nitrogen to obtain a hemitoluene solvate of an oxoiminium salt (3.7 Kg, 63.6 wt%, 99.7% ee, 99 / 1 DR) in a yield of 76%. 1 H NMR(400MHz,DMSO-d 6 )d 8.03-8.00(m,1H),7.93-7.90(m,3H),7.56-7.42(m,6.5 H),7.33(s,1H),7.27-7.13(m,6H),5.85(m,1H),5.35(m,3H),5.02(m,1H),4.93(t,1H,J=9.98 Hz),4.3(m,1H),4.09(m,1H),2.79(m,2H),2.39(t,1H,J=13.3 Hz),2.3(s,1.5 H),2.01(dd,1H,J=13.69,3.13 Hz),1.34(s,3H),0.61(d,3H,J=6.46 Hz),0.53(d,3H,J=6.85 Hz),0.41(m,1H)

[0220] Anhydrous oxoiminium salt 1 g of a toluene solvate of an oxoiminium salt was dissolved in chloroform (10 mL), and this solution was concentrated under reduced pressure. Chloroform (10 mL) was added to the resulting residue, and the solution was concentrated again under reduced pressure. Finally, chloroform (10 mL) was added to the resulting residue, and the solution was concentrated under reduced pressure. 1 H NMR(400MHz,CDCl 3 )d 9.13 (d, 1H, J = 8.61 Hz), 8.35 (d, 1H, J = 7.24 Hz), 7.86 (t, 2H, J = 9.0 Hz), 7.57 (m, 1H), 7.48 (m, 2H), 7.28 (m, 5H), 7.09 (m, 3H), 6.11 (d, 1H, J = 11.15 Hz), 5.81 (m, 1H), 5.54 (m, 1H), 5.32 (m, 2H), 4.79 (m, 1H), 4.64 (dd, 1H, J = 9.00, 4.89 Hz), 3.56 (m, 1H), 2.89 (t, 1H, J = 13.69 Hz), 2.65 (m, 2H), 1.97 (dd, 1H, J = 14.08, 3.33 Hz), 1.54 (s, 3H), 0.66 (s, 3H), 0.36 (m, 1H), 0.59 (s, 3H)

Claims

1. Below formula: 【Chemical 1】 1. A method for preparing a compound of the formula: 【Chemistry 2】 is reacted with 1,4-diazabicyclo[2.2.2]octane (DABCO) to give the compound of the formula: 【Chemistry 3】 forming a compound of formula (I).

2. Below formula: 【Chemistry 4】 is reacted under oxidizing conditions to form a compound of the formula: 【Chemistry 5】 10. The method of claim 1, further comprising forming a compound of formula:

3. 3. The method of claim 2, wherein the oxidation conditions include reaction with ozone followed by Pinnic oxidation.

4. The formula below: 【Chemistry 6】 1. A method for preparing a compound of the formula: 【Chemistry 7】 with an acid to form a compound of the formula: 【Chemistry 8】 forming a compound of formula (I).

5. 5. The method of claim 4, wherein the acid is hydrochloric acid.

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