Processes for preparation of selective estrogen receptor degraders

Abstract

To provide methods of making selective estrogen receptor degraders (SERDs), as well as intermediates thereof, salts thereof including a pharmaceutically acceptable salt, and pharmaceutical compositions thereof.SOLUTION: Optionally, there are provided a SERD-activating compound represented by the following formula A having the R-enantiomer form, and a method for producing the compound. In the formula, either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H; R7 is H or PG; and PG is an alcohol protecting group.SELECTED DRAWING: None

Description

[Background technology]

[0001] Selective estrogen receptor degraders (SERDs) bind to estrogen receptors (ERs) and downregulate ER-mediated transcriptional activity. The degradation and downregulation caused by SERDs can be useful in the treatment of various proliferative immune-mediated disorders, cell proliferation disorders, including cancers such as breast cancer, ovarian cancer, endometrial cancer, prostate cancer, uterine cancer, gastric cancer, and lung cancer, as well as new resistance mutations. Some examples of small molecules of SERDs have been disclosed in the literature (see, for example, WO2005 / 073204, WO2014 / 205136, and WO2016 / 097071). Nevertheless, new SERDs are needed to treat ER-positive cancers, such as breast cancer, gastric cancer, and / or lung cancer.

[0002] As described in U.S. Pat. No. 10,654,866 (the '866 patent), a series of SERDs of the following formula have been discovered, along with pharmaceutically acceptable salts thereof: [ka] wherein one of R1 and R2 is independently Cl, F, —CF3, or —CH3, and the other is H. These SERDs are effective inhibitors of cell proliferation and generally exhibit superior pharmacokinetic (PK) and pharmacodynamic (PD) properties compared to other known and characterized SERDs.

[0003] The '866 patent describes synthetic methods for preparing these compounds that utilize the following intermediates: [ka]

[0004] where R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is hydrogen. These intermediates are SERD-active intermediates because they bind to the estrogen receptor (ER) and downregulate ER-mediated transcriptional activity. Because these intermediates are SERD-active, their use has required special conditions, such as the use of high-level containment facilities, which are expensive and limited in number.

[0005] Disclosed herein are new methods and intermediates that avoid the use of SERD-active intermediates. Furthermore, the potential for SERD-active contaminants is significantly reduced, and sophisticated containment facilities are not required. These new methods utilize environmentally friendly reagents, allow for optimal impurity control, and provide an efficient, cost-effective, and facile synthesis of SERD-active compounds that form crystalline materials, which allow for easy purification of the SERD-active compounds.

[0006] Also disclosed are novel intermediates useful for preparing SERD-active compounds, such as those of Formula A (below), and pharmaceutically acceptable salts thereof. The methods disclosed herein minimize the presence of SERD-active intermediates and impurities, and therefore also minimize the need to use high potency / high containment manufacturing conditions. This significantly improves the efficiency and safety of the methods for making compounds of Formula A, while reducing manufacturing costs. Summary of the Invention

[0007] Compound of Formula A: [ka] or a pharmaceutically acceptable salt thereof, wherein either R or R is independently Cl, F, —CF, or —CH, and the other is H, and R is H or PG, the process comprising the step of preparing a compound of structure 8: [ka] or a salt thereof, wherein R7 is PG or H, and PG is an alcohol protecting group, In a solvent, an amine of structure 9, [ka] or a salt thereof, and a reducing agent. Suitable reducing agents are disclosed elsewhere in this application.

[0008] Intermediates useful for preparing compounds of formula A are also disclosed herein.

[0009] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, providing a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent. Suitable reducing agents are disclosed elsewhere in this application.

[0010] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, providing a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of formula A has an enantiomeric excess of at least about 92%.

[0011] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or a PG containing a C3-C7 alcohol. Compounds of Formula A can be prepared using the methods disclosed herein.

[0012] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, providing a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of formula A contains a C3-C7 alcohol.

[0013] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, and has a purity of at least 98% area and contains less than 1% area of ​​one or more dihydroquinoline or quinoline-based impurities, as defined below. Compounds of Formula A can be prepared using the methods disclosed herein.

[0014] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, providing a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A has a purity of at least 98% area and contains less than 1% area of ​​one or more dihydroquinoline or quinoline-based impurities, as defined below.

[0015] Compound of Formula B [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, and has a purity of at least 98% area and contains less than 1% area of ​​one or more dihydroquinoline or quinoline-based impurities.

[0016] Also disclosed herein is a process for preparing pyridine borane, which process comprises reacting a pyridinium salt with sodium borohydride in a solvent. [Brief explanation of the drawings]

[0017] [Figure 1] Chromatographic overlay of Preparation 18b, Preparation 19, and Alternate Preparation 19 using Chromatography System 1. [Figure 2] This is an enlarged version of Figure 1. [Figure 3] Chromatographic overlay of Preparation 18b, Preparation 19, and Alternate Preparation 19 using Chromatography System 2. DETAILED DESCRIPTION OF THE INVENTION

[0018] Described herein are several methods for preparing compounds of formula A. These methods vary the order in which molecular groups or moieties are added to or otherwise manipulated on the quinoline core, but each method utilizes the reaction between structures 8 and 9. As described below, compounds of formula A contain chiral centers, and the methods described herein can be useful for preparing enantiomerically enriched materials.

[0019] The compound of formula A contains a chiral center, and the methods described herein can be used to prepare racemic or enantiomerically enriched formula A containing predominantly the R or S enantiomer. The R and S enantiomers encompassed by formula A are shown immediately below. The R-enantiomeric form is exemplified in formula B: [ka] The S-enantiomeric form is illustrated in Formula C. [ka]

[0020] Enantiomerically enriched compounds of Formula A, i.e., compounds of Formula B (enriched in the enantiomer of Formula B over Formula C) and Formula C (enriched in the enantiomer of Formula C over Formula B), can be prepared by chiral synthesis (as described herein), or by resolving racemic Formula A to obtain the enantiomerically enriched material using methods known in the art, such as chiral chromatography, or by converting racemic Formula A to diastereomeric salts, separating the diastereomeric salts, converting the diastereomeric salts to a non-salt form, and isolating the enantiomerically enriched compound. While Formula A can be resolved as described above, any of the racemic intermediates disclosed herein can be resolved using methods known in the art, and the resulting enantiomerically enriched compound can be used to prepare enantiomerically enriched compounds of Formula B or Formula C.

[0021] Compounds of formula A, B, and C that can be made using the methods disclosed herein are disclosed in Table 1 (in these compounds, R7 is H). [ka] [Table 1]

[0022] In one embodiment, the compounds of formula A have the R enantiomeric form, ie, they are of formula B.

[0023] In a particularly preferred embodiment, the compound of formula B is [ka] or a pharmaceutically acceptable salt thereof.

[0024] In one embodiment, the compound is the free amine. In another embodiment, the compound is the tosylate salt. In a further embodiment, the compound is prepared according to any of the processes disclosed herein.

[0025] Also described herein are pharmaceutical compositions comprising the compounds of Formula A, Formula B, and Formula C described herein, or pharmaceutically acceptable salts thereof, in combination with a pharmaceutically acceptable excipient, carrier, or diluent. The pharmaceutical compositions described herein can be prepared using pharmaceutically acceptable additives. As used herein, the term "pharmaceutically acceptable additive" refers to one or more carriers, diluents, and excipients that are compatible with other additives in a composition or formulation and are not harmful to the patient. The compounds of Formula A, B, and C described herein, or pharmaceutically acceptable salts thereof, can be formulated as pharmaceutical compositions administered by various routes, such as orally or intravenously. Bioavailability is often a factor in cancer therapy, and the ability to select administration methods and pharmaceutical compositions to control or optimize the bioavailability of active ingredients is useful. For example, orally bioavailable SERD compositions would be particularly useful. The compounds of Formula A, B, and C described herein, or pharmaceutically acceptable salts thereof, are believed to have oral bioavailability. Examples of pharmaceutical compositions and processes for their preparation can be found in "Remington: The Science and Practice of Pharmacy", L.V. Allen Jr., Editor, 22nd Ed., Mack Publishing Co., 2012. Non-limiting examples of pharmaceutically acceptable carriers, diluents, and excipients include: saline, water, starch, sugars, mannitol, and silica derivatives; binders such as carboxymethylcellulose and other cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; kaolin and bentonite; and polyethyl glycols.

[0026] According to various aspects and embodiments described herein, disclosed herein are new methods for preparing compounds of Formula A, B, and C. Compounds of Formula A, B, and C can be prepared using the reagents and reaction schemes shown in the following schemes, preparations, and examples. Certain synthetic steps for each of the described routes can be combined in different ways, or in conjunction with steps from different procedures, to prepare compounds of Formula A, B, and C described herein, or salts thereof, including pharmaceutically acceptable salts.

[0027] The product may be recovered by conventional methods well known in the art, including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. The reagents and starting materials are readily available to one of ordinary skill in the art, such as from commercial sources, or may be prepared using methods known to one of ordinary skill in the art.

[0028] In some embodiments, the present disclosure provides methods and processes for the synthesis of novel intermediates useful in the synthesis of compounds of Formulas A, B, and C described herein. In such embodiments, the present disclosure provides compositions of matter, including novel intermediate compounds and salts thereof. Additionally, certain intermediates described herein may contain one or more protecting groups. The protecting groups may be variable and may be the same or different from one occurrence to the next, depending on the specific reaction conditions and the particular transformation being performed. Typical protection and deprotection conditions are known to the skilled artisan and are described in the literature (see, for example, "Greene's Protective Groups in Organic Synthesis," Fourth Edition, by Peter G.M.Wuts and Theodora W. Greene, John Wiley and Sons, Inc. 2007).

[0029] Individual isomers, enantiomers, and diastereomers can be separated or resolved by one skilled in the art at any convenient point in the synthesis of the compounds of Formulas A, B, and C described herein by methods such as selective crystallization techniques or chiral chromatography (see, e.g., J. Jacques, et al., "Enantiomers, Racemates, and Resolutions," John Wiley and Sons, Inc., 1981, and E.L. Elel and S.H. Wilen, "Stereochemistry of Organic Compounds," Wiley-Interscience, 1994).

[0030] One of skill in the art can also deuterate or tritiate (at least one hydrogen is replaced by deuterium or tritium) the compounds of Formula A, B, and C, and their pharmaceutically acceptable salts, and such molecules are within the scope of the compounds disclosed herein. Deuterated and tritiated compounds are enriched in deuterium and / or tritium to levels beyond those that may be found in nature.

[0031] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 20% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 20% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 20% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 20% compared to a non-isotopically enriched compound.

[0032] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 30% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 30% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 30% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 30% compared to a non-isotopically enriched compound.

[0033] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 40% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 40% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 40% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 40% compared to a non-isotopically enriched compound.

[0034] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 50% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound, in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 50% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 50% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 50% compared to a non-isotopically enriched compound.

[0035] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 60% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 60% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 60% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 60% compared to a non-isotopically enriched compound.

[0036] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 70% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound, in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 70% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 70% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 70% compared to a non-isotopically enriched compound.

[0037] In deuterated compounds, including intermediates disclosed herein, at least one position is isotopically enriched by at least 80% compared to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound, in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are isotopically enriched by at least 80% compared to a non-isotopically enriched compound. In one embodiment, at least three positions are isotopically enriched by at least 80% compared to a non-isotopically enriched compound. In another embodiment, at least four positions are isotopically enriched by at least 80% compared to a non-isotopically enriched compound.

[0038] In deuterated compounds, including intermediates disclosed herein, at least one position is at least 90% isotopically enriched relative to a non-isotopically enriched compound (an isotopically enriched compound is in contrast to a "non-isotopically enriched" compound, in which the percentages of the various isotopes are substantially the same as those present in nature). In one embodiment, at least two positions are at least 90% isotopically enriched relative to a non-isotopically enriched compound. In one embodiment, at least three positions are at least 90% isotopically enriched relative to a non-isotopically enriched compound. In another embodiment, at least four positions are at least 90% isotopically enriched relative to a non-isotopically enriched compound.

[0039] In one aspect, there is provided a process for preparing a compound of formula A or a salt thereof, comprising: [ka] Disclosed herein is a process wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H; and R7 is H or PG; This process produces a compound of the following structure: [ka] or a salt thereof, in a solvent, [ka] or a salt thereof, and a reducing agent.

[0040] In an embodiment, a process may include preparing a compound of structure 8 or a salt thereof, the process including reacting a compound of the structure: [ka] Each R5 is independently a C1-C6 alkyl, or two R5 groups together are -CH2CH2- or -CH2CH2CH2-; and R7 is PG or H, where PG is an alcohol protecting group under hydrolysis conditions of about pH 6 or less. In some embodiments, the hydrolysis conditions include reaction with acid. In one embodiment, R7 is H. In another embodiment, R7 is a protecting group (PG).

[0041] In some embodiments, the process can include preparing a compound of structure 7 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] The reaction includes reacting under basic conditions effective to deprotonate the hydroxyl group and cyclize the compound. In some embodiments, the basic conditions can include a base selected from CsCO, NaH, sodium tert-butoxide, NaOH, LiOH, KOH, and an alkoxide such as methoxide, ethoxide, sodium tert-pentoxide, or potassium tert-pentoxide, wherein the counterion is derived from a Group I or Group II element or a non-nucleophilic base such as DBU. In one embodiment, the base includes at least one of CsCO, sodium tert-pentoxide, or NaOH.

[0042] In some embodiments, the process comprises preparing a compound of structure 6 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] The method comprises reacting the ketone under conditions effective to reduce the ketone to produce the alcohol 6. During the reduction of the ketone 4, some dihydroquinoline 5 may be formed. The dihydroquinoline may be oxidized back to the quinoline 6 using at least one oxidizing agent.

[0043] [ka] Examples of oxidizing agents include, but are not limited to, MnO, DDQ, and oxygen. In some embodiments, the oxidizing agent is MnO. One skilled in the art will also understand that ketones can be reduced under standard reducing conditions such as NaBH, LiBH, LiAlH, NaBHCN, STAB, or by hydrogenation with hydrogen / palladium or hydride reagents to give racemic alcohols, which can undergo chiral resolution.

[0044] In some embodiments, the process can include preparing a compound of structure 4 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] Intermolecular nucleophilic aromatic substitution (S) of the fluoro group with compounds such as 2,2-diethoxyethanol, 2,2-dimethoxyethanol, 2-hydroxymethyl-1,3-dioxane, or 2-hydroxymethyl-1,3-dioxolane, protected aldehyde-2-carbon fragment alcohols N Ar) under conditions effective to form 4. Acetals such as dimethyl acetal, diethyl acetal, or -CHCH- or -CHCHCH- are non-limiting examples of aldehyde protecting groups that can be used in the processes disclosed herein.

[0045] In some embodiments, the process comprises preparing a compound of structure 3 or a salt thereof, the process comprising combining a compound of the following structure: [ka] It includes a cross-coupling reaction between a compound of the following structure: [ka] R6 can be hydrogen or alkyl, or a structure in which two R6 groups are joined by at least two carbons to form a dioxaborolane or dioxaborinane under conditions including a palladium or nickel catalyst effective to form a compound of structure 3.

[0046] Alternatively, in some embodiments, the process comprises reacting a compound of structure 3 or a salt thereof with a compound of structure 17, [ka] can be prepared by reacting a compound of structure 13 (acid chloride or Weinreb amide) or 13a (aldehyde), [ka] Under standard Grignard or lithiation addition reaction conditions, R3 = Cl or HN(OMe)R4 and R4 = C1-C4 alkyl. When 13a is used, the resulting product is an alcohol, which is then oxidized to a ketone using methods disclosed herein or known in the art.

[0047] The compounds of formulas A, B, and C described herein, their pharmaceutically acceptable salts, and pharmaceutical compositions thereof can be used to treat breast cancer, ovarian cancer, endometrial cancer, prostate cancer, uterine cancer, gastric cancer, or lung cancer. The method of treatment includes administering a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable salt thereof to a patient in need thereof. The compounds described herein and their pharmaceutically acceptable salts can be used in the treatment of breast cancer, ovarian cancer, endometrial cancer, prostate cancer, uterine cancer, gastric cancer, and lung cancer, as well as new resistance mutations. Endometrial cancer includes endometrioid endometrial cancer.

[0048] The compounds of formula A, B, and C, and their pharmaceutically acceptable salts, are used in the manufacture of a medicament for treating breast cancer, ovarian cancer, endometrial cancer, prostate cancer, uterine cancer, gastric cancer, or lung cancer, among other diseases and disorders associated with cell proliferation, particularly unregulated cell proliferation caused by ER transcription.

[0049] These and other aspects and embodiments are provided in greater detail in the description that follows.

[0050] Disclosed herein are novel methods for making tetracyclic compounds and their pharmaceutical salts that act as SERDs. The SERDs prepared by the methods and processes described herein inhibit ER-mediated transcription, which may be useful in treating cancers such as breast cancer, ovarian cancer, endometrial cancer, prostate cancer, uterine cancer, gastric cancer, and lung cancer, as well as new resistance mutations. Endometrial cancer includes endometrioid endometrial cancer. These SERDs can be used as single agents or in combination with other classes of drugs, including selective estrogen receptor modulators (SERMs), aromatase inhibitors, CDK4 inhibitors, CDK6 inhibitors, PI3K inhibitors, and mTOR inhibitors, to treat hormone receptor-positive cancers such as breast cancer, gastric cancer, and / or lung cancer. In one aspect, the disclosure provides a process for preparing a compound of formula A or a salt thereof: [ka] wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H; and R7 is H or PG; This process produces a compound of the following structure: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent. Examples of suitable solvents include polar solvents. Polar solvents include alcohols, ethers, nitriles, and some halogenated hydrocarbons. Examples of suitable alcohol solvents include MeOH, EtOH, isopropyl alcohol, tert-butanol, and tert-amyl alcohol, while suitable ethers include THF, cyclopentyl methyl ether, methyl tert-butyl ether, and 2-MeTHF. ACN is an example of a suitable nitrile solvent. Suitable halogenated solvents include DCM, chloroform, and 1,2-dichloroethane. Other solvents that can be used include DMF, NMP, NBP, DMSO, and DMAC. A combination of two or more solvents can be used.

[0051] Examples of reducing agents include, but are not limited to, STAB, LiBH4, NaBH4, NaBH3CN, or pyridine borane. In some reduction protocols, a base such as TEA or DIPEA is also present. In one embodiment, the reducing agent comprises STAB. In another embodiment, the reducing agent comprises NaBH3CN. In yet another embodiment, the reducing agent comprises pyridine borane.

[0052] In an embodiment, the process may include preparing a compound of structure 8 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] In some embodiments, the hydrolysis conditions include reacting under hydrolysis conditions. In some embodiments, the hydrolysis conditions include using an acid.

[0053] In some embodiments, the process can include preparing a compound of structure 7 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] The method includes reacting under basic conditions. The base deprotonates the hydroxyl group, which then reacts to form a cyclized compound. In some embodiments, the basic conditions include a base selected from Cs2CO3, NaH, sodium tert-butoxide, LiOH, NaOH, KOH, and an alkoxide such as methoxide, ethoxide, sodium tert-pentoxide, or potassium tert-pentoxide, and the counterion is derived from a Group I or II element or an organic base such as DBU. In one embodiment, the base includes at least one of Cs2CO3, sodium tert-pentoxide, or NaOH.

[0054] In some embodiments, the process can include preparing a compound of structure 6 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] The method includes reacting under conditions effective to reduce the ketone to an alcohol. Such conditions include the use of a ketone reducing agent. Examples of ketone reducing agents include, but are not limited to, LiAlH, NaBH, or a borane ligand, where the ligand is THF, MeS, catechol, or N,N-diethylaniline.

[0055] In some embodiments, the ketone is enantioselectively reduced to produce the corresponding chiral non-racemic alcohol of structure 6B or 6C. Examples of chiral reduction protocols and / or chiral reducing agents include, but are not limited to, LiAlH4 coordinated to a chiral ligand such as BINOL, chiral borohydride reducing agents, chiral alkylborohydride reducing agents, Corey-Bakshi-Shibata (CBS) reduction involving a borolidone reagent, chiral ligand coordinated to a metal catalyst (such as hydrogen gas and ruthenium coordinated to BINAP), enzymatic reduction, and the use of rhodium coordinated to PyBOX. In one embodiment, the enantioselective reduction conditions include a CBS reduction involving a borolidone reagent.

[0056] In some embodiments, the process can include preparing a compound of structure 4 or a salt thereof, the process comprising reacting a compound of the following structure: [ka] S-fluoro groups with compounds such as 2,2-diethoxyethanol or 2-hydroxymethyl-1,3-dioxane, protected aldehyde-2-carbon fragment alcohols N Ar under conditions effective to form 4. Alkyl and cyclic acetals, such as dimethyl, diethyl acetal, or -CHCH- or -CHCHCH, are non-limiting examples of aldehyde protecting groups.

[0057] In some embodiments, the process can include preparing a compound of structure 3 or a salt thereof, the process comprising combining a compound of the following structure: [ka] It includes a cross-coupling reaction between a compound of the following structure: [ka] R6 can be hydrogen or alkyl, or a structure in which two R6 groups are joined by at least two carbons to form a dioxaborolane or dioxaborinane. In one embodiment, the cross-coupling reaction uses a catalyst. In one embodiment, the catalyst includes a transition metal catalyst. Transition metal catalysts that can be used include palladium catalysts or nickel catalysts. In some embodiments, when a palladium catalyst is used, the palladium catalyst can include XantPhos Pd G2, cataCXium® A Pd G3, bis(triphenylphosphine)palladium(II) chloride, tris(dibenzylideneacetone)dipalladium(0) with tricyclohexylphosphine, (1,1'-bis(diphenylphosphino)ferrocene)palladium(II) chloride, palladium tetrakistriphenylphosphine, or palladium(II) acetate. Suitable nickel catalysts can be NiCl2(dppp), NiCl2(dppf), or G3DenP-Ni.

[0058] definition As used herein, "ACN" refers to acetonitrile, "(Amphos)2PdCl2" refers to bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II), "BSA" refers to bovine serum albumin, "BINAP" refers to 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, "BINOL" refers to 1,1'-bi-2-naphthol, "Bu" refers to butyl, "besylate" refers to CH5SO3- or benzenesulfonate, and "cataCXium® A Pd "G3" refers to [(di(1-adamantyl)-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate, "DBU" refers to 1,8-diazabicyclo(5.4.0)undec-7-ene, "DCM" refers to dichloromethane or methylene chloride, "DDQ" refers to 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, "DEA" refers to diethylamine, "DIPEA" refers to N,N-diisopropylethylamine, "DMAC" refers to dimethylacetamide, "DMF" refers to N,N-dimethylformamide, "DMSO" refers to dimethylsulfoxide, and "ee" refers to enantiomeric excess, which may be measured using chiral gas chromatography or chiral liquid chromatography. "Erα" refers to estrogen receptor α, "EtOAc" refers to ethyl acetate, "EtOH" refers to ethanol or ethyl alcohol, "Ex" refers to example, "FLU" refers to fluorescent unit, "G3DenP-Ni" refers to dendrimer-stabilized nickel nanoparticles, "h" refers to time, "hERα" refers to human estrogen receptor α, and "IC 50" refers to the concentration of a compound that reduces a given response (ligand binding, enzyme response) by 50%, "IPA" refers to isopropyl alcohol or isopropanol, "Me" refers to methyl, "MeOH" refers to methanol or methyl alcohol, "2-MeTHF" refers to 2-methyltetrahydrofuran, "mesylate" refers to CH3SO3- or methyl sulfonate, "min" refers to minutes, "MTBE" refers to methyl tert-butyl ether, "NiCl2(dppf)" refers to 1,1'-bis(diphenylphosphino)ferrocenedichloronickel(II), "NCS" refers to N-chlorosuccinimide, "NiCl2(dppp)" refers to dichloro[1,3-bis(diphenylphosphino)propane]nickel, "NBP" refers to N-butylpyrrolidinone, "NMP" refers to N-methyl-2-pyrrolidone, "PBS" refers to phosphate buffered saline, "PEPPSI™-IPr" refers to pyridine-enhanced pre-catalyst preparation stabilized and initiated—[1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride; "Prep" refers to preparation; "PyBOX" refers to the pyridine linker of the bis(oxazoline) ligand; "RNase" refers to a group of hydrolytic enzymes that degrade ribonucleic acid (RNA) molecules; and "RT" refers to room temperature. "STAB" refers to sodium triacetoxyborohydride, "TEMPO" refers to (2,2,6,6-tetramethylpiperidin-1-yl)oxyl or (2,2,6,6-tetramethylpiperidin-1-yl)oxidanyl, "THF" refers to tetrahydrofuran, "tosylate" refers to HCC6H4SO2- or -OT, "p-TsOH" refers to 4-methylbenzenesulfonic acid or toluenesulfonic acid, "XantPhos Pd G2" refers to chloro[(4,5-bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,1'-biphenyl)]palladium(II), and "XPhos Pd G2" refers to chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) and "XRPD" refers to X-ray powder diffraction.

[0059] All compound names were generated using or derived from Biovia Draw 2017 R2, ChemDraw version 19.1.

[0060] The following schemes, preparations, and examples further illustrate this invention.

[0061] Scheme 1 [ka] Scheme 1 illustrates the preparation of an intermediate of Structure 1. R7 is PG or H, where PG is an alcohol protecting group. Examples of alcohol protecting groups include, but are not limited to, C1-C4 alkyl, benzyl, benzoyl, C1-C6 alkanoyl, methoxyethoxymethyl ether groups, silyl ether groups (e.g., trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, and tri-isopropylsilyl), tetrahydropyranyl, tetrahydrofuranyl, and ethoxyethyl ether. In one embodiment, PG is methyl. In another embodiment, PG is benzoyl.

[0062] In Scheme 1, Step A, 7-methoxy-4-hydroxyquinoline (compound 10) is treated with a halogenating agent such as NCS and heated to about 40° C. in DMF to give compound 11. In Step B, compound 11 is reacted with P(O)Br3 in a solvent such as toluene with heating to about 100° C. to brominate the quinoline ring at the 4-position and displace the hydroxy group to give compound 12 in Step B. Alternatively, this transformation can be carried out under milder conditions using phosphorus tribromide in a solvent such as DMF at about 40° C. to give compound 12.

[0063] In step C, a Grignard reaction is achieved. This reaction involves the addition of an arylmagnesium halide, a Grignard reagent, to a carbonyl group such as an aldehyde (compound 13a) via an organometallic reaction to yield a benzyl alcohol (compound 14). Compound 14 is then oxidized to compound 1 using an oxidizing agent. Examples of oxidizing agents include KMnO, KCrO, pyridinium chlorochromate, Dess-Martin conditions, Swern conditions, Oppenauer conditions, fetizone conditions, and KBr / TEMPO with sodium hypochlorite as the oxidizing agent. In one embodiment, KBr / TEMPO with sodium hypochlorite is used in a biphasic solvent system containing EtOAc / water. Alternatively, after step D, the aryl methyl ether of step D can be demethylated (if PG is methyl) under a variety of conditions recognized by the skilled artisan, such as treatment with BBr or HBr. For example, compound 1 can be treated slowly with BBr3 in a solvent such as DCM, which can be stirred and quenched under conditions to provide the deprotected product of compound 1 (R7 is H), or it can be reacted with aqueous HBr under conditions (e.g., at reflux) to provide the deprotected product of compound 1 (R7 is H).

[0064] Scheme 2 [ka] Scheme 2 shows an alternative method for coupling 12 and 13 to form compound 1, where PG is defined as above and R3 can be Cl, N(OMe)R4, where R4 = C1-C4 alkyl. In Step A, a ketone (compound 1) is generated by adding a Grignard reagent to an acid chloride (R3 = Cl) or Weinreb amide (R3 = N(OMe)R4) with a solvent such as THF at temperatures below 0 °C, preferably about -15 to -25 °C, to yield compound 1. Additionally, organometallic nucleophiles such as organolithium or organozinc variants of the Grignard reagent shown can be used in place of the Grignard reagent.

[0065] Alternatively, after step A, the aryl methyl ether of step A (when PG is methyl) can be demethylated after step D as described in Scheme 1.

[0066] Scheme 3 [ka] In Scheme 3, Step A, compounds 1 and 2 can be used to carry out a Suzuki reaction under basic conditions. The Suzuki reaction is well known in the art as a palladium(0)- or nickel-catalyzed cross-coupling reaction between a boronic acid, boronic ester, or cyclic boronic ester and an organic halide or pseudohalide, such as triflate (OTf). R6 can be hydrogen or alkyl, or two R6 groups can be joined by at least two carbons to form a dioxaborolane or dioxaborinane. Various conditions can be useful to promote such cross-coupling reactions. Alternative cross-coupling reactions include the Negishi, Hiyama, Kumada, or Stille methods. Suitable palladium reagents can include XantPhos Pd G2, cataCXium® A Pd G3 bis(triphenylphosphine)palladium(II) chloride, tris(dibenzylideneacetone)dipalladium(0) with tricyclohexylphosphine, (1,1'-bis(diphenylphosphino)ferrocene)palladium(II) chloride, palladium tetrakistriphenylphosphine, or palladium(II) acetate. Suitable nickel catalysts can be NiCl2(dppp), NiCl2(dppf), or G3DenP-Ni. This reaction can be an efficient, high-yield reaction at low catalyst loadings (e.g., 1 mol% or less). For example, a 4-halogen-substituted quinoline ketone, compound 1 (wherein the halogen is typically Cl, but can also be Br or F), is coupled with an appropriately substituted boronic acid or ester (compound 2), such as 2-fluoro-3 and / or 4-substituted phenylboronic acid, utilizing a catalyst such as Pd-Xphos-G2 described above to form compound 3 and a base such as NaHCO3, K2CO3, or Cs2CO3 in a solvent such as THF and water with heating to about 65°C. Other suitable solvents can be toluene, dioxane, or DMF, and temperatures can range from about room temperature to about 100°C. In some embodiments, a Pd-XPhos-G2 Buchwald-type catalyst can be used in the reaction.Because the reaction uses low palladium levels, palladium scavenging may be minimized or not necessary; however, if desired, Pd scavenging can be performed using silica thiol, which is a silica-bound equivalent of 1-propanethiol.

[0067] In step B, compound 3 is converted to S using 2,2-diethoxyethanol in a polar aprotic solvent and base. N The aldehyde group can be protected using various acetals, such as dimethyl, diethyl acetal, -CH2CH2-, or -CH2CH2CH2-. Other possible polar aprotic solvents include DMF, acetone, ACN, DCM, and EtOAc. This reaction can have a high yield and produce a crystalline product. For example, an electron-deficient aromatic ring compound (compound 3) can be reacted with an electron-rich species, such as 2,2-diethoxyethanol, to form compound 4.

[0068] Those skilled in the art will appreciate that the intermolecular S N It will be appreciated that the Ar reaction may be completed prior to the Suzuki reaction of Step A. Such a rearrangement may require less catalyst and boronic acid / ester, resulting in a more economical process. See Scheme 4 below.

[0069] In Step C, the ketone moiety of compound 4 can be reduced to an alcohol in a chiral or achiral manner. Scheme 4 below describes the chiral reduction of a ketone. The ketone can be reduced under standard reduction conditions. Suitable achiral reducing agents include, but are not limited to, NaBH, LiBH, LiAlH, NaBHCN, or STAB. The resulting racemic alcohol can be subjected to chiral separation and isolation of the desired enantiomer.

[0070] Reduction of the ketone moiety of compound 4 yields a mixture of compounds 5 and 6. Without wishing to be bound by theory, it is believed that the over-reduction of compound 6 to compound 5 is facilitated by the presence of residual Pd from step A. The Pd content of compound 3 and / or compound 4 can be reduced using silica thiol or a similar reagent. In some embodiments, quenching the reduction reaction can produce hydrogen gas, which may require an absorbent. Preferably, the mixture of compounds 5 and 6 is 1 Contains less than about 20%, or less than about 15%, or less than about 10%, or less than about 9%, or less than about 8%, of compound 5 as determined by H NMR.

[0071] The mixture of compounds 5 and 6 is typically not isolated. Instead, it is subjected to oxidation conditions to convert compound 5 to compound 6. This oxidation can be accomplished using a variety of oxidizing agents. Examples of oxidizing agents include oxygen in an inert carrier (to prevent fire) and MnO2. MnO2 is inexpensive and abundant, but is dense and requires good agitation to maintain in suspension. Various forms and particle sizes of MnO2 are available. A packed column (external to the reactor) can be used to avoid suspension and reactor fouling issues. In some embodiments, solvent exchange can be avoided by utilizing ACN as the solvent. In another embodiment, compound 5 can be oxidized to compound 6 using approximately 5% oxygen in nitrogen.

[0072] Following the oxidation reaction in Step D, crude compound 6 is cyclized in Step D1 to form compound 7. Cyclization conditions include treating compound 6 with a suitable base, such as, for example, Cs2CO3, NaH, sodium tert-butoxide, LiOH, NaOH, KOH, an alkoxide base, e.g., sodium, potassium, or lithium methoxide, sodium potassium, or lithium ethoxide, sodium tert-pentoxide, or potassium tert-pentoxide, a non-nucleophilic base, e.g., DBU, or a mixture of two or more thereof. In one embodiment, the base comprises at least one of Cs2CO3, sodium tert-pentoxide, or NaOH. In another embodiment, the base comprises Cs2CO3 or sodium tert-pentoxide. In a further embodiment, the base is Cs2CO3, and the cyclization reaction is carried out at a temperature of about 80-90°C or about 85°C. In another embodiment, the base is sodium tert-pentoxide. In one embodiment, the base is sodium tert-pentoxide and the cyclization reaction is carried out at a temperature of about 20-30°C or about 25°C.

[0073] In some embodiments, undesirable solvents such as dioxane can be avoided by using solvents such as ACN, 2-MeTHF, and / or tert-amyl alcohol. While these three solvents are acceptable, the use of tert-amyl alcohol and / or 2-MeTHF is preferred. In one embodiment, the solvent comprises 2-MeTHF. In another embodiment, the solvent consists of 2-MeTHF. Additionally, the solvent comprises or consists of tert-amyl alcohol.

[0074] In step E, compound 7 can undergo acetal hydrolysis to form aldehyde compound 8. This reaction can be carried out using an acid such as HCl, H2SO4, p-TsOH, methanesulfonic acid, trifluoromethanesulfonic acid, acetic acid, or trichloroacetic acid. In a preferred embodiment, the solvent for hydrolysis includes water and, optionally, a second solvent such as ACN. In another embodiment, p-TsOH is the preferred acid. In a preferred embodiment, at least one of the following hydrolysis systems is used: H2SO4 in aqueous acetone, aqueous HCl in ACN, aqueous HCl in THF, TsOH in water and acetonitrile, or TsOH in aqueous THF can also be used. For example, compound 7 can be deprotected using aqueous HCl in ACN, and the resulting aldehyde 8 forms a precipitate. Alternatively, compound 7 can be deprotected using TsOH in water and acetonitrile. In some embodiments, the pH of the reaction mixture is controlled to about 6 to obtain the aldehyde along with a portion of the hemiacetal. In some embodiments, the aldehyde is isolated as a hydrate, such as a monohydrate or dihydrate. Typically, the hydrate contains about 8% or less water, more typically less than 6%, and even more preferably less than 4%, as determined using Karl Fischer titration. In further embodiments, the aldehyde contains about 3% water, as determined by Karl Fischer titration. The presence of a monohydrate may be due to: 1 Confirmed by 1 H NMR.

[0075] Aldehyde monohydrates often contain some hemiacetal. For example, when R5 is methyl or ethyl, a methyl or ethyl acetal is also formed, respectively. The exact pH of the deprotection reaction will affect the amount of hydrate (mono- and / or di-) and / or acetal present. In some embodiments, bisulfite adducts of aldehydes are prepared. The hydrated aldehydes and hemiacetal forms of aldehydes are converted to aldehydes upon dissolution, e.g., for the reductive amination reaction of Step F.

[0076] In step F, aldehyde 8 is subjected to reductive amination in a solvent using a reducing agent. Examples of suitable solvents include polar solvents. Polar solvents include alcohols, ethers, nitriles, and some halogenated hydrocarbons. Examples of suitable alcohol solvents include MeOH, EtOH, and isopropyl alcohol. Suitable ethers include THF, cyclopentyl methyl ether, methyl tert-butyl ether, and 2-methyl THF (2-MeTHF). ACN is an example of a suitable nitrile solvent. Suitable halogenated solvents include DCM, chloroform, and 1,2-dichloroethane. Other solvents that can be used include dimethylformamide (DMF), NMP, NBP, DMSO, and DMAC. Combinations of two or more solvents can be used. In one embodiment, the alcohol solvent contains isopropanol, tert-butyl alcohol, and / or tert-amyl alcohol. In another embodiment, the alcohol solvent contains isopropanol. In one embodiment, the alcohol solvent contains tert-butyl alcohol. In yet another embodiment, the alcohol solvent contains tert-amyl alcohol. In another embodiment, the solvent comprises THF or 2-MeTHF. In some embodiments, the solvent is THF. In another embodiment, the THF is anhydrous. In other embodiments, the solvent comprises 2-MeTHF. In another embodiment, the 2-MeTHF is anhydrous.

[0077] Reducing agents that can be used include, but are not limited to, LiBH4, NaBH4, NaBH3CN, pyridine borane, STAB, or other boranes. In one embodiment, the reducing agent is STAB. In an alternative embodiment, the reducing agent is pyridine borane. In another embodiment, the reducing agent is NaBH3CN. In some embodiments, the reducing agent is STAB and the solvent comprises THF and / or 2-MeTHF. In some other embodiments, the reducing agent is pyridine borane and the solvent comprises an alcohol.

[0078] In one embodiment, azetidine 9 is a free base. When a salt of azetidine 9 is used, suitable salts include, but are not limited to, an HCl salt, a tosylate salt, a mesylate salt, or a besylate salt. In one embodiment, the azetidine salt is an HCl salt. In another embodiment, the azetidine salt is a tosylate salt. When a salt of azetidine 9 is used, an external base can be added to the reductive amination reaction mixture to at least partially convert the azetidine salt to a free base. A variety of bases can be used, including organic and inorganic bases. Examples of organic bases include, but are not limited to, TEA, DIPEA, and pyridine.

[0079] In one embodiment, the alcoholic solvent comprises EtOH, azetidine salt 9 is used, the reducing agent is STAB, and the external base is TEA or DIPEA. In another embodiment, the alcoholic solvent contains isopropanol, tert-butanol, and / or tert-amyl alcohol, and the reducing agent contains pyridine borane. In another embodiment, the alcoholic solvent contains isopropanol, tert-butanol, and / or tert-amyl alcohol, and the reducing agent contains NaBHCN. In one embodiment, the alcoholic solvent contains tert-amyl alcohol, and the reducing agent contains pyridine borane. In another embodiment, the solvent contains tert-amyl alcohol, the reducing agent contains pyridine borane, and the HCl salt of azetidine 9 is used. In a different embodiment, the solvent contains tert-butyl alcohol, the reducing agent contains pyridine borane, and the HCl salt of azetidine 9 is used. In yet another embodiment, the solvent contains isopropanol, the reducing agent contains pyridine borane, and the HCl salt of azetidine 9 is used. In another embodiment, the solvent contains tert-amyl alcohol, the reducing agent contains pyridine borane, and the tosylate salt of azetidine 9 is used. In a different embodiment, the solvent contains tert-butyl alcohol, the reducing agent contains pyridine borane, and the tosylate salt of azetidine 9 is used. In yet another embodiment, the solvent contains isopropanol, the reducing agent contains pyridine borane, and the tosylate salt of azetidine 9 is used.

[0080] Removal of the PG group can occur at any point during the synthesis of compounds of Formula A, B, and C. In one embodiment, it is removed before the reaction between compounds 8 and 9. If the PG group is methyl, it can be removed using BBr3.

[0081] In general, pharmaceutically acceptable salts of compounds of formula A, B, and C can be formed by treating the free base of formula A, B, and C with a pharmaceutically acceptable acid in a suitable solvent. Formula A, B, and C can be isolated, and the salt or salt form formed in a separate step can be isolated without isolating the neutral substance of formula A, B, and C. The formation of pharmaceutically acceptable salts is well known. See, for example, P. Stahl, et al., HANDBOOK OF PHARMACEUTICAL SALTS: PROPERTIES, SELECTION AND USE, (VCHA / Wiley-VCH, 2002); Gould, PL, "Salt selection for basic drugs," International Journal of Pharmaceutics, 33:201-217 (1986); Bastin, RJ, et al., "Salt Selection and Optimization Procedures for Pharmaceutical New Chemical Entities," Organic Process Research and Development, 4:427-435 (2000); and Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, 66:1-19, (1977). Those skilled in the art will understand that the compounds of formulas A, B, and C described herein can be readily converted to and isolated as pharmaceutically acceptable salts. Examples of useful salts include, but are not limited to, HCl salt, benzenesulfonate (besylate), 4-methylbenzenesulfonate (tosylate), or methylsulfonate (mesylate). In one embodiment, the pharmaceutically acceptable salt is a besylate salt. In another embodiment, the pharmaceutically acceptable salt is a tosylate salt. In yet another embodiment, the pharmaceutically acceptable salt is a mesylate. In another embodiment, the pharmaceutically acceptable salt is an HCl salt.

[0082] In one embodiment, the compound of formula A or a pharmaceutically acceptable salt thereof is a compound of formula B [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, —CF3, or —CH3, and the other is H; and R7 is H or PG.

[0083] In a preferred embodiment, the compound of Form B is: [ka]

[0084] Scheme 4 shows the chiral synthesis of enantiomer B.

[0085] Scheme 4 [ka] Scheme 4 shows the chiral synthesis of enantiomer B. The key step in making the chiral compound is setting the chirality of the alcohol in step C, as this chirality is maintained in steps D, D1, E, and F.

[0086] In Step C, prochiral compound 4 undergoes chiral reduction to selectively form chiral alcohol 6B. During the reduction, dihydroquinoline 5B is also formed. Without wishing to be bound by theory, it is believed that residual Pd in ​​compound 4 (from the Suzuki coupling) leads to over-reduction of compound 6B, forming compound 5B. As shown in Scheme 3, dihydroquinoline 5B can be oxidized to quinoline 6B using at least one oxidizing agent. Examples of suitable oxidizing agents include, but are not limited to, MnO, DDQ, and oxygen. In some embodiments, the oxidizing agent is MnO. In another embodiment, the oxidizing agent is oxygen in an inert gas. While various oxygen concentrations can be used, using lower concentrations is desirable because doing so helps minimize fire hazards. In one embodiment, the oxidizing agent is about 5% oxygen in nitrogen. The mixture of 5B and 6B is typically not isolated prior to the oxidation reaction.

[0087] In one embodiment, a chiral reduction protocol that can be used is the CBS chiral ketone reduction. The CBS reduction utilizes a borolidone reagent, such as trimethyl borate, in combination with a chiral amino compound, such as diphenyl-[(2R)-pyrrolidin-2-yl]methanol, and a boron-based reducing agent, such as BH3-Me2S. Other borolidone reagents that can be used include triethyl borate, triisopropyl borate, tri-t-butyl borate, or tributylborane. Other boron-based reducing agents that can be used include borane N,N-diethylaniline, BH3-THF, or other borane sources. Other reduction protocols, such as enzymatic or catalytic hydrogenation, can also be used.

[0088] Examples of compounds 5B and 6B that can be made using the methods disclosed herein include, but are not limited to: [Table 2-1] [Table 2-2]

[0089] The above compounds are also potential impurities in the compound of formula B, where R7 is H, R1 is H, and R2 is CF3.

[0090] Steps D, E, and F of Scheme 4 can be completed as described in Scheme 3. Preferably, the oxidation, deprotection, cyclization, and coupling reactions maintain the stereochemical integrity of the starting materials, i.e., little, if any, stereochemical purity is lost. For example, if compound 6B is 95±5% single enantiomer (as determined by chiral HPLC or chiral gas chromatography (GC)), the resulting product, compound 7B, is 95±5% single enantiomer (as determined by chiral HPLC or chiral GC). Similarly, if compound 7B or 8B is 97% single enantiomer, compounds 8B and A are also 97% single enantiomer.

[0091] Examples of compounds of formula 7B that can be made using the methods disclosed herein include, but are not limited to, the following: [Table 3]

[0092] The above compounds are also potential impurities in the compound of formula B, where R7 is H, R1 is H, and R2 is CF3.

[0093] Additional potential impurities in compounds of formula B (wherein R7 and R1 are H and R2 is CF3) include the following: [Table 4]

[0094] The dihydroquinoline compound shown above is derived from compound 5B which was not oxidized to compound 6B.

[0095] Still other potential impurities in compounds of formula B (wherein R7 and R1 are H and R2 is CF3) include the following: [Table 5-1] [Table 5-2]

[0096] Scheme 5 shows an alternative method for preparing compound 4. In Scheme 5, compound 1 is reacted with a -CHCH(OR) synthon to form compound 15, which then undergoes Suzuki coupling to form compound 4.

[0097] Scheme 5 [ka] In Scheme 5, Step A can be carried out as described in Scheme 3, Step B, and Scheme 5, Step B can be carried out as described in Scheme 3, Step A. Such rearrangements may require less catalyst and boronic acid, resulting in a more economical process. The rearranged reactions may provide similar quality and yields.

[0098] An alternative method for preparing compound 3 is shown in Scheme 6.

[0099] Scheme 6 [ka] In Scheme 6, R7 is a protecting group or H. Step A is a decarboxylative iodination carried out by reacting 7-methoxy-4-hydroxyquinoline, compound 15, with I2 and K3PO4 in ACN with heating to about 50°C. The resulting product, 3-iodo-7-methoxyquinolin-4-ol (compound 11), is then cross-coupled with compound 2 in Step B. This cross-coupling step can be carried out as described in Scheme 3, Step A. While a variety of conditions can be used (see Scheme 3, Step A), a suitable set of conditions involves the use of either PEPPSI™-IPr or (Amphos)2PdCl2 as the catalyst, K2CO3 as the base, and tert-amyl alcohol / ACN / water or dioxane / water as the solvent.

[0100] The bromination in Step C can be carried out as described in Scheme 1, Step B. Compound 17, shown above, is shown as the bromide. Although not shown in Scheme 6, art-known chlorination or iodination methods could be used to replace the bromide with a chloride or iodide. Phosphorus tribromide or phosphorus oxybromide can be used in the bromination step, although in one embodiment, phosphorus oxybromide is preferred. In an alternative embodiment, the bromination in Step C can be completed prior to the cross-coupling in Step B. In Step D, the Grignard addition can be completed as described in Scheme 1, Step C.

[0101] Alternatively, compound 17 can be treated with a strong base to generate an anion, which can be reacted with aldehyde 13a to give compound 18. Examples of strong bases include tert-BuLi and n-BuLi. When n-BuLi is used as the strong base, 4-butyl-3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-methoxy-quinoline and 3-methoxy-9-(trifluoromethyl)benzo[3,4]cyclobuta[1,2-c]quinoline may be formed as impurities (see structures in the table below).

[0102] Oxidation of the benzyl alcohol of compound 18 (Step E) can be completed as described in Step D of Scheme 1. Deprotection of the hydroxy of compound 3 can occur at any step of the synthesis using a suitable reagent, for example, when PG is methyl, a suitable reagent is BBr3. [Table 6]

[0103] Scheme 7 [ka] Scheme 7 shows an alternative method for preparing compound 3. In Scheme 7, compound 17 is reacted with compound 13, where R3 is Cl or N(OMe)R4 (where R4 = C1-C4 alkyl), and R7 is a protecting group or H. This reaction affords ketone 3 without the intervention of an alcohol.

[0104] Scheme 8 [ka] Scheme 8 shows an alternative method for preparing Compound A. In Scheme 8, Step G, R7 is a protecting group or is H. In Step G, Compound A can be obtained by reaction with an amine in a similar manner as outlined in Scheme 3, Step F.

[0105] In some embodiments, the compound of Formula A, Formula B, or Formula C has a purity of at least 98.1% area, 98.2% area, 98.3% area, 98.4% area, 98.5% area, 98.6% area, 98.7% area, 98.8% area, 98.9% area, 99.0% area, 99.1% area, 99.2% area, 99.3% area, 99.4% area, 99.5% area, or 99.6% area.

[0106] In another embodiment, the compound of formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and H available for reaction with a reducing agent. Suitable reducing agents are disclosed elsewhere in this application.

[0107] In yet another embodiment, the compound of formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, wherein the compound of formula A has at least a 98% area as measured using HPLC. The compound of formula A is prepared using the methods disclosed herein.

[0108] In one embodiment, a compound of formula B, [ka] wherein R1 is H, R2 is -CF3, and R7 is an aldehyde of formula 8b or a hydrated form of aldehyde 8b: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and H accessible by reacting with a reducing agent. Suitable reducing agents are disclosed elsewhere in this application. In one embodiment, the compound of formula B has at least 98% area as measured using HPLC. In another embodiment, the compound of formula B has at least 92% enantiomeric excess as measured using HPLC. In yet another embodiment, the compound of formula B contains a dimer and / or dimer 2. In yet another embodiment, the compound of formula B contains (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. In another embodiment, the compound of formula B contains (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. In one embodiment, the compound of formula B contains (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. In yet another embodiment, the compound of formula B contains less than 1% dihydroquinoline and quinoline-based impurities as determined using HPLC. In one embodiment, the compound of formula B contains less than 1% dihydroquinoline-based impurities as determined using HPLC. In yet another embodiment, the compound of formula B contains less than 1% quinoline-based impurities as determined using HPLC.

[0109] In another embodiment, the compound of formula B, wherein at least one of the following is true: [ka] or a pharmaceutically acceptable salt thereof is disclosed: The compound of formula B is at least 98% area as measured using HPLC; The compound of formula B is in at least 92% enantiomeric excess as determined using HPLC; The compound of formula B contains dimer and / or dimer 2; The compound of formula B contains (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol; The compound of formula B contains (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol; The compound of formula B contains (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol; The compound of formula B contains less than 1% of dihydroquinoline and quinoline-based impurities as determined using HPLC; The compound of formula B contains less than 1% of a dihydroquinoline-based impurity as determined using HPLC; The compound of formula B contains less than 1% of a quinoline-based impurity as determined using HPLC.

[0110] In another embodiment, the compound of formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H, and R7 is H or PG, providing a compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compounds of Formula A, B, and C have an enantiomeric excess of at least about 92%. Suitable reducing agents are disclosed elsewhere in this application.

[0111] In one embodiment, a compound of formula A [ka] or a pharmaceutically acceptable salt thereof, wherein either R1 or R2 is independently Cl, F, -CF3, or -CH3, and the other is H; and R7 is H or PG containing a C3-C7 alcohol. The compound of Formula A is prepared using the methods disclosed herein. The presence and amount of the C3-C7 alcohol is determined using gas chromatography.

[0112] In some embodiments, the C3-C7 alcohol is a C3-C4 alcohol, or a C3-C5 alcohol, or a C3-C6 alcohol, or a C4-C5 alcohol, or a C4-C6 alcohol, or a C4-C7 alcohol, or a C5-C6 alcohol, or a C5-C7 alcohol, or a C6-C7 alcohol. In additional embodiments, the C3-C7 alcohol is a C3 alcohol, or a C4 alcohol, or a C5 alcohol, or a C6 alcohol, or a C7 alcohol. The C3 alcohol is isopropanol, the C4 alcohol is tert-butanol, and the C5 alcohol is tert-amyl alcohol. In one embodiment, the C3-C7 alcohol comprises isopropanol, tert-butanol, and / or tert-amyl alcohol. In one embodiment, the compound of Formula A contains isopropanol. In one embodiment, the compound of Formula A contains tert-butanol. In one embodiment, the compound of Formula A contains tert-amyl alcohol.

[0113] When the compounds of formulae A, B, and C contain a C3-C7 alcohol, the C3-C7 alcohol is present in an amount of 1% by weight or less, or less than or equal to 0.9% by weight, or less than or equal to 0.8% by weight or less, or less than or equal to 0.7% by weight, or less than or equal to 0.6% by weight, or less than or equal to 0.5% by weight, or less than or equal to 0.4% by weight, or less than or equal to 0.3% by weight, or less than or equal to 0.2% by weight, or less than or equal to 0.1% by weight.

[0114] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound comprises (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. The compound of Formula A can be prepared using the methods disclosed herein. When present, (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of 0.5%, or 0.4%, or 0.3%, or less, as measured by HPLC.

[0115] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A contains (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. When present, (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of about 0.5%, or about 0.4%, or about 0.3%, or less, as measured by HPLC.

[0116] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound contains (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. The compound of Formula A can be prepared using the methods disclosed herein. When present, (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of 0.05% to 0.5%, as determined using HPLC.

[0117] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A contains (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. When present, (5R)-5-[4-[2-[3-(chloromethyl)azetidin-1-yl]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of about 0.05% to about 0.5%, as determined using HPLC.

[0118] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound contains (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. The compound of Formula A can be prepared using the methods disclosed herein. When present, (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of 0.01% to 0.5%, as determined using HPLC.

[0119] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A contains (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. When present, (5R)-5-[4-[2-[[2-(chloromethyl)-3-fluoro-propyl]amino]ethoxy]phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol is present at a level of about 0.01% to about 0.5%, as determined using HPLC.

[0120] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and contains the following dimer: [ka]

[0121] The compound of formula A can be prepared using the methods disclosed herein. If present, the dimer is present at a level of about 0.01% to about 0.5%, as determined using HPLC. The dimer can be formed during jet milling of the compound of formula A; i.e., the dimer is formed after the compound of formula A is isolated.

[0122] Also, the compound of formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound contains the following dimer 2: [ka]

[0123] The compound of formula A can be prepared using the methods disclosed herein. When present, dimer 2 is present at a level of about 0.01% to about 0.5%, as determined using HPLC. Dimer 2 can be formed during jet milling of the compound of formula A; i.e., the dimer is formed after the compound of formula A is isolated.

[0124] In some embodiments, the compound of formula A may contain dimer, dimer 2, or a mixture thereof. Preferably, the amount of dimer and / or dimer 2 in the compound of formula A is minimized. More preferably, the amount of dimer and / or dimer 2 in the compound of formula A is undetectable, or even more preferably zero, as measured using HPLC.

[0125] Without wishing to be bound by theory, it is believed that dimer and dimer 2 form from or through interaction with amorphous (i.e., non-crystalline) Formula A. Jet milling is believed to increase the content of amorphous Formula A, thereby increasing dimer and / or dimer 2. After storage under stress conditions of 40°C and 75% relative humidity for 6 months, about 0.2% dimer by HPLC and about 0.2% dimer 2 by HPLC were identified in the compound of Formula A. If dimer and / or dimer 2 are formed before storage, they are present in an amount less than about 0.05% as measured by HPLC. Reducing the content of amorphous Formula A should reduce the amount of dimer and dimer 2 formed.

[0126] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A contains 4-butyl-3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-methoxy-quinoline. If present, 4-butyl-3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-methoxy-quinoline is present at a level of less than about 0.5%, as determined using HPLC.

[0127] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound of structure 8: [ka] or a salt thereof, in a solvent, with an amine of structure 9, [ka] or a salt thereof, and a reducing agent, wherein the compound of Formula A contains 3-methoxy-9-(trifluoromethyl)benzo[3,4]cyclobuta[1,2-c]quinoline. If present, 3-methoxy-9-(trifluoromethyl)benzo[3,4]cyclobuta[1,2-c]quinoline is present at a level of less than about 0.5%, as determined using HPLC.

[0128] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound or pharmaceutically acceptable salt thereof contains 3-methoxy-9-(trifluoromethyl)benzo[3,4]cyclobuta[1,2-c]quinoline in an amount less than about 0.5%, as determined using HPLC.

[0129] Compound of Formula A [ka] or a pharmaceutically acceptable salt thereof, wherein R1 is H, R2 is -CF3, and R7 is H, and the compound or pharmaceutically acceptable salt thereof contains 4-butyl-3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-methoxy-quinoline in an amount of less than about 0.5%, as determined using HPLC.

[0130] All of the methods disclosed herein can be used to prepare enantiomerically enriched compounds of Formula A. Preferably, when an enantiomerically enriched compound of Formula A is prepared to obtain compounds of Formula B and Formula C, the compounds of Formula B and Formula C have an enantiomeric excess of at least about 90%. The enantiomeric excess can be determined using chiral chromatography (such as chiral HPLC) or other methods known in the art. More preferably, the compounds have an enantiomeric excess of at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 97.5%, or at least about 98%, or at least about 98.5%, or at least about 99%, or at least about 99.1%.

[0131] In one embodiment, in any of the previously disclosed compounds of formula A, R7 is H.

[0132] In another embodiment, all of the previously disclosed compounds of formula A have the R-enantiomeric form.

[0133] In yet another embodiment, in any of the previously disclosed compounds of formula A, R 1 is H.

[0134] In still yet another embodiment, in any of the previously disclosed compounds of formula A, R 2 is CF3.

[0135] In one embodiment, in any of the previously disclosed compounds of formula A, R7 is PG. Optionally, PG is methyl.

[0136] In one embodiment, the previously disclosed compound of formula A [ka] or any of its pharmaceutically acceptable salts, wherein either R1 or R2 is independently Cl, F, —CF3, or —CH3, and the other is H, and R7 is H or PG and contains less than 0.15% area of ​​one or more dihydroquinoline or quinoline-based impurities. Dihydroquinoline or quinoline impurities are compounds containing dihydroquinoline and / or quinoline ring systems used to make the compound of Formula A. Intermediates used to prepare or made during the synthesis of the compound of Formula A using the methods disclosed herein are examples of dihydroquinoline and quinoline-based impurities, as are impurities derived from intermediates. In some embodiments, the total amount of dihydroquinoline or quinoline impurities present is less than 1.0% area, or less than 0.9% area, or less than 0.8% area, or less than 0.7% area, or 1.0% area, or less than 0.9% area, or less than 0.8% area, or less than 0.7% area, or less than 0.6% area, or less than 0.5% area, or less than 0.4% area, or less than 0.3% area, or less than 0.2% area, or less than 0.15% area, or less than 0.1% area.

[0137] In one embodiment, the chiral reduction of the ketone group in compound 4 is carried out using a CBS protocol, which includes the use of diphenyl-[(2R)-pyrrolidin-2-yl]methanol, trimethyl borate, and borane N,N-diethylaniline. In a further embodiment, the chiral reduction is carried out in a solvent comprising at least one of THF and 2-MeTHF. Preferably, the resulting chiral alcohol is at least 95% ee as measured using chiral chromatography.

[0138] Also disclosed herein is a process for preparing pyridine borane, which comprises reacting a pyridinium salt with sodium borohydride in a solvent. Various pyridinium salts can be used, with pyridinium p-tosylate being preferred in one embodiment. Solvents that can be used in the preparation of pyridine borane include aprotic solvents. Examples of aprotic solvents include, but are not limited to, ethers. Preferred ethers include tetrahydrofuran and 2-methyltetrahydrofuran. The formation of pyridine borane is typically carried out at a temperature of about 15-30°C or 19-25°C. In one embodiment, the reaction is carried out at room temperature, which is about 19-21°C. The reaction time can vary but is typically about 1-24 hours. In some embodiments, the reaction time is about 2-12 hours. Upon completion, the reaction can be filtered through a filtering agent such as diatomaceous earth. The product can be used without purification or can be purified using methods known in the art.

[0139] The following preparations and examples further illustrate the invention. 1 H NMR can be used to determine purity. 1,3,5-trimethoxybenzene is 1 It can be used as a H NMR reference internal standard.

[0140] Preparation 1 [ka] Scheme 1, Step A: Add DMF (1240 L, 8.0 L / kg) and 7-methoxyquinolin-4-ol (155.04 kg, 885.03 mol) together. Stir at 27.5°C for 30 minutes. Adjust the temperature to 30°C. Add NCS (118.18 kg, 885.03 mol) in portions, maintaining the reaction temperature below 45°C. Adjust the temperature to 40°C. Stir for 24 hours. Cool to 15°C. Add water (4651 L, 30 L / kg). Mix at 25°C for 1 hour, then filter and wash the product with water (775 L, 5 L / kg). Dry under reduced pressure at 65°C to obtain the title compound as a light brown solid (153.34 kg, 82.6% yield).

[0141] Preparation 2 [ka] Scheme 1, Step B: Combine 3-chloro-7-methoxyquinolin-4-ol (154.32 kg, 736.2 mol) and toluene (1543 L, 10 L / kg). Stir at 27.5°C for 30 minutes. Add P(O)Br3 (253.3 kg, 883.39 mol) while maintaining the temperature below 40°C. Heat to 100°C for 3 hours. Cool to 5°C, then add a solution of NaOH (104.94 kg, 2623.6 mol) in water (1312 L, 8.5 L / kg). Concentrate under reduced pressure to 8.5 L / kg while maintaining the temperature below 70°C. Add water (308.6 L, 2.0 L / kg). Concentrate under reduced pressure below 70°C to 8.5 L / kg. Add water (308.6 L, 2.0 L / kg). Concentrate under reduced pressure below 70°C to 8.5 L / kg. Cool to 27.5°C. Add water (1543 L, 10 L / kg). Cool to 5°C. Filter the product and wash with water (308.6 L, 2.0 L / kg). Add a solution of NaOH (15.43 kg, 385.8 mol) in water (771.6 L, 5.0 L / kg) and stir at 30°C for 5 minutes. Filter the solid and wash with water (463.0 L, 3.0 L / kg). Dry under reduced pressure at 65°C to obtain the title compound as a brown solid (174.20 kg, 86.8% yield).

[0142] Alternative preparation 2 4-Bromo-3-chloro-7-methoxyquinoline Scheme 1, Step B: Add 3-chloro-7-methoxyquinolin-4-ol (25.0 g, 119 mmol) and DMF (225 mL, 9 mL / g) together. Cool to 0 °C. Add PBr3 while maintaining the temperature below 10 °C. Heat to 47.5 °C. Stir for 3 h. Cool to 25 °C. Add a solution of 20 wt% Na2CO3 (250 mL). Filter the solid and wash with water (125 mL). Dry under reduced pressure to give the title compound as a brown solid (29.0 g, 89% yield).

[0143] Preparation 3 [ka] Scheme 6, Step A: 4-Hydroxy-7-methoxyquinoline-3-carboxylic acid (10.00 g, 43.34 mmol), KPO (11.3 g, 52.2 mmol), and ACN (100 mL) are added together. Iodine (22.0 g, 86.7 mmol) is added and heated at 50° C. for 16 hours. The product is filtered, washed with ACN, and dried under reduced pressure at 40° C. to give the title compound (9.70 g, 72.1% yield). 1 H NMR (400 MHz, d6-DMSO) δ 11.98 (s, 1H), 8.48 - 8.35 (m, 1H), 8.07 - 7.95 (m, 1H), 7.01 - 6.94 (m, 2H), 3.86 (s, 3H). 13 C NMR (101 MHz, d6-DMSO) δ 172.97, 162.36, 144.76, 141.78, 127.89, 117.41, 114.75, 99.55, 81.11, 55.98.

[0144] Preparation 4 [ka] Scheme 6, Step B: 3-Iodo-7-methoxy-quinolin-4-ol (5.00 g, 16.6 mmol), 2-fluoro-4-(trifluoromethyl)phenylboronic acid (7.27 g, 33.2 mmol), [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride (0.289 g, 0.416 mmol), potassium carbonate (4.87 g, 34.9 mmol) are added to the reaction mixture. The mixture is degassed with nitrogen, followed by the addition of tert-amyl alcohol (50 mL) and water (10 mL). Heat to 70°C for 3 hours, then add additional 2-fluoro-4-(trifluoromethyl)phenylboronic acid (1.82 g, 8.32 mmol), [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride (0.058 g, 0.083 mmol) and stir for 1 hour. Cool to 22°C, then filter and wash with 1:5 water:tert-amyl alcohol, then water. Dry under reduced pressure at 45°C to give the title compound (4.26 g, 66.2% yield).

[0145] Preparation 5 [ka] Scheme 1, Step D: Add 2.0 M isopropylmagnesium chloride in THF (239 m, 477 mmol) to a stirred solution of 4-bromo-3-chloro-7-methoxyquinoline (100 g, 367 mmol) in THF (1.5 L) at −25° C. and maintain the temperature below −20° C. Stir at −25° C. for 1 h. Add 4-fluorobenzaldehyde (54.65 g, 440 mmol) slowly, maintaining the temperature below −5° C. Stir at −10° C. for 2 h. Add water (500 mL) and EtOAc (500 mL). Stir at 27.5° C. for 30 min. Add 1 M aqueous HCl (500 mL). Stir at 27.5° C. for 30 min. Separate the organic layer and wash with 5% aqueous NaCl (500 mL) and 5% aqueous NaHCO (500 mL). Distill under reduced pressure below 50°C until 4.5 mL / g remains. Add EtOAc (500 mL) and distill under reduced pressure below 50°C until 5.5 mL / g remains. Add EtOAc (1200 mL). After 30 min at 27.5°C, add water (500 mL) and KBr (8.73 g, 0.073 mol), followed by TEMPO (2.81 g, 0.18 mol). After 30 min at 27.5°C, cool to -2.5°C and add an aqueous solution of sodium hypochlorite prepared by mixing sodium hypochlorite (500 mL, 74.44 mmol) with a solution of NaHCO3 (25.0 g in 500 mL of water). Stir the mixture at -2.5°C for 1 h, warm the solution to 27.5°C, separate the organic layer, and extract the aqueous layer with EtOAc (500 mL). Wash the combined organics with water (2 x 500 mL), treat with neutral activated carbon (grade PF-33N, 5 g, 0.05 w / w), and filter through a bed of Hyflo. Distill under reduced pressure below 50 °C until 4.5 mL / g remains. Solvent exchange to EtOH by adding EtOH (500 mL) and distill until 4.5 mL / g remains (repeat co-distillation / concentration 1x). Heat to 75 °C and stir for 30 min. Cool slowly to 12.5 °C. Stir for 30 min, filter the solid, and wash with EtOH (100 mL). Dry under reduced pressure at 42.5 °C to give the title compound as a brown solid (78 g, 67.8% yield).

[0146] Alternative preparation 5 (3-chloro-7-methoxyquinolin-4-yl)-(4-fluorophenyl)methanone Scheme 2, Step C: Add 2.0 M isopropylmagnesium chloride in THF (119 mL, 0.238 mol) dropwise to a stirred solution of 4-bromo-3-chloro-7-methoxyquinoline (50 g, 0.183 mol) in THF (750 mL) at −25° C., maintaining the temperature below −15° C. Stir at −22.5° C. for 2 h. Add bis[2-(N,N-dimethylamino)ethyl]ether (35.28 g, 0.220 mol) and stir for 30 min. Slowly add 4-fluorobenzoyl chloride (34.8 g, 0.220 mol), maintaining the temperature below −5° C. Stir at −7.5° C. for 1 h. Filter through diatomaceous earth and wash with EtOAc (250 mL). The combined filtrates are washed with 1 M aqueous HCl (250 mL), followed by 5% aqueous NaCl (250 mL). Treat with neutral activated carbon (grade PF-33N, 5 g, 0.05 w / w) and filter through diatomaceous earth. Distill under reduced pressure below 50°C until 4.5 mL / g remains. Exchange the solvent for EtOH by adding EtOH (500 mL) and distilling (co-distillation / concentration 1x) until 4.5 mL / g remains. Heat to 75°C and stir for 30 minutes. Cool slowly to 12.5°C, stir for 30 minutes, filter, and wash the solid with EtOH (100 mL). Dry under reduced pressure at 42.5°C to obtain the title compound as a brown solid (38 g, 66% yield).

[0147] Preparation 6 [ka] 2 M isopropylmagnesium chloride in THF (2.8 mL, 5.6 mmol) is added to a solution of 4-bromo-3-chloro-7-methoxy-quinoline (0.74 g, 2.7 mmol) in THF (12 mL) at −40° C. and stirred for 1 h. A solution of 2,3,5,6-tetradeuterio-4-fluoro-benzoyl chloride (0.97 g, 5.97 mmol) in THF (1.5 mL) is added, and the mixture is stirred at −40° C. for 30 min, then warmed to room temperature for 30 min. A 2:1 mixture of water and saturated NH4Cl (3 mL) is added to the reaction. The mixture is diluted with water (40 mL) and extracted with DCM (3 × 20 mL). The organic layers are combined, washed with saturated brine (10 mL), dried over sodium sulfate, and filtered. The filtrate is concentrated onto silica gel (8 g) and purified by silica chromatography eluting with a gradient of 10-30% EtOAc in hexanes to afford the title compound (0.48 g, 55% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1H), 7.57 (d, J = 2.6 Hz, 1H), 7.40 (d, J = 9.2 Hz, 1H), 7.31 (dd, J = 9.2, 2.6 Hz, 1H), 3.95 (s, 3H).

[0148] Preparation 7 [ka] Cleavage of the methyl ether: Add 1 M BBr3 in DCM (112.0 g, 0.45 mol) to a stirred solution of (3-chloro-7-methoxyquinolin-4-yl)(4-fluorophenyl)methanone (47.0 g, 0.15 mol) in DCM (494 mL) at 15 °C, maintaining the temperature below 30 °C. Warm to 35 °C and stir for 50-68 h. Quench the reaction with water (47 mL) and adjust the pH to 11-12 using 2 M NaOH (aq). Separate the layers and adjust the pH of the aqueous layer to 7-8 using 2 M HCl (aq) to precipitate the product. Filter the solid, wash with water and DCM, and dry under reduced pressure to give the title compound (39.4 g, 85.4% yield).

[0149] Alternative preparation 7 (3-chloro-7-hydroxyquinolin-4-yl)(4-fluorophenyl)methanone Cleavage of the methyl ether: Add (3-chloro-7-methoxyquinolin-4-yl)(4-fluorophenyl)methanone (80 g, 0.25 mol), HBr 48% (800 mL), and acetic acid (400 mL). Stir at 30° C. for 30 minutes. Heat to 110° C. for 48 hours. Cool to 90° C. and distill under reduced pressure below 100° C. until 7.5 mL / g remains. Cool to 5° C. and add water (800 mL), maintaining the temperature below 25° C. Stir at 5° C. for 2 hours, then filter and wash with water (80 mL). Treat the solid with NaHCO3 (aq) (24 g, 0.30 w / w in 1200 mL, 15 mL / g water) at 25° C. for 30 minutes. Filter and wash the solid with water (160 mL). Drying under reduced pressure at 60°C gives the title compound as a brown solid (72g, 94.2% yield, 98.92% purity, determined by the following assay conditions: Shimadzu LC-20A HPLC system, Agilent Bonus RP column (75mm*4.6mm, 3.5mm), column temperature 30°C, gradient of 75% A (0.05% TFA in water) / 25% B (0.05% TFA in AcCN) to 40% A / 60% B over 20 minutes, then 5% A / 95% B over 2 minutes, flow rate 1.5mL / min, elution with UV 245nm).

[0150] Preparation 8 [ka] Add 1 M boron tribromide in DCM (10 mL, 10 mmol, 3.3 equiv.) to a solution of (3-chloro-7-methoxy-4-quinolyl)-(2,3,5,6-tetradeuterio-4-fluoro-phenyl)methanone (0.96 g, 3.0 mmol) in DCM (24 mL) and heat at 35 °C for 24 h. Add a second aliquot of 1 M boron tribromide in DCM (10 mL, 10 mmol) and heat at 35 °C for 2 days. Cool to room temperature, pour into saturated sodium bicarbonate (60 mL), and extract with DCM (2 × 60 mL). Dry the combined organic layers over sodium sulfate, filter, and concentrate onto silica gel. Purify by silica chromatography eluting with a gradient of 0–8% MeOH in DCM to give the title compound (0.84 g, 92% yield) as a tan solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.55 (s, 1H), 8.89 (s, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.35 (d, J = 9.0 Hz, 1H), 7.21 (dd, J = 9.1, 2.5 Hz, 1H).

[0151] Preparation 9 [ka] Scheme 3, Step A: Under a nitrogen atmosphere, (3-chloro-7-hydroxy-4-quinolyl)-(4-fluorophenyl)methanone (500.0 g, 1.66 mol), 2-fluoro-4-(trifluoromethyl)phenylboronic acid (447.9 g, 2.15 mol), NaHCO (210.0 g, 2.49 mol), THF (3 L), and water (750 mL) are combined. The mixture is degassed with nitrogen, and Pd-XPhos-G (13.0 g, 16.6 mmol) is added. The reaction mixture is then heated at 65 °C for 18.0 h. The mixture is cooled to 20–30 °C and filtered through diatomaceous earth (approximately 50 g), and the filter cake is rinsed with EtOAc (1.5 L). The combined organic layers are separated and concentrated with the addition of EtOAc (3×1.5 L) to give an EtOAc solution (2.5 L) and washed with 7% aqueous NaHCO (2 L) and 25% aqueous NaCl (2 L). The organic layer is treated with silica thiol (50 g) at 55° C. for 2 hours. The mixture is filtered through diatomaceous earth (approximately 50 g), washed with EtOAc (1.5 L), concentrated to 1.5 L, cooled to 25° C., and n-heptane (7.5 L) is added over 2 hours. The resulting mixture is cooled to 5° C., stirred for 1 hour, filtered, washed with n-heptane (1.5 L), and dried at 45° C. for 18 hours to give the title compound as an off-white solid (660.0 g, 88.0% yield, 94.9% purity, as determined by the assay described in Alternative Preparation 7). 1 H NMR (500 MHz, d6-DMSO) δ 10.60 (s, 1H), 8.89 (d, J = 1.3 Hz, 1H), 7.74 - 7.66 (m, 3H), 7.59 - 7.44 (m, 4H), 7.29 - 7.20 (m, 3H). 13C NMR (126 MHz, d6-DMSO) δ 193.97, 165.61 (d, J = 255.4 Hz), 159.66, 159.01 (d, J = 247.8 Hz), 150.91, 149.34, 143.11, 133.40 (d, J = 2.8 Hz), 132.78 (d, J = 2.6 Hz), 132.33 (d, J = 9.9 Hz), 130.91 (qd, J = 33.1, 8.1 Hz), 128.39 (d, J = 16.0 Hz), 126.62, 123.08 (qd, J = 272.1, 2.4 Hz), 121.97, 121.35 (m), 121.10 (d, J = 2.9 Hz), 117.63, 116.27 (d, J = 22.3 Hz), 113.22 (dq, J = 25.8, 3.8 Hz), 110.70.

[0152] Preparation 10 [ka] Degass the mixture under vacuum and recharge with nitrogen (3x) with a solution of (3-chloro-7-hydroxy-4-quinolyl)-(2,3,5,6-tetradeuterio-4-fluorophenyl)methanone (0.83 g, 2.6 mmol), [2-fluoro-4-(trifluoromethyl)phenyl]boronic acid (0.82 g, 3.9 mmol), XPhos Pd G2 (82 mg, 4 mol%), potassium carbonate (1.08 g, 7.8 mmol), water (5 mL), and tert-amyl alcohol (15 mL). Heat to 80 °C for 1 h, cool to room temperature, dilute with water (40 mL), and extract with EtOAc (2 x 40 mL). Combine the organic layers, wash with saturated brine (20 mL), dry over sodium sulfate, filter, and concentrate under reduced pressure. The residue was dissolved in DCM and concentrated onto silica gel. Purify on silica gel chromatography, eluting with a gradient of 20-25% EtOAc in hexanes to give the title compound (0.96 g, 85% yield) as a solid. 1H NMR (400 MHz, DMSO-d6) δ 10.59 (s, 1H), 7.71 (dd, J = 10.0, 1.7 Hz, 1H), 7.61 - 7.49 (m, 2H), 7.49 - 7.41 (m, 2H), 7.23 (dd, J = 9.1, 2.5Hz, 1H), 19 F NMR (376 MHz, DMSO-d6) δ -61.30 (s), -103.61 (s), -111.80 (m).

[0153] Preparation 11 [ka] Scheme 5, Step A: Under a nitrogen atmosphere, 20% potassium tert-butoxide in THF solution (92.98 g, 165.7 mmol) is added over 40 minutes to a solution of 2,2-diethoxyethanol (11.56 g, 86.2 mmol) and THF (60 mL) at 5° C. and stirred for 0.5 hours. In a separate reactor, (3-chloro-7-hydroxy-4-quinolyl)-(4-fluorophenyl)methanone (20 g, 66.3 mmol) and THF (160 mL) are added, and the resulting solution is added to the potassium tert-butoxide / 2,2-diethoxyethanol / THF mixture at 5° C. After 16 hours, 20% aqueous citric acid (60 mL) and 20% aqueous NaCl (60 mL) are added to the reaction, the organic phase is separated, and activated carbon (1.5 g) is circulated at 25° C. for 0.5 hours. The organic phase is concentrated with the addition of MTBE (3 x 100 mL) to give a solution in MTBE (160 mL) and washed with 20% aqueous NaCl (60 mL). The resulting organic layer is concentrated to 60 mL, heated to 40 °C, and n-heptane (80 mL) is added over 1 h. The resulting slurry is stirred at 40 °C for 1 h, cooled to 5 °C over 2 h, and stirred for an additional 16.0 h. The solid is filtered, washed with n-heptane (40 mL), and dried at 45 °C to give the title compound as an off-white solid (21.0 g, 76.2% yield). 1H NMR (500 MHz, d6-DMSO) δ 10.55 (s, 1H), 8.85 (s, 1H), 7.73 (d, J = 8.5 Hz, 2H), 7.41 (d, J = 2.5 Hz, 1H), 7.34 (d, J = 9.0 Hz, 1H), 7.21 (dd, J = 9.1, 2.5 Hz, 1H), 7.10 (d, J = 8.6 Hz, 2H), 4.80 (t, J = 5.1 Hz, 1H), 4.06 (d, J = 5.2 Hz, 2H), 3.64 (dq, J = 9.5, 7.0 Hz, 2H), 3.53 (dq, J = 9.5, 7.0 Hz, 2H), 1.10 (t, J = 7.1 Hz, 6H)

[0154] Alternative preparation 11 (3-chloro-7-hydroxyquinolin-4-yl)(4-(2,2-diethoxyethoxy)phenyl)methanone Scheme 5, Step A: A reactor was charged with (3-chloro-7-hydroxyquinolin-4-yl)(4-(2,2-diethoxyethoxy)phenyl)methanone (992.7 g, 2.39 mol), (2-fluoro-4-(trifluoromethyl)phenyl)boronic acid (650 g, 3.10 mol), NaHCO3 (304 g, 3.58 mol), THF (6.0 L), and water (1.5 L). The solution was degassed with nitrogen. PdXPHos G2 (14.40 g, 17.9 mmol) was added. Stir at 65 °C for 16 h. Cool to 30 °C and add water (1.0 L). Filter through diatomaceous earth and wash with THF (3.0 L). Add water (2.0 L) and heptane (2.0 L). The organic layer was separated, treated with CUNO (activated carbon filter media), and crystallized from heptane to give the title product (1177 g, 89.54% yield, 98.54% purity, determined using the following HPLC conditions: Shimadzu LC-20A HPLC system, Agilent Bonus RP column (75 mm*4.6 mm, 3.5 mm), column temperature 30°C, gradient of 75% A (0.05% TFA in water) / 25% B (0.05% TFA in AcCN) to 40% A / 60% B over 20 minutes, then 5% A / 95% B over 2 minutes, flow rate of 1.5 mL / min, elution with UV at 245 nm).

[0155] Preparation 12 [ka] Scheme 3, Step B: Under a nitrogen atmosphere, 1 M potassium tert-butoxide in THF (291 mL, 0.29 mol) is added to a solution of 2,2-diethoxyethanol (20.3 g, 0.15 mol) in THF (150 mL) over 1.0 h at 5° C. In a separate reactor, (3-(2-fluoro-4-(trifluoromethyl)phenyl)-7-hydroxyquinolin-4-yl)(4-fluorophenyl)methanone (50.0 g, 0.12 mol) is dissolved in THF (200 mL), and the resulting solution is added to the potassium tert-butoxide / 2,2-diethoxyethanol / THF mixture over 40 min at 5° C. After 16.0 h, 20% citric acid (150 mL) is added to the reaction mixture, and the organic phase is washed with 25% aqueous NaCl (150 mL). The organic layer is circulated through activated carbon (1.5 g) at 25° C. for 2.0 hours, then concentrated to remove THF while MTBE (2×250 mL) is added to produce a final volume of 400 mL of MTBE solution. The MTBE solution is washed with 20% brine (150 mL), concentrated to a final volume of 150 mL, heated to 40° C., and n-heptane (400 mL) is added over 2.0 hours. The resulting slurry is stirred at 40° C. for 1.0 hour, cooled to 5° C. over 3.0 hours, and stirred for 16.0 hours. The solid is filtered, washed with n-heptane (100 mL), and dried at 55° C. for 16.0 hours to give the title compound as a yellow solid (51.1 g, 80.7% yield, 94.3% purity, determined using the following HPLC conditions: Shimadzu LC-20A HPLC system, Agilent Bonus RP column (75 mm*4.6 mm, 3.5 mm), column temperature 30° C., 65% A (0.05% TFA in water) / 35% B (0.05% TFA:MeOH in AcCN=70:30 (v / v) to 25% A / 75% B gradient over 20 minutes, then 5% A / 95% B over 5 minutes, flow rate 1.5 mL / min, UV 245 nm elution). 1H NMR (500 MHz, d6-DMSO) δ 10.65 (s, 1H), 8.89 (d, J = 1.5 Hz, 1H), 7.73 - 7.67 (m, 1H), 7.61 - 7.49 (m, 4H), 7.49 - 7.42 (m, 2H), 7.22 (dd, J = 9.1, 2.5 Hz, 1H), 7.01 - 6.96 (m, 2H), 4.77 (t, J = 5.1 Hz, 1H), 4.01 (d, J = 5.2 Hz, 2H), 3.64 (dq, J = 9.6, 7.0 Hz, 2H), 3.52 (dq, J = 9.6, 7.0 Hz, 2H), 1.09 (t, J = 7.0 Hz, 6H). 13 C NMR (126 MHz, d6-DMSO) δ 193.48, 163.20, 159.70, 159.04 (d, J = 248.5 Hz), 150.77, 149.02, 144.13, 133.38 (d, J = 3.0 Hz), 131.71, 131.36 - 130.31 (m), 129.18, 128.50 (d, J = 16.0 Hz), 126.78, 123.06 (qd, J = 272.5, 2.3 Hz), 121.43 - 121.14 (m), 121.03, 120.92, 117.90, 114.97, 113.25 (dq, J = 25.4, 3.6 Hz), 110.44, 99.65, 68.31, 62.02, 15.19.HRMS(ESI)m / z:[M+H]+C 29 H 26 Calculated value of F4NO5: 544.1681, measured value: 544.1669.

[0156] Alternative preparation 12 (4-(2,2-diethoxyethoxy)phenyl)(3-(2-fluoro-4-(trifluoromethyl)phenyl)-7-hydroxyquinolin-4-yl)methanone Scheme 5, Step B: Under a nitrogen atmosphere, (3-chloro-7-hydroxyquinolin-4-yl)(4-(2,2-diethoxyethoxy)phenyl)methanone (10.0 g, 24.1 mmol) is combined with [2-fluoro-4-(trifluoromethyl)phenyl]boronic acid (6.5 g, 31.3 mmol), NaHCO (3.0 g, 36.1 mmol), THF (60 mL), and water (15 mL). The resulting mixture is degassed with nitrogen, PdXPhos-G (0.2 g, 0.24 mmol) is added, and the mixture is heated to 65 °C for 16.0 h. The mixture is cooled to 25 °C, filtered through diatomaceous earth (10 g), and rinsed with EtOAc (30 mL). The combined organic streams are concentrated with the addition of EtOAc (3 × 50 mL) to give a 50 mL EtOAc solution, then washed with 7% aqueous NaHCO3 (30 mL) and 25% aqueous NaCl (40 mL). The organic layer is treated with silica thiol (1 g, 5-10% w / w) at 60 °C for 16.0 h. The mixture is cooled to 25 °C, filtered through diatomaceous earth (10 g), and rinsed with EtOAc (30 mL). The organic streams are combined and concentrated with the addition of MTBE (2 × 50 mL) to give a 30 mL MTBE solution. n-Heptane (80 mL) is added over 0.5 h at 40 °C to form a slurry, cooled to 5 °C, and stirred for 1.0 h. The solid is filtered, washed with n-heptane (20 mL), and dried at 50 °C to give the title compound as an off-white solid (11.0 g, 84% yield).

[0157] Preparation 13 [ka] Scheme 4, Step C: Under a nitrogen atmosphere, (4-(2,2-diethoxyethoxy)phenyl)(3-(2-fluoro-4-(trifluoromethyl)phenyl)-7-hydroxyquinolin-4-yl)methanone (100.0 g, 0.18 mol), diphenyl-[(2R)-pyrrolidin-2-yl]methanol (14.0 g, 55 mmol), trimethyl borate (6.7 g, 64.0 mmol), and THF (1.0 L) are combined. The resulting mixture is stirred at 25° C. for 1.0 h. To the mixture is added 10 M BH-MeS solution (56.0 g, 0.74 mol), and then warmed to 45° C. for 2.0 h. After cooling to 25°C, MeOH (100 mL) is added over 0.5 h, followed by the addition of ethanolamine (23.5 g, 0.37 mol) and heating to 70°C for 16.0 h. The mixture is concentrated to 500 mL, MTBE (500 mL) is added, and the mixture is washed with 25% NaCl (2 x 300 mL). The organic layer is concentrated under vacuum at 40°C while adding THF (3 x 300 mL) to produce a 1.0 L THF solution. MnO2 (32.0 g, 0.36 mol) is added to the THF solution and heated to 60°C for 16.0 h. The solids are removed by filtration through diatomaceous earth (20 g), and the cake is rinsed with THF (500 mL). The combined organic solution is circulated over activated carbon (1.5 g) at 25°C for 2 h. The resulting solution is concentrated under vacuum at 50°C while adding ACN (3 x 300 mL) to give the title compound as an ACN solution (332.5 g of ACN solution containing 96.0 g of the title compound, 96% yield, 97.3% ee). Chiral analysis conditions: Agilent 1260 HPLC system, Chiralpak IG (250 mm * 4.6 mm, 5 μm), column temperature 45°C, 55% water and 45% ACN, 25 min at 40°C, flow rate 1.0 mL / min, UV 230 nm, t (R) Eluting at 16.2 minutes. NMR data collected at 100°C to give more highly merged resonances of the atropisomers: 1H NMR (500 MHz, d6-DMSO, 100 °C) δ 8.63 (s, 1H), 8.24 (d, J = 9.3 Hz, 1H), 7.79 - 7.64 (m, 3H), 7.39 - 7.36 (m, 1H), 7.12 (d, J = 8.4 Hz, 2H), 7.09 - 7.03 (m, 1H), 6.84 (d, J = 8.7 Hz, 2H), 6.03 (s, 1H), 4.77 (t, J = 5.1 Hz, 1H), 3.94 (dd, J = 5.0, 1.6 Hz, 2H), 3.83 - 3.45 (m, 4H), 1.32 - 0.83 (m, 6H). 13 C NMR (126 MHz, d6-DMSO, 100 °C) δ 159.62 (d, J = 247.3 Hz), 158.41, 157.30, 150.93, 150.62, 146.46, 136.15, 133.78, 130.81 (qd, J = 33.1, 7.7 Hz), 130.36 (d, J = 16.8 Hz), 128.96, 126.67, 123.58, 123.32 (qd, J =272.4, 2.7 Hz), 121.43 - 120.60 (m), 119.62, 118.80, 114.48, 113.03 (dd, J = 26.0, 3.9 Hz), 110.84, 100.24, 70.30, 68.93, 61.93, 15.12.MS (ESI) m / z: [M+ H]+ 546.2.

[0158] Alternative preparation 13 (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol Scheme 4, Step C: Add diphenyl-[(2R)-pyrrolidin-2-yl]methanol (47 mg, 0.186 mmol), THF (10 mL), and trimethyl borate (1.26 mL, 11.1 mmol). Heat to 35°C for 1 hour. Cool to 25°C and add borane N,N-diethylaniline (2.70 mL, 14.7 mmol). Add a solution of [4-(2,2-diethoxyethoxy)phenyl]-[3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-hydroxy-4-quinolyl]methanone (2.00 g, 3.68 mmol) in THF (10 mL). Heat to 55°C for 80 minutes. Cool to 0°C, then add MeOH (3.00 mL). Heat to 60°C for decomplexation. The title compound was obtained in 98.22% ee. Chiral analysis conditions: Chiralpak IG (250 mm × 4.6 mm, 5 μm), column temperature 45 °C, gradient of 63% to 60% water in ACN over 55 min at 40 °C, flow rate of 1.0 mL / min, UV 230 nm, t (R) Elutes at 37.76 minutes.

[0159] Preparation 14 [ka] TEA (1.8 mL, 12.9 mmol) is added to a suspension of 2-[3-(fluoromethyl)azetidin-1-yl]ethanol.HCl (1.15 g, 6.7 mmol, 3.3 equiv.) in MeOH (10 mL) and stirred for 15 min before diluting with THF (20 mL). The mixture is concentrated under reduced pressure. The residue is suspended in THF (20 mL), filtered, and the solid is washed with THF (5 mL). The filtrate is concentrated under reduced pressure. The residue is diluted with THF (25 mL) and filtered through a cellulose syringe filter. The filtrate is treated with potassium tert-butoxide (1.55 g, 13.8 mmol) and stirred for 10 min. A solution of [3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-hydroxy-4-quinolyl]-(2,3,5,6-tetradeuterio-4-fluoro-phenyl)methanone (0.88 g, 2.0 mmol) in THF (15 mL) is added and the mixture is stirred at room temperature overnight. The reaction is diluted with DCM (100 mL) and washed with half-saturated ammonium chloride (50 mL). The aqueous washes are extracted with DCM (2 × 25 mL). The combined organic layers are washed with saturated brine (25 mL), dried over sodium sulfate, filtered, and concentrated onto silica gel. Purification on silica gel, eluting with a gradient of 0–6% MeOH in DCM, affords the title compound (0.66 g, 60% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 8.86 (d, J = 1.6 Hz, 1H), 7.72 (dd, J = 10.1, 1.7 Hz, 1H), 7.59 - 7.54 (m, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.45 - 7.39 (m, 2H), 7.20 (dd, J = 9.0, 2.6 Hz, 1H), 4.54 (d, J = 6.2 Hz, 1H), 4.42 (d, J = 6.2 Hz, 1H), 3.96 (t, J = 5.5 Hz, 2H), 3.28 (dd, J = 7.7, 1.5 Hz, 2H), 2.97 (dd, J = 7.3, 5.9 Hz, 2H), 2.78 - 2.62 (m, 3H), 19F NMR (376 MHz, DMSO-d6) δ 17.28 (m), -61.24(s), -111.93 (t, J = 8.6 Hz).

[0160] Preparation 15 [ka] Sodium borohydride (0.20 g, 5.2 mmol) was added to a solution of [3-[2-fluoro-4-(trifluoromethyl)phenyl]-7-hydroxy-4-quinolyl]-[2,3,5,6-tetradeuterio-4-[2-[3-(fluoromethyl)azetidin-1]-yl]ethoxy]phenyl]methanone (0.28 g, 0.51 mmol) in a 1:1 mixture of THF and 2-propanol (24 mL). After 24 h (0.20 g, 5.2 mmol) and 72 h (0.20 g, 5.2 mmol), additional sodium borohydride was added. After 6 days, the reaction was diluted with 1 M HCl (20 mL) and DCM (50 mL) and stirred for 10 min. The mixture was diluted with saturated sodium bicarbonate (50 mL). The layers were separated and the aqueous layer was extracted with DCM (2 x 50 mL). The organic layers were combined and concentrated under reduced pressure. The residue was dissolved in MeOH (20 mL), 12 M HCl (2 mL) was added, and heated at 60 °C for 2 h. The mixture was cooled, the pH was adjusted to basic with saturated sodium bicarbonate (50 mL), and extracted with DCM (3 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated onto silica gel. Purification on silica gel, eluting with a gradient of 3-8% MeOH in DCM, afforded the title compound (0.17 g, 63%) as a yellow solid.

[0161] Preparation 16 [ka] Dissolve (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol (236 mg, 43 mmol) in EtOAc (3 mL) with stirring at 500 rpm to give a yellow solution. Add HCl (500 μL, 1 M in EtOAc) to give a clear solution. Stir at 150 rpm for 30 minutes to give a thick slurry after 15 minutes. Filter and collect the off-white precipitate to give the title compound (230 mg, 91.4% yield).

[0162] X-ray powder diffraction (XRPD) of crystalline forms XRPD patterns of crystalline solids were obtained on a Bruker D8 Endeavor X-ray powder diffractometer equipped with a CuKα (1.5418 Å) source and a Linxeye detector, operating at 40 kV and 40 mA. Samples were scanned from 4 to 42° 2θ using a 0.3° primary slit aperture and a 3.9° PSD aperture, with a step size of 0.009° 2θ and a scan rate of 0.5 s / step. Dry powder was loaded into a quartz sample holder, and a glass slide was used to obtain a smooth surface. Diffraction patterns of crystalline forms were collected at ambient temperature and relative humidity. Crystalline peak positions of MDI-Jade were determined after shifting the entire pattern based on an internal NIST 675 standard with peaks at 8.853 and 26.774° 2θ. It is well known in the field of crystallography that for any given crystalline form, the relative intensities of diffraction peaks can vary due to preferred orientation resulting from factors such as crystal morphology and crystal habit. When preferred orientation effects exist, peak intensities change, but the characteristic peak positions of the polymorph remain unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. Furthermore, it is well known in the field of crystallography that for any given crystalline form, angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In the present invention, a peak position variability of ±0.2 2θ° is estimated to account for these potential variations without precluding unambiguous identification of the indicated crystalline form. Confirmation of a crystalline form can be based on any unique combination of prominent peaks.

[0163] A prepared sample of the (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol HCl form is characterized by an XRPD pattern using CuKα radiation having the diffraction peaks (2-theta values) set forth in Table 1, and in particular a peak at 5.6 in combination with one or more peaks selected from the group consisting of 11.1, 20.2, and 22.2.

[0164] Table 1. XRPD peaks of crystalline (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol HCl forms [Table 7]

[0165] Preparation 17 [ka] Scheme 4, Step D: Under a nitrogen atmosphere, (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol (total weight 332.5 g = 96.0 g of the compound of Preparation 13; 94.6% ee; 176.0 mmol, approximately 400 mL) is combined with CsCO (230.0 g, 706.0 mmol) and ACN (560 mL), and the resulting mixture is degassed with nitrogen. The reaction mixture is stirred at 85 °C for 3 h, cooled to 25 °C, filtered through diatomaceous earth (40 g), and the cake is rinsed with ACN (300 mL). The combined filtrates are rinsed and circulated through activated carbon (1.5 g) at 20 °C for 2.0 h. The mixture is concentrated, and during concentration, 2-Me THF (3 x 500 mL) is added to give a 2-Me THF solution (final volume 800 mL). Wash with 1 M KHSO (2 x 500 mL) and 10% aqueous NaCl (300 mL). The resulting organic layer is concentrated to a final volume of 200 mL, and the resulting mixture is warmed to 40 °C. n-Heptane (100 mL) is added dropwise to the mixture and stirred for 1 hour, then additional n-heptane (1.4 L) is slowly added. The resulting slurry is cooled to 5 °C over 3 hours and stirred for 18 hours. The solid is filtered, washed with n-heptane (300 mL), and then dried under vacuum at 45 °C for 24.0 hours to give the title compound as a yellow solid (102.3 g, 95.2% ee; 86.6% yield, 78.3% w / w purity). The purity was determined using the following HPLC conditions: Shimadzu LC-20A HPLC system, Agilent Bonus RP column (75mm*4.6mm, 3.5mM), column temperature 30°C, elution with a gradient of 65% A (0.05% TFA in HO) / 35% B (0.05% TFA in MeOH) to 5% A / 95% B over 30 minutes, flow rate 1.5mL / min, UV 260nm. Chiral analysis conditions: Shimadzu LC-20A HPLC system, Chiralpak IG (250mm*4.6mm, 5µm), column temperature 45°C, elution with a gradient of 55-75% ACN in water over 23 minutes, flow rate 1.2mL / min, UV 268nm to obtain the title compound. (R) 15.9 minutes, 1H NMR (500 MHz, d6-DMSO) δ 11.38 (s, 1H), 9.71 (s, 1H), 8.40 (d, J = 8.1 Hz, 1H), 7.99 (d, J = 9.3 Hz, 1H), 7.65 (d, J = 2.3 Hz, 1H), 7.47 - 7.27 (m, 3H), 7.25 (d, J = 1.8 Hz, 1H), 7.17 - 7.10 (m, 2H), 6.86 - 6.79 (m, 2H), 4.70 (t, J = 5.2 Hz, 1H), 3.89 - 3.80 (m, 2H), 3.58 (dq, J = 9.5, 7.0 Hz, 2H), 3.47 (dq, J = 9.5, 7.0 Hz, 2H), 1.05 (td, J = 7.0, 0.9 Hz, 6H). 13 C NMR (126 MHz, d6-DMSO) δ 161.88, 159.18, 151.76, 144.86, 143.32, 141.77, 130.92 (q, J = 32.3 Hz), 129.83, 129.68, 126.75, 125.13 (d, J = 2.7 Hz), 123.60, 122.56, 119.34, 118.79, 118.44, 115.21, 107.11, 100.21, 74.12, 68.38, 62.21, 15.63.HRMS(ESI)m / z:[M+H]+C 29 H 27 Calculated value of F3NO5: 526.1763, measured value: 526.1777.

[0166] Alternative preparation 17 (R)-5-4-(2,2-diethoxyethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol Combine (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol (2 g) and tert-amyl alcohol. Add sodium tert-pentoxide (0.85 g) and maintain the temperature at 20-30° C. for 19 hours. Add additional sodium tert-pentoxide (0.2 g) and stir at 20-30° C. for 4 hours to give the crude title compound.

[0167] Alternative preparation 17a (R)-5-4-(2,2-diethoxyethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol Nitrogen is bubbled through a solution of (R)-4-((4-(2,2-diethoxyethoxy)phenyl)(hydroxy)methyl)-3-(2-fluoro-4-(trifluoromethyl)phenyl)quinolin-7-ol (18.4 g, 34.8 mmol, 96.3% ee) and sodium tert-pentoxide (11.3 g, 102.6 mmol) in 2-MeTHF (100 mL) at 25 °C for approximately 30 min to 1 h. Stir at 25 °C for 16 h. Wash with NaHSO (1 M, 60 mL), followed by 7% NaHCO (60 mL), and then water (60 mL). Concentrate the solution to approximately 60 mL, and crystallize the material by adding heptane. The precipitated solid is filtered, washed with 2-MeTHF / heptane solution, and dried under vacuum to give the title compound (14.1 g, 77% yield, 95.6% ee). The chiral analysis conditions for the title compound and starting material are from Preparation 17 and Preparation 13, respectively.

[0168] Preparation 18 [ka] Scheme 4, Step E: As shown above, it should be noted that the aldehyde form may be in equilibrium with a hydrated aldehyde form.

[0169] Under a nitrogen atmosphere, (R)-5-4-(2,2-diethoxyethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol (248.1 g, 80.6% purity = 200.0 g; 92.4% ee; 381.0 mmol; purity analysis conditions are as described in Preparation 17) is combined with ACN (1.0 L) and aqueous 1 N HCl (350 mL). The resulting mixture is stirred at 60 °C for 2 h, cooled to 25 °C, and circulated through activated carbon (1.5 g) for 1.0 h. 7% aqueous NaHCO (350 mL) is added over 1.0 h to adjust the pH to 3 while maintaining the temperature at 20-30 °C. The resulting mixture is concentrated under vacuum to 1.4 L at 35-45 °C. Water (2 L) is added over 3.0 h and the pH is adjusted to 5.9 with 7% NaHCO3 (140 mL) at 25 °C. The mixture is stirred at 25 °C for 2 h and then at 5 °C for 48 h. The resulting solid is filtered, washed with water (400 mL), the solid is suspended, and ACN (600 mL) is added at 45 °C for 2.0 h and then stirred at 5 °C for 2.0 h. The resulting solid is filtered, washed with ACN (400 mL), and dried under vacuum at 50 °C for 16 h to give the title compound as a yellow solid (157.8 g, 99.6% ee, 87.2% purity, 80.1% yield). Purity analysis conditions: Shimadzu LC-20A HPLC system, Waters Cortecs C18+ column (100mm*4.6mm, 2.7mM), column temperature 45℃, gradient of 80% A (0.1% TFA in H2O) / 20% B (0.1% TFA in AcCN) to 30% A / 70% B over 17 min, then 5% A / 95% B over 2 min, flow rate 1.0mL / min, UV 270nm. Chiral analysis conditions: Agilent 1290 UPLC system, Chiralpak IG (250mm*4.6mm, 5μm), column temperature 45℃, gradient of 25 to 55% ACN (0.1% TFA) in water (0.1% TFA) over 30 min, flow rate 0.3mL / min, UV 270nm. (R) Elutes at 24.2 min. Aldehyde form: (approximately 50% molar in d6-DMSO solution): 11H NMR (500 MHz, d6-DMSO) δ 10.54 (s, 1H), 9.61 (s, 1H), 9.52 (s, 1H), 8.35 (d, J = 7.7 Hz, 1H), 7.84 (dd, J = 9.19, 7.00 Hz, 1H), 7.39 (m, 1H), 7.38 (m, 1H), 7.32 (d, J = 5.62 Hz, 1H), 6.48 (dd, J = 6.48, 1.81 Hz, 1H), 7.21 (dt, J = 9.1, 2.5 Hz, 1H), 7.11 (dd, J = 8.9, 7.3 Hz, 2H), 6.81 (dd, J = 8.8, 6.4 Hz, 2H), 4.78 (s, 2H). 13 13C NMR (126 MHz, d6-DMSO) δ 198.76, 159.38, 157.97, 151.41, 149.09, 145.83, 136.80, 130.22, 129.80 (J = 31.8 Hz), 129.19, 125.53, 124.24, 123.98, 123.7 (q, J = 272.8 Hz), 120.52, 118.73 (m), 117.51, 117.13, 114.71, 114.6 (m), 110.70, 73.61, 72.40. HRMS(ESI) m / z: [M+H] + C 25 H 17 Calculated value for C19H13F3NO4: 452.1031, found: 452.1025.

[0170] Hydrated aldehyde form from d6-DMSO (ca. 50 mol% in d6-DMSO solution): 1H NMR (500 MHz, d6-DMSO) δ 10.54 (s, 1H), 9.52 (s, 1H), 8.35 (d, J= 7.7 Hz, 1H), 7.84 (dd, J= 9.19, 7.00 Hz, 1H), 7.39 (m, 1H), 7.38 (m, 1H), 7.32 (d, J= 5.62 Hz, 1H), 6.48 (dd, J= 6.48, 1.81 Hz, 1H), 7.21 (dt, J = 9.1, 2.5 Hz, 1H), 7.11 (dd, J = 8.9, 7.3 Hz, 2H), 6.81 (dd, J = 8.8, 6.4 Hz, 2H), 6.03 (br, 1H), 5.00 (t, J = 5.1 Hz, 1H), 3.70 (dd, J = 5.1, 1.1 Hz, 2H). 13 C NMR (126 MHz, d6-DMSO) δ 159.38, 158.85, 151.43, 149.09, 145.83, 136.88, 130.22, 129.80 (J = 31.8 Hz), 129.19, 125.53, 124.24, 123.98, 123.7 (q, J = 272.8 Hz), 120.52, 118.73 (m), 117.51, 117.13, 114.71, 114.6 (m), 110.70, 87.97, 73.73, 71.58.HRMS(ESI)m / z:[M+H] + C25H 18 Calculated value of F3NO5: 469.40932, measured value: 470.1288.

[0171] Alternative preparation 18 (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde Combine (R)-5-4-(2,2-diethoxyethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol (0.99 g, 1.9 mmol) and formic acid (4 mL). Stir overnight at room temperature. Add a solution of 28% by weight ammonium hydroxide (2 mL, 14 mmol) in water (15 mL) over 5 minutes. Filter the solution, wash with ACN (2 mL), and dry in a vacuum oven at 40 °C to give the title compound (0.79 g, 92% yield).

[0172] Alternative preparation 18a (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde Combine (R)-5-4-(2,2-diethoxyethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol (5 g, 9.5 mmol), ACN (15 mL), water (10 mL), and TsOH (2.2 g, 11.4 mol). Heat the solution at 70 °C for 5 h and cool to room temperature. Adjust the pH to 5.0-5.5 with 7% NaHCO3 (aqueous, 3.4 mL) and stir at approximately 25 °C for 30 min. A precipitate forms in approximately 10 min. Filter the resulting precipitate, wash with water, and dry under vacuum to obtain the title compound (4.29 g, 98.2% purity, 99.7 ee, 87% yield). The purity analysis conditions for this product are described in Preparation 18.

[0173] Preparation 18b [ka] The title compound is prepared as described in US Pat. No. 10,654,866 for the racemate and the compound is purified as described for Isomer 1 to give the title compound.

[0174] Preparation 19 [ka] Scheme 4, Step F: Under a nitrogen atmosphere, 3-(fluoromethyl)azetidine tosylate (37.6 g, 143.9 mmol) is combined with anhydrous EtOH (250 mL) and TEA (11.2 g, 110.7 mmol). The resulting mixture is stirred at room temperature to obtain a clear solution. In a separate reactor, a mixture of (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde (57.5 g, 87% purity, 110.8 mmol; purity analysis conditions described in Preparation 18) and anhydrous EtOH (250 mL) is cooled to 0-10 °C and STAB (47.0 g, 221.5 mmol) is added. Add the 3-(fluoromethyl)azetidine mixture, TEA, and EtOH solution to the (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde mixture over 1 h at 0-10 °C, and stir the mixture at 5-15 °C for 1 h. With continued stirring, add 10% aqueous NH4Cl (250 mL) to the reaction mixture, followed by concentration to a final volume of approximately 375 mL. Add DCM (500 mL) to the resulting mixture, and separate the organic phase. The organic layer is washed with 10% aqueous Na2CO3 (250 mL) and 10% aqueous NaCl (250 mL) and circulated through activated carbon (1.5 g) for 30 min to give the title compound as a solution in DCM (500 mL) (58.7 g in DCM solution, 99.5% ee, 100% yield. Chiral analysis conditions: Shimadzu LC-20A HPLC system, Chiralpak IC (250 mm*4.6 mm, 5 μm), column temperature 40 °C, 90% hexane (0.1% DEA) / 10% EtOH over 30 min, flow rate 1.0 mL / min, UV 270 nm, t (R) Elutes at 14.2 minutes.

[0175] solid state NMR (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol Solid-state NMR was acquired on a Bruker Avance III HD equipped with a Bruker Ultrashield 400WB Plus magnet operating at a frequency of 100.6 MHz. The probe used was a Bruker MAS 4 BL CP BB DVT NP / H. Acquisition parameters were as follows: 7776 scans, 34 ms acquisition time, 8.5 s interpulse delay, 10 kHz MAS frequency, 1.5 ms contact time, and SPINAL64 decoupling scheme. Data were externally referenced to 29.5 ppm adamantane. 13 C solid-state NMR (101 MHz)δ 161.2, 159.0, 153.2, 151.3, 146.5, 135.1, 132.0, 130.8, 130.0, 125.2, 123.7, 122.6, 118.8, 117.9, 117.2, 114.7, 111.3, 84.9, 83.2, 74.8, 62.8, 57.8, 56.0, 53.3, 28.0.

[0176] Alternative preparation 19 (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol The title compound is prepared as described in US Pat. No. 10,654,866 and the enantiomers are separated to give Isomer 2, the title compound of Preparation 19.

[0177] Alternative preparation 19a [ka] A 100 mL flask was charged with (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-5-yl)phenoxy)ethane-1,1-diol (10 g, 20.2 mmol). This intermediate was assumed to be 95% potent and contained 5% water by weight. A stir bar and THF (50 mL) were added to the flask, the flask was fitted with a septum, and the reaction was blanketed with nitrogen gas. Water (135 mL) was added to adjust the water content to 10% by weight. The reaction was stirred at room temperature for 2 hours to give a clear, orange-brown solution.

[0178] A separate 250 mL jacketed reactor equipped with mechanical stirring and a positive nitrogen atmosphere was charged with 3-(fluoromethyl)azetidine tosylate (5.6 g, 21.3 mmol), THF (48 mL), and triethylamine (1.4 mL). The jacket temperature was set to 0 °C, and the reaction was stirred at 0 °C for 30 minutes. Sodium triacetoxyborohydride (8.15 g, 38.5 mmol) was added in one portion. The prepared THF solution of (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-5-yl)phenoxy)ethane-1,1-diol was added via syringe pump and added dropwise to the jacketed reactor over 2 hours, maintaining the reaction temperature at 0 °C. After the addition was complete, the reaction was stirred at 0 °C for an additional 2 hours. The reaction was analyzed for completion. The reaction was complete when the amount of 1,1-diol was less than 1% relative to (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol. If the amount of 1,1-diol was greater than 1%, the reaction was stirred at 0°C for an additional 2-8 hours until complete.

[0179] The completed reaction was treated with 15% aqueous KHCO3 (57 mL) over 10 minutes at 0°C. The reaction jacket temperature was raised from 20°C to 30°C and the reaction was stirred for 2 hours. The stirring was stopped and the reaction was allowed to stand for 1 hour. The aqueous layer was drained and the reaction was recharged with 15% aqueous KHCO3 (57 mL). The jacket temperature was raised from 40°C to 50°C and the reaction was stirred at that temperature for 2 hours. The stirring was stopped and the reaction was allowed to stand at 20°C to 30°C. The reaction was analyzed for completion. If the tosylate peak was greater than 1% relative to (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol, washing with 15% aqueous KHCO was continued as described until the tosylate peak was less than 1%.

[0180] The aqueous layer was drained and the reaction was treated with heptane (4.5 mL) and water (38 mL). The reaction was stirred at 20-30°C for 30 min, stirring was stopped, the reaction was allowed to stand for 1 h, and the aqueous layer was drained.

[0181] The reaction was concentrated under slight vacuum (target -0.05 MPa) with a bath temperature below 70 °C to a total volume of 60 mL - 100 mL. THF (95 mL) was added and the reaction was concentrated under slight vacuum (target -0.05 MPa) with a bath temperature below 70 °C to a total volume of 30 mL - 50 mL. If the reaction contained more than 1% water by KF analysis, the above distillation step was repeated.

[0182] The reaction volume was adjusted to 60 mL with THF and stirred at 40°C for 1 hour. MeCN was added via syringe pump over 1 hour at 40°C, then the reaction was seeded with (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol (70 mg) and the reaction was stirred at 40°C for 4 hours. MeCN (38 mL) was added via syringe pump over 2 hours and stirred at 40°C for an additional 2 hours. MeCN (57 mL) was added via syringe pump over 3 hours and stirred at 40°C for an additional 1 hour. The temperature was adjusted to 0°C-10°C over 3 hours and stirred at 0°C for an additional 2 hours. The solids were removed by filtration and rinsed with MeCN / THF (29 mL of a 2:1 solution). The solid was dried under vacuum at 30° C. for 5 hours and at 45° C. for 11 hours to give the title compound (7.7 g, 70%).

[0183] Preparation 20 [ka] A 60% dispersion of NaH in mineral oil (0.10 g, 2.5 mmol) is added to a solution of 3-[2-fluoro-4-(trifluoromethyl)phenyl]-4-[hydroxy-[2,3,5,6-tetradeuterio-4-[2-[3-(fluoromethyl)azetidin-1-yl]ethoxy]phenyl]methyl]quinolin-7-ol (0.17 g, 0.31 mmol) in THF (12 mL) and heated at 50–60 °C overnight. The reaction is cooled to room temperature and diluted sequentially with saturated ammonium chloride (10 mL) and saturated sodium bicarbonate (20 mL). The mixture is extracted with DCM (2 × 50 mL), and the combined organic layers are washed with saturated brine (10 mL), dried over sodium sulfate, and concentrated onto silica gel. Purification on silica gel, eluting with a gradient of 3–8% MeOH in DCM, affords the title compound (0.09 g, 55%) as a yellow solid. Two essentially identically prepared batches were combined in MeOH and DCM and dried under vacuum overnight to give the final material. 1H NMR (400 MHz, DMSO-d6) δ 10.41 (s, 1H), 9.49 (s, 1H), 8.46 - 8.26 (m, 1H), 7.81 (d, J = 9.2 Hz, 1H), 7.39 (ddd, J = 8.1, 1.9, 0.8 Hz, 1H), 7.34 (d, J = 2.5 Hz, 1H), 7.30 (s, 1H), 7.23 (d, J = 1.9 Hz, 1H), 7.17 (dd, J = 9.1, 2.5 Hz, 1H), 4.51 (d, J = 6.2 Hz, 1H), 4.40 (d, J = 6.3 Hz, 1H), 3.84 - 3.75 (m, 2H), 3.25 (td, J = 7.6, 1.5 Hz, 2H), 2.97 - 2.88 (m, 2H), 2.75 - 2.57 (m, 3H). 19 F NMR (376 MHz, DMSO-d6) δ 17.40 (td, J = 47.6, 46.8, 18.1 Hz), -61.16 (s). 13 C NMR (101 MHz, DMSO) δ 159.62, 151.93, 150.03, 146.57, 136.91, 129.85, 125.94, 124.85, 124.39, 120.78, 117.89, 117.49, 111.54, 85.90, 84.28, 74.12, 66.70, 57.61, 56.50, 56.42, 40.66, 40.45, 40.24, 40.03, 39.82, 39.61, 39.40, 31.25, 31.06.

[0184] Example 1

change

[0185] Alternative Example 1 (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol, 4-methylbenzenesulfonic acid Scheme 4, Step F: Add 3-(fluoromethyl)azetidine tosylate (0.79 g, 0.300 mmol) and IPA (5 mL) and stir the mixture. Add pyridine borane (200 μL, 2.0 mmol) to the mixture. In a separate container, add (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde (1.00 g, 2.21 mmol) in DMSO (2.5 mL) and add dropwise to the tosylate mixture over approximately 5 minutes. Stir for 3 hours. Add IPA (15 mL) dropwise. Filter the resulting solid, wash the wet cake with IPA (2 × 5 mL), and dry the solid in a vacuum oven to give the title compound (1.09 g, 68% yield). Essentially the same procedure is completed using DMAC as the solvent to give the title compound (1.06 g, 66% yield).

[0186] Alternative Example 1a (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol, 4-methylbenzenesulfonic acid Scheme 4, Step F: Combine pyridinium p-tosylate (0.5021 g, 2.0 mmol), sodium borohydride (76.0 mg, 2.01 mmol), and THF (10 mL). Stir for 3 hours and filter through diatomaceous earth. Add 3-(fluoromethyl)azetidine tosylate (766.1 mg, 2.9 mmol) in tert-amyl alcohol (15 mL) and stir the mixture for 1 hour. Add (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde (1.01 g, 2.2 mmol) in THF (5 mL) via syringe pump over 2 hours and stir the mixture overnight. The mixture is filtered, and the wet cake is washed with IPA (1.5 mL) and dried in a vacuum oven at 40° C. to give the title compound (1.16 g, 68% yield). 1 H NMR (600 MHz, ACN-d3 / D2O, 25 ℃) δ 9.31 (s, 1H), 8.10 (d, J = 8.1 Hz, 1H), 7.65 (d, J = 9.1 Hz, 1H), 7.61 - 7.57 (m, 2H), 7.39 (d, J = 2.5 Hz, 1H), 7.33 (dd, J = 8.2, 1.8 Hz, 1H), 7.19 - 7.12 (m, 4H), 7.12 - 7.08 (m, 2H), 7.07 (s, 1H), 6.77 - 6.72 (m, 2H), 4.48 (dd, J = 47.0, 3.8 Hz, 2H), 4.26 - 4.19 (m, 2H), 4.05 - 3.99 (m, 4H), 3.44 (m, 2H), 3.22 - 3.04 (m, 1H), 2.29 (s, 3H).

[0187] Alternative Example 1b (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol, 4-methylbenzenesulfonic acid Scheme 4, Step F: Combine (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde (2.0 g, 4.3 mmol) and THF (20 mL). Stir for 30 minutes. In a separate vessel, combine sodium triacetoxyborohydride (2.0 g, 9.5 mmol), THF (20 mL), and 3-(fluoromethyl)azetidine tosylate (1.2 g, 2.3 mmol). Add the THF solution of (R)-2-(4-(2-hydroxy-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-5-yl)phenoxy)acetaldehyde dropwise over 1 hour. Stir for an additional 1 hour. Quench with 10% aqueous KHSO4 (10 mL) and stir for 30 minutes. Filter and wash the organic layer with 10% aqueous NaHCO3 (10 mL) and saturated NaCl. Concentrate the organic layer, crystallize the residue from THF / ACN, filter the precipitated solid, and dry the solid in a vacuum oven to give the title compound (1.02 g, 41% yield).

[0188] Solid State NMR, Example 1b Solid-state NMR was obtained with an interpulse delay of 7.5 seconds, similar to preparation 19. 13 C solid-state NMR (101 MHz)δ 162.0, 158.6, 152.2, 148.9, 143.6, 143.1, 141.7, 139.0, 138.5, 131.1, 129.1, 128.1, 126.9, 125.3, 123.3, 121.8, 121.0, 118.6, 117.9, 116.9, 116.2, 110.1, 108.4, 84.2, 82.6, 81.1, 72.2, 61.4, 57.4, 55.6, 54.8, 30.9, 29.0, 20.9.

[0189] Example 2 [ka] (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol (293 mg, 0.56 mmol) and water (0.3 mL) were combined in ACN (6 mL) and heated to 50° C. with stirring to give a yellow solution. Benzenesulfonic acid (0.108 mg, 0.68 mmol) was dissolved in ACN (1 mL) and added to the (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol solution to give a light yellow solution. Stir at 50° C. for 2 hours, then cool to room temperature. The resulting precipitate is filtered under vacuum and air-dried without vacuum over the weekend to give the title compound (0.157 g, 41%). Solid-state NMR is obtained as in preparation 19 with an interpulse delay of 8.0 seconds. 13 C solid-state NMR (101 MHz) δ 161.9, 159.1, 158.4, 151.8, 149.1, 146.8, 146.4, 142.8, 138.0, 131.5, 130.4, 129.7, 128.2, 127.5, 126.6, 125.1, 122.9, 122.3, 121.5, 120.8, 118.5, 117.8, 117.0, 116.3, 110.4, 109.2, 108.5, 84.2, 83.0, 82.5, 81.4, 71.9, 65.0, 64.3, 62.0, 61.7, 61.0, 60.4, 60.0, 58.2, 57.9, 57.2, 56.8, 56.4, 55.2, 54.5, 54.0, 53.6, 52.6, 31.6, 31.0, 30.3, 29.6.

[0190] Example 3 [ka] The final bond formation step, i.e., reductive amination, in the synthesis of (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol, 4-methylbenzenesulfonic acid is important for controlling the purity of the final compound. The most important aspect is that the use of triethylamine in the reductive amination limits the formation of (5R)-5-[4-(2-hydroxyethoxy)phenyl]-8-(trifluoromethyl)-5H-chromeno[4,3-c]quinolin-2-ol to 0.3% or less, as measured by HPLC. 1 H NMR (500 MHz, DMF-d7Rt, 25°C) δ 10.72 (s, 1H), 9.61 (s, 1H), 8.49 - 8.44 (m, 1H), 7.95 (d, J = 9.1 Hz, 1H), 7.50 (d, J = 2.5 Hz, 1H), 7.46 (m, 1H), 7.38 (s, 1H), 7.32 - 7.26 (m, 2H), 7.23 (d, J = 8.8 Hz, 2H), 6.87 (d, J = 8.8 Hz, 2H), 4.89 (t, J = 5.7 Hz, 1H), 3.99 (dd, J = 5.4, 4.6 Hz, 2H), 3.78 (td, J = 5.7, 4.5 Hz, 2H).MS ES+ m / z 454 [M+H] + Purity analysis conditions: Agilent 1260 HPLC system, Waters Cortecs C18+ column (100 mm x 4.6 mm, 2.7 mm), column temperature 45 °C, elution with a gradient of 75% A (0.1% TFA in water:ACN:MeOH = 90:2:8, v / v / v) / 25% B (ACN:MeOH = 20:80, v / v) to 10% A / 90% B over 30 min, flow rate 0.8 mL / min, UV 270 nm. Rt = 14.661 min.

[0191] Example 4 [ka] 3-(Chloromethyl)azetidine 4-methylbenzenesulfonate is an impurity that may be present in 3-(fluoromethyl)azetidine 4-methylbenzenesulfonate. When present, it typically amounts to between 0.05% and 0.5% as determined using HPLC. 1 H NMR (500 MHz, DMSO) δ 8.71 (s, 2H), 7.57 (d, J = 8.1 Hz, 2H, counterion), 7.16 (d, J = 8.1 Hz, 2H, counterion), 4.05 (dd, 2H), 3.84 (d, J = 6.8 Hz, 2H), 3.81 (dd, 2H), 3.18 (m, 1H), 2.33 (s, 3H, counterion).

[0192] Example 5 [ka] The title compound above may be formed when 3-(chloromethyl)azetidine 4-methylbenzenesulfonic acid is present during the synthesis of imlunestrant. This compound may be present at levels of 0.05% to 0.5%. MS ES+ m / z 541 [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ 10.50 (s, 1H), 9.50 (s, 1H), 8.34 (d, J = 8.1 Hz, 1H), 7.81 (d, J = 9.2 Hz, 1H), 7.37 (dd, J = 8.1, 1.6 Hz, 1H), 7.37 (d, J = 2.5 Hz, 1H), 7.28 (s, 1H), 7.21 (d, J = 1.6 Hz, 1H), 7.18 (dd, J = 9.2, 2.5 Hz, 1H), 7.09 (d, 8.8 Hz, 2H), 6.76 (d, 8.8 Hz, 2H), 3.79 (m, 2H), 3.71 (d, J = 7.5 Hz, 2H), 3.26 (dd, J = 7.3 Hz, 2H), 2.88 (dd, J = 7.4, 6.2 Hz, 2H), 2.63 (dd, J = 7,4, 5.7 Hz, 2H), 2.63 (m, 1H). Purity analysis conditions: Agilent 1260 HPLC system, Waters Cortecs C18+ column (100 mm x 4.6 mm, 2.7 mm), column temperature 45 °C, gradient from 75% A (0.1% TFA in water:ACN:MeOH = 90:2:8, v / v / v) / 25% B (ACN:MeOH = 20:80, v / v) to 10% A / 90% B over 30 min, flow rate 0.8 mL / min, UV 270 nm.

[0193] Example 6 [ka] The above compound was present as an impurity found in a sample of (R)-5-(4-(2-(3-(fluoromethyl)azetidin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-c]quinolin-2-ol, 4-methylbenzenesulfonic acid. This compound is believed to form when any free chloride ion present reacts with the cyclobutyl group. MS ES+ m / z 561 [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ 10.43 (s, 1H), 9.50 (s, 1H), 8.36 (d, J = 8.2 Hz, 1H), 7.82 (d, J = 9.2 Hz, 1H), 7.40 (dd, J = 8.2, 2.1 Hz, 1H), 7.36 (d, J = 2.5 Hz, 1H), 7.31 (s, 1H), 7.24 (d, J = 2.1 Hz, 1H), 7.18 (dd, J = 9.2, 2.5 Hz, 1H), 7.10 (d, 8.8 Hz, 2H), 6.82 (d, 8.8 Hz, 2H), 4.49 (m, 2H), 3.91 (m, 2H), 3.70 (m, 2H), 2.82 (dd, J = 11.2 Hz, 2H), 2.61 (d, J = 6.7 Hz, 2H), 2.18 (m, 1H)

[0194] Example 7 [ka]

[0195] Example 8 [ka] MS ES+ m / z 1050 [M+H] + .

[0196] Chromatography Methods Two different chromatographic protocols are used to compare the compounds of Preparation 18b, Preparation 19, and Alternative Preparation 19: HPLC Method 1 and HPLC Method 2, which are described below.

[0197] HPLC Method 1 utilizes the following conditions: an Agilent 1260 HPLC system equipped with a 245 nm UV detector, an Agilent Bonus RP 4.6 x 75 mm, 3.5 μm column, a column temperature of 30°C, a flow rate of 1.0 mL / min, and an injector volume of 5 μl. Mobile phase A consists of 0.05% TFA in water, and mobile phase B consists of 0.05% TFA in ACN. Samples are eluted with a gradient of 10% B to 50% B in 20 minutes and 95% B in 25 minutes. In System 1, a 0.8 mg / mL standard solution is prepared by dissolving 40 mg of sample and diluting to 50 mL with a 60% ACN / 40% water diluent.

[0198] HPLC Method 2 utilizes the following conditions: an Agilent 1260 HPLC system equipped with a 270 nm UV detector, a Waters Cortecs C18+, 4.6 x 100 mm, 2.7 μm column, a column temperature of 45°C, a flow rate of 0.8 mL / min, and an injector volume of 2 μl. Mobile phase A consists of 0.1% TFA in water:ACN:MeOH = 90:2:8, v / v / v. In one example, 1800 mL of water is mixed with 200 mL of mobile phase B. 2.0 mL of TFA is accurately transferred and mixed thoroughly. Mobile phase B consists of 20:80 ACN:MeOH, v / v. In one example, 400 mL of ACN and 1600 mL of MeOH are mixed thoroughly and degassed by ultrasound. The sample is eluted with a gradient from 75% A and 25% B to 10% A and 90% B over 30 minutes. For System 2, prepare the compound in a 0.5 mg / mL solution by dissolving 50 mg of sample and dilute to 100 mL with a 1 / 1 solution of ACN / water.

[0199] HPLC Method 1 and HPLC Method 2 can be used to determine the percent area (% area) of the compound of Formula A, Formula B, or Formula C, as well as dihydroquinoline or quinoline-based impurities. As used herein, "% area" refers to the percent area obtained using HPLC Method 1 or HPLC Method 2, or both. For the avoidance of doubt, the % area of ​​the compound of Formula A plus the % area of ​​any contaminants or impurities will always be less than or equal to 100% area, the % area of ​​the compound of Formula B plus the % area of ​​any contaminants or impurities will always be less than or equal to 100% area, and the % area of ​​the compound of Formula C plus the % area of ​​any contaminants or impurities will always be less than or equal to 100% area.

[0200] Figures 1 and 2 show a peak at approximately 8.8 minutes for Preparation 18b and Alternative Preparation 19 using Chromatography System 1. This peak is the major product of the Preparation 18b reaction. The peak at approximately 8.8 minutes is not seen in the Preparation 19 sample. The % area of ​​the peak at 8.8 minutes in Alternative Preparation 19 is 0.23%.

[0201] Figure 3 shows a peak at approximately 5.2 minutes in Preparation 18b that is barely visible in Preparation 19 utilizing chromatographic system 2. The area of ​​the peak at approximately 5.2 minutes is not seen in the Preparation 19 sample. The % area of ​​the peak at 5.2 minutes in alternative Preparation 19 is <0.05% area, below the limit of quantitation for the method described herein. The difference in % area between the different systems can be explained by the difference in the response factor of the compound for UV 245 nm in System 1 versus UV 270 nm in System 2.

[0202] Preparation 19 was used to synthesize several batches of (R)-5-(4-(2-(3-(fluoromethoquinolinedin-1-yl)ethoxy)phenyl)-8-(trifluoromethyl)-5H-[1]benzopyrano[4,3-quinolin-2-ol] with the following purities (% area): [Table 8]

[0203] In one embodiment, the compound of Formula B, or a pharmaceutically acceptable salt thereof, is at least 98.5% area, at least 98.6% area, at least 98.7% area, at least 98.8% area, at least 98.9% area, at least 99.0% area, at least 99.1% area, at least 99.2% area, at least 99.3% area, at least 99.4% area, at least 99.5% area, at least 99.6% area, at least 99.7% area, or at least 99.8% area. In one embodiment, the pharmaceutically acceptable salt is a tosylate salt.

[0204] Biological assays High-content imaging assay for ERα degradation in MCF7 cells Cells were fixed by adding 10 μL of 14% paraformaldehyde for 30 minutes at room temperature. Cells were washed once with 20 μL of PBS and incubated with 20 μL of PBS containing 0.5% (v / v) TWEEN® 20 per well for 1 hour. Cells were washed twice with 0.05% TWEEN® 20 in PBS and blocked with 20 μL of 3% BSA in PBS containing 0.05% TWEEN® 20 and 0.1% TRITON® X-100 for 1 hour at room temperature. A 1:500 dilution of primary antibody (20 μL) (ERα (clone SP1) monoclonal rabbit antibody #RM-9101-S, Thermo Scientific) in 1% BSA in PBS containing 0.05% TWEEN® 20 was added, the plate was sealed, and the plate was incubated overnight at 4°C. The next day, cells were washed twice with PBS containing 0.05% TWEEN® 20 and incubated with secondary antibody (20 μL / well) (1:1000 dilution, goat anti-rabbit IgM ALEXA FLUOR™ 488) in PBS 1% BSA for 105 minutes at room temperature. After incubation, plates were washed with PBS (2 × 20 μL), and PBS (20 μL) containing RNase (Sigma) (50 μg / mL) and a 1:1000 dilution of propidium iodide was added per well. Plates were sealed and incubated for 1 hour at room temperature (protected from light). Plates were scanned using an ACUMEN EXPLORER™ (a laser-scanning fluorescence microplate cytometer manufactured by TTP LABTECH LTD) to measure ERα. Image analysis was based on cytofluorescence signals to identify positive cells. Estrogen receptor-positive cells were identified by average intensity. The total intensity from propidium iodide / DNA at 575-640 nm was used to identify individual cells. The assay output was the % estrogen receptor-positive cells. IC was calculated by curve fitting a four-parameter logistic regression of each output using GENE DATA™. 50was determined. The raw data (FLU) was plotted as a nonlinear regression via the GENEDATA SCREENER® tool. Data were analyzed using a four-parameter nonlinear logistic equation (four-parameter logistic concentration-response curve): Y = bottom + [(top - bottom) / 1 + (x / IC 50 ) slope] where Y = % inhibition, x = concentration resulting in y % inhibition, bottom = minimum value of y obtained by the curve, top = maximum value of y obtained by the curve, and slope = IC 50 The sharpness of the curve at %Inh = [(median maximum - x / median maximum - median minimum)] x 100. The results are shown in Table 2, SERD Contaminants, and demonstrate that synthetic compounds comparable to Formula A, intermediates in the process of Formula A, are not active SERD compounds at the concentrations tested. In Table 2, compounds with a relative IC50 > 2.00 μM are inactive. [Table 9-1] [Table 9-2]

[0205] Other embodiments From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adapt it to various applications and conditions, and such embodiments also fall within the scope of the following claims.

[0206] The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of the listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0207] All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each individual patent and publication was specifically and individually indicated to be incorporated by reference.

Claims

1. Compound of Formula A 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, wherein R １ or R ２ are independently Cl, F, or —CF ３ , or -CH ３ and the other is H, and R ７ is H or PG, and said PG is C １ ~C ４ Alkyl, benzyl, benzoyl, C １ ~C ６ alkanoyl, methoxyethoxymethyl ether group, silyl ether group, tetrahydropyranyl, tetrahydrofuranyl, and ethoxyethyl ether, wherein the compound of Formula A has a purity of at least 98% area and contains less than 1% area of ​​one or more dihydroquinoline or quinoline-based impurities as measured using HPLC, and the dihydroquinoline or quinoline-based impurities are selected from the following compounds: Table 1 【change】 【change】 【change】 【change】 【change】 A composition selected from:

2. The dihydroquinoline or quinoline-based impurity is selected from the group consisting of the following compounds: Table 2 The composition of claim 1 selected from:

3. The dihydroquinoline or quinoline-based impurity is selected from the group consisting of the following compounds: Table 3 The composition of claim 1 selected from:

4. A compound of formula B having a purity of at least 98.5% area as measured using HPLC. 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

5. The composition of claim 1 , wherein the pharmaceutically acceptable salt is a tosylate salt.

6. A pharmaceutical composition comprising the composition of any one of claims 1 to 5 in combination with at least one pharmaceutically acceptable excipient, carrier, or diluent.

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