Process of preparing repotrectinib

Abstract

Disclosed are process for preparing repotrectinib, compound of Formula (I) and a process for preparing compound (D). Formula (I) and Formula (D).

Description

[0001] PROCESS OF PREPARING REPOTRECTINIB

[0002] CROSS REFERENCE TO RELATED APPLICATION

[0003] This application claims the benefit of U. S. Provisional Application No. 63 / 632,580, filed April 11, 2024, the content of which is hereby incorporated herein by reference in its entirety.

[0004] REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY VIA PATENT CENTER

[0005] Incorporated herein by reference in its entirety is a Sequence Listing entitled, “20250408 SEQ 14476WOPCT” comprising of SEQ ID NO: 1, which include nucleic acid and / or amino acid sequences disclosed herein. The Sequence Listing has been submitted herein in XML format via Patent Center, and thus constitutes both the paper and computer readable form thereof. The Sequence Listing was first created using WIPO Sequence on April 8, 2025, and is 2.17 KB.

[0006] FIELD OF THE INVENTION

[0007] The present invention relates to methods for preparing the compound (75,13 / ?)- 11 -fluoro-7, 13 -dimethyl-6, 7, 13,14-tetrahy dro- 1,15- ethenopyrazolo[4, 3 - y][l,4,8,10]benzoxatriaza-cyclotridecin-4(5J7)-one and intermediates thereof.

[0008] BACKGROUND OF THE INVENTION

[0009] The compound repotrectinib, also known as (75,137?)-1 l-fhioro-7,13-dimethyl- 6,7,13,14-tetrahydro-l,15- ethenopyrazolo[4,3- / |[l,4,8,10]benzoxatriaza-cyclotridecin- 4(5J7)-one, the compound is also known as (3R,l lS)-6-Fluoro-3,l l-dimethyl-10-oxa- 2,13, 17,18,21-pentaazatetracyclo[13.5.2.04’9.018’22]docosa-l(21),4,6,8,15(22),16,19- heptaen- 14-one (also herein referred to as "Compound I") i s represented by F ormula ( I)

[0010] I.

[0011] Compound (I) is a potent small-molecule multi-target kinase inhibitor showing activity against wild- type and mutant ALK (anaplastic lymphoma kinase), wild-type and mutant ROSI (ROSI proto- oncogene receptor tyrosine kinase), the TRK family of kinases (tropomyosin-related receptor tyrosine kinases), JAK2 of the Janus family of kinases, SRC (Src family of protein tyrosine kinases (SFKs)) and FAK (focal adhesion kinase). Compound (I) has properties, including anti-tumor properties, that are pharmacologically mediated through inhibition of tyrosine kinase receptors. Compound (I) was disclosed in International Patent WO2015 / 112806 and in W02017 / 007759, which are incorporated herein by reference in its entirety.

[0012] Compound (I) has found application in treating disease associated with receptor tyrosine kinases, such as ALK, ROSI, TRK, JAK2, SRC and FAK. It is advantageous to have improved methods of preparation.

[0013] SUMMARY OF THE INVENTION

[0014] The present invention provides methods of preparing the compound of formula (I), along with intermediates thereof.

[0015] The present invention also provides processes and intermediates for making the compounds of the present invention.

[0016] Additionally, the present invention provides a process for making compound (D).

[0017] These and other features of the invention will be set forth in expanded form as the disclosure continues.

[0018] DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

[0019] In one aspect, the present invention provides, a process for preparing a compound of formula (I): comprising the steps of:

[0020] 1) reacting compound (A) and compound (B) in the presence of a base to give compound (C)

[0021] 2) with or without isolating compound (C), contacting compound (C) with an asymmetric transamination (ATA) enzyme to give compound (D) optionally, converting compound (D) into a salt;

[0022] 3) contacting compound (D), either as a free base or as a salt, with compound (E) to give compound (F);

[0023] 4) with or without isolating compound (F), treating compound (F) with a base to give compound (G), compound (G) being formed as a free acid or as a salt;

[0024] 5) treating compound (G), either as a free base or as a salt, with acid to give amine compound (H), compound (H) being formed as a free acid or as a salt;

[0025] 6) treating compound (H) with l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, hexafluorophosphate azabenzotri azole tetramethyl uronium, or diphenylphosphinic chloride to give the cyclized compound of Formula (I).

[0026] In a second aspect of the invention, the invention provides a process for preparing compound (D), or salt thereof comprising the steps of

[0027] 1) reacting compound (A) and compound (B) in the presence of a base 2) with or without isolating compound (C), contacting Compound (C) with an asymmetric transamination (ATA) enzyme for a time to yield compound (D)

[0028] 3) optionally isolating compound (D) as a salt.

[0029] In a third aspect of the invention, there is a process for preparing a compound of formula (I), comprising the steps of

[0030] 1) contacting compound (D), either as a free base or a salt, with compound (E)

[0031] 2) with or without isolating compound (F), treating compound (F) with a base to give

[0032] 5) treating compound (G) with an acid to give amine compound (H)

[0033] 4) treating compound (H) with l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride to give the cyclized compound of Formula (I).

[0034] In a fourth aspect of the invention, there is provided a process for preparing a compound of formula (I), comprising treating compound (H) with l-ethyl-3-(3-dimethylaminopropyl)carbodiimide either as a free base or as a hydrochloride salt

[0035] In another aspect of the invention, there is provided a process for preparing a compound of formula (I), comprising treating compound (H) with l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide, either as the free base or the hydrochi oridesalt, to give the cyclized compound of Formula

[0036] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the asymmetric transamination (ATA) enzyme used in preparing compound (D) is SEQ ID 1. Alternatively, the compound (D) can be prepared by using the transamination enzyme selected from, but not limited to, Codexis® ATA-025, Codexis® ATA-412, or Codexis® ATA-415. These enzymes are available from Codexis ® and are available from the ATA screening kit. The enzymes may be utilized in a powder form or in a solution form.

[0037] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the peptide coupling reagent used in preparing the compound of Formula (I) is l-ethyl-3-(3- dimethylaminopropyl)carbodiimide either as the free base or as the hydrochloride salt.

[0038] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (D) is formed and isolated as a salt.

[0039] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (D) is formed and isolated as a tartaric acid, dibenzoyl tartaric acid, oxalic acid, malonic acid, succinic acid, fumaric acid, sulfuric acid, hydrobromic acid, or hydrochloric acid salt.

[0040] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (D) is formed and isolated as the D-tartaric acid salt.

[0041] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, compound (D) is the D-tartaric acid salt of compound (D).

[0042] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the reaction of compound A and B to give compound C may be performed in the presence of a base such as CS2CO3, K3PO4, K2CO3, KOH, NaOH, KOt-Bu, DBU, DIPEA, or KHMDS; in another embodiment, the reaction is run in the presence of K3PO4.Gr KOH. The reaction is run in an organic solvent such as acetonitrile, THF, toluene, DMF, or MTBE / water; in another embodiment, the reaction is run in acetonitrile. The reaction may be run at approximately 20- 55 °C, in another embodiment, the reaction is run at room temperature. The reaction is run until compounds (A) and (B) are consumed or (C) is generated. In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the reaction of compound (C) to form compound (D) may be performed using an asymmetric transamination enzyme (ATA). An asymmetric transamination enzyme is an enzyme used to prepare chiral amines in high enantiomeric purity. An asymmetric transamination enzyme can catalyze an asymmetric reductive amination of carbonyl compounds to generate chiral amines. The reaction is typically performed in a buffer to maintain the pH at 6-10, in another embodiment the pH is 8, in another embodiment, the reaction is run in a 0.1 M PB buffer at pH of 8, in another embodiment, the reaction is run in a borate buffer having pH 9.5. The reaction may be run at approximately 20-60 °C, in another embodiment, the reaction is run at approximately 55 °C . There are various ATA enzymes that may be used, including, but not limited to, Codexis® ATA-025, Codexis® ATA-412, or Codexis® ATA-415 or the ATA enzyme SEQ ID. 1.

[0043] Following the formation of compound (D), the compound (D) may be converted to a salt. The compound (D) may be converted to a tartaric acid, dibenzoyl tartaric acid, oxalic acid, malonic acid, succinic acid, fumaric acid, sulfuric acid, hydrobromic acid, or hydrochloric acid salt, alternatively, compound (D) is converted to the D-tartaric acid salt. The salt of compound (D) is then isolated for further reaction. Alternatively, the D- tartartic acid salt of compound (D) is then isolated for further reaction.

[0044] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (D) (as a salt; alternatively as a tartaric acid, dibenzoyl tartaric acid, oxalic acid, malonic acid, succinic acid, fumaric acid, sulfuric acid, hydrobromic acid, or hydrochloric acid salt; or alternatively, the compound (D) as the D-tartaric acid salt) may then be reacted with compound (E) (the synthesis of compound (E) has been previously described) in the presence or absence of an amine, such as DBU, TMG, DMAP, pyridine, NMM, or DIPEA; in another embodiment, the reaction is run in the presence of N,N- diisopropylethylamine. The reaction is run in a solvent, such as IP A, Toluene, EtOH, MeOH, THF, or MeTHF, the reaction may be run with or without water present; in another embodiment, the reaction is run in isopropyl alcohol. The reaction may be run at about 60-82 °C, in another embodiment the reaction is run at approximately 70 °C . Compound (F) may used in the next step without further isolation from the reaction. In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (F) may be reacted in the solution from the above paragraph, or in a separate reaction using solvents such as IP A, toluene, EtOH, MeOH, THF, or MeTHF, with aqueous base, such as NaOH, LiOH, KOH, TMAOH, or TBAOH, in another embodiment the reaction is with NaOH. The reaction is run at a temperature of approximately 60-90 °C, in another embodiment, the reaction is run at a temperature of approximately 75 °C, to hydrolyze the ester group. In one embodiment, toluene is added after the reaction is complete to isolate compound (G) as a toluene solvate.

[0045] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (G) is reacted with an acid to give the unprotected amine, compound (H). In one embodiment, the acid is HC1, TFA, MSA, HBr, H2SO4, or pTSA, in solvents such as methanol, ethanol, IP A, water, MeTHF, TFE, TFA, AcOH, or MeCN, the solvents being used either alone or in combination with each other; in another embodiment, the acid is HC1 in ethanol, dioxane, or water and IP A, in another embodiment the acid is concentrated aqueous HC1 in acetonitrile. The reaction may be run at about 0-50 °C, alternatively, about 20-50 °C, in another embodiment, the reaction is run at about 20 °C until complete. Compound (H) may be isolated as the di-HCl salt, the di-HCl salt of compound (H) may be isolated in either the anhydrous or monohydrate forms.

[0046] In another aspect of the invention, along with the first aspect, second aspect, the third aspect, the fourth aspect, or any other embodiments or aspects, the compound (H) is reacted with a peptide coupling reagent, such as, but not limited to, EDC, EDC / HOBt, HATU, TCFH, T3P, CDI, Vilsmeier reagent, or DPPC1; in another embodiment, the compound (H) is reacted with the peptide coupling reagent l-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride; in another embodiment, the compound (H) is reacted with the peptide coupling reagent l-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride in the presence of 1- hydroxybenzotriazole. The reaction may be run in a solvent such as THF, acetonitrile, DMF, NMP, DMAc, Water, Water+TPGS-750-M surfactant, EtOAc, alternatively, the solvent may be THF, or DMF, or a combination of these solvents. The reaction is run in the presence of an amine, such as DIPEA, NMI, DBU, tert-butylTMG, NMM, pentamethylpiperidine, 2,6-lutidine, 1,4-dimethylpiperazine, or N,N- diisopropylethylamine; in another embodiment, the amine is A,A-diisopropylethylamine. The reaction may be run at approximately -10 to 45 °C, in another embodiment, the reaction is run at approximately 2 °C.

[0047] There are several advantages of preparing the compound of Formula (I) using the present invention.

[0048] Preparing the compound of Formula (I) from compound (H) using the present invention eliminates the use of pentafluorophenyl diphenylphosphinate (FDPP). The reagent FDPP is challenging to source and may not be readily available at all times, but the reagent l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride is readily available. Additionally, the use of FDPP in this reaction produces byproducts which are challenging to remove from the final product. To remove these byproducts, a second crystallization is required. The elimination of the use of FDPP eliminates the need for the second crystallization.

[0049] The process of preparing the compound of Formula (I) from compounds (D) and (E) provides a good overall yield on a large scale, reduced cycle time, less waste, controlled crystallizations, homogeneous reactions, and streamlined work-ups.

[0050] The process of preparing compound (G) from compound (F) affords material properties that are more amenable to different isolation technologies and obviates the risk of filter cloth binding. Compound (G) is formed as a neutral solvate which provides advantages in isolation.

[0051] The process of preparing compound (H) from compound (G) has the advantage that it produces a single form in a controlled way and via an alternate reaction / crystallization conditions. The isolation of the single consistent form is not influenced by isolation or drying equipment and parameters, and it is amenable to different isolation technologies, e.g., centrifugation.

[0052] The process of preparing compound (D) produces the D-tartrate salt which is readily isolated. The process of preparing compound (D) is 2 steps from compound (A), providing good overall yield, lower cycle time, and lower costs.

[0053] The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. This invention encompasses all combinations of preferred aspects and / or embodiments of the invention noted herein. It is understood that any and all embodiments of the present invention may be taken in conjunction with any other embodiment or embodiments to describe additional embodiments. It is also to be understood that each individual element of the embodiments is its own independent embodiment. Furthermore, any element of an embodiment is meant to be combined with any and all other elements from any embodiment to describe an additional embodiment.

[0054] DEFINITIONS

[0055] To facilitate understanding of the disclosure set forth herein, a number of additional terms are defined below. Generally, the nomenclature used herein and the laboratory procedures in organic chemistry, medicinal chemistry, and pharmacology described herein are those well-known and commonly employed in the art. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0056] Unless specifically stated otherwise herein, references made in the singular may also include the plural. For example, “a” and “an” may refer to either one, or one or more.

[0057] The term “about” or “approximately” is intended to indicate that the exact time or temperature is not required. For example, and not to limit the invention, when used with a temperature, it is intended to cover 5 °C above or below the indicated range.

[0058] EXAMPLES

[0059] The following Examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below were performed or that they are all of the experiments that may be performed. It is to be understood that exemplary descriptions written in the present tense were not necessarily performed, but rather that the descriptions can be performed to generate data and the like of a nature described therein. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for. The skilled artisan will recognize a variety of analytical methods that can be used to characterize the compounds described herein, including, for example,JH nuclear magnetic resonance spectroscopy (NMR), heteronuclear NMR, mass spectrometry (MS), liquid chromatography (LC), and infrared (IR) spectroscopy. The foregoing list is a subset of characterization methods available to a skilled artisan and is not intended to be limiting.

[0060] To further illustrate the foregoing, the following non-limiting, exemplary synthetic schemes are included. Variations of these examples within the scope of the claims are within the purview of one skilled in the art and are considered to fall within the scope of the invention as described and claimed herein. The reader will recognize that the skilled artisan, provided with the present disclosure, and skill in the art is able to prepare and use the invention without exhaustive examples.

[0061] The following abbreviations have the indicated meanings:

[0062] / / mol - micromole

[0063] AcOH = acetic acid aq = aqueous

[0064] ATA - amine transaminase

[0065] MeCN or CH3CN = acetonitrile

[0066] CDI = carbonyldiimidazole dd = doublet of doublet ddd = doublet of doublet of doublets

[0067] DBU = l,8-Diazabicyclo[5.4.0]undec-7-ene

[0068] DIPEA = 7V,7V-Diisopropylethylamine

[0069] DMAc = dimethylacetamide DMAP = dimethylaminopyridine DMF = dimethylformamide DMSO = dimethylsulfoxide DPPC1 = diphenylphosphinic chloride EDAc or EDC = l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride equiv = equivalent(s) EtOAc = ethyl acetate EtOH = ethanol g = gram(s) h = hour

[0070] HATU = Hexafluorophosphate Azabenzotri azole Tetramethyl Uronium

[0071] HC1 = hydrogen chloride (usually as a solution)

[0072] H2O = water

[0073] HOBt = Hydroxybenzotriazole

[0074] HPLC = High Pressure Liquid Chromatography

[0075] IPA = isopropyl alcohol kg = kilogram

[0076] KOH = potassium hydroxide

[0077] L = liter

[0078] LiOH = lithium hydroxide m = multiplet

[0079] M = molar

[0080] MeOH = methanol mg = milligram(s)

[0081] MeOH or CH3OH = methanol

[0082] MeTHF = methyl tetrahydrofuran

[0083] MHz = megahertz min = minute(s) mL = milliliter

[0084] MTBE = methyl tert-butyl ether mmol = millimole(s)

[0085] MSA = methanesulfonic acid

[0086] NaOH = sodium hydroxide

[0087] NH40H - ammonium hydroxide

[0088] NMI = 1 -Methylimidazole

[0089] NMM = A-Methylmorpholine

[0090] NMP = n-methylpyrrolidine

[0091] PLP = pyridoxal phosphate pTSA = p-toluenesulfonic acid q = quartet s = singlet t = triplet

[0092] T3P = Propanephosphonic acid anhydride

[0093] TBAOH = Tetrabutylammonium hydroxide

[0094] TCFH = chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate td = triplet of doublets tert-butylTMG = 2-tert-Butyl-l,l,3,3-tetramethylguanidine

[0095] TFA = trifluoroacetic acid

[0096] TFE = trifluoroethanol

[0097] THF = tetrahydrofuran

[0098] TPGS = tocopheryl polyethylene glycol succinate

[0099] TMAOH = tetramethylammonium hydroxide

[0100] °C = degrees Celsius

[0101] UPLC / UHPLC - ultra performance liquid chromatography

[0102] Vilsmeier reagent = (Chlormethylene)dimethylammonium chloride vol = volumes wt = weight

[0103] The invention is further defined in the following Example. It should be understood that the Example is given by way of illustration only. From the above discussion and the Example, one skilled in the art can ascertain the essential characteristics of the invention, and without departing from the spirit and scope thereof, can make various changes and modifications to adapt the invention to various uses and conditions. As a result, the invention is not limited by the illustrative examples set forth herein below, but rather is defined by the claims appended hereto. Step 1

[0104] D (D-tartrate)

[0105] Step 1

[0106] To a solution of compound (A) (12.0 g, 50.6 mmol, 1.0 equiv) and compound (B) (8.18 g, 53.2 mmol, 1.05 equiv) in acetonitrile (60 mL) was added solid K3PO4 (16.2 g, 76.8 mmol, 1.5 equiv), followed by tetrapropylammonium chloride (1.14 g, 5.06 mmol, 0.10 equiv) at 20 °C under atmosphere of nitrogen. The mixture was stirred vigorously until sulfonamide was consumed as determined by HPLC analysis. Subsequently MTBE (60 mL) was added, and the resulting mixture was cooled to 0-5 °C. Aq. HC1 solution (1.0 N, 76 mL) was slowly added at <20 °C. The mixture was then warmed to 20 °C, and water (120 mL) was added to dissolve the solids. The biphasic mixture was aged until the sulfonic acid intermediate was consumed as determined by HPLC analysis. The aq. phase was then removed. Heptane (96 mL) was added into the organic phase, and the resulting solution was then washed with 4% aq. NaOH solution (72 mL x2) and 5% aq. KH2PO4 solution (24 mL), subsequently solvent exchanged to EtOH. The batch volume was adjusted to ca. 30 mL, and was then cooled to 0-5 °C. Water (6.0 mL) was added over a period of 15 min, and the mixture was stirred until the seed bed was formed. Additional water (36 mL) was added over a period of 1 h, and the resulting slurry was stirred for 3 h at 0-5 °C prior to filtration. The filtrate was recycled to complete the transfer of the slurry in the reactor. The filter cake was washed with water (24 mL x2) and dried under vacuum at <35 °C, affording 14.0 g of compound (C) in 89.0% yield. 'H NMR (500 MHz, CHLOROFORM-d) 5 7.39 (dd, J=8.9, 3.2 Hz, 1H), 7.13 (ddd, J=9.0, 7.4, 3.3 Hz, 1H), 6.97 (dd, J=9.0, 4.0 Hz, 1H), 5.03 (br s, 1H), 4.63 - 4.52 (m, 1H), 3.50 - 3.40 (m, 1H), 3.38 - 3.29 (m, 1H), 2.60 (s, 3H), 1.43 (s, 9H), 1.33 (d, J=6.1 Hz, 3H). Alternatively, compound (C) may be prepared using the following procedure.

[0107] In a 250mL chemglass reactor under N2, degassed acetonitrile, (80 mL, 4 L / kg) [a range of 3.6-4.4 L / kg MeCN can be used] was combined with l-(5-fluoro-2-hydroxy- phenyl)ethanone (15.59 g, 101.2 mmol, 1.20 equiv) [a range of 1.1-1.3 equiv can be used] and degassed with 3 cycles of vacuum / nitrogen. A solution of KOH (30 mass%) in water (18.92 g, 101 mmol, 1.20 equiv) [a range of 1.1-1.3 equiv can be used] was stirred over the course of ~5 min and allowed to age for 30min. Compound (B) (20.00 g, 84.29 mmol, 1.00 equiv) was added and stirred in a single portion and heated to 30 °C [a range of 25- 40 °C can be used]. After 24 h at 30 °C, sulfuric acid (0.45 mL, 8.4 mmol, 0.10 equiv) [a range of 0.09-0.11 equiv can be used] was added. Water for workup (100 mL5 L / kg) [a range of 4.5-5.5 L / kg can be used] was stirred in over the course of ~5 min. The reaction was stirred vigorously for 30 min at 25 °C, then tert-butyl methyl ether (120 mL, 6 L / kg) [a range of 5.4-6.6 L / kg can be used] was added. The temperature was adjusted to 20 °C; the reaction was stirred 10 minand allowed to settle for 10 min. The aq layer was separated, and the organic layer was washed with 5 wt% aq. KOH (60 mL, 3L / kg) [a range of 2.7-3.3 L / kg can be used] twice. The aqueous layer was separated. The organic layer was washed with 5 wt% aq. potassium dihydrogen phosphate (60 mL, 3 L / kg) [a range of 2.7-3.3 L / kg can be used], and then the aq layer was separated. The organic layer was stirred with activated carbon (Darco 100 mesh, 3.0 g, 3.0 equiv) for 15 min at 22 °C. The mixture was filtered, and the filter cake was rinsed with 20 mL MTBE. The filtrate was concentrated to a final volume of 60 mL (3 L / kg) at 280 mbar with jacket temp up to 50 °C. 100 mL of EtOH (5 L / kg) [a range of 4.5-5.5 L / kg can be used] was stirred in and redistilled back to ~60 mL (3 L / kg) [a range of 2.7-3.3 L / kg can be used] at 180 mbar with jacket temp up to 60 °C. EtOH was added to dilute the reaction to a total volume of 120 mL (6 L / kg), which was then cooled to -5 °C (jacket set to -20 °C) [a range of -10-5 °C can be used]. Water (60 mL, 3 L / kg) [a range of 2.7-3.3 L / kg can be used] was stirred in over the course of 15 min prior to seeding. The jacket temp was adjusted to -9 °C, and the reaction was seeded with seeds of compound D (200 mg) [a range of 0.9-1. lwt% can be used]. The mixture was aged for 20 min. Water (90 mL, 4.5 L / kg) [a range of 4.0-5.0 L / kg can be used] was stirred in dropwise over the course of ~30 min. The mixture was aged for 15 h at -2 °C. The batch was filtered and the reactor and cake were rinsed with cold (0 °C) 2: 1 water: EtOH (60 mL, 3 L / kg) [a range of 2.7-3.3 L / kg can be used], then the cake was rinsed with water (60 mL, 3 L / kg) [a range of 2.7-3.3 L / kg can be used]. The solid was dried under vacuum with N2 purge for 48 h at 25 °C, affording 22.93 g (86.5% yield as-is) as a white powder.

[0108] Step 2

[0109] 0.1 M phosphate buffer pH 8.0 (70 mL, 14 vol), 6 M isopropylamine hydrochloride (65 mL, 25 eq, 13 vol), PLP (0.5 g, 10 wt%) and ATA enzyme having the enzyme SEQ ID NO 1 (as shown below) (0.5 g, 10 wt%) were charged into the reactor and dissolved by stirring. The compound (C) (5.0 g, LR) was dissolved in DMSO (40 mL, 8 vol), and the solution was added to the enzyme solution. The reaction mixture was stirred at 55°C for 24 hours. After reaction completion, celite 454 (10 g) was added to the reaction mixture, and the solution pH was adjusted to 9-10 by addition of aq KOH solution (5%). MTBE (100 mL, 20 vol) was added into the batch, and the resulting mixture was agitated for 0.5 h. The batch was filtered, and the reactor and filter cake were rinsed with MTBE (15 mL). The aq phase was removed from the combined filtrates, and was extracted with MTBE (50 mL, 10 vol). The combined organic phases were washed with 15% aq NaCl solution (15 mL, 3 vol) and solvent-swapped to isopropyl alcohol (IP A). The batch volume was adjusted to ~17 mL (~3.5 vol). A solution of D-tartaric acid (2.52 g, 1.05 eq) in IPA (58 mL) was added into the batch. The batch was warmed to ~80 °C until a clear solution was observed. MTBE (5 mL, 1 vol) was added, and the batch was cooled to 0-5 °C over a period of 15 h. The batch was stirred at 0-5 °C for >2 h and filtered. The reactor was rinsed with MTBE (12.5 mL, 2.5 vol), and the rinse was applied to the cake wash. The cake was dried at 50 °C under vacuum to yield compound (D) as a white solid in 87% yield (6.7g, 99.4wt%). 'H NMR (500 MHz, DMSO-d6) 8 7.31 (dd, J=9.4, 2.7 Hz, 1H), 7.15 - 7.11 (m, 2H), 4.63 (q, J=6.6 Hz, 1H), 4.49 (sxt, J=5.8 Hz, 1H), 3.92 (s, 2H), 3.29 - 3.12 (m, 2H), 1.43 (d, J=6.7 Hz, 3H), 1.38 (s, 9H), 1.19 (d, J=6.Q Hz, 3H).

[0110] SEQ ID Nol : MTTTEFANREFH(12aa)MTTTEFANSNLVAVEPVAIREPTPPGSVIQYSEYELDRSH PLAGGVAWIEGEYVPADEARISIFDMGFYCSDATYTGIHVWHGNIFRLEDHLDRL LHGAARLKLETGMSREELAGIAKRCVSLSQLREAMVNITITRGYGSTPYGRDATK HRPQVYVYAGPYQWIFPPEEQIFGTSVIVPRHVRRAGLNTIDPTIKNFQWGDLSAA IREAHDRGARSAVLLDADNCVAEGPGFNVVLVKDGALVSPSRNALPGITRKTVY EIAAAKGIETMLRDVTSKELYEADELMAVSAAGGVTPITSLDGEQVGNGEPGPIT VAIRDRFWALMDEPSSLIEAIDY.

[0111] There are 12 amino acids ahead of the transaminase sequence in SEQ ID NO: 1.

[0112] Alternatively, Step 2 can be performed as follows.

[0113] A 0.1 M borate buffer pH 9.5 (125 mL, 25 vol), isopropylamine hydrochloride (15.4g, 10 equiv), PLP (0.5g, 10 wt%) and ATA enzyme having SEQ ID NO1 (0.5 g, 10 wt%) were charged into the reactor and dissolved by stirring. Compound (C) (5.0 g, LR) was dissolved in DMSO (25 mL, 5 vol), and the solution was added to the enzyme solution. Note: precipitation formation was observed due to poor solubility of compound C in aq buffer. The reaction mixture was stirred at 45 °C for 48 hours with continuous N2 bubbling. After the reaction was complete, NaCl (50 g) and t-amyl alcohol (75mL, 15 vol) were added, and the reaction mixture was stirred overnight. Celite (5 g, 100wt%) was added , and the resulting mixture was agitated for 2 h. The batch was filtered, and the filter cake was reslurried with 1 : 1 water: t-amyl alcohol (25 mL, 5 vol) twice. The filtrates were combined and adjusted the pH to 10-11 by aq NaOH. This was stirred for Ih. The organic layer was separated, and then the aq layer was back extracted with t-amyl alcohol (25 mL, 5 vol). The organic layers were combined and concentrated to a final volume of 6.5 mL (1.3 vol) at 50 °C and then 7:3 MTBE:Heptane (50 mL, 10 vol) was added. The stream was washed with 15wt% aq NaCl solution (25 mL, 5 vol) three times. The solvent was swapped to t-amyl alcohol and then concentrated to a final volume of 17.5mL (3.5vol) at 50 °C. A solution of D-tartaric acid (2.52 g, 1.05 eq) in t-amyl alcohol (82.5 mL, 16.5vol) was added into the batch. The batch was warmed to ~80 °C and stirred at 80 °C for 4 hours. MTBE (15 mL, 3 vol) was added at 80 °C, and the batch was cooled to 0-5 °C over a period of 15 h. The batch was stirred at 0-5 °C for >2 h and filtered. The reactor was rinsed with MTBE (12.5 mL, 2.5v), and the rinse was applied as a cake wash. The cake was dried at 50 °C under vacuum to provide Compound D tartrate product as a white solid in 75% yield (5.78 g, 99.4wt%). Alternatively, Step 2 can be performed as follows:

[0114] Triethanolamine hydrochloride (0.6 g) and pyridoxyl-L-phosphate (8.0 mg) were dissolved in 20 mL of water. To this solution was added isopropylamine (2.5 mL) or isopropylamine hydrochloride (2.9 g), and the pH was adjusted to pH 8.0 using 5 M NaOH or HC1 (Solution A). Compound C (150 mg) was dissolved in DMSO (3 mL). Solution A (450ul) was added to each vial containing 1 mg of the ATA enzyme and agitated on a shaker for 10-15 minutes to ensure all the enzyme is dissolved. Solution of Compound (C) (50ul) was added to each vial of ATA enzyme. The vials were sealed and agitated at 600 RPM at 30 °C for 18 hours.

[0115] Compound (C) was prepared using this process and the ATA enzymes Codexis® ATA-025, Codexis® ATA-412, or Codexis® ATA-415. Yields of compound C was 2-

[0116] 15% using HPLC analysis.

[0117] To a reactor containing compound (D) (D-tartrate) (LR; 250 g) was added isopropanol (IP A; 4.0 L / kg; 1000 mL; 786 g) [A range of 3.6-4.4 L / kg IPA can be charged]. To the resulting slurry was added 5 wt% aqueous NaOH (1.6 equiv.; 657 mL; 692 g) [A range of 2.37-2.89 L / kg 5 wt% NaOH can be charged]. The resultant solution was stirred for 30 minutes at a temperature range of 10-35 °C, over which time it became a slurry. N,N-Diisopropylethylamine (DIPEA; 2.5 equiv.; 236 mL; 175 g) was added [a range of 2.25-2.75 equiv DIPEA can be charged], followed by IPA (0.25 L / kg; 62.5 mL; 49.1 g) as a rinse. Compound (E) (1.10 equiv.; 134.2 g) [a range of 1.03-1.21 equiv E can be charged] was then added, and IPA (0.75 L / kg; 188 mL; 147 g) was again added as a rinse. The subsequent slurry was warmed to 70 °C for 12 hours (a range of 60-79 °C, with longer reaction times needed for lower temperature). The reaction was cooled to 40 °C and sampled for conversion to compound (F) as judged by UPLC. Compound (F) was then directly transformed without isolation. To the reaction was added 25 wt% aqueous NaOH (10.0 equiv.; 679 mL; 865 g) [a range of 2.43-2.97 L / kg of 25 wt% aqeous NaOHcan be charged] and the biphasic solution was then warmed to 70 °C for 12 hours (a range of 65-79 °C, with longer reaction times needed for lower temperature). After cooling to 40 °C and sampling for conversion to Compound (G) as judged by UPLC, 2- methyltetrahydrofuran (2-MeTHF; 5.0 L / kg; 1250 mL; 1070 g) [a range of 4.5-5.5 L / kg 2-MeTHF can be charged) and water (4.0 L / kg; 1000 mL; 1000 g) [a range of 3.6-4.4 L / kg water can be charged] were added, and the aqueous layer discarded. The organic phase was then iteratively washed with 20 wt% aqueous NaCl (7.0 L / kg; 1750 mL; 2010 g) [a range of 6.3-7.7 L / kg can be charged] followed by 20 wt% aqueous citric acid (3.0 L / kg; 750 mL; 814 g) [a range of 2.7-3.3 L / kg 20 wt% aqueous citric acid can be charged]. Toluene (10.0 L / kg; 2500 mL; 2180 g) [a range of 9-11 L / kg toluene can be charged] and water (1.0 L / kg; 250 mL; 250 g) [a range of 0.9-1.1 L / kg water can be charged] were then added, and the biphasic mixture was stirred for 30 minutes. After discarding the aqueous layer, the solution was distilled under reduced pressure (100-300 mBar) to a residual volume of 5.95-8.05 L / kg (end point: 1750 mL). At 50 °C (range of 45-60 °C), the solution was then seeded with 1.0 wt% (a range of 0.25 - 2.15 wt% seeds can be charged) compound (G). Following the addition of toluene (8 L / kg; 2000 mL;

[0118] 1740 mL) [a range of 7.2-8.8 L / kg can be charged], cooling, and aging overnight, the slurry was filtered and washed twice with toluene (3 L / kg; 750 mL; 653 g) [a total of 2-12 L / kg toluene can be charged] to afford (G) (as a toluene solvate) (248 g, 91% potency, 9% toluene; 87% adjusted yield) as a white powder.JH NMR (DMSO-de) 8 11.50 (br s, 1H), 8.56 (d, 1H), 8.31 (d, 1H), 8.11 (s, 1H), 7.24 (m, 1H), 7.16 (m, 1H), 7.16 (toluene, m, 1H) 7.02 (toluene; m, 4H), 6.80 (m, 1H), 6.46 (d, 1H), 5.69 (m, 1H), 4.46 (m, 1H), 3.5-3.2 (m, 2H), 3.1 (m, 1H), 2.30 (toluene, s, 3H), 1.53-1.14 (m, 15H)

[0119] (toluene solvate)

[0120] Compound (G) (10.0 g, 1 equiv, 21.1 mmol), acetonitrile (50.0 mL) [a range of 4-6 L / kg acetonitrile can be charged], and water (10.0 mL) [a range of 0.70-1.00 L / kg can be charged] were added in sequence to a 100 mL Chemglass reactor at 20 °C. The resulting mixture was cooled to 15 °C (range of 5-35 °C), and then aqueous hydrochloric acid (36- 37 wt% solution, 13.0 mL, 7.40 equiv, 156 mmol) [a range of 6-8 equiv 33 wt% HCI can be charged] was slowly added over 30 min. When the addition of HCI aq. was complete, compound (H) (50 mg, 0.005 equiv, 108 / / mol) [a range of 0.75-1.25 wt% seeds can be charged] was added, and the resulting mixture was warmed to 20 °C (range of 12-35 °C, with longer reaction times needed for lower temperature) and aged for 18 h at 20 °C. When the reaction was complete by HPLC analysis, acetonitrile (70 mL) [a range of 6-8 L / kg acetonitrile can be charged] was added in one portion to the product mixture. The resulting mixture was allowed to age for no less than 2 h at 20 °C. When the aging process was complete, the solids were collected by vacuum filtration and then washed with acetonitrile (50.0 mL) [a total range of 3-9 L / kg acetonitrile can be charged]. The collected solids were then dried under reduced pressure and at 50 °C for 18 h to afford compound (H) as a white solid (8.3 g, 17.9, 85%).1H NMR (400 MHz, trifluoroacetic acid- / / ) 5 9.27 - 8.89 (m, 1H), 8.77 - 8.32 (m, 1H), 7.86 - 6.77 (m, 7H), 5.99 - 5.54 (m, 1H), 5.30 - 5.05 (m, 1H), 3.78 (s, 2H), 2.10 - 1.79 (m, 3H), 1.66 - 1.34 (m, 3H). *some of the proton signals from N-H / OH based functional groups could not be visualized. p l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (5.16 g, 26.9 mmol, 1.25 equiv) [a range of 1.1-1.4 equiv l-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride can be charged] and 1- hydroxybenzotriazole (20% wet) (1.09 g, 6.5 mmol, 0.30 equiv) [a range of 0.24-0.36 equiv HOBt can be charged with a range of 12-20 wt% water content] were added to a solution of tetrahydrofuran (25 mL, 2.5 L / kg) [a range of 2-3 L / kg THF can be charged] plus N,N-dimethylformamide (25 mL, 2.5 L / kg) [a range of 2-3 L / kg DMF can be charged]. The resultant slurry was cooled to -5 °C, and N,N-diisopropylethylamine (8.5 mL, 48.4 mmol, 2.25 equiv) [a range of 2.02-2.48 equiv DIPEA can be charged] was added, followed by compound (H) (10.0 g, 21.5 mmol, limiting reagent). The resultant mixture was aged for 24 h at an internal temperature of 3 °C (range of -10 to 30 °C, with longer reaction times needed for lower temperature). (Optionally, 25 wt% aq NH4OH (0- 0.6 equiv) can be charged as a quench followed by 1 h age (not charged in this procedure.)) Next, water (60 mL, 6 L / kg) [a range 5.4-6.6 L / kg water can be charged] was added, and the resultant solution was heated to 35 °C (range of 27-42 °C). Seeds of the compound of Formula (I) (100 mg, 0.01 g / g) [a range of 0.25-1.75 wt% seeds can be charged] were added followed by a 2 h age. Additional water (80 mL, 8 L / kg) [a range of 7-9 L / kg water can be charged] was charged slowly over the course of 2 h, followed by 2 h of additional age at 35 °C. The slurry was cooled to 20 °C over the course of 2 h, aged for 16 h more, then filtered. The filter cake was washed with 3: 1 water: tetrahydrofuran (40 mL, 4 L / kg) [a range of 3.2-4.8 L / kg 3: 1 watertetrahydrofuran can be charged], then water (40 mL, 4 L / kg) [a range of 3.2-4.8 L / kg water can be charged], then water (40 mL, 4 L / kg) [a range of 3.2-4.8 L / kg water can be charged]. Vacuum oven drying (24 h at 65 °C and 200 mbar with a slow nitrogen purge) afforded the compound of Formula (I) (6.85 g, 89.5% yield) as a white crystalline solid.JH NMR (400 MHz, DMSO-d6): 9.84 (dd, 1 H, J= 1.6, 8.0 Hz), 8.82 (d, 1 H, J= 6.8 Hz), 8.58 (d, 1 H, J= 7.6 Hz), 8.06 (s, 1 H), 7.15 (dd, 1 H, J= 3.2, 9.6 Hz), 7.05 - 6.94 (m, 2 H), 6.37 (d, 1 H, J= 7.6 Hz), 5.56 (m, 1 H), 4.50 (m, 1 H), 3.93 (ddd, 1 H, J= 3.6, 8.0, 13.2 Hz), 3.16 (ddd, 1 H, J= 2.0, 8.4, 13.6 Hz), 1.48 (d, 3 H, .7= 6.0 Hz), 1.47 (d, 3 H, J = 7.2 Hz).

Claims

WHAT IS CLAIMED IS:

1. A process for preparing a compound of formula (I):comprising the steps of:1) reacting compound A and compound B in the presence of a baseto give compound (C)2) with or without isolating compound (C), contacting compound (C) with an asymmetric transamination enzyme to give compound (D)3) contacting compound (D), either as a free base or as a salt, with compound (E)4) with or without isolating compound (F), treating compound (F) with a base to give5) treating compound G with acid to give an unprotected amine compound (H)6) treating compound (H) with -ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, hexafluorophosphate azabenzotri azole tetramethyl uronium, or diphenylphosphinic chloride to give the cyclized compound of Formula (I).

2. The process of claims 1, whereinCompound (H) is treated with l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride to form the compound of Fomula (I).

3. The process of claims 1-2, wherein in the reaction of compound (D) and (E), compound (D) is present as the salt and a couterion of the salt4. The process of claims 3, wherein in the reaction of compound (D) and (E), compound (D) is present as the tartaric acid salt.

5. A process of preparing compound D, or salt thereofcomprising the steps of1) reacting a compound A and a compound B in the presence of a base2) with or without isolating compound (C), contacting Compound (C) with an asymetric transamination enzyme for a time to yield compound (D)3) optionally isolating compound (D) as the salt.

6. The method of claim 5, wherein the compound (D) is formed and isolated as the D-tartric acid salt.

7. A process of preparing a compound of formula (I),comprising the steps of 1) contacting compound (D), either as the free base or a salt thereof, with compound (E)2) with or without isolating compound (F), treating compound (F) with a base to give5) treating compound G with an acid to give an unprotected amine compound (H)4) treating compound (H) with l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride to give the cyclized compound of Formula (I).

8. The process of claim 7, wherein compound (D) is the D-tartaric acid salt of compound (D).

9. A process for preparinga compound of formula (I),comprising treating compound (H) with l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride to give the cyclized compound of Formula (I)

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