A new synthetic method for the preparation of (6R,10S)-10-{4-[5-chloro-2-(4-chloro-1H-1,2,3-triazol-1-yl)phenyl]-6-oxo-1(6H)-pyrimidinyl}-1-(difluoromethyl)-6-methyl-1,4,7,8,9,10-hexahydro-11,15-(methano)pyrazolo[4,3-B][1,7]diazacyclotetradecin-5(6H)-one

Abstract

A highly efficient method is provided for the preparation of a key intermediate in the synthesis of compound (I), which is broadly applicable and can provide a selection of components with a variety of substituents. TIFF2022526659000174.tif61153

Description

[Technical field]

[0001] The present invention relates generally to several improved methods for the preparation of (6R,10S)-10-{4-[5-chloro-2-(4-chloro-1H-1,2,3-triazol-1-yl)phenyl]-6-oxo-1(6H)-pyrimidinyl}-1-(difluoromethyl)-6-methyl-1,4,7,8,9,10-hexahydro-11,15-(methano)pyrazolo[4,3-b][1,7]diazacyclotetradecin-5(6H)-one, a factor XIa inhibitor useful in the treatment of thromboembolic disorders (e.g., venous thrombosis and deep vein thrombosis). [Background technology]

[0002] Factor XIa is a plasma serine protease involved in the regulation of blood coagulation, which in vivo initiates the binding of tissue factor (TF) to factor VII (FVII) to produce factor VIIa (FVIIa). The resulting TF:FVIIa complex contributes to the activation of factor IX (FIX) and factor X (FX) leading to the generation of factor Xa (FXa). The produced FXa functions as a catalyst to convert prothrombin to small amounts of thrombin, after which this pathway is inhibited by tissue factor pathway inhibitor (TFPI). Catalytic amounts of thrombin provide feedback activity to factors V, VIII, and XI, further propagating the coagulation process (Gailani, D. et al., Arterioscler. Thromb. Vasc. Biol., 27:2507-2513 (2007)). Increased thrombin polymerizes fibrinogen to fibrin, forming the framework of the clot and activating platelets, a key cellular coagulation component (Hoffman, M., Blood Reviews, 17:S1-S5 (2003)). Factor XIa therefore plays a key role in propagating this amplification loop, making it an attractive target for antithrombotic therapy.

[0003] US Patent No. 9,453,018 discloses macrocyclic compounds as factor XIa inhibitors useful for the treatment of thromboembolic disorders. One of the compounds has the following structure: [ka]

[0004] The above-mentioned US patent discloses a multi-step synthetic method for preparing macrocycles, which includes coupling a pyridine-containing macrocycle with a pyrimidinol to form compound (I). The disclosed method also includes a ring-closing metathesis step using a catalyst, such as Grubbs (II).

[0005] There are various difficulties in applying the multi-step synthesis disclosed in US Pat. No. 9,453,018 to larger scale synthesis (e.g., pilot plant or manufacturing scale production). One of them is that the Grubbs (II) reagent is expensive, making it difficult to apply to industrial scale synthesis. Furthermore, there is a continuing need to search for methods that provide higher yields in order to reduce production costs and / or reduce unreacted materials. Preferably, inexpensive starting materials are selected for new processes.

[0006] It would be desirable to have processes suitable for producing larger quantities of compound (I) than are typically produced in laboratory scale processes, and processes which provide compound (I) in higher yields than previously disclosed processes.

[0007] The present invention is directed to either or both of these aspects, as well as other important aspects. Summary of the Invention

[0008] In one embodiment, the present invention provides a method for preparing compound (I), characterized by the steps of: [ka] 1) Formula [ka] Compound 1 having the structure of the formula [ka] [In the formula, X is selected from Cl, Br, and I; Y is OR 9 , N.H.O.C. 1-3 alkyl, Cl, Br, and I; and R 9 is C 1-3 Alkyl, C 1-3 hydroxyalkyl, substituted phenyl and substituted benzyl; in a suitable solvent to give a compound of formula [ka] Compound 3a or 3b is obtained. 2) reacting compound 3a or 3b with a compound of formula [ka] is converted to compound 4. 3) Next, compound 4 is reacted with trialkyl orthoformate in an alcohol solvent to obtain the compound of the formula [ka] [In the formula, R 1 is C 1-6 is alkyl; and R 3 ' is C 1-6 alkyl, optionally substituted phenyl and benzyl] to obtain compound 5. 4) The ester of compound 5 is hydrolyzed under basic conditions or R 3 When ' is a substituted benzyl, it undergoes hydrogenolysis, [ka] In the formula, R 1and compound 6, where X is defined as above. 5) The carboxyl moiety of compound 6 is then activated and reacted with a chiral auxiliary to give the compound of formula [ka] [In the formula, Xa is a chiral auxiliary, and R 1 and X is defined as above] to obtain compound 6a. 6) Compound 6a is then reacted with a base in the presence of a methyl donor (e.g., an alkyl halide) and a chiral auxiliary; [ka] In the formula, R 1 Compound 6b is obtained, in which X and Xa are defined as above. 7) Removal of the chiral auxiliary Xa; [ka] In the formula, R 1 and to obtain compound 7, in which X is defined as above. 8) Next, compound 7 is reacted with the compound having the following structure in the presence of a metal catalyst: [ka] React with compound 8 to give compound 9 [ka] get. 9) The nitro group of compound 9 is reduced to give the compound of the following formula: [ka] Compound 10 is obtained. 10) Compound 10 is cyclized with a suitable coupling agent to give compound 11 [ka] get. 11) The ketone group is deprotected in the presence of acid to give compound 12. [ka] get. 12a) reducing compound 12 with an equivalent amount of ammonia in the presence of a reducing agent; or 12b) Compound 12 is reduced by aminotransferase in the presence of an amine donor, a different recyclization system, and a cofactor to give compound 13. [ka] This gives rise to an amine stereocenter represented by the formula: 13) Compound 13 is then reacted with compound 14 [ka] to give compound (I). [ka] get.

[0009] In yet another aspect, the present invention relates to a compound of formula (II): [ka] [In the formula, The dashed lines (----) are optional bonds; R 1 is C 1-6 alkyl, preferably Me; R 2 is C 1-3 alkyl, preferably Me; R 3 OH, O.C. 1-4 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I. or a free base form or salt thereof.

[0010] In some embodiments of the compound of Formula (II), R 1 is C 1-6 is alkyl; R 2 is C 1-3 is alkyl; R 3 OH, O.C. 1-6 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3 selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I.

[0011] In some embodiments of the compound of Formula (II) or a pharma- ceutical acceptable salt thereof, R 1 is methyl; R 2 is methyl; R 3 is OH; and X is Cl.

[0012] In some embodiments of the compound of Formula (II), R 1 is methyl; R 2 is methyl; R 3 is OH; and X is Cl; The compound is in the form of its free base or as the (1S,2R)-2-amino-1,2-diphenylethan-1-ol salt or dicyclohexylamine salt.

[0013] In yet another aspect, the present invention relates to a compound of formula (IIa): [ka] [In the formula, The dashed lines (----) are optional bonds; R 1 is C 1-6 alkyl, preferably Me; R 2 is C 1-3 Alkyl or C 1-3 alkenyl, preferably Me or CH2; R 3 OH, O.C. 1-4 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3 selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I. or a free base form or salt thereof.

[0014] In some embodiments, the compound of formula (II) has the structure: [ka] [In the formula, R 1 is methyl; R 2 is methyl; R 3 is OH; and X is Cl. The method for producing the compound of is characterized by the following steps: a) Formula [ka] Compound 22 of the formula [ka] Compound 2 [In the formula, X is selected from Cl, Br, and I; Y is OR 9 , N.H.O.C. 1-3 alkyl, Cl, Br, and I; and R 9 is C 1-3 Alkyl, C 1-3 hydroxyalkyl, substituted phenyl and substituted benzyl. in a suitable solvent to give a compound of the formula [ka] Compound 23 is obtained. b) reacting compound 23a or 23b with a compound of formula [ka] to compound 24. c) Compound 24 is then reacted with trimethyl orthoformate or triethyl orthoformate to give the compound of formula [ka] [In the formula, R 1 is methyl or ethyl, and X is as defined above to obtain compound 25. d) Compound 25 is enzymatically [ka] In the formula, R 1and converted to compound 26, wherein X is defined as above. e) hydrolyzing compound 26; [ka] In the formula, R 1 and converted to compound 27, wherein X is defined as above.

[0015] In some embodiments of the process for making a compound of Formula (II), the enzyme is a lipase.

[0016] In some embodiments, the structure of compound 21 [ka] The process for the preparation of compounds of formula (II) or (IIa) having the formula (II) is characterized by the following steps: 1) Formula [ka] R is C 1-6 The alkyl cyclopentane ester derivative is reacted with a dialkylamine to give a compound of the formula [ka] Alk is C 1-6 To obtain compound 40, which is an alkyl group. 2) Compound 40 is combined with a first base to form the following structure: [ka] to form compound 41. 3) Compound 41 is reacted with an acid and an alcohol R 3 'OH to give the formula [ka] R 3 ' is C 1-6 alkyl, forming compound 42. 4) Compound 42 is synthesized by the following method. [ka] in the presence of a second base to obtain a compound of the formula [ka] Compound 18b is obtained. 5) Compound 18b is synthesized by the method according to the following structure: [ka] is converted to compound 20b. 6) Compound 20b is then hydrogenated to give the following structure: [ka] to obtain a compound of formula (II) having the formula

[0017] In some embodiments of the process for preparing a compound of Formula (II) or (IIa) having the structure of compound 21, the hydrogenation in step (6) comprises a chiral ruthenium catalyst.

[0018] formula [ka] [In the formula, R1 is C 1-6 is alkyl; R 10 is C 1-6 is alkyl] In some embodiments, the method for preparing a compound of formula (II) or (IIa) having compound 19 of the formula (II) is characterized by the following steps: 1) Formula: [ka] Compound 37 of the formula: [ka] R 3 ' but independently C 1-6 To obtain compound 38, which is an alkyl group. 2) Compound 38 is reacted with the compound having the following structure: [ka] and reacting in the presence of a base with a compound 2a having the formula [ka] to form compound 39. 3) Reacting compound 39 with a first acid to give the following structure: [ka] to form compound 18a. 4) Compound 18a was dissolved in C 1-6 An alkyl alcohol and a second acid, and optionally a drying agent, are added and reacted to form compound 19.

[0019] In yet another aspect, the present invention relates to a compound of formula (III): [ka] [In the formula, R 1 is C 1-6 is alkyl; R 2 is C 1-3 is alkyl; R 4 is selected from NO2, N=O, NHOH, and NH2; and R 5 is selected from CHF2, CD3, and CH3. The present invention provides a compound of the formula:

[0020] In some embodiments of the compound of Formula (III), R 1 is methyl; R 2 is methyl; R 4 is selected from NO2 and NH2; and R 5 is CHF2.

[0021] In yet another aspect, the present invention relates to a compound of formula (IV): [ka] [In the formula, R 1 is C 1-6 is alkyl; R 2 is C 1-3 is alkyl; and R 5 is selected from CHF2, CD3, and CH3. The present invention provides a compound of the formula:

[0022] In some embodiments of the compound of Formula (IV), R 1 is methyl; R 2 is methyl; and R 5 is CHF2.

[0023] In yet another aspect, the present invention relates to a compound of formula (V): [ka] [In the formula, R 2 is C 1-3 is alkyl; and R 5 is selected from CHF2, CD3, and CH3. The present invention provides a compound of the formula:

[0024] In some embodiments of the compound of Formula (V), R 2 is methyl, and R 5 is CHF2.

[0025] In yet another aspect, the present invention relates to a compound of formula (VI): [ka] [In the formula, R2 is C 1-3 is alkyl; R 3 OH, O.C. 1-4 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3 selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I. or a free base form or salt thereof.

[0026] In some embodiments of the compound of Formula (VI), R 2 is C 1-3 is alkyl; R 3 OH, O.C. 1-4 Alkyl, [ka] Selected from; and X is selected from F, Cl, Br and I.

[0027] In some embodiments, the compound of Formula (VI) is compound 34 [ka] It has the structure:

[0028] In another aspect, the present invention provides a method for treating a thromboembolic disorder, comprising administering to a mammal, preferably a human, in need of such treatment a therapeutically effective amount of Compound (I), wherein Compound (I) is prepared using the novel synthesis method of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] definition As used herein, the term "alkyl" refers to a straight or branched chain saturated aliphatic group containing 1 to 10 carbon atoms, unless otherwise specified, and for example, alkyl includes methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, etc. The term "lower alkyl" refers to an alkyl group having 1 to 4 carbon atoms.

[0030] The term "alkoxy" refers to a group having the formula -O-alkyl, where an alkyl group, as defined above, is attached to the parent molecule through an oxygen atom. The alkyl portion of the alkoxy group can have from 1 to 10 carbon atoms (i.e., C1-C 10 alkoxy), or 1 to 6 carbon atoms (i.e., C1-C6 alkoxy). Examples of suitable alkoxy groups include, but are not limited to, methoxy (-O-CH3 or -OMe), ethoxy (-OCH2CH3 or -OEt), t-butoxy (-OC(CH3)3 or -OtBu), and the like.

[0031] The term "aryl" refers to a monocyclic or bicyclic fused ring containing 6 to 10 ring carbon atoms, where each ring is aromatic (eg, phenyl or naphthyl).

[0032] The term "substituents" refers to additional substituents selected from halogen (preferably fluoro, chloro, or bromo), hydroxy, amino, mercapto, etc. Preferred substituents in groups such as substituted lower alkyl or substituted alkyl described herein are halogen, especially fluoro groups.

[0033] The term "reducing agent" refers to any reagent that reduces the oxidation state of a carbon atom in a starting material by adding a hydrogen atom or an electron. These reagents will be apparent to one skilled in the art. Examples of "reducing agents" include, but are not limited to, borane-dimethylsulfide complex, 9-borabicyclo[3.3.1]nonane (9-BBN), catecholborane, lithium borohydride, sodium borohydride, sodium borohydride-methanol complex, potassium borohydride, sodium hydroxyborohydride, lithium triethylborohydride, lithium n-butylborohydride, sodium cyanoborohydride, calcium(II)borohydride, lithium aluminum hydride, diisobutylaluminum hydride, n-butyl-diisobutylaluminum hydride, sodium bis-methoxyethoxyaluminum hydride, triethoxysilane, diethoxymethylsilane, lithium hydride, lithium, sodium, Ni / B hydrogen, and the like. Certain acidic and Lewis acidic reagents enhance the activity of the reducing agent. Examples of such acidic reagents include acetic acid, methanesulfonic acid, and hydrochloric acid. Examples of such Lewis acidic reagents include trimethoxyborane, triethoxyborane, aluminum chloride, lithium chloride, vanadium(III) chloride, bis(cyclopentadienyl)titanium dichloride, cesium fluoride, potassium fluoride, zinc(II) chloride, zinc(II) bromide, and zinc(II) iodide.

[0034] The term "removable protecting group" or "protecting group" refers to any group that is attached to a functional site (e.g., the oxygen atom of a hydroxyl or carboxyl group, or the nitrogen atom of an amino group) to prevent reaction from occurring at that functional group, and that can be removed by a conventional chemical or enzymatic step to regenerate the original functional group. The type of removable protecting group used is not critical.

[0035] The term "ligand" as used herein refers to a phosphine derivative that binds to palladium, such as, for example, an arylphosphine or alkylphosphine that can form a complex with the palladium atom and is mono- or di-coordinated, as this term is well known to those skilled in the art in the particular field.

[0036] As used herein, the term "silylation" refers to the process of introducing silyl or silicon-containing groups, including, but not limited to, tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS), triethylsilyl (TES), trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl-oxy-methyl (TOM), and di-tert-butylsilyl bis(trifluoromethanesulfonate).

[0037] As used herein, the term "desilylation" refers to the process of removing a silyl or silicon-containing group.

[0038] EMBODIMENTS OF THE PRESENT DISCLOSURE The present invention is a number of synthetic intermediates and methods for the preparation of those intermediates and compounds (I).

[0039] General aspects of these example methods are illustrated in the Schemes and Examples. Each product used in the following processes is suitably separated, isolated, and / or purified before being used.

[0040] In general, the reaction conditions (e.g., temperature, reaction time, solvent, work-up method, etc.) followed for a particular reaction will be familiar to one skilled in the art. Typically, the temperature is between -100°C and 200°C, the solvent is aprotic or protic, and the reaction time is between 10 seconds and 10 days. Work-up typically consists of quenching any unreacted reagents, followed by partitioning (extraction) into an aqueous / organic layer, and separation of the layer containing the product.

[0041] Generally, the oxidation and reduction reactions are carried out at temperatures near room temperature (about 20°C), although metal hydride reductions often reduce the temperature to 0°C to -100°C. Generally, the solvent is aprotic for reduction reactions, and can be either protic or aprotic for oxidation reactions. Reaction times are adjusted to achieve the desired conversion.

[0042] In certain embodiments, the present invention provides a method for preparing Compound (I). Representative general methods for preparing derivatives are outlined in Schemes 1 and 2 below. [ka] [ka]

[0043] Each step of the preparation process shown in the above scheme is described in detail below.

[0044] Step 1

[0045] The starting materials for this process are compound 1 and compound 2. In embodiments in which the starting materials are prepared according to literature methods, the starting materials are preferably purified prior to reaction. Compounds 1 and 2 are reacted under basic conditions in a suitable solvent to form compound 3. Suitable bases include, for example, Li + , Na + , and K + as the counter cation (e.g., methoxide, ethoxide, tert-butoxide, amylate, tert-amylate).

[0046] Examples of suitable solvents include, but are not limited to, polar aprotic solvents (e.g., dimethylformamide, dimethylsulfoxide, and N-methylpyrrolidinone); ethereal solvents (e.g., tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), methyl t-butyl ether (MTBE), diethoxymethane, and CPME); hydrocarbon solvents (e.g., benzene, toluene, hexane, and heptane); halogenated solvents (e.g., dichloromethane and 1,2-dichloroethane); acetate solvents (e.g., ethyl acetate, isopropyl acetate, and butyl acetate), and other solvents (e.g., acetonitrile, methyl vinyl ketone, N,N-dimethylacetamide); polar aprotic solvents and mixtures thereof. Preferred solvents include ethereal solvents (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, and diethoxymethane).

[0047] This reaction can be carried out at about -78°C to about 0°C. The reaction is preferably carried out at about -50°C to about -20°C.

[0048] Step 2

[0049] Compound 3 is then subjected to a reverse Claisen reaction under acidic conditions or in an acidic aqueous solution to give compound 4. Suitable acids include, but are not limited to, formic acid, acetic acid, benzenesulfonic acid (BSA), nitric acid, perchloric acid, methanesulfonic acid (MSA), trifluoroacetic acid (TFA), citric acid, hydrochloric acid (HCl), sulfuric acid (H2SO4), and phosphoric acid (H3PO4). Preferably, the acid is MSA.

[0050] This reaction can be carried out over a relatively wide range of temperatures. The reaction is generally carried out at a temperature of 0° C. to 80° C. Preferably, the reaction is carried out at a temperature of about 20° C. to about 65° C.

[0051] Step 3

[0052] Compound 4 is then converted to the corresponding esters and ketals of compound 4 using an alcoholic solvent in the presence of a trialkyl orthoformate, an acid catalyst, and optionally a drying agent. In some embodiments, the alcoholic solvent is 1-6 The alcohol solvent (e.g., methanol, ethanol, propanol, butanol, pentanol, and hexanol), the acid catalyst may be selected from HCl, chlorotrimethylsilane (TMSCl), pyridinium p-toluenesulfonate (PPTS), p-toluenesulfonic acid (PTSA), if a drying agent is required it may be selected from Na2SO4 and MgSO4, and the trialkyl orthoformate may be selected from, but is not limited to, trimethyl orthoformate (TMOF) and triethyl orthoformate (TEOF).

[0053] Step 4

[0054] The ester of compound 5 is then hydrolyzed under basic conditions in the presence of water and a suitable organic solvent that is stable under basic conditions (e.g., toluene, NMP). + , Na + , K + , Cs + or NH4 + as the counter cation. Examples of hydroxides with associated counter cations include, but are not limited to, KOH, NaOH, and LiOH.

[0055] Step 5

[0056] The carboxylic acid of compound 6 can be further reacted with an activating agent to form an activated species that reacts directly with the chiral auxiliary to give compound 6a in the presence of a base. Typical activating agents include reagents such as acyl chlorides (e.g., pivaloyl chloride), isopropyl chloride, acid anhydrides (e.g., pivalic anhydride, isopropyl anhydride) or oxalyl chloride and sulfonyl chloride.

[0057] Chiral auxiliaries include, but are not limited to, oxazolidinone, 8-phenylmenthol, trans-phenylcyclohexanone, camphorsultam, pseudoephedrine (R,R) or (S,S), or pseudoephenamide (R,R) or (S,S), alkyl thiazolidine-2-thione derivatives, or N-(-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]heptan-2-yl)-N-phenylbenzenesulfonamide. In certain embodiments, the chiral auxiliaries are [ka] The oxazolidinone is selected from the group consisting of

[0058] The base may be selected from, for example, DIPEA, TEA, LDA, n-BuLi, sec-BuLi, or tert-BuLi, potassium tert-butoxide, with or without the presence of an inorganic base (eg, LiCl) in a suitable solvent.

[0059] Step 6

[0060] Compound 6a is alkylated with an alkylating agent and a strong base to give compound 6b. Examples of activating agents include, but are not limited to, alkyl halides, dialkyl sulfates, and trialkyloxonium tetrafluoroborate. Preferably, the alkylating agent is a methyl halide (e.g., MeI). Suitable bases are NaHMDS, LiHMDS, KHMDS, and LDA. Solvents can be selected from ethereal solvents (THF, 2-Me-THF, MTBE, CPME), aromatic solvents (toluene), or polar aprotic solvents, or combinations thereof. The chiral auxiliary is then removed under basic conditions using a hydroxide base (e.g., LiOH, NaOH, and KOH) to give compound 7.

[0061] Step 7

[0062] Compound 7 is isolated as an amine base or an alkali salt of Na or K in a desired solvent. Suitable bases are dibenzylamine, DABCO, dicyclohexylamine, ethanolamine, diethanolamine, imidazole, arginine, lysine, tromethamine, alanine, NaOH, KOH, LiOH. Suitable solvents are ether solvents (THF, 2-Me-THF, MTBE, CPME), aromatic solvents (toluene), ketone solvents (acetone, MIBK, MEK) or ester solvents (EtOAc, PrOAc), acetonitrile, and alcohol solvents (MeOH, EtOH, IPA). Alternatively, compound 7 may be isolated as a free acid.

[0063] Step 8

[0064] Compound 7 is then reacted with compound 8 in the presence of a metal catalyst and a base to give compound 9. The metal catalyst is derived from Pd, Pt, Rh, Ru, Ir, Fe, Ni or Cu. Ligands (e.g. phosphines, i.e. CX-A, XPhos, SPhos, Xantphos, DCEPhos) or N-heterocyclic carbenes (i.e. IMes, Ipr) can assist this reaction. Suitable bases include organic bases (i.e. Et3N, DIPEA), inorganic bases (i.e. KOPiv, KOAc, K2CO3), or bases derived from inorganic bases and carboxylic acids (i.e. K2CO3 / PivOH, Cs2CO3 / PivOH, K2CO3 / PhCO2H). Suitable solvents are ethereal solvents (i.e. THF, 2-Me-THF, MTBE, CPME), aromatic solvents (i.e. toluene, benzene), or polar aprotic solvents (i.e. DMF, DMAc, NMP).

[0065] Step 9

[0066] The nitro group of compound 9 is reduced using a metal catalyst (e.g., Pd, Pt, Rh on charcoal, aluminum oxide) in the presence of hydrogen gas or a hydrogen-displacing reagent (e.g., ammonium formate or sodium formate) in an ethereal or alcoholic solvent to form compound 10. Compound 9 can also be reacted with HSiCl3 / DIPEA, SnCl2, or Na2S2O4 to give compound 10.

[0067] Step 10

[0068] Compound 10 is then macrolactamized using a suitable carboxyl activating agent, base in a suitable solvent. Suitable coupling agents are any of the well-known coupling agents that couple amines with acids to form amides. Examples of coupling agents include, but are not limited to, PyBOP, HATU / HOBt, EDAC, oxalyl chloride, acid anhydrides (e.g., pivalic anhydride), acid chlorides (e.g., pivalic chloride), or activating agents (e.g., DPPCL, DMC, or TCFH). Suitable solvents typically include ethereal solvents (THF, 2-Me-THF, MTBE, CPME), aromatic solvents (toluene).

[0069] Step 11

[0070] In step 11, the ketone group of compound 11 is deprotected under aqueous acidic conditions to give compound 12. Examples of acids include, but are not limited to, HCl, HBr, and TFA.

[0071] Step 12

[0072] Compound 12 then undergoes a reductive amination step to give compound 13. This transformation can be carried out using a reducing agent (e.g., BH3, NaBH3CN, Pd / C, Pt / C) in the presence of an amine donor (e.g., an ammonium salt such as ammonia or ammonium chloride), a hydrogen replacement salt (e.g., ammonium formate, or hydrogen gas when Pd / C or Pt / C is used).

[0073] Reductive amination can also be achieved using aminotransferases in the presence of amine donors (e.g., isopropylamine, alanine, 3-aminobutyric acid, and methylbenzylamine) and cofactors (e.g., PLP). In the latter, the preferred solvent is aqueous DMSO. Different recyclization systems can be used (e.g., aminotransferase / lactate dehydrogenase / glucose dehydrogenase and aminotransferase / amino acid dehydrogenase / formate dehydrogenase). Examples of aminotransferases include, but are not limited to, ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260.

[0074] The aminotransferases used in the processes of the present disclosure generally comprise an amino acid sequence that is at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to a reference amino acid sequence selected from any one of ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260. In some embodiments, the aminotransferase is a recombinant aminotransferase polypeptide having an amino acid sequence that differs at one or more amino acid residues compared to a reference sequence (e.g., ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260). In some embodiments, a polynucleotide that is hybridizable under highly stringent conditions encodes an aminotransferase polypeptide that has the above-mentioned proportions of identical sequences and differs by one or more residues compared to a reference sequence (e.g., ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260).

[0075] In the processes described herein, the aminotransferase uses an amine donor to form a product compound. In some embodiments, the amine donor in the reaction conditions comprises a compound selected from isopropylamine (also referred to herein as "IPM") or any other suitable amine donor for the reaction of interest. In some embodiments, the amine donor is IPM.

[0076] Also, suitable reaction conditions for this process generally include cofactor in reaction mixture.Since aminotransferase generally uses a member of vitamin B6 family, reaction conditions include cofactor selected from pyridoxal-5'-phosphate (also known as pyridoxal phosphate, PLP, P5P), pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM), and their phosphorylation products; pyridoxine phosphate (PNP) and pyridoxamine phosphate (PMP).In some embodiments, suitable reaction conditions can include cofactor selected from PLP, PN, PL, PM, PNP, and PMP.In some embodiments, cofactor is PLP.

[0077] Step 13

[0078] Compound 13 is finally coupled with compound 14 to give compound (I) as shown in WO 2015 / 116886.

[0079] In the above process, other steps may be performed between steps 1-13. Additionally, different synthetic routes may be used to prepare key intermediates in Schemes 1 and 2. Scheme 3 shows an alternative method for preparing a specific example of compound 7 (Scheme 1) as compound 21. [ka]

[0080] Compound 15 can be coupled with 3-chloro-1,1-dimethoxypropane, 3-bromo-1,1-dimethoxypropane or 3-iodo-1,1-dimethoxypropane in the presence of a metal (e.g. Mg) and an initiator (e.g. I2) in a suitable solvent (e.g. THF) to form compound 16. Subsequent ketal hydrolysis utilizes an organic acid (e.g. TFA, MSA, BSA, PTSA, PPTS) or an inorganic acid (e.g. HCl, HBr) in the presence of water and a suitable solvent to give compound 17. The aldehyde of compound 17 is then reacted with a triphenylphosphonium ylide (e.g., methyl 2-(triphenylphosphoranylidene)propanoate or ethyl 2-(triphenylphosphoranylidene)propanoate) or with a phosphonate derivative (e.g., methyl 2-(diethoxyphosphoryl)propanoate or ethyl 2-(diethoxyphosphoryl)propanoate) in a suitable solvent in the presence of a base (e.g., NaH or KOtBu) to give compound 18. The ketone of compound 18 is then reacted with an alcoholic solvent (e.g., C 1-6 The ester is then protected using a 2-aminopropyl alcohol) and an acid as a catalyst (e.g., HCl, chlorotrimethylsilane (TMSCl), pyridinium p-toluenesulfonate (PPTS), p-toluenesulfonic acid (PTSA)) and an appropriate drying agent (e.g., Na2SO4, MgSO4, trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF)) to convert it to the corresponding ketal compound 19. The ester is then hydrolyzed under basic conditions in the presence of water to give compound 20. Suitable bases include Li + , Na + , K + , Cs + , NH4 + The hydroxide with 1 as the counter cation. Finally, the olefin of compound 20 is reduced using a metal catalyst in the presence of H2 to give compound 21. The metal is preferably Ru or Rh. Chirality at the methyl carbon center is introduced by the use of an appropriate chiral ligand. Alternatively, compound 19 can be reduced by treatment with an ene reductase enzyme and ester hydrolysis under basic conditions.

[0081] Compound 19 can be prepared by another condensation (Scheme 4). [ka]

[0082] Aldehyde 37 may be coupled with a triphenylphosphonium ylide (e.g., methyl 2-(triphenylphosphoranylidene)propanoate or ethyl 2-(triphenylphosphoranylidene)propanoate) to form compound 38, or may be reacted with a phosphonate derivative (e.g., methyl 2-(diethoxyphosphoryl)propanoate or ethyl 2-(diethoxyphosphoryl)propanoate) in a suitable solvent in the presence of a base (e.g., NaH or KOtBu) to give compound 38. The resulting bis-ester 38 is reacted with compound 2 having the structure of compound 2a in a suitable solvent in the presence of a base (e.g., LiHMDS, LDA, tBuOK) to give compound 39. This compound is further decarboxylated in a suitable solvent in the presence of an acid (e.g., HCl, MSA, H3PO4) to give compound 18a, which is subsequently converted to compound 19 as described above.

[0083] In another embodiment, compound 21 may be obtained starting from a cyclopentane ester derivative (Scheme 5). [ka]

[0084] The cyclopentane ester derivative is coupled with formaldehyde and a dialkylamine to form compound 40. Further treatment under basic conditions gives the diacid derivative, compound 41. After esterification to form compound 42, it is coupled with compound 2a and treated with acid to give the acrylic acid derivative, compound 18b. The ketone of compound 18b is then dissolved in an alcoholic solvent (e.g., C 1-6The ester is then protected using a 20-methyl-20-phenyl alcohol), an acid catalyst (e.g., HCl, chlorotrimethylsilane (TMSCl), pyridinium p-toluenesulfonate (PPTS), p-toluenesulfonic acid (PTSA)), and an appropriate drying agent (e.g., Na2SO4, MgSO4, trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF)) to give the corresponding ketal. The ester is then hydrolyzed under basic conditions in the presence of water to give compound 20b. Suitable bases include Li + , Na + , K + , Cs + , NH4 + The countercation includes the hydroxide or free carboxylic acid. Finally, the olefin of compound 20b is reduced in the presence of H2 using a metal catalyst to give compound 21. The metal is preferably Ru or Rh. Chirality at the methyl carbon center is introduced by the use of an appropriate chiral ligand. Alternatively, the desired enantiomer may be obtained by treatment with an ene reductase enzyme.

[0085] In another embodiment, specific examples compound 7 (Scheme 1), compound 27 (Scheme 7), which can be prepared by enzymatic resolution are shown in Reaction Schemes 6 and 7. [ka]

[0086] Compounds 22 and 2 are reacted in a suitable solvent under basic conditions to give compound 23. + , Na + , K +Bases such as alkoxides (methoxide, ethoxide, tert-butoxide, amylate, tert-amylate) with a countercation of are suitable for solvents such as ethereal solvents (THF, 2-MeTHF, MTBE, CPME), aromatic solvents (toluene) or dipolar aprotic solvents. The reverse Claisen reaction of compound 23 under acidic conditions or in aqueous acidic solution gives compound 24. Suitable acids include, but are not limited to, H2SO4, MSA, BSA, nitric acid, TFA or perchloric acid.

[0087] Compound 24 was then dissolved in an alcohol-based solvent (C 1-6 The corresponding esters and ketals of compounds 25 can be converted to the corresponding esters and ketals of compounds 26 using a 2-hydroxyethyl alcohol, and an acid catalyst (such as, but not limited to, HCl, chlorotrimethylsilane (TMSCl), pyridinium p-toluenesulfonate (PPTS), p-toluenesulfonic acid (PTSA)), and an appropriate drying agent (e.g., Na2SO4, MgSO4, and a trialkyl orthoformate (e.g., trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF)).

[0088] Compound 25 is then enzymatically resolved as shown in Scheme 7. [ka]

[0089] Racemic compound 25 is resolved enzymatically. The unwanted enantiomer in the racemic mixture of compound 25 is hydrolyzed while the desired enantiomer compound 26 remains unreacted. Compound 26 is then hydrolyzed using aqueous basic solution (e.g. NaOH) to give compound 27. Preferably, the hydrolytic enzyme is lipase MH Amano 10 SD, which exhibits high selectivity (enantiomeric excess >90%).

[0090] In method B, racemic compound 25 is resolved enzymatically. The desired enantiomer in the racemic mixture compound 25 is hydrolyzed to give compound 27, while the undesired enantiomer remains partially unreacted.

[0091] In Method C, the unreacted unwanted enantiomer compound S-26 produced in Method B is racemized in the presence of base to form compound 25 which is used as the starting material in Method A or Method B above.

[0092] In an alternative embodiment, intermediate compound 10 is prepared by a method different from Scheme 1, as shown in Scheme 8. [ka]

[0093] Step 1

[0094] Compound 28 is coupled with a silyl-protected acetylene in a strong base and a suitable solvent to give compound 29. The base may be a strong lithium base, such as an alkyl or aryl lithium base. Examples of alkyl and aryl lithium bases include, but are not limited to, methyl lithium, n-butyl lithium, sec-butyl lithium, tert-butyl lithium, and phenyl lithium. The solvent may be an ethereal solvent, such as THF.

[0095] Step 2 The ketone moiety is then reacted with an alcohol-based solvent (C 1-6 The silyl groups are then protected as the corresponding ketals using a fluoride source (e.g., TBAF, HF TEA, HF) in an appropriate solvent (e.g., THF, 2-MeTHF) to give compound 30.

[0096] Step 3

[0097] The triple bond of compound 30 is then derivatized to the corresponding vinyl halide compound 31 in two steps using LiAl(OtBu)3H / Cp2ZrCl2 followed by a halide donor such as N-chlorosuccinimide, N-bromosuccinimide or N-iodosuccinimide.

[0098] Step 4

[0099] Compound 31 is then metal-catalyzed cross-coupled with commercially available (S)-(-)-3-methoxy-2-methyl-3-oxopropylzinc bromide to give compound 32. Examples of metal catalysts include, but are not limited to, Pd(II) salts (e.g., PdCl2, Pd(OAc)2), or coordination metals (e.g., 1,1'-bis(di-tert-butylphosphino)ferrocenedichloropalladium).

[0100] Step 5

[0101] Compound 32 is then reacted with compound 8 in the presence of a metal catalyst and a base to give compound 33. The metal catalyst can be derived from Pd, Pt, Rh, Ru, Ir, Fe, Ni or Cu. Ligands (e.g., phosphines, i.e., CX-A, XPhos, SPhos, Xantphos, DCEPhos) or N-heterocyclic carbenes (i.e., IMes, Ipr) can aid in the reaction. Suitable bases include organic bases (i.e., Et3N, DIPEA), inorganic bases (i.e., KOPiv, KOAc, K2CO3), or bases derived from inorganic bases and carboxylic acids (i.e., K2CO3 / PivOH, Cs2CO3 / PivOH, K2CO3 / PhCO2H). Suitable solvents are ethereal solvents (i.e., THF, 2-Me-THF, MTBE, CPME), aromatic solvents (i.e., toluene, benzene), or polar aprotic solvents (i.e., DMF, DMAc, NMP).

[0102] Step 6

[0103] Compound 33 is then subjected to reducing conditions to reduce the double bond and nitro group, followed by hydrolysis of the methyl ester to give compound 10. Reduction is effected using a metal (e.g., Pd or Pt) in the presence of hydrogen gas in a protic solvent (e.g., MeOH, EtOH, IPA). Ester hydrolysis occurs by treating the methyl ester with a hydroxide base (e.g., LiOH, NaOH, KOH) in the presence of water or water and a miscible organic solvent. [ka]

[0104] Step 1

[0105] Compound 45 can be synthesized from compounds 43 and 44 under suitable Suzuki coupling conditions, for example, in a suitable solvent (e.g., methanol, DMF, or acetonitrile) in the presence of an appropriate level of a palladium catalyst (e.g., Pd(PPh3)4, Pd(OAc)2, or Pd(dppf)Cl2-DCM complex).

[0106] Steps 2 and 3

[0107] Compound 46 can be obtained by azidation followed by click reaction with an appropriate acetylene compound. Compound 45 is treated with azidation conditions (e.g., TMSN3 / tBuONO) to give an intermediate azide, which is then reacted with trimethylsilylacetylene in the presence of a copper(I) catalyst (e.g., CuOAc or copper(I) iodide) to give triazole compound 46.

[0108] Step 4

[0109] Compound 47 can be obtained by reacting silyl compound 46 with 1,3-dichloro-5,5-dimethylhydantoin in a suitable solvent, including polar aprotic solvents such as THF or DMF.

[0110] Step 5

[0111] Compound 14 can be obtained by reacting compound 47 in hydrochloric acid (eg, concentrated hydrochloric acid).

[0112] In another embodiment, the present invention relates to a compound of formula (II): [ka] [In the formula, The dashed lines (----) are optional bonds; R 1 is C 1-6 is alkyl; R 2 is C 1-3 Alkyl, alkenyl; R 3 OH, O.C. 1-6 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3 selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I. The compound of formula (I) is provided in base or salt form.

[0113] In another embodiment, the present invention provides [ka] The present invention provides a compound selected from the group consisting of:

[0114] In another embodiment, the present invention provides [ka] The present invention provides a compound having a structure selected from the group consisting of:

[0115] In another embodiment, the present invention provides [ka] The present invention provides a compound selected from the group consisting of:

[0116] In another embodiment, the present invention relates to a compound of formula (III): [ka] [In the formula, R 1 is C 1-6 is alkyl; R 2 is C 1-3 is alkyl; R 4 is selected from NO2, N=O, NHOH, and NH2; and R 5 is selected from CHF2, CD3, and CH3. The present invention provides a compound of the formula:

[0117] In another embodiment, the present invention provides [ka] The present invention provides a compound selected from the group consisting of:

[0118] In another embodiment, the present invention relates to a compound of formula (IV): [ka] [In the formula, R 1 is C 1-6 is alkyl; R 2 is C 1-3 is alkyl; and R 5 is selected from CHF2, CD3, and CH3. The present invention provides a compound of the formula:

[0119] In another embodiment, the present invention provides [ka] The present invention provides a compound selected from the group consisting of:

[0120] In another embodiment, the present invention relates to a compound of formula (V): [ka] [In the formula, R 2 is C 1-3 is alkyl; and R 5 is selected from CHF2, CD3, and CH3." The present invention provides a compound of the formula:

[0121] In another embodiment, the present invention provides [ka] The present invention provides a compound of the formula:

[0122] In another embodiment, the present invention relates to a compound of formula (VI): [ka] [In the formula, R 2 is C 1-3 is alkyl; R 3 OH, O.C. 1-6 Alkyl, [ka] Selected from; R 6 is C 1-3 selected from alkyl, phenyl, and benzyl; R 7 is selected from H and phenyl; R 8 is C 1-3 selected from alkyl, phenyl, and benzyl; and X is selected from F, Cl, Br and I. The present invention provides a compound of the formula:

[0123] In another embodiment, the present invention provides a compound having the structure: [ka] The present invention provides a compound having the formula: EXAMPLES

[0124] To better understand the present invention, the following examples are provided. All reactions were carried out under anhydrous conditions under nitrogen atmosphere unless otherwise noted. Reagents were used as received from the vendor unless otherwise noted. Yields reported are for isolated materials and have not been corrected for water content. Reactions were monitored by normal or reversed phase HPLC systems (Shimadzu) using CH3CN / H2O / MeOH (containing either 0.05% TFA or 0.1% NHOAc) as the mobile phase.

[0125] Method A

[0126] Chromatography conditions [Table 1]

[0127] Gradient [Table 2]

[0128] Method B

[0129] Chromatography conditions [Table 3]

[0130] Gradient [Table 4]

[0131] Method C

[0132] Chromatography conditions [Table 5]

[0133] Gradient [Table 6]

[0134] Method D

[0135] Chromatography conditions [Table 7]

[0136] Gradient [Table 8]

[0137] Method E

[0138] Chromatography conditions [Table 9]

[0139] Gradient [Table 10]

[0140] Method F

[0141] Chromatography conditions [Table 11]

[0142] Gradient [Table 12]

[0143] Method G

[0144] Chromatography conditions [Table 13]

[0145] Gradient [Table 14]

[0146] Method H

[0147] Chromatography conditions [Table 15]

[0148] Gradient [Table 16]

[0149] Method I

[0150] Chromatography conditions [Table 17]

[0151] Gradient: [Table 18]

[0152] Method J

[0153] Chromatography conditions [Table 19]

[0154] Gradient [Table 20]

[0155] Method K

[0156] Chromatography conditions [Table 21]

[0157] Gradient [Table 22]

[0158] Method L

[0159] Chromatography conditions [Table 23]

[0160] Gradient [Table 24]

[0161] NMR spectra were recorded on a Bruker DRX-600, DRX-500 or DRX 400 and were referenced to residual protons of deuterated solvents. Low resolution mass spectra (LRMS) were recorded on a Waters ZQ ES.

[0162] The abbreviations used herein are defined as follows: "1x" is one time, "2x" is two times, "3x" is three times, "°C" is degrees Celsius, "eq" is equivalent, "g" is gram, "mg" is milligram, "L" is liter, "mL" is milliliter, "μL" is microliter, "N" is normality, "M" is molar, "mmol" is millimole, "min" is minute, "h" is hour, "rt" is room temperature, "RT" is retention time, "atm" is atmospheric pressure, "psi" is pounds per square inch, "conc." is concentration, "sat" or "sat'd" is saturation, "MW" is molecular weight, "mp" is melting point, "ee" is enantiomeric excess, "MS" or "Mass Spec" is mass spectrometry, "ESI" is electrospray ionization mass spectrometry, "HR" is high resolution, "HRMS" is high resolution mass spectrometry, "LCMS" is liquid chromatography mass spectrometer, "HPLC" is high performance liquid chromatography, "RPMS" is HPLC ... "HPLC" stands for reverse phase HPLC, "TLC or "tlc" stands for thin layer chromatography, "NMR" stands for nuclear magnetic resonance spectroscopy, "nOe" stands for nuclear Overhauser effect spectroscopy, 1 "H" is proton, "δ" is delta, "s" is singlet, "d" is doublet, "t" is triplet, "q" is quartet, "m" is multiplet, "br" is broad, "Hz" is Hertz, and "α", "β", "R", "S", "E", and "Z" are symbols used in stereochemistry that are familiar to those of skill in the art. [Table 25] [Table 26] [Table 27]

[0163] The preparation of intermediate compounds 3a-Cl, 4a, 5a, 6a, 7a, 35, and 36 is described in Scheme 10 (an embodiment of general Scheme 1 described above) and Examples 1-4. An embodiment of general Scheme 2 for forming compound (I) is described in Examples 5-12 below. [ka] Example 1 Synthesis of compound 3a-Cl [ka]

[0164] To a 20 L reaction vessel equipped with a nitrogen inlet, temperature probe, and overhead stirrer, compound 2a (540 g, 3053 mmol, limiting reagent) was added followed by THF (4500 mL) and stirred to dissolve compound 2a. Compound 1 (287.4 g, 3382 mmol, 1.2 equiv) was then added followed by a THF (50 mL) rinse.

[0165] The solution was cooled with a refrigerator to −35° C. Then potassium tert-butoxide / THF solution (1 M, 3650 mL, 1.3 equiv.) was added slowly so that the temperature did not exceed −30° C.

[0166] The reaction was held at -35 to -30 °C for 1 h until the reaction was complete. To a separate 20 L reaction vessel, water (3635 mL) was added followed by concentrated sulfuric acid (193.3 g, 0.69 equiv). The vessel temperature of the reaction vessel was set to 10 °C and the reaction was cooled to 12 °C. The cold solution from the aforementioned cold vessel (-35 to -30 °C) was added to the 20 L reaction vessel containing the cold aqueous H2SO4 solution using a transfer tube while maintaining the temperature below 10 °C. After the transfer was complete, THF was then distilled under vacuum at 20 to 25 °C until the volume was ~7.5 L. At this point a solid formed. The slurry was filtered and washed with water (2000 mL, 3.7 V). Compound 3a-Cl [1,061 g) was obtained as a tan solid.

[0167] 1H NMR(500MHz, DMSO-d6)δ 14.97 (br s, 0.5H), 8.73 (t, J=5.1Hz, 1H), 8.02-7.98 (d, J= 24.1Hz, 1H), 7.85-7.75 (dd, J= 32, 5.1Hz, 1H), 4.70 (t, J=9.5Hz, 0.5H), 2.93 (t, J=7.2Hz, 1H), 2.61-2.53 (m, 1H), 2.44-2.21 (m, 2H), 2.08 (ddd, J=12.3, 8.2, 4.0Hz, 1H), 2.01-1.81 (m, 1H)

[0168] C 11 H 11 ClNO2 + LRMS calculated value: [M+H] + 224.05, observed: 224.28

[0169] Example 2 Synthesis of compound 4a To a 20 L cold vessel was added 70 wt% MSA solution (1074 g, 7823 mmol, 2.83 equiv.) followed by water (4900 mL). The reaction mixture was heated at 65 °C. The vessel was then added with a wet cake of compound 3a-Cl and heated at 65 °C for 3 h until the reaction was complete. The reaction was then cooled to 20-25 °C and 28 wt% aqueous NH4OH solution (489 g, 3907 mmol, 1.41 equiv.) was added. The pH was adjusted to 5.06 using a pH probe. The resulting slurry was heated at 44 °C and held at 44 °C overnight. The reaction was cooled to 20-25 °C, the slurry was filtered, and the resulting cake was washed with water (3000 mL, 6 V). The wet cake was dried in a vacuum oven (50 °C, 100 mmHg) for 2 days to give compound 4a (590 g) as a white solid.

[0170] 1H NMR (500MHz, DMSO-d6): δ 12.01 (br s, 1H), 8.75-8.67 (m, 1H), 7.94 (br s, 1H), 7.86-7.78 (m, 1H), 3.16 (br t, J=6.8Hz, 2H), 2.25 (br t, J=6.6Hz, 2H), 1.70-1.60 (m, 2H), 1.60-1.50 (m, 2H)

[0171] C 11 H 13 ClNO3 + LRMS calculated value: [M+H] + 242.06, Observed: 242.24

[0172] Example 3 Synthesis of compounds 5a and 6-Cl TIFF0007631218000108.tif43164To a 20 L reaction vessel was added MeOH (4 L) and compound 4a (367.3 g, 1500 mmol, 98.9 wt%). This was followed by the addition of 3 L of MeOH. Trimethyl orthoformate (734 mL, 6700 mmol, 100 wt%, 4.5 equiv) was then added, followed by a MeOH (400 mL) rinse. Chlorotrimethylsilane (367 mL, 2880 mmol, 100 wt%, 1.92 equiv) was added, followed by MeOH (200 mL). The resulting reaction was heated at an internal temperature of 49° C. for 12 hours.

[0173] To a separate 20 L reaction vessel, NaOH (10N, 1220 mL) was added followed by H2O (1620 mL) and the resulting reaction was cooled to 0°C. The reaction containing compound 5a from above was transferred to the reaction containing aqueous NaOH. The internal temperature rose to 5°C-22°C. MeOH (700 mL) was used to rinse the reaction vessel and the solution was transferred to the quench vessel. The reaction was stirred for 4 hours. The vessel temperature of the reaction vessel was warmed to 20°C and stirred. MTBE (2570 mL) was then added. Stirring was stopped and the product rich aqueous layer was collected and used next. 20 wt% citric acid (2985 mL) was then added to the stirred aqueous layer. Upon reaching pH 5.3, a slurry formed which was filtered to give compound 6-Cl as a solid (387.1 g, 89.6% yield).

[0174] 1 H NMR(400MHz, DMSO-d6)δ 11.98 (br s, 1H), (8.61 (d, J=4.8Hz, 1H), 7.60 (d, J=1.8Hz, 1H), 7.51 (dd, J=5.2, 2.1Hz, 1H), 3.03 (s, 6H), 2.08 (t, J=7.5Hz, 2H), 2.02 (br d, J=16.9Hz, 2H), 1.42-1.30 (m, 2H), 0.90-0.78 (m, 2H)

[0175] C 12 H 15 ClNO3 + LRMS calculated value: [M-CH3O] + 256.07, Observed: 256.24 Example 4 Synthesis of compound 7a-DCHA [ka]

[0176] Compound 6a (87.88 g, 305 mmol, limiting reagent) was added to a reaction vessel (2 L, chem-glass) followed by anhydrous THF (1760 mL). THF was distilled to 10 Vol. The KF of the solution was less than 200 ppm. Further THF (880 mL) was added along with triethylamine (CAS: 121-44-8, 106.4 mL, 2.5 equiv.). The solution was cooled to 0° C. and pivaloyl chloride (CAS: 3282-30-2, 44.13 g, 1.2 equiv.) was added via addition funnel at such a rate that the temperature did not exceed 5° C. After 30 min, lithium chloride (16.16 g, 1.2 equiv.) was added. After aging for 15 min, the chiral auxiliary (CAS: 102049-44-7, 64.86 g, 1.2 equiv.) was added as a solid in one portion. The slurry was allowed to warm to 20°C over 3 hours and left to stand overnight. The THF was then distilled under reduced pressure to a final volume of 800 mL. Toluene (530 mL) was added followed by saturated aqueous NH4Cl (270 mL) and water (270 mL). After mixing for 15 minutes the two phases were separated and the lower aqueous layer was removed. The organic layer was washed with 7 wt% NaHCO3 (270 mL) and water (270 mL). After phase separation the lower aqueous layer was removed. The organic layer was distilled to 220 mL. Anhydrous THF (1860 mL) was then added. The solution was passed through 0.45 micron polish and filtered.

[0177] The solution containing compound 35 was then cooled to -45°C and methyl iodide (95.4g, 2.2eq) was added followed by 1N NaHMDS / THF (458mL, 1.5eq) at such a rate that the temperature did not exceed -39°C. The reaction was allowed to continue for 6 hours. The resulting reaction mixture was then neutralized in one portion with a solution of acetic acid (29.30g, 1.6eq) / anhydrous THF (88mL). The organic reaction solution was washed with a 14wt% NaCl solution (530mL) followed by a 7.0wt% NaHCO3 solution (530mL). After washing, the organic solution containing compound 36 was concentrated to 220mL.

[0178] THF (880 mL) was added and the solution was cooled to 0° C. A 30 wt % H2O2 solution (64.18 g, 1.82 equiv.) was then added, followed by lithium hydroxide solution (12.42 g / 110 mL water) over 10 min. After 6 h reaction, a 10 wt % sodium bisulfite solution (63.48 g, 2.0 equiv. / 580 mL water) was added. The mixture was aged for 1 h. The THF was then distilled off to ∼700 mL, which was collected.

[0179] The pH was then adjusted to about 9.5 with NaOH (10N). Toluene (540 mL) was added. The two-phase mixture was mixed for 15 minutes and then allowed to settle. The separated organic layer was further extracted with saturated NaHCO3 (360 mL). The combined aqueous layers were returned to the reaction vessel and extracted with MTBE (720 mL). The organic layer was removed.

[0180] The product-rich aqueous layer containing compound 7a was returned to the reaction vessel and MTBE (900 mL) was added. The pH was adjusted to 4.4 with citric acid.

[0181] MTBE was removed by distillation and replaced with MeCN / MTBE (4:1, 6 volumes) based on the amount of compound 7a added (corrected effective amount). The resulting solution was polish filtered. Dicyclohexylamine (up to 1.5 equivalents relative to compound 7a) was then added in one portion. The slurry was heated at 55° C. and allowed to react for 30 minutes. The reaction was then cooled to 0° C. The slurry was filtered using a Buchner funnel under N2 protection, washed with cold MeCN (2.0 volumes) at 0° C., vacuum dried, and then dried in a vacuum oven at 50° C. for 24 hours. Compound 7a-DCHA was obtained as a white solid, 61 g (88.2% yield for the salt formation step, 61.6% yield over all four steps from compound 6a). Compound 7 is a 1:1.5 complex of compound 7:dicyclohexylamine.

[0182] 1H NMR (400MHz, MeOH-d4): 8.52 (d, J=5.3Hz, 1H), 7.72 (d, J=2.0Hz, 1H), 7.44 (dd, J=5.3, 2.0Hz, 1H), 3.16 (d, J=3.0Hz, 6H), 3.07-2.97 (m, 3H), 2.20-2.06 (m, 3H), 2.05-1.99 (m, 6H), 1.90-1.80 (m, 6H), 1.76-1.67 (m, 3H), 1.60-1.47 (m, 1H), 1.33-1.10 (m, 16H), 1.00 (d, J=6.8Hz, 3H), 0.97-0.87 (m, 2H)

[0183] C 13 H 17 ClNO3 + LRMS calculated value: [M-CH3O] + 270.09, Observed: 270.24 Example 5 Synthesis of compound 9a [ka]

[0184] To a clean 2 L reaction vessel equipped with an overhead stirrer, temperature probe, and nitrogen inlet, 2MeTHF (500 mL), catalyst [Pd(allyl)Cl]2 (1.57 g, 0.05 equiv.) and Xphos (4.48 g, 0.055 equiv.) were added in sequence at 20 °C to give a nearly homogeneous pale yellow solution. The salt compound 7a-DCHA (98.2 g, 1.0 equiv.) and pyrrole compound 8 (32.3 g, 1.15 equiv.) were added in portions to the reaction vessel to give a white suspension. After 30 min, KOPiv (32.3 g, 1.3 equiv.) was added to the solution in one portion and the reaction vessel was rinsed with 2-MeTHF (500 mL) that had been bubbled with N2 for 30 min. The solution was refluxed under N2 for 10 h to give a black suspension. The resulting crude was cooled to 20°C and quenched with K3PO4 (550mL, 20% aqueous solution) between pH 10.0-10.5, the aqueous layer was separated and the organic layer was washed with K3PO4-K2HPO4 buffer solution (800mL, pH 10.2 aqueous solution). The aqueous layers were combined and filtered to give a dark solution. To the aqueous solution, 2-MeTHF (1300mL) and activated charcoal (13.9g, Darco G-60) were added and the solution was acidified to pH 5-6 by the addition of citric acid (254g, 3.4eq) in portions over 30 minutes. The suspension was stirred for 30 minutes at 20°C. The suspension was filtered and the organic layer was concentrated in vacuuo to 300mL and solvent exchanged to nBuOH (1000mL) under reduced pressure (150mbar) at 80°C. The concentration of the resulting solution was adjusted to 170-180 mg / mL (5% by weight or less of 2-MeTHF in 500 mL of solution). The solution was gradually cooled to 0°C over 10 h and kept at 0°C for an additional 10 h to obtain a white slurry. The slurry was filtered through a Nutsche filter, the reaction vessel was rinsed with nBuOH (100 mL), and a cake was obtained from the resulting suspension. The cake was rinsed with heptane (100 mL) and dried in an oven (vacuum, 50°C for 24 h). The isolated compound 9a (73.2 g, 95% by weight) was obtained in 83% yield.

[0185] 1H NMR (400MHz, DMSO-d6): δ 8.89-8.80 (m, 1H), 8.72-8.65 (m, 1H), 7.88-7.52 (m, 3H), 3.10-3.00 (m, 6H), 2.50 (dt, J=3.5, 1.8Hz, 1H), 2.52-2.44 (m, 1H), 2.22-2.13 (m, 1H), 2.11-1.99 (m, 2H), 1.51-1.35 (m, 1H), 1.27-1.14 (m, 1H), 1.00-0.84 (m, 5H). 13 C NMR (101MHz, DMSO-d6) δ 177.3, 158.9, 149.5, 139.3, 138.3, 135.0, 132.8, 123.5, 123.0, 110.5 (t, J=253.9Hz, 1C), 103.0, 48.2, 48.2, 38.4, 34.0, 32.9, 20.4, 16.6

[0186] LRMS [M-OMe] + C 17 H 19 F2N4O5 + : 397.36, 397.13 Example 6 Synthesis of compound 10a [ka]

[0187] THF (900 mL), Pd / C (4.6 g, 10 wt%, 0.1 equiv), and compound 9a (46.0 g, 1.0 equiv) were added in sequence to a pressure-resistant reaction vessel equipped with an overhead stirrer, temperature probe, and nitrogen inlet at 20 °C to give a suspension. The reaction vessel was purged with N2 and H2 three times each. The solution was stirred vigorously under H2 (40 psi) for 18 h. The resulting crude was removed from the reaction vessel and filtered through a Nutsche. The THF solution was concentrated to a clear oil to give compound 10a (101 g, 40 wt%) in 95% yield. A small sample was removed, concentrated sufficiently, and used for spectral analysis.

[0188] 1 H NMR (500MHz, CDCl3): δ 8.77 (1H, s, br), 7.75 (1H, s. br), 7.39 (1H, s. br), 7.33 (1H, s. br), 7.11 (1H, t, J=59.1Hz), 5.75 (3H, s, br), 3.17 (6H, s), 2.35-2.25 (1H, m), 2.20-1.98 (2H, m), 1.55-1.43 (1H, m), 1.30-1.13 (1H, m), 0.80-1.09 (5H, m)

[0189] LRMS [M+H] + C 18 H 25 F2N4O4 + : 399.18 Example 7 Synthesis of compound 11a-FUM [ka]

[0190] To a 1 L reaction vessel, THF (600 mL) was added followed by chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH, 17.1 g, 59.7 mmol, 98.0 wt%, 1.52 equiv). Further THF (200 mL) was used to rinse in order to add all the TCFH to the reaction vessel. To this suspension, N,N-diisopropylethylamine (14.6 mL, 83.7 mmol, 100 wt%, 2.13 equiv) was added and stirred at room temperature. To a 50 mL syringe, a solution of compound 10a in THF (44.9 g, 39.2 mmol, 34.8 wt%, 1.00 equiv) was added to the reaction vessel by syringe over 10 h (rate: approx. 5 mL / h).

[0191] The resulting reaction mixture (~50 mL per 1 g of compound 11a added) was solvent exchanged from THF to MIBK (~20 mL). The organic layer was washed with aqueous K2HPO4 (15%, 15 mL) followed by the addition of 1.2 equivalents (per mole of compound 11a added) of solid fumaric acid. The mixture was then concentrated under reduced pressure to ~8 mL. A tan slurry product was formed, which was filtered and the cake washed with MIBK (2 mL) followed by heptane (2 mL) and dried under vacuum. Yield=1.01 g (75.3% of corrected effective amount of compound 11a added); effective amount=74.2 wt%; ee=97.3%

[0192] 1 H NMR(400MHz, DMSO-d6)δ 13.31-13.02 (m, 1H), 9.30 (s, 1H), 8.76 (d, J=5.1Hz, 1H), 7.94 (s, 1H), 7.87-7.79 (m, 1H), 7.66-7.60 (m, 1H), 7.41-7.35 (m, 2H), 6.64 (s, 2H), 3.22 (s, 3H), 3.15 (s, 3H), 2.41-2.28 (m, 1H), 1.85-1.60 (m, 3H), 1.55-1.41 (m, 1H), 0.88 (br d, J=7.1Hz, 4H), 0.46 (br s, 1H)

[0193] C 17 H 19 F2N4O2 + LRMS calculated value: 349.15 [M-CH3O] + , observed value: 349.08 Example 8 Synthesis of compound 12 [ka]

[0194] To a slurry of compound 11a-FUM (0.50 g, 67 wt%, 96 ee%) in water (5 mL) was added cyclopentyl methyl ether (2.5 mL) followed by trifluoroacetic acid (0.23 mL, 3.5 equiv). The resulting mixture was heated at 45° C. for 5 h. The mixture was then cooled to ambient temperature and filtered. The reaction vessel was rinsed with water (2.5 mL), which was used to wash the filter cake. The filtrates were combined and the phases were separated. The resulting organic layer was extracted with aqueous HCl (0.5 N, 2.0 mL). The extracted acidic aqueous solution was combined with the acidic aqueous layer obtained from the reaction. The pH of the combined aqueous solution was adjusted to 9-10 by adding solid K3PO4 (~2 g). The resulting mixture was stirred for 2 h and filtered. The filtered cake was washed with water (5 mL x 2) and MTBE (5 mL x 2) and dried under reduced pressure to give compound 12 (0.25 g, 78%, 97.2 ee%).

[0195] 1 H NMR (500MHz, CDCl3): δ 8.89 (s, 1H), 7.97 (s, 1H), 7.68 (s, 2H), 7.46 (s, 1H), 7.34 (t, J=59.7Hz, 1H), 6.85 (br s, 1H), 3.14 (br s, 1H), 2.82 (br d, J=8.9Hz, 1H), 2.46-2.37 (m, 1H), 2.05 (br s, 1H), 1.68 (br d, J=7.0Hz, 1H), 1.41 (br s, 1H), 1.21 (br s, 3H)

[0196] LRMS [M+H] + C 16 H 17 F2N4O2 + : 335.18 Example 9 Alternative synthesis of compound 12 [ka]

[0197] To a pressure vessel (1 L) was added compound 9a (75.0 g, 169 mmol), THF (525 mL), and Pt / V / C (~50% wet, 10 wt%, 7.5 g). The reaction vessel was flushed with N2 and H2 three times each. The solution was vigorously stirred at 20 °C for 1 h and heated at 40 °C for 16 h under H2 (1.5 bar). The mixture was removed from the reaction vessel, filtered, and the resulting solution was concentrated and the THF was distilled, followed by azeotropic distillation of water. To a separate reaction vessel was added THF (1.2 L), followed by chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH, 70 g, 1.52 equiv.) and N,N-diisopropylamine (103 mL, 3.5 equiv.). The mixture was vigorously stirred and heated at 55 °C. To this mixture was added a solution of compound 10a / THF over 20 hours (~50 mL / hr). The mixture was then concentrated and solvent exchanged to acetonitrile to give ~4 L / kg compound 11a / acetonitrile. Aqueous hydrochloric acid (3N, 23 mL) was then added and the resulting reaction mixture was heated at 55°C for 15 hours to give crude compound 12. The reaction was cooled to 10°C and dichloromethane (270 mL), water (540 mL) and hydrochloric acid (10.8N, 71 mL) were added. After stirring for 1 hour, the layers were separated and the lower organic layer was removed. To the aqueous layer was added potassium hydroxide (22.5% by weight, ~169 mL) at 10°C to adjust the pH to 3.0. After stirring at pH 3 for 1 hour, additional potassium hydroxide (22.5% by weight, ~17 mL) was added until the reaction was adjusted to pH 9.5. Compound 12 was isolated by filtration, rinsed with water (169 mL) and EtOH (114 mL), and then dried. Example 10 Synthesis of compound 13 [ka]

[0198] A reaction vessel (125 mL) equipped with a water circulator was used to maintain the reaction temperature at 35°C throughout the reaction. A calibrated pH meter was used to control the pH to 7.5 when 4 M aqueous isopropylamine was added to the reaction vessel. Ketone compound 12 (5.0 g, 50 g / L), DMSO (30 mL, 30%), and pyridoxal-5'-phosphate monohydrate (53 mg, final concentration 2.0 mM) were added to the reaction vessel. 1 M aqueous isopropylamine hydrochloride solution was prepared and 63 mL was added to the reaction vessel (final concentration 0.7 M). The reaction mixture was stirred for 2.0 minutes. The resulting reaction solution was heterogeneous. Aminotransferase: ATA-237 (0.5 g) was dissolved in 1 M isopropylamine hydrochloride solution (4.0 mL) and added to the reaction vessel. The enzyme vessel was rinsed with 1 M isopropylamine hydrochloride solution (3.0 mL) and added to the same reaction vessel. Samples (20 μL) were pipetted, diluted with methanol (980 μL), vortexed, centrifuged at 14000 x g for 2.0 min, filtered through a PTFE filter (0.2 μM), and analyzed for conversion and ee by HPLC. The reaction was stopped after 8 h (99.7% conversion).

[0199] The resulting reaction mixture was acidified to pH 1.3 (6N HCl, 3.6 mL). A celite pad was made and the reaction mixture was filtered through the celite. After filtration, the reaction vessel and celite pad were rinsed with water (30 mL) and the solution was pooled with the filtrate. The reaction mixture was extracted with 2-methyltetrahydrofuran (130 mL) and the 2-methyltetrahydrofuran solution was removed. The pH of the aqueous layer was adjusted to 10.5 with 10N sodium hydroxide (4.4 mL). The aqueous layer (152 mL) was extracted with n-butanol (150 mL) and the layers were separated (organic layer volume: 190 mL, aqueous layer volume: 105 mL). The aqueous layer (105 mL) was extracted again with n-butanol (100 mL) and the aqueous and organic layers were separated (aqueous layer volume: 75 mL, organic layer volume: 125 mL).

[0200] The organic layer was collected and the solvent was concentrated to 40 g of a viscous liquid. The resulting residue solidified at 4 °C for 1 h. The residue was suspended in MTBE (240 mL) and stirred vigorously. The precipitate was filtered and the filtrate was removed. The precipitate (6.6 g) was stirred with water (50 mL) and the pH was adjusted to 8.5. The desired compound precipitated, which was filtered and washed with MTBE. The volume of the filtrate was reduced (15 mL), the pH was adjusted to 9.0, and the precipitated compound was filtered and washed with MTBE. The two portions were combined and dried overnight in a vacuum oven at 35 °C. The desired compound 13 was isolated as an off-white solid (3.94 g, 78.8% yield, AP 99.3, ee>99.9%, effective amount 97%).

[0201] 1 H NMR (400MHz, DMSO-d6)δ 9.33 (s, 1H), 8.76-8.62 (m, 1H), 7.94 (t, J H-F =57.8Hz, 1H), 7.84 (s, 1H), 7.42-7.33 (m, 1H), 7.33-7.24 (m, 1H), 4.02-3.86 (m, 1H), 2.61-2.52 (m, 1H), 2.15 (br s, 2H), 1.86-1.66 (m, 2H), 1.52-1.33 (m, 2H), 1.15-0.95 (m, 1H), 0.89-0.73 (m, 3H), 0.27-0.06 (m, 1H)

[0202] LRMS [M+H] + C 16 H 20 F2N5O + : 336.24 Example 11 Alternative synthesis of compound 13 [ka]

[0203] Hydrochloric acid (12N, 22.6m, 3.35 equiv.) was added to a reaction vessel (500mL) containing water (139mL) over 90 minutes at 25°C. Isopropylamine (4.05 equiv., 25.88mL) was then added until pH 10.5. Compound 12 (25g) and pyridoxal-5'-phosphate (0.01g / g, 0.25g) were added, followed by aminotransferase: CDX-50 (0.02g / g, 0.50g). The reaction mixture was heated at 50°C and stirred for 24 hours. Hydrochloric acid (12N, 0.6 equiv., 4ml) was added, as well as 1g of Celite (4% by weight). The reaction mixture was heated at 80°C for 2 hours, then cooled to 20°C, filtered and washed. The solid residue was removed and the mixture was heated at 50°C. Potassium hydroxide solution (10 wt %, 65 mL, 1.55 equiv.) was then added until pH 9.5. The mixture was then cooled to 20° C. over 3 h. Compound 13 was isolated by filtration, rinsed with water, and dried. Example 12 Synthesis of Compound (I) [ka]

[0204] To a scintillation vial containing compound 14 (0.019 g, 0.062 mmol), HATU (33.0 mg, 0.087 mmol) in anhydrous ACN (0.5 mL) was added DBU (15 μL, 0.100 mmol). After 30 min, a solution of compound 13 (0.021 g, 0.062 mmol) in CH3CN (0.5 mL) and DMF (0.1 mL) was added. The resulting solution was stirred at room temperature for 2 h and then purified by reverse phase chromatography to give compound (I) as the trifluoroacetate salt.

[0205] 1H NMR(500MHz, CD3OD) 8.91-8.83 (m, 1H), 8.78-8.71 (m, 1H), 8.33 (s, 1H), 7.88 (d, J=2.5Hz, 1H), 7.74 (s, 2H), 7.69-7.67 (m, 1H), 7.65 (s, 1H), 7.63 (t, J=58Hz, 1H), 7.52-7.50 (m, 1H), 6.36 (d, J=0.8Hz, 1H), 6.06-5.95 (m, 1H), 2.76-2.65 (m, 1H),2.36-2.21 (m, 1H), 2.08-1.93 (m, 2H), 1.63-1.53 ​​(m, 1H), 1.53-1.42 (m, 1H), 0.99(d, J=6.9Hz, 3H)

[0206] LRMS [M+H] + C 28 H 23 Cl2F2N9O2 + : 626.09 Example 13 Preparation of compound 21

[0207] As shown in Scheme 3, compound 7 can be prepared by the following reaction steps. a) Synthesis of compound 16 [ka]

[0208] Magnesium (8.73 g, 359 mmol, 1.35 equiv.) and a crystal of I2 were added to a three-neck flask (1 L). Anhydrous THF (100 mL) was added to the reaction flask under N2. The reaction temperature was monitored with a thermometer (J-CEM). 3-Bromo-1,1-dimethoxy-propane (65.8 g, 356 mmol, 1.35 equiv.) was diluted with THF (150 mL) and added to the dropping funnel. 3-Bromo-1,1-dimethoxy-propane solution (20 mL) was added to the flask at 20 °C, and the overall light brown suspension was vigorously stirred using a stir bar and the rotation of the magnesium piece to proceed with the reaction. After 30 min, the light brown color disappeared and the solution temperature rose to 45-50 °C. The resulting reaction solution was then maintained at a temperature of 55-62 °C, and the 3-Bromo-1,1-dimethoxy-propane solution was added slowly from the dropping funnel. After 1.5 hours, the addition was complete and the entire solution was kept at 60°C for an additional 2 hours. The solution was placed in a water bath and cooled to 25°C. 4-Chloro-N-methoxy-N-methyl-pyridine-2-carboxamide; compound 15 (53.3 g, 264 mmol, 1.0 equiv) in THF (150 mL) was added to the dropping funnel. The solution substrate was added to the Grignard solution within 20 minutes to give a red-yellow solution. The internal temperature was kept below a maximum of 35°C. After 15 minutes, both HPLC and TLC showed the reaction was complete. Water (20 mL) was slowly added to the crude mixture, and a brown gel-like solid precipitated. The entire crude was filtered through Celite and washed twice with THF (total of 100 mL). The resulting crude solution was dried over Na2SO4. The resulting crude was concentrated in vacuo at 30-35°C to give compound 16 as a yellow oil.

[0209] Additional note: The crude product obtained could be used in the next reaction without purification. Spectral analysis: The crude product obtained was then purified by ISCO purification system (Hexane / EA 1:0-10:1) to give the desired product.

[0210] 1H NMR(500MHz, CDCl3)δ 8.60 (d, J=5.0Hz, 1H), 8.04 (d, J=2.0Hz, 1H), 7.49 (dd, J=5.2, 2.0Hz, 1H), 4.51 (t, J=5.6Hz, 1H), 3.35 (s, 6H), 2.10-2.05 (m, 2H), 1.66-1.60 (m, 2H)

[0211] LRMS [C 10 H 11 ClNO2] + : 212.05, 212.10 b) Synthesis of compound 18-Et

change

[0212] In a round bottom flask (250 mL), compound 16 (7.61 g, 38.5 mmol, 1 equiv) was diluted with water (20 mL) and THF (80 mL) at 20° C. To this solution, trifluoroacetic acid (8.5 mL, 110 mmol, 2.7 equiv) was added at room temperature. The solution was immediately heated at 50° C. After 4 h, the solution turned dark brown in color, and HPLC and TLC showed the reaction was complete. More water (60 mL) was added to the flask. To the resulting crude, sodium bicarbonate (9.6 g, 114 mmol, 2.8 equiv) was added slowly at 25° C. to neutralize the solvent to pH 7. The crude material was extracted three times with EtOAc (100 mL) and the combined organic crude was washed once with brine (50 mL) and dried over Na2SO4. The crude was filtered and concentrated to a dark oil, which was used directly in the next step. To the resulting crude was added CHCl (75 mL) at 20° C. To the solution containing compound 17, ethyl 2-(triphenylphosphoranylidene)propionate (14.2 g, 38.2 mmol, 0.93 equiv) was added in one portion. The reaction was kept at 20° C. for 8 h. The resulting crude was concentrated to dryness, diluted with hexanes:EtOAc (1:1), and the precipitated solid was filtered and washed twice with MTBE (20 mL). The combined crude was concentrated to a black oil and purified (hexanes / EA 1:0 to 5:1) on an ISCO purification system (200 g silica gel) to give the desired compound 18-Et (8.21 g, 70.3%) as a yellow oil.

[0213] 1 H NMR(500MHz, CDCl3)δ 8.59 (s, 1H), 8.05 (s, 1H), 7.50 (s, 1H), 6.80(s, 1H), 4.24-4.15 (m, 2H), 3.39-3.37 (m, 2H), 2.65-2.57 (m, 2H), 1.90 (s, 3H), 1.35-1.25 (m, 3H)

[0214] LRMS [C 14 H 17 ClNO3] + : 282.09, 282.21 c) Synthesis of compound 19-Et [ka]

[0215] Compound 18-Et (9.55 g, 33.9 mmol, 1.0 equiv) was added with p-toluenesulfonic acid (2.35 g, 13.5 mmol, 0.40 equiv), trimethyl orthoformate (24 mL, 220 mmol, 6.4 equiv) and methanol (95 mL) at 20° C. The solution was refluxed for 60 h. The resulting crude was cooled to 0° C., sodium hydroxide (1.7 mL, 17 mmol, 0.5 equiv) was added and the solvent was neutralized to pH 7. The crude was concentrated to a viscous oil and diluted with MTBE (200 mL). The crude was washed once with water and brine. The crude was dried over Na2SO4 and filtered. This was filtered and purified by ISCO (80 g silica) (hexane / EA 1:0 to 4:1) to give compound 19-Et (8.1 g, 73% yield) and ethyl (E)-6-(4-chloro-2-pyridyl)-2-methyl-6-oxo-hex-2-enoate (1.2 g, 13%).

[0216] 1 H NMR(500MHz, CDCl3)δ 8.50 (d, J=4.0Hz, 1H), 7.63 (d, J=0.8Hz, 1H), 7.17 (dd, J=0.8 and 4.0Hz 1H), 6.45 (dt, J =8.0 and 0.4Hz, 1H), 4.06 (q, J=7.8Hz, 2H), 3.12 (s, 6H), 2.22-2.11 (m, 2H), 1.85-1.73 (m, 2H), 1.61 (s, 3H), 1.19 (t, J=7.8Hz, 3H)

[0217] LRMS [C 16 H 22 ClNO4-OCH3] + : 297.11, 297.10 d) Synthesis of compound 20 [ka]

[0218] Pale yellow oily compound 19-Et (1.14 g, 3.48 mmol, 1.0 equiv) was diluted in ethanol (10 mL) at 20 °C. To this solution, sodium hydroxide (2 mol / L) / water (2 mL, 4 mmol, 1.1 equiv) was added at room temperature. The solution was heated at 60 °C for 12 h. HCl (1 mol / L, 4 mL) was added to the crude obtained and concentrated to give a white paste overall. Saturated aqueous NH4Cl solution (20 mL) was then added to the crude obtained and extracted twice with 2-methyl THF (10 mL). The crude obtained was dried over Na2SO4, filtered, and concentrated to give a pink crude product. The crude obtained was filtered and purified by ISCO (8 g silica) (hexane / EA 1:0 to 2:1) to give compound 20 as white crystals (1.05 g, 100%).

[0219] 1 H NMR(500MHz, CDCl3)δ 8.61 (s, 1H), 7.73 (s, 1H), 7.28 (s, 1H), 6.80-6.65 (m, 1H), 3.21(s, 6H), 2.29-2.20 (m, 2H), 1.95-1.85 (m, 2H), 1.70 (s, 3H)

[0220] LRMS [C 14 H 17 ClNO4-OCH3] + : 268.08, 268.18 e) Synthesis of compound 21 [ka]

[0221] Compound 20 (632 mg, 2.11 mmol, 1.0 equiv.) and a pressure-resistant reaction vessel were both placed in a glove box. In the glove box, the catalyst: diacetato[(R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]ruthenium(II) (95 mg, 0.109 mmol, 5.2 mol%), and methanol (5 mL) were subsequently added and the vial was placed in a reaction vessel and sealed. The reaction vessel was placed in a hydrogenation apparatus and purged with hydrogen several times. The reaction was set to 150 psi H2 at room temperature. After 12 hours, the reaction vessel was emptied and the solution had turned dark red in color. TLC and LCMS showed the reaction was complete. The crude product was concentrated and then purified by ISCO (silica, 8 g) (hexane / EA 1:0 to 1:1) to give the desired product, compound 21, as a brown solid (0.63 g, 99%). Chiral HPLC showed the desired product was 92%. %ee It was confirmed that this is the case.

[0222] 1 H NMR(500MHz, CDCl3)δ 8.59 (d, J=5.2Hz, 1H), 7.69 (d, J=1.8Hz, 1H), 7.25 (dd, J=5.2 and 1.8Hz, 1H), 3.17 (s, 6H), 2.39-2.31 (m, 1H), 2.14-2.03 (m, 2H), 1.65-1.57 (m, 1H), 1.35-1.20 (m, 1H),1.07 (d, J=7.0Hz, 3H), 0.98-0.90 (m, 1H)

[0223] LRMS [C 14 H 20 ClNO4-OCH3] + : 270.18, 270.19 Example 14 [ka]

[0224] Compound 37a (2.8 g) and methyl 2-(triphenylphosphanylidene)propanoate (11 g) were dissolved in DCM (100 mL) and stirred for 4 h at room temperature. After complete conversion, the solvent was removed under reduced pressure and the resulting residue was purified by silica gel chromatography to give the desired compound 38a (6.9 g).

[0225] 1H NMR (CDCl3, 400 MHz): δ 6.69 ppm, (1H, m); 3.72ppm (3H, s), 2.45-2.41 (2H, m), 2.36-2.32 (2H, m), 1.84 (3H, s), 1.43 (9H, s)

[0226] Compound 38a (3 g) and compound 2a (3.4 g) were dissolved in THF (30 mL) and the reaction mixture was cooled to -10°C. LiHMDS (29 mL, 2.207 eq) was then added dropwise. After complete conversion, the reaction mixture was quenched with saturated NH4Cl (150 mL) and further extracted with EtOAc (250 mL*2). The organic layer was dried over Na2SO4, filtered and concentrated. The crude compound 39a (11 g) obtained was dissolved in ACN (220 mL), water (110 mL) and methanesulfonic acid (100.5 g, 34.967 eq). The reaction was heated at 65°C until complete conversion. After cooling to room temperature, the pH was adjusted to 5-7 with concentrated NaOH solution. The organic layer was extracted with EtOAc (500 mLx2), dried over Na2SO4 and filtered to give compound 18a (10.8 g).

[0227] The obtained compound 18a was dissolved in TMSCl (24.4 g, 4.928 eq), trimethyl orthoformate (3 39 g, 8.064 eq) and MeOH (200 mL). The reaction mixture was heated at 45-50 °C and stirred overnight. After cooling to room temperature, the obtained reaction mixture was neutralized with saturated NaHCO3 solution (200 mL). The aqueous layer was extracted with DCM (3 * 400 mL), dried over Na2SO4, filtered and removed under reduced pressure. The obtained residue was purified by column silica gel chromatography (petroleum ether / EtOAc 50:1 to 3:1) to give compound 19a (10.4 g). 1H NMR (CDCl3, 400 MHz): δ 8.56ppm (1H, d, J=8Hz), 7.70ppm (1H, s), 7.24ppm (1H, dd, J=4Hz, 8Hz), 6.52ppm (1H, t, J=4Hz), 3.68ppm (3H, s), 3.18ppm (6H, s), 3.24-3.20 (2H, m), 1.86-1.83 (2H, m) Example 15 [ka]

[0228] Paraformaldehyde (1.44 g, 1.5 equiv) was charged to the reaction vessel under nitrogen followed by MeOH (25 mL, 5 vol), ethyl 2-oxocyclopentane-1-carboxylate (5 g, 1 equiv) and finally diethylamine (7.02 mL, 2.1 equiv). The reaction mixture was stirred at 20-25 °C for at least 3.5 h. After complete conversion, sodium hydroxide (6 M, 26.6 mL, 5 equiv) was added and stirred at 20-25 °C for at least 2 h until complete conversion. MeOH was distilled under vacuum. MTBE (25 mL, 5 vol) was then added to the reaction mixture and stirred for 10 min. The layers were allowed to settle and the top organic layer was removed and discarded. Aqueous HCl (6 M) was added to the bottom aqueous layer until pH 2. EtOAc (25 mL, 5 vol) was added and the reaction mixture was stirred for at least 10 min. After phase separation, it was washed with water. The organic layer was concentrated to dryness to give compound 41a. This was directly dissolved in iPrOH (50 mL, 10 vol) and H2SO4 (1 equiv.) at room temperature and stirred for 72 h. The pH of the resulting reaction mixture was adjusted to 7.3 with 20% aqueous K2HPO4. After phase separation, the iPrOH layer was diluted with 20% aqueous K2HPO4 (10 vol) and washed with toluene. Finally, the pH of the aqueous layer was adjusted to 4.7 with H2SO4 (2 M). The desired compound 42a was extracted with toluene and isolated after removal of the solvent under reduced pressure.

[0229] 1H NMR (400MHz, CDCl3): δ 6.33ppm (1H, s); 6.69 (1H, s); 5.05-4.98 (1H, m); 2.37-2.29 (4H, m), 1.87-1.80 (2H, m), 1.24 (6H, d=4Hz)

[0230] Methyl 4-chloropicolinate (0.86 g, 1 equiv) was charged to a nitrogen purged three neck flask at 20-25 °C followed by compound 42a (1 g, 1 equiv) and THF (10 mL, 10 vol) at 20-25 °C. After cooling to -30 °C, LiHMDS (1 M in THF, 12.49 mL, 2.5 equiv) was added dropwise over 30 min, keeping the temperature below -25 °C and stirring at -20 °C for at least 1 h. After complete conversion, acetic acid (0.9 mL, 3 equiv) was added dropwise, keeping the temperature below -10 °C and the resulting reaction mixture was allowed to warm to room temperature. EtOAc (50 mL, 50 vol) was added and the organic layer was washed with water and brine. After concentration under reduced pressure, a solution of water (15 mL, 15 vol) and sulfuric acid (6.7 mL, 25 equiv) was added slowly. The reaction mixture was then heated at 65° C. for at least 17 h and cooled to 20-25° C. After complete conversion, the reaction mixture was diluted with water (10 mL, 10 vol) and neutralized with 33% ammonium hydroxide to pH 4. Compound 18b was collected as a solid and dried.

[0231] 1 H NMR (500MHz, DMSO): δ 8.7ppm (1H), 7.9ppm (1H), 7.8ppm (1H), 6.0ppm (1H), 5.6ppm (1H), 3.1ppm (2H), 2.3ppm (2H), 1.8ppm (2H)

[0232] Compound 18b (5 g) was dissolved in TMOF (4 eq), H2SO4 (1.1 eq), MeOH (4 vol) and stirred overnight at 50°C. After complete conversion, NaOH (6.4 M, 8 eq) was added and stirred for 2 h. After distillation of MeOH under reduced pressure, DCM was added to the reaction mixture and the pH was adjusted to 5 with 30% aqueous citric acid. After extraction with DCM, washing with water, the organic layer was concentrated to dryness. The resulting residue was dissolved in MeCN (2.5 vol) and heated at 45°C. The reaction was slowly cooled to 0°C and stirred for 1 h. Finally, water (10 vol) was added in 2.5 vol portions. After filtration and cake washing, compound 19b was obtained as a solid in 88% yield.

[0233] 1 H NMR (500MHz, CDCl3): δ 8.6ppm (1H), 7.8ppm (1H), 7.6ppm (1H), 7.2ppm (1H), 5.5ppm (1H), 3.2ppm (6H), 2.2ppm (2H), 2.1ppm (2H), 1.1ppm (2H)

[0234] To a 5 mL vial containing a stir bar was added [RuCl(p-cymene)((R)-H8-binap)]Cl (0.0003 mmol) in a DCM stock solution (100 μL). Compound 19b (0.075 mmol) was then added as a stock solution (1 mL) in MeOH / DCM=3 / 1, followed by TEA (0.375 mmol). The vial was capped and transferred to a B48 parallel reaction vessel. The reaction was carried out under H2 atmosphere (40 bar) at 25 °C overnight (approximately 16 h). The resulting reaction was analyzed by HPLC to give the desired compound 21. Example 16 Crystallization of compound 21 [ka]

[0235] Crude compound 21 (1.0 g) was suspended in heptane (25-30 mL) and stirred at 40° C. until a clear solution was obtained. After cooling to 35° C., seed crystals from this reaction mixture were added and the mixture was stirred for 2-4 h and then cooled to −5° C. over 8-10 h. After 6-10 h at −5° C., the cake was filtered, washed, and dried in an oven at 30° C. mp: 64° C. Example 17 Enzymatic synthesis of compound 27 a) Synthesis of compounds 24a and 25a [ka]

[0236] Potassium tert-butoxide (1 mol / L) / THF (1200 g, 1330 mmol, 1 mol / L) was added to a slurry of 2-methylcyclopentanone, compound 22 (93.30 g, 931.7 mmol, 98 mass%), compound 2a (158.02 g, 902.55 mmol, 98 mass%) / THF (1500 mL, 18400 mmol, 100 mass%) at -30 ° C. The resulting yellow slurry was stirred for 1 hour between -24 and 30 ° C. In a separate reaction vessel (4 L), sulfuric acid (13.14 mol / L) / water (92 g, 660.2 mmol, 13.14 mol / L) and water (800 g, 44407.9 mmol, 100 mass%) were added and cooled to 0 ° C. in advance. The yellow slurry containing 23a-Cl was poured into the cooled acidic solution to give a slurry. The external temperature was set to 45° C. and THF was distilled at 15° C. under vacuum at 115 mbar. To the slurry (~1 liter), 500 mL of water was added. The precipitated solid was collected and the aqueous solution was removed. The collected solid was returned to the reaction vessel along with MSA (320 mL) and water (1 L). The slurry was heated at 65° C. and after 60 minutes, all solids had dissolved. The dark solution was held at 65° C. for 3 hours and cooled to room temperature to 0° C. A slurry was formed and filtered. 24a was collected and dried at room temperature to give a total of 139.8 g of beige solid. The filtrate was returned to the reaction vessel and the pH was adjusted to 5.1 with 28% by weight NH4OH. A solid formed during the pH adjustment and was filtered at room temperature. An additional 41 g of 24a was obtained as an off-white solid.

[0237] 1 H NMR (400MHz, DMSO-d6): δ 12.07 (1H, s), 8.70 (d, J=5.31Hz, 1H), 7.94 (dd, J=7.94, 1.77Hz, 1H), 7.82 (dd, J=5.18, 2.15Hz, 1H), 3.15 (m, 2H), 2.35 (m, 1H), 1.60 (m, 3H), 1.42 (m, 1H), 1.05 (d, J=6.82Hz, 3H)

[0238] LRMS [C 12 H 14 ClNO3+H] + : 258.24, 256.25

[0239] Then 178.27 g of compound 24a was added to a 2 L reaction vessel, followed by MeOH (3.4 L), TMOF (380 mL), TMSCl (210 mL). The external temperature was set to 57° C. and the mixture was heated at 49° C. After an additional 4 h at 50° C., the dark solution was cooled to 10° C. and then added to saturated NaHCO3 (2.6 L) in a 20 L reaction vessel. The total volume was 6.5 L.

[0240] Most of the MeOH solvent was distilled under vacuum at an external temperature of 35° C. and the volume was concentrated to 3.3 L. Then 2 L of MTBE was added. The organic layer was separated from the aqueous layer and concentrated to give compound 25a (207 g) as a colored liquid.

[0241] 1 H NMR (400MHz, DMSO-d6): δ 8.60 (d, J=5.05Hz, 1H), 7.60 (d, J=1.77Hz, 1H), 7.52 (dd, J=5.31, 2.02Hz, 1H), 3.50 (s, 3H), 3.02 (br s, 6H), 2.30 (m, 1H), 2.00 (m, 2H), 1.40 (m, 1H), 1.22 (m, 1H), 0.93 (d, J=7.07Hz, 3H), 0.79 (m, 2H)

[0242] LRMS [C 15 H 22 ClNO4-OCH3] + : 284.76

[0243] Kilogram Scale To a mixture of 2-methylcyclopentanone, compound 22 (235 kg, 0.66X, 1.16 equiv.) and compound 2a (354 kg, 1.0X) in 2-Me-THF (2103 L, 5.1X, 5.9 V) was added potassium tert-butoxide (258 kg, 0.73X, 1.1 equiv.) in portions over 5 h at 0 °C under N2. After 2 h, the resulting reaction mixture was quenched with water (2839 kg, 8X, 8 V; pre-cooled to 3-8 °C) over 4 h at 0 °C. The aqueous layer was separated and washed with toluene (3003 L, 2613 kg, 7.4X, 8.5 V) at 0 °C. The pH was adjusted to 7.0-9.0 (8.68) by the addition of 5% aqueous H2SO4 (1970 kg, 5.6X, 0.49 equiv) over 5.5 h at 0 ° C, and then adjusted to 4.0-6.0 (4.92) by the addition of 0.5% aqueous H2SO4 (611 kg, 1.7X, 0.02 equiv) over 2 h at 0 ° C. The mixture was stirred for 30 min at 0 ° C, then filtered by centrifugation and rinsed with water (1495 kg, 4.2X, 4.2V) to give a wet solid of compound 23 (565 kg).

[0244] Compound 23 (7.60 kg, correct assay = 7.50 g, 32.37 mmol) was added to a reaction mixture containing MSA (7.60 kg, 79.08 mol), HO (90.00 g, 90 mL) and ACN (29.25 kg, 37 L). The resulting reaction mixture was heated to 68° C. and stirred for 5 h. The reaction mixture was cooled to 20° C., then 25% ammonia solution (5.50 kg) was added in portions and stirred for 1 h. 2.5% ammonia solution (1.50 g) was added in portions to the reaction mixture over 30 min to pH 4.8. The resulting reaction mixture was heated at 43° C. for 8 h and filtered to give compound 24a (40 kg).

[0245] Compound 24a (37.0 kg) was mixed with 3.0 equivalents of CH(OMe)3 and 2.0 equivalents of TMSCl in MeOH (370 L). After stirring the mixture for 24 h at 30-35 °C, the resulting reaction mixture was cooled to 20-25 °C and quenched with 2.2 equivalents of TEA at 20-30 °C. The reaction mixture was then concentrated to 100 L under vacuum at below 40 °C. MTBE (370 L) and H2O (300 L) were added to the residue. After phase separation, the organic layer was collected and H2O (200 L) was used to wash the organic layer. The organic layer was concentrated to 70 L under vacuum at below 40 °C. DMSO (1 V) was then added to the residue and the mixture was concentrated to 70 L under vacuum at below 40 °C to obtain a concentrated DMSO solution of compound 25a (86.4 kg). b) Enzymatic resolution of dimethoxymethyl esters: [ka]

[0246] A reaction vessel (250 mL) was used with a water circulator to maintain the reaction temperature at 35° C. throughout the reaction sequence. The pH was maintained at 7.0 using a calibrated pH meter. To a 250 mL reaction vessel was added compound 25a (5.0 g), DMSO (4.0 mL, 2%), and sodium phosphate buffer (180 mL, 0.1 M, pH 7.0). The reaction solution was kept at pH 7.0 using continuous 5N sodium hydroxide. Lipase MH Amano 10 SD (1.0 g) was dissolved in the above buffer (10 mL) and added to the reaction vessel. An additional 6.0 mL of buffer was used to rinse the enzyme vessel and added to the same reaction vessel. Samples (80 μL) were pipetted, diluted with methanol (1.920 mL), vortexed, centrifuged for 2 min, filtered, and analyzed by achiral and chiral HPLC. The reaction was stopped after 23 hours and the pH was adjusted to 8.2 with 10N sodium hydroxide. The resulting reaction solution was extracted with ethyl acetate (200 mL). All dimethoxymethyl esters were extracted into the organic layer. The organic layer was back-extracted with pH 8.5 sodium potassium buffer (2x50 mL) to remove acid. The organic layer was washed with brine (50 mL), water (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered, the solvent removed under reduced pressure and the resulting residue was dried in a vacuum oven overnight. Compound 26a (1.88 g) was isolated as a brownish liquid (yield 37.6%, ee 98.7%).

[0247] 1 H NMR(400MHz, DMSO-d6)δ 8.61 (d, J=5.1Hz, 1H), 7.60 (d, J=1.5Hz, 1H), 7.52 (dd, J=5.2, 2.1Hz, 1H), 3.50 (s, 3H), 3.03 (s, 3H), 3.03 (s, 3H), 2.31 (sxt, J=7.0Hz, 1H), 2.05-1.95 (m, 2H), 1.47-1.35 (m, 1H), 1.28-1.17 (m, 1H), 0.94 (d, J=7.1Hz, 3H), 0.85-0.74 (m, 2H)

[0248] LRMS [C15 H 22 ClNO4-OCH3] + : 284.2 / 286.1 c) Hydrolysis of compound 26a to give compound 27a:

[0249] To a reaction vial were added compound 26a (1.0 g), methanol (20 mL), water (5.0 mL), and 10N sodium hydroxide (500 μL, 0.2 g). The reaction was stirred at room temperature for 5 h. A sample (30 μL) was removed, diluted with methanol (970 μL), vortexed, filtered, and analyzed by HPLC. Most of the reaction (~98% conversion) was complete within 2 h, and no racemization was observed during hydrolysis. The reaction mixture was concentrated to an oil and diluted with water (20 mL, pH 12.7). The resulting reaction mixture was extracted with MTBE (2x50 mL) to remove MTBE. The aqueous layer was cooled and acidified to pH 3.8 with 6N HCl (850 μL). The aqueous layer was extracted with MTBE (2x50 mL), and the MTBE solution was washed with brine (25 mL) and water (2x25 mL). The MTBE solution was dried over anhydrous sodium sulfate, filtered, the solvent removed, and the resulting residue was dried in a vacuum oven overnight. Compound 27a was isolated as a viscous yellow liquid (940 mg, 98.3% yield, AP 97 and ee 98%).

[0250] 1 H NMR(400MHz, DMSO-d6)δ 11.96 (s, 1H), 8.60 (d, J=5.3Hz, 1H), 7.60 (d, J=2.0Hz, 1H), 7.51 (dd, J=5.2, 2.1Hz, 1H), 3.04 (s, 3H), 3.01 (s, 3H), 2.18 (sxt, J=6.9Hz, 1H), 2.00 (dd, J=10.6, 5.8Hz, 2H), 1.47-1.33 (m, 1H), 1.26-1.13 (m, 1H), 0.91 (d, J=6.8Hz, 3H), 0.87-0.75 (m, 2H)

[0251] LRMS [C 14 H20 ClNO4] - : 300.1 Alternative to enzymatic resolution [ka]

[0252] 0.1 M Tris buffer (13.5 L) supplemented with Ca(OAc)2 (50 mM) was mixed with 2.5 wt% enzyme (Almac Hydrolase L90 enzyme, also known as AL-L90, available from ALMAC Group Ltd., Craigavon, Northern Ireland, UK) at 20-25 °C. The pH of the mixture was adjusted to 7.2-7.8. The mixture was heated at 38-42 °C and kept at pH 7.2-7.8. A solution of compound 25a (1.5 kg) in DMSO (1.5 L) was added in one portion to the above mixture at 38-42 °C. After 22 h, the conversion was 49%. The reaction mixture was cooled to 5 °C and kept at 0-10 °C for 16 h. ACN (9 L) was added to the reaction mixture at 0-10 °C for workup. The pH was then adjusted to 10.0 with 20% K2CO3 and 0.25X Celite was added to the mixture. After stirring for 20-30 minutes, the resulting reaction mixture was filtered and the cake was rinsed with MTBE (3 L) and water (3 L). The filtrate was collected and MTBE (15 L) was added to the mixture and the phases were separated. The organic layer was washed twice with water (15 L). The aqueous layers were combined and adjusted to pH 5.5 with 20% citric acid. MTBE (15 L) was added to the mixture and the phases were separated. The aqueous layer was then adjusted to pH 5.5 with 20% citric acid and phase separated again with MTBE (15 L). The organic layers were combined and washed with process water (15 L). The organic layers were collected, filtered and the aqueous layer was removed. The organic layer was then concentrated to 4.5 L. Azeotroped twice with 15 L MTBE to 4.5 L. 7.5 L of MTBE and 3 L of ACN were added to the resulting residue to form the salt. In the salt formation, the mixture was heated at 50-55 °C and DCHA (0.75 equiv.) was added to the mixture at 50-55 °C. After stirring at 50-55 °C for 1 h, the resulting reaction mixture was cooled at 10 °C / h. 1.8% seed crystals were added to the mixture at 38.2 °C. A white solid precipitated slowly. After holding at 38 °C for 3 h, the mixture was continued to be cooled at 10 °C / h. After holding at 0-5 °C for 10 h, the mixture was filtered. Pre-chilled ACN (1.5 L) was used to rinse the cake. After drying for 38 h, 1.245 kg of compound 27a-DCHA was obtained as a white solid (99.3% purity, 99.4 ee% and 43.61% isolated yield). Example 18 Synthesis of compound 10 [ka] a) Synthesis of compound 29a

[0253] To a solution of (triisopropylsilyl)acetylene (10.5 g, 57.6 mmol, 100 wt%) in THF (60 mL) was slowly added n-butyllithium (2.5 mol / L) in hexane (22 mL, 55.0 mmol, 2.50 mol / L) at -10°C. After the addition, the mixture was heated at 21°C. A solution of compound 28a (10.0 g, 49.8 mmol, 100 wt%) in THF (35 mL) was then added at 21°C. After 1 h, HPLC analysis indicated that 8% of the amide starting material remained. Lithium bis(trimethylsilyl)amide (1 M, in THF, 8 mL) was then added. After 1 h, HPLC analysis indicated the reaction was complete. The mixture was cooled to 0-5°C and added to a mixture of 15% aqueous citric acid (500 g) and heptane (0.6 L) at 5-15°C. The organic layer was washed with 3% aqueous citric acid (200 mL) and water (0.2 L), dried over MgSO4, and concentrated to give compound 29a (15.8 g) as an orange oil (98.5% yield).

[0254] 1 H NMR (500MHz, CDCl3): δ 8.68 (1H, d, J=4.5Hz), 8.15 (1H, s), 7.50 (1H, d, J=4.5Hz), 1.25-1.05 (21 H, m)

[0255] LRMS [C 17 H 24 ClNOSi+H] + : 322.23 / 324.11 b) Synthesis of compound 30a

[0256] To a solution of compound 29a (3.00 g, 9.32 mmol) in MeOH (20 mL) was added trimethyl orthoformate (2.0 mL, 18 mmol) followed by chlorotrimethylsilane (3.0 mL, 24 mmol) at 21 °C. The mixture was then heated at 60 °C. After 1 h, HPLC analysis indicated that the starting material was consumed. The mixture was cooled to 21 °C and added to a mixture of hexane (200 mL) and a solution of NaHCO3 (15 g) and Na2CO3 (5 g) in water (200 mL). The separated organic layer was then dried over MgSO4 and concentrated to give [3-(4-chloro-2-pyridyl)-3,3-dimethoxy-prop-1-ynyl]-triisopropyl-silane (3.45 g, 9.38 mmol, 100% yield) as an orange oil. The resulting crude intermediate was used for subsequent deprotection of TIPS without further purification.

[0257] To a solution of [3-(4-chloro-2-pyridyl)-3,3-dimethoxy-prop-1-ynyl]-triisopropyl-silane (3.20 g, 8.70 mmol, 100 wt%) in 2-MeTHF (10 mL) and TBME (10 mL) was added 1 M tetrabutylammonium fluoride (THF solution, 12 mL, 12.0 mmol, 1.00 mol / L) at 21° C. After 10 min, HPLC analysis showed that the starting material was consumed. The dark mixture was added to a mixture of TBME (0.2 L) and an aqueous solution of K2HPO4 / K3PO4 (20 g / 5 g / 130 mL water). The isolated organic layer was dried over MgSO4 and concentrated. The resulting residue was purified by column chromatography (20-60% EtOAc / heptane; Rf 0.39 in 50% EtOAc / heptane) to give compound 30a (1.65 g) as a pale solid in 90% yield.

[0258] 1 H NMR (500MHz, CDCl3): δ 8.61 (1H, d, J=5.0Hz), 7.75 (1H, s), 7.32 (1H, d, J=5.0Hz), 3.37 (6H, s), 2.74 (1H, s)

[0259] LRMS [C 10H 10 ClNO2-OCH3] + :180.25 / 182.06 c) Synthesis of compound 31a

[0260] To a solution of bis(cyclopentadienyl)zirconium dichloride (20.7 g, 70.9 mmol, 99 wt%) in THF (200 mL), lithium tri-tert-butoxyaluminum hydride (1 mol / L) in THF (71 mL, 71 mmol, 1 mol / L) was added at 7-15°C and stirred at 0-5°C for 1 hour. Compound 30a (12.5 g, 59.1 mmol, 100 wt%) was then added at 0-5°C. After mixing at 5-10°C for 10 minutes, the mixture was heated to 21°C and stirred at the same temperature for 0.5 hours. The dark brown solution mixture was then cooled to 0-5°C, and NBS (11 g, 61.8 mmol, 100 wt%) was added as a solid in two portions (6 g first, 5 g second).

[0261] After stirring at 5-10°C for 2 h, the reaction mixture was added to a mixture of EtOAc / heptane (180 mL / 60 mL) and 15% aqueous NH4Cl (250 mL). The resulting slurry was filtered through Celite and the filter line was rinsed with EtOAc (30 mLx2). The aqueous layer was removed from the combined filtrate. The resulting organic layer was washed with 5% aqueous K2HPO4, dried over MgSO4, and concentrated. The residue was purified by column chromatography (0-30% EtOAc / heptane; product Rf 0.4 in 30% EtOAc / heptane) to give compound 31a (13.2 g, 45.1 mmol, 76.4% yield) as a grey solid.

[0262] 1 H NMR (500MHz, CDCl3): δ 8.57 (1H, d, J=5.2Hz), 7.67 (1H, s), 7.26 (1H, d, J=5.2Hz), 6.78 (1H, d, J=13.5Hz), 6.11 (1H, d, J=13.5Hz), 3.24 (6H, s)

[0263] LRMS [C 10 H 11BrClNO2-OCH3-Br] + : 182.09 / 184.22 d) Synthesis of compound 32a

[0264] To a solution of compound 31a (12.0 g, 41.0 mmol, 100 wt%) in THF (50 mL) was added (S)-(-)-3-methoxy-2-methyl-3-oxopropylzinc bromide (0.5 mol / L) in THF (94 mL, 47 mmol, 0.50 M) at 5-10 °C. The mixture was degassed by bubbling with N2 for 3 min, and then 1,1'-bis(di-tert-butylphosphino)ferrocenedichloropalladium (0.68 g, 1.03 mmol, 100 wt%) was added at 5-10 °C. The resulting mixture was degassed by bubbling with N2 for 5 min. After 15 min at 5 °C, the dark solution was warmed to 21 °C. After 16 h, TBME (150 mL) was added to the reaction mixture, followed by aqueous NH4Cl (25%, 200 g). The dark organic layer was dried over MgSO4 and concentrated, and the resulting residue was purified by column chromatography (5-60% EtOAc / heptane; Rf 0.28 in 1:1 EtOAc / heptane) to give compound 32a (11.7 g, 37.3 mmol, 100% by weight, 90.9% yield) as an orange oil.

[0265] 1 H NMR (500MHz, CDCl3): δ 8.54 (1H, d, J=5.2Hz), 7.65 (1H, s), 7.22 (1H, d, J=5.2Hz), 5.98 (1H, dt, J=15.6, 7.3Hz), 5.45 (1H, d, J=15.6Hz), 3.62 (3H, s), 3.21 (6H, s), 2.63-2.52 (1H, m), 2.32-2.45 (1H, m), 2.16-2.20 (1H, m), 1.13 (3H, d, J=7.0Hz)

[0266] LRMS [C 15 H 20 ClNO4-OCH3] + : 282.19 / 284.23 e) Synthesis of compound 33a

[0267] In the preparation of the catalyst, diacetoxypalladium (440 mg, 1.960 mmol) and bis(1-adamantyl)-butyl-phosphane (708 mg, 1.975 mmol) were added to dioxane (12 mL). The mixture was degassed by bubbling with N2 for 0.5 h. Compound 32a (6.2 g, 20 mmol, 100 wt%), compound 8 (4.0 g, 25 mmol), pivalic acid (1.1 g, 11 mmol, 100 wt%), potassium carbonate (8.1 g, 59 mmol) were mixed in dioxane (60 mL) and the resulting mixture was degassed by bubbling with N2 for 0.5 h. The above premixed and degassed catalyst was then transferred to the mixture containing the substrate. The resulting mixture was degassed by bubbling with N2 for 0.5 h, then heated at 90 °C and held at the same temperature for 3 h, followed by cooling to 21 °C. The resulting mixture was filtered and the filter line was rinsed with TBME (25 mL). The combined filtrate was concentrated. The resulting residue was purified by column chromatography (Rf 0.36 in 60% EtOAc / heptane; 10-80% EtOAc / heptane) to give compound 33a (7.6 g, 16 mmol, 98% by weight, 86% yield) as a brown oil.

[0268] 1 H NMR (500MHz, CDCl3): δ 8.86 (1H, d, J=4.7Hz), 8.33 (1H, s), 7.73 (1H, s), 7.22 (1H, d, J=4.7Hz), 7.13 (1H, t, J=57.6Hz), 5.97 (1H, dt, J=15.7, 7.6Hz), 5.53 (1H, d, J=15.7Hz), 3.59 (3H, s), 3.23 (6H, s), 2.65-2.52 (1H, m), 2.35-2.48 (1H, m), 2.19-2.25 (1H, m), 1.12 (3H, d, J=7.0Hz)

[0269] LRMS [C 19 H 22 F2N4O6-OCH3] + : 409.18 f) Synthesis of compound 10a

[0270] A 100 mL pressure flask was charged with a solution of compound 33 (6.3 g, 14 mmol) in MeOH (50 mL) and a stir bar. The vessel was evacuated and backfilled with N2 six times. 1.35 g (0.634 mmol, 5% by weight) of Pd / carbon (10% by weight, 50% wet) was then added. The vessel was degassed with N2 and then H2. Hydrogen gas pressure was set at 80 psi and reaction temperature at 55° C. After maintaining the pressure and temperature for 12 hours, the mixture was filtered and the filtration line was rinsed with MeOH (35 mL). The combined filtrate was concentrated. The resulting residue was dissolved in THF (60 mL) and aqueous NaOH (1 mol / L) was added. The mixture was heated at 40° C. and maintained at the same temperature for 6 hours. The mixture was then cooled to 21° C. 2-MeTHF (100 mL) was added. The pH of the mixture was adjusted to pH~6 by adding 85% H3PO4. The aqueous layer was removed and the organic layer was dried over MgSO4 and concentrated. The resulting residue was purified by column chromatography (0~10% MeOH / DCM; Rf 0.4 in 10% MeOH / DCM) to give compound 10a (4.85 g, 12.2 mmol, 89.7% yield) as a foam solid.

[0271] 1 H NMR (500MHz, CDCl3): δ 8.77 (1H, s, br), 7.75 (1H, s. br), 7.39 (1H, s. br), 7.33 (1H, s. br), 7.11 (1H, t, J=59.1Hz), 5.75 (3H, s, br), 3.17 (6H, s), 2.35-2.25 (1H, m), 2.20-1.98 (2H, m), 1.55-1.43 (1H, m), 1.30-1.13 (1H, m), 0.80-1.09 (5H, m)

[0272] LRMS [C 18 H 24 F2N4O4+H] + : 399.18 Example 19 Synthesis of compound 14 [ka] Suzuki Coupling

[0273] Acetonitrile (500 mL, 5 L / kg), compound 43 (100 g, 1.0 equiv.), compound 44 (60 g, 1.05 equiv.), 1,1,3,3-tetramethylguanidine (93.2 g, 2.05 equiv.) and water (70.8 g, 10 equiv.) were added to a 2 L reaction vessel. The headspace of the reaction vessel was purged using a stream of nitrogen. Pd(dppf)Cl2·DCM (3.2 g, 1 mol%) was added to the resulting reaction mixture and the reaction vessel was heated at 65 °C for 1-7 h and stirred at that temperature for 1-17 h. After the reaction was complete, 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione sodium salt (TriNaTMT, 10 g, 10 w / w%) was dissolved in water (50 mL, 0.5 L / kg) and added to the resulting reaction mixture. To this reaction mixture was then slowly added warm water (280 mL, 2.8 L / kg) over 0.5-4 hours while maintaining the internal temperature at 60-65°C. After standing for 1-4 hours, the reaction mixture was then slowly added warm water (350 mL, 3.5 L / kg) over 2-6 hours while maintaining the internal temperature at 60-65°C. The resulting reaction mixture was then cooled to 10-15°C over 4-6 hours and further stood at 10-15°C for 1-3 hours. The slurry was then filtered and the cake was washed with cold aqueous MeCN (2:1 v / v%, 5 L / kg). The cake of compound 45 was dried in vacuum at 45-50°C overnight.

[0274] Typical results: Yield 90-95%, purity 98a%, assay >95% PD removal

[0275] In a 5 L reaction vessel, crude compound 45 solid (100 g, 1.0 equiv.), 2-methyltetrahydrofuran (MeTHF, 1.5 L, 15 L / kg) and N-acetyl L-cysteine ​​aqueous solution (32 g / 1.5 L of water) were added. The resulting mixture was stirred at 20-30 °C for 1-2 h and filtered through Celite (30 g, 0.3 kg / kg). The biphasic mixture was left for 0.5-2 h and the two layers were separated. The upper layer was left in the reaction vessel and further N-acetyl L-cysteine ​​aqueous solution (32 g / 1.5 L of water) was added. The resulting mixture was stirred at 20-30 °C for 1-2 h and the biphasic mixture was left for 0.5-2 h and the two layers were separated. The upper layer was left in the reaction vessel and further sodium bicarbonate aqueous solution (70 g / 1 L of water) was added. The resulting mixture was stirred at 20-30°C for 1-2 hours, the biphasic mixture was allowed to stand for 0.5-2 hours, and the two layers were separated. The upper layer was left in the reaction vessel, and an aqueous solution of sodium sulfate (100 g / 1 L of water) was added. The resulting mixture was stirred at 20-30°C for 1-2 hours, the biphasic mixture was allowed to stand for 0.5-2 hours, and the two layers were separated. The upper layer was left in the reaction vessel, and the solution was diluted with MeTHF (1 L, 10 L / kg), and the resulting solution was concentrated in vacuum to 500-600 mL (5-6 L / kg). The solution was then diluted with MeTHF (1 L, 10 L / kg), and the resulting solution was concentrated in vacuum to 500-600 mL (5-6 L / kg). The solution was then diluted with MeTHF (1 L, 10 L / kg), and the water content was measured (KF: 0.1% or less).

[0276] Typical results: 90-95% yield, 98a% purity Azidation / Click reaction

[0277] A solution of compound 45 in MeTHF (100 g / 1.5 L of MeTHF), MeTHF (1.5 L, 15 L / kg) and MeCN (1 L, 10 L / kg) were added to the reaction vessel and the mixture was cooled to 5-10 °C. TMSN3 (59.0 g, 1.2 equiv.) was added slowly to the reaction vessel. tBuONO (53.0 g, 1.2 equiv.) was added slowly to the reaction vessel and the mixture was stirred at 5-15 °C for 4-8 h. Aqueous NaOH (100 g / 1 L of water) was added slowly to the above mixture. This was then warmed to 20-30 °C and stirred for 20-30 min and left at 20-30 °C for 30-60 min. After phase separation, the upper layer was left in the reaction vessel and aqueous NaOH (100 g / 1 L of water) was added slowly to the above mixture. This was then stirred for 20-30 minutes and left at 20-30°C for 30-60 minutes. After phase separation, the top layer was left in the reaction vessel and an aqueous NaOH solution (100g / 1L water) was slowly added to the mixture. This was then stirred for 20-30 minutes and left at 20-30°C for 30-60 minutes. After phase separation, the top layer was left in the reaction vessel and an aqueous NaOH solution (100g / 1L water) was slowly added to the mixture. This was then stirred for 20-30 minutes and left at 20-30°C for 30-60 minutes. After phase separation, the top layer was left in the reaction vessel and the remaining azide was measured (residual N3<3ppm). An aqueous sodium sulfate solution (100g / 1L water) was then added to the mixture. This was then stirred for 20-30 minutes and left at 20-30°C for 30-60 minutes. After phase separation, aqueous sodium sulfate solution (100g / 1L water) was added to the above mixture. This was then stirred for 20-30 minutes and allowed to stand at 20-30°C for 30-60 minutes. After phase separation, aqueous sodium sulfate solution (100g / 1L water) was added to the above mixture. This was then stirred for 20-30 minutes and allowed to stand at 20-30°C for 30-60 minutes. After phase separation, aqueous sodium sulfate solution (100g / 1L water) was added to the above mixture. This was then stirred for 20-30 minutes and allowed to stand at 20-30°C for 30-60 minutes. After phase separation, the pH was measured (pH<9) and the organic azide solution was cooled to 5-15°C. After bubbling with nitrogen for 20-40 minutes, the temperature was kept at 5-15°C while triethylamine (95.0g, 2.2 equiv.) was slowly added to the reaction mixture.Trimethylsilylacetylene (50.0 g, 1.2 equiv.) was slowly added to the reaction mixture while maintaining the temperature at 5-15°C and purging the reaction vessel with nitrogen until the oxygen level was below 0.1%. Copper iodide (8.0 g, 10 mol%) was charged to the reaction vessel, which was then purged with nitrogen again until the oxygen level was below 0.1%. The resulting reaction mixture was stirred at 5-15°C for 8-16 hours. After completion of the reaction, the reaction mixture was warmed to 20-30°C and 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione sodium salt (TriNaTMT, 10 g, 10 w / w%) was added. After stirring at 20-30°C for 0.5-1.5 hours, the mixture was filtered through Celite (30 g, 0.3 kg / kg) and the cake was rinsed with MeTHF (250 mL, 2.5 L / kg). Aqueous ammonia solution (100 g / 1 L of water) was added to the reaction vessel, which was then stirred at 20-30 °C for 20-30 min and allowed to stand at 20-30 °C for 30-60 min. After phase separation, aqueous sodium sulfate solution (100 g / 1 L of water) was added to the reaction vessel, which was then stirred at 20-30 °C for 20-30 min and allowed to stand at 20-30 °C for 30-60 min. After phase separation, the upper layer was filtered through Celite (30 g, 0.3 kg / kg). After washing the Celite cake with MeTHF (0.5 L, 5 L / kg), the mixture was concentrated under reduced pressure at 45 °C to 500-700 mL (5-7 L / kg). n-Heptane (1 L, 10 L / kg) was added dropwise to the reaction vessel, and the resulting mixture was concentrated under reduced pressure at 45 °C to 500-700 mL (5-7 L / kg). n-Heptane (1 L, 10 L / kg) was added dropwise to the reaction vessel and the resulting mixture was concentrated under reduced pressure to 500-700 mL (5-7 L / kg) at 45 °C. The reaction mixture was then warmed to 55-60 °C and stirred at the same temperature for 2-4 h. After cooling the reaction vessel to 5-15 °C over 3-8 h, the slurry was aged at 5-15 °C and filtered. The cake of compound 46 was washed with n-heptane (1 L, 10 L / kg) and dried under reduced pressure at 40-45 °C for 6-12 h.

[0278] Typical results: Yield 85-90%, purity 98a%, assay >95% Chlorination

[0279] Compound 46 (100 g, 1.0 equiv.), DMF (500 mL, 5 L / kg) were added to the reaction vessel, cooled to -15°C to -5°C, and purified with water (5 g, 1.0 equiv.). 1,3-dichloro-5,5-dimethylhydantoin (DCDMH, 13.7 g, 0.75 equiv.) was added in portions while the internal temperature was kept below 5°C. The internal temperature was then adjusted to 0-10°C, and the mixture was stirred at the same temperature for 5-12 hours. After the reaction was complete, water (70 mL, 0.7 L / kg) was added to the resulting reaction mixture over 0.5-1.5 hours. During this time, the internal temperature was kept below 15°C. Seed crystals of compound 47 (0.1 g, 0.001 kg / kg) were added to the resulting reaction mixture, which was then aged at 0-10°C for 1-2 hours. Water (530 mL, L / kg) was added over 3-8 h at 0-10 °C and the slurry was aged at 0-10 °C for 4-6 h. The mixture was filtered and the cake was washed with cold water (0.5 L, 0-10 °C). The resulting crude compound 47 was dried under reduced pressure at 40-50 °C for 8-15 h.

[0280] Typical results: Yield 90-95%, purity 98a%, assay >95% Recrystallization of Compound 47

[0281] Crude compound 47 (100 g, 1.0 equiv.), DCM (0.5 L, 5 L / kg) were added to the reaction vessel and the resulting solution was stirred at 20-30 °C for 0.5-2 h. The resulting mixture was filtered through a charcoal-packed filter cartridge, circulated for 3-8 h, and then concentrated under reduced pressure to 300-360 mL (3.0-3.6 L / kg). The mixture was then warmed to 35-45 °C and refluxed for 20-40 min, before being cooled to 0-10 °C over 1-4 h and further aged at 0-10 °C for 0.5-2 h. n-Heptane (1.6 L, 16 L / kg) was then charged to the reaction vessel over 1-3 h at 0-10 °C and the slurry was aged at 0-10 °C for 1-3 h. After filtering the slurry, the cake of pure compound 47 was washed with n-heptane (500 mL, 5 L / kg) and dried under reduced pressure at 40-50 °C for 6-12 h.

[0282] Typical results: Yield 90-95%, Purity >99.5a%, Assay >95% Demethylation

[0283] Pure compound 47 (100 g, 1.0 equiv.), aqueous HCl (35 w / w%, 320 g, 10 equiv.) were added to the reaction vessel, the reaction vessel was warmed to 40-50 °C for 1-3 h, and stirred at the same temperature for 10-18 h. After completion of the reaction, aqueous ammonia (50 g / 500 mL) was added dropwise to the reaction vessel at 40-50 °C for 2-6 h until pH=5-7. The reaction mixture was then cooled to 0-10 °C for 1-3 h, and aged at the same temperature for 1-3 h. After filtering the slurry, the resulting cake of crude compound 14 was washed with cold water (1 L, 0-10 °C, 10 L / kg) and dried under reduced pressure at 40-50 °C for 12-24 h.

[0284] Typical results: Yield 90-95%, Purity >99.5a%, Assay >95% Recrystallization of compound 14

[0285] Crude compound 14 (100 g, 1.0 equiv.) and acetone (1.4 L, 14 L / kg) were added to a reaction vessel, the reaction vessel was warmed to 50-60 °C, and stirred at the same temperature for 1-3 h. Crystals of compound 14 (0.5 g, 0.005 kg / kg) were seeded, and n-heptane (1.7 L, 17 L / kg) was added at 50-60 °C over 4-8 h, the mixture was stirred at the same temperature for 1-2 h, and cooled to 5-15 °C over 2-4 h. After filtration, the cake of pure compound 14 was washed with cold n-heptane (0.5 L, 5 L / kg) and dried under vacuum at 80-90 °C for 8-16 h.

[0286] Typical results: Yield 90-95%, Purity >99.9a%, Assay >95%

Claims

1. Compound (I): 【Chemistry 1】 A method for producing a compound comprising the steps of: 1) Compound 1: 【Chemistry 2】 Compound 2: 【Chemistry 3】 [In the formula, X is selected from Cl, Br, and I; Y is OR 9 , N.H.O.C. 1-3 alkyl, Cl, Br, and I; and R 9 is C 1-3 Alkyl, C 1-3 hydroxyalkyl, substituted phenyl and substituted benzyl in a suitable solvent to give compound 3a or 3b: 【Chemistry 4】 get; 2) Compound 3a or 3b is converted in the presence of an acid to give compound 4: 【Chemistry 5】 get; 3) Compound 4 is then reacted with trialkyl orthoformate in an alcoholic solvent to give compound 5: 【Chemistry 6】 [In the formula, R 1 is C 1-6 is alkyl; and R 3 ' is C 1-6 alkyl, optionally substituted phenyl and benzyl; 4) The ester of compound 5 is hydrolyzed under basic conditions, or R 3 When ' is a substituted benzyl, hydrogenolysis is carried out to give compound 6: 【Chemistry 7】 (In the formula, R 1 and X is defined as above); 5) The carboxyl moiety of compound 6 is then activated and reacted with the chiral auxiliary HXa to give compound 6a: 【Chemistry 8】 [In the formula, Xa is 【Chemistry 9】 is selected from, where R 6 is selected from C 1-3 alkyl, phenyl, benzyl; R 7 is selected from H and phenyl; and R 8 is selected from C 1-3 alkyl, phenyl, and benzyl; R 1 and X is defined as above; 6) Compound 6a is then reacted with a base in the presence of a methyl donor selected from alkyl halides to produce compound 6b: 【Chemistry 10】 (In the formula, R 1 , X, and Xa are defined as above); 7) Removal of Xa gives compound 7: 【Chemistry 11】 (In the formula, R 1 and X is defined as above); 8) Compound 7 is then reacted with compound 8 in the presence of a metal catalyst: 【Chemistry 12】 to give compound 9: 【Chemistry 13】 get; 9) The nitro group of compound 9 is reduced to give compound 10: 【Chemistry 14】 get; 10) Compound 10 is cyclized with a suitable coupling agent to give compound 11: 【Chemistry 15】 get; 11) The ketone group is deprotected in the presence of acid to give compound 12: 【Chemistry 16】 get; 12a) reducing compound 12 with an equivalent amount of ammonia in the presence of a reducing agent; or 12b) Compound 12 is reduced by aminotransferase in the presence of an amine donor selected from isopropylamine, alanine, 3-aminobutyric acid and methylbenzylamine, a different recyclization system selected from aminotransferase / lactate dehydrogenase / glucose dehydrogenase and aminotransferase / amino acid dehydrogenase / formate dehydrogenase, and a cofactor to give compound 13: 【Chemistry 17】 to obtain an amine stereocenter represented by 13) Compound 13 is then reacted with compound 14: 【Chemistry 18】 to form compound (I): 【Chemistry 19】 get; A method comprising the steps of:

2. The method of claim 1, wherein in step 2, the acid is selected from sulfuric acid, methylsulfonic acid (MSA), benzenesulfonic acid, nitric acid, hydrochloric acid, trichloroacetic acid, and perchloric acid.

3. The chiral auxiliary HXa is 【Chemistry 20】 The method of claim 1, wherein the compound is selected from the group consisting of 4. The method of claim 1, wherein in step 8, the metal catalyst comprises palladium and a phosphine ligand.

5. 5. The method of claim 4, wherein the phosphine ligand is XPhos or SPhos.

6. The method of claim 1, wherein in step 10, the coupling agent is chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH).

7. The method of claim 1, wherein in step 12b, the aminotransferase is selected from ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260.

8. The method of claim 1, wherein in step 12b, the transamination is carried out in the presence of (4-formyl-5-hydroxy-6-methylpyridin-3-yl)methyl phosphate (PLP).

9. The method of claim 1, wherein in step 12b, the amine donor is isopropylamine.

10. Additionally, compound 14: 【Chemistry 21】 2. The method of claim 1 comprising producing 1) Compound 43: 【Chemical 22】 Compound 44: 【Chemistry 23】 in a suitable solvent under suitable Suzuki coupling conditions to give compound 45: 【Chemistry 24】 get; 2) Compound 45 is converted to an azide, which is reacted with trimethylsilylacetylene and a metal catalyst in a suitable solvent to give compound 46: 【Chemistry 25】 get; 3) Compound 46 is reacted with 1,3-dichloro-5,5-dimethylhydantoin in a suitable solvent to give compound 47: 【Chemistry 26】 obtain; and 4) Compound 47 is demethylated in hydrochloric acid to give compound 14; The method of claim 1, characterized by the steps:

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