Novel synthesis options for preparing compounds

Abstract

The present invention provides efficient processes for the preparation of key intermediates in the synthesis of Compound (I), which can be widely used and can provide selected components with various substituents. # imgabs0 #

Description

This application is a divisional application of an invention patent application with an application date of April 10, 2020, application number 202080041935.8 (international application number PCT / US2020 / 027655), and name “Novel synthetic options for preparing compounds”. Technical Field

[0001] The present invention generally relates to certain improved methods for preparing (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-(methylene-bridged)pyrazolo[4,3-b][1,7]diazacyclotetradecyn-5(6H)-one, which are FXIa inhibitors useful in treating thrombotic disorders, including venous thrombosis and deep vein thrombosis. Background Art

[0002] Factor XIa is a plasma serine protease involved in regulating blood coagulation, which is generated in vivo by combining tissue factor (TF) with factor VII (FVII) to generate factor VIIa (FVIIa). The resulting TF:FVIIa complex activates factor IX (FIX) and factor X (FX), which lead to the generation of factor Xa (FXa). The generated FXa catalyzes the conversion of prothrombin to a small amount of thrombin, which is then closed by the tissue factor pathway inhibitor (TFPI). The coagulation process is then further developed by activating factors V, VIII and XI through feedback of catalytic amounts of thrombin. (Gailani, D. et al., Arterioscler. Thromb. Vasc. Biol., 27: 2507-2513 (2007)). The resulting release of thrombin converts fibrinogen to fibrin, which polymerizes to form the structural framework of the blood clot and activates platelets, which are key cellular components of coagulation (Hoffman, M., Blood Reviews, 17: S1-S5 (2003)). Therefore, factor XIa plays a key role in developing this amplification loop and is therefore an attractive target for antithrombotic therapy.

[0003] U.S. Patent No. 9,453,018 discloses macrocyclic compounds as factor XIa inhibitors useful for treating thrombotic disorders. One of the compounds has the following structure:

[0004] The US patent discloses a multi-step synthetic method for preparing macrocyclic compounds. The method includes coupling a pyridine-containing macrocycle with a pyrimidinol to form compound (I). The disclosed method also includes a closed-loop metathesis method using a catalyst such as Grubbs (II).

[0005] The multi-step synthesis disclosed in U.S. Pat. No. 9,453,018 is difficult to adapt to large-scale synthesis, such as pilot plant or manufacturing scale production. One difficulty is that the Grubbs (II) reagent cannot be easily adapted to commercial scale synthesis due to its high cost. In addition, there is an ongoing need to discover methods that provide higher yields, thereby improving production economics and / or reducing waste. Preferably, new methods should use less expensive starting materials.

[0006] There is a need for methods suitable for preparing larger quantities of Compound (I) than are typically prepared by laboratory-scale methods. There is also a need for methods that provide higher yields of Compound (I) than previously disclosed methods.

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

[0008] In one aspect, the present invention provides a method for preparing compound (I): It contains the following steps: 1) Compound 1 having the following structure With the following compound 2 in X is selected from Cl, Br and I; Y is selected from OR 9 NHOC 1-3 alkyl, Cl, Br and I; and R 9 Selected from C 1-3 Alkyl, C 1-3 Hydroxyalkyl, substituted phenyl and substituted benzyl; react in a suitable solvent to generate the following compound 3a or 3b 2) In the presence of an acid, convert compound 3a or 3b into the following compound 4 3) Compound 4 is then reacted with trialkyl orthoformate in an alcohol solvent to generate compound 5 of the following formula: in R 1 It is C 1-6 alkyl; And R 3 'Selected from C 1-6 alkyl, optionally substituted phenyl and benzyl; 4) Its ester hydrolysis under alkaline conditions or when R 3 When ' is a substituted benzyl group, hydrogenation is performed to form the following compound 6 Where R 1 and X is as defined above; 5) The carboxylic acid portion of compound 6 is then activated and reacted with a chiral auxiliary to form compound 6a of the following formula: Where Xa is a chiral auxiliary and R 1 and X is as defined above; 6) Compound 6a is then reacted with a base in the presence of a chiral auxiliary and a methyl donor such as a halogenated alkyl to form compound 7 Where R 1 , X and Xa are as defined above; 7) Remove the chiral auxiliary Xa to obtain the following compound 7 Where R 1 and X is as defined above; 8) Then, in the presence of a metal catalyst, compound 7 is reacted with compound 8 having the following structure: To generate compound 9 9) Reducing the nitro group in compound 9 to form the following compound 10 10) Cyclization of compound 10 using a suitable coupling agent to generate compound 11 11) In the presence of an acid, the keto functional group is unmasked to generate compound 12 12a) using an ammonia equivalent in the presence of a reducing agent or 12b) using a transaminase in the presence of an ammonia source, a different recycling system and a cofactor to reduce compound 12 to generate the amine stereocenter present in compound 13 13) which is then coupled with compound 14 of the following structure To generate compound (I):

[0009] In yet another aspect, the present invention provides a compound of formula (II): or in the form of a free base or a salt, wherein ---- is an optional bond, R 1 It is C 1-6 Alkyl, preferably Me; R 2 It is C 1-3 Alkyl, preferably Me; R 3 Selected from OH, OC 1-4 alkyl, in R 6 Selected from C 1-3 Alkyl, phenyl and benzyl; R 7 Selected from H and phenyl; R 8 Selected from C 1-3 alkyl, phenyl and benzyl; and X is selected from F, Cl, Br and I.

[0010] In some embodiments of the compound of formula (II): R 1 It is C 1-6 alkyl; R 2 It is C 1-3 alkyl; R 3 Selected from OH, OC 1-6 alkyl, in R 6 Selected from C 1-3 Alkyl, phenyl and benzyl; R 7 Selected from H and phenyl; R 8 Selected from C 1-3 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 pharmaceutically acceptable salt thereof: R 1 It is methyl; R 2 It is methyl; R 3 is OH; and X is Cl.

[0012] In some embodiments of the compound of formula (II): R 1 It is methyl; R 2 It is methyl; R 3 is OH; and X is Cl; the compound is in the form of its free base or its (1S,2R)-2-amino-1,2-diphenylethane-1-olate or its dicyclohexylamine salt.

[0013] In yet another aspect, the present invention provides a compound of formula (IIa): or in the form of a free base or a salt, wherein ---- is an optional bond, R 1 It is C 1-6 Alkyl, preferably Me; R 2 It is C 1-3 Alkyl or C 1-3 Alkenyl, preferably Me or CH2; R 3 Selected from OH, OC 1-4 alkyl, in R 6 Selected from C 1-3 Alkyl, phenyl and benzyl; R 7 Selected from H and phenyl; R 8 Selected from C 1-3 alkyl, phenyl and benzyl; and X is selected from F, Cl, Br and I.

[0014] In some embodiments, the method of making a compound of formula (II): in R 1 It is methyl; R 2 It is methyl; R 3 is OH; and X is Cl; The method comprises the following steps: a) making the following compound 22 With the following compound 2 in X is selected from Cl, Br and I; Y is selected from OR 9 NHOC 1-3 alkyl, Cl, Br and I; and R 9 Selected from C 1-3 Alkyl, C 1-3 Hydroxyalkyl, substituted phenyl and substituted benzyl; react in a suitable solvent to generate the following compound 23 b) in the presence of an acid, converting compound 23a or 23b into compound 24 of the following formula c) Compound 24 is then reacted with trimethyl orthoformate or triethyl orthoformate to produce compound 25 of the following formula Where R 1 is methyl or ethyl, and X is as defined above; d) using an enzyme to convert compound 25 into compound 26 of the following formula Where R 1 and X is as defined above; e) hydrolyzing compound 26 to form compound 27 Where R 1 and X is as defined above.

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

[0016] In some embodiments, the method of preparing a compound of formula (II) or formula (IIa) having the structure of compound 21: The method comprises the following steps: 1) Make the following cyclopentane ester derivative Where R is C 1-6 alkyl, Reaction with dialkylamine to produce the following compound 40 Where Alk is C 1-6 alkyl; 2) Compound 40 is combined with a first base to form compound 41 having the following structure: 3) Compound 41 is reacted with an acid and an alcohol R 3 'OH reacts to form the following compound 40 Where R 3 'It's C 1-6 alkyl; 4) reacting compound 42 with compound 2a having the following structure: The reaction is carried out in the presence of a second base to produce the following compound 18b 5) Compound 18b is converted into compound 20b having the following structure and 6) Hydrogenating compound 20b to generate a compound of formula (II) having the following structure

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

[0018] In some embodiments, the method of preparing a compound of formula (II) or formula (IIa) having the chemical formula of compound 19: in R1 is C 1-6 alkyl; R 10 It is C 1-6 alkyl; The following steps are involved: 1) Compound 37 of the following formula: Reaction with pyruvate phosphonium alkane to form compound 37: Where R 3 'Independently is C 1-6 alkyl; 2) Compound 38 is reacted with compound 2a having the following structure: In the presence of a base, the reaction is carried out to form the following compound 39: 3) treating compound 39 with a first acid to form compound 18a with the following structure: Compound 18a was reacted with C 1-6 The alkyl alcohol and the second acid and optionally a drying agent are reacted to form compound 19.

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

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

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

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

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

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

[0025] In yet another aspect, the present invention provides a compound of formula (VI): or in the form of a free base or a salt, wherein R 2 It is C 1-3 alkyl; R 3 Selected from OH, OC 1-4 alkyl, and in R 6 Selected from C 1-3 Alkyl, phenyl and benzyl; R 7 Selected from H and phenyl; R 8 Selected from C 1-3 alkyl, phenyl and benzyl; and X is selected from F, Cl, Br and I.

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

[0027] In some embodiments, the compound of formula (VI) has the structure of compound 34:

[0028] In another aspect, the present invention provides a method for treating a thrombotic disorder comprising administering to a mammal, preferably a human, in need thereof a therapeutically effective amount of Compound (I), wherein Compound (I) is prepared using the novel process steps of the present invention. DETAILED DESCRIPTION definition

[0029] Unless otherwise indicated, as used herein, the term "alkyl" refers to a straight or branched chain, saturated aliphatic group containing one to ten carbon atoms, for example, alkyl groups including methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, and the like. The term "lower alkyl" refers to an alkyl group having one to four carbon atoms.

[0030] The term "alkoxy" refers to a group having the formula -O-alkyl, wherein the alkyl group as defined above is attached to the parent molecule via an oxygen atom. The alkyl portion of the alkoxy group may have 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), tert-butoxy (-OC(CH3)3 or -OtBu), and the like.

[0031] The term "aryl" refers to a monocyclic or fused bicyclic ring assembly containing 6 to 10 ring carbon atoms, wherein each ring is aromatic, for example, phenyl or naphthyl.

[0032] The term "substituent" refers to additional substituent groups selected from halogen (preferably fluorine, chlorine or bromine), hydroxy, amino, mercapto and the like. Preferred substituents for the groups described herein as substituted lower alkyl or substituted alkyl are halogen, preferably fluorine substituents.

[0033] The term "reducing agent" refers to any reagent that reduces the oxidation state of a carbon atom in the starting material by adding a hydrogen atom to such carbon or adding an electron to such carbon, and thus should be apparent to one of ordinary skill and expertise in the art. The definition of "reducing agent" includes, but is not limited to, borane-dimethyl sulfide 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, hydrogen Ni / B, and similar reagents. Certain acidic and Lewis acidic reagents enhance the activity of the reducing agent. Examples of such acidic reagents include acetic acid, methanesulfonic acid, hydrochloric acid, and the like. Examples of such Lewis acidic reagents include trimethoxyborane, triethoxyborane, aluminum chloride, lithium chloride, vanadium trichloride, dicyclopentadienyltitanium dichloride, cesium fluoride, potassium fluoride, zinc (II) chloride, zinc (II) bromide, zinc (II) iodide, and the like.

[0034] The term "removable protecting group" or "protecting group" refers to any group that, when bound to a functional group such as an oxygen atom of a hydroxyl or carboxyl group or a nitrogen atom of an amino group, prevents reactions from proceeding at such functional groups, and which can be removed by conventional chemical or enzymatic procedures to reconstruct the functional group. The specific removable protecting group employed is not critical.

[0035] As used herein, the term "ligand" refers to a phosphine derivative attached to palladium, such as a monodentate or bidentate aryl or alkyl phosphine, which is capable of complexing the palladium atom. The term is well known to those skilled in the art.

[0036] As used herein, the term "silylation" or "silylating" refers to a method of introducing an alkenyl or silicon-containing group. Silyl groups include, but are 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 "desiliconization" refers to a process of removing silane groups or silicon-containing groups. Embodiments of the present invention

[0038] The present invention relates to certain synthetic intermediates and processes for preparing those intermediates and compounds (I).

[0039] General aspects of these exemplary methods are described in the Schemes and Examples.Each of the products of the following methods are optionally separated, isolated and / or purified prior to their use in subsequent methods.

[0040] In general, reaction conditions such as temperature, reaction time, solvent, workup procedures and the like should be those common in the field of the particular reaction to be performed. In general, the temperature should be from -100°C to 200°C, the solvent should be aprotic or protic, and the reaction time should be from 10 seconds to 10 days. The workup usually consists of quenching any unreacted reagents, followed by partitioning the aqueous / organic layer system (extraction) and separating the layer containing the product.

[0041] Typically, the oxidation and reduction reactions are carried out at temperatures close to room temperature (about 20° C.), but for metal hydride reductions the temperature is often lowered to 0° C. to −100° C. The solvent used for the reduction is usually aprotic, and the solvent used for the oxidation may be protic or aprotic. The reaction time is adjusted to achieve the desired conversion.

[0042] In one embodiment, the present invention provides a method for preparing compound (I). Representative general methods for preparing derivatives are summarized in Schemes 1 and 2 below. Solution 1

[0043] Each step of the preparation method illustrated in the above scheme will now be described in more detail.

[0044] Step 1

[0045] The starting materials for this method are compound 1 and compound 2. For those embodiments in which the starting materials are prepared according to literature methods, the starting materials are preferably purified before the reaction. Compound 1 and compound 2 are reacted under basic conditions in a suitable solvent to form compound 3. + 、Na + and K + Alkoxide bases having a counter cation of 2-(4-butoxide)-4-ol are suitable, such as methoxide, ethoxide, tert-butoxide, pentoxide, tert-pentoxide.

[0046] Examples of suitable solvents include, but are not limited to, polar aprotic solvents such as dimethyl orthoformate, dimethyl sulfoxide, and N-methylpyrrolidone; ether solvents such as tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), methyl tert-butyl ether (MTBE), diethoxymethane, and (CPME); hydrocarbons such as benzene, toluene, hexane, and heptane; halogenated solvents such as dichloromethane and 1,2-dichloroethane; acetates such as ethyl acetate, isopropyl acetate, and butyl acetate, and other solvents such as acetonitrile, methyl vinyl ketone, N,N-dimethylacetamide; polar aprotic solvents such as and mixtures thereof. Preferred solvents include ether solvents such as tetrahydrofuran, 2-methyltetrahydrofuran, and diethoxymethane.

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

[0048] Step 2

[0049] Compound 3 is then subjected to a retro-Claisen reaction under acidic conditions or aqueous acidic conditions to generate 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] The reaction temperature can vary over a relatively wide range. The reaction is usually carried out at a temperature of 0°C to 80°C. Preferably, the reaction is carried out at about 20°C to about 65°C.

[0051] Step 3

[0052] Compound 4 is then converted to its corresponding ester and ketal using an alcohol solvent, an acid catalyst, and an optional drying agent in the presence of a trialkyl orthoformate. 1-6 Alcohol solvents such as methanol, ethanol, propanol, butanol, pentanol and hexanol. Acid catalysts may be selected from HCl, trimethylsilyl chloride (TMSCl), pyridine p-toluenesulfonic acid (PPTS), p-toluenesulfonic acid (PTSA), a desiccant may be required and may be selected from Na2SO4 and MgSO4, and trialkyl orthoformate may be selected from, but not limited to, trimethyl orthoformate (TMOF) and triethyl orthoformate (TEOF).

[0053] Step 4

[0054] The ester in compound 5 is then hydrolyzed under basic conditions in the presence of water and a suitable organic solvent that is stable under basic conditions (such as toluene, NMP). A suitable base is a Li+ 、Na + , K + , Cs + or NH4 + Hydroxides as Countercations Non-limiting examples of hydroxides with counter anions are KOH, NaOH, and LiOH.

[0055] Step 5

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

[0057] Chiral auxiliary agents include, but are not limited to, oxazolidinone, 8-phenylmethanol, transphenylcyclohexane, camphorsulfone, pseudoephedrine (R,R) or (S,S) or pseudoephedrine amide (R,R) or (S,S), alkylthiazolidine-2-thione derivatives or N-(-3-hydroxy-4,7,7-trimethylbicyclo[2.2.1]heptan-2-yl)-N-phenylbenzenesulfonamide. In one embodiment, the chiral auxiliary agent is an oxazolidinone selected from the following

[0058] The base may be selected from, for example, DIPEA, TEA, LDA, n-BuLi, sec-BuLi or tert-BuLi, potassium tert-butoxide in a suitable solvent with or without an inorganic salt such as LiCl.

[0059] Step 6

[0060] Compound 6a is alkylated to form compound 6b using an alkylating agent and a strong base. Non-limiting examples of activators include alkyl halides, dialkyl sulfates, trialkyl tinium tetrahydroborate. Preferably, the alkylating agent is a methyl halide, such as MeI. Suitable bases are NaHMDS, LiHMDS, KHMDS, LDA. Solvents may be selected from ether solvents (THF, 2-Me-THF, MTBE, CPME), aromatic solvents (toluene) or polar aprotic solvents or combinations thereof. Subsequently, a caustic soda such as LiOH, NaOH and KOH is used to remove the chiral auxiliary under alkaline conditions to produce compound 7.

[0061] Step 7

[0062] Compound 7 is separated as an ammonia base or a basic salt of Na or K in a desired solvent. Suitable bases are dibenzylamine, DABCO, dicyclohexylamine, ethanolamine, diethanolamine, imidazole, arginine, lysine, trimethylamine, 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 can be separated 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 produce compound 9. The metal catalyst may be derived from Pd, Pt, Rh, Ru, Ir, Fe, Ni, or Cu. A ligand such as a phosphine (i.e., CX-A, XPhos, SPhos, Xantphos, DCEPhos) or an N-heterocyclic carbene (i.e., IMes, Ipr) may assist the reaction. Suitable bases include organic bases (i.e., Et3N, DIPEA), inorganic bases (i.e., KOPiv, KOAc, K2CO3), or bases derived from an inorganic base and a carboxylic acid (i.e., K2CO3 / PivOH, Cs2CO3 / PivOH, K2CO3 / PhCO2H). Suitable solvents are ether 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] Compound 9 is then subjected to a nitro reduction step using a metal catalyst such as Pd, Pt, Rh in the presence of hydrogen or a hydrogen transfer agent such as ammonium formate or sodium formate on a support such as charcoal, alumina in an ether solvent or an alcohol solvent to form compound 10. Compound 9 can also be subjected to HSiCl3 / DIPEA, SnCl2 or Na2S2O4 to produce compound 10.

[0067] Step 10

[0068] Compound 10 is then subjected to a macrolactamization step using a suitable carboxyl activator, a base in a suitable solvent. Suitable coupling agents are any of the well-known coupling agents for coupling ammonia with an acid to form an amide. Non-limiting examples of coupling agents include PyBOP, HATU / HOBt, EDAC, oxalyl chloride, anhydrides such as pivalic anhydride, acid chlorides such as pivalic anhydride, or activators such as DPPCL, DMC or TCFH. Suitable solvents are typically ether solvents (THF, 2-Me-THF, MTBE, CPME), aromatic solvents (toluene).

[0069] Step 11

[0070] In step 11, the keto functional group in compound 11 is demasked under aqueous acidic conditions to provide compound 12. Non-limiting examples of acids include HCl, HBr and TFA.

[0071] Step 12

[0072] Compound 12 is then subjected to a reductive amination step to provide compound 13. This transformation can be achieved using a reducing agent such as BH3, NaBH3CN, Pd / C, Pt / C, wherein if Pd / C or Pt / C is used, an ammonia donor such as ammonia or an ammonium salt such as ammonium chloride, a hydrogen transfer salt such as ammonium formate, or hydrogen gas is present.

[0073] Reductive amination can also be achieved using a transaminase in the presence of an ammonia source such as isopropylamine, alanine, 3-aminobutyric acid, and methylbenzylamine and in the presence of a cofactor such as PLP. For the latter, the preferred solvent is aqueous DMSO. Different recirculation systems can be used, such as transaminase / lactate dehydrogenase / glucose dehydrogenase and transaminase / amino acid dehydrogenase / formate dehydrogenase. Non-limiting examples of transaminases are ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256, and ATA-260.

[0074] The transaminase used in the methods of the present disclosure generally comprises 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 greater 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 transaminase is an engineered transaminase polypeptide comprising an amino acid sequence that has one or more amino acid residue differences 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 capable of hybridizing under highly stringent conditions encodes a transaminase polypeptide having the percentage identity and one or more residue differences described above 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 methods described herein, the aminotransferase uses an amino donor to form a product compound. In some embodiments, the amino donor in the reaction conditions comprises a compound selected from isopropylamine (also referred to herein as "IPM") or any other amino donor suitable for the reaction in question. In some embodiments, the amino donor is IPM.

[0076] Suitable reaction conditions for the methods also typically include the presence of a cofactor in the reaction mixture. Because transaminases typically use members of the vitamin B6 family, the reaction conditions may include a cofactor selected from the group consisting of: pyridoxal-5'-phosphate (also known as pyridoxal-phosphate, PLP, P5P), pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM) and its phosphorylated counterparts; pyridoxine phosphate (PNP) and pyridoxamine phosphate (PMP). In some embodiments, suitable reaction conditions may include the presence of a cofactor selected from the group consisting of: PLP, PN, PL, PM, PNP, and PMP. In some embodiments, the cofactor is PLP.

[0077] Step 13

[0078] Final compound 13 was coupled with compound 14 to produce compound (I) as described in WO 2015 / 116886.

[0079] In the above methods, additional steps may be employed in steps 1-13. In addition, different synthetic methods may be employed in Schemes 1 and 2 to prepare key intermediates. Scheme 3 shows different methods for preparing specific embodiments of compound 7 (Scheme 1) as compound 21. Solution 3

[0080] Compound 15 can be coupled with 3-chloro-1,1-dimethoxypropane, 3-bromo-1,1-dimethoxypropane or 3-iodo-1,1-dimethoxypropane in a suitable solvent such as THF in the presence of a metal such as Mg and in the presence of an initiator such as I2 to form compound 16. The ketal is then hydrolyzed using an organic acid such as TFA, MSA, BSA, PTSA, PPTS or an inorganic acid such as HCl, HBr in the presence of water and a suitable solvent to provide compound 17. The aldehyde in compound 17 is then reacted with a triphenylphosphonium alkane such as methyl 2-(triphenyl-15-phosphinyl)propionate or ethyl 2-(triphenyl-15-phosphinyl)propionate, or with a phosphonate derivative such as methyl 2-(diethoxyphosphinyl)propionate or ethyl 2-(diethoxyphosphinyl)propionate in a suitable solvent in the presence of a base such as NaH or KOtBu to generate compound 18. Then use C 1-6 The ketone in compound 18 is protected as its corresponding ketal compound 19 using an alcohol solvent and an acid such as HCl, trimethylsilyl chloride (TMSCl), pyridine p-toluenesulfonic acid (PPTS), p-toluenesulfonic acid PTSA as a catalyst and an optional drying agent such as Na2SO4, MgSO4, trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF). The ester is then hydrolyzed to compound 20 under basic conditions in the presence of water. A suitable base is a Li + 、Na + , K + , Cs + NH4 + Hydroxide as a counter cation. Finally, the olefin in compound 20 is reduced using a metal catalyst in the presence of H2 to generate compound 21. The metal is preferably Ru or Rh. The chirality at the methyl carbon center is induced by using a suitable chiral ligand. Alternatively, compound 19 can be reduced by treating an ene reductase and hydrolyzing the ester under alkaline conditions.

[0081] Compound 19 can be prepared by selective condensation (Scheme 4). Solution 4

[0082] Compound 38 can be formed by coupling aldehyde 37 with triphenylphosphonium alkane such as methyl 2-(triphenyl-15-phosphinyl)propionate or ethyl 2-(triphenyl-15-phosphinyl)propionate in a suitable solvent in the presence of a base such as NaH or KOtBu, or reacting with a phosphonate derivative such as methyl 2-(diethoxyphosphinyl)propionate or ethyl 2-(diethoxyphosphinyl)propionate to produce compound 38. The obtained diester 38 is reacted with compound 2 having the structure of compound 2a in the presence of a base such as LiHMDS, LDA, tBuOK in a suitable solvent to produce compound 39. This compound is further decarboxylated in the presence of an acid such as HCl, MSA, H3PO4 in a suitable solvent to produce compound 18a, which is then converted into compound 19 as described above.

[0083] In another embodiment, compound 21 can be prepared starting from a cyclopentyl ester derivative (Scheme 5). Solution 5

[0084] Compound 40 is formed by coupling the cyclopentyl ester derivative with formaldehyde and a dialkylamine. Further treatment under basic conditions provides the diacid derivative compound 41. After esterification to form compound 42, coupling with compound 2a and acidic treatment, the acrylate derivative compound 18b is obtained. Subsequent use of a cyclopentyl ester derivative such as C 1-6 The ketone in compound 18b is protected as its corresponding ketal using an alcohol solvent, an acid such as HCl, trimethylsilyl chloride (TMSCl), pyridine p-toluenesulfonic acid (PPTS), p-toluenesulfonic acid PTSA as a catalyst and an optional drying agent such as Na2SO4, MgSO4, trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF). The ester is then hydrolyzed to compound 20b under basic conditions in the presence of water. Suitable bases include those with Li + 、Na + , K + , Cs + NH4 + Hydroxide or free carboxylic acid as counter cation. Finally, metal catalytic reduction of the olefin in compound 20b is used in the presence of H2 to generate compound 21. The metal is preferably Ru or Rh. The chirality at the methyl carbon center is induced by using a suitable chiral ligand. Alternatively, the desired mirror image isomer can be obtained by treatment with ene reductase.

[0085] In another embodiment, specific examples of compound 7 (Scheme 1), compound 27 (Scheme 7) that can be prepared via an enzymatic resolution pathway are shown in reaction schemes 6 and 7. Solution 6

[0086] Compound 22 is reacted with compound 2 under basic conditions in a suitable solvent to form compound 23. For example, alkoxides (methoxide, ethoxide, tert-butoxide, pentanolate, tert-pentanolate) have Li + 、Na + , K + Bases as counter cations and solvents such as ether solvents (THF, 2-MeTHF, MTBE, CPME), aromatic solvents (toluene) or bipolar aprotic solvents are suitable. Compound 23 is subjected to a reverse Claisen reaction under basic conditions or aqueous acidic conditions to obtain compound 24. Suitable acids are (but not limited to) H2SO4, MSA, BSA, nitric acid, TFA or perchloric acid.

[0087] Compound 24 is then converted to its corresponding ester and ketal compound 25 using an alcohol solvent (C1-6 alcohol) and an acid such as (but not limited to) HCl, trimethylsilyl chloride (TMSCl), pyridine p-toluenesulfonic acid (PPTS), p-toluenesulfonic acid PTSA as a catalyst and an optional drying agent such as Na2SO4, MgSO4 and a trialkyl orthoformate such as trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF).

[0088] Compound 25 was then subjected to enzymatic resolution as shown in Scheme 7. Solution 7

[0089] The racemic compound 25 is subjected to an enzymatic resolution step. When the desired enantiomer compound 26 is not reacted, the undesired enantiomer in the racemic mixture compound 25 is hydrolyzed. Compound 26 is then hydrolyzed using an aqueous base such as NaOH to provide compound 27. Preferably, the hydrolase is lipase MH Amano 10SD, which shows good selectivity (>90% enantiomer excess).

[0090] In option B, the racemic compound 25 is subjected to an enzymatic resolution step. The desired enantiomer in the racemic mixture compound 25 is hydrolyzed to form compound 27 when the undesired enantiomer is not reacted to a certain extent.

[0091] In Option C, the unreacted undesired enantiomer compound S-26 generated in Option B is racemized in the presence of a base to form Compound 25 which can be used as a starting material in Option A or Option B above.

[0092] In another embodiment, intermediate compound 10 is prepared by the method shown in Scheme 8, which is different from the method in Scheme 1. Solution 8

[0093] Step 1

[0094] Compound 28 is coupled with silyl protected acetylene in a strong base and a suitable solvent to produce compound 29. The base may be a strong lithiating base, such as an alkyl lithiating base or an aryl lithiating base. Non-limiting examples of alkyl and aryl lithiating bases are methyl lithium, n-butyl lithium, sec-butyl lithium, tert-butyl lithium and phenyl lithium. The solvent may be an ether solvent, such as THF.

[0095] Step 2 Subsequently, an alcohol solvent (C 1-6 The ketone moiety is protected as its corresponding ketal using an alcohol) and an acid such as HCl, trimethylsilyl chloride (TMSCl), pyridine p-toluenesulfonic acid (PPTS), p-toluenesulfonic acid PTSA as a catalyst and an optional drying agent such as Na2SO4, MgSO4 and an orthoformate trialkyl ester such as trimethyl orthoformate (TMOF) or triethyl orthoformate (TEOF). The protective silylation group is then deprotected using a fluorine source such as TBAF, HF.TEA, HF in a suitable solvent such as THF, 2-MeTHF to generate compound 30.

[0096] Step 3

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

[0098] Step 4

[0099] Compound 31 is then subjected to metal-catalyzed staggered coupling with commercially available (S)-(-)-3-methoxy-2-methyl-3-oxopropylzinc bromide to produce compound 32. Non-limiting examples of metal catalysts include Pd(II) salts such as PdCl2, Pd(OAc)2, or pre-attached metals such as 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride.

[0100] Step 5

[0101] Compound 32 is then reacted with compound 8 in the presence of a metal catalyst and a base to produce compound 33. The metal catalyst may be derived from Pd, Pt, Rh, Ru, Ir, Fe, Ni, or Cu. A ligand such as a phosphine (i.e., CX-A, XPhos, SPhos, Xantphos, DCEPhos) or an N-heterocyclic carbene (i.e., IMes, Ipr) may assist the reaction. Suitable bases include organic bases (i.e., Et3N, DIPEA), inorganic bases (i.e., KOPiv, KOAc, K2CO3), or bases derived from an inorganic base and a carboxylic acid (i.e., K2CO3 / PivOH, Cs2CO3 / PivOH, K2CO3 / PhCO2H). Suitable solvents are ether 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 enable reduction of the double bond and nitro functionality, and the methyl ester is then hydrolyzed to afford compound 10. Reduction using metals such as Pd or Pt in the presence of hydrogen in protic agents such as MeOH, EtOH, IPA can be effective. Ester hydrolysis is performed by treating the methyl ester with a base hydroxide such as LiOH, NaOH, KOH in the presence of water or water and a miscible organic solvent. Solution 9

[0104] Step 1

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

[0106] Steps 2 and 3

[0107] Compound 46 can be prepared by azidation of the compound using a suitable alkyne followed by click chemistry. Compound 45 is subjected to azidation conditions such as TMSN3 / tBuONO to provide an intermediate azide, which is then treated with trimethylsilyl acetylene in the presence of a copper (I) catalyst such as CuOAc or copper (I) iodide to produce triazole compound 46.

[0108] Step 4

[0109] Compound 47 can be made from silanyl compound 46 by reaction with 1,3-dichloro-5,5-dimethylhydantoin in a suitable solvent. Suitable solvents include polar aprotic solvents such as THF or DMF.

[0110] Step 5

[0111] Compound 14 can be prepared from compound 47 by reaction in hydrochloric acid, such as concentrated hydrochloric acid.

[0112] In another embodiment, the present invention provides a compound of formula (II): It is in the form of a base or a salt, wherein ---- is an optional bond, R 1 It is C 1-6 alkyl; R 2 It is C 1-3 Alkyl; Alkenyl R 3 Selected from OH, OC 1-6 alkyl, and in R 6 Selected from C 1-3 Alkyl, phenyl and benzyl; R 7 Selected from H and phenyl; R 8 Selected from C 1-3 alkyl, phenyl and benzyl; and X is selected from F, Cl, Br and I.

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

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

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

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

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

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

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

[0120] In another embodiment, the present invention provides a compound of formula (V): in R 2 It is C 1-3 alkyl; and R 5 Selected from CHF2, CD3 and CH3.

[0121] In another embodiment, the present invention provides a compound

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

[0123] In another embodiment, the present invention provides a compound having the following structure. Example

[0124] In order to better illustrate the present invention, the following examples are provided. Unless otherwise indicated, all reactions were performed under a nitrogen atmosphere using dehydration techniques. Unless otherwise indicated, reagents were used when received from suppliers. The yields cited are for isolated materials and are not corrected for water content. CH3CN / H2O / MeOH was used as mobile phase (containing 0.05% TFA or 0.1% NH4OAc) on a Shimadzu system by normal or reverse phase HPLC monitoring reactions.

[0125] Method A

[0126] Chromatographic conditions

[0127] gradient Time (minutes) ％A ％B 0 100 0 6 90 10 11 10 90 12.5 10 90 12.6 100 0 15 100 0

[0128] Method B

[0129] Chromatographic conditions

[0130] gradient Time (minutes) ％A ％B 0 100 0 10 0 100 15 100 0

[0131] Method C

[0132] Chromatographic conditions

[0133] gradient

[0134] Method D

[0135] Chromatographic conditions

[0136] gradient

[0137] Method E

[0138] Chromatographic conditions String Lux Cellulose-3, 4.6×150mm, 3 microns Column temperature 25℃ Flow rate 0.8 ml / min Injection volume 10μL wavelength 220nm Mobile Phase A 0.05% TFA in water: CH3CN (95:5) Mobile Phase B 0.05% TFA in water: CH3CN (5:95)

[0139] gradient:

[0140] Method F

[0141] Chromatographic conditions String Ascentis Express C18 2.7um 4.6×150mm Column temperature 35℃ Flow rate 0.8 ml / min Injection volume 5-10μL wavelength 265nm Mobile Phase A 0.05% Formic acid in MeOH:water (20:80) Mobile Phase B 0.05% Formic acid in ACN:MeOH (80:20)

[0142] gradient:

[0143] Method G

[0144] Chromatographic conditions String Zorbax Eclipse Plus C8 1.8um 4.6×50mm Column temperature 25℃ Flow rate 1.2 ml / min Injection volume 5-10μL wavelength 220nm Mobile Phase A 0.05% TFA in water: CH3CN (95:5) Mobile Phase B 0.05% TFA in water: CH3CN (5:95)

[0145] gradient:

[0146] Method H

[0147] Chromatographic conditions String Chiralpak IG-3, 4.6×150mm, 3um Column temperature 30℃ Flow rate 1.0 ml / min Injection volume 10μL wavelength 220nm Mobile Phase A 0.1% DEA in heptane Mobile Phase B 0.1% DEA in IPA

[0148] gradient:

[0149] Method I

[0150] Chromatographic conditions

[0151] gradient: Time (minutes) ％A ％B 1 0.0 90 10 2 10 70 30 3 15 60 40 4 20 50 50 5 30 50 50 6 40 10 90

[0152] Method J

[0153] Chromatographic conditions-

[0154] gradient: Time (minutes) ％A ％B 1 0 80 20 2 10 0 100

[0155] Method K

[0156] Chromatographic conditions-

[0157] gradient: Time (minutes) ％A ％B 1 0 60 40 2 15 60 40

[0158] Method L

[0159] Chromatographic conditions-

[0160] gradient: Time (minutes) ％A ％B 1 0 90 10 2 15 90 10 3 18 0 100 4 22 0 100

[0161] NMR spectra were recorded on a Bruker DRX-600, DRX-500 or DRX 400 instrument and referenced to residual undeuterated solvent. Low resolution mass spectra (LRMS) were recorded on a Water ZQ ES instrument.

[0162] As used herein, abbreviations are defined as follows: "1×" is once, "2×" is twice, "3×" 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 normal, "M" is molar, "mmol" is millimole, "min" is minute, "h" is hour, "rt" is room temperature, "RT" is residence time, "atm" is atmospheric pressure, "psi" is pounds per square inch, "conc." is concentration, "sat" or "sat'd" is saturated, "MW" is molecular weight, "mp" is melting point, "ee" is enantiomer 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 spectrometry, "HPLC" is high pressure liquid chromatography, "RPMS" is liquid chromatography mass spectrometry, "HPLC" is reverse phase HPLC, "TLC" or "tlc" is thin layer chromatography, "NMR" is nuclear magnetic resonance spectroscopy, "nOe" is nuclear Overhauser effect spectroscopy, " 1H" is a proton, "δ" is Δ, "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 stereochemical symbols familiar to those skilled in the art.

[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. Embodiments of General Scheme 2 for forming compounds (I) are described in Examples 5-12 below. Solution 10 Example 1 Synthesis of compound 3a-Cl

[0164] Compound 2a (540 g, 3053 mmol, limited reagent) was loaded into a 20 L reactor equipped with a nitrogen inlet, a temperature probe, and an overhead stirrer, followed by THF (4500 mL), and stirring was started to achieve dissolution of compound 2a. Compound 1 (287.4 g, 3382 mmol, 1.2 eq) was then added, followed by washing with THF (50 mL).

[0165] The solution was cooled to -35°C using a chiller. Potassium tert-butoxide (1 M, 3650 mL, 1.3 eq) was then added in THF solution at a rate such that the temperature did not exceed -30°C.

[0166] The reaction mass was kept at -35 to -30 ° C for 1 hour until the reaction was completed. Water (3635 mL) was loaded into another 20 L reactor, followed by the addition of concentrated sulfuric acid (193.3 g, 0.69 eq). The reactor jacket temperature was set to 10 ° C, and the batch was cooled to 12 ° C. The cooling solution from the freezing reactor (-35 to -30 ° C) was loaded into a 20 L reactor containing cooled aqueous H2SO4 via a delivery pipe, keeping the temperature <10 ° C. After completing the delivery, THF was then distilled out to ~7.5 L volume at 20 to 25 ° C under vacuum. Solids were now formed. The slurry was filtered and washed with water (2000 mL, 3.7 V). 1,061 g of compound 3a-Cl in the form of a yellow-brown solid was obtained.

[0167] 1 H NMR(500MHz,DMSO-d6)δ14.97(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 + [M+H] + The LRMS calculated value is 224.05 and the observed value is 224.28. Example 2 Synthesis of compound 4a

[0169] 70wt%MSA solution (1074g, 7823mmol, 2.83eq) is loaded into a 20L freezing reactor, followed by water (4900mL). The reaction mixture is heated to 65°C. The wet cake of compound 3a-Cl is then loaded into the reactor, and the reaction mass is heated at 65°C for 3 hours until termination. The reaction mass is then cooled to 20-25°C, and NH4OH (28wt%) aqueous solution (489g, 3907mmol, 1.41eq) is added. pH is adjusted to 5.06 using a pH probe. The resulting slurries are heated to 44°C, and kept all night at 44°C. The reaction mass is cooled to 20-25°C, and the slurries are filtered, and the resulting filter cake is washed with water (3000mL, 6V). The wet cake is dried 2 days using a vacuum oven (50°C, 100mmHg) to obtain 590g of compound 4a as a white solid.

[0170] 1 H NMR (500MHz, DMSO-d6): δ12.01(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 13ClNO3 + [M+H] + The LRMS calculated value is 242.06 and the observed value is 242.24. Example 3 Synthesis of compounds 5a and 6-Cl

[0172] MeOH (4 L) and compound 4a (367.3 g, 1500 mmol, 98.9 mass) were loaded into a 20 L reactor. 3 L of MeOH was then added. Subsequently, trimethyl orthoformate (734 mL, 6700 mmol, 100 mass%, 4.5 eq.) was added, followed by washing with MeOH (400 mL). Trimethylsilyl chloride (367 mL, 2880 mmol, 100 mass%, 1.92 eq.) was loaded, followed by addition of MeOH (200 mL). The reactants were heated to 49 °C internal temperature for 12 hours.

[0173] NaOH (10N) 1220mL is added to another 20L reactor, followed by 1620mL H2O, and the reaction mass is cooled to 0°C. The reactant contents containing compound 5a are transferred to the reactant containing aqueous NaOH. The internal temperature is raised from 5°C to 22°C. 700mL MeOH is used to clean the main reactor, and the contents are transferred to the quenching reactor. The reactant is stirred for 4 hours. The reactor is warmed to a jacket temperature of 20°C and stirred. MTBE (2570mL) is then loaded. Stirring is stopped and the aqueous product enriched layer is collected and continued to advance. 2985mL of 20wt% citric acid is then added to the stirred aqueous phase. When a pH of 5.3 is reached, a slurry is formed, and then filtered. 387.1g of compound 6-Cl (89.6% yield) in a solid state is obtained.

[0174] 1 H NMR(400MHz,DMSO-d6)δ11.98(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 + [M-CH3O] +The LRMS calculated value is 256.07 and the observed value is 256.24. Example 4 Synthesis of compound 7a-DCHA

[0176] Compound 6a (87.88 g, 305 mmol, limited reagent) was loaded into a 2L chemical glass reactor, followed by anhydrous THF (1760 mL). THF was distilled off, down to 10 Vol. The KF of the solution was <200 ppm. Additional THF (880 mL) and triethylamine, CAS 121-44-8 (106.4 mL, 2.5 eq) were added. The solution was cooled to 0 ° C, and trimethylacetyl chloride, CAS 3282-30-2 (44.13 g, 1.2 eq) was added via an addition funnel at a rate that the temperature did not exceed 5 ° C. After keeping for 30 minutes, lithium chloride (16.16 g, 1.2 eq) was added. After aging for 15 minutes, a solid chiral auxiliary, CAS 102049-44-7 (64.86 g, 1.2 eq) was loaded once. The slurry was allowed to warm to 20 ° C within 3 hours and matured overnight. THF was then distilled off under vacuum to a final volume of 800 mL. Toluene (530 mL) was loaded 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 phase was discarded. The organic phase was washed with 7 wt% NaHCO3 (270 mL) and water (270 mL). After phase separation, the lower aqueous phase was discarded. The organic layer was distilled down to 220 mL. And then anhydrous THF (1860 mL) was added. The solution was passed through a 0.45 micron fine filter.

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

[0178] THF (880 mL) was added and the solution was cooled to 0°C. A 30 wt% H2O2 solution (64.18 g, 1.82 eq) was then added, followed by a lithium hydroxide solution (12.42 g in 110 mL of water) over 10 minutes. After 6 hours, a 10 wt% sodium sulfite solution (63.48 g, 2.0 eq in 580 mL of water) was obtained. The mixture was aged for 1 hour. THF was then removed by distillation until ~700 mL was collected.

[0179] The pH was then adjusted to about 9.5 using 10N NaOH. Toluene (540 mL) was added. The biphasic mixture was mixed for 15 minutes and then stopped. The separated organic layer was further extracted with 360 mL of saturated NaHCO. The combined aqueous layer was overloaded in the reactor and extracted with MTBE (720 mL). The organic layer was discarded.

[0180] The reactor was reloaded with the product rich aqueous layer containing compound 7a, and MTBE (900 mL) was added. The pH was adjusted to 4.4 using citric acid.

[0181] MTBE was removed by distillation and replaced with MeCN / MTBE 4:1 (6 volumes) based on the input of compound 7a (valence corrected). The resulting liquid stream was finely filtered, and the solution was filtered with a fine filter. A portion of dicyclohexylamine (up to 1.5eq relative to compound 7a) was subsequently added. The slurry was heated to 55 ° C and held for 30 minutes. The reaction mass was then cooled to 0 ° C. The slurry was filtered on a Buchner funnel under N2 protection, washed with 2.0 volumes of cooling MeCN (0 ° C), dried under vacuum, and then kept in a vacuum oven at 50 ° C for 24 hours. 61g of compound 7a-DCHA (88.2% in the salt formation step, 61.6% overall yield from compound 6a in four steps) was obtained as a white solid. Compound 7 is a 1:1.5 complex of compound 7: dicyclohexylamine.

[0182] 1 H 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 + [M-CH3O] + The LRMS calculated value is 270.09 and the observed value is 270.24. Example 5 Synthesis of compound 9a

[0184] 2MeTHF (500 mL), catalyst [Pd(allyl)Cl]2 (1.57 g, 0.05 eq.) and Xphos (4.48 g, 0.055 eq.) were sequentially loaded into a 2L clean reactor equipped with an overhead stirrer, a thermocouple and a nitrogen inlet to produce a roughly homogeneous light yellow solution at 20°C. Salt compound 7a-DCHA (98.2 g, 1.0 eq.) and pyrrole compound 8 (32.3 g, 1.15 eq.) were added portionwise to the reactor to produce a white suspension. After 30 minutes, a portion of KOPiv (32.3 g, 1.3 eq.) was added to the solution, and the reactor was rinsed with 2-MeTHF (500 mL) bubbled with N2 for 30 minutes. Under N2, the solution was refluxed for 10 hours to produce a black suspension. The crude product was cooled to 20° C. and quenched with K 3 PO 4 (550 mL, 20% aq.) to a pH between 10.0 and 10.5, the aqueous layer was separated, and the organic layer was washed with K 3 PO 4 -K 2 HPO 4 buffer solution (800 mL, pH 10.2, aq.). The aqueous layers were combined and filtered to produce a black solution. 2-MeTHF (1300 mL) and activated carbon (13.9 g, Darco G-60) were added to the aqueous solution, and the solution was acidified to pH 5-6 using citric acid (254 g, 3.4 eq.) portionwise over 30 minutes. The suspension was stirred at 20° C. for 30 minutes. The suspension was filtered, and the organic layer was retained and concentrated under vacuum to 300 mL, and the solvent was exchanged with nBuOH (1000 mL) at 80° C. under 150 mbar vacuum. The concentration of the resulting solution was adjusted to 170-180 mg / mL (500 mL, no more than 5% wt 2-MeTHF in solution). This solution was gradually cooled to 0°C over 10 hours and maintained at 0°C for an additional 10 hours to produce a white slurry. The slurry was filtered through a Nutsche filter and the reactor was rinsed with nBuOH (100 mL) and the filter cake was washed with the resulting suspension. The filter cake was washed with heptane (100 mL) and dried in an oven (house vacuum 50°C for 24 hours). Isolated compound 9a (73.2 g, 95 wt%) was obtained in 83% yield.

[0185] 1H NMR (400MHz, DMSO-d6): δ8.89-8.80(m,1H),8.72-8.65(m,1H),7.88-7.52(m,3H),3.10-3.00(m,6H),2.50(dt,J=3.5,1.8 Hz,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)δ177.3,158.9,149.5,139.3,138.3,135.0,132.8,123.5, 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

[0187] THF (900 mL), Pd / C (4.6 g, 10% wt, 0.1 eq.) and compound 9a (46.0 g, 1.0 eq.) were sequentially loaded into a pressure reactor equipped with an overhead stirrer, a thermocouple and a nitrogen inlet to generate a suspension at 20° C. The reactor was swept with N2 and H2 three times each. The solution was stirred vigorously under H2 (40 psi) for 18 hours. The crude product was discharged from the reactor and filtered through a Nutsche filter. The THF solution was concentrated to a clear oily compound 10a (101 g, 40% wt) with a yield of 95%. A small sample was taken and thoroughly concentrated for spectral analysis.

[0188] 1H NMR (500MHz, CDCl3): δ8.77(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

[0190] 600 mL of THF was added to a 1 L reactor, followed by chloro-N, N, N', N'-tetramethylformamidine hexafluorophosphate (TCFH, 17.1 g, 59.7 mmol, 98.0 mass%, 1.52 eq). An additional 200 mL of THF was used to purge all TCFH in the reactor. N, N-diisopropylethylamine (14.6 mL, 83.7 mmol, 100 mass%, 2.13 eq) was added to the suspension and stirred at room temperature. A solution of compound 10a in THF (44.9 g, 39.2 mmol, 34.8 mass%, 1.00 eq) was loaded into a 50 mL syringe, and the solution was added to the reactor over 10 hours using a syringe-pump (ca. 5 ml / hour rate).

[0191] The solvent of the reaction stream (~50 mL, equivalent to 1 g of compound 11a input) was replaced from THF to MIBK (~20 mL). The organic layer was washed with aq. K2HPO4 (15%, 15 mL), followed by the addition of 1.2 eq (mol / mol with input Compound 11a) of solid fumaric acid. The mixture was then concentrated to ~8 mL under vacuum. A filtered tan product slurry was formed, the filter cake was washed with MIBK (2 mL), and then washed with heptane (2 mL), and dried under vacuum. Yield = 1.01 g (75.3% corrected for compound 11a input titer). Titer = 74.2 wt%, ee = 97.3%.

[0192] 1H NMR (400MHz, DMSO-d6) δ13.31-13.02(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(brd,J=7.1Hz,4H),0.46(br s,1H)

[0193] C 17 H 19 F2N4O2 + [M-CH3O] + The LRMS calculated value is 349.15 and the observed value is 349.08. Example 8 Synthesis of compound 12

[0194] Cyclopentyl methyl ether (2.5 mL) was added to a slurry of compound 11a-FUM (0.50 g, 67 wt %, 96 ee%) in water (5 mL), followed by trifluoroacetic acid (0.23 mL, 3.5 eq). The resulting mixture was heated to 45 ° C for 5 hours. The mixture was then cooled to ambient temperature and filtered. The reactor was washed with water (2.5 mL) and the washed filter cake was applied to wash. The filtrate was combined and the phases were separated. The resulting organic phase was extracted with aqueous HCl (0.5 N, 2.0 mL). The acidic aqueous extract was combined with the acidic aqueous phase previously 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 hours and filtered. The filtered filter cake was washed with water (5 mL×2) and MTBE (5 mL×2) and dried under vacuum to provide compound 12 (0.25 g, 78%, 97.2 ee%).

[0195] 1H NMR (500MHz, CDCl3): δ8.89(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 Alternatively, compound 12

[0197] Compound 9a (75.0 g 169 mmol), THF (525 mL) and Pt / V / C (~50% wet, 10 wt%, 7.5 g) were added to a pressurized 1 L container. The reactor was swept with N2 and H2 three times each. The solution was stirred vigorously at 20°C for 1 hour under H2 (1.5 bar) and then warmed to 40°C for 16 hours. The mixture was discharged from the reactor and filtered, the resulting solution was concentrated, and water was distilled off azeotropically by continuous distillation with THF. THF (1.2 L) was added continuously to another reactor, followed by the addition of chloro-N,N,N',N'-tetramethylformamidine hexafluorophosphate (TCFH, 70 g, 1.52 eq.) and N,N-diisopropylamine (103 mL, 3.5 eq.). The mixture was stirred vigorously and heated to 55°C. To this mixture was added a solution of compound 10 in THF (~50 mL / h) over 20 hours. The mixture was then concentrated, and a solvent conversion to acetonitrile was performed to achieve about 4L / kg of compound 11 in acetonitrile. Subsequently, an aqueous hydrochloric acid solution (3N, 23mL) was loaded, and the reaction mixture was heated at 55°C for 15 hours to form crude compound 22. The reactant was cooled to 10°C, and then 270mL of dichloromethane, 540mL of water, and hydrochloric acid (10.8N, 71mL) were loaded. After stirring for 1 hour, the phases were separated and the bottom organic layer was discarded. Potassium hydroxide (22.5wt%, ~169mL) was added to the aqueous layer at 10°C until the pH was increased to 3.0. After stirring for 1 hour at pH 3, potassium hydroxide (22.5wt%, ~17mL) was further added to the reactant until the pH reached 9.5. Compound 12 was isolated by filtration, washed with 169mL of water and 114mL of EtOH, and then dried. Example 10 Synthesis of compound 13

[0198] During the reaction, a 125 mL jacketed reactor with a water circulator was used to maintain a reaction temperature of 35 ° C. The pH was controlled at 7.5 using a calibrated pH steady state and 4 M aqueous isopropyl ammonia was loaded into the reactor. 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 loaded into the reactor. 1 M hydrochloric acid isopropyl ammonia solution was prepared in water, and 63 mL (final concentration 0.7 M) was loaded into the reactor. The reaction mixture was stirred for 2.0 minutes. The reactants were homogeneous. Aminase ATA-237 (0.5 g) was dissolved in 4.0 mL of 1 M hydrochloric acid isopropyl ammonia solution and added to the reactor. The enzyme container was cleaned with 3.0 ml of 1 M hydrochloric acid isopropyl ammonia solution and loaded into the same reactor. A 20 μL sample was pipetted, diluted with 980 μL methanol, vortexed, centrifuged at 14000×g for 2.0 min, filtered through a 0.2 μM PTFE filter and analyzed by HPLC for conversion and de. The reaction was stopped after 8 h (99.7% conversion).

[0199] The reaction mixture was acidified to pH 1.3 (6N HCl, 3.6 mL). A diatomaceous earth pad was prepared and the reaction mixture was filtered through the diatomaceous earth. After filtration, the reactor and diatomaceous earth pad were washed with 30 mL of water and the solution was poured out with the filtrate. The reaction mixture was extracted with 130 mL of 2-methyltetrahydrofuran and the 2-methyltetrahydrofuran solution was discarded. The pH of the aqueous layer was raised to 10.5 with 10N sodium hydroxide (4.4 mL). The aqueous layer was extracted with 150 mL of n-butanol (152 mL) (the volume of the organic layer was 190 mL and the volume of the aqueous layer was 105 mL) and the layers were separated. The aqueous layer was extracted again with 100 mL of n-butanol (105 mL) and the aqueous layer was separated from the organic layer (the volume of the aqueous layer was 75 mL and the volume of the organic layer was 125 mL).

[0200] The organic layer was poured out, and the solvent was concentrated to 40g of viscous liquid. The residue was solidified for 1 hour at 4°C. The residue was suspended in 240mL MTBE and stirred vigorously. The precipitate was filtered and the filtrate was discarded. The precipitate (6.6g) and 50mL water were stirred, and the pH was increased to 8.5. The desired compound was precipitated, filtered and washed with MTBE. The filtrate volume was reduced (15mL), the pH was increased to 9.0, and the precipitated compound was filtered and washed with MTBE. The two products were mixed and dried in a vacuum oven at 35°C overnight. 3.94g of the desired compound 13 in the form of a gray solid was isolated, with a yield of 78.8%, AP 99.3, de>99.9%, and a titer of 97%.

[0201] 1H NMR(400MHz,DMSO-d6)δ9.33(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 FWf + :336.24. Embodiment 11 Alternatively, compound 13 was synthesized

[0203] Hydrochloric acid (12N, 22.6m, 3.35eq.) was added to a 500mL reactor containing 139mL of water at 25°C over 90 minutes. Isopropylamine (4.05eq, 25.88mL) was then added until the pH reached 10.5. Compound 12 (25g) and pyridoxal-5'-phosphate (0.01g / g, 0.25g) were then loaded, followed by transaminase CDX-50 (0.02g / g, 0.50g). The reaction mixture was heated to 50°C and stirred for 24 hours. Hydrochloric acid (12N, 0.6eq, 4ml) and 1g of diatomaceous earth (4wt%) were loaded. The reaction mixture was heated to 80°C for 2 hours and then cooled to 20°C before filtering and washing. The solid residue was discarded and the mixture was heated to 50°C. Potassium hydroxide solution (10wt%, 65mL, 1.55eq) was then added until the pH reached 9.5. The mixture was then cooled to 20° C. over 3 hours. Compound 13 was isolated by filtration, washed with water and dried. Example 12 Synthesis of compound (I)

[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 minutes, a liquid of compound 13 (0.021 g, 0.062 mmol) in 0.5 ml CH3CN and DMF (0.1 ml) was added. The resulting solution was stirred at room temperature for 2 hours and then purified by reverse phase chromatography to yield compound (I) as its trifluoroacetic acid salt.

[0205] 1 H 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. Embodiment 13 Preparation of compound 21

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

[0208] Crystals of magnesium (8.73 g, 359 mmol, 1.35 eq.) and I2 were added to a 1L three-necked flask. Anhydrous THF (100 mL) was loaded into a reaction flask under N2. The reaction temperature was monitored by a J-chemical thermometer. 3-Bromo-1,1-dimethoxy-propane (65.8 g, 356 mmol, 1.35 eq.) was diluted with THF (150 mL) and loaded into an addition funnel. 3-Bromo-1,1-dimethoxy-propane solution (20 mL) was added to the flask at 20 ° C, and the entire light brown suspension was stirred vigorously to initiate the reaction using the friction between the stirring rod and the magnesium sheet. After 30 minutes, the light brown color disappeared, and the solution temperature rose to 45 to 50 ° C. The reactant temperature was then maintained between 55 and 62 ° C, while 3-Bromo-1,1-dimethoxy-propane solution was slowly added from the addition funnel. After 1.5 hours, the addition was completed, and the entire solution was kept at 60° C. for another two 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 eq.) were loaded into an addition funnel in THF (150 mL). The matrix solution was added to the Grignard solution within 20 minutes to generate a reddish yellow solution, and the internal temperature was kept below a maximum of 35° C. After 15 minutes, HPLC and TLC both showed that the reaction was complete. Water (20 mL) was slowly added to the crude mixture to precipitate all brown colloidal solids. All crude products were filtered through a diatomaceous earth pad and washed twice with a total amount of 100 mL of THF. The crude solution was dried over Na2SO4. The crude product was concentrated under vacuum at 30-35° C. to obtain compound 16 as a yellow oil.

[0209] Optional: The crude product was favored for the next reaction without purification. To obtain spectra: The crude product was then purified on an ISCO purification system using Hexane / EA 1:0 to 10:1 to give the desired product.

[0210] 1 H NMR (500MHz, CDCl3) δ8.60(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

[0212] In a 250 mL round bottom flask, compound 16 (7.61 g, 38.5 mmol, 1 eq.) was diluted with water (20 mL) and THF (80 mL) at 20 °C. Trifluoroacetic acid (8.5 mL, 110 mmol, 2.7 eq.) was added to the solution at room temperature. The solution was quickly warmed to 50 °C. After 4 hours, the solution turned dark brown, and HPLC and TLC showed that the reaction was complete. Additional water (60 mL) was added to the flask. Sodium bicarbonate (9.6 g, 114 mmol, 2.8 eq.) was slowly added to the crude product at 25 °C to neutralize the medium to pH 7. The crude material was extracted three times with EtOAc (100 mL), and the combined organic crude product was washed once with brine (50 mL). The crude product was dried over Na2SO4. The crude product was filtered and concentrated to a dark oil that was used directly in the next step. CH2Cl2 (75 mL) was added to the crude product at 20 °C. To the solution containing compound 17 was added ethyl-2-(triphenylphosphinoalkylidene) propionate (14.2 g, 38.2 mmol, 0.93 eq.) in one portion. The reaction was kept at 20 °C for 8 hours. The crude product was concentrated to dryness and diluted with 1:1 hexanes:EtOA, the solid precipitate was filtered off, and washed twice with MTBE (20 mL). The combined crude product was concentrated to a black oil and purified on an ISCO purification system (200 g silica gel) using hexanes / EA 1:0 to 5:1 to give the desired compound 18-Et (8.21 g, 70.3%) as a yellow oil.

[0213] 1 H NMR(500MHz, CDCl3)δ8.59(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

[0215] To compound 18-Et (9.55 g, 33.9 mmol, 1.0 eq.) was added p-toluenesulfonic acid (2.35 g, 13.5 mmol, 0.40 eq.), trimethyl orthoformate (24 mL, 220 mmol, 6.4 eq.) and methanol (95 mL) at 20°C. The solution was refluxed for 60 hours. The crude product was cooled to 0°C, and sodium hydroxide (1.7 mL, 17 mmol, 0.5 eq.) was added to neutralize the medium to pH 7. The crude product was evaporated to a viscous oil and diluted with MTBE (200 mL). The crude product was washed once with water and brine. The crude product was dried over Na2SO4 and filtered. The crude product was filtered and swept over ISCO (80 g silica) using hexanes / EA 1:0 to 4:1 to give compound 19-Et (8.1 g, 73% yield) and ethyl (E)-6-(4-chloro-2-pyridinyl)-2-methyl-6-oxo-hexan-2-oate (1.2 g, 13%).

[0216] 1 H NMR (500 MHz, CDCl3) δ 8.50 (d, J = 4.0 Hz, 1H), 7.63 (d, J = 0.8 Hz, 1H), 7.17 (dd, J = 0.8 and 4.0 Hz 1H), 6.45 (dt, J = 8.0 and 0.4 Hz, 1H), 4.06 (q, J = 7.8 Hz, 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 = 7.8 Hz, 3H).

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

[0218] The pale yellow oil compound 19-Et (1.14 g, 3.48 mmol, 1.0 eq.) was diluted in ethanol (10 mL) at 20 ° C. Sodium hydroxide (2 mol / L) was added to the above solution in water (2 mL, 4 mmol, 1.1 eq.) at room temperature. The solution was heated to 60 ° C for 12 hours. HCl (1 mol / L) (4 mL) was added to the crude product, and the entire crude product was evaporated to generate a white paste. Saturated aqueous NH4Cl (20 mL) was then added to the crude product and extracted twice with 2-methyl THF (10 mL). The crude product was dried on Na2SO4 and filtered, concentrated to generate a pink crude product. The crude product was filtered and swept over ISCO (8 g silica) using hexane / EA 1: 0 to 2: 1 to generate compound 20 (1.05 g, 100%) as white crystals.

[0219] 1 H NMR (500MHz, CDCl3) δ8.61(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

[0221] Compound 20 (632 mg, 2.11 mmol, 1.0 eq.) and a pressurized reactor were placed in a glove box. Subsequently, catalyst diacetoxy [(R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] ruthenium (II) (95 mg, 0.109 mmol, 5.2 mmol%) and methanol (5 mL) were added to the glove box, and a vial was placed in the reactor and sealed. The reactor was placed on a hydrogenation stand and swept with hydrogen several times. The reaction was carried out at room temperature under 150 psi H2. After 12 hours, the reactants were removed from the reactor, and the solution turned reddish black. TLC and LCMS showed that the reaction was complete. After concentrating the crude product, the crude product was placed on an ISCO (silica, 8 g) with hexane / EA 1:0 to 1:1 to generate the desired product compound 21 (0.63 g, 99%) as a brown solid. Chiral HPLC determined 92% ee of the desired product.

[0222] 1H NMR (500MHz, CDCl3) δ8.59(d,J=5.2Hz,1H),7.69(d,J=1.8Hz,1H),7.25(dd,J=5.2and 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. Embodiment 14

[0224] Compound 37a (2.8 g) and methyl 2-(triphenyl-15-phosphinyl)propanoate (11 g) were dissolved in DCM (100 mL) and stirred at room temperature for 4 hours. After complete conversion, the solvent was removed under low pressure, and the obtained residue was purified by silica gel chromatography to give 6.9 g of the desired compound 38a.

[0225] 1 H NMR (CDCl3, 400MHz): δ6.69ppm, (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 the conversion was completed, 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 obtained crude compound 39a (11 g) was dissolved in ACN (220 mL), water (110 mL) and methanesulfonic acid (100.5 g, 34.967 eq). The reactants were warmed to 65 ° C until the conversion was completed. After cooling to room temperature, the pH was adjusted to 5-7 with concentrated NaOH solution. The organic layer was extracted with EtOAc (500 mL×2), dried over Na2SO4, and filtered to provide 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 warmed to 45-50 ° C and stirred overnight. After cooling to room temperature, the 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 reduced under low pressure. The obtained residue was purified by column chromatography on silica gel using petroleum ether / EtOAc (50:1 to 3:1) to provide compound 19a (10.4 g). 1 H NMR (CDCl3, 400MHz): δ8.56ppm (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). Embodiment 15

[0228] Under nitrogen, paraformaldehyde (1.44g, 1.5eq) is loaded into the reactor, followed by MeOH (25ml, 5vol), followed by 2-oxocyclopentane-1-carboxylic acid ethyl ester (5g, 1eq) and finally diethylamine (7.02ml, 2.1eq). The reaction mixture is stirred at 20-25°C for at least 3.5 hours. After completing the conversion, sodium hydroxide (6M, 26.6ml, 5eq) is loaded, and stirred at 20-25°C for at least 2 hours until the conversion is completed. MeOH is distilled out under vacuum. Subsequently, MTBE (25ml, 5vol) is loaded into the reaction mixture, and stirred for 10 minutes. Each layer can be determined, then discharged and the upper organic layer is discarded. The 6M HCl aqueous solution is added to the lower aqueous layer to pH 2. EtOAc (25ml, 5vol) is loaded, and the reaction mixture is stirred for at least 10 minutes. After phase separation, it is washed with water. The organic layer was concentrated to dryness to generate compound 41a, which was directly dissolved in iPrOH (50 mL, 10 vol.) and H2SO4 (1 eq.) at room temperature and stirred for 72 hours. The pH of the reaction mixture was adjusted to 7.3 with 20% aq. K2HPO4. After phase separation, the iPrOH layer was diluted with 20% aq. K2HPO4 (10 vol.) and washed with toluene. Finally, the pH of the aqueous layer was adjusted to 4.7 with 2M H2SO4. The desired compound 42a was extracted with toluene and isolated after removing the solvent under low pressure.

[0229] 1 H NMR (400MHz, CDCl3): δ6.33ppm(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 eq) was loaded into a nitrogen-swept 3-necked flask at 20-25° C., followed by compound 42a (1 g, 1 eq) at 20-25° C. and THF (10 mL, 10 vol) at 20-25° C. After cooling to -30° C., 1M LiHMDS in THF (12.49 mL, 2.5 eq) was added dropwise over 30 minutes, keeping the temperature ≤ -25° C., and stirred at -20° C. for at least 1 hour. After complete conversion, acetic acid (0.9 mL, 3 eq) was added dropwise, keeping T < -10° C., and the reaction mixture was allowed to warm to RT. EtOAc (50 mL, 50 vol) was added, and the organic layer was washed with water and brine. After concentration under low pressure, aqueous solution (15 mL, 15 vol) and sulfuric acid (6.7 mL, 25 eq) were slowly added. The reaction mixture was then heated to 65° C. and maintained at 65° C. for at least 17 hours. The reaction mixture was 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 (5g) was dissolved in TMOF (4eq), H2SO4 (1.1eq), MeOH (4vol), and stirred at 50°C overnight. After conversion was completed, NaOH (6.4M, 8eq) was added and stirred for 2 hours. After MeOH was distilled under low pressure, DCM was added to the reaction mixture, and 30% aq citric acid was used to adjust the pH to 5. After DCM was extracted, the organic layer was washed with water and concentrated to dryness. The obtained residue was dissolved in MeCN (2.5vol) and heated to 45°C. The reactant was slowly cooled to 0°C and stirred for 1 hour. Finally, water (10vol) was added in 2.5vol portions. After filtering and washing the filter cake, compound 19b was obtained as a solid with a yield of 88%.

[0233] 1 H NMR (500MHz, CDCl3): δ8.6ppm(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] [RuCl(isopropyltoluene)((R)-H8-binap)]Cl (0.0003 mmol) was added in a DCM stock solution (100 μL) to a 5 mL vial equipped with a magnetic stirrer. Subsequently, compound 19b (0.075 mmol) was added as a stock solution of MeOH / DCM 3 / 1 (1 mL), followed by TEA (0.375 mmol). The vial was capped and transferred to a B48 parallel reactor. The reaction was carried out at 25 °C under H2 atmosphere (40 bar) overnight (ca. 16 hours). The reactants were analyzed by HPLC to provide the desired compound 21. Example 16 Crystallization of compound 21

[0235] Crude compound 21 (1.0 g) was suspended in 25-30 mL of heptane and stirred to 40° C. to a clear solution. After cooling to 35° C., the reaction mixture was seeded and the mixture was stirred for 2-4 hours and then cooled to -5° C. over 8-10 hours. After 6-10 hours at -5° C., the filter cake was filtered, washed, and dried in an oven at 30° C. mp: 64° C. Embodiment 17 Enzymatic method for the synthesis of compound 27 a) Synthesis of compounds 24a and 25a

[0236] Potassium tert-butoxide (1 mol / L) was added to a mixture slurry of 2-methylcyclopentanone, compound 22 (93.30 g, 931.7 mmol, 98 mass %), compound 2a (158.02 g, 902.55 mmol, 98 mass %) in THF (1200 g, 1330 mmol, 1 mol / L) at -30°C. The resulting yellow slurry was stirred between -24 and 30°C for 1 hour. Sulfuric acid (13.14 mol / L) in water (92 g, 660.2 mmol, 13.14 mol / L) and water (800 g, 44407.9 mmol, 100 mass %) were loaded into another 4 L reactor and pre-cooled to 0°C. The yellow slurry containing compound 23a-Cl was poured into the cooled acid solution and a slurry was produced. THF was distilled off at 15°C under vacuum (115 mbar) using a cannula set to 45°C. 500mL water is added to the slurry (~1L). The precipitated solid is collected and the aqueous solution is discarded. The collected solid is reloaded into the reactor and 320mL MSA and 1L water are loaded simultaneously. The slurry is heated to 65°C and all solids are dissolved after 60 minutes. The dark solution is kept at 65°C for 3 hours, then cooled to room temperature and then cooled to 0°C. A slurry is formed and filtered. Compound 24a is collected and dried at room temperature to obtain a total amount of 139.8g of gray-brown solid. The filtrate is reloaded into the reactor and the pH is adjusted to 5.1 with 28wt% NH4OH. Solids are formed during the pH adjustment and filtered at room temperature. Additional 41g of gray solid compound 24a is obtained.

[0237] 1 H NMR (400MHz, DMSO-d6): δ12.07(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 loaded into a 2 L reactor, followed by 3.4 L MeOH, 380 mL TMOF, 210 mL TMSCl. The mixture was heated to 49 ° C, and the cannula was set to 57 ° C. After another 4 hours at 50 ° C, the dark solution was cooled to 10 ° C, and then NaHCO (2.6 L saturated) solution was loaded into a 20 L reactor, with a total volume of 6.5 L.

[0240] Most of the MeOH solvent was distilled off using a cannula under vacuum at 35° C. to a volume of 3.3 L. 2 L MTBE was then added. The organic layer was separated from the aqueous layer and concentrated to provide 207 g of Compound 25a as a colored liquid.

[0241] 1 H NMR (400MHz, DMSO-d6): δ8.60(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. Kilogram batch

[0243] Potassium tert-butoxide (258 kg, 0.73X, 1.1 eq.) was added portionwise to a mixture of 2-methylcyclopentanone, compound 22 (235 kg, 0.66X, 1.16 eq.) and compound 2a (354 kg, 1.0X) in 2-Me-THF (2103 L, 5.1X, 5.9 V) at 0° C. over 5 hours under N2. After 2 hours, the reaction mixture was quenched with water (2839 kg, 8X, 8V; pre-cooled to 3-8° C.) at 0° C. over 4 hours. 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 adding 5% H2SO4 aqueous solution (1970 kg, 5.6X, 0.49 eq.) at 0° C. within 5.5 hours, and then further adjusted to 4.0-6.0 (4.92) by adding 0.5% H2SO4 solution (611 kg, 1.7X, 0.02 eq.) at 0° C. within 2 hours. The mixture was stirred at 0° C. for 30 minutes, then filtered by centrifugation and washed with water (1495 kg, 4.2X, 4.2V) to obtain 565 kg of compound 23 as a wet solid.

[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), H2O (90.00 g, 90 ml) and ACN (29.25 kg, 37 L). The reaction mixture was heated to 68°C and stirred for 5 hours. The reaction mixture was cooled to 20°C, then 25% ammonia solution (5.50 kg) was added portionwise to the reaction mixture and stirred for 1 hour. 2.5% ammonia solution (1.50 g) was added portionwise to the reaction mixture over 30 minutes to pH 4.8. The reaction mixture was heated to 43°C for 8 hours, then filtered to obtain 40 kg of compound 24a.

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

[0246] During the reaction, a jacketed 250 mL reactor with a water circulator that maintains the reaction temperature at 35°C is used. The calibrated pH state is used to maintain the pH at 7.0. 5.0 g of compound 25a, 4.0 ml (2%) DMSO and 180 ml sodium phosphate buffer (0.1 M, pH 7.0) are loaded into the 250 mL reactor. 5 N sodium hydroxide is continuously supplied to maintain the reaction at pH 7.0. Lipase MH Amano 10SD 1.0 g is dissolved in 10 mL of the same buffer and added to the reactor. Another 6.0 mL of buffer is used to wash the enzyme container and load it into the same reactor. The sample (80 μL) is aspirated with a dropper, diluted with 1.920 mL of methanol, vortexed, centrifuged for 2 minutes, filtered and analyzed by non-chiral and chiral HPLC. The reaction stops after 23 hours, and the pH is raised to 8.2 by using 10 N sodium hydroxide. The reactants are extracted with 200 mL of ethyl acetate. All dimethoxymethyl esters are extracted into the organic layer. The organic layer was extracted again with sodium potassium buffer (2×50 mL) at pH 8.5 to remove the 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 was removed under vacuum, and the residue was dried in a vacuum oven overnight. 1.88 g of compound 26a was isolated as a brownish liquid with a yield of 37.6% and ee of 98.7%.

[0247] 1 H NMR (400MHz, DMSO-d6) δ8.61(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: [C 15 H 22 ClNO4-OCH3] + , 284.2 / 286.1. c) Hydrolysis of compound 26a to generate compound 27a:

[0249] Compound 26a (1.0 g), methanol (20 mL), water (5.0 mL) and 500 μL (0.2 g) of 10 N sodium hydroxide were loaded into a reaction flask. The reaction was stirred at room temperature for 5 hours. A sample (30 μL) was removed, diluted with methanol (970 μL), vortexed, filtered and analyzed by HPLC. Most of the reaction (~98% conversion) stopped in <2 hours, and no racemization was observed during the hydrolysis. The reaction mixture was concentrated to an oil and diluted with 20 mL of water (pH 12.7). The reaction mixture was extracted with MTBE (2×50 mL) and the MTBE was discarded. The aqueous layer was cooled and acidified to pH 3.8 with 850 μL of 6N HCl. The aqueous layer was extracted with MTBE (2×50 mL), and the MTBE solution was washed with brine (25 mL) and water (2×25 mL). The MTBE solution was dried over anhydrous sodium sulfate, filtered, the solvent was removed, and the residue was dried in a vacuum oven overnight. 940 mg of compound 27a were isolated as a viscous yellow liquid in 98.3% yield, AP 97 and ee 98%.

[0250] 1 H NMR (400MHz, DMSO-d6) δ11.96(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 H 20 ClNO4] - , 300.1. Optional enzymatic analysis

[0252] At 20-25°C, 13.5 L of Tolis buffer 0.1 M and 50 mM Ca(OAc)2 were mixed with 2.5 w% enzyme (Almac hydrolase L90 enzyme, also known as AL-L90, commercially available from ALMAC Group Ltd., Craigavon, Northern Ireland, UK). The pH of the mixture was adjusted to 7.2-7.8. The mixture was heated to 38-42°C and the pH was maintained at 7.2-7.8. A portion of 1.5 kg of compound 25a in 1.5 L DMSO was added to the mixture at 38-42°C. After 22 hours, the conversion was 49%. The reaction mixture was cooled to 5°C and the mixture was maintained at 0-10°C for 16 hours. Development: 9 L CAN was added to the reaction mixture at 0-10°C. Subsequently, the pH was adjusted to 10.0 by 20% K2CO3 and 0.25× diatomaceous earth was added to the mixture. After stirring for 20-30 minutes, the reaction mixture was filtered and the filter cake was washed with 3L MTBE and 3L water. The filtrate was collected and 15L MTBE was added to the mixture for phase separation. The organic layer was washed twice with 15L water. The aqueous phase was combined and pH=5.5 was adjusted by 20% citric acid. 15L MTBE was added to the mixture for phase separation. The aqueous layer was then adjusted by 20% citric acid to pH=5.5, and 15L MTBE was used again for phase separation. The organic phase was combined and washed with 15L process water. The organic phase was collected and filtered to remove the aqueous layer. The organic phase was then concentrated to 4.5L. Two azeotropic distillations were performed to 4.5L using 15L MTBE. 7.5L MTBE and 3L ACN were added to the residue to form a salt. Salt formation: The mixture was heated to 50-55°C, and DCHA (0.75eq) was added to the mixture at 50-55°C. After stirring at 50-55°C for 1 hour, the reaction mixture was cooled at 10°C / hour. 1.8% seed crystals were added to the mixture at 38.2°C. A white solid was slowly precipitated. After keeping at 38°C for 3 hours, the mixture was continued to be cooled at 10°C / hour. After keeping at 0-5°C for 10 hours, the mixture was filtered. The filter cake was washed with 1.5L pre-cooled ACN. After drying for 38 hours, 1.245kg of compound 27a-DCHA as a white solid was obtained, which had 99.3% purity, 99.4% ee and 43.61% isolated yield. Embodiment 18 Synthesis of compound 10 a). Synthesis of compound 29a

[0253] n-Butyl lithium (2.5 mol / L) in hexane (22 mL, 55.0 mmol, 2.50 mol / L) was slowly added to a solution of (triisopropylsilyl)acetylene (10.5 g, 57.6 mmol, 100 mass %) in THF (60 mL) at -10 °C. After addition, the mixture was warmed to 21 °C. A solution of compound 28a (10.0 g, 49.8 mmol, 100 mass %) in THF (35 mL) was then added at 21 °C. After 1 hour, HPLC analysis showed that 8% of the amide starting material remained. Lithium bis(trimethylsilyl)amide (1 M in THF, 8 mL) was then added. After 1 hour, HPLC analysis showed that 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 MgSO 4 , and concentrated to give 15.8 g of compound 29a as an orange oil in 98.5% yield.

[0254] 1 H NMR (500MHz, CDCl3): δ8.68 (1H,d,J=4.5Hz), 8.15 (1H,s), 7.50 (1H,d,J=4.5Hz), 1.25-1.05 (21H,m).

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

[0256] Trimethyl orthoformate (2.0 mL, 18 mmol) was added to a solution of compound 29a (3.00 g, 9.32 mmol) in MeOH (20 mL) at 21 °C, followed by the addition of chlorotrimethylsilane (3.0 mL, 24 mmol). The mixture was then heated at 60 °C. After 1 hour, HPLC analysis showed that the starting material was consumed. The mixture was cooled to 21 °C and added to a mixture of hexanes (200 mL) and NaHCO (15 g) / NaCO (5 g) aqueous solution in water (200 mL). The separated organic phase was then dried over MgSO and concentrated to yield 3.45 g of [3-(4-chloro-2-pyridinyl)-3,3-dimethoxy-prop-1-ynyl]-triisopropyl-silane (3.45 g, 9.38 mmol, 100% yield) as an orange oil. The crude intermediate was used for subsequent deprotection of TIPS without further purification.

[0257] 1M tetrabutylammonium fluoride in THF (12 mL, 12.0 mmol, 1.00 mol / L) was added to a solution of [3-(4-chloro-2-pyridyl)-3,3-dimethoxy-prop-1-ynyl]-triisopropyl-silane (3.20 g, 8.70 mmol, 100 mass %) in 2-MeTHF (10 mL) and TBME (10 mL) at 21°C. After 10 minutes, 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 in 130 mL of water). The separated organic phase was dried over MgSO4 and concentrated. The resulting residue was purified by column chromatography (20 to 60% EtOAc / heptane; Rf 0.39 in 50% EtOAc / heptane) to give 1.65 g of compound 30a as a light-colored solid in 90% yield.

[0258] 1 H NMR (500MHz, CDCl3): δ8.61 (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 10 H 10 ClNO2–OCH3] + , 180.25 / 182.06. c) Synthesis of compound 31a

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

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

[0262] 1 H NMR (500MHz, CDCl3): δ8.57(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 11 BrClNO2–OCH3–Br] + , 182.09 / 184.22. d) Synthesis of compound 32a

[0264] At 5-10 ° C, (S)-(-)-3-methoxy-2-methyl-3-oxopropylzinc bromide (0.5 mol / L) in THF (94 mL, 47 mmol, 0.50 M) was loaded into a solution of compound 31a (12.0 g, 41.0 mmol, 100 mass %) in THF (50 mL). The mixture was degassed by bubbling with N2 for 3 minutes. Then 1,1'-bis (di-tert-butylphosphino) ferrocene palladium dichloride (0.68 g, 1.03 mmol, 100 mass %) was added at 5-10 ° C. The resulting mixture was degassed by bubbling with N2 for 5 minutes. After keeping at 5 ° C for 15 minutes, the dark solution was warmed to 21 ° C. After 16 hours, TBME (150 mL) was added to the reaction mixture, followed by NH4Cl aqueous solution (25%, 200 g). The dark organic phase was dried over MgSO4 and concentrated. 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 mass, 90.9% yield) as an orange oil.

[0265] 1H NMR (500MHz, CDCl3): δ8.54(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] Preparation of catalyst: Diacetoxypalladium (440 mg, 1.960 mmol) and bis(1-adamantyl)-butyl-phosphine (708 mg, 1.975 mmol) were added to 12 mL of dioxane. The mixture was degassed by bubbling with N2 for 0.5 h. Compound 32a (6.2 g, 20 mmol, 100 mass %), compound 8 (4.0 g, 25 mmol), pivalic acid (1.1 g, 11 mmol, 100 mass %), potassium carbonate (8.1 g, 59 mmol) were mixed in 60 mL of dioxane, and the resulting mixture was degassed by bubbling with N2 for 0.5 h. The premixed and degassed catalyst was then transferred to the mixture including the substrate. The resulting mixture was degassed by bubbling with N2 for 0.5 h, then heated at 90 ° C, and maintained at this temperature for 3 hours, and then cooled to 21 ° C. The resulting mixture was filtered and the 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 7.6 g of compound 33a (7.6 g, 16 mmol, 98 mass, 86% yield) as a brown oil.

[0268] 1H NMR (500MHz, CDCl3): δ8.86 (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 solution of compound 33 (6.3 g, 14 mmol) in MeOH (50 mL) and a magnetic stirring bar were loaded into a 100 mL pressurized flask. The bottle was degassed by vacuum / refilling N2 6 times. Pd / carbon, 10 wt%; 50% wet; (1.35 g, 0.634 mmol, 5 mass %) was then added. The container was degassed with N2 and then with H2. The hydrogen pressure was set to 80 psi and the reaction temperature was 55°C. After maintaining at this pressure and temperature for 12 hours, the mixture was filtered and the line was washed 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 this 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 4.85 g of compound 10a (4.85 g, 12.2 mmol, 89.7% yield) as a foaming solid.

[0271] 1H NMR (500MHz, CDCl3): δ8.77(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. Embodiment 19 Synthesis of compound 14 Suzuki

[0273] Acetonitrile (500 mL, 5 L / kg), compound 43 (100 g, 1.0 eq), compound 44 (60 g, 1.05 eq), 1,1,3,3-tetramethylguanidine (93.2 g, 2.05 eq) and water (70.8 g, 10 eq) were loaded into a 2 L reactor. The reactor headspace was purged with a stream of nitrogen. Pd(dppf)Cl2.DCM (3.2 g, 1 mol%) was added to the reaction mixture, and the reactor was heated to 65°C over 1-7 hours and stirred at this temperature for 1-17 hours. After the reaction was completed, 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 reaction mixture. Then warm water (280mL, 2.8L / kg) was slowly added to the reaction mixture over 0.5-4 hours while the internal temperature was maintained at 60-65°C. After aging for 1-4 hours, warm water (350mL, 3.5L / kg) was slowly added to the reaction mixture over 2-6 hours while the internal temperature was maintained at 60-65°C. Then the reaction mixture was cooled to 10-15°C over 4-6 hours and further aged for 1-3 hours at 10-15°C. The slurry was then filtered and the filter cake was washed with a cooled MeCN / water solution (2:1 v / v ratio, 5L / kg). The compound 45 filter cake was dried overnight under vacuum at 45-50°C.

[0274] Typical results: 90-95% yield, 98a% purity, >95% assay. Remove PD

[0275] The crude compound 45 solid (100 g, 1.0 eq), 2-methyltetrahydrofuran (MeTHF, 1.5 L, 15 L / kg) and N-acetyl L-cysteine ​​aqueous solution (32 g in 1.5 L water) were loaded into a 5 L reactor. The resulting mixture was stirred at 20-30 ° C for 1-2 hours, and filtered through diatomaceous earth (30 g, 0.3 kg / kg). 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 reactor, and the N-acetyl L-cysteine ​​aqueous solution (32 g in 1.5 L water) was loaded again. 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 reactor, and aqueous sodium bicarbonate (70 g in 1 L water) was loaded. 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 reactor and aqueous sodium sulfate (100 g in 1 L of water) was loaded. 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 reactor and diluted with MeTHF (1 L, 10 L / kg), and the resulting solution was concentrated to 500-600 mL (5-6 L / kg) under vacuum. The solution was then diluted with MeTHF (1 L, 10 L / kg), and the resulting solution was concentrated to 500-600 mL (5-6 L / kg) under vacuum. The solution was then diluted with MeTHF (1 L, 10 L / kg), and the pharmaceutical water content (KF less than 0.1%).

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

[0277] Compound 45 MeTHF solution (100 g in 1.5 L MeTHF), MeTHF (1.5 L, 15 L / kg) and MeCN (1 L, 10 L / kg) were loaded into the reactor, and the resulting mixture was cooled to 5-10° C. TMSN3 (59.0 g, 1.2 eq) was slowly added to the reactor. tBuONO (53.0 g, 1.2 eq) was slowly added to the reactor, and the mixture was stirred at 5-15° C. for 4-8 hours. NaOH aqueous solution (100 g in 1 L water) was slowly added to the mixture, and then the mixture was warmed to 20-30° C., stirred for 20-30 minutes, and allowed to stand at 20-30° C. for 30-60 minutes. After phase separation, the upper layer was left in the reactor, and NaOH aqueous solution (100 g in 1 L water) was slowly added to the mixture, and then the mixture was stirred for 20-30 minutes and allowed to stand at 20-30° C. for 30-60 minutes. After phase separation, the upper layer is left in the reactor, and an aqueous NaOH solution (100 g in 1 L of water) is slowly added to the mixture, followed by stirring the mixture for 20-30 minutes, and standing at 20-30° C. for 30-60 minutes. After phase separation, the upper layer is left in the reactor, and an aqueous NaOH solution (100 g in 1 L of water) is slowly added to the mixture, followed by stirring the mixture for 20-30 minutes, and standing at 20-30° C. for 30-60 minutes. After phase separation, the upper layer is left in the reactor, and pharmaceutical residual azide (residual N3 <3 ppm). An aqueous sodium sulfate solution (100 g in 1 L of water) is then added to the mixture, followed by stirring the mixture for 20-30 minutes, and standing at 20-30° C. for 30-60 minutes. After phase separation, an aqueous sodium sulfate solution (100 g in 1 L of water) is added to the mixture, followed by stirring the mixture for 20-30 minutes and standing at 20-30° C. for 30-60 minutes. After phase separation, an aqueous sodium sulfate solution (100 g in 1 L of water) is added to the mixture, followed by stirring the mixture for 20-30 minutes and standing at 20-30° C. for 30-60 minutes. After phase separation, an aqueous sodium sulfate solution (100 g in 1 L of water) is added to the mixture, followed by stirring the mixture for 20-30 minutes and standing at 20-30° C. for 30-60 minutes. After phase separation, an aqueous sodium sulfate solution (100 g in 1 L of water) is added to the mixture, followed by stirring the mixture for 20-30 minutes and standing at 20-30° C. for 30-60 minutes. After phase separation, a pharmaceutical pH (pH <9) and the organic azide solution is cooled to 5-15° C. After bubbling with nitrogen for 20-40 minutes, triethylamine (95.0 g, 2.2 eq) was slowly charged into the reaction mixture while the temperature was maintained at 5-15° C. Triethylsilylacetylene (50.0 g, 1.2 eq) was slowly charged into the reaction mixture while the temperature was maintained at 5-15° C. and the reactor was purged with nitrogen until the oxygen content was less than 0.1%.Copper iodide (8.0 g, 10 mol%) was loaded into the reactor and then purged with nitrogen again until the oxygen content was less than 0.1%. The reaction mixture was stirred at 5-15 ° C for 8-16 hours. After the reaction was completed, 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 diatomaceous earth (30 g, 0.3 kg / kg), and the filter cake was rinsed with MeTHF (250 mL, 2.5 L / kg). Aqueous ammonia solution (100 g in 1 L water) was loaded into the reactor, then stirred at 20-30 ° C for 20-30 minutes, and allowed to stand at 20-30 ° C for 30-60 minutes. After phase separation, an aqueous sodium sulfate solution (100 g in 1 L of water) was loaded into the reactor, followed by stirring at 20-30 ° C for 20-30 minutes, and allowed to stand at 20-30 ° C for 30-60 minutes. After phase separation, the upper layer was filtered through diatomaceous earth (30 g, 0.3 kg / kg). After washing the diatomaceous earth filter cake with MeTHF (0.5 L, 5 L / kg), the mixture was concentrated to 500-700 mL (5-7 L / kg) at 45 ° C under low pressure. N-heptane (1 L, 10 L / kg) was added dropwise to the reactor, and the resulting mixture was concentrated to 500-700 mL (5-7 L / kg) at 45 ° C under low pressure. N-heptane (1 L, 10 L / kg) was added dropwise to the reactor, and the resulting mixture was concentrated to 500-700 mL (5-7 L / kg) at 45 ° C under low pressure. The reaction mixture was then heated to 55-60° C. and stirred at this temperature for 2-4 hours. After the reactor was cooled to 5-15° C. over 3-8 hours, the slurry was aged at 5-15° C. and filtered. The compound 46 filter cake was washed with n-heptane (1 L, 10 L / kg) and dried at 40-45° C. under reduced pressure for 6-12 hours.

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

[0279] Compound 46 (100 g, 1.0 eq), DMF (500 mL, 5 L / kg) and purified water (5 g, 1.0 eq) cooled to -15- (-5 ° C) were loaded into the reactor. 1,3-dichloro-5,5-dimethylhydantoin (DCDMH, 13.7 g, 0.75 eq) was loaded portionwise while maintaining the internal temperature at 5 ° C. The internal temperature was then adjusted to 0-10 ° C, and the mixture was stirred at this temperature for 5-12 hours. After the reaction was completed, water (70 mL, 0.7 L / kg) was added to the reaction mixture within 0.5-1.5 hours while maintaining the internal temperature at 15 ° C. Compound 47 seeds (0.1 g, 0.001 kg / kg) were added to the reaction mixture and then matured at 0-10 ° C for 1-2 hours. Water (530 mL, L / kg) was added at 0-10° C. over 3-8 hours, and the slurry was aged at 0-10° C. for 4-6 hours. The mixture was filtered, and the filter cake was washed with cold water (0.5 L at 0-10° C.). The crude compound 47 was dried at 40-50° C. under reduced pressure for 8-15 hours.

[0280] Typical results: 90-95% yield, 98a% purity, >95% assay. Recrystallization of compound 47

[0281] The crude compound 47 (100 g, 1.0 eq), DCM (0.5 L, 5 L / kg) were loaded into the reactor, and the resulting solution was stirred at 20-30 ° C for 0.5-2 hours. The resulting mixture was filtered through a filter cartridge equipped with charcoal and circulated for 3-8 hours, and then concentrated to 300-360 mL (3.0-3.6 L / kg) at low pressure. The mixture was then warmed to 35-45 ° C and refluxed for 20-40 minutes, then cooled to 0-10 ° C within 1-4 hours and further matured at 0-10 ° C for 0.5-2 hours. N-heptane (1.6 L, 16 L / kg) was then loaded into the reactor at 0-10 ° C within 1-3 hours, and the slurry was matured at 0-10 ° C for 1-3 hours. After filtering the slurry, the pure compound 47 cake was washed with n-heptane (500 mL, 5 L / kg) and dried at 40-50 °C under reduced pressure for 6-12 hours.

[0282] Typical results: 90-95% yield, >99.5a% purity, >95% assay. Demethyl

[0283] Pure compound 47 (100 g, 1.0 eq), aqueous HCl (35 w / w%, 320 g, 10 eq) were loaded into the reactor, and the reactor was heated to 40-50 ° C. within 1-3 hours, and stirred at this temperature for 10-18 hours. After the reaction was completed, an aqueous ammonia solution (50 g in 500 mL) was added dropwise to the reactor at 40-50 ° C. within 2-6 hours to achieve pH = 5-7. The reaction mixture was then cooled to 0-10 ° C. within 1-3 hours and matured at this temperature for 1-3 hours. After filtering the slurry, the crude compound 14 filter cake was washed with cold water (1 L at 0-10 ° C., 10 L / kg) and dried at 40-50 ° C. under low pressure for 12-24 hours.

[0284] Typical results: 90-95% yield, >99.5a% purity, >95% assay. Recrystallization of compound 14

[0285] The crude compound 14 (100 g, 1.0 eq), acetone (1.4 L, 14 L / kg) were loaded into the reactor, and the reactor was heated to 50-60° C. and stirred at this temperature for 1-3 hours. After inoculation of compound 14 (0.5 g, 0.005 kg / kg), n-heptane (1.7 L, 17 L / kg) was added at 50-60° C. over 4-8 hours, and the mixture was stirred at this temperature for 1-2 hours, then cooled to 5-15° C. over 2-4 hours. After filtration, the pure compound 14 filter cake was washed with cooled n-heptane (0.5 L, 5 L / kg) and dried at 80-90° C. under reduced pressure for 8-16 hours.

[0286] Typical results: 90-95% yield, >99.9a% purity, >95% assay.

[0287] This application also relates to the following implementations: Item 1. A method for preparing compound (I), It contains the following steps: 1) Compound 1 having the following structure With the following compound 2 in X is selected from Cl, Br and I; Y is selected from OR 9 NHOC 1-3 alkyl, Cl, Br and I; and R 9 Selected from C 1-3 Alkyl, C1-3 Hydroxyalkyl, substituted phenyl and substituted benzyl; react in a suitable solvent to generate the following compound 3a or 3b 2) In the presence of an acid, convert compound 3a or 3b into the following compound 4 3) Compound 4 is then reacted with trialkyl orthoformate in an alcohol solvent to generate compound 5 of the following formula: in R 1 It is C 1-6 alkyl; and R 3 'Selected from C 1-6 alkyl, optionally substituted phenyl and benzyl; 4) Its ester hydrolysis under alkaline conditions or when R 3 When ' is a substituted benzyl group, hydrogenation is performed to form the following compound 6 Where R 1 and X is as defined above; 5) The carboxylic acid portion of compound 6 is then activated and reacted with a chiral auxiliary to form compound 6a of the following formula: Where Xa is a chiral auxiliary and R 1 and X is as defined above; 6) Compound 6a is then reacted with a base in the presence of a chiral auxiliary and a methyl donor such as a halogenated alkyl to form compound 7 Where R 1 , X and Xa are as defined above; 7) Remove Xa to obtain the following compound 7 Where R 1 and X is as defined above; 8) Then, in the presence of a metal catalyst, compound 7 is reacted with compound 8 having the following structure: To generate compound 9 9) Reducing the nitro group in compound 9 to form the following compound 10 10) Cyclization of compound 10 using a suitable coupling agent to generate compound 11 11) In the presence of an acid, the keto functional group is unmasked to generate compound 12 12a) using an ammonia equivalent in the presence of a reducing agent or 12b) using a transaminase in the presence of an ammonia source, a different recycling system and a cofactor to reduce compound 12 to generate the amino stereocenter in compound 13 13) which is then coupled with compound 14 of the following structure To generate compound (I) Item 2. A method as in Item 1, wherein in step 2), the acid is selected from sulfuric acid, methanesulfonic acid (MSA), benzenesulfonic acid, nitric acid, hydrochloric acid, trichloroacetic acid and perchloric acid. Item 3. The method of Item 1, wherein in step 5), the chiral auxiliary agent is selected from and in R 6 Selected from C 1-3 Alkyl, phenyl, benzyl; R 7 is selected from H and phenyl; and R 8 Selected from C 1-3 Alkyl, phenyl and benzyl. Item 4. A method as in Item 3, wherein the chiral auxiliary is selected from and Item 5. A method as in Item 1, wherein in step 6), the methyl donor is a halogenated alkyl group. Item 6. A method as in Item 1, wherein in step 8), the metal catalyst comprises palladium and a phosphine ligand. Item 7. A method as in Item 6, wherein the phosphine ligand is XPhos or SPhos. Item 8. The method of Item 1, wherein in step 10), the coupling agent is chloro-N,N,N',N'-tetramethylformamidine hexafluorophosphate (TCFH). Item 9. The method of Item 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. Item 10. A method as in Item 1, wherein in step 12b), the transamination reaction is carried out in the presence of (4-formyl-5-hydroxy-6-methylpyridin-3-yl)methyl phosphate (PLP). Item 11. A method as in Item 1, wherein in step 12b), the ammonia source is selected from isopropylamine, alanine, 3-aminobutyric acid and methylbenzylamine. Item 12. A method as in Item 11, wherein the ammonia source is isopropylamine. Item 13. A method for making a compound of formula (II) having a structure designated as Compound 21: The method comprises the following steps: a) making the following compound 15 With (CH3O)2CH(CH 2)2 Br reaction to generate the following compound 16 b) In the presence of an acid, converting compound 16 into compound 17 c) reacting compound 17 with a triphenylphosphonium alkane or phosphonate derivative in the presence of a suitable solvent to produce compound 18 Where R 3 'It's C 1-6 alkyl; d) reacting compound 18 with trimethyl orthoformate in the presence of an acid to produce compound 19 of the following formula: e) converting compound 19 into compound 20 having the following structure f) hydrogenating compound 20 in the presence of a ruthenium catalyst to generate a compound having the following structure: (II) Compounds Item 14. A method as described in Item 13, in step b), wherein the acid is TFA. Item 15. A method as in Item 13, wherein in step f), the acid is selected from hydrochloric acid, sulfuric acid, methanesulfonic acid and p-toluenesulfonic acid. Item 16. A method as in Item 13, wherein in step f), the ruthenium catalyst is selected from dichloro[(R)-(+)-2,2',6,6'-tetramethoxy-4,4'-bis(diphenylphosphino)-3,3'-bipyridine][(1R,2R)-(+)-1,2-diphenylethylenediamine]ruthenium(II), dichloro[(R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl][(1R,2R-)-(+)-1,2-diphenylethylenediamine]ruthenium(II), dichloro[(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]ruthenium(II) and diacetoxy[(r)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]ruthenium(II). Item 17. A method for preparing a compound of formula (III), in R 1 It is methyl; R 2 It is methyl; R 4 It is NO2 and R 5 It is CHF2; The method comprises the following steps: 1) Compound 28 of the following formula wherein X is selected from F, Cl, Br and I, Reaction with (triisopropylsilyl)acetylene to produce the following compound 29 2) Convert compound 29 into the following compound 30 3) Compound 30 is then reacted with bis(cyclopentadienyl)zirconium dichloride to produce a compound of the formula 4) Compound 31 is then reacted with (S)-(-)-3-methoxy-2-methyl-3-oxopropylzinc bromide to form compound 32 of the following formula: Where R 1 and X is as defined above; 5) Then, in the presence of a metal catalyst, compound 32 is reacted with the following compound 8: To generate compound 33 Item 18. A method according to Item 1, further comprising preparing compound 14: The method comprises the following steps: 1) Compound 43 of the following structure: With the following structure compound 44 The reaction is carried out under appropriate Suzuki coupling conditions in a suitable solvent to generate compound 45 with the following structure: 2) Compound 45 is converted into an azide, and the azide is reacted with trimethylsilyl acetylene and a metal catalyst in a suitable solvent to generate compound 46 having the following structure: 3) reacting compound 46 with 1,3-dichloro-5,5-dimethylhydantoin in a suitable solvent to generate compound 47 having the following structure: and 4) Compound 47 is demethylated in hydrochloric acid to give compound 14.

Claims

1. A method for preparing compound (I), It contains the following steps: 1) Compound 1 having the following structure With the following compound 2 in X is selected from Cl, Br and I; Y is selected from OR 9 NHOC 1-3 alkyl, Cl, Br and I; and R 9 Selected from C 1-3 Alkyl, C 1-3 Hydroxyalkyl, substituted phenyl and substituted benzyl; react in a suitable solvent to generate the following compound 3a or 3b 2) In the presence of an acid, convert compound 3a or 3b into the following compound 4 3) Compound 4 is then reacted with trialkyl orthoformate in an alcohol solvent to generate compound 5 of the following formula: in R 1 It is C 1-6 alkyl; and R 3 'Selected from C 1-6 alkyl, optionally substituted phenyl and benzyl; 4) Its ester hydrolysis under alkaline conditions or when R 3 When ' is a substituted benzyl group, hydrogenation is performed to form the following compound 6 Where R 1 and X is as defined above; 5) The carboxylic acid portion of compound 6 is then activated and reacted with a chiral auxiliary to form compound 6a of the following formula: Where Xa is a chiral auxiliary and R 1 and X is as defined above; 6) Compound 6a is then reacted with a base in the presence of a chiral auxiliary and a methyl donor such as a halogenated alkyl to form compound 7 Where R 1 , X and Xa are as defined above; 7) Remove Xa to obtain the following compound 7 Where R 1 and X is as defined above; 8) Then, in the presence of a metal catalyst, compound 7 is reacted with compound 8 having the following structure: To generate compound 9 9) Reducing the nitro group in compound 9 to form the following compound 10 10) Cyclization of compound 10 using a suitable coupling agent to generate compound 11 11) In the presence of an acid, the keto functional group is unmasked to generate compound 12 12a) using an ammonia equivalent in the presence of a reducing agent or 12b) using a transaminase in the presence of an amine source, a different recycling system and a cofactor to reduce compound 12 to generate the amine stereocenter in compound 13 13) which is then coupled with compound 14 of the following structure To generate compound (I) 2. The method of claim 1, wherein in step 2), the acid is selected from sulfuric acid, methanesulfonic acid (MSA), benzenesulfonic acid, nitric acid, hydrochloric acid, trichloroacetic acid and perchloric acid.

3. The method of claim 1, wherein in step 5), the chiral auxiliary is selected from in R 6 Selected from C 1-3 Alkyl, phenyl, benzyl; R 7 is selected from H and phenyl; and R 8 Selected from C 1-3 Alkyl, phenyl and benzyl.

4. The method of claim 3, wherein the chiral auxiliary is selected from 5. The method of claim 1, in step 6), wherein the methyl donor is a halogenated alkyl group.

6. The method of claim 1, wherein in step 8), the metal catalyst comprises palladium and a phosphine ligand.

7. The method of claim 6, wherein the phosphine ligand is XPhos or SPhos.

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

9. The method of claim 1, in step 12b), wherein the amine transaminase is selected from the group consisting of ATA-113, ATA-200, ATA-237, ATA-251, ATA-254, ATA-256 and ATA-260.

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

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