Synthesis of ras inhibitors

Abstract

The present invention relates to Ras inhibitors, intermediates in the synthesis thereof and processes for preparing said Ras inhibitors and said intermediates.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480063168.9 (22) Application Date 2024.10.02 (30) Priority Data 63 / 542112 2023.10.03 US 63 / 560883 2024.03.04 US (85) PCT International Application Entering National Phase Date 2026.03.31 (86) PCT International Application Application Data PCT / US2024 / 049574 2024.10.02 (87) PCT International Application Publication Data WO2025 / 076071 EN 2025.04.10 (71) Applicant: Ruixin Pharmaceutical Company, Address: California, USA (72) Inventors: Li Yi B. Mendoza S. Naganatan Wang Zhijian Yi Yi S.G. Ballmer J. Davidson Huang Xiaojun (74) Patent Agency: China Patent Agency (Hong Kong) Limited 72001 Patent Attorneys: Wang Yingyu Peng Chang (51) Int.Cl. C07D 203 / 10 (2006.01) C07D 401 / 04 (2006.01) C07D 401 / 14 (2006.01) C07D 413 / 06 (2006.01) C07D 487 / 10 (2006.01) C07D 519 / 00 (2006.01) (54) Title of Invention Synthesis of Ras Inhibitors (57) Abstract This invention relates to Ras inhibitors, intermediates in their synthesis, and methods for preparing said Ras inhibitors and said intermediates. Claims 11 pages, Description 123 pages, Claims amended according to Article 19 of the Treaty 14 pages, CN 122270438 A 2026.06.23 CN 1 22 27 04 38 A 1. A method for separating enantiomers of a compound of formula I, the method comprising the steps of: a) contacting the compound of formula I with a chiral acid to form a diastereomeric salt of the compound of formula I; and b) separating each diastereomeric salt of each diastereomeric isomer, thereby separating the enantiomers of the compound of formula I, wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 2. The method of claim 1, wherein the chiral acid comprises a chiral carboxylic acid. 3. The method of claim 1 or 2, wherein the separation step (b) comprises recrystallization. 4. The method according to any one of claims 1 to 3, wherein the compound of formula I is further described by formula Ia: . 5. A barium salt of a compound A: . 6.The barium salt of claim 5, wherein the barium salt is a 2:1 (compound A:barium) salt. 7. A method for preparing the barium salt of claim 5 or 6, the method comprising the steps of: a) condensing a compound of formula IIa with compound C to form a compound of formula IIb; b) cyclizing the compound of formula IIb with a compound of formula IIc to form a compound of formula IId; c) hydrolyzing the compound of formula IId to form a compound of formula IIe; d) methylating the compound of formula IIe to form a compound of formula IIf; and e) hydrolyzing the compound of formula IIf to form a barium salt of compound A: wherein R2 and R3 are each independently independently a optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-C10 aryl, and LG is a detached group selected from the group consisting of: halogen, trifluoromethanesulfonate, methanesulfonate, and toluenesulfonate. 8. The method of claim 7, wherein the methylation step (d) comprises contacting the compound of formula IIe with MeB(OH)2. 9. The method of claim 7 or 8, wherein the hydrolysis step (e) comprises contacting the compound of formula IIf with barium hydroxide. 10. A method for preparing a compound of formula III or a salt thereof: The method comprises the following steps: Claims 2 / 11 Page 3 CN 122270438 A a) coupling a compound of formula IIId with a compound of formula I to form a compound of formula IIIe; and b) deprotecting the compound of formula IIIe to form the compound of formula III: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; each R4 is independently an optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 11. The method of claim 10, wherein the compound of formula IIId is prepared by a method comprising the steps of: A1) combining the compound of formula IIIa with the compound of formula IIIb to form the compound of formula IIIc; B1) reducing the compound of formula IIIc to form the compound of formula IId; wherein Q is MgBr, MgCl, MgI or Li. 12. The method of claim 11, wherein the reduction step (B1) comprises contacting the compound of formula IIIc with a ketone reductase. 13. The method of claim 10, wherein the compound of formula IIId is prepared by the steps of: A2)Converting compound IIIf to form compound IIIg: (Claims 3 / 11, page 4, CN 122270438 A); and (B2) reacting said compound IIIg (e.g., esterification) to form said compound IIId: . 14. The method of claim 13, wherein said conversion step (A2) comprises forming a diazonium salt of said compound IIIf. 15. A method for preparing a compound of formula IV or a salt thereof: The method comprises the steps of: a) coupling a compound of formula III with a barium salt of compound A to form a compound of formula IVa; and b) hydrolyzing the compound of formula IVa to form the compound of formula IV: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 16. The method of claim 15, wherein the coupling step (a) comprises contacting the compound of formula III and the barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH). 17. A method for preparing a compound of formula VIII or a salt thereof: the method comprises the steps of: a) esterifying and N-alkylating a compound of formula VIIIa to form a compound of formula VIIIb; b) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers; and c) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 18. A method for preparing a compound of formula VIII or a salt thereof: The method comprises the following steps: Claims 5 / 11, page 6, CN 122270438 A a) esterifying a compound of formula VIIIa to form a compound of formula VIIId; b) N-alkylating said compound VIIId to form said compound VIIIb; c) reducing said compound VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers; and d) separating the mixture of transisomers of said compound VIIIc to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 19. A method for preparing a compound of formula IX or a salt thereof:The method comprises the following steps: a) borate the compound of formula VIII to form the compound of formula X: claim 6 / 11 page 7 CN 122270438 A; and b) couple the compound of formula X to the compound of formula XI to form the compound of formula XII: ; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; and R14 is an optionally substituted 3- to 6-membered cycloalkyl. 20. A method for preparing a compound of formula XIV or a salt thereof: The method comprises the following steps: a) reacting a compound of formula XV with a compound of formula XII to form a compound of formula XVI; b) removing PGa from the compound of formula XVI to form a compound of formula XVII; c) cyclizing the compound of formula XVII with a transition metal catalyst to form a compound of formula XVIII; d) removing PGa from the compound of formula XVIII to form a compound of formula XIX; and e) coupling the compound of formula XIX with the compound of formula IV to form a compound of formula XIV; wherein n and m are each independently 0, 1, 2, 3, 4 or 5; X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-; R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; PGa is an acid-instability protecting group; and PGb is an acid-stability protecting group. 21. The method of claim 20, wherein step (a) comprises reacting the compound of formula XII with the compound of formula XV in the presence of a carbodiimide coupling agent, an anti-racemic agent, and a base. 22. The method of claim 20 or 21, wherein step (c) further comprises contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII. 23. The method of any one of claims 20 to 22, wherein step (e) comprises coupling the compound of formula XIX with the compound of formula IV in the presence of a coupling agent, an anti-racemic agent, and a base. 24. A compound of formula IIf-1: or a salt thereof, wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl; R20 is hydrogen or a C1-6 alkyl group optionally substituted with one or two substituents independently selected from CN, OH, C1-6 alkoxy, and C1-6 haloalkyl; R21 is OH, halogen, CN, C1-6 alkyl, C1-6 alkoxy, or C1-6 haloalkyl; and q is 0, 1, or 2. 25. A method for preparing a compound of formula IIf:The method comprises alkylating a compound of formula IIe with a methylating agent: wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl. 26. The method of claim 25, wherein the methylating agent comprises trimethylcycloboroxane. 27. A method for preparing a compound of formula IVa: Claims 9 / 11, page 10, CN 122270438 A. The method comprises coupling a compound of formula III with a compound of formula IIf, wherein n and m are each independently 0, 1, 2, 3, 4, or 5; each R4 is independently an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; each R5 is independently an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl, or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-. 28. The method of claim 27, wherein the coupling comprises contacting the compound of formula III and the compound of formula IIf with a base. 29. A method for preparing a compound of formula VIIIc: The method comprises N-alkylating a compound of formula VIIIe with a compound having a structure having R13-LG1: wherein each R12 is a halogen; each R13 is an optionally substituted C1-C6 alkyl group; and LG1 is a detachable group. 30. A method for preparing a compound of formula VIIIe: Claims 10 / 11 pages 11 CN 122270438 A The method comprises reducing a compound of formula VIIIf with a reducing agent: wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 31. The method of claim 30, wherein the reducing agent is NaBH4. Claims 11 / 11 pages 12 CN 122270438 A Background Art for the Synthesis of Ras Inhibitors

[0001] Most small molecule drugs work by binding to functionally important pockets on target proteins, thereby modulating the activity of said proteins. For example, cholesterol-lowering drugs called statins bind to the active site of HMG-CoA reductase, thereby preventing the enzyme from binding to its substrate. In fact, many such drug / target interactions are known, which may mislead one into believing that small molecule regulators targeting most (if not all) proteins could be discovered given a reasonable amount of time, effort, and resources. This is far from the case. Currently, it is estimated that only about 10% of all human proteins are targetable by small molecules. (Bojadzic and Buchwald, Curr Top Med Chem 18: 674–699 (2019)). It is currently believed that the other 90% are targeted by small molecules.Small molecule drug discovery is often difficult or challenging. Such targets are often referred to as “undruggable.” These undruggable targets comprise a large portion of medically important human proteins and are often untapped reservoirs. Therefore, there is great interest in discovering novel molecular modalities that can modulate the function of such undruggable targets.

[0002] It is well established in the literature that Ras proteins (K-Ras, H-Ras, and N-Ras) play a crucial role in a variety of human cancers and are therefore suitable targets for anticancer therapies. In fact, in the United States, Ras protein mutations cause approximately 30% of all human cancers, many of which are fatal. Dysregulation of Ras proteins through activating mutations, overexpression, or upstream activation is common in human tumors, and activating mutations of Ras are frequently found in human cancers. For example, an activating mutation at codon 12 in the Ras protein significantly tilts the Ras mutant protein population to the "on" (GTP-binding) state (Ras(ON)) by inhibiting GTPase activator protein (GAP) dependence and intrinsic GTP hydrolysis rate, leading to oncogenic MAPK signaling. Notably, Ras exhibits a picomolar affinity for GTP, allowing it to be activated even in low concentrations of this nucleotide. Mutations at codons 13 (e.g., G13C) and 61 (e.g., Q61K) in Ras also induce oncogenic activity in some cancers.

[0003] Despite extensive drug discovery efforts against Ras over the past decades, only two agents targeting the K-Ras G12C mutant have been approved in the United States (sotorasib and adagrasib). More effort is needed to discover other medicines for cancers driven by various Ras mutations, and convenient, scalable synthetic methods remain required.

[0004] The present invention provides a method for preparing Ras inhibitors, an intermediate that can be used to synthesize Ras inhibitors, and a method for preparing said intermediates.

[0005] In one aspect, the present disclosure provides a method for separating enantiomers of a compound of formula I. The method includes the steps of: a) contacting a compound of formula I with a chiral acid to form a diastereomeric salt of the compound of formula I; and b) separating each diastereomeric salt of each diastereomeric isomer, thereby separating the enantiomers of the compound of formula I, wherein n and m are each independently 0, 1, 2, 3, 4, or 5; PG is a nitrogen protecting group; and X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

[0006] In some embodiments, the chiral acid is a carboxylic acid. In some embodiments, the chiral carboxylic acid is chiral.Dicarboxylic acid. In some embodiments, the chiral dicarboxylic acid is L(+)-tartaric acid. In some embodiments, the separation step (b) includes recrystallization. In some embodiments, the nitrogen protecting group is Boc (tert-butyloxycarbonyl).

[0007] In some embodiments, the compound of formula I is further described by the compound of formula Ia: .

[0008] In some embodiments, the compound of formula I or formula Ia is compound B: .

[0009] On the other hand, this disclosure provides a barium salt of compound A: .

[0010] In some embodiments, the barium salt is a 2:1 (compound A: barium) salt.

[0011] On the other hand, this disclosure provides a method for preparing a barium salt of compound A. The method comprises the following steps: a) condensing compound IIa with compound C to form compound IIb; b) cyclizing compound IIb with compound IIc to form compound IId; c) hydrolyzing compound IId to form compound IIe; d) methylating compound IIe to form compound IIf; and e) hydrolyzing compound IIf to form a barium salt of compound A: wherein R2 and R3 are each independently an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-C10 aryl, and LG is a detached group selected from the group consisting of halogen, trifluoromethanesulfonate, methanesulfonate, and toluenesulfonate.

[0012] In some embodiments of the method for preparing a barium salt of compound A, the condensation step (a) comprises contacting compound IIa and compound C with a base. In some embodiments, the cyclization step (b) includes contacting compounds of formula IIb and IIc with a base. In some embodiments, the hydrolysis step (c) includes contacting compound IId with an acid. In some embodiments, the methylation step (d) includes contacting compound IIe with MeB(OH)₂. In some embodiments, the hydrolysis step (e) includes contacting compound IIf with barium hydroxide. In some embodiments, R3 is an optionally substituted C6-C10 aryl group. In some embodiments, R3 is... In some embodiments, R3 is an optionally substituted C1-C6 alkyl group. In some embodiments, R3 is tert-butyl. In some embodiments, R2 is an optionally substituted C1-C6 alkyl group, an optionally substituted C3-6 cycloalkyl group, or an optionally substituted C6-10 aryl group. In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, LG is a halogen. In some embodiments, LG is -Br.

[0013] On the other hand, this disclosure provides a method for preparing a compound of formula III or a salt thereof: The method includes the following steps: Specification 3 / 123 pages 15 CN 122270438A) Couple the compound of formula IIId with the compound of formula I to form the compound of formula IIIe: ; and b) Deprotect the compound of formula IIIe to form the compound of formula III: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; each R4 is independently an optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl; and R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0014] In some embodiments, the nitrogen protecting group is Boc (tert-butoxycarbonyl). In some embodiments, the IIId compound is prepared by a method comprising the following steps: A1) combining the IIIa compound with the IIIb compound to form the IIIc compound; B1) reducing the IIIc compound to form the IIId compound.

[0015] Wherein Q is MgBr, MgCl, MgI or Li.

[0016] In some embodiments, the reduction step (B1) comprises contacting the IIIc compound with a ketone reductase.

[0017] In some embodiments, the IIId compound is prepared by a method comprising the following steps: A2) converting the IIIf compound to form the IIIg compound; and B2) reacting the IIIg compound (e.g., esterification) to form the IIId compound. In some embodiments, the conversion step (A2) comprises forming a diazonium salt of the IIIf compound.

[0018] In some embodiments, step (b) comprises contacting the IIIe compound with an acid.

[0019] In some embodiments, the compound of formula III is compound D: The compound is prepared by the following steps: a) coupling the compound of formula IIId-1 with compound B to form the compound of formula IIIe-1 having the following structure: ; and b) deprotecting the compound of formula IIIe-1 to form compound D having the following structure: .

[0020] On the other hand, this disclosure provides a method for preparing a compound of formula IV or a salt thereof: Specification 5 / 123 pages 17 CN 122270438 A.

[0021] The method includes the following steps: a) coupling the compound of formula III with a barium salt of compound A to form a compound of formula IVa: ; and b) hydrolyzing the compound of formula IVa to form a compound of formula IV: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl;R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

[0022] In some embodiments, the coupling step (a) includes contacting the compound of formula III and the barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH). In some embodiments, after step (a) is completed, the compound of formula IVa in the form of maleate is isolated. In some embodiments, after step (a) is completed, the compound of formula IVa in the form of benzenesulfonate is isolated. In some embodiments, the hydrolysis step (b) includes contacting the compound of formula IVa with an acid or base. In some embodiments, the hydrolysis step (b) includes hydrogenating the compound of formula IVa. In some embodiments, hydrogenation further includes contacting the compound of formula IVa with a palladium catalyst.

[0023] In some embodiments, the method includes preparing compound E: Specification 6 / 123 page 18 CN 122270438 A and the method includes the steps of: a) coupling compound D with a barium salt of compound A to form a compound of formula IVa having the following structure: ; and b) deprotecting the compound of formula IVa to form compound E having the following structure: .

[0024] In some embodiments, after step (a), the compound of formula IVa in the form of maleate is isolated. In some embodiments, after step (a), the compound of formula IVa in the form of benzenesulfonate is isolated.

[0025] In some embodiments of any methods and compounds described herein, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0026] In some embodiments of any methods and compounds described herein, each of X1, X2 and X3 is CH2. In some embodiments, PG is . In some embodiments, LG is a halogen. In some embodiments, LG is Br.

[0027] In some embodiments, R3 is an optionally substituted C6-C10 aryl. In some embodiments, R3 is... In some embodiments, R3 is an optionally substituted C1-C6 alkyl. In some embodiments, R3 is... In some embodiments, R2 is an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl. In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, R4 is optionally...The alkyl group is a C1-C6 alkyl group or an optionally substituted C6-C10 aryl group. In some embodiments, R4 is benzyl, methyl, or tert-butyl. In some embodiments, R5 is an optionally substituted C1-C6 alkyl group or an optionally substituted 3- to 6-membered cycloalkyl group. In some embodiments, R5 is an optionally substituted C1-C6 alkyl group. In some embodiments, R5 is isopropyl. In some embodiments, R5 is an optionally substituted 3- to 6-membered cycloalkyl group. In some embodiments, R5 is cyclopentyl.

[0028] On the other hand, this disclosure provides a method for preparing a compound of formula VIII or a salt thereof: (Page 7 / 123, CN 122270438 A) The method comprises the following steps: a) esterifying and N-alkylating a compound of formula VIIIa to form a compound of formula VIIIb; b) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers; and c) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0029] On the other hand, this disclosure provides a method for preparing a compound of formula VIII-1 or a salt thereof: the method includes the following steps: a) esterifying and N-alkylating a compound of formula VIIIa-1 to form a compound of formula VIIIb-1: Specification 8 / 123 pages 20 CN 122270438 A; b) reducing the compound of formula VIIIb-1 to form a compound of formula VIIIc-1 in the form of a mixture of transisomers: ; and c) separating the mixture of transisomers of formula VIIIc-1 to obtain a stereochemically pure compound of formula VIII-1; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl; and p is 0, 1, 2 or 3.

[0030] In some embodiments, the compound of formula VIII is compound F: and the method includes the following steps: a) esterifying and N-alkylating the compound of formula VIIIa to form a compound of formula VIIIb having the following structure: b) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers having the following structure: and c) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII.

[0031] On the other hand, this disclosure provides a method for preparing a compound of formula VIII or a salt thereof: the method includes the following steps: a)a) Reacting compound VIIIa (e.g., esterification) to form compound VIIId; b) N-alkylating compound VIIId to form compound VIIIb; c) Reducing compound VIIIb to form compound VIIIc in the form of a mixture of transisomers: (See specification 10 / 123, page 22, CN 122270438 A); and d) Separating the mixture of transisomers of VIIIc to obtain a stereochemically pure compound VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0032] On the other hand, this disclosure provides a method for preparing a compound of formula VIII-1 or a salt thereof: the method includes the following steps: a) reacting a compound of formula VIIIa-1 (e.g., esterification) to form a compound of formula VIIId-1; b) N-alkylating a compound of formula VIIId-1 to form a compound of formula VIIIb-1; c) reducing a compound of formula VIIIb-1 to form a compound of formula VIIIc-1 in the form of a mixture of transisomers: (See specification 11 / 123 pages 23 CN 122270438 A); and d) separating the mixture of transisomers of formula VIIIc-1 to obtain a stereochemically pure compound of formula VIII-1; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; Each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl; and p is 0, 1, 2, or 3.

[0033] In some embodiments, the compound of formula VIII is compound G: and the method includes the following steps: a) reacting the compound of formula VIIIa (e.g., esterification) to form a compound of formula VIIId having the following structure: b) N-alkylating the compound of formula VIIId to form a compound of formula VIIIb having the following structure: ; c) reducing the compound of formula VIIIb to form compound G in the form of a mixture of transisomers having the following structure: Specification 12 / 123 pages 24 CN 122270438 A ; and d) separating the mixture of transisomers of compound G to obtain a stereochemically pure compound G.

[0034] In some embodiments, the mixture of transisomers is separated by column chromatography or chemical resolution. In some embodiments, chemical resolution involves contacting the compound of formula VIIIc with an acid. In some embodiments, the acid is methanesulfonic acid. In some embodiments, the acid is camphorsulfonic acid (e.g., D-camphor-10-sulfonic acid). In some embodiments,Step (b) includes contacting compound VIIIa or compound VIIId with 2,2,2-trifluoroethyl trifluoromethanesulfonate: .

[0035] In some embodiments, step (c) includes contacting compound VIIIb with lithium borohydride (LiBH4).

[0036] On the other hand, this disclosure provides a method for preparing compound IX or a salt thereof: the method includes the steps of: a) borate compound VIII to form compound X: ; and b) couple compound X with compound XI to form compound XII: Specification 13 / 123 pages 25 CN 122270438 A; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; and R14 is an optionally substituted 3- to 6-membered cycloalkyl.

[0037] On the other hand, this disclosure provides a method for preparing a compound of formula IX-1 or a salt thereof: the method includes the following steps: a) borate a compound of formula VIII-1 to form a compound of formula X-1: ; and b) couple a compound of formula X-1 with a compound of formula XI to form a compound of formula XII-1: ; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; R14 is an optionally substituted 3- to 6-membered cycloalkyl; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocyclic alkyl; and p is 0, 1, 2 or 3.

[0038] In some embodiments, the compound of formula IX is compound H: The method includes the steps of: a) borate the compound of formula VIII to form a compound of formula X having the following structure: ; and b) couple the compound of formula X with the compound of formula XI to form a compound H having the following structure: .

[0039] In some embodiments, the borate step (a) includes contacting the compound of formula VIII with an iridium catalyst. In some embodiments, the coupling step (b) includes contacting the compound of formula X and the compound of formula XI with a copper source and a base. In some embodiments, the copper source is Cu(OAc)2. In some embodiments, the base is triethylamine or tetramethylpiperidine.

[0040] In some embodiments, the compound of formula X or a salt thereof: Specification 15 / 123, page 27, CN 122270438 A is further prepared by the following steps: a) boronizing the compound of formula VIII to form the compound of formula XIII: ; and b) hydrolyzing and boronizing the compound of formula XIII in one step to form formula X: ; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0041] In some embodiments, the compound of formula X-1 or a salt thereof is further prepared by the following steps: a) boronizing the compound of formula VIII-1 to form the compound of formula XIII-1: (See specification 16 / 123, page 28, CN 122270438 A); and b) hydrolyzing and boronizing the compound of formula XIII-1 in one step to form formula X-1: ; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl; and p is 0, 1, 2, or 3.

[0042] On the other hand, this disclosure provides a method for preparing a compound of formula XIV or a salt thereof: the method includes the following steps: a) reacting a compound of formula XV with a compound of formula XII (e.g., esterification) to form a compound of formula XVI: (Specification 17 / 123, page 29, CN 122270438 A); b) removing (e.g., deprotecting) PGa of the compound of formula XVI to form a compound of formula XVII; c) cyclizing the compound of formula XVII with a transition metal catalyst to form a compound of formula XVIII; d) removing (e.g., deprotecting) PGa of the compound of formula XVIII to form a compound of formula XIX; and e) coupling the compound of formula XIX with a compound of formula IV to form a compound of formula XIV: (Specification 18 / 123, page 30, CN 122270438 A); wherein n and m are each independently 0, 1, 2, 3, 4 or 5; X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-; R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; PGa is an acid-instability protecting group; and PGb is an acid-stability protecting group.

[0043] In some embodiments of step (c), the transition metal catalyst includes palladium, platinum, nickel, copper, and iron catalysts. In some embodiments, the transition metal catalyst is a palladium catalyst.

[0044] In some embodiments, this disclosure provides a method for preparing a compound of formula XVI. The method includes reacting a compound of formula XV with a compound of formula XII under conditions sufficient to form a compound of formula XVI. The compound of formula XV can be prepared according to the process described herein.

[0045] In some embodiments, this disclosure provides a method for preparing a compound of formula XVIII. The method includes cyclizing a compound of formula XVII in the presence of a transition metal catalyst (such as a palladium catalyst) to form a compound of formula XVIII. Compounds of formula XVII can be prepared according to the process described herein.

[0046] In some embodiments, this disclosure provides a method for preparing a compound of formula XIV. The method includes reacting a compound of formula XIX with a compound of formula IV under conditions sufficient to form a compound of formula XIV. The compounds of formula XIX and IV can be prepared according to the process described herein.

[0047] On the other hand, this disclosure provides a method for preparing a compound of formula XIV-1 or a salt thereof: the method includes the following steps: a) reacting a compound of formula XV with a compound of formula XII-1 (e.g., esterification) to form a compound of formula XVI-1: Specification 19 / 123 pages 31 CN 122270438 A; b) removing (e.g., deprotecting) PGa of the compound of formula XVI-1 to form a compound of formula XVII-1; c) cyclizing the compound of formula XVII-1 with a transition metal catalyst to form a compound of formula XVII-1; d) removing (e.g., deprotecting) PGa of the compound of formula XVIII-1 to form a compound of formula XIX-1; and e) coupling the compound of formula XIX-1 with a compound of formula IV to form a compound of formula XIV-1: Specification 20 / 123 pages 32 CN 122270438 A; wherein n and m are each independently 0, 1, 2, 3, 4 or 5; X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-; R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl; R14 is an optionally substituted 3- to 6-membered cycloalkyl; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocyclic alkyl; p is 0, 1, 2, or 3; PGa is an acid-instability protecting group; and PGb is an acid-stability protecting group.

[0048] In some embodiments of step (c), the transition metal catalyst includes palladium, platinum, nickel, copper, and iron catalysts. In some embodiments, the transition metal catalyst is a palladium catalyst.

[0049] In some embodiments, the compound of formula XIV is compound 1: Compound 1, and the method includes the following steps: a) reacting the compound of formula XV with the compound of formula XII (e.g., esterification) to form the compound of formula XVI having the following structure: ; b) deprotecting the Boc of the compound of formula XVI to form the compound of formula XVII having the following structure: ; c) cyclizing the compound of formula XVII with a transition metal catalyst to form the compound of formula XVII having the following structure: ; d) removing (e.g., deprotecting) the PGb of the compound of formula XVIII to form the compound of formula XIX having the following structure: ; and e)The compound of formula XIX is coupled with the compound of formula IV to form compound 1 having the following structure: .

[0050] In some embodiments, step (a) includes contacting the compound of formula XII with the compound of formula XV in the presence of a carbodiimide coupling agent, an anti-racemic agent, and a base. In some embodiments, the carbodiimide coupling agent is EDCl, the anti-racemic agent is HOBt, and the base is N,N-diisopropylethylamine. In some embodiments, the deprotection step (b) further includes contacting the compound of formula XVI with an acid. In some embodiments, the acid is trifluoroacetic acid. In some embodiments, the transition metal catalyst in step (c) includes palladium, platinum, nickel, copper, and iron catalysts. In some embodiments, the transition metal catalyst is a palladium catalyst. In some embodiments, the cyclization step (c) further includes contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII. In some embodiments, the deprotection step (d) further includes washing the compound of formula XIX with N-acetylcysteine. In some embodiments, step (e) includes coupling the compound of formula XIX to the compound of formula IV in the presence of a coupling agent, an anti-racemic agent, and a base. In some embodiments, the coupling agent is PyBOP or EDCl. In some embodiments, the anti-racemic agent is selected from the group consisting of Oxyma, HOBt, or HOPO. In some embodiments, the base is N,N-diisopropylethylamine. In some embodiments, the method further includes a step of purifying the compound of formula XIV. In some embodiments, purification includes recrystallizing the compound of formula XIV. In some embodiments, recrystallization includes adding a first solvent, followed by adding a second solvent. In some embodiments, the first solvent is dioxane or 2-methyltetrahydrofuran. In some embodiments, the second solvent is diisopropyl ether or isopropanol. In some embodiments, recrystallization further includes adding a third solvent. In some embodiments, the third solvent is heptane. In some embodiments, R12 is -Br. In some embodiments, R13 is -CH2CF3. In some embodiments, PGa is Boc. In some embodiments, PGb is CBz.

[0051] On the other hand, this disclosure provides a compound of formula IIf-1: or a salt thereof, wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl; R20 is hydrogen or a C1-6 alkyl optionally substituted with one or two substituents independently selected from CN, OH, C1-6 alkoxy, and C1-6 haloalkyl; R21 is OH, halogen, CN, C1-6 alkyl, C1-6 alkoxy, or C1-6 haloalkyl; and q is 0, 1, or 2.

[0052] In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, R2 is benzyl.

[0053] In some embodiments, the compound of formula IIf-1 has the structure of formula IIe or a salt thereof: .

[0054] In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, R2 is benzyl.

[0055] In some embodiments, the compound of formula IIf-1 has the structure of formula IIf or a salt thereof: Specification 23 / 123 pages 35 CN 122270438 A .

[0056] In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, R2 is benzyl.

[0057] On the other hand, this disclosure provides a method for preparing a compound of formula IIf: The method includes alkylating a compound of formula IIe with a methylating agent: wherein R2 is an optionally substituted C1-6 alkyl, an optionally substituted C3-6 cycloalkyl, or an optionally substituted C6-10 aryl.

[0058] In some embodiments, the methylating agent comprises trimethylcycloboroxane. In some embodiments, the methylating agent comprises trimethylcycloboroxane and Cu(OAc)2. In some embodiments, the methylating agent comprises trimethylcycloboroxane and CuOAc.

[0059] On the other hand, this disclosure provides a method for preparing a compound of formula IVa: the method comprises coupling a compound of formula III with a compound of formula IIf, wherein n and m are each independently 0, 1, 2, 3, 4 or 5; each R4 is independently an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; each R5 is independently an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0060] In some embodiments, R4 is benzyl, methyl, or tert-butyl.

[0061] In some embodiments, R5 is isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, R5 is cyclopentyl.

[0062] In some embodiments, coupling includes contacting the compound of formula III and the compound of formula IIf with a base. In some embodiments, the base is 1,5,7-triazabicyclo[4,4,0]dec-5-ene (TBD). Specification 24 / 123 pages 36 CN 122270438 A

[0063] On the other hand, this disclosure provides a method for preparing a compound of formula VIIIc: the method includes N-alkylating a compound of formula VIIIe with a compound having a structure having R13-LG1: wherein each R12 is independently a halogen; each R13 is independently an optionally substituted C1-C6 alkyl group; and LG1 is a detachable group.

[0064] In some embodiments, each R12 is Br.

[0065] In some embodiments, each R13 is -CH2CF3.

[0066] In some embodiments, LG1 is a halogen, trifluoromethanesulfonate group, methanesulfonate group, or toluenesulfonate group.

[0067] In some embodiments, R13-LG is 2,2,2-trifluoroethyl trifluoromethanesulfonate.

[0068] On the other hand, this disclosure provides a method for preparing a compound of formula VIIIe: the method includes reducing the compound of formula VIIIf with a reducing agent: wherein R12 is a halogen; and R13 is optionally substituted C1-C6 alkyl.

[0069] In some embodiments, each R12 is Br.

[0070] In some embodiments, each R13 is -CH2CF3.

[0071] In some embodiments, the reducing agent is NaBH4.

[0072] Definitions and Chemical Terms In this application, unless clearly indicated from the context, (i) the term “a” means “one or more”; (ii) the term “or” is used to mean “and / or” unless explicitly indicated that the term indicates that the alternative is unique or that the alternatives are mutually exclusive, however, this disclosure supports the definition of indicating a unique alternative and “and / or”; (iii) the terms “comprising” and “including” should be understood to encompass the listed components or steps, whether presented alone or together with one or more additional components or steps; and (iv) when providing a range, endpoints are included.

[0073] As used herein, the term “about” is used to indicate that a value includes the standard deviation of the error of the means or method used to determine the value. In some embodiments, unless otherwise specified or otherwise obvious from the context (e.g., such figures would exceed 100% of possible values), the term “about” refers to a range of values ​​in any direction (greater than or less than) within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the stated value.

[0074] As used herein, in the context of describing adjacent atoms, the term “adjacent” refers to a divalent atom directly connected by a covalent bond.

[0075] As used herein, whether explicitly indicated or not, “compound of the invention” and similar terms refer to the Ras inhibitor described herein (e.g., compound A) and intermediates in its synthesis, as well as its salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including transisomers), and tautomers.

[0076] As used herein, the term "nitrogen protecting group" refers to a removable chemical group on the nitrogen atom of the compounds described herein.Part of it protects nitrogen from unwanted side reactions. Non-limiting examples of nitrogen protecting groups include Boc (tert-butyloxycarbonyl), Fmoc (fluorenylmethoxycarbonyl), and Cbz (benzyl carbamate).

[0077] Those skilled in the art will understand that some of the compounds described herein may exist in one or more different isomers (e.g., stereoisomers, geometric isomers, transisomers, tautomers) or isotopic forms (e.g., where one or more atoms have been substituted with different isotopes of said atoms, such as hydrogen-substituted deuterium). Unless otherwise indicated or clearly apparent from the context, the illustrated structures are to be understood as individually or in combination representing any such isomer or isotopic form.

[0078] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are contemplated. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic form. Methods for preparing optically active forms from optically inactive starting materials are known in the art, such as by the resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of alkenes, C=N double bonds, etc., may also be present in the compounds described herein, and all such stable isomers are covered in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described and can be isolated as mixtures of isomers or separate isomeric forms.

[0079] In some embodiments, one or more compounds illustrated herein may exist in different tautomeric forms. It will be clear from the context that, unless explicitly excluded, references to such compounds cover all such tautomeric forms. In some embodiments, the tautomeric form is obtained by exchanging a single bond with an adjacent double bond and accompanied by proton migration. In some embodiments, the tautomeric form may be a proton-transfer tautomeric form, which is an isomeric protonated state having the same empirical formula and total charge as the reference form. Examples of portions having proton transfer tautomer forms (pages 26 / 123, 38, CN 122270438 A) include keto-enol pairs, amide-imine pairs, lactam-lactamimide pairs, amide-imine pairs, enamine-imine pairs, and cyclic forms, in which the proton may occupy two or more positions in the heterocyclic system, such as 1H-imidazolium and 3H-imidazolium, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. In some embodiments, the tautomer forms may be in equilibrium or spatially locked into one form by appropriate substitution. In some embodiments, the tautomer forms are obtained by interconversion of acetals.

[0080] Unless otherwise stated, the structures illustrated herein are also intended to include compounds distinguished only by the presence of one or more isotopically enriched atoms. Exemplary isotopes that may be incorporated into the compounds of this invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 32P, 33P, 35S, 18F, 36Cl, 123I, and 125I. Isotope-labeled compounds (e.g., those labeled with 3H and 14C) can be used for compound or substrate tissue distribution analysis. Tritium (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are available due to their ease of preparation and detectability. Furthermore, substitution with a heavier isotope such as deuterium (i.e., 2H) can provide certain therapeutic advantages due to higher metabolic stability (e.g., increased in vivo half-life or reduced dose requirement). In some embodiments, one or more hydrogen atoms are substituted with 2H or 3H, or one or more carbon atoms are substituted with carbon enriched with 13C or 14C. Positron emission isotopes (such as 15O, 13N, 11C, and 18F) can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. The preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can generally be prepared by following procedures similar to those disclosed with respect to the compounds of the invention described herein, by substituting a non-isotopically labeled reagent with an isotopically labeled reagent.

[0081] As is known in the art, many chemical entities can be in a variety of different solid forms, such as amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the invention can be used in any of these forms, including any solid form. In some embodiments, the compounds described or illustrated herein may be provided or used in hydrate or solvate form.

[0082] At various locations throughout this specification, substituents of the compounds disclosed herein are disclosed in either group or range form. This disclosure is specifically intended to include each individual combination of members of such groups and ranges. For example, the term “C1-C6 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl. Furthermore, where a compound comprises multiple positions, and at said positions, substituents are disclosed in either group or range form, unless otherwise indicated, this disclosure is intended to cover individual compounds and groups (e.g., genera and subgenera) containing each individual combination of members at each position.

[0083] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein the alkyl is optionally substituted”). It is not intended that the feature “X” (e.g., alkyl) itself is anyOptional. As described herein, certain compounds of interest may contain one or more “optionally substituted” moieties. Generally, the term “substituted” (whether preceded by the term “optionally”) means that one or more hydrogens of the specified moieties are replaced by suitable substituents (e.g., any substituents or groups described herein). Unless otherwise indicated, “optionally substituted” groups may have suitable substituents at each substituted position of the group, and the substituents at each position may be the same or different when more than one position in any given structure may be substituted by more than one substituent selected from the prescribed group. For example, in the term “optionally substituted C1-C6 alkyl-C2-C9 heteroaryl,” the alkyl moieties, the heteroaryl moieties, or both may be optionally substituted. The combinations of substituents contemplated in this disclosure are preferably those that result in the formation of stable or chemically viable compounds. As used herein, the term “stable” means that a compound remains substantially unchanged when subjected to conditions that allow it to be generated, detected, and in some embodiments, recovered, purified, and used for one or more of the purposes disclosed herein. Specification 27 / 123 Page 39 CN 122270438 A

[0084] Suitable monovalent substituents on the substituted carbon atoms of the “optionally substituted” group can be independently deuterium; halogen; -(CH2)O-4R o; -(CH2)O-4OR o; -O(CH2)O-4R o; -O-(CH2)O-4C(O)OR o; -(CH2)O-4CH(OR o)2; -(CH2)O-4SR o; -(CH2)O-4Ph, which can be substituted by Ro; -(CH2)O-4O(CH2)O-1Ph, which can be substituted by Ro; -CH=CHPh, which can be substituted by Ro Substitution; -(CH2)0-4O(CH2)0-1-pyridyl, which may be substituted with Ro; 4-8 membered saturated or unsaturated heterocyclic alkyl (e.g., pyridyl); 3-8 membered saturated or unsaturated cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl); -NO2; -CN; -N3; ​​-(CH2)0-4N(Ro)2; -(CH2)0-4N(Ro)C(O)Ro; -N(Ro)C(S)Ro; -(CH2)0-4N(Ro)C(O)NRo2; -N(Ro)C(S)NRo2; -(CH2)0-4N(Ro)C(O)ORo; -N(Ro)N(Ro)C(O)Ro; -N(Ro)N(Ro)C(O)NRo2; -N(Ro)N(Ro)C(O)ORo; -(CH2)0-4C(O)R o ;‑C(S)Ro;‑(CH2)0‑4C(O)OR o ;‑(CH2)0‑4‑C(O)‑N(R o)2;‑(CH2)0‑4‑C(O)‑N(R o)‑ S(O)2‑R o;‑C(NCN)NRo2;‑(CH2)0‑4C(O)SR o ;‑(CH2)0‑4C(O)OsiR o 3;‑(CH2)0‑4OC(O)R o ;‑OC(O) (CH2)0‑4SR o ;‑SC(S)SRo ;‑(CH2)0‑4SC(O)R o ;‑(CH2)0‑4C(O)NR o 2;‑C(S)NR o 2;‑C(S)SR o ;‑(CH2)0‑4OC(O)NR o 2;‑C(O)N(OR o)Ro;‑C(O)C(O)Ro;‑C(O)CH2C(O)R o ;‑C(NORo)Ro;‑(CH2)0‑ 4SSR o ;‑(CH2)0‑4S(O)2R o ;‑(CH2)0‑4S(O)2OR o ;‑(CH2)0‑4OS(O)2R o ;‑S(O)2NR o 2;‑(CH2)0‑4S(O) Ro;‑N(Ro)S(O)2NR o 2;‑N(R o)S(O)2R o ;‑N(ORo)Ro;‑C(NORo)NRo2;‑C(NH)NR o 2;‑P(O)2R o;‑P(O) Ro2;‑P(O)(OR o)2;‑OP(O)R o 2;‑OP(O)(OR o)2;‑OP(O)(OR o)Ro, -SiRo3; -(C1-4 straight-chain or branched-chain alkylene)O-N(Ro)2; or -(C1-4 straight-chain or branched-chain alkylene)C(O)O-N(Ro)2, wherein each Ro may be substituted as defined below and is independently hydrogen, -C1-6 aliphatic group, -CH2Ph, -O(CH2)0-1Ph, -CH2-(5-6 membered heteroaryl ring) or a 3-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or notwithstanding the foregoing definition, two independently occurring Ros together with their intervening atoms form a 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0085] Suitable monovalent substituents on Ro (or a ring formed by two independently occurring Ro atoms and their intervening atoms) can be independently halogens, -(CH2)O-2R, -(halogenated R), -(CH2)O-2OH, -(CH2)O-2OR, -(CH2)O-2CH(OR)2; -O (halogenated R), -CN, -N3, -(CH2)O-2C(O)R, -(CH2)O-2C(O)OH, -(CH2)O-2C(O)OR, -(CH2)O-2SR, -(CH2)O-2SH, -(CH2)O-2NH2, -(CH2)O-2NHR● -(CH2)0-2NR ● 2 -NO2 -SiR ● 3 -OsiR ● 3 -C(O)SR ● - (C1-4 straight-chain or branched-chain alkylene)C(O)OR● or -SSR●, wherein each R● is unsubstituted or, if preceded by “halogenated”, substituted by only one or more halogens, and is independently selected from C1-4 aliphatic groups, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of Ro include =O and =S.

[0086] Suitable divalent substituents on the saturated carbon atom of the “optionally substituted” group include the following: =O, =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR*, =NOR*, -O(C(R*2))2-3O- or -S(C(R*2))2-3S-, wherein each independently occurring R* is selected from hydrogen, a C1-6 aliphatic group that may be substituted as defined below, or an unsubstituted 5-6 saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that bond to adjacent substituted carbons of the "optionally substituted" group include: -O(CR*2)2-3O-, wherein each independently occurring R* is selected from hydrogen, a C1-6 aliphatic group that may be substituted as defined below, or an unsubstituted 5-6 saturated, partially unsaturated, or aryl ring having 0-4 independently selected heteroatoms selected from nitrogen, oxygen, or sulfur.

[0087] Suitable substituents on the aliphatic group of R* include halogen, -R●, -(halogenated R●), -OH, -OR●, -O(halogenated R●), -CN, -C(O)OH, -C(O)OR●, -NH2, -NHR●, -NR●2 or -NO2, wherein each R● is unsubstituted or, when preceded by "halogenated", is substituted by only one or more halogens, and is independently a C1-4 aliphatic group, -CH2Ph, -O(CH2)0-1Ph or a 5-6 member saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0088] Suitable substituents on the substituted nitrogen of the “optionally substituted” group include -R†, -NR†2, -C(O)R†, -C(O)C(O)R†, -C(O)CH2C(O)R†, -S(O)2R†, -S(O)2NR†2, -C(S)NR†2, -C(NH)NR†2 or -N(R†)S(O)2R†; wherein each R† is independently hydrogen; a C1-6 aliphatic group, which may be substituted as defined below; an unsubstituted -OPh; or havingAn unsubstituted 3-6 saturated, partially unsaturated, or aryl ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen, or sulfur, or, notwithstanding the foregoing definition, two independently occurring R† together with their intermediary atoms to form an unsubstituted 3-12 saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen, or sulfur.

[0089] Suitable substituents on the aliphatic group of R† are independently halogens, -R●, -(halogenated R●), -OH, -OR●, -O(halogenated R●), -CN, -C(O)OH, -C(O)OR●, -NH2, -NHR●, -NR●2, or -NO2, wherein each R● is unsubstituted or, when preceded by "halogenated", is substituted by only one or more halogens, and is independently a C1-4 aliphatic group, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of R† include =O and =S.

[0090] As used herein, the term "acetyl" refers to the group -C(O)CH3.

[0091] As used herein, the term "alkoxy" refers to an -O-C1-C20 alkyl group, wherein the alkoxy group is attached to the remainder of the compound via an oxygen atom.

[0092] As used herein, the term "alkyl" refers to a saturated, straight-chain or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms (e.g., 1 to 10 or 1 to 6). In some embodiments, the alkyl group is unbranched (i.e., straight-chain); in some embodiments, the alkyl group is branched. Examples of alkyl groups are, but are not limited to, methyl, ethyl, n-propyl and isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl, and neopentyl.

[0093] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group derived from a straight-chain or branched saturated hydrocarbon by removing two hydrogen atoms, and examples are methylene, ethylene, isopropylene, etc. The term "Cx-Cy alkylene" refers to an alkylene group having carbon atoms between x and y. Exemplary values ​​for x are 1, 2, 3, 4, 5, and 6, and exemplary values ​​for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-C6, C1-C10, C2-C20, C2-C6, C2-C10, or C2-C20 alkylene groups). In some embodiments, the alkylene group may be further substituted with 1, 2, 3, or 4 substituents as defined herein.

[0094] As used herein, the term "alkenyl" refers to a monovalent straight-chain or branched chain group containing one or more carbon-carbon double bonds (unless otherwise specified) of 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons), and examples of such groups are vinyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyl groups include both cis and trans isomers.As used herein, the term "alkenyl" refers to a divalent straight-chain or branched-chain group containing 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) with one or more carbon-carbon double bonds (unless otherwise specified).

[0095] As used herein, the term "alkynyl" refers to a monovalent straight-chain or branched-chain group containing 2 to 20 carbons (e.g., 2 to 4, 2 to 6 or 2 to 10 carbons) with carbon-carbon triple bonds, and examples of such groups are ethynyl and 1-propynyl.

[0096] As used herein, the term "amino" refers to -N(R†)2, such as -NH2 and -N(CH3)2.

[0097] As used herein, the term "aminoalkyl" refers to an alkyl moiety substituted with one or more amino moieties on one or more carbon atoms.

[0098] As used herein, the term "aryl" refers to a monovalent monocyclic, bicyclic, or polycyclic system formed of carbon atoms, wherein the rings attached to the side groups are aromatic. Examples of aryl groups are phenyl, naphthyl, phenanthryl, and anthracene. The aryl ring may be attached to its side group at any heteroatom or carbocyclic atom that produces a stable structure, and any ring atom may optionally be substituted unless otherwise specified.

[0099] As used herein, the term “CO” indicates a bond. For example, a portion of the term -N(C(O)-(CO-C5 alkylene-H)- includes -N(C(O)-(CO alkylene-H)-, which is also represented as -N(C(O)-H)-.

[0100] As used herein, the terms “carbocyclic” and “carbocyclic group” refer to a monovalent, optionally substituted C3-C12 monocyclic, bicyclic, or tricyclic ring. (Specification 29 / 123 pages 41 CN 122270438 A) A ring structure, which may be a bridging ring, a fused ring, or a spiro ring, wherein all rings are formed of carbon atoms and at least one ring is non-aromatic. Carbocyclic structures include cycloalkyl, cycloalkenyl, and cycloynyl groups. Examples of carbocyclic groups are cyclohexyl, cyclohexenyl, cyclooctynyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indenyl, decahydronaphthyl, etc. The carbocyclic ring may be attached to its side group at any ring atom that produces a stable structure, and unless otherwise specified, any ring atom may optionally be substituted.

[0101] As used herein, the term “carbonyl” means a C(O) group, which may also be represented as C=O.

[0102] As used herein, the term “carboxyl” means -CO2H, (C=O)(OH), COOH, or C(O)OH, or its unprotonated counterpart.

[0103] As used herein, the term “cyano” means a -CN group.

[0104] As used herein, the term "cycloalkyl" refers to a monovalent saturated cyclic hydrocarbon group, which, unless otherwise specified, can be a bridging ring, fused ring, or spiro ring having three to eight ring carbons, and examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cycloheptyl.

[0105] As used herein, the term "cycloalkenyl" means a monovalent, non-aromatic saturated cyclic hydrocarbon group, which, unless otherwise specified, can be a bridging ring, fused ring, or spiro ring having three to eight cyclic carbons and containing one or more carbon-carbon double bonds.

[0106] As used herein, the term "diastereomer" means stereoisomers that are not mirror images of each other and are not superimposed on each other.

[0107] As used herein, the term "enantiomer" means each individual optically active form of the compound of the present invention having at least 80% (i.e., at least 90% of one enantiomer and at most 10% of the other enantiomer), preferably at least 90%, and more preferably at least 98% optical purity or enantiomer excess (as determined by standard methods in the art).

[0108] As used herein, the term "haloacetyl" means an acetyl group in which at least one hydrogen atom has been replaced by a halogen.

[0109] As used herein, the term “halogenated alkyl” means an alkyl moiety on one or more carbon atoms substituted with one or more identical or different halogen moieties.

[0110] As used herein, the term “halogen” means a halogen selected from bromine, chlorine, iodine, or fluorine.

[0111] As used herein, the term “heteroalkyl” means “alkyl” as defined herein, wherein at least one carbon atom has been replaced by a heteroatom (e.g., O, N, or S atom). The heteroatom may appear in the middle or at the end of the group.

[0112] As used herein, the term “heteroaryl” means a monovalent, monocyclic, or polycyclic structure containing at least one fully aromatic ring: that is, it contains 4n+2 π electrons in the monocyclic or polycyclic system and contains at least one heteroatom selected from N, O, or S in the aromatic ring. Exemplary unsubstituted heteroaryl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons. The term "heteroaryl" includes bicyclic, tricyclic, and tetracyclic groups, wherein any of the aforementioned heteroaryl rings is fused to one or more aryl rings or carbocyclic rings (e.g., a benzene ring or a cyclohexane ring). Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindolyl. The heteroaryl ring may be attached to its side group at any ring atom that produces a stable structure, and any ring atom may optionally be substituted unless otherwise specified. In some embodiments, the heteroaryl group is substituted with 1, 2, 3, or 4 substituents.

[0113] As used herein, the term "heterocyclic alkyl" refers to a monovalent monocyclic, bicyclic, or polycyclic system, which may be bridged, fused, or spirocyclic, wherein at least one ring is non-aromatic and said non-aromatic ring contains one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Five-membered rings have zero to two double bonds, and six-membered and seven-membered rings...The member ring has zero to three double bonds. Exemplary unsubstituted heterocyclic alkyl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons. The term "heterocyclic alkyl" also refers to a heterocyclic compound having a bridging polycyclic structure, wherein one or more carbons or heteroatoms bridge two non-adjacent members in a monocyclic ring, such as a quinine cycloyl group. The term "heterocyclic alkyl" includes bicyclic, tricyclic, and tetracyclic groups, wherein any of the aforementioned heterocycles is fused with one or more aromatic rings, carbocyclic rings, heteroaromatic rings, or heterocycles (e.g., aryl rings, cyclohexane rings, cyclohexene rings, cyclopentane rings, cyclopentene rings, pyridine rings, or pyrrolidine rings). Examples of heterocyclic alkyl groups are pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridine, and decahydronaphthidyl. The heterocyclic alkyl ring may be attached to its side group at any ring atom that produces a stable structure, and any ring atom may optionally be substituted unless otherwise specified.

[0114] As used herein, the term "hydroxyl" means a -OH group.

[0115] As used herein, the term "hydroxyalkyl" means an alkyl moiety substituted at one or more carbon atoms by one or more -OH moieties.

[0116] As used herein, the term "isomer" means any tautomer, stereoisomer, transisomer, enantiomer, or diastereomer of any compound of the present invention. It should be understood that the compounds of the present invention may have one or more chiral centers or double bonds and thus exist in stereoisomers, such as double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). According to the present invention, the chemical structures illustrated herein and therefore the compounds of the present invention cover all corresponding stereoisomers, i.e., stereoisomeric pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers, such as racemates. Mixtures of enantiomers and stereoisomers of the compounds of the present invention can typically be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral phase gas chromatography, chiral phase high-performance liquid chromatography, recrystallization of the compound as a chiral salt complex, or recrystallization of the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained by well-known asymmetric synthetic methods from stereoisomer or enantiomer-pure intermediates, reagents, and catalysts.

[0117] As used herein, the term “stereoisomer” refers to all possible different isomers and configurations that a compound (e.g., any compound of the formula described herein) may have, especially all possible stereoisomers of the basic molecular structure.Chemical and configurational isomers, all diastereomers, enantiomers, or configurational isomers, including transisomers. Some compounds of the present invention may exist in different tautomeric forms, all of which are included within the scope of the present invention.

[0118] As used herein, the term "sulfonyl" refers to a -S(O)2- group.

[0119] As used herein, the term "thiocarbonyl" refers to a -C(S)- group.

[0120] Those skilled in the art will understand upon reading this disclosure that certain compounds described herein may be provided or used in any of a variety of forms, such as salt forms, protected forms, prodrug forms, ester forms, isomer forms (e.g., optical or structural isomers), isotopic forms, etc. In some embodiments, reference to a particular compound may refer to a particular form of the compound. In some embodiments, reference to a particular compound may refer to any form of the compound. In some embodiments, for example, a formulation of a single stereoisomer of a compound may be considered as a form of the compound different from a racemic mixture of the compound; a particular salt of a compound may be considered as a form different from another salt form of the compound; a formulation of a configurational isomer ((Z) or (E)) containing a double bond may be considered as a form different from a formulation of another configurational isomer ((E) or (Z)) containing the double bond; a formulation in which one or more atoms are isotopes different from those present in a reference formulation may be considered as a different form. Specification 31 / 123 pages 43 CN 122270438 A Detailed Description

[0121] This document provides synthetic methods and intermediates for the manufacture of Ras inhibitors or salts thereof. The methods and intermediates can be used to achieve higher yields, higher chemical purity and / or higher stereoisomer purity and lower cost for the preparation of Ras inhibitors or intermediates. Further synthetic details are provided in the examples.

[0122] The compounds described herein may be prepared using the methods described herein and / or using known organic, inorganic or enzymatic processes. Synthetic methods may employ commercially available starting materials or starting materials prepared by processes known to those skilled in the art of organic synthesis. These methods include, but are not limited to, those described in WO 2021 / 091967, WO 2022 / 235870, and WO 2023 / 060253, the disclosure of each of which is incorporated herein by reference.

[0123] In one aspect, this disclosure provides a method for isolating enantiomers of a compound of formula I.

[0124] The method of formula I comprises the steps of: a) contacting a compound of formula I with a chiral acid to form a diastereomeric salt of the compound of formula I; and b) isolating each diastereomeric isomer of the diastereomeric salt, thereby isolating the enantiomers of the compound of formula I.n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0125] In some embodiments, the nitrogen protecting group is Boc (tert-butyloxycarbonyl). In some embodiments, the chiral acid is a chiral carboxylic acid. In some embodiments, the chiral carboxylic acid is a chiral dicarboxylic acid. In some embodiments, the chiral dicarboxylic acid is tartaric acid. In some embodiments, the chiral carboxylic acid is L(+)-tartaric acid. In some embodiments, the separation step (b) includes recrystallization.

[0126] In some embodiments, the compound of formula I is further described by the compound of formula Ia: .

[0127] In some embodiments, the compound of formula I or formula Ia is compound B: .

[0128] On the other hand, this disclosure provides a barium salt of compound A: Specification 32 / 123 pages 44 CN 122270438 A.

[0129] In some embodiments, the barium salt is a 2:1 (compound A: barium) salt.

[0130] On the other hand, this disclosure provides a method for preparing a barium salt of compound A. The method comprises the following steps: a) condensing compound IIa with compound C to form compound IIb; b) cyclizing compound IIb with compound IIc to form compound IId; c) hydrolyzing compound IId to form compound IIe; d) methylating compound IIe to form compound IIf; and e) hydrolyzing compound IIf to form a barium salt of compound A: wherein R2 and R3 are each independently an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-C10 aryl, and LG is a detached group selected from the group consisting of: halogen, trifluoromethanesulfonate, methanesulfonate, and toluenesulfonate. In some embodiments, R3 is an optionally substituted C6-C10 aryl. In some embodiments, R3 is... In some embodiments, R3 is an optionally substituted C1-C6 alkyl. In some embodiments, R3 is tert-butyl. In some embodiments, R2 is an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl. In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, LG is a halogen. In some embodiments, LG is -Br.

[0131] In some embodiments of the method for preparing the barium salt of compound A, the condensation step (a) includes contacting the compound of formula IIa and compound C with a base. In some embodiments, the base is an amine base. In some embodiments, the base is a pyridine base.Pyrrolidine. In some embodiments, the condensation step (a) is performed according to the following scheme: .

[0132] In some embodiments, the condensation step (a) is performed according to the following scheme: .

[0133] In some embodiments, the condensation step (a) is performed according to the following scheme: In some embodiments, the cyclization step (b) includes contacting the compound of formula IIb and the compound of formula IIc with a base. In some embodiments, the base is a lithium base. In some embodiments, the base is LiHMDS.

[0134] In some embodiments, the cyclization step (b) is performed according to the following scheme: In some embodiments, the hydrolysis step (c) includes contacting the compound of formula IId with an acid. In some embodiments, the acid is a carboxylic acid. In some embodiments, the carboxylic acid is trifluoroacetic acid. In some embodiments, the acid is hydrochloric acid. Specification 34 / 123 Page 46 CN 122270438 A

[0135] In some embodiments, the hydrolysis step (c) is performed according to the following scheme: In some embodiments, the hydrolysis step (c) is performed according to the following scheme: In some embodiments, the hydrolysis step (c) is performed according to the following scheme: In some embodiments, the methylation step (d) includes contacting the IIe compound with MeB(OH)2. In some embodiments, the methylation step (d) further includes contacting the IIe compound with a copper catalyst. In some embodiments, the methylation step (d) is performed according to the following scheme: In some embodiments, the hydrolysis step (e) includes contacting the IIf compound with barium hydroxide.

[0136] In some embodiments, the hydrolysis step (e) is performed according to the following scheme: .

[0137] On the other hand, this disclosure provides a method for preparing a compound of formula IIf: Specification 35 / 123 pages 47 CN 122270438 A The method includes alkylating a compound of formula IIe with a methylating agent: wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl.

[0138] In some embodiments, the methylating agent includes trimethylcycloboroxane. In some embodiments, the methylating agent includes trimethylcycloboroxane and Cu(OAc)2. In some embodiments, the methylating agent includes trimethylcycloboroxane and CuOAc.

[0139] In some embodiments of the method for preparing a compound of formula IIf, the alkylation step includes contacting the compound of formula IIe with a basic salt. In some embodiments, the basic salt is potassium carbonate (K2CO3) or sodium carbonate (Na2CO3). In some embodiments, the alkylation step includes 2,2'-bipyridine. In some embodiments, the alkylation step includes di-tert-butyl peroxide (DTBP). In some embodiments, the alkylation step is performed at at least 25°C.The process is carried out at a temperature of (e.g., 25, 30, 35, or 40 °C). In some embodiments, the alkylation step is carried out in an organic solvent (e.g., acetonitrile (ACN)). In some embodiments of the method for preparing a Formula IIf compound, the alkylation step is carried out according to the following scheme: .

[0140] In some embodiments of the method for preparing a Formula IIf compound, the alkylation step is carried out according to the following scheme: .

[0141] On the other hand, this disclosure provides a method for preparing a Formula III compound or a salt thereof: .

[0142] The method of Formula III includes the following steps: a) coupling the compound of Formula IIId with the compound of Formula I to form the compound of Formula IIIe: ; and specification 36 / 123 page 48 CN 122270438 A b) deprotecting the compound of Formula IIIe to form the compound of Formula III: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; each R4 is independently an optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl; and R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0143] In some embodiments, the nitrogen protecting group is Boc (tert-butyloxycarbonyl). In some embodiments, the IIId compound is prepared by a method comprising the following steps: A1) combining the IIIa compound with the IIIb compound to form the IIIc compound; B1) reducing the IIIc compound to form the IIId compound.

[0144] Wherein Q is MgBr, MgCl, MgI, or Li.

[0145] In some embodiments, the combining step (A1) comprises contacting the IIIa compound with the IIIb compound, wherein Q is MgBr. In some embodiments, the contact is carried out in tetrahydrofuran at -78°C.

[0146] In some embodiments, the reduction step (B1) comprises contacting the IIIc compound with a ketone reductase.

[0147] In some embodiments, the IIId compound is prepared by a method comprising the following steps: A2) converting the IIIf compound to form the IIIg compound; and B2) reacting the IIIg compound (e.g., esterification) to form the IIId compound: [Specification 37 / 123, page 49, CN 122270438 A] In some embodiments, the conversion step (A2) comprises forming a diazonium salt of the IIIf compound. In some embodiments, the conversion step (A2) and the reaction (e.g., esterification) step (B2) are performed according to the following scheme:

[0148] In some embodiments, step (a) of combining the IIId compound with the I compound to form the IIIe compound includes contacting the IIId compound with trifluoromethanesulfonic anhydride, followed by contacting the IIId compound with the I compound. In some embodiments, step (a) of combining the IIId compound with the I compound to form the IIIe compound is performed according to the following scheme:

[0149] In some embodiments, the deprotection step (b) includes contacting the IIIe compound with an acid. In some embodiments, the acid is hydrochloric acid. In some embodiments, the hydrochloric acid is delivered via an ethyl acetate solution of hydrochloric acid.

[0150] On the other hand, this disclosure provides a method for preparing a compound of formula IVa: the method includes coupling a compound of formula III to a compound of formula IIf, wherein n and m are each independently 0, 1, 2, 3, 4 or 5; each R4 is independently an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; each R5 is independently an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0151] In some embodiments, R4 is benzyl, methyl or tert-butyl. In some embodiments, R5 is isopropyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R5 is cyclopentyl.

[0152] In some embodiments, coupling includes contacting the compounds of formula III and formula IIf with a base. In some embodiments, the base is 1,5,7-triazabicyclo[4,4,0]dec-5-ene (TBD). In some embodiments, coupling is carried out in an organic solvent (e.g., methyl tert-butyl ether (MTBE)). In some embodiments of the method for preparing the compound of formula IVa, coupling is carried out at a temperature of at least -10°C (e.g., -10, -5, 0, or 5°C). In some embodiments of the method for preparing the compound of formula IVa, coupling is carried out according to the following scheme: Specification 38 / 123 pages 50 CN 122270438 A.

[0153] In some embodiments of the method for preparing the compound of formula IVa, the compound of formula III is the compound of formula III-1.

[0154] In some embodiments of the method for preparing the compound of formula IVa, coupling is carried out according to the following scheme: .

[0155] In some embodiments, the compound of formula III is compound D: The compound is prepared by the following steps: a) coupling the compound of formula IIId-1 with compound B to form a compound of formula IIIe-1 having the following structure: ; and b) deprotecting the compound of formula IIIe-1 to form compound D having the following structure: .

[0156] In some embodiments, step (a) of combining formula IIId with compound B to form a compound of formula IIIe includes contacting the compound of formula IIId with trifluoromethanesulfonic anhydride, followed by contacting the compound of formula IIId with compound B.

[0157] In some embodiments, step (a) of combining formula IIId with compound B to form a compound of formula IIIe is performed according to the following scheme: Specification 39 / 123 pages 51 CN 122270438 A.

[0158] In some embodiments, the deprotection step (b) includes contacting the compound of formula IIIe with an acid. In some embodiments, the acid is hydrochloric acid. In some embodiments, the hydrochloric acid is delivered via an ethyl acetate solution of hydrochloric acid.

[0159] On the other hand, this disclosure provides a method for preparing a compound of formula IV or a salt thereof: .

[0160] The method includes the following steps: a) coupling compound III with a barium salt of compound A to form compound IVa; and b) hydrolyzing compound IVa to form compound IV: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-.

[0161] In some embodiments, the coupling step (a) includes contacting compound III and barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH). In some embodiments, the coupling step (a) further includes contacting the compound of formula III and the barium salt of compound A with NMI. In some embodiments, the coupling step (a) is carried out according to the following scheme: Specification 40 / 123 pages 52 CN 122270438 A.

[0162] In some embodiments, the hydrolysis step (b) includes contacting the compound of formula IVa with an acid or base. In some embodiments, the hydrolysis step (b) includes hydrogenating the compound of formula IVa. In some embodiments, hydrogenation further includes contacting the compound of formula IVa with a palladium catalyst.

[0163] In some embodiments, the hydrolysis step (b) is carried out according to the following scheme: In some embodiments of any methods and compounds described herein, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0164] In some embodiments of any methods and compounds described herein, each of X1, X2 and X3 isCH2. In some embodiments, PG is. In some embodiments, LG is a halogen. In some embodiments, LG is Br.

[0165] In some embodiments of any of the methods and compounds described herein, R2 is an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl. In some embodiments, R2 is benzyl, methyl, or tert-butyl. In some embodiments, R3 is an optionally substituted C6-C10 aryl. In some embodiments, R3 is. In some embodiments, R3 is an optionally substituted C1-C6 alkyl. In some embodiments, R3 is. In some embodiments, R4 is an optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl. In some embodiments, R4 is benzyl, methyl, or tert-butyl. In some embodiments, R5 is an optionally substituted C1-C6 alkyl or optionally substituted 3- to 6-membered cycloalkyl. In some embodiments, R5 is an optionally substituted C1-C6 alkyl. In some embodiments, R5 is isopropyl. In some embodiments, R5 is an optionally substituted 3- to 6-membered cycloalkyl group. In some embodiments, R5 is cyclopentyl.

[0166] In some embodiments, the method includes the preparation of compound E: Specification 41 / 123 pages 53 CN 122270438 A and the method includes the steps of: a) coupling compound D with a barium salt of compound A to form a compound of formula IVa having the following structure: ; and b) hydrolyzing compound IVa to form compound E having the following structure: .

[0167] In some embodiments, the coupling step (a) includes contacting compound D and the barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH). In some embodiments, the coupling step (a) further includes contacting compound D and the barium salt of compound A with NMI. In some embodiments, the coupling step (a) is carried out according to the following scheme: .

[0168] In some embodiments, the hydrolysis step (b) includes contacting the IVa compound with an acid or a base. In some embodiments, the hydrolysis step (b) includes hydrogenating the IVa compound. In some embodiments, hydrogenation further includes contacting the IVa compound with a palladium catalyst. In some embodiments, the hydrolysis step (b) is performed according to the following scheme: On the other hand, this disclosure provides a method for preparing a VIII compound or a salt thereof: Specification 42 / 123 pages 54 CN 122270438 A The method includes the following steps: a) esterifying and N-alkylating the VIIIa compound to form a VIIIb compound; b) reducing the VIIIb compound to form a VIIIc compound in the form of a mixture of transisomers; and c)A mixture of isolated transisomers of formula VIIIc is obtained to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0169] In some embodiments, the esterification and N-alkylation step (a) comprises contacting the compound of formula VIIIa with an alkylating agent. In some embodiments, the alkylating agent is R13-LG, wherein R13 is an optionally substituted C1-C6 alkyl group and LG is a detachable group. In some embodiments, the alkylating agent is 2,2,2-trifluoroethyl trifluoromethanesulfonate.

[0170] In some embodiments, the esterification and N-alkylation step (a) is carried out according to the following scheme: In some embodiments, the reduction step (b) comprises contacting the compound of formula VIIIb with a reducing agent. In some embodiments, the reducing agent is a borohydride. In some embodiments, the borohydride is lithium borohydride. In some embodiments, the reduction step (b) is performed according to the following scheme: Specification 43 / 123 Page 55 CN 122270438 A In some embodiments, the compound of formula VIII is compound F: and the method includes the following steps: a) esterifying and N-alkylating the compound of formula VIIIa to form a compound of formula VIIIb having the following structure: b) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers having the following structure: and c) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII.

[0171] In some embodiments, the esterification and N-alkylation step (a) includes contacting the compound of formula VIIIa with an alkylating agent. In some embodiments, the alkylating agent is R13-LG, wherein R13 is an optionally substituted C1-C6 alkyl group and LG is a detachable group. In some embodiments, the alkylating agent is 2,2,2-trifluoroethyl trifluoromethanesulfonate. In some embodiments, the esterification and N-alkylation steps (a) are performed according to the following scheme: Specification 44 / 123 pages 56 CN 122270438 A In some embodiments, the reduction step (b) includes contacting the compound of formula VIIIb with a reducing agent. In some embodiments, the reducing agent is a borohydride. In some embodiments, the borohydride is lithium borohydride.

[0172] In some embodiments, the reduction step (b) is performed according to the following scheme: On the other hand, this disclosure provides a method for preparing a compound of formula VIII or a salt thereof: The method includes the following steps: a) reacting the compound of formula VIIIa (e.g., esterification) to form a compound of formula VIIId: ; b) N-alkylating the compound of formula VIIId to form a compound of formula VIIIb: ; Specification 45 / 123 pages 57 CN122270438 A c) Reducing the VIIIb compound to form a mixture of VIIIc compounds in the form of transisomers: ; and d) Separating the mixture of transisomers of VIIIc to obtain a stereochemically pure VIII compound; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0173] In some embodiments, step (a) includes contacting the VIIIa compound with R13OH, wherein R13 is an optionally substituted C1-C6 alkyl group. In some embodiments, the alcohol is methanol. In some embodiments, step (a) further includes contacting the VIIIa compound with an acid. In some embodiments, the acid is sulfuric acid. In some embodiments, step (a) is carried out according to the following scheme: In some embodiments, the N-alkylation step (b) includes contacting the VIIId compound with an alkylating agent. In some embodiments, the alkylating agent is R13-LG, wherein R13 is an optionally substituted C1-C6 alkyl group and LG is a release group. In some embodiments, the alkylating agent is 2,2,2-trifluoroethyl trifluoromethanesulfonate. In some embodiments, the N-alkylation step (b) is performed according to the following scheme: In some embodiments, the reduction step (c) includes contacting a compound of formula VIIIb with a reducing agent. In some embodiments, the reducing agent is a borohydride. In some embodiments, the borohydride is lithium borohydride.

[0174] In some embodiments, the reduction step (c) is performed according to the following scheme: Specification 46 / 123 page 58 CN 122270438 A In some embodiments, the compound of formula VIII is compound G: and the method includes the following steps: a) reacting the compound of formula VIIIa (e.g., esterification) to form a compound of formula VIIId having the following structure: b) N-alkylating the compound of formula VIIId to form a compound of formula VIIIb having the following structure: ; c) reducing the compound of formula VIIIb to form a compound G in the form of a mixture of transisomers having the following structure: In some embodiments, step (a) includes contacting the compound of formula VIIIa with R13OH, wherein R13 is an optionally substituted C1-C6 alkyl. In some embodiments, the alcohol is methanol. In some embodiments, step (a) further includes contacting the compound of formula VIIIa with an acid. In some embodiments, the acid is sulfuric acid. Specification 47 / 123 page 59 CN 122270438 A In some embodiments, step (a) is performed according to the following scheme: In some embodiments, the N-alkylation step (b) includes contacting the compound of formula VIIId with an alkylating agent.In some embodiments, the alkylating agent is R13-LG, wherein R13 is an optionally substituted C1-C6 alkyl group and LG is a detachable group. In some embodiments, the alkylating agent is 2,2,2-trifluoroethyl trifluoromethanesulfonate. In some embodiments, the N-alkylation step (b) is performed according to the following scheme: In some embodiments, the reduction step (c) includes contacting the compound of formula VIIIb with a reducing agent. In some embodiments, the reducing agent is a borohydride. In some embodiments, the borohydride is lithium borohydride. In some embodiments, the reduction step (c) is performed according to the following scheme: In some embodiments, the mixture of transisomers is separated by column chromatography or chemical resolution. In some embodiments, chemical resolution includes contacting the compound of formula VIIIc with an acid. In some embodiments, the acid is methanesulfonic acid. In some embodiments, the acid is camphorsulfonic acid (e.g., D-camphor-10-sulfonic acid).

[0175] On the other hand, this disclosure provides a method for preparing a compound of formula VIIIc: Specification 48 / 123 pages 60 CN 122270438 A The method comprises N-alkylating a compound of formula VIIIe with a compound having the structure R13-LG1: wherein each R12 is independently a halogen; each R13 is independently an optionally substituted C1-C6 alkyl group; and LG1 is a detachable group.

[0176] In some embodiments, each R12 is Br. In some embodiments, each R13 is -CH2CF3. In some embodiments, LG1 is a halogen, a trifluoromethanesulfonate group, a methanesulfonate group, or a toluenesulfonate group. In some embodiments, R13-LG is 2,2,2-trifluoroethyl trifluoromethanesulfonate. In some embodiments, the method comprises contacting the compound of formula VIIIe and R13-LG1 with a base. In some embodiments, the base is tripotassium phosphate (K3PO4). In some embodiments, the method is carried out at a temperature of at least 10°C (e.g., 10, 15, 20, or 25°C). In some embodiments, the method produces a mixture of transisomers of the formula VIIIc compound. In some embodiments, N-alkylation is carried out according to the following scheme:

[0177] In some embodiments of the method for preparing the formula VIIIc compound, the formula VIIIe compound is compound K. In some embodiments, the formula VIIIc compound is compound G. In some embodiments, N-alkylation is carried out according to the following scheme: Specification 49 / 123 pages 61 CN 122270438 A.

[0178] On the other hand, this disclosure provides a method for preparing the formula VIIIe compound: The method includes reducing the formula VIIIe compound with a reducing agent: wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0179] In some embodiments, each R12 is Br. In some embodiments, each R13 is -CH2CF3. In some embodiments, the reducing agent is NaBH4. In some embodiments, the method includes contacting the compound of formula VIIIf with calcium chloride (CaCl2). In some embodiments, the reduction is carried out at a temperature of at least 20°C (e.g., 20, 25, or 30°C). In some embodiments, the reduction is carried out according to the following scheme: .

[0180] In some embodiments, the reduction is carried out according to the following scheme: Specification 50 / 123 pages 62 CN 122270438 A.

[0181] On the other hand, this disclosure provides a method for preparing a compound of formula IX or a salt thereof: the method includes the steps of: a) borate a compound of formula VIII to form a compound of formula X: ; and b) couple a compound of formula X to a compound of formula XI to form a compound of formula XII: ; wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; and R14 is an optionally substituted 3- to 6-membered cycloalkyl group.

[0182] In some embodiments, the borate step (a) includes contacting a compound of formula VIII with a borater. In some embodiments, the borater is bis(pinacol)diboron (B2Pin2). In some embodiments, the borate step (a) further includes contacting a compound of formula VIII with 4,4'-di-tert-butyl-2,2'-dipyridyl (dtbpy). In some embodiments, page 51 / 123 of document 63 CN 122270438 A, the borate step (a) further includes contacting the compound of formula VIII with an iridium catalyst. In some embodiments, the iridium catalyst is [Ir(OMe)(COD)]2.

[0183] In some embodiments, the borate step (a) is performed according to the following scheme: In some embodiments, the coupling step (b) includes contacting the compounds of formula X and formula XI with a copper catalyst. In some embodiments, the copper catalyst is Cu(OAc)2. In some embodiments, the coupling step (b) includes contacting the compounds of formula X and formula XI with a base. In some embodiments, the base is an amine base. In some embodiments, the base is triethylamine or tetramethylpiperidine. In some embodiments, the coupling step (b) is performed according to the following scheme: In some embodiments, the compound of formula IX is compound H: The method includes the following steps: a) borate the compound of formula VIII to form the compound of formula X having the following structure: Specification 52 / 123 pages 64 CN 122270438 A; and b) couple the compound of formula X with the compound of formula XI to form the compound H having the following structure: .

[0184] In some embodiments, the borate step (a) includes contacting compound VIII with a borate agent. In some embodiments, the borate agent is bis(pinacol)diboron (B2Pin2). In some embodiments, the borate step (a) further includes contacting compound VIII with 4,4'-di-tert-butyl-2,2'-dipyridyl (dtbpy). In some embodiments, the borate step (a) further includes contacting compound VIII with an iridium catalyst. In some embodiments, the iridium catalyst is [Ir(OMe)(COD)]2. In some embodiments, the borate step (a) is performed according to the following scheme: In some embodiments, the coupling step (b) includes contacting compounds X and XI with a copper catalyst. In some embodiments, the copper catalyst is Cu(OAc)2. In some embodiments, the coupling step (b) includes contacting compounds X and XI with a base. In some embodiments, the base is an amine base. In some embodiments, the base is triethylamine or tetramethylpiperidine. In some embodiments, the coupling step (b) is carried out according to the following scheme: Specification 53 / 123 page 65 CN 122270438 A In some embodiments, the compound of formula X or a salt thereof: is further prepared by the following steps: a) borate the compound of formula VIII to form the compound of formula XIII: ; and b) hydrolyze and borate the compound of formula XIII in one step to form formula X: ; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group.

[0185] In some embodiments, the borate step (a) includes contacting the compound of formula VIII with a borater. In some embodiments, the borater is pinacolborane (HBPin). In some embodiments, the borate step (a) is carried out according to the following scheme: Specification 54 / 123 page 66 CN 122270438 A In some embodiments, the hydrolysis and borate step (b) includes contacting the compound of formula XIII with a borater. In some embodiments, the borate agent is bis(pinacol)diboron (B2Pin2). In some embodiments, the hydrolysis and borate step (b) further includes contacting the compound of formula XIII with 4,4'-di-tert-butyl-2,2'-dipyridyl (dtbpy). In some embodiments, the hydrolysis and borate step (b) further includes contacting the compound of formula XIII with an iridium catalyst. In some embodiments, the iridium catalyst is [Ir(OMe)(COD)]2. In some embodiments, the hydrolysis and borate step (b) is performed according to the following scheme: On the other hand, this disclosure provides a method for preparing a compound of formula XIV or a salt thereof: The method includes the following steps: a)a) Reacting a compound of formula XV with a compound of formula XII (e.g., esterification) to form a compound of formula XVI: (See specification 55 / 123, page 67, CN 122270438 A); b) Removing (e.g., deprotecting) the PGa of a compound of formula XVI to form a compound of formula XVII: ; c) Cycling a compound of formula XVII with a transition metal catalyst to form a compound of formula XVII: ; d) Removing (e.g., deprotecting) the PGa of a compound of formula XVIII to form a compound of formula XIX: ; and e) Coupled with a compound of formula XIX to form a compound of formula IV: (See specification 56 / 123, page 68, CN 122270438 A); wherein n and m are each independently 0, 1, 2, 3, 4, or 5; X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-; R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; PGa is an acid-instability protecting group; and PGb is an acid-stability protecting group.

[0186] In some embodiments, step (a) includes contacting the compounds of formula XV and XII with a carbodiimide coupling agent, an anti-racemic agent, and a base. In some embodiments, the carbodiimide coupling agent is EDCl, the anti-racemic agent is HOBt, and the base is N,N-diisopropylethylamine.

[0187] In some embodiments, step (a) is performed according to the following scheme: In some embodiments, the deprotection step (b) includes contacting the compound of formula XVI with an acid. In some embodiments, the acid is trifluoroacetic acid. In some embodiments, the deprotection step (b) is performed according to the following scheme: In some embodiments, the cyclization step (c) includes contacting the compound of formula XVII with a transition metal catalyst, a phosphine ligand, and a base. In some embodiments, the transition metal catalyst includes palladium, platinum, nickel, copper, and iron catalysts. In some embodiments, the transition metal catalyst is a palladium catalyst. In some embodiments, the palladium catalyst is P(tBu)3PdG3 or Pd2(dba)3 as described on page 57 / 123 of the specification, CN 122270438 A. In some embodiments, the phosphine ligand is Q-Phos or [(tBu)3PH]BF4. In some embodiments, the base is K3PO4. In some embodiments, the cyclization step (c) is performed according to the following scheme: In some embodiments, the cyclization step (c) is performed according to the following scheme: In some embodiments, the cyclization step (c) further includes contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII.

[0188] In some embodiments, the deprotection step (d) includes contacting the compound of formula XVIII with a palladium catalyst and hydrogen in an organic solvent. In some embodiments, the palladium catalyst is Pd / C. In some embodiments, the organic solvent is methyl tert-butyl ether. In some embodiments, the deprotection step (d) is performed according to the following scheme: In some embodiments, the deprotection step (d) further includes washing the compound of formula XIX with N-acetylcysteine.

[0189] In some embodiments, the deprotection step (e) includes contacting the compound of formula XIX and the compound of formula IV with a coupling agent, an anti-racemic agent, and a base. In some embodiments, the coupling agent is PyBOP or EDCl. In some embodiments, the anti-racemic agent is selected from the group consisting of Oxyma, HOBt, or HOPO. In some embodiments, the base is N,N-diisopropylethylamine.

[0190] In some embodiments, the coupling step (e) is performed according to the following scheme: In some embodiments, the coupling step (e) is performed according to the following scheme: In some embodiments, the coupling step (e) is performed according to the following scheme: In some embodiments, the method further includes a step of purifying the compound of formula XIV. In some embodiments, purification includes recrystallizing the compound of formula XIV. In some embodiments, recrystallization includes adding a first solvent, followed by adding a second solvent. In some embodiments, the first solvent is dioxane or 2-methyltetrahydrofuran. In some embodiments, the second solvent is diisopropyl ether or isopropanol. In some embodiments, recrystallization further includes adding a third solvent. In some embodiments, the third solvent is heptane. In some embodiments, R12 is -Br. In some embodiments, R13 is -CH2CF3. In some embodiments, PGa is Boc. In some embodiments, PGb is CBz.

[0191] In some embodiments, the compound of formula XIV is compound 1: (Specification page 59 / 123, 71 CN 122270438 A) Compound 1, and the method includes the following steps: a) reacting the compound of formula XV with the compound of formula XII (e.g., esterification) to form the compound of formula XVI having the following structure: ; b) deprotecting the Boc of the compound of formula XVI to form the compound of formula XVII having the following structure: ; c) cyclizing the compound of formula XVII with a transition metal catalyst to form the compound of formula XVII having the following structure: (Specification page 60 / 123, 72 CN 122270438 A) ; d) removing (e.g., deprotecting) the PGb of the compound of formula XVIII to form the compound of formula XIX having the following structure: ; and e)The compound of formula XIX is coupled with the compound of formula IV to form compound 1 having the following structure: .

[0192] In some embodiments, step (a) includes contacting the compounds of formula XV and XII with a carbodiimide coupling agent, an anti-racemic agent, and a base. In some embodiments, the carbodiimide coupling agent is EDCl, the anti-racemic agent is HOBt, and the base is N,N-diisopropylethylamine. In some embodiments, step (a) is performed according to the following scheme: Specification 61 / 123 pages 73 CN 122270438 A In some embodiments, the deprotection step (b) includes contacting the compound of formula XVI with an acid. In some embodiments, the acid is trifluoroacetic acid. In some embodiments, the deprotection step (b) is performed according to the following scheme: In some embodiments, the cyclization step (c) includes contacting the compound of formula XVII with a transition metal catalyst, a phosphine ligand, and a base. In some embodiments, the transition metal catalyst includes palladium, platinum, nickel, copper, and iron catalysts. In some embodiments, the transition metal catalyst is a palladium catalyst. In some embodiments, the palladium catalyst is P(tBu)3PdG3 or Pd2(dba)3. In some embodiments, the phosphine ligand is Q-Phos or [(tBu)3PH]BF4. In some embodiments, the base is K3PO4.

[0193] In some embodiments, the cyclization step (c) is performed according to the following scheme: In some embodiments, the cyclization step (c) is performed according to the following scheme: Specification 62 / 123 pages 74 CN 122270438 A In some embodiments, the cyclization step (c) further includes contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII.

[0194] In some embodiments, the deprotection step (d) includes contacting the compound of formula XVIII with a palladium catalyst and hydrogen in an organic solvent. In some embodiments, the palladium catalyst is Pd / C. In some embodiments, the organic solvent is methyl tert-butyl ether. In some embodiments, the deprotection step (d) is performed according to the following scheme: In some embodiments, the deprotection step (d) further includes washing the compound of formula XIX with N-acetylcysteine.

[0195] In some embodiments, the deprotection step (e) includes contacting the compound of formula XIX and the compound of formula IV with a coupling agent, an anti-racemic agent, and a base. In some embodiments, the coupling agent is PyBOP or EDCl. In some embodiments, the anti-racemic agent is selected from the group consisting of Oxyma, HOBt, or HOPO. In some embodiments, the base is N,N-diisopropylethylamine.

[0196] In some embodiments, the coupling step (e) is performed according to the following scheme: Specification 63 / 123Page 75 CN 122270438 A In some embodiments, the coupling step (e) is performed according to the following scheme: In some embodiments, the coupling step (e) is performed according to the following scheme: In some embodiments, the method further includes a step of purifying compound 1. In some embodiments, purification includes recrystallizing compound 1. In some embodiments, recrystallization includes adding a first solvent, followed by adding a second solvent. In some embodiments, the first solvent is dioxane or 2-methyltetrahydrofuran. In some embodiments, the second solvent is diisopropyl ether or isopropanol. In some embodiments, recrystallization further includes adding a third solvent. In some embodiments, the third solvent is heptane.

[0197] Compounds prepared by the methods described herein containing aziridine compounds can be used as crosslinking groups.

[0198] For example, compounds of formula IV contain an aziridine moiety that can act as an electrophile.

[0199] Those skilled in the art will be familiar with nucleophiles that can react with aziridine electrophiles. For example, nucleophilic amino acids (e.g., cysteine, aspartic acid, glutamic acid, tyrosine, arginine, histidine, or lysine) can react with aziridine in the compound of formula IV.

[0200] The compound of formula IV can be combined with a monovalent organic moiety. Organic moieties are well known to those skilled in the art. The monovalent organic moiety can be, for example, a small molecule (e.g., a macrocyclic small molecule), a polymer, a nucleic acid (e.g., a DNA or RNA oligonucleotide), a polypeptide, an oligosaccharide, an organometallic compound, or a protein (such as a mutant protein). The organic moiety can be combined with the compound of formula IV as disclosed herein in a variety of ways, and methods for attaching synthetic intermediates of formula IV as described herein to the monovalent organic moiety are well known to those skilled in the art. Exemplary methods are disclosed in WO 2021 / 091967, WO 2022 / 235870, and WO 2023 / 060253, the disclosures of each of which are incorporated herein by reference.

[0201] Methods for combining a carboxylic acid moiety with other compounds will be well known to those skilled in the art. For example, in some embodiments described on page 64 / 123 of CN 122270438 A, the compound of formula IV can be reacted with an alcohol in an esterification reaction. In some embodiments, the compound of formula IV can be reacted with an amine in a peptide coupling reaction. Peptide coupling reactions will be familiar to those skilled in the art. Non-limiting examples of peptide coupling agents that can be used to react the carboxylic acid moiety of formula IV with an amine include carbodiimide coupling agents (e.g., DCC, DIC, and EDC), ureonium coupling agents (e.g., COMU, HATU, HBTU, HCTU, TATU, TOTU, TBTU), carbonyl diimidazole (CDI), and phosphonium coupling agents.(e.g., BOP, PyBOP, BOP-Cl, PyAOP, PyBOP). In some embodiments, 1-hydroxybenzotriazole (HOBt) may be added to the peptide coupling reaction.

[0202] In some embodiments, the carboxylic acid may first be converted to an ester, which may react directly with a nucleophile.

[0203] For example, a compound of formula IV may be converted to a compound of formula V: wherein R6 is, , , , , , , or.

[0204] In some embodiments, a compound of formula IV or V is reacted with an amine of formula VI or a salt thereof: Formula VI Wherein the dashed line represents zero, one, two, three or four non-adjacent double bonds; A is -N(H or CH3)C(O)-(CH2)-, wherein the amino nitrogen is bonded to the carbon atom of -CH(R10)-; optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heteroalkylene, optionally substituted 6-membered aryl or optionally substituted 5- to 10-membered heteroaryl; Specification 65 / 123 pages 77 CN 122270438 A X1 is optionally substituted C1-C2 alkylene, NR, O or S(O)n; X2 is O or NH; X3 is N or CH; n is 0, 1 or 2; R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R' or S(O)2N(R')2; each R' is independently H or an optionally substituted C1-C4 alkyl; Y1 is C, CH or N; Y2, Y3, Y4 and Y7 are independently C or N; Y5 is CRx, CH2 or N; Y6 is C(O), CRz, CH2 or N; Rx is hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocyclic alkyl; Ry is hydrogen, halogen, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 alkoxy, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered heterocyclic alkyl; R1 is a cyano group, optionally substituted C1-C6 alkyl group, optionally substituted C1-C6 heteroalkyl group, optionally substituted 3- to 6-membered cycloalkyl group, optionally substituted 3- to 6-membered cycloalkenyl group, optionally substituted 3- to 6-membered heterocycloalkyl group, optionally substituted 6- to 10-membered aryl group, or optionally substituted 5- to 10-membered heteroaryl group, or R1 and R2 combined with the atoms they are attached to form optionally substituted 3- to 14-membered heterocycloalkyl groups; R2 is absent, and is hydrogen, optionally substituted C1-C6 alkyl group, optionally substituted C2-C6 alkenyl group, optionally substituted C2-C6 ynyl group, optionally substituted 3- to 6-membered cycloalkyl group, optionally substituted 3- to 7-membered heterocycloalkyl group, optionally substituted 6-membered aryl group, optionally substituted 5- to 10-membered heteroaryl group, or optionally substituted 3- to 6-membered cycloalkyl group.The substituted 5- or 6-membered heteroaryl group; R3 is absent, or R2 and R3, together with the atoms to which they are attached, form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl; R4 is absent, and is hydrogen, halogen, cyano, or a methyl group optionally substituted with 1 to 3 halogens; R5 is hydrogen, an optionally substituted C1-C4 alkyl group, cyano, hydroxyl, or C1-C4 alkoxy, cyclopropyl, or cyclobutyl; R6 is hydrogen or methyl; R7 is hydrogen, halogen, or an optionally substituted C1-C3 alkyl group, or R6 and R7, together with the carbon atoms to which they are attached, form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R8 is hydrogen, halogen, hydroxyl, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocyclic alkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7 and R8 combined with the carbon atom to which they are attached to form C=CR7'R8'; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocyclic alkyl; R7a and R8a are independently hydrogen, halogen, optionally substituted C1-C3 alkyl, or combined with the carbon atom to which they are attached to form a carbonyl group; R7' is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R8' is hydrogen, halogen, hydroxyl, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R7' and R8' combined with the carbon atoms they are attached to form optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 7-membered heterocycloalkyl; R9 is hydrogen, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; or R9 and L combined with the atoms they are attached to form optionally substituted 3- to 14-membered heterocycloalkyl; R9' is hydrogen or optionally substituted C1-C6 alkyl; R10 is hydrogen, halogroup, hydroxyl group, C1-C3 alkoxy group, or C1-C3 alkyl; R10a is hydrogen or halogroup; R11 is hydrogen or C1-C3 alkyl; and R34 is hydrogen or C1-C3 alkyl (e.g., methyl).

[0205] In some embodiments, the compound of formula IV or V is reacted with an amine of formula VII or a salt thereof: Formula VIIWherein A is an optionally substituted 3- to 6-membered heterocyclic alkylene, optionally substituted 3- to 6-membered heterocyclic alkylene, optionally substituted 6-membered arylene, or optionally substituted 5- to 10-membered heterocyclic arylene; R1 is hydrogen or optionally substituted 3- to 10-membered heterocyclic alkylene; and R2 is an optionally substituted C1-C6 alkylene, optionally substituted C3- to 6-cycloalkyl, or optionally substituted C6- to 10 arylene.

[0206] Examples This disclosure is further illustrated by the following examples and synthetic examples, which should not be construed as limiting the scope or spirit of this disclosure to the specific procedures described herein. It should be understood that examples are provided to illustrate certain embodiments and are not intended to limit the scope of this disclosure thereby. It should be further understood that various other embodiments, modifications and equivalents thereof may be conceived by those skilled in the art without departing from the spirit of this disclosure or the scope of the appended claims.

[0207] All variables described in the following examples have the same definitions as defined in the above summary of the invention.

[0208] Example 1 - Synthetic compound I. Instructions for Use, Page 67 / 123, 79, CN 122270438 A

[0209] : A racemic mixture of compound I (8.40 mol, 1.0 equiv), EtOH (9.6 V), L(+)-tartaric acid (0.5 equiv), and H2O (0.35 V) were charged into a reactor. The reaction mixture was purged three times with N2 gas and then stirred at 80°C for 3 h. The reactants were then slowly cooled to 25°C over 12 h. The reaction mixture was stirred at 20–25°C for 4 h. The slurry was filtered, and the filter cake was washed with EtOH (1 V). The wet filter cake was charged into the reactor along with EtOH (3 V) and H2O (0.105 V). The mixture was purged three times with N2 gas and then stirred at 80°C for 3 h. The reaction mixture was then slowly cooled to 25°C over 12 h. The reaction mixture was stirred at 20–25°C for 4 h. The slurry was filtered, and the filter cake was washed with EtOH (1 V) and MTBE (1 V), respectively. The filter cake was dried under reduced pressure (40 °C, 50-80 mbar) to give a grayish-white solid tartrate of compound I (100% a / a purity, 93.3% ee, 39% yield). Table 1 summarizes the HPLC method used for this synthesis.

[0210] Table 1: HPLC method used for Example 1 Chiral HPLC method:

[0211] Chiral HPLC method: Specification 68 / 123 pages 80 CN 122270438 A

[0212] Example 2 - Synthesis procedure of (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylate and (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylic acid (barium salt of compound A) Part 1Synthesis of (R,E)-N-(cyclopropylmethylene)-4-methylbenzenesulfinamide: Cyclopropane formaldehyde (5.78 kg, 82.46 mol, 2.0 equiv), pyrrolidine (14.66 g, 0.41 mol, 0.5 mol%), and DCM (128.00 L, 20 V) were charged into a reactor. The reaction mixture was stirred at 20–25 °C for 30 min. A cyclopropane formaldehyde solution (64.00 L, 41.2 mol, 10 V) was transferred to a jacketed reactor equipped with a distillation apparatus, followed by the addition of starting material (6.4 kg, 1.0 equiv). The reaction mixture was distilled at 40–45 °C to remove H₂O. When the reaction solution was less than 7 V, another portion of cyclopropane formaldehyde solution (19.20 L, 3 V) was added. The cyclopropane formaldehyde solution was completely added over 53 h. The reaction was monitored by HPLC (IPC ≤ 1% a / a). After completion, the reactants were cooled to 20–25 °C and the reaction solution was filtered. The filtrate was concentrated to 4 V under reduced pressure (35–40 °C) and loaded with n-heptane (10.00 L, 1.5 V). The mixture was concentrated to 4 V under reduced pressure (45–49 °C) and loaded with n-heptane (10.00 L, 1.5 V). The process was repeated 4 times until the solvent was replaced with heptane. The mixture was stirred at 15–20 °C for 14 h. The slurry was filtered and the wet filter cake was dried under reduced pressure (35–40 °C, 100–200 mbar) for 16 h. A product in the form of a pale yellow solid was obtained (95.8% a / a, 93.7% w / w, 84% yield).

[0213] LCMS (ESI+): C11H13NOS [M+H]+ Calculated value: 208.1; Experimental value: 208.1. 1H NMR (400 MHz, DMSO-d 6) δ 7.58-7.50 (m, 3H), 7.38 (d, J=8.0 Hz, 2H), 2.37 (s, 3H), 2.00-1.87 (m, 1H), 1.13‑0.92 (m, 4H).

[0215] Part 2 - Synthesis of (2R,3R)-3-cyclopropyl-1-((R)-p-tolylsulfinyl)aziridine-2-carboxylate, specification page 69 / 123, CN 122270438 A. Under N2 protection, (R,E)-N-(cyclopropylmethylene)-4-methylbenzenesulfinamide (1.0 kg, 4.82 mol, 1.0 equiv), benzyl bromoacetate (1.44 kg, 6.29 mol, 1.3 equiv), and THF (10.00 L, 10The reaction mixture was cooled to -50°C to -40°C and stirred for 30 min. LiHMDS (1.0 M in THF, 4.82 mol, 4.82 L, 1.0 equiv) was added dropwise to the reaction mixture over 2.5 h at -45 ± 5°C. After completion, the reaction was quenched by pouring the reaction mixture into ice water (20.00 L, 20 V). MTBE (10.00 L, 10 V) was added, and the mixture was stirred at 10–20°C for 10 min. The mixture was allowed to stand for 30 min, after which the organic phase was separated. The aqueous phase was extracted with MTBE (10.00 L, 10 V). The combined organic phases were washed with an aqueous NaCl solution (10 wt%, 10.00 L, 10 V). The organic phase was concentrated under reduced pressure (25–35°C, 50–100 mbar). The residue was dissolved in MTBE (10.00 L, 10 V) and loaded with DABCO (81.10 g, 0.72 mol, 0.15 equiv). The mixture was stirred for 9 h and sampled for HPLC analysis (IPC: benzyl bromoacetate = 0% a / a). The combined organic phases were washed with NaCl aqueous solution (10 wt%, 10.00 L, 10 V). The organic phases were concentrated under reduced pressure (25–35 °C, 50–100 mbar) to give a crude product in the form of a brown oil (80.4% a / a purity).

[0216] The crude product combined with different batches (7.5 kg, 1.0 equiv) and isopropyl ether (5.25 L, 0.7 V) was added to the reactor. The mixture was stirred at 20–25 °C for 30 min and then cooled to 0 ± 5 °C. Product seed crystals (1.0 g) were added. The mixture was cooled to -15°C to -20°C and stirred at this temperature for 2 h. The slurry was filtered, and the filter cake was washed with isopropyl ether (pre-cooled to -20°C, 0.75 L, 0.1 V). The wet filter cake was dried to give a pale yellow solid (3.1 kg, 89.1% a / a purity). The solid was dissolved in isopropyl ether (2.70 L, 0.9 V). The solution was cooled to -15°C to -20°C and stirred at this temperature for 1 h. The slurry was filtered, and the filter cake was washed with isopropyl ether (pre-cooled to -20°C, 0.60 L, 0.2 V). The wet filter cake was dried to give a pale yellow solid of the product (2.48 kg, 98.3% a / a purity).

[0217] LCMS (ESI+): C20H21NO3S [M+H]+ Calculated value: 356.1; Experimental value: 356.3.

[0218] 1H NMR (400 MHz, DMSO-d 6) δ 7.63-7.56 (m, 2H), 7.40-7.29 (m, 5H),7.19‑7.10 (m, 2H) , 5.14‑5.02 (m, 2H) , 3.15 (d ,J= 7.1 Hz, 1H) , 2.38 (s, 3H) , 2.35 (d ,J= 7.5 Hz, 1H) , 1.05‑ 0.92 (m, 1H), 0.61-0.44 (m, 3H), 0.44-0.34 (m, 1H).

[0219] Partial 3-Synthesis of (2R,3R)-3-cyclopropylaziridine-2-carboxylate: (2R,3R)-3-cyclopropyl-1-((R)-p-tolylsulfinyl)aziridine-2-carboxylate (3.00 kg, 8.44 mol, 1.0 equiv), acetone (30.00 L, 10 V), and H2O (15.00 L, 5 V) were charged into the reactor. The reaction mixture was cooled to -10°C to -5°C. TFA (4.81 kg, 42.20 mol, 5.0 equiv) was added dropwise to the reactants over 90 min at -15°C to -5°C. The reaction mixture was stirred at -10°C to 0°C for 3 h.

[0220] The reaction was quenched with an aqueous ammonia solution (25% w / w, 1 V, 7.0 equiv). Add MTBE (45.00 L, 15 V) and separate the phases. Extract the aqueous phase with MTBE (1 x 30.00 L, 1 x 10 V; 3 x 15.00 L, 3 x 5 V). Wash the combined organic phases with NaCl aqueous solution (13% w / w, 2 x 30.00 L, 2 x 10 V). Concentrate the organic phase and exchange it with MTBE (10.00 L x 3) to remove acetone. Wash the MTBE solution with water (30.00 L, 10 V). Concentrate the organic phase to dryness to give an oily substance (25–35 °C, 50–100 mbar). Dissolve the residue in isopropyl ether (2.50 L, 0.42 V) and add n-heptane (2.00 L, 0.33 V) to the solution. The solution was cooled to 10°C and product seed crystals (20 g) were added. The mixture was stirred for 1 h and then cooled to 5–10°C. Heptane (2.00 L, 0.33 V) was added to the solution and the mixture was stirred at 5–10°C for 2 h. The slurry was filtered at 5–10°C and the filter cake was washed with isopropyl ether / heptane (1:5 v / v, 0.2 V). The filter cake was dried under reduced pressure (100–200 mbar, 25°C) to give the product (1.50 kg, 92.1% a / a, 89.6% w / w analysis, 100% ee, 99.4% de).

[0221] LCMS (ESI+):C13H15NO2[M+H]+ Calculated value: 218.1; Experimental value: 218.2.

[0222] 1H NMR (400 MHz, CDCl3) δ 7.43-7.31 (m, 5H), 5.32-5.20 (m, 2H), 2.73 (d, J= 6.0 Hz, 1H), 1.68 (dd, J= 8.2, 6.0 Hz, 1H), 0.83-0.70 (m, 1H), 0.65-0.56 (m, 1H), 0.54-0.40 (m, 2H), 0.36-0.25 (m, 1H).

[0223] Partial 4-synthesis of (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylate: (2R,3R)-3-cyclopropylaziridine-2-carboxylate (10.90 kg, 50.17 mol, 1.0 equiv), DMF (109.00 L, 10 V), MeB(OH)2 (9.00 kg, 150.35 mol, 3.0 equiv), Cu(OAc)2 (10.10 kg, 50.17 mol, 1.0 equiv), 2,2'-dipyridine (7.90 kg, 50.17 mol, 1.0 equiv), anhydrous Na2CO3 (16.00 kg, 150.96 mol, 3.0 equiv), and 4Å molecular sieve (43.60 kg, 4 w / w) were added to the reactor. The reaction mixture was bubbled with 5% O2 / 95% N2 (10–14 L / min) while maintaining the temperature at 20 ± 10 °C. The reactants were heated to 30–35 °C and stirred at that temperature with 5% O2 / 95% N2 for 15 h. Samples were taken for HPLC analysis.

[0224] The reaction mixture was cooled to 20 °C. The reaction mixture was filtered and the filter cake was washed with MTBE (10⁹ L, 10 V). The filtrate was transferred to a reactor and MTBE (10⁹ L, 10 V) and H₂O (10⁹ L, 10 V) were added. The phases were separated and the aqueous phase was extracted with MTBE (2 x 10⁹ L, 2 x 10 V). The combined organic phases were washed with NaCl aqueous solution (5 x 10⁹ L, 5 x 10 V) and H₂O (163.50 L, 15 V), respectively. The organic phases were concentrated to 6–7 V under reduced pressure (≤ 45 °C). The concentrated solution of the product was used in the next step (92.49% a / a, 15.67% HPLC analysis, 9.9 kg product, 78.8% yield, 99.6% ee, 100% de).

[0225] Part 4b -Alternative Synthesis of (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylate: A reactor was charged with (2R,3R)-3-cyclopropylaziridine-2-carboxylate (10 g, 0.046 mol, 1.0 equiv), acetonitrile (118.5 g, 12 x), trimethylcycloboroxane (8.09 g, 50% THF solution, 0.7 equiv, 0.032 mol), Cu(OAc)₂ (8.36 kg, 0.046 mol, 1.0 equiv), 2,2'-dipyridine (7.2 g, 0.046 mol, 1.0 equiv), anhydrous K₂CO₃ (12.72 g, 0.092 mol, 2.0 equiv), and di-tert-butylperoxide (6.73 g, 0.046 mol, 1.0 equiv). The reactants were heated to 35°C and stirred at that temperature for 23 h. Samples were taken for HPLC analysis. The reactants were cooled to 20°C. The reaction mixture was filtered and the filter cake was washed with MTBE (37 g, 3.7 x). The filtrate was transferred to a reactor and cooled to 0°C, after which 30% sodium thiosulfate (145.43 g, 14.5 x) was added dropwise. The mixture was concentrated to 10 V under vacuum at 35°C. The residual mixture was extracted with MTBE (111 g, 11.1 x). The organic phase was concentrated to 2 V under vacuum. The concentrated solution of the product was used for the next step (94.0% purity, 94.0% yield).

[0226] LCMS (ESI+): C14H17NO2[M+H]+ Calculated value: 232.1; Experimental value: 232.1. 1H NMR (400 MHz, DMSO-d 6) δ 7.41-7.30 (m, 5H), 5.22-5.07 (m, 2H), 2.26 (s, 3H), 2.20 (d, J= 6.6 Hz, 1H), 1.39 (t,J= 6.9 Hz, 1H) , 0.82‑0.69 (m, 1H) , 0.49‑0.28 (m, 3H) , 0.22‑0.11 (m, 1H).

[0228] Partial 5-synthesis of (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylic acid ½ Ba salt: (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylic acid benzyl ester (1.50 kg, 6.48 mol, 1.00 equiv), H2O (1.50 L, 1 V), and THF (15.00 L, 10 equiv) were charged into a reactor at 25-30°C.V). Cool the reactants to 0-5°C. Add Ba(OH)₂·8H₂O (1.02 kg, 3.24 mol, 0.5 equiv) to the reaction mixture and heat the reactants to 25-30°C. Stir the reactants at 25-30°C for 16 h. Take samples for HPLC analysis.

[0229] Concentrate the reactants under reduced pressure (25-30°C, 50-100 mbar) and exchange the solvent with MeCN (2 x 15.00 L, 2 x 10 V). Add MeCN (15.00 L, 10 V) and stir the mixture at 25-30°C for 16 h. Concentrate the reaction mixture under reduced pressure (25-30°C, 50-100 mbar) and exchange the solvent with MTBE (3 x 15.00 L, 3 x 10 V). MTBE (15.00 L, 10 V) was added, and the mixture was stirred at 25–30 °C for 16 h. The slurry was filtered, and the filter cake was washed with MTBE (2 x 3.00 L, 2 x 2 V). The wet filter cake was dried under reduced pressure (25–30 °C, 50–100 mbar) to give crude (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylic acid ½ Ba salt (1.38 kg, 58.9% w / w analysis). Table 2 summarizes the HPLC method used for this synthesis.

[0230] LCMS (ESI+): C7H11NO2[M+H]+ Calculated value: 142.1; Experimental value: 142.1.

[0231] 1H NMR (400 MHz, methanol-d4) δ 2.38 (s, 3H), 2.11 (d, J = 7.3 Hz, 1H), 1.23–1.12 (m, 1H), 1.00–0.86 (m, 1H), 0.67–0.53 (m, 1H), 0.52–0.40 (m, 2H), 0.37–0.24 (m, 1H).

[0232] Table 2: HPLC method and IPC method used in Example 2:

[0233] Example 3 - Alternative synthetic route for (2R,3R)-3-cyclopropylaziridine-2-carboxylate Item 72 / 123 pages 84 CN 122270438 Part A 1 - Synthesis of (R,E)-N-(cyclopropylmethylene)-2-methylpropane-2-sulfinamide (R)-tert-butyl-sulfinamide (2.00 kg, 16.50 mol, 1.0 equiv), cyclopropaneformaldehyde (1.20 kg, 17.12 mol, 1.05 equiv), 4Å MS (1.60 kg, 0.8 w / w), DCM (10.00 L, 5 V) and pyrrolidine (5.87 g, 0.08 g) were charged into the reactor.mmol). The reaction mixture was stirred at rt for 16 h, at which point HPLC analysis showed that the reaction was complete. After completion, the mixture was filtered through a diatomaceous earth mat and washed with DCM (3.00 L, 1.5 V). The filtrate was concentrated to give crude product (2.89 kg, 99.6% a / a, 94.7% analytical, 95.8% yield).

[0234] LCMS (ESI+): C8H15NOS [M+H]+ Calculated value: 174.1; Experimental value: 174.1.

[0235] H NMR (400 MHz, CDC13) δ 7.42 (d, J= 7.9 Hz, 1H), 1.94 (qt, J= 8.0, 4.6 Hz, 1H), 1.13 (s, 9H), 1.08-1.01 (m, 2H), 0.95‑0.86 (m, 2H).

[0236] Part 2 - Synthesis of (2R,3R)-1-((R)-tert-butylsulfinyl)-3-cyclopropylaziridine-2-carboxylate: Crude product from Part 1 (1.10 kg, 6.35 mol, 1.0 equiv), benzyl bromoacetate (1.89 kg, 8.25 mol, 1.3 equiv), and THF (11.00 L, 10 V) were added to the reactor. The reaction mixture was cooled to -70°C to -75°C. LiHMDS (1.0 M in THF, 6.35 mol, 1.0 equiv) was added dropwise over 2 h at -70°C to -75°C, and the reaction mixture was stirred for 1 h. The reaction mixture was gradually heated to rt and stirred overnight, at which point HPLC analysis showed that the reaction was complete (IPC: starting material ≤ 1% a / a, 210 nm). After completion, the reaction mixture was poured into ice water (6.05 L, 5.5 V). MTBE (11.00 L, 10 V) was added, and the mixture was stirred at 20–25 °C for 10 min. The mixture was allowed to stand for 30 min, and the organic phase was separated. The aqueous phase was extracted with MTBE (2 x 11.00 L, 2 x 10 V). The combined organic phases were washed with NaCl aqueous solution (10 wt%, 11.00 L, 10 V). The organic phase was concentrated under reduced pressure (25–35 °C, 50–100 mbar). The residue was dissolved in MTBE (11.00 L, 10 V) and loaded with DABCO (0.21 kg, 1.90 mol, 0.3 equiv). The mixture was stirred at 20–25 °C for 2 h, and a sample was taken for HPLC analysis (IPC: benzyl bromoacetate = 0% a / a). The mixture was washed with H₂O (11.00 L, 10 V). NaOH aqueous solution (0.1 M, 11.00 L, 10 V) was added to the organic phase.V) and the mixture was stirred at 20–25 °C for 2 h. The reaction mixture was sampled for HPLC analysis. The organic phase was separated and the aqueous phase was extracted with MTBE (10 V). The combined organic phases were washed with NaCl aqueous solution (10 wt%, 11.00 L, 10 V) and H2O (11.00 L, 10 V), respectively. The organic solvent was concentrated under reduced pressure and MTBE (1.10 L, 1 V) was added to the residue. The mixture was cooled to 10 °C and seeded with product crystals, followed by the addition of n-heptane (8.80 L, 8 V) over 2 h. The mixture was cooled to -5 °C and stirred at -5 °C for 16 h. The slurry was filtered and the filter cake was dried to give the product (1.25 kg, 96.4% LCAP, 94.1% analyte, 57.7% yield).

[0237] LCMS (ESI+): C17H23NO3S [M+H]+ Calculated value: 322.2; Experimental value: 322.2.

[0238] 1H NMR (400 MHz, CDCl3) δ 7.43–7.33 (m, 5H), 5.24 (q, J = 12.3 Hz, 2H), 3.41 (d, J = 7.1 Hz, 1H), 2.07 (t, J = 6.9 Hz, 1H), 1.24 (s, 9H), 0.55–0.47 (m, 3H), 0.35–0.26 (m, 1H).

[0239] Part 3 Synthesis of (2R,3R)-3-cyclopropylaziridine-2-carboxylate: (2R,3R)-1-((R)-tert-butylsulfinyl)-3-cyclopropylaziridine-2-carboxylate (0.10 kg, 0.31 mol, 1.0) and MTBE (1.50 L, 15 V) were added to the reactor. The reaction mixture was cooled to -10°C to -5°C. Dioxane containing HCl (4.0 M, 1.56 mol, 5.0 equiv) was added dropwise over 35 min at -10°C to -5°C. The reaction mixture was stirred at -10°C to -5°C for 30 min and samples were taken for HPLC analysis. The reaction mixture was filtered at -10°C to -5°C and the filter cake was washed with pre-cooled MTBE (-20°C, 0.50 L, 5 V) to give a crude product in the form of an HCl salt.

[0240] MTBE (1.00 L, 10 V) and triethylamine (62.90 g, 0.62 mol, 2.0 equiv) were added to the reactor. The solution was cooled to -10°C to -5°C. The solution was then subjected to 10 [units of temperature] at -10°C to -5°C.The crude product HCl salt was added to the reactants within min. The reactants were stirred at -10°C to -5°C for 3 h and then heated to 0-5°C. H₂O (0.5 L, 5 V) was added to the reactants and stirred for 10 min. The organic phase was separated, and the aqueous phase was extracted with MTBE (0.50 L x 2, 5 V x 2). The combined organic phases were concentrated under reduced pressure at 30°C to give the crude product. The crude product was dissolved in MTBE (0.10 L, 1 V) and the solution was cooled to 15°C. Heptane (0.70 L, 7 V) was added dropwise at 15°C for 2 h. The mixture was cooled to -5°C and stirred overnight at -5°C. The slurry was filtered and dried under vacuum to give (2R,3R)-3-cyclopropylaziridine-2-carboxylate (53.30 g, 99.8% LCAP, 78.8% yield) as a grayish-white solid.

[0241] LCMS (ESI+): C13H15NO2[M+H]+ Calculated value: 218.1; Experimental value: 218.2.

[0242] 1H NMR (400 MHz, CDCl3) δ 7.43-7.31 (m, 5H), 5.32-5.20 (m, 2H), 2.73 (d, J= 6.0 Hz, 1H), 1.68 (dd, J= 8.2, 6.0 Hz, 1H), 0.83-0.70 (m, 1H), 0.65-0.56 (m, 1H), 0.54-0.40 (m, 2H), 0.36-0.25 (m, 1H).

[0243] Example 4 - Synthesis procedure for (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate (compound of formula IIIc-1) 74 / 123 pages 86 CN 122270438 Part A - Synthesis of compound IIIc-1: Dibenzyl oxalate (1.00 kg, 3.70 mol, 1.0 equiv) and THF (30.00 L, 30.0 V) were charged into a reactor at 20-25°C as solution A. R5-MgBr (0.22 M in THF, 1.3 equiv) was charged into another reactor at 20-25°C under N2 flow as solution B. Solution A and solution B were mixed in a plug flow reactor (flow rates: A: 250 mL / min, B: 182.2 mL / min, temperature: -75 to -70 °C, residence time in the reactor: 15 s). The reaction was monitored by HPLC, and HPLC analysis indicated that the reaction was complete. The reaction was quenched with a saturated NH4Cl (10.00 L, 10 V) aqueous solution.The mixture was stirred at 20–25 °C for 30 min. The phases were separated, and the organic phase was washed with an aqueous citric acid solution (0.5 wt%, 10.00 L, 10 V). The organic phase was concentrated to dryness under vacuum, and isopropanol (2.00 L, 2 V) was added to the residue to obtain an IPA solution of the product. An aqueous NaHSO3 solution (40 wt%, 1.50 L, 1.5 V) was added to the reactor, followed by the IPA solution of the product. The mixture was stirred at 20–25 °C for 2 h. IPA (13.5 V) was added to the mixture over 2 h. The mixture was stirred at 20–25 °C for another 10 h. The slurry was filtered, and the filter cake was washed with IPA (3 V). The filter cake was dissolved in H2O (7 V), and an aqueous Na2CO3 solution (20 wt%, 2–2.5 V) was added dropwise to adjust the pH to ≥ 9, while stirring at -5 °C. MTBE (5 V) was added to the solution and the mixture was stirred at rt for 30 min. The organic phase was separated and washed with aqueous citric acid (0.3 V) and brine (5 V), respectively. The organic phase was concentrated to dryness under vacuum to give a colorless oily compound of formula IIIc-1 (99.4% LCAP, 96.8% analyte, 58% yield).

[0244] Part 2-synthetic synthesis of formula IIId-1 compound: H2O (2.50 L, 10 V), Na2HPO4 (84.80 g, 33.9% w / w), and NaH2PO4 (2.00 g, 0.82% w / w) were charged into a reactor at 20-25 °C. The reactants were heated to 30-35 °C. Glucose (0.58 kg, 3.23 mol, 3.0 equiv) and NADP (6.25 g, 2.5% w / w) were added to the reaction mixture. EW-KRED-R122 (5.00 g, 2% w / w) and GDH (12.50 g, 5% w / w) were added to the reaction mixture. The reaction mixture was stirred at 30–35 °C for at least 10 min. After 10 min, n-heptane (0.25 L, 1 V) containing compound IIIc-1 (1.08 mol, 1.0 equiv) was added dropwise to the reaction mixture, and the reaction mixture was stirred at 30–35 °C for 20 h. During this period, the pH was checked after 8 h (the pH was controlled at 7.3–8.2 with 1M NaOH aqueous solution). The reaction was monitored by HPLC, at which point HPLC analysis showed that the reaction was complete. (Reaction manual 75 / 123 pages 87 CN 122270438 A) After completion, MTBE (5.00 L, 20 V) was added, and the mixture was stirred for 10 min. The mixture was filtered and treated with MTBE (0.50 L, 2...V) Wash the filter cake. Let the filtrate stand for 10 min and separate the organic phase. Wash the aqueous phase with MTBE (5.00 L, 20 V). Wash the combined organic phase with NaCl aqueous solution (20 wt%, 5.00 L, 20 V). Concentrate the organic phase to dryness under vacuum to give a colorless oily compound of formula IIId-1 (97.8% LCAP, 97.2% analytical, 100% ee, 97.5% yield).

[0245] Partial 3-synthetic IIIe-1 compound: Compound of formula IIId-1 (83.18 mol, 1.0 equiv) and DCM (234.00 L, 12 V) were charged into the reactor at 15-25 °C, followed by the addition of DIPEA (21.50 kg, 166.36 mol, 2.0 equiv). The reaction mixture was cooled to -10-0 °C under N2 flow. Tf₂O (31.85 kg, 116.45 mol, 1.4 equiv) was added dropwise to the reaction mixture over 1 h. The reaction mixture was stirred at -10–0 °C for 1 h and a sample was taken for HPLC analysis. H₂O (19.50 L, 1 V) was added to the reactants at 0–5 °C, followed by compound I (1 / 2 tartrate, 91.92 mol, 1.1 equiv) and K₃PO₄ (45.90 kg, 216.23 mol, 2.6 equiv). The reactants were heated to 20–25 °C over 1 h and stirred for 6 h. After this, an aqueous HCl solution (0.5 M, 293.00 L, 15 V) was added to the reactants at 10 ± 5 °C. The mixture was stirred for 30 min and the organic phase was separated. H₂O (195.00 L, 10 V) was added to the organic phase at 10 ± 5 °C. The mixture was stirred for 30 min and the organic phase was separated. The organic phase was concentrated to dryness to give a yellow oily product (93.2% LCAP, 91.6% yield). Table 3 summarizes the HPLC method used for this synthesis.

[0246] Table 3: HPLC method IPC method:

[0247] Part 4 - Synthesis of Formula III-1 compound The reactor was charged with Formula IIIe-1 compound (68.00 mol, 1.0 equiv) and EtOAc (150.50 L, 5 V). The solution was cooled to 0-5°C. An EtOAc solution containing HCl (4 M, 150.50 L, 5 V) was added dropwise to the reactants. The reactants were heated to 20-30°C and stirred at 20-30°C for 6 h. The reaction was monitored by HPLC, and the HPLC analysis showed that the product was 76 / 123 pages, 88 CN 122270438 A.The reaction was completed. After completion, the reactants were cooled to 0–10 °C, and an aqueous HCl solution (0.5 M, 301.00 L, 10 V) was added to the reactants. The mixture was stirred at 20–30 °C for 20 min, allowed to stand, and the phases were separated. The organic phase was extracted with an aqueous HCl solution (0.5 M, 301.00 L, 10 V) at 0–20 °C. The combined aqueous phases were washed with MTBE (2 x 301.00 L, 2 x 10 V). The pH of the aqueous phase was adjusted to 11–12 with an aqueous NaOH solution (20 wt%) at 5–10 °C. The aqueous phase was extracted with MTBE (2 x 301.00 L, 2 x 10 V). The combined organic phases were washed with an aqueous NaCl solution (26 wt%, 301.00 L, 10 V). The organic phase was concentrated to dryness under vacuum to give a crude, colorless oily compound of formula III-1 (analytical yield 90.1%).

[0248] Crude compound of formula III-1 (61.29 mol, 1.0 equiv) and IPA (210.00 L, 10 V) were added to a reactor at 20 ± 10 °C. In another reactor, a solution of D-(+)-2,3-dibenzoyl tartaric acid (21.96 kg, 61.29 mol, 1.0 equiv) in IPA solution (210.00 L, 10 V) was prepared. 1 / 5 of the IPA solution of D-(+)-2,3-dibenzoyl tartaric acid (42.00 L, 2 V) was added dropwise to the solution over 1.5 h at 40–45 °C. Seed crystals of compound III-1 (0.01 w / w) were added to the reactants at 40–45 °C. The mixture was stirred at 40–45 °C for 1.5 h. After 6 h, the remaining (4 / 5) of the IPA solution containing D-(+)-2,3-dibenzoyl tartaric acid (168.00 L, 8 V) was added dropwise to the reactants. The mixture was stirred at 40–45 °C for 3 h and then cooled to 0–5 °C. The mixture was stirred at 0–5 °C for another 6 h. The slurry was filtered, and the filter cake was washed with IPA (42.00 L, 2 V) and MTBE (42.00 L, 2 V). The filter cake was dissolved in H₂O (210.00 L, 10 V), and MTBE (420.00 L, 20 V) was added to the solution. The pH of the aqueous phase was adjusted to 11–12 with NaOH aqueous solution (10 wt%). The mixture was stirred for 45 min, then allowed to stand, and the phases were separated. The organic phase was washed with NaCl aqueous solution (26 wt%, 210.00 L, 10 V). The organic phase was concentrated to dryness under vacuum to give a colorless oily product (99.96% LCAP, 99.01% ee, 85.8% yield). Table 4 summarizes the HPLC method used for this synthesis.

[0249] Table 4: HPLC Method IPC Method:

[0250] Example 5 - Alternative Synthesis of Formula IIId-1 Compound Part 1 - Synthesis of Formula IIIg Compound Specification 77 / 123 pages 89 CN 122270438 A H2SO4 (56.76 kg, 578.78 mol, 2.5 equiv) and H2O (660.00 L, 20 V) were charged into the reactor at 20-25°C, followed by the addition of Formula IIIf compound (231.51 mol, 1.0 equiv). The reaction mixture was stirred at 20-25°C for 30 min to form a homogeneous solution. The solution was filtered through a microporous filter (0.22 µm) into tank A as solution A. NaNO2 (102.45 kg, 1484.88 mol, 6.4 equiv) and H2O (800.00 L, 24.1 V) were charged into another reactor at 20–25 °C. The reaction mixture was stirred at 20–25 °C for 30 min and transferred to tank B as solution B. Solutions A and B were mixed using a continuous microreactor. The flow rates were 37.7 L / h for A and 27.7 L / h for B, the temperature was 50 °C, and the retention time in the microreactor was 2 min. The solution was purged with N2 gas for 12 h to push most of the NO2 gas into the exhaust gas absorption reactor. NaCl (198.9 kg, 6 w / w) was charged into the reactor and stirred to dissolve the solids. The solution was extracted with MTBE (2 x 994.50 L, 2 x 30 V). The organic phase was concentrated to 2–3 V under reduced pressure (0.08 MPa, 20–30 °C). The MTBE residue was exchanged with n-heptane at reduced pressure (0.08 MPa, 20–30 °C) to 2–3 V until the MTBE residue was less than 3 wt% and KF was less than 0.5 wt%. EtOAc (19.90 L, 0.6 V) was added to the n-heptane residue, and the mixture was heated to 45 ± 2 °C. The reaction mixture was stirred at 45 ± 2 °C for at least 8 h and then gradually cooled to 30 °C at a rate of 1.5 °C / h. The slurry was filtered, and the filter cake was washed with EtOAc / n-heptane (1:5 v / v, 33.00 L, 1 V). The wet filter cake was dried under vacuum (0.09 MPa, 45 ± 5 °C) to give compound IIIg as a grayish-white solid (91.4% LCAP, 44.9% yield). Table 5 summarizes the HPLC method used for this synthesis.

[0251] Table 5: HPLC method IPC method:

[0252] Part of the 2-synthetic IIId-1 compound was charged into the reactor at 20-25°C under N2 atmosphere with dimethylacetamide (128.00 L, 5 V) and DIPEA (34.44 L).The reaction mixture was prepared by adding BnBr (36.45 kg, 213.11 mol, 1.2 equiv) and compound III (177.57 mol, 1.0 equiv). The reaction mixture was cooled to 0–10 °C and BnBr (36.45 kg, 213.11 mol, 1.2 equiv) was added dropwise. The reaction mixture was heated to 15–25 °C and stirred for 6 h. The reaction was monitored by GC, at which point GC analysis indicated completion. DMAP (13.01 kg, 106.49 mol, 0.6 equiv) was added, and the mixture was stirred at 20–25 °C for 18 h. Samples were taken for GC analysis to ensure that BnBr was not detected. H2O (256.00 L, 10 V) was added to the solution, and the resulting solution was extracted with MTBE (2 x 128.00 L, 2 x 5 V). The combined organic phases were washed with aqueous citric acid (2 x 128.00 L, 2 x 5 V) and H₂O (128.00 L, 5 V), respectively. The organic phases were concentrated to 2-3 V under reduced pressure (0.08 MPa, 20–35 °C). The MTBE residue was exchanged to 2-3 V with DCM (256.00 L, 10 V) under reduced pressure (0.08 MPa, 20–35 °C) until GC analysis showed that the MTBE residue was less than 3% LCAP. A solution of compound IIId-1 in DCM was obtained (95.6% LCAP, 105% yield). Table 6 summarizes the HPLC method used for this synthesis.

[0253] Table 6: HPLC method IPC method:

[0254] Example 6 - Synthesis procedure of Formula IV compound Part 1 - Synthesis of Formula IV-1 compound At 15±10°C, the barium salt of compound A (8.5 kg, 40.88 mol, 1.0 equiv) was dissolved in DMF (56.00 L, 4 V) in a reactor. NMI (13.4 kg, 163.22 mol, 4.0 equiv) and Formula III-1 compound (40.88 mol, 1.0 equiv) were added to the solution, followed by dropwise addition of TCFH (13.80 kg, 49.18 mol, 1.2 equiv) to the solution in DMF (4.00 L, 1 V) over 1 h at 15±10°C. The reaction mixture was stirred at 15±10℃ for at least 30 min. The reaction was monitored by HPLC, at which point HPLC analysis showed that the reaction was complete.

[0255] The reaction mixture was cooled to 5±5℃ and treated with H2O (28.00 L, 2(V) Quenching. The mixture was heated to 20 ± 5 °C and stirred for 10 min. MTBE (280.00 L, 20 V), H2O (182.00 L, 13 V) and Na2CO3 (1.12 kg, 0.08 w / w) were added to the mixture and stirred for 20 min. The organic phase was separated, and the aqueous phase was extracted with MTBE (140.00 L, 10 V). The combined organic phases were washed with an aqueous solution of N2CO3 (15 wt%, 2 x 164.60 kg, 2 x 11.76 w / w). The organic phase was concentrated to 4-5 V under reduced pressure (0.08 MPa, 20–40 °C). The process was repeated twice by exchanging MTBE with EtOAc (280.00 L, 20 V) and concentrating to 4-5 V under reduced pressure (0.08 MPa, 20–40 °C). A portion of EtOAc (210.00 L, 15 V) was added to the residue and the mixture was stirred for 40 min. The solution was cooled to -5 ± 5 °C and maleic acid (2.37 kg, 20.42 mol, 0.5 equiv) was added to the solution in portions. Maleate seed crystals of compound IVa-1 were added to the reactants and stirred for 15 min. After 7.5 h, a solution of maleic acid (6.17 kg, 1.3 equiv) in EtOAc (280.00 L, 20 V) was added dropwise to the reactants and stirring was continued at -5 ± 5 °C for 7.5 h. The slurry was filtered and the filter cake was washed with pre-cooled EtOAc (42.00 L, 3 V) at -5 ± 5 °C. The wet filter cake was collected.

[0256] MTBE (140.00 L, 10 V) and the wet filter cake were added to the reactor at 0 ± 5 °C. Add 15 wt% Na₂CO₃ aqueous solution at 0±5 °C until the pH of the aqueous layer is 9–11. Heat the mixture to 25±5 °C and stir for 30 min. Separate the organic phase and extract the aqueous phase with MTBE (140.00 L, 10 V). Wash the combined organic phases with Na₂CO₃ aqueous solution (15% wt%, 140.00 L, 10 V). Concentrate the organic phase to 4–5 V under reduced pressure (0.08 MPa, 20–40 °C). Exchange MTBE with MeOH (280.00 L, 20 V) and concentrate to 4–5 V under reduced pressure (0.08 MPa, 20–40 °C), repeating this process twice. Add a portion of MeOH (112.00 L, 8 V) to the residue and stir the mixture at 20±10 °C for 30 min. Take a sample for GC analysis (MTBE residue ≤ 3 wt%). A MeOH solution (97.8%) of compound IVa-1 was obtained.LCAP, 50.7% yield).

[0257] Part 2 - Synthesis of Compound IV: A MeOH solution of compound IVa-1 (13.00 mol, 1.0 equiv) and TEA (4.00 kg, 39.00 mol, 3.0 equiv) were added to the reactor. The mixture was cooled to 0±5°C and PdCl2 (60.50 g, 1% w / w) was added to the reactants. The reaction mixture was stirred at 0±5°C and 0-0.15 MPa H2 pressure for at least 3 h. Samples were taken for HPLC analysis. The reaction mixture was filtered through diatomaceous earth and the filter cake was washed with MeOH (42.35 L, 7 V). The filtrate was concentrated to 8-10 V under reduced pressure (20-30°C, 0.08 MPa). The solvent was exchanged twice under reduced pressure (20–30 °C, 0.08 MPa) with THF (60.50 L, 10 V) to 8–10 V. THF (60.50 L, 10 V) and TEA (2.00 kg, 19.76 mol, 1.5 equiv) were added to the residue. The resulting solution was concentrated to 8–10 V under reduced pressure (20–30 °C, 0.08 MPa), and THF (18.15 L, 3 V) was added to the residue. The solution was stirred at 20 ± 10 °C for at least 30 min to form a slurry. MTBE (60.50 L, 10 V) was added to the slurry at 20 ± 10 °C for 1.2 h. The mixture was concentrated to 10–13 V under reduced pressure (20–30 °C, 0.08 MPa). The solvent was changed by adding MTBE dropwise over 1 h and then concentrating under reduced pressure (20–30 °C, 0.08 MPa), followed by solvent exchange with MTBE (48.40 L, 8 V) to 10–13 V six times. MTBE (48.40 L, 8 V) was added dropwise to the suspension over 1 h at 20 ± 10 °C, and samples were taken for analysis (THF residue ≤ 7% w / w). The slurry was filtered, and the filter cake was washed with (18.15 L, 3 V). The filter cake was dried under vacuum (30 ± 5 °C, 0.08 MPa) for at least 8 h until LOD ≤ 10% w / w. Compound of formula IV was obtained as a grayish-white solid (95.9% LCAP, 90.2% w / w analysis, 76.4% yield).

[0258] A MeOH solution of compound IV (1.0 equiv) and TEA (3.0 equiv) were added to the reactor. The mixture was cooled to 0 ± 5 °C and PdCl2 (1% w / w) was added to the reactants. The reaction mixture was then subjected to a pressure of 0–0.15 MPa at 0 ± 5 °C.Stirring under H2 pressure for at least 3 h. Samples were taken for HPLC analysis. The reaction mixture was filtered through diatomaceous earth and the filter cake was washed with MeOH (7 V). The filtrate was concentrated to 3–4 V under reduced pressure (20–30 °C, 0.08 MPa). MeOH (7 V) was added to the residue and the solution was stirred at rt for 10 min. NaOMe (1.03 equiv, 30 wt% MeOH solution) was added to the solution at rt and the resulting mixture was stirred at rt for at least 3 h. The reaction solution was concentrated to 3–4 V under reduced pressure (20–30 °C, 0.08 MPa) to obtain solution A. MTBE (60 V) was loaded into another reactor and the reactor was cooled to -5–5 °C. Seed crystals (2 wt%) of the Na salt of formula IV were added to the MTBE solution and the suspension was stirred for 30 min. Solution A of formula IV was added to MTBE at -5–5 °C for 5 h and the suspension was stirred for 2 h. The slurry was filtered, and the wet filter cake was washed with MTBE (2 V). The wet filter cake was dried under vacuum at 30–40 °C for 5 h to obtain the Na salt of compound IV. Table 7 summarizes the HPLC method used for this synthesis.

[0259] Table 7: HPLC Method IPC Method:

[0260] Example 7 - Alternative Synthetic Route of Formula IVa-1 Compound Specification 81 / 123 pages 93 CN 122270438 A MTBE (90.50 mL, 5 V), Formula III-1 compound (52.85 mmol, 1.0 equiv), (2R,3R)-3-cyclopropyl-1-methylaziridine-2-carboxylate benzyl ester (12.84 g, 55.49 mmol, 1.05 equiv), and 1,5,7-triazabicyclo[4,4,0]dec-5-ene (TBD) (2.21 g, 15.86 mmol, 0.3 equiv) were charged into the reactor. The reaction mixture was cooled to -5 to 0 °C. The reaction mixture was stirred at -5 to 0 °C for 24 hours, at which time HPLC analysis showed that the reaction was complete. Add an aqueous solution of Na₂CO₃ (5 wt%, 181.00 mL, 10 V) to quench the reaction and stir the mixture at 15–25 °C for at least 10 min. Separate the phases and wash the organic phase with an aqueous solution of Na₂CO₃ (5 wt%, 181.00 mL, 10 V). Concentrate the organic phase to dryness (T ≤ 40 °C). Dissolve the residue in EtOAc (145.00 mL, 8 V) and cool the solution to -10–0 °C. Add maleic acid (3.07 g, 26.42 mmol, 0.5 equiv) to the solution, followed by the addition of maleate seed crystals of compound IVa-1 at -10–0 °C.(wt%). The mixture was stirred for at least 10 min, followed by dropwise addition of maleic acid to a solution of MTBE / EtOAc (v / v = 3:2, 181.00 mL, 10 V) over 3 h. The suspension was stirred at -10–0 °C for 12 h. The slurry was filtered, and the filter cake was washed with pre-cooled MTBE / EtOAc (v / v = 1:2, 54.30 mL, 3 V). The filter cake was transferred to a reactor where MTBE (181.00 mL, 10 V) was cooled to -5–0 °C. An aqueous solution of Na₂CO₃ (15 wt%, 181.00 mL, 10 V) was added to the solution to adjust the pH to 8–10. The mixture was stirred at 15–25 °C for at least 10 min. The phases were separated, and the aqueous phase was extracted with MTBE (90.50 mL, 5 V). The organic phase was washed with an aqueous solution of Na₂CO₃ (5 wt%, 181.00 mL, 10 V). The organic phase was concentrated to dryness to obtain maleate of formula IVa-1 (97.2% LCAP, 92.7% QNMR analysis). Table 8 summarizes the HPLC method used for this synthesis.

[0261] Table 8: HPLC method IPC method:

[0262] Example 8 - Synthesis procedure of compound B - (S)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester hemiester ((2R,3R)-2,3-dihydroxysuccinate) 82 / 123 pages 94 CN 122270438 Part 1 - Synthesis of 3-(cyanomethyl)pyrrolidine-1,3-dicarboxylic acid 1-(tert-butyl) ester 3-methyl ester THF (16.00 L, 8 V) was charged into the reactor. The reactor was cooled to -20 to -15°C. n-BuLi (2.5 M in hexane, 4.54 L, 1.3 equiv) was added dropwise to the reactor while maintaining the internal temperature between -20 and -15 °C. Then, diisopropylethylamine (DIPEA, 1.24 kg, 12.25 mol, 1.4 equiv) was added dropwise at -20 to -15 °C. The reaction mixture was stirred under N2 at -20 to -15 °C for 30 min. The reaction mixture was cooled to -70 to -80 °C. At -70 to -80 °C, THF (2.00 L, 1 V) containing 3-methyl 1-(tert-butyl) pyrrolidine-1,3-dicarboxylate (2.00 kg, 8.72 mol, 1.0 equiv) was added dropwise to the reaction mixture. The reaction mixture was stirred under N2 at -70 to -80 °C for 30 min. At -70 to -80°C, THF (2.00 L, 1 mol, 1 equiv) containing 2-bromoacetonitrile (1.36 kg, 11.34 mol, 1.3 equiv) was added.V) was added dropwise to the reaction mixture. The reaction mixture was heated to 20–30 °C. The reaction was monitored by HPLC. The reaction mixture was quenched dropwise by adding 15% w / w NH4Cl aqueous solution (16.00 L, 8 V) while maintaining the temperature at 20 ± 10 °C. The mixture was extracted with EtOAc (20.00 L, 10 V) and EtOAc (10.00 L, 5 V). The combined organic layers were washed with 15% w / w NaCl aqueous solution (8.00 L x 3, 4 V). The organic layers were then concentrated (40–50 °C, 100 ± 50 mbar) to give a deep red oily 3-(cyanomethyl)pyrrolidine-1,3-dicarboxylic acid 1-(tert-butyl) ester 3-methyl ester (7.70 kg, 99.66% a / a purity, 51.3% w / w analysis, 60% yield). LCMS (ESI): C13H20N2O4[M+H2O]+ Calculated value: 286.2; Experimental value: [M+H2O]: 286.2. 1H NMR (300 MHz, DMSO-d 6, 25℃) δ 3.71 (s, 3H), 3.68–3.64 (d, J = 12 Hz, 1H), 3.38–3.25 (m, 3H), 3.00 (t, J = 2.1 Hz, 2H), 2.31–2.21 (m, 1H), 2.02–1.90 (m, 1H), 1.90 (s, 9H). Table 9 summarizes the HPLC method used for this synthesis.

[0263] Table 9: HPLC method for Part 1 HPLC method: Specification 83 / 123 pages 95 CN 122270438 A

[0264] Part 2 - Synthesis of tert-butyl 6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate MeOH (59.00 L, 10 V), 3-(cyanomethyl)pyrrolidine-1,3-dicarboxylate 1-(tert-butyl) ester 3-methyl ester (2) (5.90 kg, 21.99 mol, 1.0 equiv) and Raney Ni (5.90 kg, 100% w / w) were charged into the reactor. The reaction mixture was purged three times with N2 gas, followed by three times with H2 gas to reach 1 atm. The reaction mixture was stirred at 20-30 °C for 16 hours, at which time HPLC analysis showed that the reaction was complete (IPC: 2 / (2+3) ≤ 1% a / a, 210 nm). The mixture obtained by filtration through diatomaceous earth (5.90 kg, 100% w / w) was washed with MeOH (23.60 L, 4 V) and H₂O (23.60 L, 4 V), respectively. The filtrate was concentrated (40–50 °C, 80–150 mbar) to 4–5 ppm.Volume V. Add DCM (47.20 L, 8 V) to the residue. Separate the organic layer and extract the aqueous layer with DCM (23.60 L, 4 V). Dry the combined organic layers with anhydrous Na2SO4 (11.80 kg) at 20–30 °C for 30 min. Filter the organic phase and concentrate under reduced pressure to 2 V (40–50 °C, 80–150 mbar). Add MTBE (11.80 L, 2 V) to the residue and concentrate the mixture to obtain solids (40–50 °C, 80–150 mbar). Add MTBE (35.40 L, 6 V) to the residue and stir the mixture at 25 ± 5 °C for 1 h. Add n-heptane (11.80 L, 2 V) dropwise to the mixture and stir the mixture at 25 ± 5 °C for 3–5 h. The resulting slurry was filtered and the filter cake was washed with n-heptane / MTBE (v / v, 1:1, 5.90 L, 1 V). The wet filter cake was dried under reduced pressure (40–50 °C, 80–150 mbar) for 16 h. A yellow solid, tert-butyl 6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (2.25 kg, 98.9% a / a purity, 97% w / w analysis, 61% yield) was obtained. LCMS (ESI): C12H20N2O3[M- tBu+H] Calculated value: 185.1; Experimental value: [M-tBu+H]: 185.2. 1H NMR (300 MHz, DMSO-d 6, 25℃) δ 6.38 (s, 1H), 3.66–3.25 (m, 6H), 2.23–2.12 (m, 3H), 1.80–1.77 (m, 1H), 1.45 (s, 9H). Table 10 summarizes the HPLC method used for this synthesis.

[0265] Table 10: HPLC method for Part 2 HPLC method: Specification 84 / 123 pages 96 CN 122270438 A

[0266] Part 3 - Synthesis of tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate Toluene (10.00 L, 5 V), tert-butyl 6-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (2.00 kg, 8.32 mol, 1.0 equiv), NiCl2 (DME) (0.013 kg, 0.058 mol, 0.007 equiv) and PhSiH3 (1.98 kg, 18.30 mol, 2.2 equiv) were charged into a sealed autoclave reactor. The reaction mixture was purged three times with N2 gas. The reactants were stirred at 115–120 °C for 24 h. HPLC analysis at this time showed that the reaction was complete (IPC: 3 / (3+4) ≤ 2%).(a / a, 210 nm). Cool the reaction mixture to room temperature. Add MTBE (20.00 L, 10 V) and aqueous citric acid solution (20% w / w, 12.00 L, 6 V). Stir the mixture at 20–30 °C for 2–3 h. Separate the aqueous layer and extract the organic layer with aqueous citric acid solution (20% w / w, 2.00 L, 1 V). Add MTBE (10.00 L, 5 V) to the combined aqueous phases. Separate the aqueous layer and adjust the pH to 13–14 with aqueous KOH solution (20% w / w, 14.00 L, 7 V). Extract the aqueous phase with MTBE (30.00 L x 4, 15 V). Dry the combined organic phases with anhydrous Na₂SO₄ and filter. The filtrate was concentrated under reduced pressure (40–50 °C, 50–300 mbar) to obtain tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (1.95 kg, 97.7% a / a purity, 80.8% w / w analysis, 86% yield), which was a brown oil. LCMS (ESI+): C12H22N2O2[M+H]+ Calculated value: 227.2; Experimental value: 227.1. 1H NMR (400 MHz, CDCl3) δ 3.52–3.31 (m, 2H), 3.31–3.16 (m, 2H), 3.01 (t, J = 7.1 Hz, 2H), 2.85 (dd, J = 11.0, 6.0 Hz, 1H), 2.77 (dd, J = 11.0, 4.5 Hz, 1H), 1.99 (s, 1H), 1.87–1.64 (m, 4H), 1.46 (s, 9H). Table 11 summarizes the HPLC method used for this synthesis.

[0267] Table 11: HPLC method for part 3 HPLC method: Specification 85 / 123 pages 97 CN 122270438 A

[0268] Part 4 - (S)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester ½ tartaric acid (compound B ½ tartrate) The reactor was charged with the racemic compound B (1.90 kg, 8.40 mol, 1.0 equiv), EtOH (18.24 L, 9.6 V), L(+)-tartaric acid (0.63 kg, 4.20 mol, 0.5 equiv) and H2O (0.66 L, 0.35 V). The reaction mixture was purged three times with N2 gas and then stirred at 80 °C for 3 h. The reaction mixture was then slowly cooled to 25 °C over 12 h. The reaction mixture was stirred at 20–25 °C for 4 h. The slurry was filtered and the filter was washed with EtOH (1.90 L, 1 V).The wet filter cake was loaded into a reactor with EtOH (5.70 L, 3 V) and H₂O (0.20 L, 0.105 V). The mixture was purged three times with N₂ gas and then stirred at 80 °C for 3 h. The reactants were then slowly cooled to 25 °C over 12 h. The reaction mixture was stirred at 20–25 °C for 4 h. The slurry was filtered, and the filter cake was washed with EtOH (1.90 L, 1 V) and MTBE (1.90 L, 1 V), respectively. The filter cake was dried under reduced pressure (40 °C, 50–80 mbar) to give compound B½ tartrate as a grayish-white solid (0.979 kg, 100% a / a purity, 93.3% ee, 39% yield). LCMS (ESI+): C12H22N2O2[M+H]+ Calculated value: 227.2; Experimental value: 227.1. 1H NMR (400 MHz, D2O) δ 4.28 (s, 1H), 3.44 (dt, J = 12.1, 7.4 Hz, 3H), 3.40–3.30 (m, 3H), 3.25 (s, 2H), 2.02 (dt, J = 11.0, 7.0 Hz, 2H), 1.96 (td, J = 7.0, 3.0 Hz, 2H), 1.42 (s, 9H). Tables 12 and 13 summarize the HPLC methods used for this synthesis.

[0269] Table 12: Non-chiral HPLC method for part 4, 86 / 123 pages, 98 CN 122270438 A

[0270] Table 13: Chiral HPLC method for part 4

[0271] Example 9 - Substitutional synthesis of compound B - (S)-2,7-diazaspiro[4.4]nonane-2-tert-butyl carboxylate hemi((2R,3R)-2,3-dihydroxysuccinate) - Synthesis of methyl 5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate, 87 / 123 pages, 99 CN 122270438 A MeOH (10 L, 4 V), (S)-(-)-1-phenylethylamine (2.50 kg, 20.63 mol, 1.0 equiv), and dimethyl itaconic acid (3.26) were charged into the reactor. kg, 20.61 mol, 1.0 equiv). The reaction mixture was stirred at 60–70 °C for 22 h. After completion, the reaction mixture was concentrated under vacuum to remove MeOH. Toluene (12.5 L, 5 V) was added to the residue and the mixture was stirred at 100–110 °C for 20 h. The mixture was cooled to 20–25 °C and an aqueous solution of citric acid (20% w / w, 1.5 kg, 20.61 mol, 1.0 equiv) was added.L). The mixture was stirred at 20-25°C for 3-5 h. The organic layer was separated and concentrated under vacuum to give methyl 5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate (5.1 kg) as an orange oil.

[0272] Part 2-Synthesis of (R)-3-(cyanomethyl)-5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate THF (16 L, 10 V) and methyl 5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate (1.70 kg, 6.87 mol, 1.0 equiv) were added to the reactor. The mixture was cooled to -75 to -85°C. LiHMDS (1.0 M, 1.15 kg, 1.0 equiv) was added and the reaction mixture was stirred at -75 to -85°C for 2 h. Add BrCH2CN (0.82 kg, 6.84 mol, 1.0 equiv) and stir the reaction mixture at -75 to -85 °C for 2 h. After completion, add HOAc (0.82 kg, 13.65 mol, 2.0 equiv) at -15 to 20 °C. Concentrate the mixture and add EtOAc (5.1 L, 3 V). Add an aqueous solution of NaCl (25% w / w, 2.5 L, 1.5 V) to the mixture and stir for 0.5 h. Separate the organic phase and concentrate. Add MTBE (3.5 L, 2 V) to the residue and stir the mixture for 2–4 h. Filter the resulting slurry and collect the wet filter cake. Dry the wet filter cake to give methyl (R)-3-(cyanomethyl)-5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate (0.63 kg) as an orange powder.

[0273] Part 3-Synthesis of (R)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-1,8-dione: MeOH (4.5 L, 5 V) and Raney Ni (0.225 kg, 25% w / w) were charged into the reactor at 15-25 °C. Methyl (R)-3-(cyanomethyl)-5-oxo-1-((S)-1-phenylethyl)pyrrolidine-3-carboxylate (0.90 kg, 3.14 mol, 1.0 equiv) and TEA (0.32 kg, 3.16 mol, 1.0 equiv) were added to the mixture. The mixture was degassed with N2 gas and then H2 (1.0 MPa) was charged into the reactor. The reactants were stirred at 50–60 °C for 24 h under a H2 atmosphere. Afterward, the mixture was filtered and the filtrate was concentrated. 2-MeTHF (0.8 L, 1 V) and n-heptane (0.4 L) were added to the concentrated residue.L, 0.5 V). The mixture was stirred at 60-70°C for 2-4 h. The mixture was cooled and the slurry was filtered. The wet filter cake was dried to obtain (R)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-1,8-dione (0.62 kg) in the form of a yellow solid.

[0274] Partial 4-synthesis of (S)-2-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane 2-MeTHF (6.0 L, 5 V) and LiAlH4 (2.5 M in THF, 3.72 L, 9.29 mol, 2.0 equiv) were charged into the reactor. (R)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-1,8-dione (1.2 kg, 4.64 mol, 1.0 equiv) was added dropwise to the reactor at 50–60 °C. The reaction mixture was stirred at 50–60 °C for 12 h. The reaction mixture was cooled to 15–25 °C and diluted with 2-MeTHF (3.6 L, 3 V). H2O (0.35 L, 0.3 V), NaOH aqueous solution (0.35 L, 0.3 V), and H2O (1.06 L, 0.9 V) were added sequentially to the mixture at 15–25 °C. The mixture was stirred for another 1 h. The resulting slurry was filtered and the filtrate was collected as a 2-MeTHF solution containing (S)-2-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane, which was used directly in the next step (0.86 kg for analysis).

[0275] Partial 5-synthetic (S)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-tert-butyl carboxylate was added to the reactor along with a 2-MeTHF solution of (S)-2-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane (0.86 kg for analysis, 3.73 mol, 1.0 equiv) and Boc2O (0.85 kg, 3.9 mol, 1.05 equiv). The resulting mixture was stirred at 15–25 °C for 1 h and then at 45–55 °C for 12 h. The organic solution was washed with an aqueous NaCl solution (25% w / w, 2.4 L, 2.8 V). The organic layer was concentrated to give an orange oily (S)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (1.24 kg).

[0276] Part 6-Synthetic compound B ½ L-tartrate-(S)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester ½ L-tartrate Specification 89 / 123 pages 101 CN 122270438 A IPA (4.2 L, 3.4 V) was added to the reactor.(S)-7-((S)-1-phenylethyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (1.23 kg, 3.72 mol, 1.0 equiv) and L-tartaric acid (0.28 kg, 1.86 mol, 0.5 equiv) were added to the reactor at 15–25 °C with H2O (2.1 L). Pd / C (0.06 kg, 5% w / w) was added to the reaction mixture under N2. The mixture was degassed with H2 (0.5 MPa) and stirred at 15–25 °C for 12 h. The mixture was filtered and the filtrate was collected. The filtrate was concentrated to 1–2 V under vacuum at 40–50 °C. IPA (3.6 L, 3 V) was added to the concentrated residue. The resulting mixture was stirred at 40–50 °C for 2 h. The mixture was cooled to 15 to 25°C and stirred for 16 h. The slurry was filtered and the wet filter cake was collected. The wet filter cake was dried to give compound B ½ tartrate (0.90 kg) as a white solid.

[0277] Example 10 - Synthesis procedure of compound D - (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate Part 1 - Synthesis of 2-cyclopentyl-2-oxobenzyl acetate Dibenzyl oxalate (1.00 kg, 3.70 mol, 1.0 equiv) and THF (30.00 L, 30.0 V) were charged into the reactor at 20-25°C as solution A. Cyclopentylmagnesium bromide (0.22 M in THF, 4.81 mol, 1.3 equiv) was added to another reactor as solution B under N2 flow at 20–25 °C. Solutions A and B were mixed in a plug flow reactor (flow rate: A: 250 mL / min, B: 182.2 mL / min, temperature: -75 to -70 °C, retention time in the reactor: 15 s). The reaction was monitored by HPLC, and HPLC analysis showed that the reaction was complete. The reaction was quenched with a saturated NH4Cl (10.00 L, 10 V) aqueous solution. The mixture was stirred at 20–25 °C for 30 min. The phases were separated, and the organic phase was washed with an aqueous citric acid solution (0.5 wt%, 10.00 L, 10 V). The organic phase was concentrated to dryness under vacuum, and isopropanol (2.00 L, 2 V) was added to the residue to obtain a 2-cyclopentyl-2-oxoacetic acid benzyl ester IPA solution. A NaHSO3 aqueous solution (40 wt%, 1.50 L, 1.5 V) was added to the reactor, followed by a 2-cyclopentyl-2-oxoacetic acid benzyl ester (IPA) solution. The mixture was stirred for 2 h at 20–25 °C. During this 2 h, IPA (13.50 L, 13.5 V) was added.V) was added to the mixture. The mixture was stirred at 20–25 °C for 10 h. The slurry was filtered, and the filter cake was washed with IPA (3.00 L, 3 V). The filter cake was dissolved in H2O (7.00 L, 7 V), and Na2CO3 aqueous solution (20 wt%, 2.00–2.50 L, 2–2.5 V) was added dropwise to the solution to adjust the pH ≥ 9, while stirring at -5 °C. MTBE (5.00 L, 5 V) was added to the solution, and the mixture was stirred at rt for 30 min. The organic phase was separated and washed with citric acid aqueous solution (0.5 wt%, 3.00 L, 3 V) and brine (5.00 L, 5 V), respectively. The organic phase was concentrated to dryness under vacuum to give a colorless oily 2-cyclopentyl-2-oxoacetic acid benzyl ester (543 g, 99.4% LCAP, 96.8% analytical, 58% yield).

[0278] LCMS (ESI+): C14H16O3[M+NH4]+ Calculated value: 250.1; Experimental value: 250.0.

[0279] 1H NMR (400 MHz, DMSO-d6) δ 7.59-7.26 (m, 5H), 5.28 (s, 2H), 3.47 (tt, J= 8.9, 6.8 Hz, 1H), 1.97-1.77 (m, 2H), 1.77-1.63 (m, 2H), 1.63-1.43 (m, 4H).

[0280] Partial 2-Synthesis of (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester: H2O (2.50 L, 10 V), Na2HPO4 (84.80 g, 33.9% w / w), and NaH2PO4 (2.00 g, 0.82% w / w) were charged into a reactor at 20-25°C. The reactants were heated to 30-35°C. Glucose (0.58 kg, 3.23 mol, 3.0 equiv) and NADP (6.25 g, 2.5% w / w) were added to the reaction mixture. EW-KRED-R122 (5.00 g, 2% w / w) and GDH (12.50 g, 5% w / w) were added to the reaction mixture. The reaction mixture was stirred at 30-35°C for at least 10 min. After 10 min, n-heptane (0.25 L, 1 V) containing 2-cyclopentyl-2-oxoacetic acid benzyl ester (0.25 kg, 1.08 mol, 1.0 equiv) was added dropwise to the reaction mixture, and the reaction mixture was stirred at 30–35 °C for 20 h. During this period, the pH was checked after 8 h (using 1 M...The pH was controlled at 7.3–8.2 using NaOH aqueous solution. The reaction was monitored by HPLC, and the HPLC analysis indicated that the reaction was complete. After the reaction was complete, MTBE (5.00 L, 20 V) was added, and the mixture was stirred for 10 min. The mixture was filtered, and the filter cake was washed with MTBE (0.50 L, 2 V). The filtrate was allowed to stand for 10 min, and the organic phase was separated. The aqueous phase was washed with MTBE (5.00 L, 20 V). The combined organic phases were washed with NaCl aqueous solution (20 wt%, 5.00 L, 20 V). The organic phase was concentrated to dryness under vacuum to give (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester (245 g, 97.8% LCAP, 97.2% analytical, 100% ee, 97.5% yield) as a colorless oil.

[0281] LCMS (ESI+): C14H18O3[M+Na]+ Calculated value: 257.1; Experimental value: 257.2.

[0282] 1H NMR (400 MHz, DMSO-d6) δ 7.47-7.27 (m, 5H), 5.38 (d, J= 6.1 Hz, 1H), 5.13 (d, J= 1.2 Hz, 2H), 3.93 (t, J= 6.2 Hz, 1H), 2.28-2.05 (m, 1H), 1.65-1.49 (m, 4H), 1.49-1.27 (m, 4H).

[0283] Part 3-synthesis of (S)-7-((S)-2-(benzyloxy)-1-cyclopentyl-2-oxoethyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester was charged into a reactor at 15-25°C with (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester (19.45 kg, 83.18 mol, 1.0 equiv) and DCM (234.00 L, 12 V), followed by the addition of DIPEA (21.50 kg, 166.36 mol, 2.0 equiv). The reaction mixture was cooled to -10-0°C under a flow of N2. Tf₂O (31.85 kg, 116.45 mol, 1.4 equiv) was added dropwise to the reaction mixture over 1 h. The reaction mixture was stirred at -10 to 0 °C for 1 h and samples were taken for HPLC analysis. H₂O (19.50 L, 1 V) was added to the reactants at 0 to 5 °C, followed by compound B (1 / 2 tartrate, 27.70 kg, 91.92 mol, 1.1 equiv) and K₃PO₄ (45.90 kg, 216.23 mol, 2.6 equiv).The reactants were heated to 20–25 °C and stirred for 6 h. After 1 h, an aqueous HCl solution (0.5 M, 293.00 L, 15 V) was added to the reactants at 10 ± 5 °C. The mixture was stirred for 30 min and the organic phase was separated. H₂O (195.00 L, 10 V) was added to the organic phase at 10 ± 5 °C. The mixture was stirred for 30 min and the organic phase was separated. The organic phase was concentrated to dryness to give a yellow oily (S)-7-((S)-2-(benzyloxy)-1-cyclopentyl-2-oxoethyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (33.76 kg, 93.2% LCAP, 91.6% yield). Table 14 summarizes the HPLC method used for this synthesis.

[0284] LCMS (ESI+): C26H38N2O4[M+H]+ Calculated value: 443.3; Experimental value: 443.4. H NMR (400 MHz, DMSO-d 6) δ 7.63-7.15 (m, 5H), 5.13 (d, J= 1.3 Hz, 2H), 3.24 (dt, J= 11.5, 5.1 Hz, 1H), 3.19 (d ,J= 8.0 Hz, 1H) , 3.14 (d ,J= 10.4 Hz, 1H) , 3.09 (d ,J= 10.6 Hz, 1H) , 3.04 (d ,J= 10.4 Hz, 1H) , 2.79 (dt,J= 12.8, 5.4 Hz, 1H) , 2.68 (t,J= 7.7 Hz, 1H) , 2.61 (dd,J= 8.8, 3.1 Hz, 1H) , 2.52 (d,J= 1.8 Hz, 1H) , 2.29‑ 2.11 (m, 1H) , 1.84‑1.64 (m, 3H) , 1.60 (t,J= 7.0 Hz, 2H), 1.57-1.44 (m, 5H), 1.39 (s, 10H), 1.16-1.03 (m, 1H).

[0286] Table 14: HPLC and IPC methods for parts 2 and 3:

[0287] Part 4 - Synthesis of (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate specification 92 / 123 pages 104 CN 122270438 A (S)-7-((S)-2-(benzyloxy)-1-cyclopentyl-2-oxoethyl)-2,7-diazaspiro[4.4] Tert-butyl nonane-2-carboxylate (30.1 kg, 68.00 mol, 1.0 equiv) and EtOAc (150.50 L, 5 V). The solution was cooled to 0–5 °C. An EtOAc solution containing HCl (4 M, 150.50 L, 5 V) was added dropwise to the reactants. The reactants were heated to 20–30 °C and stirred at 20–30 °C for 6 h. The reaction was monitored by HPLC, at which point HPLC analysis showed that the reaction was complete. After completion, the reactants were cooled to 0–10 °C, and an aqueous HCl solution (0.5 M, 301.00 L, 10 V) was added to the reactants. The mixture was stirred at 20–30 °C for 20 min, allowed to stand, and the phases were separated. The organic phase was extracted with an aqueous HCl solution (0.5 M, 301.00 L, 10 V) at 0–20 °C. The combined aqueous phases were washed with MTBE (2 x 30 1.00 L, 2 x 10 V). The pH of the aqueous phases was adjusted to 11-12 with an aqueous solution of NaOH (20 wt%) at 5-10 °C. The aqueous phases were extracted with MTBE (2 x 30 1.00 L, 2 x 10 V). The combined organic phases were washed with an aqueous solution of NaCl (26 wt%, 30 1.00 L, 10 V). The organic phases were concentrated to dryness under vacuum to give crude (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)acetate (analytical 20.99 kg, 90.1% yield) as a colorless oil.

[0288] Crude (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate (20.99 kg, 61.29 mol, 1.0 equiv) and IPA (210.00 L, 10 V) were added to the reactor at 20 ± 10 °C. In another reactor, a solution of D-(+)-2,3-dibenzoyl tartaric acid (21.96 kg, 61.29 mol, 1.0 equiv) in IPA solution (210.00 L, 10 V) was prepared. One-fifth of the IPA solution (42.00 L, 2 V) of D-(+)-2,3-dibenzoyl tartaric acid was added dropwise to a (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl acetate solution at 40-45 °C for 1.5 h. Then, seed crystals of (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl acetate (0.21 kg, 0.01 w / w) were added to the reactants at 40-45 °C. The mixture was stirred at 40-45 °C for 1.5 h. After 6 h, the remaining (4 / 5) of the IPA solution containing D-(+)-2,3-dibenzoyl tartaric acid (168.00 L, 8 V) was added dropwise.The solution was added dropwise to the reactants. The mixture was stirred at 40–45 °C for 3 h and then cooled to 0–5 °C. The mixture was stirred at 0–5 °C for another 6 h. The slurry was filtered, and the filter cake was washed with IPA (42.00 L, 2 V) and MTBE (42.00 L, 2 V). The filter cake was dissolved in H₂O (210.00 L, 10 V), and MTBE (420.00 L, 20 V) was added to the solution. The pH of the aqueous phase was adjusted to 11–12 with NaOH aqueous solution (10 wt%). The mixture was stirred for 45 min, then allowed to stand and the phases were separated. The organic phase was washed with NaCl aqueous solution (26 wt%, 210.00 L, 10 V). The organic phase was concentrated to dryness under vacuum to give crude (S)-2-cyclopentyl-2-((S)-2,7-diazaspiro[4.4]non-2-yl)acetate (19.98 kg, 99.96% LCAP, 99.01% ee, 85.8% yield) as a colorless oil. Table 15 summarizes the HPLC method used for this synthesis.

[0289] LCMS (ESI+): C21H30N2O2[M+H]+ Calculated value: 343.2; Experimental value: 343.3. H NMR (400 MHz, CDC13) δ 7.82-7.30 (m, 4H), 5.15 (s, 2H), 3.13 (d, J= 10.6 Hz, 1H), 2.92 (dtd, J= 16.7, 9.4, 5.1 Hz, 3H) , 2.82 (d ,J= 10.8 Hz, 1H) , 2.79‑2.65 (m , 3H) , 2.60 (d ,J= 8.6 Hz , 1H) , 2.27 (dq ,J= 10.4 , 7.8 Hz , 1H) , 1.86‑1.74 (m, 1H) , 1.69 (tdd, J = 10.0, 6.2, 4.2 Hz, 4H), 1.63–1.48 (m, 4H), 1.48–1.38 (m, 1H), 1.20 (s, 2H).

[0291] Table 15: HPLC method for Part 4 IPC method: Specification 93 / 123 pages 105 CN 122270438 A

[0292] Example 11 Alternative synthetic route for (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester Part 1 Synthesis of (R)-2-cyclopentyl-2-hydroxyacetic acid H2SO4 (56.76 kg, 578.78 mol, 2.5 equiv) and H2O (660 mol) were charged into the reactor at 20–25 °C.0.00 L, 20 V) was added, followed by (R)-2-amino-2-cyclopentylacetic acid (33.15 kg, 231.51 mol, 1.0 equiv). The reaction mixture was stirred at 20–25 °C for 30 min to form a homogeneous solution. The solution was filtered through a microporous filter (0.22 µm) into tank A as solution A. NaNO2 (102.45 kg, 1484.88 mol, 6.4 equiv) and H2O (800.00 L, 24.1 V) were added to another reactor at 20–25 °C. The reaction mixture was stirred at 20–25 °C for 30 min and transferred to tank B as solution B. Solutions A and B were mixed in a continuous microreactor. The flow rates were A: 37.7 L / h, B: 27.7 L / h, the temperature was 50 °C, and the retention time in the microreactor was 2 min. The solution was purged with N2 gas for 12 h to push most of the NO2 gas into the exhaust gas absorption reactor. NaCl (198.9 kg, 6 w / w) was added to the reactor and stirred to dissolve the solids. The solution was extracted with MTBE (2 x 994.50 L, 2 x 30 V). The organic phase was concentrated to 2-3 V under reduced pressure (0.08 MPa, 20-30 °C). The MTBE residue was exchanged with n-heptane to 2-3 V under reduced pressure (0.08 MPa, 20-30 °C) until the MTBE residue was less than 3 wt% and KF was less than 0.5 wt%. EtOAc (19.90 L, 0.6 V) was added to the n-heptane residue and the mixture was heated to 45 ± 2 °C. The reaction mixture was stirred at 45 ± 2 °C for at least 8 h and then gradually cooled to 30 °C at a rate of 1.5 °C / h. The slurry was filtered and the filter cake was washed with EtOAc / n-heptane (1:5 v / v, 33.00 L, 1 AV). The wet filter cake was dried under vacuum (0.09 MPa, 45±5 °C) to give (R)-2-cyclopentyl-2-hydroxyacetic acid (13.3 kg, 91.4% LCAP, 44.9% yield) as a grayish-white solid. Table 16 summarizes the HPLC method used for this synthesis.

[0293] LCMS (ESI+): C7H12O3[M-H2O]+ Calculated value: 127.1; Experimental value: 127.3. 1H NMR (400 MHz, DMSO-d 6) δ 3.80 (d, J= 5.9 Hz, 1H), 2.13 (td, J= 7.9, 5.9 Hz, 1H), 1.71-1.50 (m, 4H), 1.50-1.30 (m, 4H).

[0295] Table 16: HPLC and IPC methods used for Part 1:

[0296] Part 2 - Synthesis of (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester: Dimethylacetamide (128.00 L, 5 V), DIPEA (34.44 kg, 266.46 mol, 1.5 equiv), and (R)-2-cyclopentyl-2-hydroxyacetic acid (25.6 kg, 177.57 mol, 1.0 equiv) were charged into a reactor at 20-25°C under a N2 atmosphere. The reaction mixture was cooled to 0-10°C and BnBr (36.45 kg, 213.11 mol, 1.2 equiv) was added dropwise. The reaction mixture was heated to 15-25°C and stirred for 6 h at 15-25°C. The reaction was monitored by GC, at which point GC analysis showed that the reaction was complete. DMAP (13.01 kg, 106.49 mol, 0.6 equiv) was added, and the mixture was stirred at 20–25 °C for 18 h and sampled for GC analysis to ensure that BnBr was not detected. H₂O (256.00 L, 10 V) was added to the solution, and the resulting solution was extracted with MTBE (2 x 128.00 L, 2 x 5 V). The combined organic phases were washed with aqueous citric acid solution (2 x 128.00 L, 2 x 5 V) and H₂O (128.00 L, 5 V), respectively. The organic phases were concentrated to 2–3 V under reduced pressure (0.08 MPa, 20–35 °C). The MTBE residue was exchanged to 2–3 V with DCM (256.00 L, 10 V) under reduced pressure (0.08 MPa, 20–35 °C) until GC analysis showed that the MTBE residue was less than 3% LCAP. A solution of (R)-2-cyclopentyl-2-hydroxyacetic acid benzyl ester in DCM was obtained (44.02 kg analysis, 95.6% LCAP, 105% yield). Table 17 summarizes the HPLC method used in this synthesis according to the specification, page 95 / 123, CN 122270438 A.

[0297] LCMS (ESI+): C14H18O3[M+Na]+ Calculated value: 257.1; Experimental value: 257.2. 1H NMR (400 MHz, DMSO-d 6) δ 7.47-7.27 (m, 5H), 5.38 (d, J= 6.1 Hz, 1H), 5.13 (d, J= 1.2 Hz, 2H), 3.93 (t, J= 6.2 Hz, 1H), 2.28-2.05 (m, 1H), 1.65-1.49 (m, 4H), 1.49-1.27 (m, 4H).

[0299] Table 17: HPLC and IPC methods for sections 1 and 2:

[0300] Example 12 - Synthesis procedure for (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid (compound E) and its sodium salt Section 1 - Synthesis of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl ester at 15±10°C in a reactor by dissolving compound A Ba salt (8.5 kg, 40.88 mol, 1.0 equiv). (Instructions 96 / 123 pages 108 CN 122270438 A in DMF) In a solution of DMF (56.00 L, 4 V), NMI (13.4 kg, 163.22 mol, 4.0 equiv) and compound D (14.00 kg, 40.88 mol, 1.0 equiv) was added dropwise to the solution, followed by the addition of TCFH (13.80 kg, 49.18 mol, 1.2 equiv) dropwise to a solution of DMF (4.00 L, 1 V) over 1 h at 15 ± 10 °C. The reaction mixture was stirred at 15 ± 10 °C for at least 30 min. The reaction was monitored by HPLC, at which point HPLC analysis indicated completion. The reaction mixture was cooled to 5 ± 5 °C and quenched with H2O (28.00 L, 2 V). The mixture was then heated to 20 ± 5 °C and stirred for 10 min. MTBE (280.00 L, 20 V), H₂O (182.00 L, 13 V), and Na₂CO₃ (1.12 kg, 0.08 w / w) were added to the mixture and stirred for 20 min. The organic phase was separated, and the aqueous phase was extracted with MTBE (140.00 L, 10 V). The combined organic phases were washed with an aqueous solution of N₂CO₃ (15 wt%, 2 x 164.60 kg, 2 x 11.76 w / w). The organic phase was concentrated to 4-5 V under reduced pressure (0.08 MPa, 20–40 °C). The process was repeated twice by exchanging MTBE with EtOAc (280.00 L, 20 V) and concentrating to 4-5 V under reduced pressure (0.08 MPa, 20–40 °C). A portion of EtOAc (210.00 L, 15 V) was added to the residue, and the mixture was stirred for 40 min. The solution was cooled to -5±5℃ and maleic acid (2.37 kg, 20.42 mol, 0.5 equiv) was added to the solution in portions. (S)-2-cyclopentyl-2-((S)-7-((2R)Seeds of maleate of benzyl (3R)-3-cyclopropyl-1-methylaziridine-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetate were added to the reactants and stirred for 15 min. A solution of maleic acid (6.17 kg, 1.3 equiv) in EtOAc (280.00 L, 20 V) was added dropwise to the reactants over 7.5 h, and stirring was continued at -5 ± 5 °C for another 7.5 h. The slurry was filtered, and the filter cake was washed with pre-cooled EtOAc (42.00 L, 3 V) at -5 ± 5 °C. The wet filter cake was collected. MTBE (140.00 L, 10 V) and the wet filter cake were added to the reactor at 0 ± 5 °C. Na2CO3 aqueous solution (15 wt%) was added at 0 ± 5 °C until the pH of the aqueous layer was 9–11. The mixture was heated to 25 ± 5 °C and stirred for 30 min. The organic phase was separated, and the aqueous phase was extracted with MTBE (140.00 L, 10 V). The combined organic phases were washed with an aqueous solution of Na₂CO₃ (15% wt%, 140.00 L, 10 V). The organic phase was concentrated to 4–5 V under reduced pressure (0.08 MPa, 20–40 °C). The MTBE was replaced with MeOH (280.00 L, 20 V) by adding MeOH and concentrating to 4–5 V under reduced pressure (0.08 MPa, 20–40 °C), and this process was repeated twice. A portion of MeOH (112.00 L, 8 V) was added to the residue, and the mixture was stirred at 20 ± 10 °C for 30 min. Samples were taken for GC analysis (MTBE residue ≤ 3 wt%). A MeOH solution (128.7 kg solution of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate was obtained, yielding 9.65 kg of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate, 97.8% LCAP, 50.7% yield.

[0301] LCMS (ESI+): C28H39N3O3[M+H]+ Calculated value: 466.3; Experimental value: 466.6.

[0302] ¹H NMR (400 MHz, methanol-d⁴) δ 7.49–7.30 (m, 5H), 6.27 (s, 4H), 5.41–5.20 (m, 2H), 3.87–3.69 (m, 2H), 3.69–3.32 (m, 7H), 2.83 (d⁴, J = 2.4 Hz, 3H)2.55 (dt,J= 12.0, 8.2 Hz, 1H) , 2.45 (t,J= 7.5 Hz, 1H) , 2.27‑ 1.96 (m, 4H) , 1.96‑ 1.84 (m, 1H) , 1.77‑1.63 (m, 1H) , 1.53 (d ,J= 4.8 Hz, 5H) , 1.37‑1.23 (m, 1H) , 0.85‑0.61 (m, 4H) , 0.53 (tq,J= 6.3, 4.3 Hz, 1H).

[0303] Part 2 - Synthesis of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid (compound E) Specification 97 / 123 pages 109 CN 122270438 A A MeOH solution (6.05 kg analysis, 13.00 mol, 1.0 equiv) of benzyl (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl ester and TEA (4.00 kg, 39.00 mol, 3.0 equiv) were added to the reactor. The mixture was cooled to 0±5℃ and PdCl2 (60.50 g, 1% w / w) was added to the reactants. The reaction mixture was stirred at 0±5℃ and 0–0.15 MPa H2 pressure for at least 3 h. Samples were taken for HPLC analysis. The reaction mixture was filtered through diatomaceous earth and the filter cake was washed with MeOH (42.35 L, 7 V). The filtrate was concentrated to 8–10 V under reduced pressure (20–30℃, 0.08 MPa). The solvent was exchanged twice to 8–10 V with THF (60.50 L, 10 V) under reduced pressure (20–30℃, 0.08 MPa). THF (60.50 L, 10 V) and TEA (2.00 kg, 19.76 mol, 1.5 equiv) were added to the residue. The resulting solution was concentrated to 8–10 V under reduced pressure (20–30 °C, 0.08 MPa), and THF (18.15 L, 3 V) was added to the residue. The solution was stirred at 20 ± 10 °C for at least 30 min to form a slurry. MTBE (60.50 L, 10 V) was added to the slurry at 20 ± 10 °C for 1.2 h. The mixture was concentrated to 10–13 V under reduced pressure (20–30 °C, 0.08 MPa). The solution was then concentrated using MTBE by dropwise addition over 1 h followed by reduced pressure (20–30 °C, 0.08 MPa).(48.40 L, 8 V) The solvent was exchanged to 10–13 V for 6 cycles. MTBE (48.40 L, 8 V) was added dropwise to the suspension at 20 ± 10 °C for 1 h and samples were taken for analysis (THF residue ≤ 7% w / w). The slurry was filtered and the filter cake (18.15 L, 3 V) was washed. The filter cake was dried under vacuum (30 ± 5 °C, 0.08 MPa) for at least 8 h until LOD ≤ 10% w / w. Compound E was obtained as a grayish-white solid (4.13 kg, 95.9% LCAP, 90.2% w / w analysis, 76.4% yield).

[0304] LCMS (ESI+): C21H33N3O3[M+H]+ Calculated value: 376.3; Experimental value: 376.4. 1H NMR (400 MHz, methanol-d 4) δ 3.87-3.76 (m, 1H), 3.73-3.63 (m, 1H), 3.63-3.38 (m, 7H), 2.44-2.25 (m, 5H), 2.19 (dq ,J= 13.1, 6.7, 6.2 Hz, 1H), 2.14- 1.95 (m, 3H), 1.90 (qt,J= 6.9, 2.9 Hz, 1H), 1.82-1.51 (m, 7H), 1.46 (dt,J= 9.0, 7.0 Hz, 1H) , 0.73-0.56 (m, 1H), 0.46 (dddd, J = 17.8, 7.5, 5.7, 4.3 Hz, 3H), 0.28 (dtt, J = 11.5, 6.2, 2.7 Hz, 1H).

[0306] The sodium salt of compound E was prepared by adding a MeOH solution (1.0 equiv) of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate and TEA (3.0 equiv) to the reactor. The mixture was cooled to 0±5°C and PdCl2 (1% w / w) was added to the reactants. The reaction mixture was stirred at 0±5℃ and 0-0.15 MPa H2 pressure for at least 3 h. Samples were taken for HPLC analysis. The reaction mixture was filtered through diatomaceous earth and the filter cake was washed with MeOH (7 V). The filtrate was concentrated to 3-4 V under reduced pressure (20-30℃, 0.08 MPa). MeOH (7 V) was added to the residue and the solution was stirred at rt for 10 min. NaOMe (1.03 g / L) was added at rt.Add 30 wt% MeOH solution (equiv) to the solution and stir the resulting mixture at rt for at least 3 h. Concentrate the reaction solution to 3–4 V under reduced pressure (20–30 °C, 0.08 MPa) to obtain solution A. Charge MTBE (60 V) into another reactor and cool the reactor to -5–5 °C. Add seed crystals (2 wt%) of compound E Na salt to the MTBE solution and stir the suspension for 30 min. After 5 h at -5–5 °C, add solution A of compound E to the MTBE and stir the suspension for 2 h. Filter the slurry and wash the wet filter cake with MTBE (2 V) as per the specification (page 98 / 123, 110 CN 122270438 A). Dry the wet filter cake under vacuum at 30–40 °C for 5 h to obtain compound E Na salt. Table 18 summarizes the HPLC method used for this synthesis.

[0307] LCMS (ESI+): C21H33N3O3[M+H]+ Calculated value: 376.26; Experimental value: 376.28.

[0308] 1H NMR (400 MHz, methanol-d4) δ 3.70-3.65 (m, 1H), 3.57-3.35 (m, 3H), 3.01-2.79 (m, 4H), 2.39 (s, 3H), 2.34 (t, J = 7.2 Hz, 1H), 2.29-2.16 (m, 1H), 2.10-1.70 (m, 6H), 1.70-1.47 (m, 5H), 1.42 (p, J = 7.2 Hz, 2H), 0.69-0.21 (m, 5H).

[0309] Table 18: HPLC and IPC methods for parts 1 and 2:

[0310] Example 13 Alternative synthetic route for benzyl (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4,4]non-2-yl)acetate The reactor was charged with MTBE (90.50 mL, 5 V), compound D (18.10 g, 52.85 mmol, 1.0 equiv), compound A (12.84 g, 55.49 mmol, 1.05 equiv) and 1,5,7-triazabicyclo[4,4,0]dec-5-ene (TBD) (2.21 g, 15.86 mmol, 0.3 equiv). The reaction mixture was cooled to -5 to 0 °C. The reaction mixture was stirred at -5 to 0°C for 24 hours, at which point HPLC analysis indicated the reaction was complete. Then, 181.00 mL of Na₂CO₃ aqueous solution (5 wt%) was added.V) Quench the reaction and stir the mixture at 15–25 °C for at least 10 min. Separate the phases and wash the organic phase with a Na₂CO₃ aqueous solution (5 wt%, 181.00 mL, 10 V). Concentrate the organic phase to dryness (T ≤ 40 °C). Dissolve the residue in EtOAc (145.00 mL, 8 V) and cool the solution to -10–0 °C. Maleic acid (3.07 g, 26.42 mmol, 0.5 equiv) was added to the solution, followed by the addition of seed crystals of maleate of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetate (181.00 mg, 1 wt%) at -10-0 °C. The mixture was stirred for at least 10 min, followed by the dropwise addition of maleic acid to a solution of MTBE / EtOAc (v / v = 3:2, 181.00 mL, 10 V) over 3 h. The suspension was stirred at -10-0 °C for 12 h. The slurry was filtered and the filter cake was washed with pre-cooled MTBE / EtOAc (v / v = 1:2, 54.30 mL, 3 V). The filter cake was transferred to a reactor where MTBE (181.00 mL, 10 V) was cooled to -5 to 0 °C. Na₂CO₃ aqueous solution (15 wt%, 181.00 mL, 10 V) was added to the solution to adjust the pH to 8–10. The mixture was stirred at 15–25 °C for at least 10 min. The phases were separated, and the aqueous phase was extracted with MTBE (90.50 mL, 5 V). The organic phase was washed with Na₂CO₃ aqueous solution (5 wt%, 181.00 mL, 10 V). The organic phase was concentrated to dryness to give maleate of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetate (19.3 g, 97.2% LCAP, 92.7% QNMR analysis).

[0311] Benzyl benzyl acetate of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4,4]non-2-yl)acetate was charged into a reactor with MTBE (5 V), compound D (1.0 equiv), compound A (1.05 equiv), and 1,5,7-triazabicyclo[4,4,0]dec-5-ene (TBD) (0.3 equiv). The reaction mixture was cooled to -5 to 0 °C. The reaction mixture was then subjected to a reaction...The mixture was stirred at -5 to 0°C for 24 hours, at which point HPLC analysis indicated the reaction was complete. Na₂CO₃ aqueous solution (5 wt%, 10 V) was added to quench the reaction, and the mixture was stirred at 15 to 25°C for at least 10 min. The phases were separated, and the organic phase was washed with Na₂CO₃ aqueous solution (5 wt%, 10 V). The organic phase was concentrated to dryness (T ≤ 40°C). Benzenesulfonic acid (2.1 equiv) was dissolved in THF (4.53 V) and reserved for later use. The concentrated (S)-2-cyclopentyl-2-((S)-7-((2R, 3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)benzyl acetate residue was diluted with acetone (10 V). The solution was cooled to 7 ± 3°C. A pre-prepared benzenesulfonic acid solution in THF (3.8 / 21) was added fractionally to (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl acetate, followed by the addition of 0.5% w / w seed crystals of the benzenesulfonate of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridin-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl acetate. The mixture was stirred at 7 ± 3 °C for 1 h, followed by the addition of another fraction of the benzenesulfonic acid THF solution (15.2 / 21). The resulting mixture was stirred at 7 ± 3 °C for 16 h. Next, the remaining benzenesulfonic acid THF solution (0.5 / 21) was added to the mixture and stirring continued for 6 h. The slurry was filtered and the wet filter cake was washed with THF / acetone (v / v = 1:2, 3 V). The wet filter cake was added to MTBE (10 V) and Na2CO3 aqueous solution (15% w / w) was added at 0 ± 5 °C to adjust the pH to 7.5–9.0. H2O (5 V) was added to the mixture and the biphasic solution was stirred for 0.5 h, after which the organic phase was separated. The aqueous phase was washed with MTBE (5 V). The organic phases were combined and washed with Na2CO3 aqueous solution (5% w / w, 10 V). The organic phase was concentrated to 4–5 V under vacuum at NMT 40 °C. The concentrated residue was diluted with acetone (10 V) and the resulting mixture was concentrated to 4–5 V under vacuum at NMT 40 °C. Solvent washing with acetone was repeated once. A solution of (S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)acetic acid benzyl ester in acetone was used in the next step. Table 19 summarizes the HPLC method used for this synthesis.

[0312] LCMS (ESI+): C28H39N3O3[M+H]+ Calculated value: 466.3; Experimental value: 466.6.

[0313] ¹H NMR (400 MHz, methanol-d⁴) δ 7.49–7.30 (m, 5H), 6.27 (s, 4H), 5.41–5.20 (m, 2H), 3.87–3.69 (m, 2H), 3.69–3.32 (m, 7H), 2.83 (d, J = 2.4 Hz, 3H), 2.55 (dt, J = 12.0, 8.2 Hz, 1H), 2.45 (t, J = 7.5 Hz, 1H), 2.27–1.96 (m, 4H), 1.96– (See specification 100 / 123 pages 112 CN 122270438 A) 1.84 (m, 1H) , 1.77‑1.63 (m, 1H) , 1.53 (d ,J= 4.8 Hz, 5H) , 1.37‑1.23 (m, 1H) , 0.85‑0.61 (m, 4H) , 0.53 (tq,J= 6.3, 4.3 Hz, 1H).

[0314] Table 19: HPLC method for benzenesulfonate formation IPC method:

[0315] Example 14 - Synthesis of (S)-3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-3-yl)-2,2-dimethylprop-1-ol (compound H) Part 1 - Synthesis of (S)-5-(2-(1-methoxyethyl)pyridin-3-yl)-2,2-dimethyl-5-oxovalerate Instructions 101 / 123 pages 113 CN 122270438 A Under nitrogen protection and with stirring, THF (230.5 kg, 2 V) and i-PrMgCl·LiCl (465.0 kg, 1.3 mol / L, 1.03 mol / L) were charged into reactor A. The reactor was cooled to -20±5℃. A solution of 3-bromo-2-[(1S)-1-methoxyethyl]pyridine (129.7 kg, 600.2 mol, 1.0 equiv) in THF (350.4 kg, 3 V) was added dropwise (21 kg / min) to the reactor while maintaining the internal temperature at -20±5℃. The reaction mixture was stirred at -20±5℃ for 2 h and sampled for IPC. THF (577.4 kg, 5 V) and 2,2-dimethylglutaric anhydride (89.2 kg, 627.5 mol, 1.05 equiv) were added to reactor B with stirring at -10±5℃. At -10±5℃ (3.9The solution from reactor A was added dropwise to reactor B at a rate of kg / min. The reaction mixture was sampled for HPLC analysis. A solution of 2,2-dimethylglutaric anhydride (0.1 equiv) in THF (0.3 V) was added to the reaction mixture, and the mixture was stirred at -10 ± 5 °C for 5 h. The reaction mixture was sampled for HPLC analysis. The reaction mixture was quenched by adding H₂O (3 V) at -5 ± 5 °C. The mixture temperature was adjusted to 20 ± 5 °C, and the mixture was stirred for at least 0.5 h. n-Heptane (3 V) and 4% NaOH aqueous solution (3 V) were added to the organic phase. After stirring for 0.5 h, the aqueous phase was separated. The combined aqueous layers were adjusted to pH = 7.5 ± 0.3 with KHSO₄ at 20 ± 5 °C. The aqueous phase was washed with n-heptane (5 V x 1, 2 V x 1). The aqueous phase was filtered, and the filtrate was adjusted to pH = 5.7–7.0 with KHSO₄. Seed crystals of (S)-5-(2-(1-methoxyethyl)pyridin-3-yl)-2,2-dimethyl-5-oxovaleric acid (14.76% w / w) were added to the solution. The solution was acidified to pH 5.5 ± 0.2 with KHSO4. The slurry was stirred at 15–25 °C for at least 5 h and filtered. A filter cake of the product, appearing as a pale yellow solid, was obtained.

[0316] LCMS (ESI+): C15H22NO4(M+H) Calculated value: 280.1; Experimental value: 280.1 1H NMR (400 MHz, DMSO-d 6, 25℃) δ 12.2 (s, 1H), 8.59 (dd, J= 4.8, 5.2 Hz, 1H), 7.89 (dd, J= 7.6, 8.0 Hz, 1H), 7.40-7.37 (m, 1H), 4.58-4.53 (m, 1H), 3.11 (s, 3H), 2.81-2.77 (m, 2H), 1.82-1.78 (m, 2H), 1 .42 (d, J = 6.4 Hz, 3H), 1.12 (s, 6H).

[0317] Partial 2-synthesis of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropionic acid was introduced into the reactor with H2O (522 L, 2.5 V). H2SO4 (150.7 kg, 1537 mol, 2.0 equiv) was added dropwise to the reactor at 20±20 °C. (S)-5-(2-(1-methoxyethyl)pyridin-3-yl)-2,2-dimethyl-5-oxovalerate (215.6 kg, 771.8 mol, 1.0 equiv) was added dropwise to the reactor at 20±10 °C.189.5 kg (847.9 mol, 1.1 equiv) and 4-bromophenylhydrazine hydrochloride (189.5 kg, 847.9 mol, 1.1 equiv) were added to the reactor. The reaction mixture was heated to 70–75 °C and stirred for at least 0.5 h. The reaction mixture was then heated at 95 ± 5 °C for 32 h. The mixture was sampled for HPLC analysis. After completion, the reaction mixture was cooled to 60–65 °C. Water (1185.8 kg, 5.5 V) and 30% NaOH aqueous solution (189.3 kg, 1.9 equiv) were added, and the mixture was stirred at 60–65 °C for 5 h. The mixture was cooled to 25 ± 5 °C and stirred for 2 h. The mixture was filtered and washed with H2O (800 L, 3 V). The wet filter cake was added to H2O (1022.5 kg) and H2SO4 (53.9 kg) at 25±5℃. The mixture was heated to 80-85℃ and stirred for 5 h. Then the mixture was cooled to 25-30℃ and stirred for 16 h. The slurry was filtered and washed with H2O (1020 L, 3 V). The filter cake was dried at 50±5℃ for 36 h to obtain a product in the form of a yellow solid.

[0318] LCMS (ESI+): C21H24BrN2O3(M+H) Calculated value: 433.1; Experimental value: 433.1 1H NMR (400 MHz, DMSO-d 6, 25℃) δ 12.1 (s, 1H), 11.4 (s, 1H), 8.70 (s, 1H), 7.82–7.77 (m, 2H), 7.45–7.20 (m, 3H), 4.19 (d, J = 5.6 Hz 1H), 2.93 (s, 4H), 2.78 (s, 1H), 1.36 (d, J = 4.0 Hz, 3H), 0.88 (d, J = 5.2 Hz, 6 H).

[0319] A portion of methyl 3-(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropionate was added to the reactor along with MeOH (5.45 L, 5 V) and (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropionate·1 / 2H2SO4 (1.09 kg, 2.27 mol, 1.0 equiv). Then, SOCl2 (331 g, 2.78 mol, 1.2 equiv) was added at 50–60 °C. The reaction mixture was stirred at 65 °C for 12 h.After completion, the reaction mixture was cooled to 10°C. The pH of the mixture was adjusted to 7-8 with 5% Na₂CO₃ at 10-15°C. The mixture was filtered and the filter cake was washed with H₂O (2.2 L, 2 V). The wet filter cake was wet-milled with H₂O (5.4 L, 5 V) at 25°C for 6 h. The mixture was filtered and the filter cake was washed with H₂O (2.2 L, 2 V). The filter cake was dried in a forced-air oven (N₂, 45-50°C) for 24 h to obtain the product.

[0320] LCMS (ESI+): C22H26BrN2O3(M+H+) Calculated value: 447.1; Experimental value: 446.9 1H NMR (300 MHz, DMSO-d6) δ 11.41 (s, 1H), 8.72 (dd, J= 4.7, 1.8 Hz, 1H), 7.81 (dd, J= 7.7, 1.8 Hz, 1H), 7.65 (d, J= 1.9 Hz, 1H), 7.47 (dd, J= 7.8, 4.7 Hz, 1H), 7.32 (d, J= 8.6 Hz, 1H), 7.22 (dd, J= 8.6, 1.9 Hz, 1H), 4.23 (q,J= 6.3 Hz, 1H), 3.32 (s, 9H), 2.95 (s, 4H), 1.39 (d,J= 6.3 Hz, 3H), 0.94 (s, 6H).

[0321] Part 4 Synthesis of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionate methyl ester Specification 103 / 123 pages 115 CN 122270438 A DMF (1163.1 kg, 5 V), (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropionate methyl ester (239.04 kg, 536.7 mol, 1.0 equiv), and Cs2CO3 (245.94, 754.8 mol, 1.4 equiv) were charged into the reactor. The reactor was flushed with DMF (454.6 kg, 2 V). The reaction mixture was cooled to 15–20 °C and 2,2,2-trifluoroethyl trifluoromethanesulfonic acid (170 kg, 732.4 mol, 1.35 equiv) was added. The reaction mixture was stirred at 15–20 °C for 16 h. A sample of the reaction mixture was taken for HPLC analysis. After completion, the mixture was subjected to HPLC analysis at 15–20 °C with AcOH (64.4 kg, 2.0 mol).The reaction was quenched with methyl methyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionate in diastereomer form (1.4:1). The resulting mixture was extracted twice with MTBE (5 V, 2 V). The combined organic layers were concentrated to 3-4 V at 35-45 °C under reduced pressure. The resulting EtOH solution of methyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionate in diastereomer form was used directly in the next step.

[0322] LCMS (ESI+): C24H27BrF3N2O3(M+H+) Calculated value: 529.1; Experimental value: 529.9 1H NMR (300 MHz, DMSO-d6) δ 8.77 (dt, J = 4.8, 1.4 Hz, 1H), 7.86–7.75 (m, 1H), 7.73–7.62 (m, 2H), 7.55 (ddd, J = 7.8, 6.0, 4.7 Hz, 1H), 7.40 (ddd, J = 8.8, 4.0, 1.9 Hz, 1H), 5.37 (dd, J = 16.5, 8.6 Hz, 1H), 4.82 (dd,J= 17.9, 9.0 Hz, 1H) , 4.46 (dd,J= 16.4, 9.5 Hz, 0H) , 3.93 (dt,J= 18.4, 6.2 Hz, 1H) , 3.66 ‑ 3 .56 (m , 2H) , 3 .46 (d ,J= 13 .2 Hz , 3H) , 3 .06 (s , 1H) , 2.94 (s , 2H) , 1 .84 ‑ 1.70 (m, 2H) , 1.32 (dd,J= 37.8, 6.2 Hz, 3H) , 1.05 ‑ 0.87 (m, 7H).

[0323] A mixture of the transisomers of the 5-synthetic compound G-(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol was charged into a reactor at 15-20°C under N2.A solution of methyl 2,2-dimethylpropionate (1.0 equiv, 5 V) in EtOH was prepared. CaCl2 (60.04 kg, 541.0 mol, 1.0 equiv) was added in portions to the mixture at 20–30 °C, followed by NaBH4 (51.04 kg, 1349.2 mol, 2.5 equiv) at 20–30 °C. The reaction mixture was stirred at 20–30 °C for 14 h. Samples of the reaction mixture were taken for HPLC analysis. The reaction was quenched by dropwise addition of HCl (3 M) until pH = 1–2. An aqueous solution of NaOH (30% w / w) was added to the mixture to adjust the pH to 4–4.5. The resulting mixture was concentrated to 3–5 V. The concentrated mixture was further diluted with MTBE (891.2 kg, 5 V) and H₂O (1197.5 kg, 5 V). The organic phase was separated. The organic layer was washed sequentially with H₂O (718.5 kg, 3 V), NaOH aqueous solution (484.0 kg, 2 V), and H₂O (718.5 kg, 3 V). The organic layer was concentrated to 3–5 V at 45–60 °C. The concentrated mixture was diluted with IPA (756.3 kg, 4 V) and further concentrated under reduced pressure at 45–60 °C to approximately 3–4 V. The previous unit operation was repeated until the MTBE content in the IPA solution was less than 2% w / w. The resulting IPA solution (247.6 kg, 98.8% a / a, 92% separation yield) of the compound G mixture in diastereomer form, purified by column purification or by chemical resolution.

[0324] Part 6 - Purification of the compound G mixture under the following conditions: The compound G mixture (19.0 kg) was treated with SiO2 (30.0 kg) for column chromatography. Silica gel was loaded onto a column chromatography column (240 kg SiO2, n-heptane / EtOAc = 1 / 0 to 0 / 1) to obtain a pure fraction (about 1000 L) containing compound G. The fraction was concentrated to about 25 L. n-Heptane (100 L) was added to the concentrated residue and the resulting mixture was concentrated to about 25 L. The n-heptane washing was repeated once. The residue was diluted with n-heptane (100 L) and the mixture was stirred at 15-20°C for 12 h. The slurry was filtered and the wet filter cake was dried in an oven under vacuum to obtain compound G (18.0 kg, 98.9% a / a) as a white solid.

[0325] Condition 2: An IPA solution of the compound G mixture was added to the reactor by CSA resolution, and IPA (1314.0 kg, 1.2%) was added to the reactor.Next, an IPA solution of the compound G mixture (analytical weight 247.6 kg, 495.8 mol, 1.0 equiv) was pumped through a continuous reactor and a quenching reactor (continuous reactor: 220±10℃, quenching reactor: 0–20℃, back pressure: 4.00±0.20 MPa) at a flow rate of 80.0 L / h. The equilibrium IPA solution of compound G was added to the reactor while stirring at 20±10℃. The mixture was sampled for HPLC analysis (standard: diastereomer ratio ≥1.77). D-camphor-10-sulfonic acid (CSA, 150.0 kg, 645.7 mol, 1.3 equiv) was added to the reaction mixture, followed by IPA (21.5 kg, 0.1 V) to flush the feed port. The reaction mixture was stirred at 25±5℃ for 2 h and then at 53±3℃ for 2 h. Compound G CSA seed crystals (0.1% w / w) were added to the reaction mixture at 43±3 °C and stirred for 2 h at 43±3 °C. The solution was further cooled to -6±3 °C and stirred for 4 h at the same temperature. The resulting suspension was filtered and washed with IPA (212.0 kg, 1.0 V). To further control for transisomer impurities of compound G, the wet filter cake was resuspended in IPA (10 V) and stirred at 67–70 °C for 3–4 h. The mixture was cooled to 5 °C and stirred for 2 h at 5 °C. The resulting wet filter cake (analytical weight 200.0 kg, 1.0 equiv) was suspended in MTBE (3.5 V) at 20±5 °C, followed by the addition of H2O (4 V). The pH was adjusted to 8–10 by slowly adding an aqueous solution of NaOH (30% w / w). The mixture was stirred at 20±5 °C for 0.5 h. The organic phase was separated and washed with H₂O (3 x 3 V). The organic layer was concentrated to 1–2 V. The concentrated mixture was diluted with n-heptane (3 V) and further concentrated under reduced pressure to approximately 2–3 V. Solvent exchange with n-heptane was repeated twice. The concentrated mixture was diluted with n-heptane (3 V) and stirred at 55 ± 5 °C for 3 h. The mixture was cooled to 20 ± 5 °C and stirred at 20 ± 5 °C for 1 h. The slurry was filtered and washed with n-heptane (1.2 V). The wet filter cake was dried at 45–55 °C to give a product as a grayish-white solid (143.6 kg, 99.05% a / a, 58% yield). To further control the trans-isomer impurities of compound G, the solid was resuspended in THF / n-heptane (v / v = 1:4, 5 V) and stirred at 67–70 °C for 3–4 h. Cool the mixture to 5°C and stir it at 5°C for 2 seconds.h. The filtered slurry was washed with n-heptane (1.5 V). The wet filter cake was dried at 45–55 °C to give compound G as a grayish-white solid. To recover compound G, the mother liquor from the resolution process was loaded into the reactor. The solution was concentrated to 4–5 V under vacuum at NMT 50 °C. A 30% w / w NaOH aqueous solution was added to the concentrated residue at 15–30 °C to adjust the pH to 8–10. The mixture was concentrated to 0.8–1.2 V under vacuum at NMT 50 °C. MTBE (5 V) and H₂O (5 V) were added to the residue. The two-phase system was stirred at 25 ± 5 °C for 0.5 h. The organic phase was separated and washed with H₂O (5 V). The organic phase was then concentrated to 1.2–1.6 V at NMT 50 °C. The concentrated residue was diluted with IPA (1.6 V) and then concentrated to 1.2–1.6 V at NMT 50 °C. Solvent exchange with IPA was repeated once, and IPA (3 V) was added to the concentrated residue. The IPA solution of the compound G mixture was subjected to equilibration and CSA salt formation again. The IPA solution of the compound G mixture was pumped through a continuous reactor and a quenching reactor (continuous reactor: 220 ± 10 °C; quenching reactor: 0–20 °C; back pressure: 4.00 ± 0.20 MPa) at a pump flow rate of 80.0 L / h. The equilibrated IPA solution of compound G was loaded into the reactor while agitation was maintained at 20 ± 10 °C. The mixture was sampled for HPLC analysis (standard: diastereomer ratio ≥ 1.77). D-camphor-10-sulfonic acid (1.3 equiv) was added to the reaction mixture, followed by IPA (0.1 V) to flush the feed port. The reaction mixture was stirred at 25±5 °C for 2 h, followed by stirring at 53±3 °C for 2 h. CSA seed crystals (0.1% w / w) of compound G were added to the reaction mixture at 43±3 °C, and the mixture was stirred at 43±3 °C for 2 h. The solution was further cooled to -6±3 °C and stirred at that temperature for 4 h. The resulting suspension was filtered and washed with IPA (1.0 V). The resulting wet filter cake (1.0 equiv) was suspended in MTBE (3.5 V) at 20±5 °C, followed by the addition of H2O (4 V). The pH was adjusted to pH 8–10 by slowly adding an aqueous solution of NaOH (30% w / w). The mixture was stirred at 20±5 °C for 0.5 h. The organic phase was separated and washed with H2O (3 x 3 V). The organic layer was concentrated to 1–2 V. The concentrated mixture was diluted with n-heptane (3 V) and further concentrated under reduced pressure to approximately 2–3 V. Solvent exchange with n-heptane was repeated twice. The concentrated mixture was diluted with n-heptane (3 V) and stirred at 55 ± 5 °C for 3 minutes.h. The mixture was cooled to 20±5℃ and stirred at 20±5℃ for 1 h. The slurry was filtered and washed with n-heptane (1.2 V). The wet filter cake was dried at 45-80℃ to obtain compound G, a grayish-white solid of pure transisomer.

[0326] Condition 3: Xylene (3 V) was added to a toluene solution of the compound G mixture by resolution with methanesulfonic acid (MsOH). The mixture was concentrated to 2.5-3.5 V. The residue was stirred at 137±5℃ for 24 h. The mixture was cooled to 60-70℃ and sampled for HPLC analysis. The residue was diluted with 2-MeTHF (5 V) at 20±5℃. The solution was washed twice with aqueous HCl (0.2 M, 3 V). The organic phase was separated and extracted twice with aqueous HCl (3 M, 3 V). The combined aqueous phase was washed with MTBE (3 V). The aqueous phase was adjusted to pH 8-10 with 30% NaOH aqueous solution. Extract the aqueous phase with IPAc (5 V). Extract the organic phase and wash with H2O (3 V). Concentrate the organic phase to 2–3 V under reduced pressure. Repeat solvent exchange with IPAc twice. Dilute the concentrated residue with IPAc (5 V). Add methanesulfonic acid (MSA, 0.35 equiv) to the IPAc solution at 20 ± 10 °C. Stir the mixture at 30 ± 5 °C for 4 h. Filter the slurry and wash the filter cake with IPAc (2 V). Collect and recover the filter cake containing the unwanted transisomer of compound G in the form of methanesulfonate. Add 5% NaHCO3 aqueous solution (3 V) to the filtrate, ensuring the pH of the aqueous phase is 8. Add H2O (3 V) to the two-phase system. Separate the organic phase and concentrate it to 1.8–2.3 V under vacuum at NMT 45 °C. Dilute the concentrated residue with MeOH (7 V). Concentrate the resulting solution to 1.8–2.3 V under vacuum at NMT 45 °C. MSA (1.0 equiv) was added to the MeOH solution at ±10℃ on pages 106 / 123 of the specification (CN 122270438 A). The mixture was stirred at 40℃ for 0.5 h until all solids were dissolved. H2O (2.4 V) was added to the solution after 2 h, followed by the addition of compound G seed crystals (10% w / w). The mixture was stirred at 40℃ for 4 h, followed by the dropwise addition of H2O (1.8 V). Another portion of H2O (6 V) was added dropwise and the mixture was stirred for 1 h. The mixture was cooled to 20℃ and the slurry was stirred for 4 h. The slurry was filtered and the wet filter cake was washed with MeOH / H2O (v / v=1:3.5, 1.8 V). The wet filter cake was dried in an oven at 50–55℃ for 16 h to obtain compound G as a white solid.Collect compound G and concentrate the filtrate under vacuum. Combine the residue with the wet filter cake containing the undesired transisomer of compound G in the form of methanesulfonate. Add an aqueous solution of NaOH (30% w / w) to adjust the pH to 8–10. Add MTBE (5 V) to the reaction mixture. Stir the mixture at 20 ± 5 °C for 1 h. Separate the organic phase and concentrate to 1.5–3 V. Dilute the concentrated residue with MTBE (5 V). Concentrate the resulting mixture to 1.5–3 V. Repeat the solvent exchange with MTBE twice. Concentrate the compound G MTBE solution to 1.5–2.5 V and add xylene (3 V) to the residue. Stir the mixture at 138–143 °C for 24 h, then cool it to 60 °C. Add IPA (6.3 V) to the mixture and concentrate the resulting solution to 2–3 V under vacuum. Repeat the solvent exchange with IPA twice. D-camphor-10-sulfonic acid (1.3 equiv) was added to the IPA solution at 15 ± 5 °C. The mixture was cooled to -5 to 0 °C and stirred at -5 to 0 °C for 14 h. The slurry was filtered and washed with IPA (2 V). The wet filter cake was dried at 45 to 80 °C to give compound G as a grayish-white solid.

[0327] LCMS (ESI+): C23H27BrF3N2O2(M+H+) Calculated value: 499.1; Experimental value: 499.1 1H NMR (400 MHz, DMSO-d6) δ 8.75 (dd, J = 4.8, 5.2 Hz, 1H), 7.98 (d, J = 1.6 Hz, 1H), 7.78–7.76 (m, 1H), 7.66–7.63 (m, 1H), 7.54–7.53 (m, 1H), 7.40–7.38 (m, 1H), 5.39–5.30 (m, 1H), 4.60– 4.56 (m, 1H), 4.51–4.42 (m, 1H), 3.98–3.93 (m, 1H), 3.33 (s, 2H), 3.03–2.94 (m, 3H), 2.60–2.50 (m, 1H), 2.29–2.26 (m, 1H), 1.36 (d, J = 6.4 Hz, 2H), 0.60 (s, 6H).

[0328] Partial 7a-(S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane)The two-step synthesis of heteroborane-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol involved dissolving compound G (200.0 kg, 400.5 mol, 1.0 equiv) in THF (500.0 L, 2.5 V) and heptane (500.0 L, 2.5 V) at 20 ± 5 °C. HBPin (53.8 kg, 420.4 mol, 1.05 equiv) was added dropwise to the reaction mixture at 20 ± 5 °C, and the mixture was stirred at 30–35 °C for 2 h. The reaction mixture was sampled for 1H NMR to indicate complete consumption of compound G. The reaction mixture was then cooled to 20 °C. B2Pin2 (119.0 kg, 468.6 mol, 1.2 equiv), dtbpy (2.1 kg, 8.01 mol, 0.02 equiv), and [Ir(OMe)(COD)]2 (1.3 kg, 2.0 mol, 0.005 equiv) were added sequentially to the reaction mixture. The resulting mixture was stirred at 30–35 °C for 12 h. Samples were taken for HPLC analysis. The reaction was quenched by charging H2O (0.1 V) and THF (0.5 V) at 0–5 °C, followed by charging MTBE (4 V) and H2O (4 V). The mixture was filtered and the filter cake was washed with MTBE (3 V). The filtrate was separated and the organic layer was washed with H2O (3 V). The organic layer was concentrated to 1–2 V at 40–45 °C. The concentrated mixture was diluted with DCM (4 V) and further concentrated to 1–2 V. Repeat the previous unit operation twice. The resulting DCM solution of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4, 5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol was used directly in the next step (479 kg, 86% analytical yield, 43.1% w / w, 85.1% a / a).

[0329] Partial one-step synthesis of 7b-(S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol was carried out at 25°C by charging a reactor with n-heptane (4 V) and dtbpy (1.6 mol%), followed by the addition of compound G (1.0 mol%).[Ir(OMe)(COD)]2 (0.8 mol%) was added to the suspension under a N2 atmosphere. The reaction mixture was stirred under N2 gas until all solids were dissolved. The reaction mixture was then stirred at 55°C for 2 h. After completion, the reaction solution was washed twice with H2O (3 V). The organic phase was concentrated to dryness and used for the next step.

[0330] LCMS (ESI+): C23H27BBrF3N2O4 (M-2,3-dimethylbutane) Calculated value: 544.1; Experimental value: 544.0 1H NMR (300 MHz, chloroform-d) δ 9.11 (d, J = 1.7 Hz, 1H), 8.10 (t, J = 1.5 Hz, 1H), 7.92 (d, J = 1.9 Hz, 1H), 7.40 (dd, J = 8.7, 1.9 Hz, 1H), 7.29 (s, 1H), 4.73 (s, 1H), 4.47 (q, J = 8.6 Hz, 2H), 4.03 (q, J = 6.2 Hz, 1H), 3.82-3.72 (m, 1H), 3.58-3.45 (m, 2H), 3.04 (s, 3H), 2.74 (d, J = 14.1 Hz, 1H), 2.23 (d, J = 14.1 Hz, 1H) , 1.91 - 1.83 (m, 1H), 1.46 (d, J = 6.2 Hz, 4H), 1.36 (s, 12H), 1.28 (d, J = 5.2 Hz, 35H), 0.95 - 0.85 (m, 5H), 0.76 (d , J = 10.4 Hz, 6H).

[0331] Partial 8-Synthetic compound H-(S)-3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol Specification 108 / 123 pages 120 CN 122270438 A Condition 1: TMP as base DCM (1173 L, 30 V), 2,2,6,6-tetramethylpiperidine (35.3 kg, 250.0 mol, 4.0 equiv), 1-cyclopropylpiperazine (19.7 kg, 156.1 equiv) were added to the reactor under stirring at 15-20°C.14.2 kg (2.5 equiv) and Cu(OAc)₂ (14.2 kg, 78.2 mol, 1.25 equiv). 21% O₂ was bubbled under the surface for 1 h at 20–25 °C. DCM (39.1 kg analytical weight, 62.5 mol, 1.0 equiv) containing (S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol was added to the reaction mixture at 10–20 °C. The reaction mixture was heated at 25–35 °C, during which 21% O₂ was bubbled under the surface, and the mixture was stirred under these conditions for 16 h. Samples were taken for HPLC analysis. Cool the reactants to 20°C. Quench the reaction mixture with a solution of NH4OH (1.5 V) and H2O (3 V). Separate the organic phase and wash twice with a solution of NH4OH (1.5 V) and H2O (3 V). Wash the organic phase sequentially with aqueous EDTA solution (5% w / w, 3 V) and aqueous NaCl solution (5% w / w, 4 V). Concentrate the organic phase to approximately 1–2 V under reduced pressure at 40–45°C. Dilute the concentrated mixture with 2–MeTHF (6 V) and H2O (3 V). Adjust the pH of the system to pH = 1.1–1.3 with aqueous HCl solution (2 N). Wash the aqueous layer twice with 2–MeTHF (4 V) (this allows for the recycling of compound G contained in the 2–MeTHF solution; detailed procedure is described below). Adjust the pH of the aqueous phase to 8–9 by slowly adding aqueous NaOH solution (30% w / w). Concentrate the aqueous phase under reduced pressure until no fraction remains, obtaining crude compound H. The crude compound H was dissolved in MeCN (2 V) and stirred at 40–50 °C for 3 h. H2O (1 V) was added to the solution at 35–45 °C, followed by the addition of seed crystals of compound H (0.5% w / w). The mixture was stirred at 35–45 °C for 2 h, and H2O (2 V) was added to it. The mixture was cooled to 15–25 °C. The slurry was filtered and the filter cake was washed with MeCN / H2O (2 V, v / v = 2:3). The wet filter cake was dried in a forced-air oven (N2, 45–50 °C) for 12 h to obtain compound H as a brown solid (51.9% yield relative to compound G, 98.8% a / a).

[0332] To recover compound G, a 2-MeTHF solution containing compound G was added to the reactor and washed twice with NaOH aqueous solution (0.1 N, 1 V). The organic phase was separated and H2O (1 V) was added. Adjust the pH of the aqueous phase to 1-1 using an aqueous HCl solution (3N).2. Separate the organic phase and concentrate it under vacuum at 45°C until dry. Dissolve the residue in IPA (1 V) and concentrate to 1–2 V. Dilute the concentrated residue with IPA (8 V). Add IPA (2 V) containing D-camphor-10-sulfonic acid (CSA, 1.2 equiv) dropwise to the reaction mixture at 15–20°C. Stir the reaction mixture at 15–20°C for 2 h and then at 50–55°C for 2 h. Add compound G CSA seed crystals (0.1% w / w) to the reaction mixture at 43 ± 3°C and then stir at 43 ± 3°C for 2 h. Further cool the solution to -6 ± 3°C and stir at that temperature for 4 h. Filter the resulting suspension and wash with IPA (1.0 V). Resuspend the wet filter cake in IPA (3 V) and stir at 40°C for 60 min. Cool the mixture to 5°C and stir at 5°C for 60 min. The slurry was filtered and washed with IPA (1 V) to obtain a wet filter cake in the form of compound G CSA salt. The resulting wet filter cake (1.0 equiv) was suspended in MTBE (3.5 V) at 20 ± 5 °C, followed by the addition of H2O (4 V). The pH was adjusted to pH 8–10 by slowly adding an aqueous solution of NaOH (30% w / w). The mixture was stirred at 20 ± 5 °C for 0.5 h. The organic phase was separated and washed with H2O (3 x 3 V). The organic layer was concentrated to 1–2 V. The concentrated mixture was diluted with n-heptane (3 V) and further concentrated under reduced pressure to approximately 2–3 V. Solvent exchange with n-heptane was repeated twice. The concentrated mixture was diluted with n-heptane (3 V) and stirred at 55 ± 5 °C for 3 h. The mixture was cooled to 20 ± 5 °C and stirred at 20 ± 5 °C for 1 h. The slurry was filtered and washed with n-heptane (1.2 V). The wet filter cake was dried in a Cone dryer at 45-80°C to obtain compound G, which was a grayish-white solid.

[0333] Condition 2: DCM (15 V), TEA (4.0 equiv), 1-cyclopropylpiperazine (2.5 equiv), and Cu(OAc)2 (1.25 equiv) were added to the reactor at 15-20°C with stirring, using TEA as the base. 21% O2 was bubbled under the surface at 20-25°C for 1 h. At 10–20 °C, DCM containing (S)-3-(5-bromo-2-(2-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol (1.0)Add equiv) to the reaction mixture. Heat the reaction mixture at 25–35 °C, during which 21% O2 is bubbled under the surface and stirred under these conditions for at least 5 h. Take a sample for HPLC analysis. Cool the reaction mixture to 20 °C. Quench the reaction mixture with an aqueous solution of NH4OH (1.5 V) and H2O (3 V). Separate the organic phase and wash sequentially with N-acetylcysteine ​​(1 N, 3 V) and H2O (2 V). Concentrate the organic phase to approximately 1–2 V under reduced pressure at 40–45 °C. Dilute the concentrated mixture with 2-MeTHF (6 V) and H2O (3 V). Adjust the pH of the system to pH = 1.1–1.3 with an aqueous solution of HCl (2 N). Wash the aqueous layer twice with 2-MeTHF (4 V). Adjust the pH of the aqueous phase to 8–9 by slowly adding an aqueous solution of NaOH (2 N). Concentrate the aqueous phase under reduced pressure until no fraction remains, to obtain crude compound H. The crude compound H was dissolved in MeCN (2 V) and stirred at 40–50 °C for 3 h. H₂O (3 V) was slowly added to the solution at 35–45 °C. The mixture was cooled to 15–25 °C and stirred further for 3 h. The slurry was filtered, and the filter cake was washed with MeCN / H₂O (2 V, v / v = 2:3). The wet filter cake was dried under vacuum to obtain compound H as a brown solid.

[0334] LCMS (ESI+): C30H39BrF3N4O2(M+H) Calculated value: 623.2; Experimental value: 622.9 1H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 2.0 Hz, 1 H), 7.95 (s, 1 H), 7.62 (d, J = 4.4 Hz, 1 H), 7.36 (d, J = 2.0 Hz, 1 H), 7.18 (s, 1 H), 5.32–5.26 (m, 1 H), 4.61–4.47 (m, 2 H), 3.85–3.83 (m, 1 H), 3 0.19 (s, 4H), 3.03–3.02 (m, 2H), 2.91 (s, 3H), 2.68 (s, 4H), 2.60 (s, 1H), 2.30–2.27 (m, 1H), 1.66 (s, 1H), 1.32 (d, J = 2.0 Hz, 3H), 0.62 (s, 6H), 0.45–0.35 (m, 4H).

[0335] Alternative synthesis of a mixture of transisomers of compound G—(S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol moiety 1-synthesis (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionic acid 2,2,2-trifluoroethyl ester was added to the reactor at 10-20°C with DMF (770 L, 7 V) and (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1H-indol-3-yl)-2,2-dimethylpropionic acid (110 kg, 255.0 mol, 1.0 equiv). The reaction mixture was cooled to 0–5 °C, and Cs₂CO₃ (208.0 kg, 637.0 mol, 2.5 equiv) was added, followed by dropwise addition of 2,2,2-trifluoroethyl trifluoromethanesulfonate (148.0 kg, 637.0 mol, 2.5 equiv). The reaction mixture was stirred at 0–5 °C for 16 h. After completion, the reaction was quenched by adding H₂O at 0–20 °C. The compound was extracted with MTBE (550 L, 5 V). The organic layer was separated, and the aqueous layer was extracted with MTBE (550 L, 5 V). The combined organic layers were washed with brine (330 L, 3 V). The organic layer was concentrated to 190–380 L under reduced pressure at 40–50 °C. The concentrated residue was diluted with THF (220 L, 2 V) and further concentrated to approximately 1–2 V under reduced pressure. The previous unit operation was repeated until the MTBE content in the THF solution was less than 1% w / w. The resulting THF solution of (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionic acid 2,2,2-trifluoroethyl was used directly in the next step (123.6 kg analytical weight, 97.5% yield).

[0336] LCMS (ESI+): C25H26BrF6N2O3(M+H) Calculated value: 597.1; Experimental value: 597 1H NMR (400 MHz, DMSO-d6) δ 8.76 (d, J = 4.8 Hz, 1H), 7.83-7.77 (m, 2H), 7.68-7.64 (m, 1H), 7.57-7.52 (m, 1H), 7.43-7.39 (m, 1H), 4.50-4.43 (m,2H), 3.59 (t,J= 12 Hz, 1H), 3.04 (s, 1H), 2.92 (s, 2H), 2.88 (s, 2H), 2.73 (s, 2H), 1.76-7.73 (m, 1H), 1.37 (d,J= 8.0 Hz, 1H), 1.23 (d,J= 8.0 Hz, 1H), 0.98 (d,J= 20 Hz, 6H).

[0337] A mixture of 2-synthetic compounds G-transisomers - (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylprop-1-ol was added to a reactor at 15-20°C to a THF solution (123.6 kg analytical weight, 207.0 mol, 1.0 equiv) of 2,2,2-trifluoroethyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionic acid 2,2,2-trifluoroethyl ester. The reaction mixture was heated to 50–60 °C and LiBH4 (2 M in THF, 155.7 L, 311.0 mol, 1.5 equiv) was added. The reaction mixture was stirred at 60–65 °C for 24 h. Samples were taken for HPLC analysis. After completion, the reaction was quenched with aqueous HCl (0.5 M, 618.0 L, 5 V) at 10–20 °C. HCl aqueous solution (0.5 M) was added to adjust the pH to 4–5. MTBE (618 L, 5 V) was added to the mixture and stirred at 10–20 °C for 0.5 h. The organic layer was separated and the aqueous layer was washed with MTBE (370 L, 3 V). The combined organic layers were washed sequentially with aqueous NaOH (247 L, 2 V) and brine (247 L, 2 V). The organic layer was concentrated under reduced pressure at 40-45°C to obtain a mixture of the transisomers of compound G (93.5 kg, 90.1% a / a).

[0338] Substitutional synthesis of the mixture of transisomers of compound G: (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionate methyl ester EtOH solution (1.0 equiv, 5 V) was added to the reactor at 15-20°C under N2. The mixture was then charged with N2 at 5°C.CaCl2 (1.25 equiv) was added in batches, followed by NaBH4 (1.5 equiv) at 5°C. The reaction mixture was stirred at 20–30°C for 14 h. The reaction mixture was sampled for HPLC analysis. The reaction was quenched by adding HCl aqueous solution (3 M) dropwise until pH 1–2 was reached. NaOH aqueous solution (30% w / w) was added to the mixture to adjust the pH to between 4 and 5. The resulting mixture was concentrated and precipitated from water. The resulting filter cake was washed with water and dried under vacuum at 45°C.

[0339] DMF (7 V), compound K (1.0 equiv) and K3PO4 (1.4 equiv) were added to the reactor. The reactor was flushed with DMF (454.6 kg, 7 V). The reaction mixture was cooled to 15–20°C and 2,2,2-trifluoroethyl trifluoromethanesulfonate (2 equiv) was added. The reaction mixture was stirred at 15–20°C for 16 h. The reaction mixture was sampled for HPLC analysis. Afterward, the reaction was quenched at 15–20 °C with AcOH (64.4 kg, 2.0 equiv) and H₂O (1435.0 kg, 6 V). The resulting mixture was extracted twice with MTBE (5 V, 2 V). The combined organic layers were concentrated to 3–4 V at 35–45 °C. The concentrated mixture was diluted with IPA (4 V) and further concentrated under reduced pressure at 35–45 °C to approximately 3–4 V. The resulting EtOH solution of methyl (S)-3-(5-bromo-2-(2-(1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropionate (compound G) in a 1.4:1 diastereomer mixture was used directly in the next step.

[0340] Example 15 - Synthesis of Compound 1 - (2S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)-N-((22S,63S,4S)-12-(5-(4-cyclopropylpiperyl)) Specification 111 / 123 pages 123 CN 122270438 A (1-azinyl)-2-((S)-1-methoxyethyl)pyridinyl-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)pyridazine-cycloundecane-4-yl)acetamide moiety 1- Synthesis of (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-3-((3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropoxy)carbonyl)tetrahydropyridazin-1(2H)-yl)-3-oxopropyl)morpholine-4-carboxylic acid tert-butyl ester. DCM (746.15 kg, 9 V) was charged into the reactor, followed by the addition of compound H (67.35 kg, 108.0 mol, 1.0 equiv), compound J (64.57 kg, 124.0 mol, 1.15 equiv), and 1-hydroxybenzotriazole (14... A solution of DCM (1 V) containing 0.02 kg (103.8 mol, 1.0 equiv), DMAP (6.50 kg, 53.2 mol, 0.5 equiv), DIPEA (27.00 kg, 208.9 mol, 2.0 equiv), and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (29.65 kg, 154.7 mol, 1.5 equiv) was prepared. The reaction mixture was stirred at 15 ± 5 °C for 16 h. The reaction was monitored by HPLC and the reaction was confirmed to be complete. The reaction mixture was quenched with an aqueous solution of NaHCO3 (7% w / w, 7.3% w / w of compound H) at 15 ± 5 °C. The organic layer was washed with an aqueous solution of 5% w / w citric acid (731.64 kg). The crude organic solution was concentrated to 3.5–4.5 V under reduced pressure at NMT 45 °C. The concentrated residue was further diluted with DCM (511.85 kg, 6 V). The resulting solution was concentrated to 3.5–4.5 V under reduced pressure at NMT 45 °C. The DCM solution of (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-3-((3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropoxy)carbonyl)tetrahydropyridazin-1(2H)-yl)-3-oxopropyl)morpholine-4-carboxylic acid tert-butyl ester was used in the next step without further purification (90.4% Instructions for Use, pages 112 / 123, 124, CN 122270438, A a / a).

[0341] LCMS (ESI+): C55H72BrF3N8O9(M+H) Calculated value: 1125.5; Experimental value: 1125.9 1H NMR (400 MHz, DMSO-d)6, 25℃) δ8.48 (d,J= 2.8 Hz, 1H) , 7.87 (d,J= 2.0 Hz, 1H) , 7.65 (d ,J= 8.7 Hz, 1H) , 7.39 (dd ,J= 8.8, 1.9 Hz, 1H) , 7.33 (m, 5H), 7.22 (d,J= 2.7 Hz, 1H), 7.12 (d,J= 8.8 Hz, 1H), 5.33 (dd,J= 17.0, 8.6 Hz, 1H), 5.07 (m, 2H), 4.98 (s, 2H) , 4.51 (m , 1H) , 3.97 (s , 1H) , 2.88 (s , 3H) , 1.37 (s, 9H) , 1.32 (d ,J= 6.2 Hz, 3H) , 0.78 (s, 3H) , 0.73 (s, 3H) , 0.42 (m, 2 H) , 0.34 (m, 2H).

[0342] Part 2-Synthesis of (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester Condition 1: Post-treatment with K2CO3 The reactor is charged with (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-3-((3-(5-bromo-2-yl) A solution of tert-butyl morpholine-4-carboxylate (64.1 kg, 1.0 eq) in DCM was prepared and cooled to 5 ± 5 °C under nitrogen. TFA (237.10 kg, 2.4 V) was added dropwise to the reactor while maintaining the internal temperature at 5 ± 5 °C. The reaction mixture was stirred at 10 ± 5 °C for 1 h and then at 15 ± 5 °C for at least 4 h. The reaction was monitored by HPLC, indicating completion. A K₂CO₃ aqueous solution (731.65 kg, 40% w / w) was added to the reaction mixture to adjust the pH to the range of 7–10, while maintaining the temperature at 10 ± 10 °C. H₂O (57.50 kg, 4 V) was added to the mixture and stirred for 0.5 seconds.h. Separate the organic layer and wash with H2O (452.5 kg, 7 V). Exchange the solvent in the solution with MTBE (470.9 kg, 10 V) by distillation. Add the resulting MTBE solution of the crude product to n-heptane (872.05 kg, 20 V) in another reactor at 15 ± 5 °C for 6 h. Stir the mixture at 15 ± 5 °C for at least 3 h. Filter the slurry and wash the wet filter cake with n-heptane (87 kg, 2 V). The crude solid was dried under vacuum at 25±5℃ for 10 h to obtain (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester (103.72 kg, 94.1% a / a, 90.1% yield, after 2 steps).

[0343] Condition 2: Post-treatment with NaOH: (S)-2-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-3-((3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-3-yl)-2,2-dimethylpropoxy)carbonyl)tetrahydropyridazin-1(2H)-yl)-3-oxopropyl)morpholine-4-carboxylic acid tert-butyl ester DCM solution (1.0 eq) was added to the reactor and cooled to 5±5°C under nitrogen. (Instructions 113 / 123 pages 125 CN 122270438 A) TFA (237.10 kg, 2.4 V) was added dropwise to the reactor while maintaining the internal temperature at 5 ± 5 °C. The reaction mixture was stirred at 10 ± 5 °C for 1 h, followed by stirring at 15 ± 5 °C for at least 4 h. The reaction was monitored by HPLC to indicate completion. NaOH aqueous solution (20% w / w, 5X starting material) was added to the reaction mixture and stirred at 5 °C for 1 h. 5% NaOH aqueous solution (4X starting material) was added to adjust the pH to the range of 8–10 while maintaining the temperature at 0–10 °C. H2O (4 V) was added to the mixture and stirred for 0.5 h. The organic layer was separated and washed with H2O (7 V). The solvent in the solution was exchanged with MTBE (470.9 kg, 10 V) by evaporation. The resulting MTBE solution of the crude product was added to n-heptane (20 V) in another reactor at 15 ± 5 °C for 6 h. The mixture was stirred at 15 ± 5 °C for at least 3 h. Filter the slurry and use n-heptane (2V) Wash the wet filter cake. Dry the crude solid under vacuum at 25±5℃ for 10 h to obtain (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholin-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester, which is a white solid.

[0344] LCMS (ESI+): C50H64BrF3N8O7(M+H) Calculated value: 1025.4; Experimental value: 1025.8 1H NMR (400 MHz, DMSO-d 6, 25℃) δ 8.48 (d, J = 2.9 Hz, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.65 (d, J = 8.7 Hz, 1H), 7.39 (dd, J = 8.8, 1.9 Hz, 1H), 7.35 (m, 5H), 7.21 (d, J = 2.8 Hz, 1H), 7.03 (d, J = 8.8 Hz, 1H), 5 .33 (dd,J= 17.1, 8.6 Hz, 1H), 5.05 (m, 2H), 4.98 (s, 2H), 4.49 (dq,J= 15.7, 9.3 Hz, 1H), 3.98 (s, 1H), 2.88 (s, 3H) , 1.32 (d ,J= 6.2 Hz, 3H) , 0.78 (s, 3H) , 0.73 (s, 3H) , 0.43 (m, 2H) , 0.34 (m, 2H).

[0345] Partial 3-synthesis of ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-11H-4-yl)carbamate. Condition 1: P(t-Bu)3•HBF4 ligands were added to the reactor along with toluene (591.90 kg, 25 V) and (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)- 3-((S)-morpholino-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester (27.00 kg, 1.0 eq), anhydrous K3PO4 (27.90 kg, 5.0 eq). The mixture was degassed (O2 content ≤ 1000 ppm) by bubbling with nitrogen at 20±5 °C for 1 h with stirring. Pd2(dba)3•CHCl3 (1.41 kg, 0.047 eq), P(t-Bu)3•HBF4 (1.98 kg, 0.105 eq), and H2O (4.35 kg, 9.0 equiv) were charged into the reactor and the reaction mixture was further degassed by bubbling with nitrogen for 1 h. The reaction mixture was stirred at 90±5 °C for 11 h. The reaction was monitored by HPLC (pages 114 / 123, CN 122270438 A), indicating completion. The crude reaction mixture was cooled to 20±10℃ and filtered. EtOAc (449.90 kg, 10 V) was added to the filtrate. The solvent in the resulting solution was concentrated to 3-5 V under reduced pressure at NMT 55℃. Solvent exchange with EtOAc was repeated twice. The concentrated solution was washed sequentially with H2O (99.50 kg, 2 V), NaCl aqueous solution (10% w / w, 26.95 kg, 0.5 V), and HCl aqueous solution (1% w / w, 17.50 kg, 0.4 V) at 5±10℃. The organic phase was separated and extracted twice with HCl aqueous solution (1% w / w, 4 V and 3 V). EtOAc (183.25 kg, 4 V) and n-heptane (35.05 kg, 1 V) were added to the combined aqueous phase at 5 ± 10 °C. NaCl aqueous solution (25% w / w, 33.45 kg, 0.6 V) was added dropwise to the mixture over 0.5 h. After stirring for 30 min, the aqueous phase was separated. The aqueous phase was concentrated under vacuum at NMT 35 °C to completely remove any organic solvent residues. The resulting suspension was stirred at 5 ± 10 °C for at least 2 h. The slurry was filtered and the wet filter cake was washed with NaCl aqueous solution (10% w / w, 10.35 kg, 0.2 V). The filter cake was collected as the HCl salt of the product. EtOAc (468.10 kg, 10 V) and H₂O (262.00 kg, 5 V) were added to the reactor. The filter cake was added to the reactor with stirring at 5 ± 10 °C. A 40% w / w, 20.95 kg aqueous solution of K₂CO₃ was added to the solution to adjust the pH to 8–10. The organic phase was separated, and the aqueous phase was washed with EtOAc (234.00 kg, 5 V). The combined phases were washed with a 10% w / w, 278.00 kg, 5 V aqueous solution of NaCl.The organic phase was concentrated to 5.5–6.5 V under vacuum at NMT 45°C. The concentrated organic phase was extracted twice with an aqueous HCl solution (1% w / w, 103.70 kg, 2 V) and the aqueous phase was collected. The aqueous phase was concentrated at NMT 35°C under reduced pressure (approximately 0.09 MPa) and a solid precipitate was formed. The residue was stirred at 5 ± 10°C for 1 h, and then an aqueous NaCl solution (25% w / w, 15.50 kg, 0.25 V) was added. The resulting mixture was stirred at 5 ± 10°C for 2 h. The slurry was filtered and the filter cake was washed with an aqueous NaCl solution (10% w / w, 16.10 kg, 0.3 V). The filter cake was collected as the HCl salt of the product.

[0346] EtOAc (234.00 kg, 5 V) and H2O (130.50 kg, 2.5 V) were added to the reactor. The filter cake was added to the reactor under agitation at 5±10℃. A K₂CO₃ aqueous solution (40% w / w) was added to the solution to adjust the pH to 8–10. The organic phase was separated, and the aqueous phase was washed with EtOAc (140.60 kg, 3 V). The combined organic phases were washed with NaCl aqueous solution (10% w / w, 154.85 kg). The organic phase was concentrated to 1.5–2.5 V under vacuum at NMT 45℃. The concentrated residue of the product was charged into a reactor containing n-heptane (704.55 kg, 21 V) at 20±5℃ for 2 h. The slurry was agitated at 20±5℃ for 2 h. The slurry was filtered, and the filter cake was washed with n-heptane (34.85 kg, 2 V). The wet filter cake was dried in an oven at 30±5℃ under vacuum for 10 h to obtain a white solid ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61, 62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-4-yl)carbamate (98.6% a / a, 20.60 kg, 44.6% yield).

[0347] LCMS (ESI+): C50H63F3N8O7(M+H) Calculated value: 945.5; Experimental value: 945.9 1H NMR (300 MHz, DMSO-d 6, 25℃) 8.46 (dd, J = 2.8 Hz, 1H), 7.57 (d, J = 9.0 Hz, 1H), 7.35 (m, 5H), 7.17 (m, 2H), 7.08 (d,J= 9.1 Hz , 1H) , 6.98 (s , 1H) , 5.28 (m, 2 H) , 4.97 (m, 2H) , 4.71 (m, 1H) , 3.33 (s, 3H) , 1.33 (d ,J= 6.1 Hz, 3H) , 0.83 (s, 3H), 0.43 (m, 7H), 0.34 (s, 2H).

[0348] Condition 2: Qphos ligand specification 115 / 123 pages 127 CN 122270438 A Toluene (496.90 kg, 35 V), (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholino-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester (15.01 kg, 14.6 mol, 1.0 eq), anhydrous K3PO4 (16.21 kg, 76.4 mol, 5.0 eq). The mixture was degassed (O2 content ≤ 1000 ppm) by bubbling with nitrogen under stirring at 20±5 °C for 1 h. Pd2(dba)3•CHCl3 (745.60 g, 0.7 mol, 0.047 eq), 1,2,3,4,5-pentaphenyl-1'-(di-tert-butylphosphino)ferrocene (1.09 kg, 1.5 mol, 0.105 eq), and H2O (1.65 kg, 91.7 mol, 6.3 equiv) were charged into the reactor and the reaction mixture was further degassed (O2 content ≤ 1000 ppm) by bubbling with nitrogen for 1 h. The reaction mixture was stirred at 90±5 °C for 10 h. The reaction was monitored by HPLC, indicating completion. The crude reaction mixture was cooled to 20±10 °C and filtered. The filtrate was concentrated to 3–5 V. Add EtOAc (358.00 kg, 10 V) to the concentrated residue. Concentrate the resulting solution to 3–5 V under reduced pressure at NMT 55 °C. Repeat solvent exchange with EtOAc twice. Wash the concentrated solution sequentially with H₂O (95.00 kg, 2.5 V) and aqueous HCl (1% w / w, 25.00 kg, 0.5 V) at 0 ± 5 °C. Separate the organic phase and extract twice with aqueous HCl (1% w / w, 4 V and 3 V). Add 25% NaCl aqueous solution (27.10 kg, 0.80 V) and EtOAc (137.15 kg, 10 V) to the combined aqueous phase at 5 ± 10 °C.The organic phase was separated by stirring for 30 min. The aqueous phase was concentrated under vacuum at NMT 35 °C to completely remove any organic solvent residue. The resulting solution was stirred at 5 ± 10 °C for at least 2 h. A sample of the mother liquor was taken to obtain the product concentration. The slurry was filtered and the wet filter cake was washed with an aqueous NaCl solution (10% w / w, 10.00 kg, 0.3 V). The filter cake was collected as the HCl salt of the product. EtOAc (340.45 kg, 10 V) and H2O (185.50 kg, 5 V) were added to the reactor. The filter cake was added to the reactor with stirring at 5 ± 10 °C. An aqueous K2CO3 solution (40% w / w, 16.00 kg) was added to the solution to adjust the pH to the 8–10 range. The organic phase was separated and the aqueous phase was washed with EtOAc (174.00 kg, 5 V). The combined organic phases were washed with an aqueous NaCl solution (10% w / w, 190.50 kg, 5 V). The organic phases were concentrated to 2–3 V under vacuum at NMT 45 °C. The concentrated residue of the product was charged into a reactor containing n-heptane (539.60 kg, 21 V) at 20 ± 5 °C for 2 h. The slurry was stirred at 20 ± 5 °C for 2 h. The slurry was filtered and the filter cake was washed with n-heptane (2 V). The wet filter cake was dried in an oven at 30 ± 5 °C under reduced pressure for 10 h to obtain the crude product. The crude product (13.38 kg, 14.2 mol, 1.0 equiv) was charged into a reactor containing H₂O (91.50 kg, 6.8% w / w) and HCl (2.59 kg, 0.19% w / w) at 5 ± 5 °C. The mixture was stirred at 5 ± 5 °C for 2 h and then added dropwise with a 25% NaCl aqueous solution (23.90 kg, 1 V). The resulting slurry was filtered and the wet filter cake was washed with a 10% NaCl aqueous solution (31.40 kg, 2 V). The wet filter cake was suspended in H₂O (94.00 kg, 7 V) and EtOAc (96.45 kg, 8 V) at 5 ± 10 °C. The pH of the aqueous layer was adjusted to 8–10 with a K₂CO₃ aqueous solution (40% w / w, 10.65 kg). The organic layer was separated and washed with EtOAc (36.40 kg, 3 V). The organic layers were combined and washed with a 10% w / w NaCl aqueous solution (67.10 kg, 5% w / w). The organic phase was concentrated to 2–3 V under vacuum at NMT 45 °C. The concentrated residue was diluted with EtOAc (60.75 kg, 5 V) and the resulting mixture was concentrated to 2–3 V under vacuum at NMT 45°C. The concentrated residue was then concentrated at 20 ± 5°C. (Instructions 116 / 123, page 128, CN 122270438)The residue was added dropwise to n-heptane (208.10 kg, 21 V) and the mixture was stirred for 2 h. The slurry was filtered and the wet filter cake was washed with n-heptane (18.65 kg, 2 V). The wet filter cake was dried in an oven under vacuum at 30±5℃ for 6 h to obtain a white solid ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-cycloundecane-4-yl)carbamate (11.91 kg, 87.6% yield).

[0349] Condition 3: Using dioxane as a solvent, 1,4-dioxane (58.0 L, 35 V), (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholino-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester (1.50 kg, 1.5 mol, 1.0 eq) and anhydrous K3PO4 (1.03 kg, 4.8 mol, 3.3 eq) were charged into the reactor. The mixture was degassed (O2 content ≤ 1000 ppm) by bubbling with nitrogen under stirring at 20±5 °C for 1 h. P(t-Bu)3PdG3 (93 g, 0.15 mol, 0.11 eq), P(t-Bu)3·HBF4 (46.2 g, 0.15 mol, 0.11 eq), and H2O (205 mL, 6.6 equiv) were charged into the reactor and the reaction mixture was further degassed (O2 content ≤ 1000 ppm) by bubbling with nitrogen for 1 h. The reaction mixture was stirred at 90±5 °C for 10 h. The reaction was monitored by HPLC, indicating completion. The crude reaction mixture was cooled to 20±10 °C and filtered. The filter cake was washed with 1,4-dioxane (1.6 L, 1 V). The filtrate was concentrated to give a crude product as a yellow solid. The solid was dissolved in 2-MeTHF (13.8 L, 6 V) and aqueous HCl (0.08 M, 5 V). The organic layer was separated and washed with aqueous HCl (0.08 M, 4 V). The organic layer was extracted twice with aqueous HCl (0.3 M, 5 V) at -15 °C. The solid was then extracted twice with 2-MeTHF (10.5 L, 5 V) at -15 °C.V) Wash the combined aqueous layers twice. Add solid Na₂HCO₃ to the aqueous layers at 10–20 °C to adjust the pH to 7. Extract the resulting aqueous solution twice with 2-MeTHF (10.5 L, 10 V). Wash the combined organic layers sequentially with aqueous acetic acid (0.04 M, 10.5 L, 10 V) and 25% w / w NaCl aqueous solution (10.5 L, 10 V). The organic layer obtained by concentration under reduced pressure yielded a yellow solid ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-4-yl)carbamate (1.056 kg, 91.6% a / a).

[0350] Condition 4: Pd(t-Bu3P)2 catalyst and new separation process specification 117 / 123 pages 129 CN 122270438 A Anisole (20 V) and (S)-1-((S)-2-(((benzyloxy)carbonyl)amino)-3-((S)-morpholino-2-yl)propionyl)hexahydropyridazine-3-carboxylic acid 3-(5-bromo-2-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-3-yl)-2,2-dimethylpropyl ester (1.0 eq) were charged into the reactor. The mixture was degassed (O2 content ≤ 1000 ppm) by bubbling nitrogen under agitation at 20±5 °C for 1 h. Cs₂CO₃ (3 equiv) and bis(tri-tert-butylphosphine)palladium (0.1 equiv) were charged into the reactor and the reaction mixture was further degassed by bubbling with nitrogen for 1 h (O₂ content ≤ 1000 ppm). The reaction mixture was stirred at 90 ± 5 °C for 11 h. The reaction was monitored by HPLC and the reaction was confirmed to be complete. The crude reaction mixture was cooled to 20 ± 10 °C and the mixture was filtered. The filter cake was washed with anisole (3.5% w / w) and EtOAc (3.6% w / w). The filtrate was concentrated to 3–5 V under reduced pressure at NMT 55 °C. Solvent exchange with EtOAc was repeated twice. The EtOAc solution was extracted twice with aqueous HCl solution (1% w / w, 4 V and 3 V) to form the HCl salt of the product. Alternatively, the EtOAc solution was extracted with p-toluenesulfonic acid or trichloroacetic acid to form the corresponding toluenesulfonate or trichloroacetate of the product. The combined aqueous phases were inoculated with the HCl salt of the product (0.01% w / w) at 5°C. The mixture was then...The mixture was stirred at 5°C for 5 h, followed by the addition of 25% NaCl aqueous solution (0.6% w / w). The resulting mixture was stirred at 5°C for another 5 h. The slurry was filtered, and the wet filter cake was washed with 10% NaCl aqueous solution (0.2% w / w). The wet filter cake was suspended in EtOAc (9% w / w) and H2O (6.5% w / w). 40% K2CO3 solution was added at 0–10°C to adjust the pH to 8–10. After stirring for 1 h, the organic phase was separated, and the aqueous phase was extracted with EtOAc (4.5% w / w). The organic layers were combined and purified via a silica gel pad (100–200 mesh, 0.7% w / w). The precipitate from the silica gel was concentrated to 2.5 V under reduced pressure and NMT at 40°C. The concentrated solution was added dropwise to n-heptane (1.43% w / w) at 20°C. The resulting mixture was stirred at 20°C for 5 h. The slurry was filtered and the filter cake was washed with n-heptane (1.4% w / w). The wet filter cake was dried in an oven at 30°C under reduced pressure for 8 h to obtain a white solid ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2 (4,2)-morpholina-1(5,3)-indolia-6(1,3)pyridazine-11H-4-yl)carbamate.

[0351] Part 4-Synthesis (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-5,7-dione Specification 118 / 123 pages 130 CN 122270438 A Condition 1: THF (211.60 kg, 12) is charged into the pressure reactor using K2CO3 base. V), ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-4-yl) benzyl carbamate (19.89 kg, 21 mol, 1.0 equiv), 5%The reaction mixture was prepared by stirring at 0.2 mPa H2 and 20–25 °C with Pd / C (20% w / w, 4.25 kg) and K2CO3 (2.97 kg, 21 mol, 1 equiv). The atmosphere was purged several times and refilled with fresh H2 gas, as monitored by HPLC. After the reaction was complete, the atmosphere was purged with N2 gas and the reaction mixture was filtered through a diatomaceous earth mat. The filtrate was then concentrated to 3–5 V under reduced pressure at NMT 40 °C and diluted with EtOAc (124.65 kg, 7 V). The mixture was further concentrated to 3–5 V under reduced pressure at NMT 40 °C. The concentrated residue was diluted with EtOAc (127.40 kg, 7 V) and washed with a 5% w / w N-acetyl-L-cysteine ​​aqueous solution (198.05 kg, 10% w / w). The mixture was filtered through a fluid filter to remove the Pd complex. Add 5% w / w Na₂CO₃ aqueous solution to the filtrate to adjust the pH of the aqueous layer to 7.4–8.0. Separate the organic layer and wash sequentially with 5% w / w Na₂CO₃ aqueous solution and 5% w / w NaCl aqueous solution. Concentrate the organic layer to 3–5 V under reduced pressure at NMT 40 °C. Dilute the concentrated residue with EtOAc (5 V) and further concentrate the resulting solution to 3–5 V under reduced pressure at NMT 40 °C. Repeat the solvent exchange process twice. After 2 h, add MTBE (58.65 kg, 4 V) to the solution of the crude product in EtOAc, followed by adding product seed crystals (0.01% w / w, 200 g) at 20 ± 10 °C. Stir the mixture at 20 ± 10 °C for 2 h. Dilute the mixture with MTBE (5 V) and concentrate to 4–6 V under reduced pressure at NMT 40 °C. Solvent exchange with MTBE was repeated three times. The concentrated solution was cooled to 0±5℃ and stirred for 2 h. The slurry was filtered and the wet filter cake was washed with MTBE (20.85 kg, 1 V). The filter cake was dried under vacuum at 30±5℃ in an oven to obtain a white solid (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-5,7-dione (13.79 kg, 99.7% a / a, 84% yield).

[0352] LCMS (ESI+):Calculated value of C42H57F3N8O5(M+H): 811.4; Experimental value: 811.5. 1H NMR (400 MHz, DMSO-d6, 25℃) δ 8.45 (d, J = 2.8 Hz, 1H), 7.58 (d, J = 8.9 Hz, 1H), 7.15 (d, J = 2.7 Hz, 1H), 7.09 (d, J = 8.9 Hz, 1H), 6.99 (s, 1H), 5.35 (dd, J = 16.5, 8.5 Hz, 1H), 5.06 (d, J = 12.2 Hz, 1H), 4.77 (dd, J = 16.6, 9.1 Hz, 1H). Hz, 1H), 3.33 (s, 3H), 1.33 (d,J= 6.1 Hz, 3H), 0.79 (s, 3H), 0.42 (m, 4H), 0.34 (s, 3H).

[0353] Condition 2: K2CO3-free base and includes citric acid washing. Instructions 119 / 123 pages 131 CN 122270438 A. THF (12 V), ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-11H-4-yl)carbamate (1.0 equiv) and 5% w / w Pd / C (40%) were added to the pressure reactor. (w / w, 4.25 kg). The mixture was stirred at 0.2 mPa H2 and 20–25 °C. During the reaction, the atmosphere was exchanged several times by venting and refilling with fresh H2 gas. After the reaction was complete, as monitored by HPLC, the atmosphere was exchanged with N2 gas and the reaction mixture was filtered through a diatomaceous earth mat. MTBE (14 V) and 0.2% citric acid aqueous solution (6 V) were added to the filtrate. The organic phase was separated and washed with 3% w / w Na2CO3 aqueous solution (6 V). The organic layer was separated and washed with H2O (3 V). The organic phase was separated and MTBE (10 V) was added. The resulting mixture was concentrated to 3–4 V. Solvent exchange with MTBE was repeated twice. The concentrated residue was diluted with MTBE (1 V). The slurry was stirred for 3 h and then filtered. The filter cake was washed with MTBE (1 V) and dried in an oven at 30–35 °C.The wet filter cake was dried under reduced pressure at ℃ to obtain a white solid (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-cycloundecane-5,7-dione.

[0354] Condition 3: Using 2-MeTHF as solvent, 2-MeTHF (12 V), ((22S,63S,4S)-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-4-yl) benzyl carbamate (1.0 equiv), 5% w / w Pd / C (20% w / w) and K2CO3 (1 equiv) were added to the autoclave without solvent exchange. The mixture was stirred at 20–25 °C in the presence of 0.2 mPa H2. During the reaction, the atmosphere was exchanged several times by venting and refilling with fresh H2 gas. As monitored by HPLC, after the reaction was complete, the atmosphere was exchanged with N2 gas and the reaction mixture was filtered through a diatomaceous earth mat (pages 120 / 123, CN 122270438 A), and the wet filter cake was washed with 2-MeTHF (5–7 times). The filtrate was washed with 5% w / w N-acetyl-L-cysteine ​​aqueous solution (10 V) at 10 °C. The mixture was filtered. The filtrate was adjusted to pH 7.4–8.0 with 5% w / w Na2CO3 aqueous solution. The organic phase was separated. The aqueous phase was extracted with 2-MeTHF (3 V). The organic phases were combined and washed sequentially with 5% w / w Na2CO3 aqueous solution (5 V) and 5% w / w NaCl aqueous solution (5 V). The organic phase was concentrated to 7 V at NMT 35 °C and exchanged twice with 2-MeTHF (10 V). Product seed crystals (0.01 X) and n-heptane (14 V) were added dropwise to the concentrated solution of the crude product at 25 °C. The resulting slurry was stirred at 5 °C for 9 h. The slurry was filtered and the wet filter cake was washed with 2-MeTHF / n-heptane (v / v = 1:2, 3 V). The wet filter cake was dried in an oven at 30 °C under vacuum for 39 h. A white solid (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-5,7-dione (611.53 g, 97.1% analytical grade, 85.5% yield).

[0355] Partial 5-synthetic compound 1-(2S)-2-cyclopentyl-2-((S)-7-((2R,3R)-3-cyclopropyl-1-methylaziridinium-2-carbonyl)-2,7-diazaspiro[4.4]non-2-yl)-N-((22S,63S,4S)-12-(5-(4-cyclopropylpiperazine- 1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-hexacycloundecane-4-yl)acetamide. Condition 1: HOPO / EDCl coupling, 2-MeTHF / IPA / heptane crystallization. EtOAc (94.80 kg, 10 V) and (22S,63S,4S)-4-amino-12-(5-(4-) Cyclopropylpiperazine-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-cycloundecane-5,7-dione (10.50 kg, 1.0 equiv), compound (5.30 kg, 1.1 equiv), DIPEA (6.60 kg, 4.0 equiv), DMAP (0.80 kg, 0.5 equiv), HOPO (0.23 kg, 0.15 equiv) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.50 kg, 1.40 equiv). The mixture was stirred at 20–30 °C for 9 h until the reaction was complete. The crude reaction mixture was added to a reactor containing H₂O (84.00 kg, 8 V) at 10 ± 5 °C. The two-phase mixture was stirred at 10 ± 5 °C for 1 h, and the organic phase was separated. The mixture was then reacted sequentially with an aqueous solution of NaHCO₃ (5 wt%, 105.20 kg) and an aqueous solution of NaCl (2% w / w, 105.20 kg).The organic phase was washed with H2O (5 V) at 5 ± 5 °C and the aqueous phase was adjusted to pH 5.5–6.1 with 5% w / w citric acid aqueous solution (32.65 kg). The organic phase was separated. H2O (10.05 kg, 1 V) was added to the organic phase at 5 ± 5 °C and 1% w / w Na2CO3 aqueous solution (7.70 kg) was added to adjust the pH to 9–10. The organic phase was separated and washed with 2% w / w NaCl aqueous solution (84.00 kg). The organic phase was concentrated to 3–5 V under vacuum at NMT 40 °C. The concentrated residue was diluted with 2-MeTHF (10 V) and the mixture was concentrated to 3–5 V under vacuum at NMT 40 °C. Solvent exchange with 2-MeTHF was repeated three times. A 2-MeTHF solution of the crude product was obtained and further purified by recrystallization. IPA (9.70 kg, 1.0 V) and n-heptane (20.85 kg, 2.5 V) were added to a 2-MeTHF solution of compound 1 at 5 ± 5 °C, followed by the addition of compound 1 seed crystals (5% w / w, 0.92 kg). The slurry was stirred at 5 ± 5 °C for 12 h. The slurry was filtered and the wet filter cake was dried in an oven under vacuum at 35 ± 10 °C to give compound 1 (10.84 kg, 71.6% yield). Instruction manual, pages 121 / 123, 133, CN 122270438 A

[0356] LCMS (ESI+): C63H88F3N11O7(M+H) Calculated value: 1168.7; Experimental value: 1169.2 1H NMR (400 MHz, CD3OD, 25℃) δ 8.42 (d, J = 2.4 Hz, 1H), 7.47 (d, J = 9.2 Hz, 1H), 7.28 (d, J = 1.2 Hz, 1H), 7.12 (m, 1H), 7.10 (m, 1H), 5.67 (d, J = 9.2 Hz, 1H), 5.00 (m, 1H), 4.71 (m, 1H) , 4.44 (d ,J= 12.8 Hz, 1H) , 4.06 (d ,J= 2.0 Hz, 1H) , 3.91‑3.28 (m , 15H) , 3.19 (s , 3H) , 2.89‑2.66 (m , 14 H) , 2.34 (d ,J= 14.0 Hz, 3H) , 2.24‑1.41 (m, 25 H) , 0.88 (s, 3H) , 0.64 (m, 1 H) , 0.54 (s, 3H) , 0.50‑0.26 (m, 8 H).

[0357] Condition 2: PyBOP / Oxyma coupling, 4-dioxane / IPE crystallization: DMF (60 mL, 6 V), compound E (5.10 g, 1.05 equiv), DIPEA (7.95 g, 5.0 equiv), (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65, 66-Hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indola-6(1,3)-pyridazine-cycloundecane-5,7-dione (10 g, 1.0 equiv). The mixture was cooled to 0 ± 5 °C and oxyma (1.58 g, 0.9 equiv) was added. The reaction mixture was cooled to -10 ± 5 °C and DMF (20 mL, 2 V) containing PyBOP (7.69 g, 1.2 equiv) was added. The mixture was stirred at -10 ± 5 °C for 1 h until the reaction was complete. H2O (35 mL, 3.5 V) was added to the reaction mixture and the mixture was filtered through a microporous filter. Another portion of H2O (54 V) was added to the filtrate at 20 ± 5 °C and the mixture was stirred for 0.5 h. The slurry was filtered and the filter cake was washed with H2O (5 V). The wet filter cake was dried under a stream of N2 to give a pink solid. The pink solid was dissolved in IPOAc (7 V) and MTBE (5 V). The solution was washed twice sequentially with H2O (12 V), 0.83% w / w NaH2PO4 aqueous solution (12 V), and 3% w / w NaHCO3 aqueous solution (12 V). The organic layer was concentrated to dryness. The residue was dissolved in IPOAc (15 V) and slurried with 9% charcoal at rt for 1 h. The mixture was filtered. The filtrate was concentrated to dryness to give crude compound 1. The crude product was dissolved in 1,4-dioxane (2.5 V) and IPE (1.5 V) at 25 °C. The solution was cooled to 10 °C and stirred for 2 h. IPE (1 V) was added dropwise to the solution over 8 h, followed by the addition of another portion of IPE (5 V) over 10 h. The resulting slurry was filtered and the filter cake was washed with 1,4-dioxane / IPE (v / v = 2.5:7.5, 2 V). A second recrystallization of the solid was performed by dissolving it in 1,4-dioxane (2.5 V) and IPE (1.5 V) at 25 °C. IPE (1 V) was added dropwise to the solution over 0.5 h at 25 °C. The solution was then cooled to 10 °C. Compound 1 was inoculated, and the mixture was stirred at 10 °C for 3 h. Another portion of IPE was added after 12 h.(3.5 V) was added to the mixture and the slurry was stirred for 6 h. The slurry was filtered and the filter cake was washed with 1,4-dioxane / IPE (v / v=2.5:5.5, 2 V). The wet filter cake was dried under vacuum at 30 °C to give compound 1 as a light yellow solid.

[0358] Condition 3: HOBt / EDCl Coupling Instructions, pages 122 / 123, 134, CN 122270438 A. EtOAc (58.11 kg, 10 V), (22S,63S,4S)-4-amino-12-(5-(4-cyclopropylpiperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-10,10-dimethyl-11-(2,2,2-trifluoroethyl)-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-morpholina-1(5,3)-indolia-6(1,3)-pyridazine-cycloundecane-5,7-dione (6.40 kg, 1.0 equiv), and compound (3.50 kg, 1.1 equiv) were sequentially added to the reactor. The mixture was prepared with DIPEA (4.20 kg, 4.0 equiv), DMAP (481.9 g, 0.5 equiv), HOBt (74.9 g, 0.07 equiv), and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.15 kg, 1.40 equiv). The mixture was stirred at 20–30 °C for 9 h until the reaction was complete. The crude reaction mixture was added to a reactor containing H₂O (52.60 kg, 8 V) at 10 ± 5 °C. The two-phase mixture was stirred at 10 ± 5 °C for 1 h, and the organic phase was separated. The organic layer was washed sequentially with aqueous NaHCO₃ solution (5 wt%, 64.00 kg, 6.4% w / w) and aqueous NaCl solution (2% w / w, 64.28 kg, 6.4% w / w). H₂O (5 V) was added to the organic phase at 5 ± 5 °C, and the aqueous phase was adjusted to pH 5.0–5.6 with 5% w / w citric acid aqueous solution (1.35 kg). The organic phase was separated. H₂O (6.55 kg, 1 V) was added to the organic phase at 5 ± 5 °C, and 1% w / w NaCO₃ aqueous solution (10.30 kg) was added to adjust the pH to 9–10. The organic phase was separated and washed with 2% w / w NaCl aqueous solution (51.20 kg, 8% w / w). The organic phase was concentrated to 3–5 V under vacuum at NMT 40 °C. The concentrated residue was diluted with 2-MeTHF (10 V), and the mixture was concentrated to 3–5 V under vacuum at NMT 40 °C. Solvent exchange with 2-MeTHF was repeated three times.The crude product in 2-MeTHF solution was further purified by recrystallization. Heptane (15.00 kg, 2.5 V) was added to the 2-MeTHF solution of compound 1 at 5 ± 5 °C, followed by the addition of compound 1 seed crystals (5% w / w, 0.92 kg). An antisolvent consisting of 2-MeTHF (7.90 kg, 1.33 V), IPA (3.70 kg, 0.67 V), and heptane (122.25 kg, 26 V) was added to the compound 1 solution. The slurry was stirred at 5 ± 5 °C for 12 h. The slurry was filtered and the wet filter cake was dried in an oven under vacuum at 35 ± 10 °C to give compound 1 as a white solid (6.24 kg, 98.4% a / a, 67.7% yield).

[0359] Other Embodiments Although the invention has been described in conjunction with its specific embodiments, it should be understood that further modifications are possible and this application is intended to cover any variations, uses, or adaptations of the invention that generally follow the principles of the invention and are included within the scope of known or conventional practice in the field to which the invention pertains, and that may be applied to the essential features set forth herein.

[0360] All publications, patents, and patent applications are incorporated herein by reference in their entirety as if each individual publication, patent, or patent application were specifically and individually instructed to be incorporated by reference. Specification 123 / 123 Page 135 CN 122270438 A 1. A method for separating enantiomers of a compound of formula I, the method comprising the steps of: a) contacting the compound of formula I with a chiral acid to form a diastereomeric salt of the compound of formula I; and b) separating each diastereomeric salt of each diastereomeric salt, thereby separating the enantiomers of the compound of formula I wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 2. The method of claim 1, wherein the chiral acid comprises a chiral carboxylic acid. 3. The method of claim 1 or 2, wherein the separation step (b) comprises recrystallization. 4. The method of any one of claims 1 to 3, wherein the compound of formula I is further described by formula Ia: . 5. A barium salt of a compound of compound A: . 6. The barium salt of claim 5, wherein the barium salt is a 2:1 (compound A:barium) salt. 7. A method for preparing the barium salt of claim 5 or 6, the method comprising the steps of: a) condensing a compound of formula IIa with compound C to form a compound of formula IIb; b) cyclizing the compound of formula IIb with a compound of formula IIc to form a compound of formula IId; c)Hydrolyzing the compound of formula IId to form the compound of formula IIe: (Claims amended according to Article 19 of the Treaty, page 1 / 14, 136, CN 122270438 A) d) Methylating the compound of formula IIe to form the compound of formula IIf: ; and e) Hydrolyzing the compound of formula IIf to form a barium salt of the compound A: wherein R2 and R3 are each independently an optionally substituted C1-C6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-C10 aryl, and LG is a detached group selected from the group consisting of halogen, trifluoromethanesulfonate, methanesulfonate, and toluenesulfonate. 8. The method of claim 7, wherein the methylation step (d) comprises contacting the compound of formula IIe with MeB(OH)2. 9. The method of claim 7 or 8, wherein the hydrolysis step (e) comprises contacting the compound of formula IIf with barium hydroxide. 10. A method for preparing a compound of formula III or a salt thereof: Formula III The method comprises the steps of: a) coupling a compound of formula IIId with a compound of formula I to form a compound of formula IIIe; and b) deprotecting the compound of formula IIIe to form the compound of formula III: Claims amended under Article 19 of the Treaty 2 / 14 page 137 CN 122270438 A Wherein n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen protecting group; Each R4 is independently an optionally substituted C1-C6 alkyl or optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 11. The method of claim 10, wherein the compound of formula IIId is prepared by a method comprising the steps of: A1) combining the compound of formula IIIa with the compound of formula IIIb to form the compound of formula IIIc; B1) reducing the compound of formula IIIc to form the compound of formula IIid; wherein Q is MgBr, MgCl, MgI, or Li. 12. The method of claim 11, wherein the reduction step (B1) comprises contacting the compound of formula IIIc with a ketone reductase. 13. The method of claim 10, wherein the compound of formula IIId is prepared by the steps of: A2) converting the compound of formula IIIf to form the compound of formula IIIg; and B2) reacting the compound of formula IIIg (e.g., esterification) to form the compound of formula IIId. 14. The method of claim 13, wherein the conversion step (A2) comprises forming a diazonium salt of the compound of formula IIIf.15. A method for preparing a compound of formula IV or a salt thereof: Claims amended according to Article 19 of the Treaty, page 3 / 14, 138 CN 122270438 A. The method comprises the steps of: a) coupling a compound of formula III with a barium salt of compound A to form a compound of formula IVa; and b) hydrolyzing said compound IVa to form said compound IV: wherein n and m are each independently 0, 1, 2, 3, 4 or 5; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 16. The method of claim 15, wherein the coupling step (a) comprises contacting the compound of formula III and the barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH). 17. A method for preparing a compound of formula VIII or a salt thereof: The method comprises the steps of: a) esterifying and N-alkylating a compound of formula VIIIa to form a compound of formula VIIIb; b) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers; and c) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 18. A method for preparing a compound of formula VIII or a salt thereof: The method comprises the steps of: a) esterifying a compound of formula VIIIa to form a compound of formula VIIId; b) N-alkylating the compound of formula VIIId to form the compound of formula VIIIb; c) reducing the compound of formula VIIIb to form a compound of formula VIIIc in the form of a mixture of transisomers; and d) separating the mixture of transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 19. A method for preparing a compound of formula IX or a salt thereof: The method comprises the steps of: a) borated a compound of formula VIII to form a compound of formula X; and b) coupling the compound of formula X with the compound of formula XI to form a compound of formula XII; wherein R12 is a halogen;R13 is an optionally substituted C1-C6 alkyl group; and R14 is an optionally substituted 3- to 6-membered cycloalkyl group. 20. A method for preparing a compound of formula XIV or a salt thereof: The method comprises the steps of: a) reacting a compound of formula XV with a compound of formula XII to form a compound of formula XVI: (Claims amended according to Article 19 of the Treaty, page 6 / 14, 141, CN 122270438 A); b) removing PGa from the compound of formula XVI to form a compound of formula XVII: ; c) cyclizing the compound of formula XVII with a transition metal catalyst to form a compound of formula XVIII: ; d) removing PGa from the compound of formula XVIII to form a compound of formula XIX: ; and e) coupling the compound of formula XIX with the compound of formula IV to form a compound of formula XIV: ; wherein n and m are each independently 0, 1, 2, 3, 4 or 5; (Claims amended according to Article 19 of the Treaty, page 7 / 14, 142, CN 122270438 A) X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-; R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; PGa is an acid-instability protecting group; and PGb is an acid-stability protecting group. 21. The method of claim 20, wherein step (a) comprises reacting the compound of formula XII with the compound of formula XV in the presence of a carbodiimide coupling agent, an anti-racemic agent, and a base. 22. The method of claim 20 or 21, wherein step (c) further comprises contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII. 23. The method of any one of claims 20 to 22, wherein step (e) comprises coupling the compound of formula XIX with the compound of formula IV in the presence of a coupling agent, an anti-racemic agent, and a base. 24. A compound of formula IIf-1: or a salt thereof, wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl; R20 is hydrogen or a C1-6 alkyl optionally substituted with one or two substituents independently selected from CN, OH, C1-6 alkoxy, and C1-6 haloalkyl; R21 is OH, halogen, CN, C1-6 alkyl, C1-6 alkoxy, or C1-6 haloalkyl; and q is 0, 1, or 2. 25. A method for preparing a compound of formula IIf: the method comprising alkylating a compound of formula IIe with a methylating agent: wherein R2 is an optionally substituted C1-6 alkyl, optionally substituted C3-6 cycloalkyl, or optionally substituted C6-10 aryl. 26. The method of claim 25, wherein the methylating agent comprises trimethylcycloboroxane. Claims amended according to Article 19 of the TreatyPage 143 of 8 / 14 CN 122270438 A 27. A method for preparing a compound of formula IVa: The method comprises coupling a compound of formula III to a compound of formula IIf, wherein n and m are each independently 0, 1, 2, 3, 4 or 5; each R4 is independently an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; each R5 is independently an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 28. The method of claim 27, wherein the coupling comprises contacting the compound of formula III and the compound of formula IIf with a base. 29. A method for preparing a compound of formula VIIIc: The method comprises N-alkylating a compound of formula VIIIe with a compound having a structure having R13-LG1: wherein each R12 is a halogen; each R13 is an optionally substituted C1-C6 alkyl group; and LG1 is a detachable group. 30. A method for preparing a compound of formula VIIIe: Claims amended according to Article 19 of the Treaty, page 9 / 14, 144 CN 122270438 A The method comprises reducing a compound of formula VIIIf with a reducing agent: wherein R12 is a halogen; and R13 is an optionally substituted C1-C6 alkyl group. 31. The method of claim 30, wherein the reducing agent is NaBH4. 32. A compound having the structure of formula XVIII: wherein R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl group, optionally substituted C1-C3 alkoxy group, optionally substituted 3- to 6-membered cycloalkyl group, optionally substituted 3- to 6-membered heterocyclic alkyl group; p is 0, 1, 2 or 3; and PGb is an acid-stabilizing protecting group. 33. The compound of claim 32, wherein the compound has the following structure: Claims amended according to Article 19 of the Treaty, page 10 / 14, 145 CN 122270438 A. 34. A compound having the structure of formula XVII: wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; each R15 is independently hydrogen, a halogen, an optionally substituted C1-C3 alkyl group, an optionally substituted C1-C3 alkoxy group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 6-membered heterocyclic alkyl group; p is 0, 1, 2, or 3; and PGb is an acid-stabilizing protecting group. 35. The compound of claim 34, wherein the compound has the following structure: [Claims amended according to Article 19 of the Treaty]Page 11 / 14, 146, CN 122270438, A 36. A compound having the structure of formula XVI: wherein R12 is a halogen; R13 is an optionally substituted C1-C6 alkyl group; R14 is an optionally substituted 3- to 6-membered cycloalkyl group; PGa is an acid-instability protecting group; each R15 is independently hydrogen, halogen, optionally substituted C1-C3 alkyl group, optionally substituted C1-C3 alkoxy group, optionally substituted 3- to 6-membered cycloalkyl group, optionally substituted 3- to 6-membered heterocyclic alkyl group; p is 0, 1, 2, or 3; and PGb is an acid-stabilizing protecting group. 37. The compound of claim 36, wherein the compound has the following structure: . 38. A compound having the structure of compound H: . 39. A compound having the structure of formula IV: (Claims amended according to Article 19 of the Treaty, page 12 / 14, 147 CN 122270438 A) wherein n and m are each independently 0, 1, 2, 3, 4, or 5; R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-. 40. The compound of claim 39, wherein the compound of formula IV is compound E: . 41. A compound having the structure of formula IVa: wherein n and m are each independently 0, 1, 2, 3, 4, or 5; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; R5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl, or an optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2, and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-. 42. The compound of claim 41, wherein the compound has the following structure: 43. A compound having the structure of Formula III: (Claims amended according to Article 19 of the Treaty, page 13 / 14, 148 CN 122270438 A) wherein n and m are each 1; R4 is an optionally substituted C1-C6 alkyl or an optionally substituted C6-C10 aryl; and R5 is an optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl or optionally substituted 3- to 6-membered heterocycloalkyl; and X1, X2 and X3 are each independently -CH2-, -CHF-, -CF2-, -C(O)- or -O-. 44. The compound of claim 43, wherein the compound of Formula III is compound D: .45. A compound having the structure of formula IId: wherein R2 and R3 are each independently a optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, or optionally substituted C6-C10 aryl. 46. The compound of claim 45, wherein the compound has the following structure: [Claims amended according to Article 19 of the Treaty, page 14 / 14, 149, CN 122270438 A]

Claims

1. A method for separating the enantiomers of compound I, Formula I The method includes the following steps: a) Contact the compound of formula I with a chiral acid to form a diastereomeric salt of the compound of formula I; and b) Separate each diastereomer of the said diastereomeric salt, thereby separating the enantiomers of compound I. in n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen-protecting group; and X 1 X 2 and X 3 Each can be independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

2. The method of claim 1, wherein the chiral acid comprises a chiral carboxylic acid.

3. The method of claim 1 or 2, wherein the separation step (b) comprises recrystallization.

4. The method according to any one of claims 1 to 3, wherein the compound of formula I is further described by formula Ia: 。 5. A barium salt of compound A: 。 6. The barium salt of claim 5, wherein the barium salt is a 2:1 (compound A: barium) salt.

7. A method for preparing the barium salt as described in claim 5 or 6, the method comprising the following steps: a) Condensate compound IIa with compound C to form compound IIb: b) Cyclate the compound of formula IIb with the compound of formula IIc to form the compound of formula IId: c) Hydrolyze the compound of formula IId to form the compound of formula IIe: d) Methylating the compound of formula IIe to form a compound of formula IIf: ;and e) Hydrolyze the compound of formula IIf to form a barium salt of compound A: in R 2 and R 3 Each is independently a optionally substituted C1-C6 alkyl group, or an optionally substituted C 3-6 cycloalkyl or optionally substituted C6-C 10 Aryl, and LG is a detached group selected from the group consisting of: halogen, trifluoromethanesulfonate, methanesulfonate, and toluenesulfonate.

8. The method of claim 7, wherein the methylation step (d) comprises contacting the compound of formula IIe with MeB(OH)2.

9. The method of claim 7 or 8, wherein the hydrolysis step (e) comprises contacting the compound of formula IIf with barium hydroxide.

10. A method for preparing a compound of formula III or a salt thereof: The method includes the following steps: a) Couple a compound of formula IIId with a compound of formula I to form a compound of formula IIIe: ;and b) Deprotecting the compound of formula IIIe to form the compound of formula III: in n and m are each independently 0, 1, 2, 3, 4 or 5; PG is a nitrogen-protecting group; Each R 4 Independently, it is an optionally substituted C1-C6 alkyl group or an optionally substituted C6-C alkyl group. 10 Aryl; R 5 It is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl, or an optionally substituted 3- to 6-membered heterocycloalkyl; and X 1 X 2 and X 3 Each can be independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

11. The method of claim 10, wherein the IIId compound is prepared by a method comprising the following steps: A1) Combining compound IIIa with compound IIIb to form compound IIIc: B1) Reduction of compound IIIc to form compound IIid: . Where Q is MgBr, MgCl, MgI or Li.

12. The method of claim 11, wherein the reduction step (B1) comprises contacting the compound of formula IIIc with a ketone reductase.

13. The method of claim 10, wherein the compound of formula IIId is prepared by the following steps: A2) Transform the compound of formula IIIf to form the compound of formula IIIg: ;and B2) React the compound of formula IIIg (e.g., esterify) to form the compound of formula IIId: 。 14. The method of claim 13, wherein the conversion step (A2) comprises forming a diazonium salt of the compound of formula IIIf.

15. A method for preparing a compound of formula IV or a salt thereof: The method includes the following steps: a) Couple compound III with the barium salt of compound A to form compound IVa: ;and b) Hydrolyze the compound of formula IVa to form the compound of formula IV: in n and m are each independently 0, 1, 2, 3, 4 or 5; R 4 Optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 Aryl; R 5 It is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl, or an optionally substituted 3- to 6-membered heterocycloalkyl; and X 1 X 2 and X 3 Each can be independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

16. The method of claim 15, wherein the coupling step (a) comprises contacting the compound of formula III and the barium salt of compound A with chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (TCFH).

17. A method for preparing a compound of formula VIII or a salt thereof: The method includes the following steps: a) Esterification and N-alkylation of compound VIIIa to form compound VIIIb: ； b) Reduce the compound of formula VIIIb to form compound of formula VIIIc in the form of a mixture of transisomers: ;and c) Separate a mixture of the transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; Where R 12 It is a halogen; and R 13 It is an optional substituted C1-C6 alkyl group.

18. A method for preparing a compound of formula VIII or a salt thereof: The method includes the following steps: a) Esterifying the compound of formula VIIIa to form the compound of formula VIIId: ； b) N-alkylating the compound of formula VIIId to form the compound of formula VIIIb: ； c) Reduce the compound of formula VIIIb to form compound of formula VIIIc in the form of a mixture of transisomers: ;and d) Separate a mixture of the transisomers of formula VIIIc to obtain a stereochemically pure compound of formula VIII; Where R 12 It is a halogen; and R 13 It is an optional substituted C1-C6 alkyl group.

19. A method for preparing a compound of formula IX or a salt thereof: The method includes the following steps: a) Boronize the compound of formula VIII to form the compound of formula X: ;and b) Couple the compound of formula X to the compound of formula XI to form the compound of formula XII: ； Where R 12 It is a halogen; R 13 It is an optionally substituted C1-C6 alkyl group; and R 14 It is an optional substituted 3- to 6-membered cycloalkyl group.

20. A method for preparing a compound of formula XIV or a salt thereof: The method includes the following steps: a) Reacting compound XV with compound XII to form compound XVI: ； b) Removal of PG from compound XVI a Compounds of formula XVII: ； c) Cycling of compound XVII using a transition metal catalyst to form compound XVIII: ； d) Removal of PG from the compound of formula XVIII b Compounds of formula XIX are formed as follows: ;and e) Couple the compound of formula XIX to the compound of formula IV to form the compound of formula XIV: ； Where n and m are each independently 0, 1, 2, 3, 4 or 5; X 1 X 2 and X 3 Each can be independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-; R 12 It is a halogen; R 13 The substituted C1-C6 alkyl group is optional; R 14 The substituted 3- to 6-membered cycloalkyl groups are optional. PG a It is an acid-instability protecting group; and PG b It serves as a protective group for acid stability.

21. The method of claim 20, wherein step (a) comprises reacting the compound of formula XII with the compound of formula XV in the presence of a carbodiimide coupling agent, an anti-racemic agent and a base.

22. The method of claim 20 or 21, wherein step (c) further comprises contacting the compound of formula XVIII with hydrochloric acid to form a hydrochloride salt of the compound of formula XVIII.

23. The method of any one of claims 20 to 22, wherein step (e) comprises coupling the compound of formula XIX to the compound of formula IV in the presence of a coupling agent, an anti-racemic agent and a base.

24. A compound of formula IIf-1: or its salt, wherein R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-6 cycloalkyl or optionally substituted C 6-10 Aryl; R 20 Hydrogen or optionally hydrogen, or one or two independently selected from CN, OH, C 1-6 Alkoxy and C 1-6 The C-substituent of the haloalkyl group 1-6 alkyl; R 21 For OH, halogen, CN, C 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkyl groups; and q can be 0, 1, or 2.

25. A method for preparing a compound of formula IIf: The method includes alkylating a compound of formula IIe with a methylating agent: Where R 2 C is an optional replacement 1-6 Alkyl, optionally substituted C 3-6 cycloalkyl or optionally substituted C 6-10 Aryl.

26. The method of claim 25, wherein the methylating agent comprises trimethylcycloboroxane.

27. A method for preparing compounds of formula IVa: The method includes coupling a compound of formula III with a compound of formula IIf. Where n and m are each independently 0, 1, 2, 3, 4 or 5; Each R 4 Independently, it is an optionally substituted C1-C6 alkyl group or an optionally substituted C6-C alkyl group. 10 Aryl; Each R 5 Independently, it is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3- to 6-membered cycloalkyl, or an optionally substituted 3- to 6-membered heterocycloalkyl; and X 1 X 2 and X 3 Each can be independently -CH2-, -CHF-, -CF2-, -C(O)-, or -O-.

28. The method of claim 27, wherein the coupling comprises contacting the compound of formula III and the compound of formula IIf with a base.

29. A method for preparing compounds of formula VIIIc: The method includes using R 13 -LG 1 The structure of the compound is used to N-alkylate the compound of formula VIIIe: Among them, each R 12 It is a halogen; Each R 13 It is an optionally substituted C1-C6 alkyl group; and LG 1 For detachment from the base.

30. A method for preparing compounds of formula VIIIe: The method includes reducing compound VIIIf with a reducing agent: Where R 12 It is a halogen; and R 13 It is an optional substituted C1-C6 alkyl group.

31. The method of claim 30, wherein the reducing agent is NaBH4.