TW202412792A

TW202412792A - p38α-MK2 INHIBITOR, AND PHARMACEUTICAL COMPOSITION AND USE THEREOF

Info

Publication number: TW202412792A
Application number: TW112132651A
Authority: TW
Inventors: 楊方龍; 黃悦; 傅東林; 俞立挺; 范景榮; 王思勤; 金磊
Original assignee: 大陸商長春金賽藥業有限責任公司
Priority date: 2022-08-30
Filing date: 2023-08-29
Publication date: 2024-04-01

Abstract

The present disclosure disclosed a compound as shown in formula I, and racemate, stereoisomer, tautomer, isotope marker, solvate, polymorphism, pharmaceutically acceptable salt or prodrug thereof. The compound has a good p38α-MK2 inhibitory effect, which can be used for treatment or prevention diseases related to p38α-MK2 and for prepare drugs of such diseases or conditions.

Description

p38α-MK2 inhibitor compounds, pharmaceutical compositions and uses thereof

本發明要求享有下述專利申請的優先權：於2022年08月30日向中國國家知識產權局提交的，專利申請號為202211058712.X，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2022年10月27日向中國國家知識產權局提交的，專利申請號為202211328642.5，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2022年11月25日向中國國家知識產權局提交的，專利申請號為202211494113.2，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2022年12月09日向中國國家知識產權局提交的，專利申請號為202211589267.X，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2023年01月06日向中國國家知識產權局提交的，專利申請號為202310020787.7，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2023年02月01日向中國國家知識產權局提交的，專利申請號為202310050723.1，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2023年02月21日向中國國家知識產權局提交的，專利申請號為202310145391.5，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案；於2023年08月04日向中國國家知識產權局提交的，專利申請號為202310979990.7，名稱為「p38α-MK2抑制劑化合物、藥物組合物及其應用」的先申請案。所述先申請案的全文通過引用的方式結合於本發明中。 This invention claims the priority of the following patent applications: The prior application filed with the National Intellectual Property Administration of China on August 30, 2022, with patent application number 202211058712.X, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on October 27, 2022, with patent application number 202211328642.5, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on November 25, 2022, with patent application number 202211494113.2, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on December 9, 2022, with patent application number 20221589267.X, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on January 6, 2023, with patent application number 202310020787.7, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on February 1, 2023, with patent application number 202310050723.1, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on February 21, 2023, with patent application number 202310145391.5, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications"; The prior application filed with the National Intellectual Property Administration of China on August 4, 2023, with patent application number 202310979990.7, entitled "p38α-MK2 inhibitor compounds, drug compositions and their applications". The full text of the prior application is incorporated into the present invention by reference.

本發明屬於藥物化合物領域，具體涉及p38α-MK2抑制劑化合物、藥物組合物及其用途。The present invention belongs to the field of pharmaceutical compounds, and specifically relates to p38α-MK2 inhibitor compounds, pharmaceutical compositions and uses thereof.

p38激酶，是絲裂原活化蛋白激酶(mitogen-activated protein kinase，MAPK)家族的成員之一，參與著許多的細胞生理和病理的過程，包括細胞的凋亡、細胞應激、細胞週期和機體的炎症反應。哺乳動物細胞中有四種不同的p38異構物，分別為p38α、p38β、p38γ和p38δ。P38α生理相關性最大，它和p38β廣泛分佈於所有細胞。p38δ主要位於睾丸、胰腺、小腸和CD4+T細胞。p38γ主要在肌肉中表達。p38 MAPK啟動後，由其介導的下游磷酸化調節主要為兩大類蛋白，一類為轉錄因子，如p53、ATF2、Elk1、MEF2和C/EBPβ，另一類為蛋白激酶，包括MK2(也稱為MAPKAP2)、MSK1、MNK1和MNK2。p38 kinase, a member of the mitogen-activated protein kinase (MAPK) family, is involved in many cellular physiological and pathological processes, including cell apoptosis, cell stress, cell cycle and the body's inflammatory response. There are four different p38 isoforms in mammalian cells, namely p38α, p38β, p38γ and p38δ. P38α has the greatest physiological relevance, and it and p38β are widely distributed in all cells. p38δ is mainly located in the testis, pancreas, small intestine and CD4+T cells. p38γ is mainly expressed in muscle. After p38 MAPK is activated, the downstream phosphorylation regulators mediated by it are mainly two major types of proteins, one is transcription factors, such as p53, ATF2, Elk1, MEF2 and C/EBPβ, and the other is protein kinases, including MK2 (also known as MAPKAP2), MSK1, MNK1 and MNK2.

儘管有幾種p38α和p38β的抑制劑已經進入了臨床研究，然而這些抑制劑由於下游通路過於複雜，存在毒副作用大、有效性差的問題，所以大部分的臨床開發都已經中止。2004年，Davidson等人(Biochemistry, 2004, 43, 11658)通過穩態的動力學和熱力學分析，發現p38α-MK2複合物非常穩定，其解離常數Kd = 6 nM，並發展了一個小分子CMPD1，能夠選擇性的抑制p38α-MK2訊號通路(Ki ^app= 330 nM)，而對p38α-ATF2的影響很小(Ki ^app＞ 20 μM)，這為開發選擇性p38α-MK2抑制劑奠定了基礎。鑒於p38α-MK2訊號通路在調節炎症因子(IL-1β、IL-6、TNFα)和負責炎症的酶(COX-2、iNOS、MMP)中起重要作用，2019年Confluence等報導了一種特異性的p38α抑制劑CDD-450，其能夠選擇性阻斷p38α對促炎激酶MK2的活化，並通過促進IL-1β、TNFα、IL-6 mRNA的降解來緩解大鼠的關節炎。該分子目前進入臨床2期，用於治療風濕性關節炎和化膿性汗腺炎等。 Although several p38α and p38β inhibitors have entered clinical research, most of these inhibitors have been discontinued due to their complex downstream pathways, high toxicity and side effects, and poor efficacy. In 2004, Davidson et al. (Biochemistry, 2004, 43, 11658) found that the p38α-MK2 complex was very stable through steady-state kinetic and thermodynamic analysis, with a dissociation constant of Kd = 6 nM. They also developed a small molecule, CMPD1, which could selectively inhibit the p38α-MK2 signaling pathway (Ki ^app = 330 nM) while having little effect on p38α-ATF2 (Ki ^app ＞ 20 μM), which laid the foundation for the development of selective p38α-MK2 inhibitors. In view of the important role of the p38α-MK2 signaling pathway in regulating inflammatory factors (IL-1β, IL-6, TNFα) and enzymes responsible for inflammation (COX-2, iNOS, MMP), Confluence et al. reported a specific p38α inhibitor CDD-450 in 2019, which can selectively block the activation of p38α on the proinflammatory kinase MK2 and relieve arthritis in rats by promoting the degradation of IL-1β, TNFα, and IL-6 mRNA. The molecule is currently in Phase 2 clinical trials for the treatment of rheumatic arthritis and suppurative hidradenitis.

儘管大量有意義的研究已在該領域進行，目前仍需要繼續研究開發更加有效的小分子p38α-MK2抑制劑。本發明提供了一種新型結構的p38α-MK2抑制劑，並發現具有此類結構的化合物具有良好的活性。Although a lot of meaningful research has been conducted in this field, there is still a need to continue researching and developing more effective small molecule p38α-MK2 inhibitors. The present invention provides a p38α-MK2 inhibitor with a novel structure, and it is found that compounds with this structure have good activity.

為改善上述技術問題，本發明提供了一種式I所示的化合物、其消旋物、立體異構物、互變異構物、同位素標記物、氮氧化物、溶劑化物、多形體、藥學上可接受的鹽或其前藥：式I 其中， X選自CH或N； Y選自化學鍵、C _1-6烷基、氘代C _1-6烷基、4-7員雜環基；環A、環B相同或不同，彼此獨立地選自C _6-14芳基、5-14員雜芳基、3-14員雜環基、C _3-12環烷基；每個R相同或不同，彼此獨立地選自鹵素、CN、羥基、胺基、C _1-6烷基、C _1-6烷氧基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、C _3-8環烷基、-C(R ₁)(R ₂)(R ₃)、-P(=O)(R ₂₁)(R ₂₂)；其中，R ₁、R ₂相同或不同，彼此獨立地選自H、氘、鹵素、CN、羥基、胺基、C _1-6烷基、C _1-6烷氧基、氘代C _1-6烷基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、C _3-8環烷基；或者，R ₁、R ₂與它們連接的原子一起形成任選被1個，2個或更多個R’取代的下列基團：C _3-8環烷基或3-10員雜環基；每個R’相同或不同，彼此獨立地選自H、氘、鹵素、CN、側氧基(=O)、C _1-6烷基、C _1-6烷氧基； R ₃選自H、氘、鹵素、CN、羥基、胺基、C _1-6烷基、C _1-6烷氧基、氘代C _1-6烷基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、HO-C _1-6烷基、C _3-8環烷基、-(CH ₂) _mC(O)R ₈、-(CH ₂) _mC(O)NR ₉R ₁₀、‑S(O) _n‑R ₁₁、-(CH ₂) _mCOOR ₁₂；每個R ₄、R ₇相同或不同，彼此獨立地選自H、氘、鹵素、CN、羥基、胺基、C _1-6烷基、C _1-6烷氧基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、C _3-8環烷基； R ₅、R ₆相同或不同，彼此獨立地選自H、氘、鹵素、CN、C _1-6烷基、C _1-6烷氧基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、C _3-8環烷基； R ₈、R ₉、R ₁₀、R ₁₁、R ₁₂相同或不同，彼此獨立地選自H、氘、無取代或任選被1個，2個或更多個Ra取代的下列基團：C _1-6烷基、C _1-6烷氧基、C _3-8環烷基；每個Ra相同或不同，彼此獨立地選自氘、鹵素、CN、胺基、羥基、側氧基(=O)、C _1-6烷基、C _1-6烷氧基；或者R ₉、R ₁₀與它們連接的N原子一起形成無取代或任選被1個，2個或更多個Rb取代的下列基團：3-10員雜環基或5-10員雜芳基；每個Rb相同或不同，彼此獨立地選自氘、鹵素、CN、側氧基(=O)、羥基、胺基、C _1-6烷基、C _1-6烷氧基； R ₂₁、R ₂₂相同或不同，彼此獨立地選自H、氘、C _1-6烷基、C _1-6烷氧基、鹵代C _1-6烷基、鹵代C _1-6烷氧基、C _3-8環烷基； r選自1、2或3； p、q相同或不同，彼此獨立地選自0、1、2、3或4； m選自0-6的整數； n選自0、1或2。 To improve the above technical problems, the present invention provides a compound represented by Formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof: Formula I wherein, X is selected from CH or N; Y is selected from a chemical bond, a C _1-6 alkyl group, a deuterated C _1-6 alkyl group, a 4-7 membered heterocyclic group; Ring A and Ring B are the same or different and are independently selected from C _6-14 aryl group, a 5-14 membered heteroaryl group, a 3-14 membered heterocyclic group, a C _3-12 cycloalkyl group; each R is the same or different and is independently selected from halogen, CN, hydroxyl, amine, C _1-6 alkyl group, C _1-6 alkoxy group, halogenated C _1-6 alkyl group, halogenated C _1-6 alkoxy group, C _3-8 cycloalkyl group, -C(R ₁ )(R ₂ )(R ₃ ), -P(═O)(R ₂₁ )(R ₂₂ ); wherein, R ₁ , R _{R 2} is the same or different and is independently selected from H, deuterium, halogen, CN, hydroxyl, amine, C _1-6 alkyl, C _1-6 alkoxy, deuterated C _1-6 alkyl, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _3-8 cycloalkyl; or, R ₁ , R ₂ together with the atoms to which they are attached form the following group optionally substituted by 1, 2 or more R': C _3-8 cycloalkyl or 3-10 membered heterocyclic group; each R' is the same or different and is independently selected from H, deuterium, halogen, CN, pendoxy (=O), C _1-6 alkyl, C _1-6 alkoxy; R ₃ is selected from H, deuterium, halogen, CN, hydroxyl, amine, C 1-6 alkyl, C 1-6 alkoxy, deuterated C _1-6 alkyl, halogenated C _1-6 alkoxy, C 3-8 cycloalkyl _wherein the at least one R 4 and R ₇ are _the same or different _and are independently selected from H, deuterium, halogen, _CN _, _hydroxyl _, _amino _, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, _C _3-8 _cycloalkyl ; R ₅ and R ₆ are the same or different _and are independently selected from H, deuterium, halogen, CN, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl _{, halogenated C 1-6} _alkoxy _, C _3-8 cycloalkyl; R ₈ , R ₉ , R ₁₀ , R ₁₁ , and R ₁₂ are the same _or different and are independently selected from H, deuterium, or the following groups which are unsubstituted or optionally substituted with 1 _, ₂ or more Ra: C _1-6 alkyl, C _1-6 alkoxy, or C _3-8 cycloalkyl; each Ra is the same or different and is independently selected from deuterium, halogen, CN, amine, hydroxyl, pendoxy (=O), C _1-6 alkyl, _{or C 1-6} _alkoxy ; or R ₉ , R _{R 10} together with the N atom to which they are attached form the following group which is unsubstituted or optionally substituted by 1, 2 or more R b: a 3-10 membered heterocyclic group or a 5-10 membered heteroaryl group; each R b is the same or different and is independently selected from deuterium, a halogen, CN, a pendoxy group (=O), a hydroxyl group, an amine group, a C _1-6 alkyl group, a C _1-6 alkoxy group; R ₂₁ and R ₂₂ are the same or different and are independently selected from H, deuterium, a C _1-6 alkyl group, a C _1-6 alkoxy group, a halogenated C _1-6 alkyl group, a halogenated C _1-6 alkoxy group, a C _3-8 cycloalkyl group; r is selected from 1, 2 or 3; p and q are the same or different and are independently selected from 0, 1, 2, 3 or 4; m is selected from an integer of 0-6; n is selected from 0, 1 or 2.

根據一些實施方案，X選自N；根據一些實施方案，X選自CH；當X為CH時，R ₇可以取代該位置的H，較佳地，X-R ₇選自C-CN；根據一些實施方案，Y選自C _1-3烷基、氘代C _1-3烷基、4-6員雜環基；根據一些實施方案，Y選自亞甲基、氘代亞甲基(-CD ₂-或-CHD-)、亞乙基(-CH(CH ₃)-或-CH ₂CH ₂-)、-C(CH ₃) ₂-、哌啶基、哌嗪基、四氫吡咯基、氮雜環丁基；根據一些實施方案，Y選自-CH ₂-、-CD ₂-、-CH(CH ₃)-、-C(CH ₃) ₂-或氮雜環丁基；根據一些實施方案，環A、環B相同或不同，彼此獨立地選自C _6-10芳基、5-10員雜芳基；根據一些實施方案，環A、環B相同或不同，彼此獨立地選自苯基或含有1-3個選自N、O、S的雜原子的5-6員雜芳基；根據一些實施方案，環A選自吡啶基或苯基；根據一些實施方案，環B選自嘧啶基、吡嗪基、吡啶基、吡啶氮氧化物基或苯基；根據一些實施方案，每個R相同或不同，彼此獨立地選自鹵素、CN、-C(R ₁)(R ₂)(R ₃)或-P(=O)(R ₂₁)(R ₂₂)；根據一些實施方案，R ₁、R ₂相同或不同，彼此獨立地選自C _1-6烷基、C _3-8環烷基；或者，R ₁、R ₂與它們連接的原子一起形成C _3-8環烷基；根據一些實施方案，R ₁、R ₂彼此獨立地選自甲基、環丙基；或者，R ₁、R ₂與它們連接的原子一起形成環丙基、環丁基、環戊基；根據一些實施方案，R ₃選自CN、羥基、胺基、-C(O)R ₈、-C(O)NR ₉R ₁₀、‑S(O) ₂‑R ₁₁、-COOR ₁₂；其中，R ₈、R ₉、R ₁₀、R ₁₁、R ₁₂相同或不同，彼此獨立地選自H、C _1-6烷基、C _3-8環烷基；或者R ₉、R ₁₀與它們連接的N原子一起形成無取代或任選被1個、2個或更多個Rb取代的下列基團：3-8員雜環基；每個Rb相同或不同，彼此獨立地選自鹵素、CN、側氧基(=O)、羥基、C _1-6烷基、C _1-6烷氧基；根據一些實施方案，R ₈、R ₉、R ₁₀、R ₁₁、R ₁₂相同或不同，彼此獨立地選自H、C _1-3烷基、C _3-6環烷基；或者R ₉、R ₁₀與它們連接的N原子一起形成無取代或任選被1個、2個或更多個Rb取代的下列基團：3-6員雜環基；每個Rb相同或不同，彼此獨立地選自羥基、C _1-6烷基；根據一些實施方案，R ₈、R ₁₁彼此獨立地選自甲基；根據一些實施方案，R ₉、R ₁₀相同或不同，彼此獨立地選自H、甲基、乙基、異丙基、環丙基；或者R ₉、R ₁₀與它們連接的N原子一起形成被1個或2個Rb取代的氮雜環丁基、哌嗪基；每個Rb相同或不同，彼此獨立地選自羥基或甲基；根據一些實施方案，R ₉、R ₁₀相同或不同，彼此獨立地選自H、甲基、乙基、異丙基、環丙基；或者R ₉、R ₁₀與它們連接的N原子一起形成3-羥基-1-氮雜環丁基或4-甲基哌嗪-1-基；根據一些實施方案，R ₃選自CN、羥基、胺基、-CONH ₂、-SO ₂CH ₃、-COCH ₃、-CONHCH ₃、-CONHCH ₂CH ₃、-CONHCH(CH ₃) ₂、-CONH-環丙基、-CON(CH3) ₂、、、-C(CH ₃) ₂(OH)、-COOH、-COOCH ₃、-COOC ₂H ₅。 According to some embodiments, X is selected from N; According to some embodiments, X is selected from CH; when X is CH, _R7 can replace H at that position, preferably, _XR7 is selected from C-CN; According to some embodiments, Y is selected from _C1-3 alkyl, deuterated _C1-3 alkyl, 4-6 membered heterocyclic group; According to some embodiments, Y is selected from methylene _, deuterated methylene ( _-CD2- or -CHD-), ethylene (-CH( _CH3 )- or _-CH2CH2- ), -C( _CH3 ) _2- , piperidinyl, piperazinyl, tetrahydropyrrolyl, azacyclobutyl; According to some embodiments, Y is selected from _-CH2- , _-CD2- , -CH( _CH3 )-, -C( _CH3 ) _2- or azacyclobutyl; According to some embodiments, ring A and ring B are the same or different and are independently selected from C _6-10 aryl, 5-10 membered heteroaryl; According to some embodiments, ring A and ring B are the same or different and are independently selected from phenyl or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S; According to some embodiments, ring A is selected from pyridyl or phenyl; According to some embodiments, ring B is selected from pyrimidyl, pyrazinyl, pyridyl, pyridine nitrogen oxide or phenyl; According to some embodiments, each R is the same or different and is independently selected from halogen, CN, -C(R ₁ )(R ₂ )(R ₃ ) or -P(═O)(R ₂₁ )(R ₂₂ ); According to some embodiments, R ₁ and R ₂ are the same or different and are independently selected from C In some embodiments, R ₁ and R ₂ are independently selected from methyl _and cyclopropyl; or, R ₁ and R ₂ are independently selected from cyclopropyl, cyclobutyl and cyclopentyl; In some embodiments, R ₃ _is selected from CN, _hydroxyl , amine, -C(O)R ₈ , -C(O)NR ₉ R ₁₀ , -S(O) 2 ‑R 11 , -COOR ₁₂ ; wherein R ₈ , R 9 , R ₁₀ , R ₁₁ , and R ₁₂ are the same or different and are _{independently} selected from H, C 1-6 alkyl and C _3-8 cycloalkyl; or, R ₉ , R ₁₀ , R ₁₁ , and R ₁₂ are the same or different and are independently selected from H, C _1-6 alkyl and C _3-8 cycloalkyl; _{R 10} and the N atom to which they are attached form the following groups which are unsubstituted or optionally substituted by 1, 2 or more R b: a 3-8 membered heterocyclic group; each R b is the same or different and is independently selected from halogen, CN, oxo (=O), hydroxyl, C _1-6 alkyl, C _1-6 alkoxy; According to some embodiments, R ₈ , R ₉ , R ₁₀ , R ₁₁ , and R ₁₂ are the same or different and are independently selected from H, C _1-3 alkyl, and C _3-6 cycloalkyl; or R ₉ , R ₁₀ and the N atom to which they are attached form the following groups which are unsubstituted or optionally substituted by 1, 2 or more R b: a 3-6 membered heterocyclic group; each R b is the same or different and is independently selected from hydroxyl, C 1-6 alkyl; According to some embodiments, R ₈ , R 9 , R 10 , R 11 , and R 12 are the same or different and are independently selected from H, C 1-3 alkyl, and C _3-6 cycloalkyl; R ₉ and R ₁₀ are the same or different and are independently selected from H, methyl, ethyl, isopropyl, cyclopropyl; or R ₉ and R ₁₀ together with the N atom to which they are connected form an azacyclobutyl or piperazinyl substituted with ₁ or 2 R b; each R b is the same or different and is independently selected from hydroxyl or methyl; According to some embodiments, R ₉ and R ₁₀ are the same or different and are independently selected from H, methyl, ethyl, isopropyl, cyclopropyl; or R ₉ and R ₁₀ together with the N atom to which they are connected form a 3-hydroxy-1-azacyclobutyl or 4-methylpiperazin-1-yl; According to some embodiments, R ₃ is selected from CN, hydroxyl, amine, -CONH ₂ , -SO ₂ CH ₃ , -COCH ₃ , -CONHCH ₃ , -CONHCH ₂ CH ₃ , -CONHCH(CH ₃ ) ₂ , -CONH-cyclopropyl, -CON(CH3) ₂ , , , -C(CH ₃ ) ₂ (OH), -COOH, -COOCH ₃ , -COOC ₂ H ₅ .

根據一些實施方案，R ₂₁、R ₂₂相同或不同，彼此獨立地選自H、氘、C _1-6烷基、C _1-6烷氧基；根據一些實施方案，R ₂₁、R ₂₂均選自甲基。 According to some embodiments, R ₂₁ and R ₂₂ are the same or different and are independently selected from H, deuterium, C _1-6 alkyl, and C _1-6 alkoxy. According to some embodiments, R ₂₁ and R ₂₂ are both selected from methyl.

根據一些實施方案，每個R ₄、R ₇相同或不同，彼此獨立地選自鹵素、CN、C _1-6烷基；根據一些實施方案，每個R ₄、R ₇相同或不同，彼此獨立地選自鹵素、CN、甲基；根據一些實施方案，R ₄選自甲基或F；根據一些實施方案，R ₇選自F或CN；根據一些實施方案，R ₅、R ₆相同或不同，彼此獨立地選自鹵素、CN、C _1-6烷基；根據一些實施方案，R ₅、R ₆相同或不同，彼此獨立地選自鹵素、甲基；根據一些實施方案，R ₅選自甲基；根據一些實施方案，R ₆選自Cl、Br或甲基；根據一些實施方案，r選自1或2；根據一些實施方案，p選自2或3；根據一些實施方案，q選自1或2；根據一些實施方案，m選自0或1；根據一些實施方案，n選自2。 According to some embodiments, each R ₄ and R ₇ are the same or different and are independently selected from halogen, CN, and C _1-6 alkyl; According to some embodiments, each R ₄ and R ₇ are the same or different and are independently selected from halogen, CN, and methyl; According to some embodiments, R ₄ is selected from methyl or F; According to some embodiments, R ₇ is selected from F or CN; According to some embodiments, R ₅ and R ₆ are the same or different and are independently selected from halogen, CN, and C _1-6 alkyl; According to some embodiments, R ₅ and R ₆ are the same or different and are independently selected from halogen, methyl; According to some embodiments, R ₅ is selected from methyl; According to some embodiments, R ₆ is selected from Cl, Br, or methyl; According to some embodiments, r is selected from 1 or 2; According to some embodiments, p is selected from 2 or 3; According to some embodiments, q is selected from 1 or 2; According to some embodiments, m is selected from 0 or 1; According to some embodiments, n is selected from 2.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式IA 式IA-1 式IA-2 其中，X、Y、環A、環B、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、R ₇、p、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula IA Formula IA-1 Formula IA-2 wherein X, Y, Ring A, Ring B, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式IB 其中，X、Y、R、R ₄、R ₅、R ₆、R ₇、r、p、q具有本文中所述的定義，Z ₁、Z ₂、Z ₃相同或不同，彼此獨立地選自N、N(O)或CH。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula IB wherein X, Y, R, R ₄ , R ₅ , R ₆ , R ₇ , r, p, q have the meanings described herein, and Z ₁ , Z ₂ , and Z ₃ are the same or different and are independently selected from N, N(O) or CH.

根據一些實施方案，Z ₁選自N，Z ₂選自N、N(O)或CH，Z ₃選自N、N(O)或CH； According to some embodiments, _Z1 is selected from N, _Z2 is selected from N, N(O) or CH, and _Z3 is selected from N, N(O) or CH;

根據一些實施方案，Z ₁選自CH，Z ₂選自N、N(O)或CH，Z ₃選自N、N(O)或CH； According to some embodiments, Z ₁ is selected from CH, Z ₂ is selected from N, N(O) or CH, and Z ₃ is selected from N, N(O) or CH;

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式IB-1 其中，X、Y、Z ₁、Z ₂、Z ₃、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、R ₇、p、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula IB-1 wherein X, Y, Z ₁ , Z ₂ , Z ₃ , R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式I-1 式I-2 式I-3 式I-4 式I-5 式I-6 式I-7 式I-8 式I-9 式I-10 。其中，X、Y、環A、環B、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、R ₇、p、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula I-1 Formula I-2 Formula I-3 Formula I-4 Formula I-5 Formula I-6 Formula I-7 Formula I-8 Formula I-9 Formula I-10 . wherein X, Y, Ring A, Ring B, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式IC 其中，X、Y、R、R ₄、R ₅、R ₆、R ₇、r、p、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula IC wherein X, Y, R, R ₄ , R ₅ , R ₆ , R ₇ , r, p, q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式II 式II-1 式II-2 其中，Y、R ₁、R ₂、R ₃、R ₄、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula II Formula II-1 Formula II-2 wherein Y, R ₁ , R ₂ , R ₃ , R ₄ , and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式III 式III-1 其中，R ₁、R ₂、R ₄、R ₉、R ₁₀、Y、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula III Formula III-1 wherein R ₁ , R ₂ , R ₄ , R ₉ , R ₁₀ , Y, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式IV 其中，R ₉、R ₁₀具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula IV wherein R ₉ and R ₁₀ have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式V 式V-1 式V-2 式V-3 其中，Y、R ₁、R ₂、R ₃、R ₄、R ₆、R ₇、Z ₁、Z ₂、Z ₃、R、p、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Type V Type V-1 Type V-2 Formula V-3 wherein Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , Z ₁ , Z ₂ , Z ₃ , R, p, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式VI 式VI-1 式VI-2 式VI-3 其中，Y、R ₄、R ₆、Z ₁、Z ₂、Z ₃、R、q具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula VI Formula VI-1 Formula VI-2 Formula VI-3 wherein Y, R ₄ , R ₆ , Z ₁ , Z ₂ , Z ₃ , R, and q have the meanings described herein.

根據一些實施方案，所述式I所示的化合物選自如下所示的結構：式VII 其中，Y、R ₄、R ₆、R具有本文中所述的定義。 According to some embodiments, the compound shown in Formula I is selected from the structures shown below: Formula VII wherein Y, R ₄ , R ₆ , and R have the definitions described herein.

根據一些實施方案，所述式I所示的化合物選自以下結構：、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、。 According to some embodiments, the compound shown in formula I is selected from the following structures: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

根據一些實施方案，所述式I所示的化合物選自以下結構：、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、。 According to some embodiments, the compound shown in formula I is selected from the following structures: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

本發明還提供一種藥物組合物，其包含治療有效量的式I所示的化合物、其消旋物、立體異構物、互變異構物、同位素標記物、氮氧化物、溶劑化物、多形體、藥學上可接受的鹽或其前藥中的至少一種。The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound of Formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof.

根據本發明的實施方案，所述藥物組合物還包括一種或多種藥學上可接受的輔料。According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

所述藥物組合物中的輔料為「可接受的」，其可與組合物的活性成分相容(並且較佳地，能夠穩定活性成分)並且對被治療的受試者不是有害的。可以使用一種或多種藥物賦形劑用於遞送活性化合物。The excipients in the pharmaceutical compositions are "acceptable" in the sense that they are compatible with the active ingredient of the composition (and preferably, capable of stabilizing the active ingredient) and are not deleterious to the subject being treated. One or more pharmaceutical formulations may be used to deliver the active compound.

根據本發明的一些實施方案，所述藥物組合物還可以進一步含有一種或多種額外的治療劑。According to some embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

本發明進一步提供了式I的化合物、其消旋物、立體異構物、互變異構物、同位素標記物、氮氧化物、溶劑化物、多形體、藥學上可接受的鹽或其前藥或所述藥物組合物在製備藥物中的用途。The present invention further provides the use of the compound of formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof or the pharmaceutical composition in the preparation of drugs.

根據一些實施方案，所述藥物為診斷、預防和/或治療p38α-MK2介導的疾病或病症的藥物。According to some embodiments, the drug is a drug for diagnosing, preventing and/or treating a disease or condition mediated by p38α-MK2.

根據一些實施方案，所述藥物為p38α-MK2抑制劑。According to some embodiments, the drug is a p38α-MK2 inhibitor.

根據一些實施方案，所述p38α-MK2介導的疾病或症狀選自潰瘍性結腸炎、炎症性腸炎、克羅恩病、銀屑病、系統性紅斑狼瘡、多發性硬化症、重症肌無力、骨疾病、骨關節炎、感染性休克、內毒素休克、關節炎、敗血症、哮喘、慢性阻塞性肺病、cryopyrin相關的週期性症候群、類風濕性關節炎、化膿性汗腺炎、僵直性脊椎炎或癌症。According to some embodiments, the p38α-MK2-mediated disease or condition is selected from ulcerative colitis, inflammatory enteritis, Crohn's disease, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, bone disease, osteoarthritis, septic shock, endotoxin shock, arthritis, sepsis, asthma, chronic obstructive pulmonary disease, cryopyrin-related periodic syndrome, rheumatoid arthritis, hidradenitis suppurativa, ankylosing spondylitis or cancer.

根據一些實施方案，可將式I的化合物、其消旋物、立體異構物、互變異構物、同位素標記物、氮氧化物、溶劑化物、多形體、藥學上可接受的鹽或其前藥製成適合於通過任何適當途徑給藥的形式，通過常規方法使用一種或多種藥學上可接受的載體來配製。因此，式I的化合物、其消旋物、立體異構物、互變異構物、同位素標記物、氮氧化物、溶劑化物、多形體、藥學上可接受的鹽或其前藥可以被配製成用於口服給藥、注射(例如靜脈內、肌肉內或皮下)給藥、吸入或吹入給藥的各種劑型；也可以配製成持續釋放劑型，例如片劑、硬或軟膠囊、水性或油性混懸液、乳劑、注射液、可分散性粉末或顆粒、栓劑、錠劑或糖漿。According to some embodiments, the compound of Formula I, its racemate, stereoisomer, tautomer, isotopically labeled, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof can be prepared in a form suitable for administration by any appropriate route and formulated using one or more pharmaceutically acceptable carriers by conventional methods. Therefore, the compounds of formula I, their racemates, stereoisomers, tautomers, isotopically labeled substances, nitrogen oxides, solvates, polymorphs, pharmaceutically acceptable salts or prodrugs thereof can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular or subcutaneous) administration, inhalation or insufflation administration; they can also be formulated into sustained-release dosage forms, such as tablets, hard or soft capsules, aqueous or oily suspensions, emulsions, injections, dispersible powders or granules, suppositories, tablets or syrups.

本發明還提供了一種用於診斷、預防和/或治療p38α-MK2介導的疾病或病症的方法，該方法包括向需要這種治療的患者單獨施用治療有效量的至少一種本發明的化合物或藥物組合物，或任選地，與本發明的另一種化合物和/或至少一種其他類型的治療劑組合。The present invention also provides a method for diagnosing, preventing and/or treating a disease or condition mediated by p38α-MK2, which method comprises administering to a patient in need of such treatment a therapeutically effective amount of at least one compound or pharmaceutical composition of the present invention alone, or, optionally, in combination with another compound of the present invention and/or at least one other type of therapeutic agent.

根據一些實施方案，所述p38α-MK2介導的疾病或症狀選自類風濕性關節炎或僵直性脊椎炎。According to some embodiments, the p38α-MK2-mediated disease or condition is selected from rheumatoid arthritis or ankylosing spondylitis.

在一些實施方案中，所述患者是哺乳動物，較佳是人。In some embodiments, the patient is a mammal, preferably a human.

有益效果(1)本發明提供了一類結構新穎的化合物，其創造性的改變了同類化合物的母核結構，同時仍然取得了良好的p38α-MK2抑制作用。 (2)本發明通過結構優化，創造性的獲得了一類結構新穎的化合物，其不僅具有良好的p38α-MK2抑制作用，同時化合物的選擇性和/或藥代性質得到了提高。 (3)本發明的較佳化合物體內治療效果非常突出，同劑量下顯著優於陽參化合物的治療效果； (4)本發明化合物安全性良好。本發明化合物具有良好的成藥性，可用於治療或預防與p38α-MK2相關的病症和疾病，以及製備用於治療或預防此類病症和疾病的藥物。 Beneficial Effects (1) The present invention provides a class of compounds with novel structures, which creatively change the parent core structure of similar compounds, while still achieving good p38α-MK2 inhibitory effects. (2) The present invention creatively obtains a class of compounds with novel structures through structural optimization, which not only have good p38α-MK2 inhibitory effects, but also have improved selectivity and/or pharmacokinetic properties. (3) The preferred compounds of the present invention have very outstanding in vivo therapeutic effects, which are significantly superior to the therapeutic effects of Yangshen compounds at the same dosage; (4) The compounds of the present invention have good safety. The compounds of the present invention have good drugability and can be used to treat or prevent p38α-MK2-related diseases and disorders, as well as to prepare drugs for treating or preventing such diseases and disorders.

用語定義與說明Definitions and Explanations

除非另有說明，本發明說明書和申請專利範圍中記載的基團和用語定義，包括其作為實例的定義、示例性的定義、較佳的定義、表格中記載的定義、實施例中具體化合物的定義等，可以彼此之間任意組合和結合。這樣的組合和結合後的基團定義及化合物結構，應當被理解為本發明說明書和/或申請專利範圍記載的範圍內。Unless otherwise stated, the definitions of groups and terms described in the present invention specification and the scope of the patent application, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined and combined with each other. The group definitions and compound structures after such combination and combination should be understood to be within the scope described in the present invention specification and/or the scope of the patent application.

本發明通式定義中的用語「任選的」(或「任選地」、「任選」)意味著被零個、一個或多個取代基所取代的情形，例如「任選被一個、兩個或更多個R取代」意味著可以不被R取代(無取代)或可以選擇被一個、兩個或更多個R取代。The term "optionally" (or "optionally", "optionally") in the general formula definition of the present invention means substituted by zero, one or more substituents, for example, "optionally substituted by one, two or more R" means that it may not be substituted by R (unsubstituted) or may be selectively substituted by one, two or more R.

「更多個」表示三個或三個以上，例如3、4、5、6、7、8、9或10。“More” means three or more, such as 3, 4, 5, 6, 7, 8, 9 or 10.

除非另有說明，本說明書和申請專利範圍記載的數值範圍相當於至少記載了其中每一個具體的整數數值。例如，數值範圍「1-12」相當於記載了數值範圍「1-12」中的每一個整數數值，即1、2、3、4、5、6、7、8、9、10、11、12。Unless otherwise stated, the numerical ranges described in this specification and patent application are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to describing each integer value in the numerical range "1-12", namely, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

用語「鹵素」表示氟、氯、溴和碘。The term "halogen" refers to fluorine, chlorine, bromine and iodine.

「HO-C _1-6烷基」指代羥基取代的C _1-6的烷基。 "HO-C _1-6 alkyl" refers to a C _1-6 alkyl group substituted with a hydroxy group.

用語「C _1-6烷基」表示具有1、2、3、4、5或6個碳原子的直鏈和支鏈烷基。所述烷基是例如甲基、乙基、丙基、丁基、戊基、己基、異丙基、異丁基、仲丁基、叔丁基、異戊基、2-甲基丁基、1-甲基丁基、1-乙基丙基、1,2-二甲基丙基、新戊基、1,1-二甲基丙基、4-甲基戊基、3-甲基戊基、2-甲基戊基、1-甲基戊基、2-乙基丁基、1-乙基丁基、3,3-二甲基丁基、2,2-二甲基丁基、1,1-二甲基丁基、2,3-二甲基丁基、1,3-二甲基丁基或1,2-二甲基丁基等或它們的異構物。 The term "C _1-6 alkyl" refers to straight and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl or the like or isomers thereof.

用語「C _3-8環烷基」應理解為表示飽和的一價單環、雙環(如橋環、螺環)烴環，其具有3、4、5、6、7、8個碳原子。所述C _3-8環烷基可以是單環烴基，如環丙基、環丁基、環戊基、環己基、環庚基、環辛基、環壬基或環癸基，或者是雙環烴基如龍腦基、吲哚基、六氫吲哚基、四氫萘基、十氫萘基、二環[2.1.1]己基、二環[2.2.1]庚基、二環[2.2.1]庚烯基、6,6-二甲基二環[3.1.1]庚基、2,6,6-三甲基二環[3.1.1]庚基、二環[2.2.2]辛基、2,7-二氮雜螺[3,5]壬烷基、2,6-二氮雜螺[3,4]辛烷基。 The term "C _3-8 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (eg bridged, spiro) hydrocarbon ring having 3, 4, 5, 6, 7, or 8 carbon atoms. The _C3-8 cycloalkyl group may be a monocyclic alkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic alkyl group such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonanyl, 2,6-diazaspiro[3,4]octanyl.

用語「3-10員雜環基」意指飽和的或不飽和的非芳族的環或環系，並且含有至少一個選自O、S和N的雜原子。所述雜環基可以通過所述碳原子中的任一個或氮原子(如果存在的話)與分子的其餘部分連接。所述雜環基可以包括稠合的或橋連的環以及螺環的環。特別地，所述雜環基可以包括但不限於：4員環，如氮雜環丁烷基、氧雜環丁烷基；5員環，如四氫呋喃基、二氧雜環戊烯基、吡咯烷基、咪唑烷基、吡唑烷基、吡咯啉基；或6員環，如四氫吡喃基、哌啶基、嗎啉基、二噻烷基、硫代嗎啉基、哌嗪基或三噻烷基；或7員環，如二氮雜環庚烷基。任選地，所述雜環基可以是苯並稠合的。所述雜環基可以是雙環的，例如但不限於5,5員環，如六氫環戊並[c]吡咯-2(1H)-基環，或者5,6員雙環，如六氫吡咯並[1,2-a]吡嗪-2(1H)-基環。雜環基可以是部分不飽和的，即它可以包含一個或多個雙鍵，例如但不限於二氫呋喃基、二氫吡喃基、2,5-二氫-1H-吡咯基、4H-[1,3,4]噻二嗪基、1,2,3,5-四氫噁唑基或4H-[1,4]噻嗪基，或者，它可以是苯並稠合的，例如但不限於二氫異喹啉基。所述3-10員雜環基與其它基團相連構成本發明的化合物時，可以為3-10員雜環基上的碳原子與其它基團相連，也可以為3-10員雜環基環上雜環原子與其它基團相連。例如當3-10員雜環基選自哌嗪基時，可以為哌嗪基上的氮原子與其它基團相連。或當3-10員雜環基選自哌啶基時，可以為哌啶基環上的氮原子和其對位上的碳原子與其它基團相連。The term "3-10 membered heterocyclic group" means a saturated or unsaturated non-aromatic ring or ring system and contains at least one heteroatom selected from O, S and N. The heterocyclic group may be attached to the rest of the molecule through any of the carbon atoms or the nitrogen atom (if present). The heterocyclic group may include fused or bridged rings as well as spirocyclic rings. In particular, the heterocyclic group may include, but is not limited to: a 4-membered ring, such as azacyclobutane, oxacyclobutane; a 5-membered ring, such as a tetrahydrofuranyl, a dioxacyclopentane, a pyrrolidinyl, an imidazolidinyl, a pyrazolidinyl, a pyrrolinyl; or a 6-membered ring, such as a tetrahydropyranyl, a piperidinyl, a morpholinyl, a dithianyl, a thiomorpholinyl, a piperazinyl or a trithianyl; or a 7-membered ring, such as a diazacycloheptanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, for example but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring. The heterocyclic group may be partially unsaturated, i.e. it may contain one or more double bonds, for example but not limited to dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, for example but not limited to dihydroisoquinolinyl. When the 3-10 membered heterocyclic group is connected with other groups to form the compound of the present invention, the carbon atom on the 3-10 membered heterocyclic group may be connected with the other groups, or the heterocyclic atom on the 3-10 membered heterocyclic group may be connected with the other groups. For example, when the 3-10 membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl may be connected with the other groups. Or when the 3-10 membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and the carbon atom at the para position thereof may be connected with the other groups.

用語「5-10員雜芳基」應理解為包括這樣的一價單環或雙環環系：其具有5、6、7、8、9或10個環原子，且其包含1-5個，較佳為1-3個獨立選自N、O和S的雜原子並且，另外在每一種情況下可為苯並稠合的。單環「雜芳基」的實例包括例如吡啶基、噠嗪基、嘧啶基、吡嗪基、噻嗪基、噁嗪基、三嗪基、噻二嗪基或噁二嗪基等。「雜芳基」還指其中雜芳族環與一個或多個芳基、脂環族或雜環基環稠合的基團，其中所述連接的位點在雜芳族環上。非限制性實例包括1-、2-、3-、5-、6-、7-或8-吲嗪基、1-、3-、4-、5-、6-或7-異吲哚基、2-、3-、4-、5-、6-或7-吲哚基、2-、3-、4-、5-、6-或7-吲唑基、2-、4-、5-、6-、7-或8-嘌呤基、1-、2-、3-、4-、6-、7-、8-或9-喹嗪基、2-、3-、4-、5-、6-、7-或8-喹啉基、1-、3-、4-、5-、6-、7-或8-異喹啉基、1-、4-、5-、6-、7-或8-酞嗪基(phthalazinyl)、2-、3-、4-、5-或6-萘啶基、2-、3-、5-、6-、7-或8-喹唑啉基、3-、4-、5-、6-、7-或8-噌啉基、2-、4-、6-或7-蝶啶基、1-、2-、3-、4-、5-、6-、7-或8-4aH咔唑基、1-、2-、3-、4-、5-、6-、7-或8-咔唑基、1-、3-、4-、5-、6-、7-、8-或9-咔啉基、1-、2-、3-、4-、6-、7-、8-、9-或10-菲啶基、1-、2-、3-、4-、5-、6-、7-、8-或9-吖啶基、1-、2-、4-、5-、6-、7-、8-或9-啶基、2-、3-、4-、5-、6-、8-、9-或10-菲咯啉基、1-、2-、3-、4-、6-、7-、8-或9-吩嗪基、1-、2-、3-、4-、6-、7-、8-、9-或10-吩噻嗪基、1-、2-、3-、4-、6-、7-、8-、9-或10-吩嗪基、2-、3-、4-、5-、6-或1-、3-、4-、5-、6-、7-、8-、9-或10-苯並異喹啉基、2-、3-、4-或噻吩並[2,3-b]呋喃基、2-、3-、5-、6-、7-、8-、9-、10-或11-7H-吡嗪並[2,3-c]咔唑基、2-、3-、5-、6-或7-2H-呋喃並[3,2-b]-吡喃基、2-、3-、4-、5-、7-或8-5H-吡啶並[2,3-d]-鄰-嗪基、1-、3-或5-1H-吡唑並[4,3-d]-唑基、2-、4-或54H-咪唑並[4,5-d]噻唑基、3-、5-或8-吡嗪並[2,3-d]噠嗪基、2-、3-、5-或6-咪唑並[2,1-b]噻唑基、1-、3-、6-、7-、8-或9-呋喃並[3,4-c]噌啉基、1-、2-、3-、4-、5-、6-、8-、9-、10或11-4H-吡啶並[2,3-c]咔唑基、2-、3-、6-或7-咪唑並[1,2-b][1,2,4]三嗪基、7-苯並[b]噻吩基、2-、4-、5-、6-或7-苯並唑基、2-、4-、5-、6-或7-苯並咪唑基、2-、4-、4-、5-、6-或7-苯並噻唑基、1-、2-、4-、5-、6-、7-、8-或9-苯並氧雜基(benzoxapinyl)、2-、4-、5-、6-、7-或8-苯並嗪基、1-、2-、3-、5-、6-、7-、8-、9-、10-或11-4H-吡咯並[1,2-b][2]苯並氮雜基(benzazapinyl)。典型的稠合雜芳基包括但不限於2-、3-、4-、5-、6-、7-或8-喹啉基、1-、3-、4-、5-、6-、7-或8-異喹啉基、2-、3-、4-、5-、6-或7-吲哚基、2-、3-、4-、5-、6-或7-苯並[b]噻吩基、2-、4-、5-、6-或7-苯並唑基、2-、4-、5-、6-或7-苯並咪唑基和2-、4-、5-、6-或7-苯並噻唑基。當所述5-10員雜芳基與其它基團相連構成本發明的化合物時，可以為5-10員雜芳基環上的碳原子與其它基團相連，也可以為5-10員雜芳基環上的雜原子與其它基團相連。當所述5-10員雜芳基被取代時，其可以為單取代或者多取代。並且，對其取代位點沒有限制，例如可以為雜芳基環上與碳原子相連的氫被取代，或者雜芳基環上與雜原子相連的氫被取代。The term "5-10 membered heteroaryl" is to be understood as including monovalent monocyclic or bicyclic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms and which contain 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S and which, in addition, in each case may be benzofused. Examples of monocyclic "heteroaryl" include, for example, pyridyl, oxazinyl, pyrimidinyl, pyrazinyl, thiazinyl, oxazinyl, triazinyl, thiadiazinyl or oxadiazinyl, etc. "Heteroaryl" also refers to radicals in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclic rings, wherein the point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 1-, 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-carbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-acryl. 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenylindole, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenylindole, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenylindole, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenylindole, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenylindole, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenylindole, 1-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-benzoisoquinolyl, 2-, 3-, 4-, or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6-, or 7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7-, or 8-5H-pyrido[2,3-d]-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]oxazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9- furano[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6- or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7- or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-10 membered heteroaryl group is linked to other groups to form the compound of the present invention, the carbon atoms on the 5-10 membered heteroaryl ring may be linked to the other groups, or the hetero atoms on the 5-10 membered heteroaryl ring may be linked to the other groups. When the 5-10 membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution position, for example, a hydrogen atom connected to a carbon atom on the heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on the heteroaryl ring may be substituted.

用語「氮氧化物」是指叔胺類或含氮(芳)雜環類化合物結構中的氮原子經氧化而形成的化合物。The term "nitrogen oxides" refers to compounds formed by oxidation of nitrogen atoms in the structures of tertiary amines or nitrogen-containing (aromatic) heterocyclic compounds.

用語「螺環」是指兩個環共用1個成環原子的環系。The term "spiro" refers to a ring system in which two rings share one ring atom.

用語「稠環」是指兩個環共用2個成環原子的環系。The term "fused ring" refers to a ring system in which two rings share two ring atoms.

用語「橋環」是指兩個環共用3個以上成環原子的環系。The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

除非另有說明，雜環基、雜芳基或亞雜芳基包括其所有可能的異構形式，例如其位置異構物。因此，對於一些說明性的非限制性實例，可以包括在其1-、2-、3-、4-、5-、6-、7-、8-、9-、10-、11-、12-位等(如果存在)中的1、2個或更多個位置上取代或與其他基團鍵合的形式，包括吡啶-2-基、亞吡啶-2-基、吡啶-3-基、亞吡啶-3-基、吡啶-4-基和亞吡啶-4-基；噻吩基或亞噻吩基包括噻吩-2-基、亞噻吩-2-基、噻吩-3-基和亞噻吩-3-基；吡唑-1-基、吡唑-3-基、吡唑-4-基、吡唑-5-基。Unless otherwise specified, a heterocyclic group, heteroaryl group or heteroarylene group includes all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, forms substituted or bonded to other groups at 1, 2 or more positions of the 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) may include pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-ylene and pyridin-4-ylene; thienyl or thienylene group includes thien-2-yl, thien-2-ylene, thien-3-ylene and thien-3-ylene; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl.

用語「側氧基」是指取代基中的碳原子、氮原子或硫原子被氧化後形成的氧基取代(=O)。The term "oxo group" refers to an oxy group (=O) formed by oxidation of a carbon atom, a nitrogen atom or a sulfur atom in a substituent.

用語「烷基胺基」指-NH-(烷基)或-N-(烷基) ₂，其中烷基的定義如上所述。烷基胺基的非限制性實例包括：甲胺基、乙基胺基、丙基胺基、異丙基胺基、丁基胺基、二甲基胺基、甲基乙基胺基、二乙基胺基、二丙基胺基、甲基丙基胺基、二異丙基胺基、二丁基胺基等。 The term "alkylamino" refers to -NH-(alkyl) or -N-(alkyl) ₂ , wherein alkyl is as defined above. Non-limiting examples of alkylamino include methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, and the like.

用語「烷基氧基」指-O-(烷基)，其中烷基的定義如上所述。烷氧基的非限制性實例包括：甲氧基、乙氧基、丙氧基、丁氧基。烷氧基可以是任選取代的或非取代的，當被取代時，取代基較佳為一個或多個以下基團，其獨立地選自烷基、烯基、炔基、烷基氧基、烷基胺基、鹵素、巰基、羥基、硝基、腈基、環烷基、雜環烷基、芳基、雜芳基、環烷基氧基或雜環烷基氧基。The term "alkyloxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, hydroxyl, nitro, nitrile, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy or heterocycloalkyloxy.

用語「亞烷基氧基」和「氧基亞烷基」指-亞烷基-O-或-O-亞烷基-，其中的亞烷基表示直鏈或支鏈飽和二價烴基。關於「亞烷基」的碳原子數的定義適用上文對「烷基」的定義。本發明所屬技術領域中具有通常知識者能夠理解，亞烷基氧基或氧基亞烷基可以以任意方向與包含它的分子的其餘部分連接，即二者可以互換地使用。The terms "alkyleneoxy" and "oxyalkylene" refer to -alkylene-O- or -O-alkylene-, wherein alkylene represents a linear or branched saturated divalent hydrocarbon group. The definition of the number of carbon atoms in "alkylene" is applicable to the definition of "alkyl" above. Those skilled in the art to which the present invention belongs can understand that alkyleneoxy or oxyalkylene can be connected to the rest of the molecule containing it in any direction, that is, the two can be used interchangeably.

「鹵代烷基」指被一個或多個鹵素取代的烷基，其中烷基如上所定義。"Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

帶有符號「#」標記的化合物表示該化合物為軸掌性化合物，本領域公知，軸掌性化合物為含有掌性軸的立體異構物化合物，其通常以Ra/Sa或P/M標記，即帶有符號「#」標記的化合物可能為P或Ra構型軸掌性化合物，也可能為M或Sa構型軸掌性化合物；帶有符號「＊」標記的碳原子為掌性碳原子，每個掌性碳原子可以基於立體化學而被定義為(R)‑或(S)‑，即帶有符號「＊」標記的碳原子可能為R構型，也可能為S構型。A compound marked with the symbol "#" indicates that the compound is an axial chiral compound. It is well known in the art that an axial chiral compound is a stereoisomer compound containing a chiral axis, which is usually marked with Ra/Sa or P/M, that is, a compound marked with the symbol "#" may be an axial chiral compound with a P or Ra configuration, or an axial chiral compound with an M or Sa configuration; a carbon atom marked with the symbol "*" is a chiral carbon atom, and each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry, that is, a carbon atom marked with the symbol "*" may be an R configuration or an S configuration.

本發明所述化合物的化學結構中，鍵「」表示未指定構型，「」或「」表示絕對構型，即如果化學結構中存在立體異構物，鍵「」可以為「」或「」，或者同時包含「」和「」兩種構型；「」表示存在軸掌性。如化合物「」和「」即表示一對軸掌性異構物，本發明中其他化學結構也作相同解釋。 In the chemical structure of the compound of the present invention, the bond " " indicates that the configuration is not specified, " "or" " indicates the absolute configuration, that is, if stereoisomers exist in the chemical structure, the key " " can be " "or" ", or both "and" "Two configurations;" " indicates the presence of axial chirality. For example, the compound " "and" ” means a pair of axial chiral isomers, and other chemical structures in the present invention shall be interpreted in the same manner.

在本發明中，所涉及的化合物亦包括經同位素標記的化合物，所述經同位素標記的化合物與式I中所示的那些相同，但是其中一或多個原子被原子重量或質量數不同於通常天然存在的原子重量或質量數的原子替代。可摻入本發明的化合物的同位素的實例包括H、C、N、O、S、F及Cl的同位素，分別諸如 ²H、 ³H、 ¹³C、 ¹¹C、 ¹⁴C、 ¹⁵N、 ¹⁸O、 ¹⁷O、 ³²P、 ³⁵S、 ¹⁸F及 ³⁶Cl。含有上述同位素和/或其他原子的其他同位素的本發明的化合物、其前藥、或者所述化合物或所述前藥的藥學上可接受的鹽在本發明的範圍內。本發明的某些經同位素標記的化合物，例如摻入放射性同位素(諸如 ³H和 ¹⁴C)的化合物可用於藥物和/或受質組織分佈測定。氚(即 ³H)和碳14(即 ¹⁴C)同位素因易於製備和可檢測性而成為特別佳的。再者，以較重的同位素(諸如氘，即 ²H或D)替代可提供源自更高的代謝穩定性的某些治療優勢(例如增加的體內半衰期或減少的劑量需求)，並因此可在某些情況下是較佳的。如請求項所請求保護的本發明化合物可特別地限定以氘或氚替代。此外，取代基中出現的氫未單獨列明用語氘或氚並不表示排除氘或氚，而是同樣也可以包含氘或氚。 In the present invention, the compounds involved also include isotopically labeled compounds, which are the same as those shown in Formula I, but one or more atoms are replaced by atoms whose atomic weight or mass number is different from the atomic weight or mass number usually occurring in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F and Cl, such as ² H, ³ H, ¹³ C, ¹¹ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl, respectively. The compounds of the present invention, their prodrugs, or pharmaceutically acceptable salts of the compounds or the prodrugs containing the above-mentioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Certain isotopically labeled compounds of the invention, for example compounds incorporating radioactive isotopes such as ³ H and ¹⁴ C, are useful in drug and/or substrate tissue distribution assays. Tritium (i.e., ³ H) and carbon 14 (i.e., ¹⁴ C) isotopes are particularly preferred due to ease of preparation and detectability. Furthermore, substitution with heavier isotopes such as deuterium, i.e., ² H or D, may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements), and therefore may be preferred in certain circumstances. The compounds of the invention as claimed in the claims may be specifically limited to substitution with deuterium or tritium. Furthermore, the presence of hydrogen in a substituent without the separate listing of the term deuterium or tritium does not exclude deuterium or tritium, but may also include deuterium or tritium.

本發明所屬技術領域中具有通常知識者可以理解，式(I)所示化合物可以以各種藥學上可接受的鹽的形式存在。如果這些化合物具有鹼性中心，則其可以形成酸加成鹽；如果這些化合物具有酸性中心，則其可以形成鹼加成鹽；如果這些化合物既包含酸性中心(例如羧基)又包含鹼性中心(例如胺基)，則其還可以形成內鹽。Those skilled in the art will appreciate that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form an acid addition salt; if these compounds have an acidic center, they may form an alkali addition salt; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amine group), they may also form an internal salt.

本發明的化合物可以溶劑合物(如水合物)的形式存在，其中本發明的化合物包含作為所述化合物晶格的結構要素的極性溶劑，特別是例如水、甲醇或乙醇。極性溶劑特別是水的量可以化學計量比或非化學計量比存在。The compounds of the invention may exist in the form of solvates (e.g. hydrates), wherein the compounds of the invention contain polar solvents as structural elements of the crystal lattice of the compounds, in particular water, methanol or ethanol, for example. The polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric amount.

根據其分子結構，本發明的化合物可以是掌性的，因此可能存在各種鏡像異構物形式。因而這些化合物可以以消旋物形式或光學活性形式存在。本發明的化合物涵蓋了各掌性碳為R或S構型的異構物或其混合物、消旋物。本發明的化合物或其中間體可以通過本發明所屬技術領域中具有通常知識者公知的化學或物理方法分離為鏡像異構物化合物，或者以此形式用於合成。在外消旋的胺的情況中，通過與光學活性的拆分試劑反應，從混合物製得非鏡像異構物。適當的拆分試劑的示例是光學活性的酸，例如R和S形式的酒石酸、二乙醯酒石酸、二苯甲醯酒石酸、扁桃酸、蘋果酸、乳酸、適當的N-保護的胺基酸(例如N-苯甲醯脯胺酸或N-苯磺醯基脯胺酸)或各種光學活性的樟腦磺酸。借助光學活性的拆分試劑(例如固定在矽膠上的二硝基苯甲醯基苯基甘胺酸、三乙酸纖維素或其它碳水化合物的衍生物或掌性衍生化的異丁烯酸酯聚合物)，也可有利地進行色譜鏡像物拆分。用於此目的的適當的洗脫劑是含水或含醇的溶劑混合物，例如，己烷/異丙醇/乙腈。According to their molecular structure, the compounds of the present invention may be chiral, and therefore may exist in various mirror image isomer forms. Therefore, these compounds may exist in the form of racemates or optically active forms. The compounds of the present invention cover isomers or mixtures and racemates thereof in which each chiral carbon is in R or S configuration. The compounds of the present invention or their intermediates can be separated into mirror image isomer compounds by chemical or physical methods known to those of ordinary skill in the art to which the present invention belongs, or used in this form for synthesis. In the case of racemic amines, non-mirror image isomers are prepared from the mixture by reaction with an optically active resolution agent. Examples of suitable resolution agents are optically active acids, such as tartaric acid in its R and S forms, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, apple acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids. Chromatographic image resolution can also be advantageously performed with the aid of optically active resolution agents (e.g. dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, e.g. hexane/isopropanol/acetonitrile.

可以根據已知的方法，例如通過萃取、過濾或柱層析來分離相應的穩定異構物。The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column analysis.

用語「患者」是指包括哺乳動物在內的任何動物，較佳為小鼠、大鼠、其它齧齒類動物、兔、狗、貓、豬、牛、羊、馬或靈長類動物，最佳為人。The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

用語「治療有效量」是指研究人員、獸醫、醫師或其它臨床醫師正在組織、系統、動物、個體或人中尋找的引起生物學或醫學反應的活性化合物或藥物的量，它包括以下一項或多項：(1)預防疾病：例如在易感染疾病、紊亂或病症但尚未經歷或出現疾病病理或症狀的個體中預防疾病、紊亂或病症。(2)抑制疾病：例如在正經歷或出現疾病、紊亂或病症的病理或症狀的個體中抑制疾病、紊亂或病症(即阻止病理和/或症狀的進一步發展)。(3)緩解疾病：例如在正經歷或出現疾病、紊亂或病症的病理或症狀的個體中緩解疾病、紊亂或病症(即逆轉病理和/或症狀)。The term "therapeutically effective amount" refers to the amount of an active compound or drug that elicits the biological or medicinal response that a researcher, veterinarian, physician or other clinician is seeking in a tissue, system, animal, individual or human, and includes one or more of the following: (1) Preventing disease: e.g., preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed the pathology or symptoms of the disease. (2) Inhibiting disease: e.g., inhibiting the disease, disorder or condition (i.e., preventing further development of the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition. (3) Relieving disease: e.g., relieving the disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition.

具體實施方式Specific implementation methods

下文將結合具體實施例對本發明的技術方案做更進一步的詳細說明。應當理解，下列實施例僅為示例性地說明和解釋本發明，而不應被解釋為對本發明保護範圍的限制。凡基於本發明上述內容所實現的技術均涵蓋在本發明旨在保護的範圍內。The following will further explain the technical solution of the present invention in combination with specific embodiments. It should be understood that the following embodiments are only for illustrative purposes to illustrate and explain the present invention, and should not be interpreted as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope of protection intended by the present invention.

除非另有說明，以下實施例中使用的原料和試劑均為市售商品，或者可以通過已知方法製備。Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

本發明的化合物結構是通過核磁共振(NMR)或/和液質聯用管柱層析(LC-MS)來確定的。NMR化學位移(δ)以百萬分之一(ppm)的單位給出。NMR的測定是用Bruker AVANCE-400核磁儀，測定溶劑為氘代二甲基亞碸(DMSO- d ₆ )，氘代甲醇(CD ₃OD)和氘代氯仿(CDCl ₃)，內標為四甲基矽烷(TMS)。 The structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass spectrometry (LC-MS). The NMR chemical shift (δ) is given in parts per million (ppm). The NMR measurement is performed using a Bruker AVANCE-400 nuclear magnetic spectrometer, and the measurement solvents are deuterated dimethyl sulfoxide (DMSO -d ₆ ), deuterated methanol (CD ₃ OD) and deuterated chloroform (CDCl ₃ ), and the internal standard is tetramethylsilane (TMS).

液質聯用管柱層析LC-MS的測定用Agilent 1200 Infinity Series質譜儀。HPLC的測定使用安捷倫1200DAD高壓液相管柱層析儀(Sunfire C18 150×4.6 mm管柱層析管柱)和Waters 2695-2996高壓液相管柱層析儀(Gimini C18 150×4.6mm管柱層析管柱)。Liquid chromatography-mass spectrometry (LC-MS) was performed on an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed on an Agilent 1200DAD high pressure liquid chromatography column (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatography column (Gimini C18 150×4.6 mm column).

薄層層析矽膠板使用煙臺黃海HSGF254或青島GF254矽膠板，TLC採用的規格是0.15 mm～0.20 mm，薄層層析分離純化產品採用的規格是0.4 mm～0.5 mm。柱層析一般使用煙臺黃海矽膠200～300目矽膠為載體。Thin layer chromatography silica plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica plates. The specifications used for TLC are 0.15 mm to 0.20 mm, and the specifications used for thin layer chromatography separation and purification products are 0.4 mm to 0.5 mm. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

在無特殊說明的情況下，本發明的所有反應均在連續的磁力攪拌下，在乾燥氮氣或氬氣氛下進行，溶劑為乾燥溶劑，反應溫度單位為攝氏度。Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, the solvent is a dry solvent, and the reaction temperature is in degrees Celsius.

實施例1Embodiment 1

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙醯胺 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropionamide

第一步first step

2,2-二甲基-6-(2-氧丙基)-4H-1,3-二噁英-4-酮的製備Preparation of 2,2-dimethyl-6-(2-oxypropyl)-4H-1,3-dioxin-4-one

氮氣保護下，向2,2,6-三甲基-4H-1,3-二英-4-酮Cpd-01a(30 g, 211 mmol)的四氫呋喃(200 mL)溶液中滴加雙(三甲基矽基)胺基鋰的四氫呋喃溶液(1.0 M; 500 mL, 500 mmol)。反應混合物在-20℃中攪拌4小時後，滴加二乙基鋅的四氫呋喃溶液(1.0 M; 500 mL, 500 mmol)。4小時後，在-10℃加入N-乙醯基咪唑(32.4 g, 294 mmol)。反應混合物攪拌16小時，緩慢恢復室溫。反應結束後，旋乾四氫呋喃溶液，用6M鹽酸調節pH為2，用乙酸乙酯(3 × 10 mL)萃取，無水硫酸鈉乾燥有機相，濃縮濾液後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=7/1)純化後得到2,2-二甲基-6-(2-氧丙基)-4H-1,3-二噁英-4-酮Cpd-01b(25 g)，產率：64%。Under nitrogen protection, a solution of bis(trimethylsilyl)amide lithium in tetrahydrofuran (1.0 M; 500 mL, 500 mmol) was added dropwise to a solution of 2,2,6-trimethyl-4H-1,3-dioxin-4-one Cpd-01a (30 g, 211 mmol) in tetrahydrofuran (200 mL). The reaction mixture was stirred at -20°C for 4 hours, and then a solution of diethylzinc in tetrahydrofuran (1.0 M; 500 mL, 500 mmol) was added dropwise. After 4 hours, N-acetylimidazole (32.4 g, 294 mmol) was added at -10°C. The reaction mixture was stirred for 16 hours and slowly returned to room temperature. After the reaction, the tetrahydrofuran solution was dried by rotary evaporation, the pH was adjusted to 2 with 6 M hydrochloric acid, extracted with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 7/1) to obtain 2,2-dimethyl-6-(2-oxopropyl)-4H-1,3-dioxin-4-one Cpd-01b (25 g) with a yield of 64%.

MS m/z(ESI)：185.2(M+1).MS m/z(ESI):185.2(M+1).

第二步Step 2

2'-氯-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 2'-chloro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

將2,2-二甲基-6-(2-氧丙基)-4H-1,3-二噁英-4-酮Cpd-01b(25 g，135 mmol)和2-氯-4-胺基-5-甲基吡啶(29 g，203 mmol)溶於1,4-二氧六環(250 mL)。反應混合物在90℃攪拌3.5小時後，滴加濃硫酸(12.5 mL)接著反應1小時。反應結束後，濃縮，向反應容器中加入水(200 mL)，攪拌30分鐘。反應混合物過濾得到2'-氯-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01c(30 g)，產率：88%。2,2-Dimethyl-6-(2-oxopropyl)-4H-1,3-dioxin-4-one Cpd-01b (25 g, 135 mmol) and 2-chloro-4-amino-5-methylpyridine (29 g, 203 mmol) were dissolved in 1,4-dioxane (250 mL). The reaction mixture was stirred at 90°C for 3.5 hours, and then concentrated sulfuric acid (12.5 mL) was added dropwise and the reaction was continued for 1 hour. After the reaction was completed, the mixture was concentrated, and water (200 mL) was added to the reaction vessel and stirred for 30 minutes. The reaction mixture was filtered to obtain 2'-chloro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01c (30 g) with a yield of 88%.

MS m/z(ESI)：251.0(M+1)。MS m/z (ESI): 251.0 (M+1).

第三步third step

2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

將2'-氯-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01c(20 g, 0.0798 mol)溶於N,N-二甲基甲醯胺(200 mL)中，加入2-(溴甲基)-3,5-二氟吡啶(33.2 g, 0.1596 mol)、碳酸鉀(33.09 g, 0.2394 mol)、18-冠醚-6(2.11 g, 0.0080 mol)，氮氣下於60℃反應16小時。反應結束後，加入水，乙酸乙酯萃取，收集有機相、乾燥、過濾、富集。殘液經矽膠管柱(石油醚/乙酸乙酯=2/1)得到2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01d(18 g)，產率：59.7%。2'-Chloro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01c (20 g, 0.0798 mol) was dissolved in N,N-dimethylformamide (200 mL), and 2-(bromomethyl)-3,5-difluoropyridine (33.2 g, 0.1596 mol), potassium carbonate (33.09 g, 0.2394 mol), and 18-crown-6 (2.11 g, 0.0080 mol) were added, and the mixture was reacted at 60°C for 16 hours under nitrogen. After the reaction was completed, water was added, and the mixture was extracted with ethyl acetate. The organic phase was collected, dried, filtered, and enriched. The residue was purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to give 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-01d (18 g) with a yield of 59.7%.

MS m/z(ESI)：378.7(M+1)。MS m/z(ESI):378.7(M+1).

第四步the fourth step

2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 2'-acetyl-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

將2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01d(18 g, 0.0476 mol)溶於1,4-二氧六環(200 mL)中，加入三丁基(1-乙氧基乙烯)錫(18.91 g, 0.0524 mol)，雙三苯基磷二氯化鈀(3.34 g, 0.0048 mol)，於120℃反應16小時。反應結束後，過濾、旋乾。繼續將殘液溶於四氫呋喃(200 mL)中，加入濃鹽酸(9 mL)，室溫反應1小時。反應結束後，旋乾，殘餘物經矽膠管柱(石油醚/乙酸乙酯=2/1)得到2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01e(18 g)，產率：98.1%。Dissolve 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01d (18 g, 0.0476 mol) in 1,4-dioxane (200 mL), add tributyl(1-ethoxyethylene)tin (18.91 g, 0.0524 mol), bis(triphenylphosphine)palladium dichloride (3.34 g, 0.0048 mol), and react at 120°C for 16 hours. After the reaction is completed, filter and spin dry. Dissolve the residue in tetrahydrofuran (200 mL), add concentrated hydrochloric acid (9 mL), and react at room temperature for 1 hour. After the reaction was completed, the reaction mixture was dried by rotary evaporation, and the residue was purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 2'-acetyl-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01e (18 g) with a yield of 98.1%.

MS m/z(ESI)：386.7(M+1)。MS m/z(ESI):386.7(M+1).

第五步the fifth step

2'-乙醯基-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 2'-acetyl-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

將2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01e(18 g, 0.0467 mol)溶於異丙醇(200 mL)中，加入N-氯代丁二醯亞胺(6.86 g, 0.0514 mol)，二氯乙酸(0.6 g, 0.0047 mol)，於60℃反應3小時。反應結束後，旋乾溶劑，殘餘物經矽膠管柱純化(石油醚/乙酸乙酯=2/1)得到2'-乙醯基-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01f(13 g)，產率：66.4%。2'-Acetyl-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01e (18 g, 0.0467 mol) was dissolved in isopropanol (200 mL), N-chlorosuccinimide (6.86 g, 0.0514 mol) and dichloroacetic acid (0.6 g, 0.0047 mol) were added, and the mixture was reacted at 60°C for 3 hours. After the reaction, the solvent was dried by rotary evaporation, and the residue was purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 2'-acetyl-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01f (13 g) with a yield of 66.4%.

MS m/z(ESI)：420.7(M+1)。MS m/z(ESI):420.7(M+1).

第六步Step 6

(E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲胺基)丙烯醯基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of (E)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

將2'-乙醯基-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01f(300 mg, 0.714 mmol)溶於N,N-二甲基甲醯胺(DMF, 5 mL)，隨後加入DMF-DMA(340 mg, 2.86 mmol)，反應在55℃下攪拌16小時後，旋蒸除去DMF，得到粗產品(E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲胺基)丙烯醯基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01g(300 mg)。2'-Acetyl-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-01f (300 mg, 0.714 mmol) was dissolved in N,N-dimethylformamide (DMF, 5 mL), and then DMF-DMA (340 mg, 2.86 mmol) was added. The reaction was stirred at 55°C for 16 hours, and then DMF was removed by rotary evaporation to obtain the crude product (E)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-01g (300 mg).

MS m/z(ESI)：475.2(M+1)。MS m/z(ESI):475.2(M+1).

第七步Step 7

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸乙酯的製備Preparation of ethyl 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanoate

將3-胺基-3-亞胺基-2,2-二甲基丙酸乙酯Cpd-01m(1.5 g, 9 mmol)溶於DMF(5 mL)，加入(E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲胺基)丙烯醯基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-01g(300 mg, 0.63 mmol)以及碳酸鉀(174 mg, 1.26 mmol)，反應液在75℃下攪拌4小時，反應結束後，抽濾、旋乾，濃縮後經矽膠管柱(二氯甲烷/甲醇=30/1)分離純化得到2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸乙酯Cpd-01h(150 mg)，收率42%。3-Amino-3-imino-2,2-dimethylpropionic acid ethyl ester Cpd-01m (1.5 g, 9 mmol) was dissolved in DMF (5 mL), and (E)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-01g (300 mg, 0.63 mmol) and potassium carbonate (174 mg, 1.26 mmol), the reaction solution was stirred at 75°C for 4 hours. After the reaction was completed, it was filtered and dried by rotation. After concentration, it was separated and purified by silica gel column (dichloromethane/methanol=30/1) to obtain 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropionic acid ethyl ester Cpd-01h (150 mg) with a yield of 42%.

MS m/z(ESI)：570.1(M+1)。MS m/z(ESI):570.1(M+1).

第八步Step 8

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸的製備Preparation of 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanoic acid

將2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸乙酯Cpd-01h(150 mg, 0.26 mmol)溶於乙醇(5 mL)，再加入3M氫氧化鈉水溶液(5 mL)，室溫下攪拌3小時，反應結束後，調節pH至酸性，室溫下旋蒸除去大部分乙醇，將水溶液凍乾，得到2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸Cpd-01i的粗產品，粗產品直接投下一步。Dissolve 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanoic acid ethyl ester Cpd-01h (150 mg, 0.26 mmol) in ethanol (5 mL), and add 3M aqueous sodium hydroxide solution (5 mL), stirred at room temperature for 3 hours. After the reaction was completed, the pH was adjusted to acidic, most of the ethanol was removed by rotary evaporation at room temperature, and the aqueous solution was freeze-dried to obtain a crude product of 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanoic acid Cpd-01i, which was directly used in the next step.

MS m/z(ESI)：542.1(M+1)。MS m/z(ESI):542.1(M+1).

第九步Step 9

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙醯胺的製備Preparation of 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanamide

向2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙酸Cpd-01i的粗產品中加入DMF(5 mL)，過濾，向濾液中加入HATU(118 mg, 0.31 mmol)，氯化銨(14 mg, 0.31 mmol)以及二異丙基乙基胺(100 mg, 0.78 mmol)，反應液在室溫下攪拌過夜，反應結束後，過濾，經純化得到2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙醯胺Cpd-01(1.15 mg)，收率0.8%。DMF (5 mL) was added to the crude product of 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanoic acid Cpd-01i, filtered, and HATU (118 mg, 0.31 mmol), ammonium chloride (14 mg, 0.31 mmol) and diisopropylethylamine (100 mg, 0.78 mmol), the reaction solution was stirred at room temperature overnight. After the reaction was completed, it was filtered and purified to obtain 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanamide Cpd-01 (1.15 mg) with a yield of 0.8%.

MS m/z(ESI)：541.1(M+1)。MS m/z(ESI):541.1(M+1).

HPLC：100%(214 nm), 99.18%(254 nm)。HPLC: 100% (214 nm), 99.18% (254 nm).

¹H NMR(400 MHz, DMSO- d6)δ 8.94(d, J = 5.1 Hz, 1H), 8.87(s, 1H), 8.62(d, J = 2.1 Hz, 1H), 8.48(s, 1H), 8.35(s, 1H), 8.21(d, J = 5.2 Hz, 1H), 8.14 – 8.07(m, 1H), 6.97(d, J = 24.6 Hz, 2H), 6.85(s, 1H), 5.49(s, 2H), 2.10(s, 3H), 1.97(s, 3H), 1.57(s, 6H). ¹ H NMR (400 MHz, DMSO- d6 )δ 8.94(d, J = 5.1 Hz, 1H), 8.87(s, 1H), 8.62(d, J = 2.1 Hz, 1H), 8.48(s, 1H), 8.35(s, 1H), 8.21(d, J = 5.2 Hz, 1H), 8.14 – 8.07(m, 1H), 6.97(d, J = 24.6 Hz, 2H), 6.85(s, 1H), 5.49(s, 2H), 2.10(s, 3H), 1.97(s, 3H), 1.57(s, 6H).

將上述製備得到的化合物Cpd-01通過掌性拆分的方式獲得化合物Cpd-01A(0.6 mg)。The compound Cpd-01 prepared above was subjected to chiral resolution to obtain compound Cpd-01A (0.6 mg).

MS m/z(ESI)：541.1(M+1)。MS m/z(ESI):541.1(M+1).

HPLC：100%(214 nm), 99.0%(254 nm)。HPLC: 100% (214 nm), 99.0% (254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.89(d, J = 5.0 Hz, 1H), 8.76(s, 1H), 8.42(d, J = 1.9 Hz, 1H), 8.30(d, J = 5.0 Hz, 1H), 8.26(s, 1H), 7.34(td, J = 9.1, 2.1 Hz, 1H), 6.45(s, 2H), 5.44(s, 2H), 2.20(s, 3H), 1.99(s, 3H), 1.75(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.89(d, J = 5.0 Hz, 1H), 8.76(s, 1H), 8.42(d, J = 1.9 Hz, 1H), 8.30(d, J = 5.0 Hz, 1H), 8.26(s, 1H), 7.34(td, J = 9.1, 2.1 Hz, 1H), 6.45(s, 2H), 5.44(s, 2H), 2.20(s, 3H), 1.99(s, 3H), 1.75(s, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK IG_3, 3.0*150mm, 3μm；流動相比例：CO ₂/MeOH(0.1%DEA)= 60/40；柱溫：37度。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALPAK IG_3, 3.0*150mm, 3μm; Mobile phase ratio: CO ₂ /MeOH(0.1%DEA)= 60/40; Column temperature: 37 degrees.

3-胺基-3-亞胺基-2,2-二甲基丙酸乙酯Cpd-01m的製備 Preparation of 3-amino-3-imino-2,2-dimethylpropionic acid ethyl ester Cpd-01m

第一步first step

3-乙氧基-3-亞胺基-2,2-二甲基丙酸乙酯的製備Preparation of ethyl 3-ethoxy-3-imino-2,2-dimethylpropionate

將2-腈基-2-甲基丙酸乙酯Cpd-01j(2 g, 0.014 mol)溶於乙醇(5 mL)，室溫下向反應液中通氯化氫氣體2小時，反應結束後，旋蒸除去乙醇後得到3-乙氧基-3-亞胺基-2,2-二甲基丙酸乙酯Cpd-01k的粗產品(2 g)。2-Nitrile-2-methylpropionic acid ethyl ester Cpd-01j (2 g, 0.014 mol) was dissolved in ethanol (5 mL), and hydrogen chloride gas was passed into the reaction solution at room temperature for 2 hours. After the reaction was completed, ethanol was removed by rotary evaporation to obtain the crude product (2 g) of 3-ethoxy-3-imino-2,2-dimethylpropionic acid ethyl ester Cpd-01k.

MS m/z(ESI)：188.2(M+1)。MS m/z(ESI):188.2(M+1).

第二步Step 2

3-胺基-3-亞胺基-2,2-二甲基丙酸乙酯的製備Preparation of ethyl 3-amino-3-imino-2,2-dimethylpropionate

向3-乙氧基-3-亞胺基-2,2-二甲基丙酸乙酯Cpd-01k的粗品(2 g, 0.011 mol)中加入2M的氨的乙醇溶液(5 mL)，封管攪拌1小時，隨後過濾除去固體，向濾液中加入氨的乙醇溶液(10 mL)，封管攪拌過夜，反應結束後，旋蒸除去乙醇後得到3-胺基-3-亞胺基-2,2-二甲基丙酸乙酯Cpd-01m(1.5 g)。To the crude product (2 g, 0.011 mol) of ethyl 3-ethoxy-3-imino-2,2-dimethylpropionate Cpd-01k, 2M ethanolic ammonia solution (5 mL) was added, the tube was sealed and stirred for 1 hour, and then the solid was removed by filtration. To the filtrate, ethanolic ammonia solution (10 mL) was added, the tube was sealed and stirred overnight. After the reaction was completed, ethanol was removed by rotary evaporation to obtain ethyl 3-amino-3-imino-2,2-dimethylpropionate Cpd-01m (1.5 g).

MS m/z(ESI)：159.2(M+1)。MS m/z (ESI): 159.2 (M+1).

實施例2Embodiment 2

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙腈 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropionitrile

第一步first step

2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙腈的製備Preparation of 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropionitrile

將2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙醯胺Cpd-01(10 mg, 0.018 mmol)溶於二氯甲烷(2 mL)，隨後加入三氟乙酸酐(7.77 mg, 0.037 mmol)以及三乙胺(7.49 mg, 0.074 mmol)，反應液在室溫下攪拌2小時，反應結束後，濃縮經純化得到2-(4-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)嘧啶-2-基)-2-甲基丙腈Cpd-02(2 mg)，收率21%。2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropanamide Cpd-01 (10 mg, 0.018 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of trifluoroacetic anhydride (7.77 mg, 0.037 mmol) and triethylamine (7.49 mg, 0.074 mmol), the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the residue was concentrated and purified to obtain 2-(4-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)pyrimidin-2-yl)-2-methylpropionitrile Cpd-02 (2 mg) with a yield of 21%.

MS m/z(ESI)：523.0(M+1)。MS m/z(ESI):523.0(M+1).

HPLC：99%(214 nm), 99%(254 nm)。HPLC: 99%(214 nm), 99%(254 nm).

¹H NMR(400 MHz, DMSO- d6)δ 9.05(d, J = 5.2 Hz, 1H), 8.90(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.38(s, 1H), 8.34(d, J = 5.2 Hz, 1H), 8.11(td, J = 10.0, 2.3 Hz, 1H), 6.85(s, 1H), 5.49(s, 2H), 2.11(s, 3H), 1.97(s, 3H), 1.80(s, 6H). ¹ H NMR (400 MHz, DMSO- d6 )δ 9.05(d, J = 5.2 Hz, 1H), 8.90(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.38(s, 1H), 8.34(d, J = 5.2 Hz, 1H), 8.11(td, J = 10.0, 2.3 Hz, 1H), 6.85(s, 1H), 5.49(s, 2H), 2.11(s, 3H), 1.97(s, 3H), 1.80(s, 6H).

將上述製備得到的化合物Cpd-02通過掌性拆分的方式獲得化合物Cpd-02A(8mg)和化合物Cpd-02B(6mg)。The compound Cpd-02 prepared above was subjected to chiral separation to obtain compound Cpd-02A (8 mg) and compound Cpd-02B (6 mg).

Cpd-02BCpd-02B

MS m/z(ESI)：523.0(M+1)。MS m/z(ESI):523.0(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

實施例3Embodiment 3

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙腈 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropionitrile

第一步first step

2-(3-溴苯基)-2-甲基丙腈的製備Preparation of 2-(3-bromophenyl)-2-methylpropionitrile

在0℃條件下，向2-(3-溴苯基)乙腈Cpd-17a(1 g, 0.0051 mol)的THF(20 mL)溶液中緩慢加入NaHMDS(2M in THF，2.81 g, 0.0153 mol)。將反應混合物在零度下攪拌30分鐘，然後緩慢加入碘甲烷(1.59 g, 0.01122 mol)。將反應混合物在25℃下攪拌2小時，TLC監測反應結束。然後用水(10 mL)淬滅，並用乙酸乙酯(3×20 mL)萃取。將有機相用無水硫酸鈉乾燥並在減壓下濃縮。將殘餘物用矽膠管柱層析法(流動相：石油醚/乙酸乙酯=10/1)純化後得到2-(3-溴苯基)-2-甲基丙腈Cpd-17b(611 mg)，產率：53.7 %。To a solution of 2-(3-bromophenyl)acetonitrile Cpd-17a (1 g, 0.0051 mol) in THF (20 mL) was added NaHMDS (2M in THF, 2.81 g, 0.0153 mol) slowly at 0°C. The reaction mixture was stirred at zero degrees for 30 minutes, and then iodomethane (1.59 g, 0.01122 mol) was added slowly. The reaction mixture was stirred at 25°C for 2 hours, and the reaction was completed as monitored by TLC. It was then quenched with water (10 mL) and extracted with ethyl acetate (3×20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 2-(3-bromophenyl)-2-methylpropionitrile Cpd-17b (611 mg) with a yield of 53.7%.

¹H NMR(400 MHz, CDCl ₃)δ 7.60(s, 1H), 7.44(dd, J= 13.2, 5.0 Hz, 2H), 7.27(t, J= 7.9 Hz, 1H), 1.72(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 7.60(s, 1H), 7.44(dd, J = 13.2, 5.0 Hz, 2H), 7.27(t, J = 7.9 Hz, 1H), 1.72(s, 6H).

第二步Step 2

2-甲基-2-(3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)苯基)丙腈的製備Preparation of 2-methyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propionitrile

將2-(3-溴苯基)-2-甲基丙腈Cpd-17b (50 mg, 0.2231 mmol), 雙聯頻哪醇硼酸酯(169.96 mg, 0.6693 mmol)溶於1,4-二氧六環(2.5 mL)，隨後加入醋酸鉀(65.69 mg, 0.6693 mmol)和Pd(dppf)Cl ₂(16.32 mg, 0.02231 mmol)。反應混合物在N ₂下90℃反應1小時後，LCMS監測反應結束，反應停止後冷卻至室溫，將反應混合物經矽藻土過濾並濃縮。並用乙酸乙酯(3×10 mL)萃取。將有機相用無水硫酸鈉乾燥並在減壓下濃縮。將殘餘物用矽膠管柱層析法(流動相：石油醚/乙酸乙酯= 10/1)純化後得到2-甲基-2-(3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)苯基)丙腈Cpd-17c(50 mg)，產率：82.4%。 2-(3-Bromophenyl)-2-methylpropionitrile Cpd-17b ( 50 mg, 0.2231 mmol), bis-pinacol boronate (169.96 mg, 0.6693 mmol) were dissolved in 1,4-dioxane (2.5 mL), followed by the addition of potassium acetate (65.69 mg, 0.6693 mmol) and Pd(dppf)Cl ₂ (16.32 mg, 0.02231 mmol). The reaction mixture was reacted at 90°C under N ₂ for 1 hour, and the reaction was completed as monitored by LCMS. After the reaction stopped, the mixture was cooled to room temperature, filtered through diatomaceous earth and concentrated. The mixture was extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give 2-methyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propionitrile Cpd-17c (50 mg) with a yield of 82.4%.

MS m/z(ESI)：272.2(M+1) 。MS m/z(ESI):272.2(M+1).

¹H NMR(400 MHz, CDCl ₃)δ 7.86(s, 1H), 7.76(d, J= 7.3 Hz, 1H), 7.60(d, J= 7.9 Hz, 1H), 7.40(t, J= 7.4 Hz, 1H), 1.75(s, 6H), 1.35(s, 12H). ¹ H NMR (400 MHz, CDCl ₃ )δ 7.86(s, 1H), 7.76(d, J = 7.3 Hz, 1H), 7.60(d, J = 7.9 Hz, 1H), 7.40(t, J = 7.4 Hz, 1H), 1.75(s, 6H), 1.35(s, 12H).

第三步third step

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙腈的製備Preparation of 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropionitrile

向2-甲基-2-(3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)苯基)丙腈Cpd-17c(39.45 mg，0.1455 mmol)和2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮(40 mg，0.097 mmol)的1,4-二氧六環/水(10 mL/2 mL)溶液中加入碳酸鉀(40.22 mg，0.291 mmol)以及Pd(dppf)Cl ₂(7.1 mg, 0.0097 mmol)。反應混合物在N ₂下90℃攪拌5小時，LCMS監測反應結束，反應停止後冷卻至室溫，將反應混合物經矽藻土過濾並濃縮。並用乙酸乙酯(3×10 mL)萃取。將有機相用無水硫酸鈉乾燥並在減壓下濃縮，純化，得2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-5',6-二甲基-2-側氧基-2H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙腈Cpd-17 (20 mg)，產率：39.6 %。 To a solution of 2-methyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propionitrile Cpd-17c (39.45 mg, 0.1455 mmol) and 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one (40 mg, 0.097 mmol) in 1,4-dioxane/water (10 mL/2 mL) was added potassium carbonate (40.22 mg, 0.291 mmol) and Pd(dppf) _Cl2 (7.1 mg, 0.0097 mmol). The reaction mixture was stirred at 90°C under _N2 for 5 hours. The reaction was completed as monitored by LCMS. After the reaction stopped, the mixture was cooled to room temperature and filtered through diatomaceous earth and concentrated. It was extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure and purified to obtain 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropionitrile Cpd-17 ( 20 mg), yield: 39.6%.

MS m/z(ESI)：521.2(M+1)。MS m/z(ESI):521.2(M+1).

HPLC：99.55%(214 nm), 99.38%(254 nm)。HPLC: 99.55%(214 nm), 99.38%(254 nm).

¹H NMR(400 MHz, DMSO)δ 8.75(s, 1H), 8.40(s, 1H), 8.10(s, 1H), 7.94(s, 1H), 7.53(d, J= 24.1 Hz, 3H), 7.33(s, 1H), 6.41(s, 1H), 5.42(s, 2H), 2.18(s, 3H), 2.02(s, 3H), 1.80(s, 6H). ¹ H NMR (400 MHz, DMSO) δ 8.75(s, 1H), 8.40(s, 1H), 8.10(s, 1H), 7.94(s, 1H), 7.53(d, J = 24.1 Hz, 3H), 7.33(s, 1H), 6.41(s, 1H), 5.42(s, 2H), 2.18(s, 3H), 2.02(s, 3H), 1.80(s, 6H).

將上述製備得到的化合物Cpd-17通過掌性拆分的方式獲得化合物Cpd-17A(8mg)和化合物Cpd-17B(6mg)。The compound Cpd-17 prepared above was subjected to chiral separation to obtain compound Cpd-17A (8 mg) and compound Cpd-17B (6 mg).

Cpd-17ACpd-17A

MS m/z(ESI)：521.2(M+1)。MS m/z(ESI):521.2(M+1).

HPLC：100%(214 nm), 99.0%(254 nm)。HPLC: 100% (214 nm), 99.0% (254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.73(s, 1H), 8.40(d, J = 2.2 Hz, 1H), 8.09(s, 1H), 7.92(d, J = 7.6 Hz, 1H), 7.57 – 7.47(m, 3H), 7.36 – 7.30(m, 1H), 6.40(s,1H), 5.42(d, J = 1.5 Hz, 2H), 2.15(s, 3H), 2.00(s, 3H), 1.79(d, J = 3.0 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.73(s, 1H), 8.40(d, J = 2.2 Hz, 1H), 8.09(s, 1H), 7.92(d, J = 7.6 Hz, 1H), 7.57 – 7.47(m, 3H), 7.36 – 7.30(m, 1H), 6.40(s,1H), 5.42(d, J = 1.5 Hz, 2H), 2.15(s, 3H), 2.00(s, 3H), 1.79(d, J = 3.0 Hz, 6H).

實施例4Embodiment 4

2-(3'-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基-2-側氧基吡啶-1(2 H)-基)-4'-甲基-[1,1'-聯苯]-3-基)-2-甲基丙腈 2-(3'-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methyl-2-oxopyridin-1( 2H )-yl)-4'-methyl-[1,1'-biphenyl]-3-yl)-2-methylpropionitrile

第一步first step

N-(5-溴-2-甲基苯基)-2,6-二甲基-4-側氧基-4 H-吡喃-3-甲醯胺的製備 Preparation of N- (5-bromo-2-methylphenyl)-2,6-dimethyl-4-oxo- 4H -pyran-3-carboxamide

向5-溴-2-甲基苯胺Cpd-45a(1 g，5 mmol)的N,N-二甲基苯胺(5 mL)溶液中加入2,2,6-三甲基-4 H-1,3-二英-4-酮(3.84 g，27 mmol)。反應混合物在115℃下攪拌16小時。反應結束後加入水淬滅，用乙酸乙酯萃取(3 × 10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=9/1)純化後得到 N-(5-溴-2-甲基苯基)-2,6-二甲基-4-側氧基-4 H-吡喃-3-甲醯胺Cpd-45b(2 g)，產率：92.59%。 2,2,6-trimethyl- 4H -1,3-dioxin-4-one (3.84 g, 27 mmol) was added to a solution of 5-bromo-2-methylaniline Cpd-45a (1 g, 5 mmol) in N,N-dimethylaniline (5 mL). The reaction mixture was stirred at 115°C for 16 hours. After the reaction was completed, water was added to quench the mixture and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 9/1) to obtain N- (5-bromo-2-methylphenyl)-2,6-dimethyl-4-oxo- 4H -pyran-3-carboxamide Cpd-45b (2 g) with a yield of 92.59%.

MS m/z(ESI)：338.1(M+1)。MS m/z(ESI):338.1(M+1).

第二步Step 2

3-乙醯基-1-(5-溴-2-甲基苯基)-4-羥基-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 3-acetyl-1-(5-bromo-2-methylphenyl)-4-hydroxy-6-methylpyridin-2( 1H )-one

將化合物Cpd-45b(2 g，10 mmol)溶解在水(15 mL)和鹽酸/1,4-二氧六環(35 mL)中。反應混合物在85℃下攪拌16小時。反應結束後加入水淬滅，用乙酸乙酯萃取(3×10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=9/1)純化後得到3-乙醯基-1-(5-溴-2-甲基苯基)-4-羥基-6-甲基吡啶-2(1 H)-酮Cpd-45c(880 mg)，產率：23.76%。 Compound Cpd-45b (2 g, 10 mmol) was dissolved in water (15 mL) and hydrochloric acid/1,4-dioxane (35 mL). The reaction mixture was stirred at 85°C for 16 hours. After the reaction was completed, water was added to quench and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 9/1) to obtain 3-acetyl-1-(5-bromo-2-methylphenyl)-4-hydroxy-6-methylpyridin-2(1 H )-one Cpd-45c (880 mg) with a yield of 23.76%.

MS m/z(ESI)：337.0(M+1)。MS m/z(ESI):337.0(M+1).

第三步third step

1-(5-溴-2-甲基苯基)-4-羥基-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-bromo-2-methylphenyl)-4-hydroxy-6-methylpyridin-2( 1H )-one

將化合物Cpd-45c(880 mg，2 mmol)溶解在濃硫酸(12 mL)中。反應混合物在100℃下攪拌3小時。反應結束後加入飽和碳酸氫鈉溶液(10 mL)淬滅，用乙酸乙酯萃取(3 × 10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)純化後得到1-(5-溴-2-甲基苯基)-4-羥基-6-甲基吡啶-2(1 H)-酮Cpd-45d(360 mg)，產率：42.08%。 Compound Cpd-45c (880 mg, 2 mmol) was dissolved in concentrated sulfuric acid (12 mL). The reaction mixture was stirred at 100 °C for 3 hours. After the reaction was completed, saturated sodium bicarbonate solution (10 mL) was added to quench the reaction and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 1-(5-bromo-2-methylphenyl)-4-hydroxy-6-methylpyridin-2(1 H )-one Cpd-45d (360 mg) with a yield of 42.08%.

MS m/z(ESI)：295.0(M+1)。MS m/z (ESI): 295.0 (M+1).

第四步the fourth step

1-(5-溴-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-bromo-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methylpyridin-2( 1H )-one

將化合物Cpd-45d(360 mg，1 mmol)、2-(溴甲基)-3,5-二氟吡啶(254 mg，1 mmol)和碳酸鉀(169 mg，1 mmol)溶解在N,N-二甲基甲醯胺(12 mL)中，在60℃攪拌16小時。反應結束後加入水淬滅，用乙酸乙酯萃取(3 × 10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=5/1)純化後得到1-(5-溴-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基吡啶-2(1 H)-酮Cpd-45e(500 mg)，產率：87.29%。 Compound Cpd-45d (360 mg, 1 mmol), 2-(bromomethyl)-3,5-difluoropyridine (254 mg, 1 mmol) and potassium carbonate (169 mg, 1 mmol) were dissolved in N,N-dimethylformamide (12 mL) and stirred at 60°C for 16 hours. After the reaction was completed, water was added to quench and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain 1-(5-bromo-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methylpyridin-2( 1H )-one Cpd-45e (500 mg) in a yield of 87.29%.

MS m/z(ESI)：421.0(M+1)。MS m/z(ESI):421.0(M+1).

第五步the fifth step

1-(5-溴-2-甲基苯基)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-bromo-2-methylphenyl)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methylpyridin-2( 1H )-one

將化合物Cpd-45e(500 mg，1.1 mmol)、N-氯代丁二醯亞胺(174 mg，1.3 mmol)和二氯乙酸(15 mg，0.1 mmol)溶解在異丙醇(12 mL)中，在60℃攪拌3小時。反應結束後，用水淬滅反應，乙酸乙酯(3×10 mL)萃取。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)純化後得到1-(5-溴-2-甲基苯基)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基吡啶-2(1 H)-酮Cpd-45f(350 mg)，產率：58.24%。 Compound Cpd-45e (500 mg, 1.1 mmol), N-chlorosuccinimide (174 mg, 1.3 mmol) and dichloroacetic acid (15 mg, 0.1 mmol) were dissolved in isopropanol (12 mL) and stirred at 60 °C for 3 hours. After the reaction was completed, the reaction was quenched with water and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 1-(5-bromo-2-methylphenyl)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methylpyridin-2( 1H )-one Cpd-45f (350 mg) in a yield of 58.24%.

MS m/z(ESI)：457.0(M+1)。MS m/z (ESI): 457.0 (M+1).

第六步Step 6

2-(3'-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基-2-側氧基吡啶-1(2 H)-基)-4'-甲基-[1,1'-聯苯]-3-基)-2-甲基丙腈的製備 Preparation of 2-(3'-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methyl-2-oxopyridin-1( 2H )-yl)-4'-methyl-[1,1'-biphenyl]-3-yl)-2-methylpropionitrile

將化合物Cpd-45f(50 mg，0.1 mmol)溶解在四氫呋喃：水=2：1(9 mL)中，加入化合物Cpd-17c(75 mg，0.1 mmol)，碳酸鈉(39 mg， 0.3 mmol)和四(三苯基膦)鈀(21 mg，0.01mmol)。將反應混合物在90℃下攪拌1小時。反應結束後，用水淬滅反應，用乙酸乙酯(3×10 mL)萃取。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮。濃縮液通過製備純化並凍乾得到2-(3’-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-6-甲基-2-側氧基吡啶-1(2 H)-基)-4’-甲基-[1,1’-聯苯]-3-基)-2-甲基丙腈 (化合物Cpd-45 )(2.21 mg)，產率：2.28%。 Compound Cpd-45f (50 mg, 0.1 mmol) was dissolved in tetrahydrofuran: water = 2:1 (9 mL), and compound Cpd-17c (75 mg, 0.1 mmol), sodium carbonate (39 mg, 0.3 mmol) and tetrakis(triphenylphosphine)palladium (21 mg, 0.01 mmol) were added. The reaction mixture was stirred at 90 °C for 1 hour. After the reaction was completed, the reaction was quenched with water and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was purified by preparative purification and freeze-dried to give 2-(3'-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-6-methyl-2-oxopyridin-1( 2H )-yl)-4'-methyl-[1,1'-biphenyl]-3-yl)-2-methylpropionitrile ( Compound Cpd-45 ) (2.21 mg) in a yield of 2.28%.

MS m/z(ESI)：520.1(M+1)。MS m/z(ESI):520.1(M+1).

¹H NMR(400 MHz, DMSO- d6)δ 8.40(d, J= 2.2 Hz, 1H), 7.64(s, 1H), 7.60(dd, J= 7.9, 1.6 Hz, 1H), 7.53 – 7.50(m, 1H), 7.47 – 7.43(m, 3H), 7.35 – 7.28(m, 2H), 6.35(s, 1H), 5.40(d, J= 1.4 Hz, 2H), 2.12(s, 3H), 1.99(s, 3H), 1.77(s, 6H). ¹ H NMR (400 MHz, DMSO- d6 )δ 8.40(d, J = 2.2 Hz, 1H), 7.64(s, 1H), 7.60(dd, J = 7.9, 1.6 Hz, 1H), 7.53 – 7.50(m, 1H), 7.47 – 7.43(m, 3H), 7.35 – 7.28(m, 2H), 6.35(s, 1H), 5.40(d, J = 1.4 Hz, 2H), 2.12(s, 3H), 1.99(s, 3H), 1.77(s, 6H).

實施例5Embodiment 5

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 3-Chloro-4-((3,5-difluoropyridin-2-yl)methoxy _- d2 )-3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

第一步first step

2-氯-3-氟-5-甲基吡啶-4-胺的製備Preparation of 2-chloro-3-fluoro-5-methylpyridin-4-amine

向2-氯-3-氟-5-碘吡啶-4-胺Cpd-40a(50 g, 0.18 mol)，甲基硼酸(13.2 g, 0.22 mol)，Pd(dppf)Cl ₂(13.4 g, 0.018 mol)以及碳酸銫(119.6 g, 0.36 mol)中加入1,4-二氧六環(500 mL)，反應液在100攝氏度下攪拌16小時，反應結束後，過濾，濃縮後經矽膠管柱(石油醚/乙酸乙酯=3/1)分離純化得到2-氯-3-氟-5-甲基吡啶-4-胺Cpd-40b(17.4 g)，收率60%。 1,4-Dioxane (500 mL) was added to 2-chloro-3-fluoro-5-iodopyridin-4-amine Cpd-40a (50 g, 0.18 mol), methylboric acid (13.2 g, 0.22 mol), Pd(dppf)Cl ₂ (13.4 g, 0.018 mol) and cesium carbonate (119.6 g, 0.36 mol). The reaction solution was stirred at 100 degrees Celsius for 16 hours. After the reaction was completed, the product was filtered, concentrated and purified by silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 2-chloro-3-fluoro-5-methylpyridin-4-amine Cpd-40b (17.4 g) with a yield of 60%.

MS m/z(ESI)：161.1(M+1)。MS m/z(ESI):161.1(M+1).

第二步Step 2

N-(2-氯-3-氟-5-甲基吡啶-4-基)-2,6-二甲基-4-側氧基-4 H-吡喃-3-甲醯胺的製備 Preparation of N- (2-chloro-3-fluoro-5-methylpyridin-4-yl)-2,6-dimethyl-4-oxo- 4H -pyran-3-carboxamide

向2-氯-3-氟-5-甲基吡啶-4-胺Cpd-40b(8.7 g, 0.054 mol)以及2,2,6-三甲基-4 H-1,3-二噁英-4-酮(38.5 g, 0.27 mol)中加入DMA(30 mL)，反應液在115攝氏度下攪拌16小時，反應結束後，冷卻至室溫加入水(50 ml)，攪拌，將反應液抽濾，濾渣為 N-(2-氯-3-氟-5-甲基吡啶-4-基)-2,6-二甲基-4-側氧基-4 H-吡喃-3-甲醯胺Cpd-40c，粗品直接投下一步。 DMA (30 mL) was added to 2-chloro-3-fluoro-5-methylpyridin-4-amine Cpd-40b (8.7 g, 0.054 mol) and 2,2,6-trimethyl- 4H -1,3-dioxin-4-one (38.5 g, 0.27 mol). The reaction solution was stirred at 115°C for 16 hours. After the reaction was completed, it was cooled to room temperature and water (50 ml) was added. The mixture was stirred and filtered. The residue was N- (2-chloro-3-fluoro-5-methylpyridin-4-yl)-2,6-dimethyl-4-oxo- 4H -pyran-3-carboxamide Cpd-40c. The crude product was directly used in the next step.

MS m/z(ESI)：311.1(M+1)。MS m/z(ESI):311.1(M+1).

第三步third step

3-乙醯基-2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向上一步的粗品中加入水(50 mL)，加入4 M 鹽酸的二氧六環溶液(50 mL)，反應液在85攝氏度下攪拌16小時，反應結束後，旋蒸除去部分二氧六環，抽濾，濾渣為3-乙醯基-2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-40d(11.5 g)。 Water (50 mL) was added to the crude product of the previous step, and a 4 M hydrochloric acid solution of dioxane (50 mL) was added. The reaction solution was stirred at 85 °C for 16 hours. After the reaction was completed, some dioxane was removed by rotary evaporation and filtered. The residue was 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-40d (11.5 g).

MS m/z(ESI)：311.1(M+1)。MS m/z(ESI):311.1(M+1).

第四步the fourth step

2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向3-乙醯基-2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-40d(11.5 g, 0.037 mol)中加入濃硫酸(50 mL)，反應液在100攝氏度下攪拌16小時，反應結束後，冷卻至室溫，反應液加入冰水中，用NaOH調pH至9-10，再用濃鹽酸調pH至3-4，抽濾，所得濾渣為2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-40e(13 g)。 Concentrated sulfuric acid (50 mL) was added to 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-40d (11.5 g, 0.037 mol), and the reaction solution was stirred at 100 degrees Celsius for 16 hours. After the reaction was completed, it was cooled to room temperature, and the reaction solution was added to ice water. The pH was adjusted to 9-10 with NaOH, and then to 3-4 with concentrated hydrochloric acid. The solution was filtered to obtain 2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-40e (13 g).

MS m/z(ESI)：269.0(M+1)。MS m/z (ESI): 269.0 (M+1).

第五步the fifth step

3,5-二氟吡啶-2-甲酸乙酯的製備Preparation of ethyl 3,5-difluoropyridine-2-carboxylate

將3,5-二氟吡啶-2-甲酸Cpd-40f(10 g, 62.9 mmol)溶於二氯甲烷(50 mL)，在0度下滴加二氯亞碸(15 g, 125.8 mmol)，滴加完成後，於50度下反應5小時，反應結束後，加熱濃縮，隨後加入乙醇(50 mL)，在室溫條件下攪拌1小時。反應結束後，濃縮，粗產品用矽膠管柱(石油醚)純化得到3,5-二氟吡啶-2-甲酸乙酯 Cpd-40g(10 g)，產率：81%。3,5-Difluoropyridine-2-carboxylic acid Cpd-40f (10 g, 62.9 mmol) was dissolved in dichloromethane (50 mL), and dichlorosulfonyl chloride (15 g, 125.8 mmol) was added dropwise at 0 degrees. After the addition was completed, the reaction was carried out at 50 degrees for 5 hours. After the reaction was completed, the mixture was heated and concentrated, and then ethanol (50 mL) was added and stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated and the crude product was purified with a silica gel column (petroleum ether) to obtain 3,5-difluoropyridine-2-carboxylic acid ethyl ester Cpd-40g (10 g), with a yield of 81%.

MS m/z(ESI)：188.1(M+1)。MS m/z(ESI):188.1(M+1).

第六步Step 6

(3,5-二氟吡啶-2-基)甲- d ₂-醇的製備 Preparation of (3,5-difluoropyridin-2-yl)methan- d ₂ -ol

將3,5-二氟吡啶-2-甲酸乙酯 Cpd-40g(500 mg，2.67 mmol)溶於氘代甲醇(5 mL)和四氫呋喃(10 mL)，在0度下加入硼氘化鈉(145 mg, 3.47 mmol)，在25度下反應2小時。反應結束後，用重水(10 mL)淬滅反應，用乙酸乙酯(3 × 20 mL)萃取，合併有機相，用飽和食鹽水(20 mL)洗，有機相用無水硫酸鈉乾燥，過濾，濃縮得到粗品(3,5-二氟吡啶-2-基)甲- d ₂-醇 Cpd-40h(300 mg)，產率：68%。 3,5-Difluoropyridine-2-carboxylic acid ethyl ester Cpd-40g (500 mg, 2.67 mmol) was dissolved in deuterated methanol (5 mL) and tetrahydrofuran (10 mL), sodium borodeuteride (145 mg, 3.47 mmol) was added at 0 degrees, and the mixture was reacted at 25 degrees for 2 hours. After the reaction was completed, the reaction was quenched with heavy water (10 mL), extracted with ethyl acetate (3 × 20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude (3,5-difluoropyridin-2-yl)methan- d2 _- ol Cpd-40h (300 mg), with a yield of 68%.

MS m/z(ESI)：148.1(M+1)。MS m/z(ESI):148.1(M+1).

¹H NMR(400 MHz, DMSO) δ8.46(d, J= 2.4 Hz, 1H), 7.96 – 7.81(m, 1H), 5.34(s, 1H). ¹ H NMR (400 MHz, DMSO) δ 8.46 (d, J = 2.4 Hz, 1H), 7.96 – 7.81 (m, 1H), 5.34 (s, 1H).

第七步Step 7

2-(氯甲基- d ₂)-3,5-二氟吡啶的製備 Preparation of 2-(chloromethyl- d ₂ )-3,5-difluoropyridine

將(3,5-二氟吡啶-2-基)甲- d ₂-醇 Cpd-40h(200 mg, 1.359 mmol)和N,N-二甲基甲醯胺(10 mg, 0.136 mmol)溶於二氯甲烷(10 mL)，在0度下滴加二氯亞碸(485 mg, 4.078 mmol)，在25度下反應2小時。反應結束後，反應液濃縮得到2-(氯甲基- d ₂)-3,5-二氟吡啶 Cpd-40i(200 mg)，產率：87%。 (3,5-Difluoropyridin-2-yl)methan- d ₂ -ol Cpd-40h (200 mg, 1.359 mmol) and N,N-dimethylformamide (10 mg, 0.136 mmol) were dissolved in dichloromethane (10 mL), and dichlorosulfonyl chloride (485 mg, 4.078 mmol) was added dropwise at 0 degrees, and the mixture was reacted at 25 degrees for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain 2-(chloromethyl- d ₂ )-3,5-difluoropyridine Cpd-40i (200 mg), with a yield of 87%.

¹H NMR(400 MHz, DMSO) δ8.54(d, J= 2.4 Hz, 1H), 8.07-8.02(m, 1H). ¹ H NMR (400 MHz, DMSO) δ 8.54 (d, J = 2.4 Hz, 1H), 8.07-8.02 (m, 1H).

第八步Step 8

2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- d ₂ )-3'-fluoro-5',6-dimethyl-2 H -[1,4'-bipyridinyl]-2-one

將2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-40e(200 mg, 0.744 mmol)、2-(氯甲基- d ₂)-3,5-二氟吡啶 Cpd-40i(135 mg, 0.819 mmol)、18-冠醚-6(39 mg, 0.149 mmol)和碳酸鉀(206 mg, 1.489 mmol)溶於N,N-二甲基甲醯胺(10 mL)，反應在60度下攪拌3小時。反應結束後，用水(20 mL)洗，用乙酸乙酯(3 × 20 mL)萃取，合併有機相，用飽和食鹽水(20 mL)洗，有機相用無水硫酸鈉乾燥，過濾，濃縮。粗產品用矽膠管柱(乙酸乙酯/石油醚=85%)純化得到2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-40j(200 mg)，產率：69%。 2'-Chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd- _40e (200 mg, 0.744 mmol), 2-(chloromethyl- d2 )-3,5-difluoropyridine Cpd-40i (135 mg, 0.819 mmol), 18-crown-6 (39 mg, 0.149 mmol) and potassium carbonate (206 mg, 1.489 mmol) were dissolved in N,N-dimethylformamide (10 mL) and the reaction was stirred at 60 degrees for 3 hours. After the reaction was completed, the mixture was washed with water (20 mL), extracted with ethyl acetate (3 × 20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 85%) to obtain 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- d2 )-3'-fluoro- ₅ ',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-40j (200 mg), yield: 69%.

MS m/z(ESI)：398.1(M+1)。MS m/z (ESI): 398.1 (M+1).

第九步Step 9

2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)- 3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy _- d2 )-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-40j(180 mg, 0.452 mmol)和N-氯代丁二醯亞胺(72 mg, 0.543 mmol)溶於N,N-二甲基甲醯胺(10 mL)，在60度下反應5小時。反應結束後，反應結束後，用硫代硫酸鈉水溶液(20 mL)淬滅，用乙酸乙酯(3 × 20 mL)萃取，合併有機相，用飽和食鹽水(20 mL)洗，有機相用無水硫酸鈉乾燥，過濾，濃縮。粗產品用矽膠管柱(乙酸乙酯/石油醚=85%)純化得到2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-40k(150 mg)，產率：75%。 2'-Chloro-4-((3,5-difluoropyridin-2-yl)methoxy _- d2 )-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-40j (180 mg, 0.452 mmol) and N-chlorobutanediamide (72 mg, 0.543 mmol) were dissolved in N,N-dimethylformamide (10 mL) and reacted at 60 degrees for 5 hours. After the reaction was completed, it was quenched with sodium thiosulfate aqueous solution (20 mL), extracted with ethyl acetate (3 × 20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 85%) to give 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy- d2 )-3'-fluoro- ₅ ',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-40k (150 mg) in a yield of 75%.

MS m/z(ESI)：432.0(M+1)。MS m/z(ESI):432.0(M+1).

第十步Step 10

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy _- d2 )-3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-40k (1.7 g, 5.8 mmol)、(3-(2-羥基丙烷-2-基)苯基)硼酸Cpd-40m(90 mg, 0.501 mmol)、四(三苯基磷)鈀(48 mg, 0.042 mmol)和碳酸鈉(44 mg, 0.834 mmol)溶於四氫呋喃：水 = 3：1(12 mL)，在90度下攪拌2小時。反應結束後，過濾，濃縮。粗產品通過製備HPLC純化(FA，流動相：ACN：H ₂O(0.1% FA)=49%：51%)，得到3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- d ₂)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)- 5',6-二甲基2 H-[1,4'-聯吡啶]-2-酮 Cpd-40(20.86 mg)。產率：3.85%。 2',3-Dichloro-4-((3,5-difluoropyridin-2-yl)methoxy _- d2 )-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-40k ( 1.7 g, 5.8 mmol), (3-(2-hydroxypropane-2-yl)phenyl)boronic acid Cpd-40m (90 mg, 0.501 mmol), tetrakis(triphenylphosphine)palladium (48 mg, 0.042 mmol) and sodium carbonate (44 mg, 0.834 mmol) were dissolved in tetrahydrofuran: water = 3:1 (12 mL) and stirred at 90 degrees for 2 hours. After the reaction was completed, the mixture was filtered and concentrated. The crude product was purified by preparative HPLC (FA, mobile phase: ACN: _H2O (0.1% FA) = 49%: 51%) to give 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- d2 ₎ -3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6- dimethyl2H- [1,4'-bipyridinyl]-2-one Cpd-40 (20.86 mg). Yield: 3.85%.

Cpd-40Cpd-40

MS m/z(ESI)：532.1(M+1)。MS m/z(ESI):532.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

1H NMR(400 MHz, CDCl ₃) δ8.58(s, 1H), 8.42(d, J= 2.4 Hz, 1H), 8.10(d, J= 1.6 Hz, 1H), 7.83(dd, J= 7.6, 1.0 Hz, 1H), 7.60(ddd, J= 7.6, 1.8, 1.0 Hz, 1H), 7.47(t, J= 7.6 Hz, 1H), 7.34(ddd, J= 9.2, 8.0, 2.4 Hz, 1H), 6.44(d, J= 0.8 Hz, 1H), 2.19(s, 3H), 2.05(s, 3H), 2.02(s, 1H), 1.64(s, 6H). 1H NMR (400 MHz, CDCl ₃ ) δ 8.58(s, 1H), 8.42(d, J = 2.4 Hz, 1H), 8.10(d, J = 1.6 Hz, 1H), 7.83(dd, J = 7.6, 1.0 Hz, 1H), 7.60(ddd, J = 7.6, 1.8, 1.0 Hz, 1H), 7.47(t, J = 7.6 Hz, 1H), 7.34(ddd, J = 9.2, 8.0, 2.4 Hz, 1H), 6.44(d, J = 0.8 Hz, 1H), 2.19(s, 3H), 2.05(s, 3H), 2.02(s, 1H), 1.64(s, 6H).

化合物Cpd-40經掌性拆分(流動相：CO ₂/MeOH(0.1%DEA)=70/30)以及製備得到Cpd-40A(t _R= 0.940 min)(4 mg)；以及Cpd-40B(t _R= 1.513 min)(6 mg)。 Compound Cpd-40 was subjected to chiral separation (mobile phase: CO ₂ /MeOH (0.1% DEA) = 70/30) to prepare Cpd-40A (t _R = 0.940 min) (4 mg); and Cpd-40B (t _R = 1.513 min) (6 mg).

Cpd-40ACpd-40A

MS m/z(ESI)：532.1(M+1)。MS m/z(ESI):532.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.58(s, 1H), 8.41(d, J= 2.4 Hz, 1H), 8.10(d, J= 1.2 Hz, 1H), 7.85 – 7.78(m, 1H), 7.60(d, J= 8.0 Hz, 1H), 7.47(t, J= 7.8 Hz, 1H), 7.38 – 7.29(m, 1H), 6.44(s, 1H), 2.19(s, 3H), 2.04(s, 3H), 1.64(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.58(s, 1H), 8.41(d, J = 2.4 Hz, 1H), 8.10(d, J = 1.2 Hz, 1H), 7.85 – 7.78(m, 1H), 7.60(d, J = 8.0 Hz, 1H), 7.47(t, J = 7.8 Hz, 1H), 7.38 – 7.29(m, 1H), 6.44(s, 1H), 2.19(s, 3H), 2.04(s, 3H), 1.64(s, 6H).

Cpd-40BCpd-40B

MS m/z(ESI)：532.2(M+1)。MS m/z(ESI):532.2(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.59(s, 1H), 8.39(d, J= 2.4 Hz, 1H), 8.11(s, 1H), 7.80(d, J= 7.0 Hz, 1H), 7.59(d, J= 8.0 Hz, 1H), 7.46(t, J= 7.8 Hz, 1H), 7.31(ddd, J= 9.0, 8.0, 2.4 Hz, 1H), 6.43(s, 1H), 2.19(s, 3H), 2.03(s, 3H), 1.62(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.59(s, 1H), 8.39(d, J = 2.4 Hz, 1H), 8.11(s, 1H), 7.80(d, J = 7.0 Hz, 1H), 7.59(d, J = 8.0 Hz, 1H), 7.46(t, J = 7.8 Hz, 1H), 7.31(ddd, J = 9.0, 8.0, 2.4 Hz, 1H), 6.43(s, 1H), 2.19(s, 3H), 2.03(s, 3H), 1.62(s, 6H).

掌性拆分條件：儀器品牌：Waters Acquity UPCC 製備級管柱型號：Daicel CHIRALPAK AD_3, 3*150mm, 3μm 流動相比例：A/B：CO ₂/MeOH(0.1%DEA)=70/30 流速比：2.0 ml/min 柱溫：37度 Chiral separation conditions: Instrument brand: Waters Acquity UPCC Preparation grade column model: Daicel CHIRALPAK AD_3, 3*150mm, 3μm Mobile phase ratio: A/B: CO ₂ /MeOH(0.1%DEA)=70/30 Flow rate ratio: 2.0 ml/min Column temperature: 37 degrees

實施例6Embodiment 6

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(3-(2-羥基丙-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 3-Chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

第一步first step

2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-40e(4.5 g, 0.017 mol)、2-(溴甲基)-3,5-二氟吡啶(3.9 g, 0.019 mol)、碳酸鉀(4.7 g, 0.034 mol)以及18-冠醚-6(4.5 g, 0.017 mol)中加入DMF(30 mL)，反應液在60攝氏度下攪拌16小時，反應結束後，抽濾，濾液中加入溴化鋰溶液，乙酸乙酯萃取，飽和食鹽水洗，有機相乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53a(3.1 g)，收率46%。 DMF (30 mL) was added to 2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-40e (4.5 g, 0.017 mol), 2-(bromomethyl)-3,5-difluoropyridine (3.9 g, 0.019 mol), potassium carbonate (4.7 g, 0.034 mol) and 18-crown-6 (4.5 g, 0.017 mol). mL), the reaction solution was stirred at 60 degrees Celsius for 16 hours. After the reaction was completed, it was filtered, lithium bromide solution was added to the filtrate, extracted with ethyl acetate, washed with saturated brine, and the organic phase was dried and concentrated and then separated and purified by silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53a (3.1 g) with a yield of 46%.

MS m/z(ESI)：396.0(M+1)。MS m/z (ESI): 396.0 (M+1).

第二步Step 2

2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53a(3.1 g, 7.8 mmol)以及N-氯代丁二醯亞胺(NCS)(1.15 g, 8.6 mmol)中加入DMF(15 mL)，再滴加數滴二氯乙酸，反應液在60攝氏度下攪拌1小時，反應結束後，加入硫代硫酸鈉水溶液，溴化鋰水溶液洗滌，有機相再用飽和氯化鈉洗，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53b(2.8 g)，收率83%。 DMF (15 μL) was added to 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53a (3.1 g, 7.8 mmol) and N-chlorosuccinimide (NCS) (1.15 g, 8.6 mmol). mL), and then a few drops of dichloroacetic acid were added dropwise. The reaction solution was stirred at 60 degrees Celsius for 1 hour. After the reaction was completed, sodium thiosulfate aqueous solution was added, and lithium bromide aqueous solution was washed. The organic phase was then washed with saturated sodium chloride, dried, concentrated, and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53b (2.8 g) with a yield of 83%.

MS m/z(ESI)：430.0(M+1)。MS m/z(ESI):430.0(M+1).

第三步third step

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(3-(2-羥基丙-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53b(240 mg, 0.56 mmol)、(3-(2-羥基丙-2-基)苯基)硼酸Cpd-40m(120 mg, 0.67 mmol)、四三苯基膦鈀(65 mg, 0.056 mmol)以及碳酸鈉(119 mg, 1.12 mmol)中加入四氫呋喃/水=2/1(6 mL)，反應液在90攝氏度下攪拌1小時，反應結束後，乙酸乙酯萃取，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到粗產品，再經製備HPLC純化(FA，流動相：ACN：H ₂O(0.1% FA)=49%：51%)得到3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(3-(2-羥基丙-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53(135 mg)，收率45%。 To 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53b (240 mg, 0.56 mmol), (3-(2-hydroxypropan-2-yl)phenyl)boronic acid Cpd-40m (120 mg, 0.67 mmol), tetrakistriphenylphosphine palladium (65 mg, 0.056 mmol) and sodium carbonate (119 mg, 1.12 mmol) was added tetrahydrofuran/water = 2/1 (6 The reaction solution was stirred at 90 degrees Celsius for 1 hour. After the reaction was completed, it was extracted with ethyl acetate, dried, concentrated, and separated and purified by silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain a crude product, which was then purified by preparative HPLC (FA, mobile phase: ACN: _H2O (0.1% FA) = 49%: 51%) to obtain 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(3-(2-hydroxypropyl-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53 (135 mg) in a yield of 45%.

MS m/z(ESI)：530.1(M+1)。MS m/z (ESI): 530.1 (M+1).

HPLC：99.52%(214 nm), 99.72%(254 nm)。HPLC: 99.52%(214 nm), 99.72%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.57(s, 1H), 8.40(d, J= 2.1 Hz, 1H), 8.08(s, 1H), 7.81(d, J= 7.4 Hz, 1H), 7.59(d, J= 7.8 Hz, 1H), 7.46(t, J= 7.6 Hz, 1H), 7.36 – 7.29(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.19(s, 3H), 2.04(s, 3H), 2.01(s, 1H), 1.63(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.57(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 8.08(s, 1H), 7.81(d, J = 7.4 Hz, 1H), 7.59(d, J = 7.8 Hz, 1H), 7.46(t, J = 7.6 Hz, 1H), 7.36 – 7.29(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.19(s, 3H), 2.04(s, 3H), 2.01(s, 1H), 1.63(s, 6H).

化合物Cpd-53經掌性拆分得到Cpd-53A(60.49 mg, t _R= 1.535 min)以及Cpd-53B(51.26 mg, t _R= 1.950 min)。 Compound Cpd-53 was subjected to chiral separation to obtain Cpd-53A (60.49 mg, t _R = 1.535 min) and Cpd-53B (51.26 mg, t _R = 1.950 min).

Cpd-53ACpd-53A

MS m/z(ESI)：530.1(M+1)。MS m/z (ESI): 530.1 (M+1).

HPLC：99.46%(214 nm), 99.56%(254 nm)。HPLC: 99.46%(214 nm), 99.56%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.55(s, 1H), 8.40(d, J= 2.1 Hz, 1H), 8.07(s, 1H), 7.81(d, J= 7.5 Hz, 1H), 7.58(d, J= 7.9 Hz, 1H), 7.45(t, J= 7.8 Hz, 1H), 7.37 – 7.28(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.18(s, 3H), 2.03(s, 3H), 1.63(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.55(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 8.07(s, 1H), 7.81(d, J = 7.5 Hz, 1H), 7.58(d, J = 7.9 Hz, 1H), 7.45(t, J = 7.8 Hz, 1H), 7.37 – 7.28(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.18(s, 3H), 2.03(s, 3H), 1.63(s, 6H).

Cpd-53BCpd-53B

MS m/z(ESI)：530.1(M+1)。MS m/z (ESI): 530.1 (M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.55(s, 1H), 8.40(d, J= 2.1 Hz, 1H), 8.07(s, 1H), 7.81(d, J= 7.6 Hz, 1H), 7.58(d, J= 7.8 Hz, 1H), 7.45(t, J= 7.8 Hz, 1H), 7.36 – 7.28(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.17(s, 3H), 2.03(s, 3H), 1.62(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.55(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 8.07(s, 1H), 7.81(d, J = 7.6 Hz, 1H), 7.58(d, J = 7.8 Hz, 1H), 7.45(t, J = 7.8 Hz, 1H), 7.36 – 7.28(m, 1H), 6.42(s, 1H), 5.41(s, 2H), 2.17(s, 3H), 2.03(s, 3H), 1.62(s, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號： Daicel CHIRALCEL OX, 250mm 30 mm I.D., 10μm；流動相：CO ₂/MeOH[0.2%NH ₃(7M Solution in MeOH)]= 75/25；流速比：2.0 ml/min；柱溫：37度。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALCEL OX, 250mm 30 mm ID, 10μm; Mobile phase: CO ₂ /MeOH [0.2% NH ₃ (7M Solution in MeOH)] = 75/25; Flow rate ratio: 2.0 ml/min; Column temperature: 37 degrees.

實施例7Embodiment 7

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)- N,2-二甲基丙醯胺 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)- N ,2-dimethylpropionamide

第一步first step

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸乙酯的製備 Preparation of ethyl 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoate

向2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53b(500 mg, 1.2 mmol)、2-甲基-2-(3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)苯基)丙酸乙酯(445 mg, 1.4 mmol)、四三苯基膦鈀(139 mg, 0.12 mmol)以及碳酸鈉(254 mg, 2.4 mmol)中加入四氫呋喃/水=1/1(4 mL)，反應液在90攝氏度下攪拌2小時，反應結束後，乙酸乙酯萃取，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/3)分離純化得到2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸乙酯Cpd-54a(253 mg)，收率36%。 To 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53b (500 mg, 1.2 mmol), ethyl 2-methyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate (445 mg, 1.4 mmol), palladium tetrakistriphenylphosphine (139 mg, 0.12 mmol) and sodium carbonate (254 mg, 2.4 mmol) was added tetrahydrofuran/water = 1/1 (4 The reaction solution was stirred at 90 °C for 2 h. After the reaction was completed, the product was extracted with ethyl acetate, dried, concentrated, and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/3) to obtain 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid ethyl ester Cpd-54a (253 mg) in a yield of 36%.

MS m/z(ESI)：586.2(M+1)。MS m/z(ESI):586.2(M+1).

第二步Step 2

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸的製備 Preparation of 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid

將2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸乙酯Cpd-54a(270 mg, 0.46 mmol)溶解在10 N HCl(30 mL)中，反應液在50攝氏度下攪拌6小時，反應結束後，使用氫氧化鈉調節反應液pH=10左右，乙酸乙酯萃取，合併水相，再用鹽酸調節水相pH=3左右，乙酸乙酯萃取，合併有機相，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/3)分離純化得到2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸Cpd-54b (196 mg)，收率76%。 Ethyl 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoate Cpd-54a (270 mg, 0.46 mmol) was dissolved in 10 N HCl (30 The reaction solution was stirred at 50 degrees Celsius for 6 hours. After the reaction was completed, sodium hydroxide was used to adjust the pH of the reaction solution to about 10, and ethyl acetate was used for extraction. The aqueous phases were combined, and then hydrochloric acid was used to adjust the pH of the aqueous phase to about 3. Ethyl acetate was used for extraction. The organic phases were combined, dried and concentrated, and then separated and purified by silica gel column (petroleum ether/ethyl acetate = 1/3) to obtain 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid Cpd-54b ( 196 mg), with a yield of 76%.

MS m/z(ESI)：558.1(M+1)。MS m/z(ESI):558.1(M+1).

第三步third step

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)- N,2-二甲基丙醯胺的製備 Preparation of 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl) -N ,2-dimethylpropionamide

向2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸Cpd-54b (200 mg, 0.36 mmol)、甲胺鹽酸鹽(33 mg, 0.4 mmol)、2-(7-氮雜苯並三唑-1-基)- N, N, N', N'-四甲基脲六氟磷酸鹽(140 mg, 0.36 mmol)、三乙胺(0.1 mL, 0.72 mmol)中加入DMF(2 mL)，反應液在室溫攪拌過夜，反應結束後，乙酸乙酯萃取，飽和食鹽水洗，合併有機相，乾燥濃縮後得到粗品，再經製備HPLC純化得到2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)- N,2-二甲基丙醯胺Cpd-54(100 mg)，收率49%。 DMF (200 mg, 0.36 mmol) was added to 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid Cpd-54b ( 200 mg, 0.36 mmol), methylamine hydrochloride (33 mg, 0.4 mmol), 2-(7-azabenzotriazol-1-yl) -N , N , N ', N' -tetramethyluronium hexafluorophosphate (140 mg, 0.36 mmol), and triethylamine (0.1 mL, 0.72 mmol). mL), the reaction solution was stirred at room temperature overnight. After the reaction was completed, it was extracted with ethyl acetate and washed with saturated brine. The organic phases were combined, dried and concentrated to obtain a crude product, which was then purified by preparative HPLC to obtain 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl) -N ,2-dimethylpropionamide Cpd-54 (100 mg), with a yield of 49%.

MS m/z(ESI)：571.2(M+1)。MS m/z(ESI):571.2(M+1).

HPLC：98.87%(214 nm), 98.76%(254 nm)。HPLC: 98.87%(214 nm), 98.76%(254 nm).

¹H NMR(400 MHz, DMSO)δ 8.69(s, 1H), 8.60(d, J= 2.2 Hz, 1H), 8.14–8.02(m, 1H), 7.87(s, 1H), 7.76(d, J= 7.3 Hz, 1H), 7.47(t, J= 7.7 Hz, 1H), 7.41(d, J= 7.4 Hz, 2H), 6.88(s, 1H), 5.50(s, 2H), 2.54(d, J= 4.4 Hz, 3H), 2.10(s, 3H), 2.05(d, J= 6.7 Hz, 3H), 1.48(s, 6H). ¹ H NMR (400 MHz, DMSO) δ 8.69(s, 1H), 8.60(d, J = 2.2 Hz, 1H), 8.14–8.02(m, 1H), 7.87(s, 1H), 7.76(d, J = 7.3 Hz, 1H), 7.47(t, J = 7.7 Hz, 1H), 7.41(d, J = 7.4 Hz, 2H), 6.88(s, 1H), 5.50(s, 2H), 2.54(d, J = 4.4 Hz, 3H), 2.10(s, 3H), 2.05(d, J = 6.7 Hz, 3H), 1.48(s, 6H).

化合物Cpd-54經掌性拆分得到Cpd-54A(29.06 mg, t _R= 2.074 min)以及Cpd-54B(26.03 mg, t _R= 3.600 min)。 Compound Cpd-54 was subjected to chiral separation to obtain Cpd-54A (29.06 mg, t _R = 2.074 min) and Cpd-54B (26.03 mg, t _R = 3.600 min).

Cpd-54ACpd-54A

MS m/z(ESI)：571.1(M+1)。MS m/z(ESI):571.1(M+1).

HPLC：100%(214 nm), 99.56%(254 nm)。HPLC: 100% (214 nm), 99.56% (254 nm).

¹H NMR(400 MHz, DMSO)δ 8.68(s, 1H), 8.59(d, J= 2.3 Hz, 1H), 8.08(td, J= 9.9, 2.3 Hz, 1H), 7.86(s, 1H), 7.76(d, J= 7.4 Hz, 1H), 7.46(t, J= 7.7 Hz, 1H), 7.41(d, J= 8.1 Hz, 2H), 6.87(s, 1H), 5.50(s, 2H), 2.53(d, J= 4.5 Hz, 3H), 2.10(s, 3H), 2.04(s, 3H), 1.47(s, 6H). ¹ H NMR (400 MHz, DMSO) δ 8.68(s, 1H), 8.59(d, J = 2.3 Hz, 1H), 8.08(td, J = 9.9, 2.3 Hz, 1H), 7.86(s, 1H), 7.76(d, J = 7.4 Hz, 1H), 7.46(t, J = 7.7 Hz, 1H), 7.41(d, J = 8.1 Hz, 2H), 6.87(s, 1H), 5.50(s, 2H), 2.53(d, J = 4.5 Hz, 3H), 2.10(s, 3H), 2.04(s, 3H), 1.47(s, 6H).

Cpd-54BCpd-54B

MS m/z(ESI)：571.1(M+1)。MS m/z(ESI):571.1(M+1).

HPLC：99.38%(214 nm), 98.78%(254 nm)。HPLC: 99.38%(214 nm), 98.78%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.70(s, 1H), 8.61(d, J= 2.3 Hz, 1H), 8.10(td, J= 9.9, 2.3 Hz, 1H), 7.88(s, 1H), 7.77(d, J= 7.5 Hz, 1H), 7.48(t, J= 7.7 Hz, 1H), 7.43(d, J= 8.0 Hz, 2H), 6.89(s, 1H), 5.52(s, 2H), 2.55(d, J= 4.5 Hz, 3H), 2.12(s, 3H), 2.06(s, 3H), 1.49(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.70(s, 1H), 8.61(d, J = 2.3 Hz, 1H), 8.10(td, J = 9.9, 2.3 Hz, 1H), 7.88(s, 1H), 7.77(d, J = 7.5 Hz, 1H), 7.48(t, J = 7.7 Hz, 1H), 7.43(d, J = 8.0 Hz, 2H), 6.89(s, 1H), 5.52(s, 2H), 2.55(d, J = 4.5 Hz, 3H), 2.12(s, 3H), 2.06(s, 3H), 1.49(s, 6H).

實施例8Embodiment 8

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-1-(5-(6-(2-羥基丙烷-2-基)吡啶-2-基)-2-甲基苯基)-6-甲基吡啶-2(1H)-酮 3-Chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-1-(5-(6-(2-hydroxypropane-2-yl)pyridin-2-yl)-2-methylphenyl)-6-methylpyridin-2(1H)-one

第一步first step

稱取化合物Cpd-45f(500mg，1.1mmol)、聯硼酸頻那醇酯(558.5mg，2.2mmol)、醋酸鉀(382.2mg，3.3mmol)和催化量的Pd(dppf)Cl ₂置於100ml的茄型瓶中，加入二氧六環(50ml)，加熱至100度，反應4小時後，冷卻至室溫，過濾，濃縮，水洗，乙酸乙酯萃取，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到產品Cpd-55a(496.5mg)，收率90%。 Compound Cpd-45f (500 mg, 1.1 mmol), pinacol diborate (558.5 mg, 2.2 mmol), potassium acetate (382.2 mg, 3.3 mmol) and a catalytic amount of Pd(dppf) _Cl2 were weighed and placed in a 100 ml eggplant-shaped bottle, and dioxane (50 ml) was added. The mixture was heated to 100 degrees. After reacting for 4 hours, it was cooled to room temperature, filtered, concentrated, washed with water, extracted with ethyl acetate, dried and concentrated, and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain the product Cpd-55a (496.5 mg) with a yield of 90%.

MS m/z(ESI)：503.17(M+1)。MS m/z(ESI):503.17(M+1).

第二步Step 2

稱取化合物Cpd-55a(300mg，0.6mmol)和化合物Cpd-55b(141.7mg，0.66mmol)、四三苯基膦鈀(65 mg, 0.056 mmol)以及碳酸鈉(119 mg, 1.12 mmol)中加入四氫呋喃/水=2/1(6 mL)，反應液在90攝氏度下攪拌1小時，反應結束後，乙酸乙酯萃取，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到粗產品，再經製備HPLC純化(FA，流動相：ACN：H ₂O(0.1% FA)=49%：51%)得到3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-1-(5-(6-(2-羥基丙烷-2-基)吡啶-2-基)-2-甲基苯基)-6-甲基吡啶-2(1H)-酮 Cpd-55(169mg)，收率55%。 Compound Cpd-55a (300 mg, 0.6 mmol) and compound Cpd-55b (141.7 mg, 0.66 mmol), tetrakistriphenylphosphine palladium (65 mg, 0.056 mmol) and sodium carbonate (119 mg, 1.12 mmol) were weighed and tetrahydrofuran/water = 2/1 (6 mL). The reaction solution was stirred at 90 degrees Celsius for 1 hour. After the reaction was completed, it was extracted with ethyl acetate, dried and concentrated, and then separated and purified by silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain a crude product, which was then purified by preparative HPLC (FA, mobile phase: ACN: H ₂ O (0.1% FA) = 49%: 51%) to give 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-1-(5-(6-(2-hydroxypropane-2-yl)pyridin-2-yl)-2-methylphenyl)-6-methylpyridin-2(1H)-one Cpd-55 (169 mg) with a yield of 55%.

MS m/z(ESI)：512.17(M+1)。MS m/z(ESI):512.17(M+1).

¹H NMR(400 MHz, CDCl3)δ 8.41(d, J = 2.4 Hz, 1H), 8.00 – 7.90(m, 2H), 7.81(s, 1H), 7.69(d, J = 7.2 Hz, 1H), 7.50(d, J = 8.0 Hz, 1H), 7.41(d, J = 7.0 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J = 1.6 Hz, 2H), 2.15(s, 3H), 2.08(s, 3H), 1.65(d, J = 8.0 Hz, 6H). ¹ H NMR (400 MHz, CDCl3) δ 8.41(d, J = 2.4 Hz, 1H), 8.00 – 7.90(m, 2H), 7.81(s, 1H), 7.69(d, J = 7.2 Hz, 1H), 7.50(d, J = 8.0 Hz, 1H), 7.41(d, J = 7.0 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J = 1.6 Hz, 2H), 2.15(s, 3H), 2.08(s, 3H), 1.65(d, J = 8.0 Hz, 6H).

化合物Cpd-55經掌性拆分得到Cpd-55A(78 mg, t _R= 1.023 min)和Cpd-55B(68 mg, t _R= 1.398 min)。 Compound Cpd-55 was separated by chiral separation to obtain Cpd-55A (78 mg, t _R = 1.023 min) and Cpd-55B (68 mg, t _R = 1.398 min).

Cpd-55ACpd-55A

MS m/z(ESI)：512.2(M+1)。MS m/z(ESI):512.2(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.41(d, J= 2.4 Hz, 1H), 7.96(d, J= 8.0 Hz, 2H), 7.81(s, 1H), 7.70(d, J= 7.8 Hz, 1H), 7.51(d, J= 8.0 Hz, 1H), 7.43(d, J= 7.8 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J= 1.8Hz, 2H), 2.15(s, 3H), 2.09(s, 3H), 1.66(d, J= 8.0 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.41(d, J = 2.4 Hz, 1H), 7.96(d, J = 8.0 Hz, 2H), 7.81(s, 1H), 7.70(d, J = 7.8 Hz, 1H), 7.51(d, J = 8.0 Hz, 1H), 7.43(d, J = 7.8 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J = 1.8Hz, 2H), 2.15(s, 3H), 2.09(s, 3H), 1.66(d, J = 8.0 Hz, 6H).

Cpd-55BCpd-55B

MS m/z(ESI)：512.2(M+1)。MS m/z(ESI):512.2(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.41(d, J= 2.4 Hz, 1H), 7.98(dd, J= 8.0, 1.6 Hz, 1H), 7.88(t, J= 7.8 Hz, 1H), 7.80(d, J= 1.6 Hz, 1H), 7.67(d, J= 7.8Hz, 1H), 7.49(d, J= 8.0 Hz, 1H), 7.38(d, J= 7.8 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J= 1.8 Hz, 2H), 2.14(s, 3H), 2.05(s, 3H), 1.63(d, J= 8.4 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.41(d, J = 2.4 Hz, 1H), 7.98(dd, J = 8.0, 1.6 Hz, 1H), 7.88(t, J = 7.8 Hz, 1H), 7.80(d, J = 1.6 Hz, 1H), 7.67(d, J = 7.8Hz, 1H), 7.49(d, J = 8.0 Hz, 1H), 7.38(d, J = 7.8 Hz, 1H), 7.34 – 7.29(m, 1H), 6.36(s, 1H), 5.41(d, J = 1.8 Hz, 2H), 2.14(s, 3H), 2.05(s, 3H), 1.63(d, J = 8.4 Hz, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALCEL OD-3，4.6mm*150mm, 3μm；流動相：CO ₂/MeOH(0.1%DEA)=70/30；柱溫：37度。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation column model: Daicel CHIRALCEL OD-3, 4.6mm*150mm, 3μm; Mobile phase: CO ₂ /MeOH (0.1%DEA) = 70/30; Column temperature: 37 degrees.

實施例9Embodiment 9

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基- 2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙醯胺 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropionamide

第一步first step

向2',3-二氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53b(1 g, 2.32 mmol)、2-甲基-2-(3-(4,4,5,5-四甲基-1,3,2-二氧雜硼雜環戊烷-2-基)苯基)丙酸乙酯(884 mg, 2.78 mmol)、四三苯基膦鈀(268 mg, 0.232 mmol)以及碳酸鉀(640 mg, 4.64 mmol)中加入四氫呋喃/水=2/1(15 mL)，反應液在90攝氏度下攪拌1小時，反應結束後，乙酸乙酯萃取，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=1/2)分離純化得到2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸乙酯Cpd-37a(1.2 g)，收率88%。 To 2',3-dichloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-53b (1 g, 2.32 mmol), ethyl 2-methyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate (884 mg, 2.78 mmol), palladium tetrakistriphenylphosphine (268 mg, 0.232 mmol) and potassium carbonate (640 mg, 4.64 mmol) was added tetrahydrofuran/water = 2/1 (15 The reaction solution was stirred at 90 °C for 1 h. After the reaction was completed, the product was extracted with ethyl acetate, dried, concentrated, and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/2) to obtain 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid ethyl ester Cpd-37a (1.2 g) in a yield of 88%.

MS m/z(ESI)：586.2(M+1)。MS m/z(ESI):586.2(M+1).

第二步Step 2

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸的製備 Preparation of 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoic acid

向2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙酸乙酯Cpd-37a(600 mg, 1.02 mmol)以及氫氧化鋰(245 mg, 10.2 mmol)中加入異丙醇：四氫呋喃：水=1:1:1(30 mL)，反應液在50攝氏度下攪拌過夜，反應結束後，旋乾，加DMF，抽濾，濾液粗品Cpd-37b直接用於下一步。 Isopropanol: tetrahydrofuran: water = 1:1:1 (30 mL) was added to ethyl 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanoate Cpd-37a (600 mg, 1.02 mmol) and lithium hydroxide (245 mg, 10.2 mmol). The reaction solution was stirred at 50 °C overnight. After the reaction was completed, the solution was dried by spin drying, DMF was added, and the solution was filtered. The crude filtrate Cpd-37b was directly used in the next step.

MS m/z(ESI)：558.1(M+1)。MS m/z(ESI):558.1(M+1).

第三步third step

2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基- 2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙醯胺的製備 Preparation of 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropionamide

向上一步的粗品Cpd-37b的DMF溶液中加入氯化銨(54 mg, 1.02 mmol)、HATU(388 mg, 1.02 mmol)以及三乙胺(206 mg, 2.04 mmol)，反應液在室溫下攪拌1小時，反應結束後，直接經製備得到2-(3-(3-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基- 2-側氧基-2 H-[1,4'-聯吡啶]-2'-基)苯基)-2-甲基丙醯胺Cpd-37(122 mg)，收率22%。 To the DMF solution of the crude product Cpd-37b in the previous step were added ammonium chloride (54 mg, 1.02 mmol), HATU (388 mg, 1.02 mmol) and triethylamine (206 mg, 2.04 mmol), and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, 2-(3-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2-oxo- 2H- [1,4'-bipyridyl]-2'-yl)phenyl)-2-methylpropanamide Cpd-37 (122 mg) was directly prepared with a yield of 22%.

MS m/z(ESI)：557.1(M+1)。MS m/z(ESI):557.1(M+1).

HPLC：99.00%(214 nm), 99.22%(254 nm)。HPLC: 99.00%(214 nm), 99.22%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.55(s, 1H), 8.40(d, J= 2.1 Hz, 1H), 7.99(s, 1H), 7.82(d, J= 6.2 Hz, 1H), 7.68 – 7.63(m, 1H), 7.58 – 7.51(m, 1H), 7.35 – 7.29(m, 1H), 6.45(s, 1H), 5.42(d, J= 1.4 Hz, 2H), 5.33(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.63(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.55(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 7.99(s, 1H), 7.82(d, J = 6.2 Hz, 1H), 7.68 – 7.63(m, 1H), 7.58 – 7.51(m, 1H), 7.35 – 7.29(m, 1H), 6.45(s, 1H), 5.42(d, J = 1.4 Hz, 2H), 5.33(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.63(s, 6H).

化合物Cpd-37經掌性拆分得到Cpd-37A(t _R= 3.398 min)(47.45 mg)以及Cpd-37B(t _R= 4.164 min)(45.4 mg)。 Compound Cpd-37 was subjected to chiral separation to obtain Cpd-37A (t _R = 3.398 min) (47.45 mg) and Cpd-37B (t _R = 4.164 min) (45.4 mg).

Cpd-37BCpd-37B

MS m/z(ESI)：557.1(M+1)。MS m/z(ESI):557.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.57(s, 1H), 8.40(d, J= 2.3 Hz, 1H), 8.01(s, 1H), 7.87 – 7.80(m, 1H), 7.48(d, J= 6.3 Hz, 2H), 7.36 – 7.29(m, 1H), 6.44(s, 1H), 5.42(d, J= 1.6 Hz, 2H), 5.27(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.64(d, J= 2.6 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.57(s, 1H), 8.40(d, J = 2.3 Hz, 1H), 8.01(s, 1H), 7.87 – 7.80(m, 1H), 7.48(d, J = 6.3 Hz, 2H), 7.36 – 7.29(m, 1H), 6.44(s, 1H), 5.42(d, J = 1.6 Hz, 2H), 5.27(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.64(d, J = 2.6 Hz, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：REGIS(S,S)-Whelk O1, 250mm*30mm I.D., 10μm；流動相：CO ₂/MeOH[0.2%NH ₃(7M Solution in MeOH)]=70/30；柱溫：37度。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: REGIS (S, S)-Whelk O1, 250mm*30mm ID, 10μm; Mobile phase: CO ₂ /MeOH [0.2% NH ₃ (7M Solution in MeOH)] = 70/30; Column temperature: 37 degrees.

實施例10Embodiment 10

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(2-(2-羥基丙-2-基)嘧啶-4-基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮 3-Chloro-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-1-(5-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-2-methylphenyl)-6-methylpyridin-2(1 H )-one

第一步first step

1-(5-溴-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-bromo-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one

將1-(5-溴-2-甲基苯基)-4-羥基-6-甲基吡啶-2(1 H)-酮 Cpd-45d(1 g，3 mmol)、2-(氯甲基- D ₂)-3,5-二氟吡啶 Cpd-40i(0.59 g，3 mol)溶於N,N-二甲基甲醯胺(20 mL)，隨後加入18-冠醚-6(0.09 g，0.3 mol)以及碳酸鉀(0.94 g，6 mol)，反應液在60攝氏度下攪拌1小時，反應結束後，加入水淬滅，用乙酸乙酯萃取(3 × 10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=9/1)純化後得到1-(5-溴-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮 Cpd-56a(1.3 g)，產率：82.35%。 1-(5-Bromo-2-methylphenyl)-4 _- hydroxy-6-methylpyridin-2( 1H )-one Cpd-45d (1 g, 3 mmol) and 2-(chloromethyl- D2 )-3,5-difluoropyridine Cpd-40i (0.59 g, 3 mol) were dissolved in N,N-dimethylformamide (20 mL), followed by the addition of 18-crown-6 (0.09 g, 0.3 mol) and potassium carbonate (0.94 g, 6 mol). The reaction solution was stirred at 60 °C for 1 hour. After the reaction was completed, water was added to quench the reaction and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 9/1) to obtain 1-(5-bromo-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56a (1.3 g) with a yield of 82.35%.

MS m/z(ESI)：423.0(M+1)。MS m/z (ESI): 423.0 (M+1).

第二步Step 2

1-(5-乙醯基-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-acetyl-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one

將1-(5-溴-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮 Cpd-56a(1.3 g，3 mmol)、三丁基(1-乙氧基乙烯基)錫烷(1.68 g，4 mmol)和雙三苯基磷二氯化鈀(0.22 g，0.3 mmol)溶解在1,4-二氧六環(20 mL)中，在130度攪拌16小時。反應結束後所得深色溶液通過矽藻土過濾，用乙酸乙酯漂洗。濃縮濾液，將殘餘物溶於四氫呋喃(20 mL)和濃鹽酸(1 mL)中。將反應混合物在25度下攪拌1小時。反應結束後，用碳酸氫鈉淬滅反應，用水(20 mL)洗滌，用乙酸乙酯(3×10 mL)萃取。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)純化後得到1-(5-乙醯基-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮 Cpd-56b(1.5 g)，產率：93.55%。 1-(5-Bromo-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56a (1.3 g, 3 mmol), tributyl(1-ethoxyvinyl)tinane (1.68 g, 4 mmol) and bistriphenylphosphine dichloride (0.22 g, 0.3 mmol) were dissolved in 1,4-dioxane (20 mL) and stirred at 130 degrees for 16 hours. After the reaction was completed, the dark solution was filtered through diatomaceous earth and rinsed with ethyl acetate. The filtrate was concentrated and the residue was dissolved in tetrahydrofuran (20 mL) and concentrated hydrochloric acid (1 mL). The reaction mixture was stirred at 25 degrees for 1 hour. After the reaction was completed, the reaction was quenched with sodium bicarbonate, washed with water (20 mL), and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 1-(5-acetyl-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56b (1.5 g) with a yield of 93.55%.

MS m/z(ESI)：387.1(M+1)。MS m/z(ESI):387.1(M+1).

第三步third step

1-(5-乙醯基-2-甲基苯基)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 1-(5-acetyl-2-methylphenyl)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one

向 1-(5-乙醯基-2-甲基苯基)-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮Cpd-56b(1.5 g，3 mmol)、N-溴代丁二醯亞胺(0.57 g， 4 mmol)、二氯乙酸(0.05 g，0.3 mmol)加入異丙醇(20 mL)，反應在60攝氏度下反應3小時，反應結束後，加入水淬滅，用乙酸乙酯萃取(3 × 10 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)，純化後得到1-(5-乙醯基-2-甲基苯基)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮 Cpd-56c(680 mg)，產率：38.46%。 Isopropanol (20 mL) was added to 1-(5-acetyl-2-methylphenyl)-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56b (1.5 g, 3 mmol), N-bromosuccinimide (0.57 g, 4 mmol) and dichloroacetic acid (0.05 g, 0.3 mmol). The reaction was carried out at 60 °C for 3 h. After the reaction was completed, water was added to quench the reaction and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 1-(5-acetyl-2-methylphenyl)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56c (680 mg) in a yield of 38.46%.

MS m/z(ESI)：421.0(M+1)。MS m/z(ESI):421.0(M+1).

第四步the fourth step

( E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(3-(二甲胺基)丙烯醯基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of ( E )-3-chloro-4-((3,5-difluoropyridin- ₂ -yl)methoxy-( D2 )-1-(5-(3-(dimethylamino)acryloyl)-2-methylphenyl)-6-methylpyridin-2( 1H )-one

將1-(5-乙醯基-2-甲基苯基)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-6-甲基吡啶-2(1 H)-酮Cpd-56c(1.24 g，2 mmol)溶解在N,N-二甲基甲醯胺(15 mL)中，加入N,N-二甲基甲醯胺二甲基縮醛(1.38 g，11 mmol)。將反應混合物在55度下攪拌16小時。反應結束後，用水淬滅反應，用乙酸乙酯(3×10 mL)萃取。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=5/1)，得到( E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(3-(二甲胺基)丙烯醯基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮 Cpd-56d(1 g)，產率：65.52%。 1-(5-Acetyl-2-methylphenyl)-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-6-methylpyridin-2(1 H )-one Cpd-56c (1.24 g, 2 mmol) was dissolved in N,N-dimethylformamide (15 mL), and N,N-dimethylformamide dimethyl acetal (1.38 g, 11 mmol) was added. The reaction mixture was stirred at 55 degrees for 16 hours. After the reaction was completed, the reaction was quenched with water and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain ( E )-3-chloro-4-((3,5-difluoropyridin- ₂ -yl)methoxy- D2 )-1-(5-(3-(dimethylamino)acryloyl)-2-methylphenyl)-6-methylpyridin-2( 1H )-one Cpd-56d (1 g) in a yield of 65.52%.

MS m/z(ESI)：476.1(M+1)。MS m/z(ESI):476.1(M+1).

第五步the fifth step

3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(2-(2-羥基丙-2-基)嘧啶-4-基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮的製備 Preparation of 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy- D ₂ )-1-(5-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-2-methylphenyl)-6-methylpyridin-2(1 H )-one

將( E)-3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(3-(二甲胺基)丙烯醯基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮 Cpd-56d(130 mg，0.27 mmol)溶解在N,N-二甲基甲醯胺(5 mL)中，加入2-羥基-2-甲基丙脒Cpd-56e(83 mg，0.81 mmol)、碳酸鉀(151 mg，1.09 mmol)。將反應混合物在60度下攪拌16小時。反應結束後，用水淬滅反應，用乙酸乙酯(3×10 mL)萃取。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮。濃縮液通過製備純化並凍乾得到3-氯-4-((3,5-二氟吡啶-2-基)甲氧基- D ₂)-1-(5-(2-(2-羥基丙-2-基)嘧啶-4-基)-2-甲基苯基)-6-甲基吡啶-2(1 H)-酮Cpd-56(2.35 mg)，產率：1.50%。 ( E )-3-Chloro-4-((3,5-difluoropyridin- ₂ -yl)methoxy- D2 )-1-(5-(3-(dimethylamino)acryloyl)-2-methylphenyl)-6-methylpyridin-2( 1H )-one Cpd-56d (130 mg, 0.27 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 2-hydroxy-2-methylpropionamidine Cpd-56e (83 mg, 0.81 mmol) and potassium carbonate (151 mg, 1.09 mmol) were added. The reaction mixture was stirred at 60 degrees for 16 hours. After the reaction was completed, the reaction was quenched with water and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was purified by preparative purification and lyophilized to give 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy _- D2 )-1-(5-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-2-methylphenyl)-6-methylpyridin-2( 1H )-one Cpd-56 (2.35 mg), yield: 1.50%.

MS m/z(ESI)：515.1(M+1)。MS m/z(ESI):515.1(M+1).

HPLC：90.05%(214 nm), 90.05%(254 nm).HPLC: 90.05%(214 nm), 90.05%(254 nm).

¹H NMR(400 MHz, CDCl ₃- d6)δ 8.76(d, J= 5.4 Hz, 1H), 8.41(d, J= 2.2 Hz, 1H), 8.10(d, J= 8.1 Hz, 1H), 7.88(s, 1H), 7.56(d, J= 5.4 Hz, 1H), 7.52(d, J= 8.1 Hz, 1H), 7.35-7.30(m, 1H), 6.38(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.64(d, J= 3.8 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ - d6 )δ 8.76(d, J = 5.4 Hz, 1H), 8.41(d, J = 2.2 Hz, 1H), 8.10(d, J = 8.1 Hz, 1H), 7.88(s, 1H), 7.56(d, J = 5.4 Hz, 1H), 7.52(d, J = 8.1 Hz, 1H), 7.35-7.30(m, 1H), 6.38(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.64(d, J = 3.8 Hz, 6H).

化合物Cpd-56經掌性拆分得到Cpd-56A(10.08 mg, t _R= 1.031 min)和Cpd-56B(4.51 mg, t _R= 2.208 min)。 Compound Cpd-56 was subjected to chiral separation to obtain Cpd-56A (10.08 mg, t _R = 1.031 min) and Cpd-56B (4.51 mg, t _R = 2.208 min).

Cpd-56ACpd-56A

MS m/z(ESI)：515.1(M+1)。MS m/z(ESI):515.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.76(d, J= 5.4 Hz, 1H), 8.41(d, J= 2.3 Hz, 1H), 8.11(d, J= 8.0 Hz, 1H), 7.89(s, 1H), 7.60(d, J= 5.3 Hz, 1H), 7.52(d, J= 8.1 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.65(d, J= 3.5 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.76(d, J = 5.4 Hz, 1H), 8.41(d, J = 2.3 Hz, 1H), 8.11(d, J = 8.0 Hz, 1H), 7.89(s, 1H), 7.60(d, J = 5.3 Hz, 1H), 7.52(d, J = 8.1 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.65(d, J = 3.5 Hz, 6H).

Cpd-56BCpd-56B

MS m/z(ESI)：515.2(M+1)。MS m/z (ESI): 515.2 (M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.74(d, J= 5.2 Hz, 1H), 8.41(d, J= 2.3 Hz, 1H), 8.11 – 8.08(m, 1H), 7.88(d, J= 1.3 Hz, 1H), 7.56(d, J= 5.2 Hz, 1H), 7.51(d, J= 8.0 Hz, 1H), 7.35 – 7.30(m, 1H), 6.38(s, 1H), 2.16(s, 3H), 1.98(s, 3H), 1.64(d, J= 4.0 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.74(d, J = 5.2 Hz, 1H), 8.41(d, J = 2.3 Hz, 1H), 8.11 – 8.08(m, 1H), 7.88(d, J = 1.3 Hz, 1H), 7.56(d, J = 5.2 Hz, 1H), 7.51(d, J = 8.0 Hz, 1H), 7.35 – 7.30(m, 1H), 6.38(s, 1H), 2.16(s, 3H), 1.98(s, 3H), 1.64(d, J = 4.0 Hz, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALCEL OD, 250mm * 30mm I.D.，10μm；流動相：CO ₂/MeOH[0.2%NH ₃(7M Solution in MeOH)]= 50/50。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation column model: Daicel CHIRALCEL OD, 250mm * 30mm ID, 10μm; Mobile phase: CO ₂ /MeOH [0.2% NH ₃ (7M Solution in MeOH)] = 50/50.

實施例11Embodiment 11

3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-6''-(2-羥基丙-2-基)-5',6-二甲基-2H -[1,4':2',2''-三聯吡啶]-2-酮 3-Bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-6''-(2-hydroxypropan-2-yl)-5',6-dimethyl-2H-[1,4':2',2''-terpyridine]-2-one

第一步first step

1-(3,5-二氟吡啶-2-基)乙-1-醇的製備Preparation of 1-(3,5-difluoropyridin-2-yl)ethan-1-ol

將1-(3,5-二氟吡啶-2-基)乙-1-酮 Cpd-67d(1.0 g, 6.4mmol)溶於甲醇中(10 mL)，隨後緩慢加入硼氫化鈉(0.06 g, 1.6mmol)，反應混合物在零下10攝氏度下攪拌0.5小時，反應結束後，用飽和氯化銨對反應液進行洗滌，並用乙酸乙酯萃取，隨後對有機相進行乾燥、過濾、濃縮後經矽膠管柱層析純化(流動相：乙酸乙酯/二氯甲烷 =20%-50%)，得到1-(3,5-二氟吡啶-2-基)乙-1-醇Cpd-67e(0.7 g)，產率：65%。1-(3,5-difluoropyridin-2-yl)ethan-1-one Cpd-67d (1.0 g, 6.4 mmol) was dissolved in methanol (10 mL), and then sodium borohydride (0.06 g, 1.6 mmol) was slowly added. The reaction mixture was stirred at -10 °C for 0.5 h. After the reaction was completed, the reaction solution was washed with saturated ammonium chloride and extracted with ethyl acetate. The organic phase was then dried, filtered, concentrated, and purified by silica gel column chromatography (mobile phase: ethyl acetate/dichloromethane = 20%-50%) to obtain 1-(3,5-difluoropyridin-2-yl)ethan-1-ol Cpd-67e (0.7 g) with a yield of 65%.

MS m/z(ESI)：160.1(M+1)。MS m/z (ESI): 160.1 (M+1).

第二步Step 2

2-(1-氯乙基)-3,5-二氟吡啶的製備Preparation of 2-(1-chloroethyl)-3,5-difluoropyridine

將1-(3,5-二氟吡啶-2-基)乙-1-醇Cpd-67e(500 mg, 3.14 mmol)溶於二氯甲烷(20 mL)，在0度下滴加二氯亞碸(1121 mg, 9.43mmol)，反應在50度下攪拌2小時。反應結束後，反應液濃縮得到粗品2-(1-氯乙基)-3,5-二氟吡啶Cpd-67f(500 mg)。產率：71.69%。1-(3,5-difluoropyridin-2-yl)ethan-1-ol Cpd-67e (500 mg, 3.14 mmol) was dissolved in dichloromethane (20 mL), and dichlorosulfonyl chloride (1121 mg, 9.43 mmol) was added dropwise at 0 degrees, and the reaction was stirred at 50 degrees for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain the crude product 2-(1-chloroethyl)-3,5-difluoropyridine Cpd-67f (500 mg). Yield: 71.69%.

MS m/z(ESI)：178.0(M+1)。MS m/z(ESI):178.0(M+1).

第三步third step

3-溴-2'-氯-3'-氟-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 3-bromo-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

將2'-氯-3'-氟-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-40e(500 mg, 1.861 mmol)溶於乙腈(10 mL)中，加入N-溴代丁二醯亞胺(331.22 mg, 1.861 mmol)、二氯乙酸(24 mg, 0.1861 mmol)。於60度反應2小時，反應結束後，加水和乙酸乙酯萃取，收集有機相，乾燥過濾富集，殘液經矽膠管柱(石油醚/乙酸乙酯=2/1)得到3-溴-2'-氯-3'-氟-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-67a(400 mg)。產率：58.75%。2'-Chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-40e (500 mg, 1.861 mmol) was dissolved in acetonitrile (10 mL), and N-bromosuccinimide (331.22 mg, 1.861 mmol) and dichloroacetic acid (24 mg, 0.1861 mmol) were added. The reaction was carried out at 60 degrees for 2 hours. After the reaction was completed, water and ethyl acetate were added for extraction. The organic phase was collected, dried, filtered and enriched. The residue was passed through a silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 3-bromo-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-67a (400 mg). Yield: 58.75%.

MS m/z(ESI)：347.0(M+1)。MS m/z (ESI): 347.0 (M+1).

第四步the fourth step

3-溴-2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2H-[1,4'-聯吡啶] -2-酮的製備Preparation of 3-bromo-2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

將3-溴-2'-氯-3'-氟-4-羥基-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-67a(400 mg, 1.1509 mmol)溶於N,N-二甲基甲醯胺(5 mL)中，加入2-(1-氯乙基)-3,5-二氟吡啶Cpd-67f(224.81 mg, 1.2660 mmol)、碳酸鉀(318.13 mg, 2.3018mmol)、18-冠醚-6(152.1 mg, 0.5755 mmol)。於60度反應16小時。反應結束後，加入水和乙酸乙酯萃取，收集有機相，乾燥過濾富集，殘液經矽膠管柱(石油醚/乙酸乙酯=1/1)純化得到3-溴-2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2H-[1,4'-聯吡啶] -2-酮Cpd-67b(190 mg)。產率：30.40%。3-Bromo-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-67a (400 mg, 1.1509 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 2-(1-chloroethyl)-3,5-difluoropyridine Cpd-67f (224.81 mg, 1.2660 mmol), potassium carbonate (318.13 mg, 2.3018 mmol), and 18-crown-6 (152.1 mg, 0.5755 mmol) were added. The mixture was reacted at 60 degrees for 16 hours. After the reaction was completed, water and ethyl acetate were added for extraction, the organic phase was collected, dried and filtered, and the residue was purified by silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain 3-bromo-2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-67b (190 mg). Yield: 30.40%.

MS m/z(ESI)：488.1(M+1)。MS m/z(ESI):488.1(M+1).

第五步the fifth step

3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2'-(三甲基甲錫烷基)-2H-[1,4'-聯吡啶]-2-酮的製備Preparation of 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2'-(trimethylmethanyl)-2H-[1,4'-bipyridinyl]-2-one

將3-溴-2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2H-[1,4'-聯吡啶] -2-酮Cpd-67b(190 mg, 0.3888 mmol)溶於1,4-二氧六環(5 mL)中，加入六甲基二錫(636.91 mg, 1.944 mmol)、三苯基砷(47.62 mg, 0.1555 mmol)、雙三苯基膦二氯化鈀(109.16 mg, 0.1555 mmol)。於100度封管反應兩小時。反應結束後，加入飽和硫代硫酸鈉和乙酸乙酯萃取，收集有機相，乾燥過濾富集，得粗品(Cpd-67c)直接用於下一步。3-Bromo-2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-67b (190 mg, 0.3888 mmol) was dissolved in 1,4-dioxane (5 mL), and hexamethyldistin (636.91 mg, 1.944 mmol), triphenylarsine (47.62 mg, 0.1555 mmol), and bistriphenylphosphinepalladium dichloride (109.16 mg, 0.1555 mmol) were added. The mixture was sealed and reacted at 100 degrees for two hours. After the reaction was completed, saturated sodium thiosulfate and ethyl acetate were added for extraction, and the organic phase was collected, dried, filtered, and enriched to obtain a crude product (Cpd-67c) which was directly used in the next step.

MS m/z(ESI)：618.1(M+1)。MS m/z(ESI):618.1(M+1).

第六步Step 6

3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-6''-(2-羥基丙-2-基)-5',6-二甲基-2H-[1,4':2',2''-三聯吡啶]-2-酮的製備Preparation of 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-6''-(2-hydroxypropan-2-yl)-5',6-dimethyl-2H-[1,4':2',2''-terpyridine]-2-one

將3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2'-(三甲基甲錫烷基)-2H-[1,4'-聯吡啶]-2-酮Cpd-67c(100 mg, 0.1621 mmol)溶於1,4-二氧六環中(5 mL)。加入2-(6-溴吡啶-2-基)丙-2-醇 Cpd-55b(105.08 mg, 0.4863 mmol)、雙三苯基膦二氯化鈀(22.76 mg, 0.0324 mmol)。於80度封管反應2小時，反應結束後，過濾富集，殘液經製備級管柱(乙腈/水=65/35)純化得到3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-6''-(2-羥基丙-2-基)-5',6-二甲基-2H-[1,4':2',2''-三聯吡啶]-2-酮Cpd-67(3.76 mg)。產率：3.95%。3-Bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2'-(trimethylmethanyl)-2H-[1,4'-bipyridinyl]-2-one Cpd-67c (100 mg, 0.1621 mmol) was dissolved in 1,4-dioxane (5 mL). 2-(6-bromopyridin-2-yl)propan-2-ol Cpd-55b (105.08 mg, 0.4863 mmol) and bis(triphenylphosphine)palladium dichloride (22.76 mg, 0.0324 mmol) were added. The reaction was carried out at 80°C for 2 hours. After the reaction was completed, the residue was filtered and concentrated, and the residue was purified by a preparative column (acetonitrile/water = 65/35) to obtain 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-6''-(2-hydroxypropan-2-yl)-5',6-dimethyl-2H-[1,4':2',2''-terpyridine]-2-one Cpd-67 (3.76 mg). Yield: 3.95%.

MS m/z(ESI)：589.1(M+1)。MS m/z(ESI):589.1(M+1).

HPLC：98.89%(214 nm), 97.88%(254 nm)。HPLC: 98.89%(214 nm), 97.88%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.56(s, 1H), 8.39(d, J= 2.4 Hz, 1H), 8.05(d, J= 7.7 Hz, 1H), 7.91(d, J= 8.1 Hz, 1H), 7.43(d, J= 7.7 Hz, 1H), 7.29(dd, J=10.0, 2.1 Hz, 1H), 6.13(s, 1H), 5.77(q, J= 6.6 Hz, 1H), 2.21(s, 3H), 1.99(d, J= 5.0 Hz, 3H), 1.87(d, J= 6.6 Hz, 3H), 1.57(d, J= 2.8 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56(s, 1H), 8.39(d, J = 2.4 Hz, 1H), 8.05(d, J = 7.7 Hz, 1H), 7.91(d, J = 8.1 Hz, 1H), 7.43(d, J = 7.7 Hz, 1H), 7.29(dd, J =10.0, 2.1 Hz, 1H), 6.13(s, 1H), 5.77(q, J = 6.6 Hz, 1H), 2.21(s, 3H), 1.99(d, J = 5.0 Hz, 3H), 1.87(d, J = 6.6 Hz, 3H), 1.57(d, J = 2.8 Hz, 6H).

實施例12Embodiment 12

3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 3-Chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

第一步first step

2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2'-氯-3'-氟-4-羥基-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-40e(4 g, 14.9 mmol)、2-(1-氯乙基)-3,5-二氟吡啶 Cpd-67f(3.97 g, 22.3 mmol)、18-冠醚-6(390 mg, 1.4 mmol)和碳酸鉀(6.18 g, 44.7 mmol)溶於N,N-二甲基甲醯胺(15 mL)，反應在60度下攪拌12小時。反應結束後，用水(20 mL)洗，用乙酸乙酯(3 × 20 mL)萃取，合併有機相，用飽和食鹽水(20 mL)洗，有機相用無水硫酸鈉乾燥，過濾，濃縮。粗產品用矽膠管柱(乙酸乙酯/石油醚=75%)純化得到2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-81a(3.7 g)，產率：54%。 2'-Chloro-3'-fluoro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-40e (4 g, 14.9 mmol), 2-(1-chloroethyl)-3,5-difluoropyridine Cpd-67f (3.97 g, 22.3 mmol), 18-crown-6 (390 mg, 1.4 mmol) and potassium carbonate (6.18 g, 44.7 mmol) were dissolved in N,N-dimethylformamide (15 mL) and the reaction was stirred at 60 degrees for 12 hours. After the reaction was completed, the mixture was washed with water (20 mL), extracted with ethyl acetate (3 × 20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 75%) to obtain 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-81a (3.7 g), yield: 54%.

MS m/z(ESI)：410.1(M+1)。MS m/z(ESI):410.1(M+1).

第二步Step 2

2',3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2',3-dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-81a(3.6 g, 8.8 mmol)和N-氯代丁二醯亞胺(1.29 g, 9.6 mmol)溶於N,N-二甲基甲醯胺(15 mL)，在60度下反應5小時。反應結束後，用硫代硫酸鈉水溶液(20 mL)淬滅，用乙酸乙酯(3 × 20 mL)萃取，合併有機相，用飽和食鹽水(20 mL)洗，有機相用無水硫酸鈉乾燥，過濾，濃縮。粗產品用矽膠管柱(乙酸乙酯/石油醚=80%)純化得到2',3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 Cpd-81b(2.4 g)，產率：57%。 2'-Chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-81a (3.6 g, 8.8 mmol) and N-chlorobutanediimide (1.29 g, 9.6 mmol) were dissolved in N,N-dimethylformamide (15 mL) and reacted at 60 degrees for 5 hours. After the reaction was completed, it was quenched with sodium thiosulfate aqueous solution (20 mL), extracted with ethyl acetate (3 × 20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 80%) to give 2',3-dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-81b (2.4 g) with a yield of 57%.

MS m/z(ESI)：444.0(M+1)。MS m/z(ESI):444.0(M+1).

第三步third step

3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(3-(2-hydroxypropane-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2',3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-81b (2.4 g, 5.4 mmol)、(3-(2-羥基丙烷-2-基)苯基)硼酸Cpd-40m(1.17 g, 6.4 mmol)、1'-雙(二苯基膦基)二茂鐵氯化鈀(400 mg, 0.5 mmol)和碳酸鈉(1.49 g, 10.8 mmol)溶於1,4-二氧六環：水=5：1(24 mL)，在90度下攪拌2小時。反應結束後，過濾，濃縮。粗產品通過製備純化(FA，流動相：ACN：H ₂O(0.1% FA)=50%：50%)，得到3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(3-(2-羥基丙烷-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-81 (1.6 g)。產率：53.70%。 2',3-Dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-81b ( 2.4 g, 5.4 mmol), (3-(2-hydroxypropane-2-yl)phenyl)boronic acid Cpd-40m (1.17 g, 6.4 mmol), 1'-bis(diphenylphosphino)ferrocenepalladium chloride (400 mg, 0.5 mmol) and sodium carbonate (1.49 g, 10.8 mmol) were dissolved in 1,4-dioxane: water = 5:1 (24 mL) and stirred at 90 degrees for 2 hours. After the reaction was completed, the mixture was filtered and concentrated. The crude product was purified by preparative purification (FA, mobile phase: ACN: _H2O (0.1% FA) = 50%: 50%) to give 3-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(3-(2-hydroxypropane-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-81 ( 1.6 g). Yield: 53.70%.

化合物Cpd-81經第一次掌性拆分得到光學純的化合物Cpd-81A(97.30 mg, t _R= 1.532 min)和Cpd-81B(79.38 mg, t _R= 1.937 min)，以及異構物混合物Cpd-81M。 Compound Cpd-81 was subjected to the first chiral resolution to obtain optically pure compounds Cpd-81A (97.30 mg, t _R = 1.532 min) and Cpd-81B (79.38 mg, t _R = 1.937 min), as well as an isomer mixture Cpd-81M.

Cpd-81ACpd-81A

MS m/z(ESI)：544.1(M+1)。MS m/z(ESI):544.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

1H NMR(400 MHz, CDCl ₃) δ8.52(s, 1H), 8.37(d, J= 2.4 Hz, 1H), 8.05(d, J= 1.2 Hz, 1H), 7.77(dd, J= 7.6, 1.2 Hz, 1H), 7.57 – 7.54(m, 1H), 7.43(d, J= 7.8 Hz, 1H), 7.30 – 7.23(m, 1H), 6.15(s, 1H), 5.77(q, J= 6.6 Hz, 1H), 2.16(s, 3H), 1.93(s, 3H), 1.85(d, J= 6.4 Hz, 3H), 1.60(s, 6H). 1H NMR (400 MHz, CDCl ₃ ) δ 8.52(s, 1H), 8.37(d, J = 2.4 Hz, 1H), 8.05(d, J = 1.2 Hz, 1H), 7.77(dd, J = 7.6, 1.2 Hz, 1H), 7.57 – 7.54(m, 1H), 7.43(d, J = 7.8 Hz, 1H), 7.30 – 7.23(m, 1H), 6.15(s, 1H), 5.77(q, J = 6.6 Hz, 1H), 2.16(s, 3H), 1.93(s, 3H), 1.85(d, J = 6.4 Hz, 3H), 1.60(s, 6H).

Cpd-81BCpd-81B

MS m/z(ESI)：544.1(M+1)。MS m/z(ESI):544.1(M+1).

HPLC：97.79%(214 nm), 97.97%(254 nm)。HPLC: 97.79%(214 nm), 97.97%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.56(s, 1H), 8.43(d, J= 2.4 Hz, 1H), 8.10(s, 1H), 7.84(d, J= 7.6 Hz, 1H), 7.62(d, J= 7.6 Hz, 1H), 7.48(t, J= 7.8 Hz, 1H), 7.36 - 7.29(m, 1H), 6.19(s, 1H), 5.82(q, J= 6.6 Hz, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.89(d, J= 6.4 Hz, 3H), 1.66(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56(s, 1H), 8.43(d, J = 2.4 Hz, 1H), 8.10(s, 1H), 7.84(d, J = 7.6 Hz, 1H), 7.62(d, J = 7.6 Hz, 1H), 7.48(t, J = 7.8 Hz, 1H), 7.36 - 7.29(m, 1H), 6.19(s, 1H), 5.82(q, J = 6.6 Hz, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.89(d, J = 6.4 Hz, 3H), 1.66(s, 6H).

第一次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK IG_3, 3.0*150mm, 3μm；流動相：A/B：CO ₂/MeOH(0.1%DEA)=70/30；柱溫：37度。 Conditions for the first separation: Instrument brand: Waters Acquity UPCC; Preparation-grade column model: Daicel CHIRALPAK IG_3, 3.0*150mm, 3μm; Mobile phase: A/B: CO ₂ /MeOH(0.1%DEA)=70/30; Column temperature: 37 degrees.

Cpd-81M經第二次掌性拆分得到光學純的化合物Cpd-81C(38.78 mg, t _R= 1.926 min)和Cpd-81D(27.37 mg, t _R= 2.331 min)。 Cpd-81M was subjected to a second chiral resolution to afford optically pure compounds Cpd-81C (38.78 mg, t _R = 1.926 min) and Cpd-81D (27.37 mg, t _R = 2.331 min).

Cpd-81CCpd-81C

MS m/z(ESI)：544.2(M+1)。MS m/z(ESI):544.2(M+1).

HPLC：99.22%(214 nm), 98.70%(254 nm)。HPLC: 99.22%(214 nm), 98.70%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.50(s, 1H), 8.34(d, J = 2.4 Hz, 1H), 8.01(s, 1H), 7.74(d, J = 7.4 Hz, 1H), 7.52(d, J = 7.8 Hz, 1H), 7.39(t, J = 7.8 Hz, 1H), 7.27-7.22(m, 1H), 6.10(s, 1H), 5.73(q, J = 6.6 Hz, 1H), 2.13(s, 3H), 1.90(s, 3H), 1.82(d, J = 6.6 Hz, 3H), 1.57(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.50(s, 1H), 8.34(d, J = 2.4 Hz, 1H), 8.01(s, 1H), 7.74(d, J = 7.4 Hz, 1H), 7.52(d, J = 7.8 Hz, 1H), 7.39(t, J = 7.8 Hz, 1H), 7.27-7.22(m, 1H), 6.10(s, 1H), 5.73(q, J = 6.6 Hz, 1H), 2.13(s, 3H), 1.90(s, 3H), 1.82(d, J = 6.6 Hz, 3H), 1.57(s, 6H).

Cpd-81DCpd-81D

MS m/z(ESI)：544.1(M+1)。MS m/z(ESI):544.1(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.53(s, 1H), 8.39(d, J = 2.4 Hz, 1H), 8.07(s, 1H), 7.80(d, J = 7.6 Hz, 1H), 7.58(d, J = 7.8 Hz, 1H), 7.44(d, J = 7.8 Hz, 1H), 7.31-7.27(m, 1H), 6.15(s, 1H), 5.78(q, J = 6.6 Hz, 1H), 2.13(s, 3H), 1.95(s, 3H), 1.86(d, J = 6.6 Hz, 3H), 1.63(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.53(s, 1H), 8.39(d, J = 2.4 Hz, 1H), 8.07(s, 1H), 7.80(d, J = 7.6 Hz, 1H), 7.58(d, J = 7.8 Hz, 1H), 7.44(d, J = 7.8 Hz, 1H), 7.31-7.27(m, 1H), 6.15(s, 1H), 5.78(q, J = 6.6 Hz, 1H), 2.13(s, 3H), 1.95(s, 3H), 1.86(d, J = 6.6 Hz, 3H), 1.63(s, 6H).

第二次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK OJ_3, 3*150mm, 3μm；流動相：A/B：CO ₂/MeOH(0.1%DEA)=80/20；柱溫：37度。 Second splitting conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALPAK OJ_3, 3*150mm, 3μm; Mobile phase: A/B: CO ₂ /MeOH(0.1%DEA)=80/20; Column temperature: 37 degrees.

實施例13Embodiment 13

3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 3-Bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

第一步first step

2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-acetyl-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

氮氣保護下，向2'-氯-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-53a(2.0 g, 5 mmol)、三丁基(1-乙氧基乙烯基)錫烷(2.1 mL, 6 mmol)及雙四三苯基膦二氯化鈀(0.35 g, 0.5 mmol)中加入1,4-二氧六環(30 mL)，反應液在130攝氏度下攪拌12 h，待反應液冷卻至室溫後，加入氟化鉀溶液淬滅並攪拌30分鐘，然後過濾，濾液用乙酸乙酯萃取三次，濃縮後溶於四氫呋喃中(15 mL)，然後加入濃鹽酸(10 mL)，室溫下攪拌30分鐘，然後在冰浴下使用4 N氫氧化鈉中和反應液pH = 7，乙酸乙酯萃取三次，乾燥濃縮後經矽膠管柱(二氯甲烷/甲醇 = 99/1)分離純化得到2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65a(1.6 g)，收率79%。 Under nitrogen protection, 1,4-dioxane (30 mL) was added to 2'-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-53a (2.0 g, 5 mmol), tributyl(1-ethoxyvinyl)tinane (2.1 mL, 6 mmol) and bis(tetrakistriphenylphosphine)palladium dichloride (0.35 g, 0.5 mmol). The reaction solution was stirred at 130 °C for 12 h. After the reaction solution was cooled to room temperature, potassium fluoride solution was added to quench the reaction solution and stirred for 30 min. Then, the reaction solution was filtered and the filtrate was extracted three times with ethyl acetate. After concentration, it was dissolved in tetrahydrofuran (15 mL), then concentrated hydrochloric acid (10 mL) was added and stirred at room temperature for 30 min. Then, the reaction solution was neutralized to pH = 7 with 4 N sodium hydroxide in an ice bath, extracted three times with ethyl acetate, dried and concentrated, and then separated and purified by a silica gel column (dichloromethane/methanol = 99/1) to obtain 2'-acetyl-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-65a (1.6 g) in a yield of 79%.

MS m/z(ESI)：404.1(M+1)。MS m/z(ESI):404.1(M+1).

第二步Step 2

2'-乙醯基-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-acetyl-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2'-乙醯基-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65a(2.0 g, 5 mmol)以及NBS(0.89 g, 5 mmol)中加入 N, N-二甲基甲醯胺(30 mL)，反應液在60攝氏度下攪拌1小時，反應結束後，乾燥濃縮經矽膠管柱(石油醚/乙酸乙酯=3/7)分離純化得到2'-乙醯基-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65b(1.78 g)，收率74%。 N,N-dimethylformamide (30 mL) was added to 2'-acetyl-4-((3,5-difluoropyridin-2- yl )methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd -65a (2.0 g, 5 mmol) and NBS (0.89 g, 5 mmol). The reaction solution was stirred at 60 °C for 1 hour. After the reaction was completed, the product was dried, concentrated and purified by silica gel column (petroleum ether/ethyl acetate = 3/7) to obtain 2'-acetyl-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one. -[1,4'-Bipyridyl]-2-one Cpd-65b (1.78 g), yield 74%.

MS m/z(ESI)：482.0(M+1)。MS m/z(ESI):482.0(M+1).

第三步third step

( E)-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of ( E )-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向2'-乙醯基-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65b(1.0 g, 2.1 mmol)以及DMF-DMA(1.1 mL, 8.4 mmol)中加入 N, N-二甲基甲醯胺(20 mL)，反應液在55攝氏度下攪拌過夜，反應結束後，直接旋乾並經矽膠管柱(二氯甲烷/甲醇 = 95/5)分離純化得到( E)-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65c(1.07 g)，收率95%。 N,N-dimethylformamide ( 20 mL) was added to 2'-acetyl-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl] -2 - one Cpd-65b (1.0 g, 2.1 mmol) and DMF-DMA (1.1 mL, 8.4 mmol). The reaction solution was stirred at 55 °C overnight. After the reaction was completed, the product was directly spin-dried and purified by silica gel column (dichloromethane/methanol = 95/5) to obtain ( E )-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl- 2H-[1,4'-bipyridyl]-2-one. -[1,4'-Bipyridyl]-2-one Cpd-65c (1.07 g), yield 95%.

MS m/z(ESI)：537.0(M+1)。MS m/z(ESI):537.0(M+1).

第四步the fourth step

3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向( E)-3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-65c(500 mg, 0.93 mmol)、2-羥基-2-甲基丙脒(476 mg, 4.67 mmol)以及碳酸鉀(513 mg, 3.72 mmol)中加入 N, N-二甲基甲醯胺(5 mL)，反應液在55攝氏度下攪拌過夜，反應結束後，飽和食鹽水洗，乙酸乙酯萃取，有機相合併後乾燥濃縮並經矽膠管柱(二氯甲烷/甲醇 = 95/5)分離純化得到粗品，再經製備級管柱純化得到3-溴-4-((3,5-二氟吡啶-2-基)甲氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2H-[1,4'-聯吡啶]-2-酮Cpd-65(150 mg)，收率28%。 N,N-dimethylformamide (5 mL) was added to ( E )-3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5', 6 - dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-65c (500 mg, 0.93 mmol), 2-hydroxy-2-methylpropionamidine (476 mg, 4.67 mmol) and potassium carbonate (513 mg, 3.72 mmol). The reaction solution was stirred at 55 °C overnight. After the reaction was completed, the product was washed with saturated brine and extracted with ethyl acetate. The organic phases were combined, dried, concentrated and passed through a silica gel column (dichloromethane/methanol = 95/5) to obtain a crude product, which was then purified by a preparative column to obtain 3-bromo-4-((3,5-difluoropyridin-2-yl)methoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one Cpd-65 (150 mg) in a yield of 28%.

MS m/z(ESI)：576.0(M+1)。MS m/z (ESI): 576.0 (M+1).

HPLC：99.04%(214 nm), 98.55%(254 nm)。HPLC: 99.04%(214 nm), 98.55%(254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.02(d, J= 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J= 2.3 Hz, 1H), 8.09(td, J= 9.9, 2.3 Hz, 1H), 8.03(d, J= 5.2 Hz, 1H), 6.86(s,1H), 5.51(s, 2H), 5.07(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.02(d, J = 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J = 2.3 Hz, 1H), 8.09(td, J = 9.9, 2.3 Hz, 1H), 8.03(d, J = 5.2 Hz, 1H), 6.86(s,1H), 5.51(s, 2H), 5.07(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H).

化合物Cpd-65經掌性拆分得到Cpd-65A (t _R= 1.934 min, 40 mg)以及Cpd-65B (t _R= 3.070 min, 40 mg)。 Compound Cpd-65 was subjected to chiral separation to obtain Cpd-65A ( t _R = 1.934 min, 40 mg) and Cpd-65B ( t _R = 3.070 min, 40 mg).

Cpd-65ACpd-65A

MS m/z(ESI)：576.0(M+1)。MS m/z (ESI): 576.0 (M+1).

HPLC：98.24 %(214 nm), 100%(254 nm)。HPLC: 98.24% (214 nm), 100% (254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.02(d, J= 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J= 2.3 Hz, 1H), 8.10(td, J= 9.9, 2.3 Hz, 1H), 8.03(d, J= 5.2 Hz, 1H), 6.86(s,1H), 5.51(s, 2H), 5.08(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.02(d, J = 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J = 2.3 Hz, 1H), 8.10(td, J = 9.9, 2.3 Hz, 1H), 8.03(d, J = 5.2 Hz, 1H), 6.86(s,1H), 5.51(s, 2H), 5.08(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H).

Cpd-65BCpd-65B

MS m/z(ESI)：576.0(M+1)。MS m/z (ESI): 576.0 (M+1).

HPLC：99.57 %(214 nm), 99.47%(254 nm)。HPLC: 99.57%(214 nm), 99.47%(254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.01(d, J= 5.2 Hz, 1H), 8.76(s, 1H), 8.59(d, J= 2.2 Hz, 1H), 8.09(td, J= 10.0, 2.3 Hz, 1H), 8.02(d, J= 5.2 Hz, 1H), 6.85(s,1H), 5.50(s, 2H), 5.07(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.51(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.01(d, J = 5.2 Hz, 1H), 8.76(s, 1H), 8.59(d, J = 2.2 Hz, 1H), 8.09(td, J = 10.0, 2.3 Hz, 1H), 8.02(d, J = 5.2 Hz, 1H), 6.85(s,1H), 5.50(s, 2H), 5.07(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.51(s, 6H).

拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK AZ_3, 3*150mm, 3μm；流動相：CO ₂/MeOH(0.1% DEA)=60/40。 Separation conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALPAK AZ_3, 3*150mm, 3μm; Mobile phase: CO ₂ /MeOH (0.1% DEA) = 60/40.

實施例14Embodiment 14

3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮 3-Bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

第一步first step

2'-乙醯基-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

氮氣保護下，向2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-81a(2.8 g, 6.8 mmol)、三丁基(1-乙氧基乙烯基)錫烷(3.0 g, 8.2 mmol)及雙四三苯基膦二氯化鈀(0.48 g, 0.68 mmol)中加入1,4-二氧六環(30 mL)，反應液在130攝氏度下攪拌12 h，待反應液冷卻至室溫後，加入氟化鉀溶液淬滅並攪拌30分鐘，然後過濾，濾液用乙酸乙酯萃取三次，濃縮後溶於四氫呋喃中(20 mL)，然後加入濃鹽酸(10 mL)，室溫下攪拌30分鐘，然後在冰浴下使用4 N氫氧化鈉中和反應液pH = 7，乙酸乙酯萃取三次，乾燥濃縮後經矽膠管柱(石油醚/乙酸乙酯=35/65)分離純化得到2'-乙醯基-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68a(2.2 g)，收率77%。 Under nitrogen protection, 1,4-dioxane (30 mL) was added to 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-81a (2.8 g, 6.8 mmol), tributyl(1-ethoxyvinyl)tinane (3.0 g, 8.2 mmol) and bis(tetrakistriphenylphosphine)palladium dichloride (0.48 g, 0.68 mmol). The reaction solution was stirred at 130 °C for 12 h. After the reaction solution was cooled to room temperature, potassium fluoride solution was added to quench the reaction solution and stirred for 30 min. Then, the reaction solution was filtered and the filtrate was extracted three times with ethyl acetate. After concentration, it was dissolved in tetrahydrofuran (20 mL), then concentrated hydrochloric acid (10 mL) was added and stirred at room temperature for 30 min. Then, the reaction solution was neutralized to pH = 7 with 4 N sodium hydroxide in an ice bath, extracted three times with ethyl acetate, dried and concentrated, and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 35/65) to obtain 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-68a (2.2 g) with a yield of 77%.

MS m/z(ESI)：418.1(M+1)。MS m/z(ESI):418.1(M+1).

第二步Step 2

2'-乙醯基-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2'-乙醯基-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68a(1.5 g, 3.6 mmol)以及NBS(0.64 g, 3.6 mmol)中加入 N, N-二甲基甲醯胺(15 mL)，反應液在60攝氏度下攪拌1小時，反應結束後，乾燥濃縮經矽膠管柱(石油醚/乙酸乙酯=35/65)分離純化得到2'-乙醯基-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68b(1.35 g)，收率76%。 N,N-dimethylformamide (15 mL) was added to 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd -68a (1.5 g, 3.6 mmol) and NBS (0.64 g, 3.6 mmol). The reaction solution was stirred at 60 ° C for 1 hour. After the reaction was completed, the product was dried, concentrated and purified by silica gel column (petroleum ether/ethyl acetate = 35/65) to obtain 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one. -[1,4'-Bipyridyl]-2-one Cpd-68b (1.35 g), yield 76%.

MS m/z(ESI)：496.0(M+1)。MS m/z(ESI):496.0(M+1).

第三步third step

( E)-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of ( E )-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向2'-乙醯基-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68b(1.35 g, 2.7 mmol)以及DMF-DMA(1.4 mL, 10.9 mmol)中加入 N, N-二甲基甲醯胺(10 mL)，反應液在55攝氏度下攪拌過夜，反應結束後，直接旋乾並經矽膠管柱(二氯甲烷/甲醇 = 95/5)分離純化得到( E)-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68c(0.91 g)，收率67%。 N, N-dimethylformamide (10 mL) was added to 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl] -2 - one Cpd-68b (1.35 g, 2.7 mmol) and DMF-DMA (1.4 mL, 10.9 mmol). The reaction solution was stirred at 55 degrees Celsius overnight. After the reaction was completed, the product was directly spin-dried and purified by silica gel column (dichloromethane/methanol = 95/5) to obtain ( E )-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-68c (0.91 g), yield 67%.

MS m/z(ESI)：551.0(M+1)。MS m/z(ESI):551.0(M+1).

第四步the fourth step

3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5'-6-二甲基-2 H-[1,4'-聯吡啶]-2-酮的製備 Preparation of 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5'-6-dimethyl- 2H- [1,4'-bipyridyl]-2-one

向( E)-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2'-(3-(二甲基胺基)丙烯醯基)-3'-氟-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68c(600 mg, 1.1 mmol)、2-羥基-2-甲基丙脒(561 mg, 5.5 mmol)以及碳酸鉀(607 mg, 4.4 mmol)中加入 N, N-二甲基甲醯胺(4 mL)，反應液在55攝氏度下攪拌過夜，反應結束後，飽和食鹽水洗，乙酸乙酯萃取，有機相合併後乾燥濃縮並經矽膠管柱(乙酸乙酯)分離純化得到粗品，再經製備級管柱純化得到3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68(160 mg)，收率25%。 N,N-dimethylformamide (4 mmol) was added to ( E )-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5', 6 -dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-68c (600 mg, 1.1 mmol), 2-hydroxy- 2 -methylpropionamidine (561 mg, 5.5 mmol) and potassium carbonate (607 mg, 4.4 mmol). mL), the reaction solution was stirred at 55 degrees Celsius overnight. After the reaction was completed, it was washed with saturated brine and extracted with ethyl acetate. The organic phases were combined, dried, concentrated, and separated and purified by a silica gel column (ethyl acetate) to obtain a crude product, which was then purified by a preparative grade column to obtain 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-68 (160 mg) with a yield of 25%.

化合物Cpd-68經第一次掌性拆分得到光學純的化合物Cpd-68A(t _R= 1.215 min, 30 mg)和Cpd-68D(t _R= 2.663 min, 30 mg)，以及Cpd-68B和Cpd-68C的混合物Cpd-68M(t _R= 1.771 min)。 Compound Cpd-68 was subjected to the first chiral resolution to obtain optically pure compounds Cpd-68A (t _R = 1.215 min, 30 mg) and Cpd-68D (t _R = 2.663 min, 30 mg), as well as a mixture of Cpd-68B and Cpd-68C, Cpd-68M (t _R = 1.771 min).

Cpd-68M經第二次掌性拆分得到光學純的化合物Cpd-68B (t _R= 1.371 min, 22 mg)和Cpd-68C (t _R= 1.777 min, 24 mg)。 Cpd-68M was subjected to a second chiral resolution to afford optically pure compounds Cpd-68B ( t _R = 1.371 min, 22 mg) and Cpd-68C ( t _R = 1.777 min, 24 mg).

Cpd-68ACpd-68A

MS m/z(ESI)：590.1(M+1)。MS m/z(ESI):590.1(M+1).

HPLC：91.24 %(214 nm), 90.34%(254 nm)。HPLC: 91.24%(214 nm), 90.34%(254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.01(d, J= 5.2 Hz, 1H), 8.76(s, 1H), 8.58(d, J= 2.3 Hz, 1H), 8.09 – 8.03(m, 1H), 8.02(d, J= 5.2 Hz, 1H), 6.65(s, 1H), 6.06(q, J= 6.3 Hz, 1H), 5.05(s, 1H), 2.15(s, 3H), 1.98(s, 3H), 1.73(d, J= 6.4 Hz, 3H), 1.51(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.01(d, J = 5.2 Hz, 1H), 8.76(s, 1H), 8.58(d, J = 2.3 Hz, 1H), 8.09 – 8.03(m, 1H), 8.02(d, J = 5.2 Hz, 1H), 6.65(s, 1H), 6.06(q, J = 6.3 Hz, 1H), 5.05(s, 1H), 2.15(s, 3H), 1.98(s, 3H), 1.73(d, J = 6.4 Hz, 3H), 1.51(s, 6H).

Cpd-68BCpd-68B

MS m/z(ESI)：590.2(M+1)。MS m/z (ESI): 590.2 (M+1).

HPLC：99.65 %(214 nm), 99.77%(254 nm)。HPLC: 99.65% (214 nm), 99.77% (254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.01(d, J= 5.2 Hz, 1H), 8.74(s, 1H), 8.58(d, J= 2.3 Hz, 1H), 8.09 – 8.04(m, 1H), 8.02(d, J= 5.1 Hz, 1H), 6.67(s, 1H), 6.07(q, J= 6.2 Hz, 1H), 5.06(s, 1H), 2.11(s, 3H), 1.99(s, 3H), 1.73(d, J= 6.4 Hz, 3H), 1.52(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.01(d, J = 5.2 Hz, 1H), 8.74(s, 1H), 8.58(d, J = 2.3 Hz, 1H), 8.09 – 8.04(m, 1H), 8.02(d, J = 5.1 Hz, 1H), 6.67(s, 1H), 6.07(q, J = 6.2 Hz, 1H), 5.06(s, 1H), 2.11(s, 3H), 1.99(s, 3H), 1.73(d, J = 6.4 Hz, 3H), 1.52(s, 6H).

Cpd-68CCpd-68C

MS m/z(ESI)：590.2(M+1)。MS m/z (ESI): 590.2 (M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.01(d, J= 5.2 Hz, 1H), 8.75(s, 1H), 8.59(d, J= 2.3 Hz, 1H), 8.09 – 8.03(m, 1H), 8.02(d, J= 5.1 Hz, 1H), 6.68(s, 1H), 6.07(q, J= 6.3 Hz, 1H), 5.07(s, 1H), 2.12(s, 3H), 1.99(s, 3H), 1.73(d, J= 6.4 Hz, 3H), 1.52(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.01(d, J = 5.2 Hz, 1H), 8.75(s, 1H), 8.59(d, J = 2.3 Hz, 1H), 8.09 – 8.03(m, 1H), 8.02(d, J = 5.1 Hz, 1H), 6.68(s, 1H), 6.07(q, J = 6.3 Hz, 1H), 5.07(s, 1H), 2.12(s, 3H), 1.99(s, 3H), 1.73(d, J = 6.4 Hz, 3H), 1.52(s, 6H).

Cpd-68DCpd-68D

MS m/z(ESI)：590.2(M+1)。MS m/z (ESI): 590.2 (M+1).

HPLC：99.52 %(214 nm), 99.56%(254 nm)。HPLC: 99.52% (214 nm), 99.56% (254 nm).

¹H NMR(400 MHz, DMSO- d ₆)δ 9.00(d, J= 5.2 Hz, 1H), 8.75(s, 1H), 8.58(d, J= 2.1 Hz, 1H), 8.05(dd, J= 14.2, 5.6 Hz, 1H), 8.01(d, J= 5.3 Hz, 1H), 6.65(s, 1H), 6.06(q, J= 6.2 Hz, 1H), 5.05(s, 1H), 2.15(s, 3H), 1.98(s, 3H), 1.73(d, J= 6.3 Hz, 3H), 1.50(s, 6H). ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.00(d, J = 5.2 Hz, 1H), 8.75(s, 1H), 8.58(d, J = 2.1 Hz, 1H), 8.05(dd, J = 14.2, 5.6 Hz, 1H), 8.01(d, J = 5.3 Hz, 1H), 6.65(s, 1H), 6.06(q, J = 6.2 Hz, 1H), 5.05(s, 1H), 2.15(s, 3H), 1.98(s, 3H), 1.73(d, J = 6.3 Hz, 3H), 1.50(s, 6H).

第一次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK AD_3, 3*150mm, 3μm；流動相：CO ₂/MeOH(0.1% DEA)=80/20。 Conditions for the first separation: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALPAK AD_3, 3*150mm, 3μm; Mobile phase: CO ₂ /MeOH(0.1% DEA)=80/20.

第二次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：CHIRALPAK OD_3, 3*150mm, 3μm；流動相：CO ₂/MeOH(0.1% DEA)=80/20。 Second splitting conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: CHIRALPAK OD_3, 3*150mm, 3μm; Mobile phase: CO ₂ /MeOH(0.1% DEA)=80/20.

實施例15 Embodiment 15

第一步first step

2’-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮的製備 Preparation of 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2’-氯-4-羥基-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-01c(900 mg，3 mmol)、2-(1-氯乙基)-3,5-二氟吡啶 Cpd-67f(699 mg，3 mol)溶於N,N-二甲基甲醯胺(20 mL)，隨後加入18-冠醚-6(94 mg，0.3 mol)以及碳酸鉀(992 mg，7 mol)，反應液在60攝氏度下攪拌1小時，反應結束後，將反應液倒入冰水(200ml)中，用乙酸乙酯萃取(3 × 100 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)純化後得到2’-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-89a(940 mg)，產率：60%。 2'-Chloro-4-hydroxy-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-01c (900 mg, 3 mmol) and 2-(1-chloroethyl)-3,5-difluoropyridine Cpd-67f (699 mg, 3 mol) were dissolved in N,N-dimethylformamide (20 mL), and then 18-crown-6 (94 mg, 0.3 mol) and potassium carbonate (992 mg, 7 mol) were added. The reaction solution was stirred at 60 degrees Celsius for 1 hour. After the reaction was completed, the reaction solution was poured into ice water (200 ml) and extracted with ethyl acetate (3 × 100 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-89a (940 mg) in a yield of 60%.

MS m/z(ESI)：392.0(M+1)。MS m/z(ESI):392.0(M+1).

第二步Step 2

2',3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮的製備 Preparation of 2',3-dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

向2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-89a(940 mg，2 mmol)、N-氯代丁二醯亞胺(352 mg，2 mmol)、二氯乙酸(30 mg，0.2 mmol)加入異丙醇(20 mL)，反應在60攝氏度下反應3小時，反應結束後，加入水(200 mL)淬滅，用乙酸乙酯萃取(3 × 100 mL)。有機相用無水硫酸鈉乾燥、過濾，濾液濃縮後經矽膠管柱層析純化(流動相：石油醚/乙酸乙酯=2/1)純化後得到2’,3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-89b(780 mg)，產率：68.6%。 Isopropanol (20 mL) was added to 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-89a (940 mg, 2 mmol), N-chlorosuccinimide (352 mg, 2 mmol) and dichloroacetic acid (30 mg, 0.2 mmol). The reaction was carried out at 60 °C for 3 hours. After the reaction was completed, water (200 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (3 × 100 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 2',3-dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-89b (780 mg) with a yield of 68.6%.

MS m/z(ESI)：426.1(M+1)。MS m/z(ESI):426.1(M+1).

¹H NMR(400 MHz, DMSO- d6)δ 8.59(d, J= 2.3 Hz, 1H), 8.52(s, 1H), 8.05(dd, J= 3.8, 1.6 Hz, 1H), 7.67(s, 1H), 6.59(s, 2H), 2.73(s, 1H), 2.56(s, 9H). ¹ H NMR (400 MHz, DMSO- d6 )δ 8.59(d, J = 2.3 Hz, 1H), 8.52(s, 1H), 8.05(dd, J = 3.8, 1.6 Hz, 1H), 7.67(s, 1H), 6.59(s, 2H), 2.73(s, 1H), 2.56(s, 9H).

第三步third step

3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2’-(2-氟-3-(2-羥基丙-2-基)苯基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮的製備 Preparation of 3-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(2-fluoro-3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one

將2’,3-二氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-89b(780 mg，1 mmol)、2-(2-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼雜環戊烷-2-基)苯基)丙-2-醇 Cpd-89c(市售)(563 mg，2 mmol)、碳酸鈉(387 mg，3 mmol)和1,1’-雙(二苯基膦基)二茂鐵二氯化鈀(211 mg，0.1 mmol)溶於四氫呋喃：水=2：1(15 mL)。反應液在90攝氏度下攪拌1小時，反應結束後，加入水(100 mL)淬滅，用乙酸乙酯萃取(3 × 50 mL)，有機相用無水硫酸鈉乾燥、過濾，濾液進行濃縮。濃縮液通過製備純化凍乾得到3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2’-(2-氟-3-(2-羥基丙-2-基)苯基)-5’,6-二甲基-2 H-[1,4’-聯吡啶]-2-酮 Cpd-89(113.2 mg)，產率：11.4%。 2',3-Dichloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-89b (780 mg, 1 mmol), 2-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol Cpd-89c (commercially available) (563 mg, 2 mmol), sodium carbonate (387 mg, 3 mmol) and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (211 mg, 0.1 mmol) were dissolved in tetrahydrofuran:water = 2:1 (15 mL). The reaction solution was stirred at 90 degrees Celsius for 1 hour. After the reaction was completed, water (100 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was purified by preparative freeze drying to obtain 3-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(2-fluoro-3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-89 (113.2 mg) with a yield of 11.4%.

化合物Cpd-89經第一次掌性拆分得到化合物Cpd-89A和Cpd-89B的混合物Cpd-89M(t _R= 1.627 min，45 mg)，化合物Cpd-89C(t _R= 1.966 min, 25 mg)和Cpd-89D(t _R= 2.362 min, 37 mg)；混合物Cpd-89M經第二次掌性拆分得到光學純的化合物Cpd-89A(t _R= 3.043 min, 10 mg)和Cpd-89B (t _R= 3.258 min, 14 mg)。 The first chiral separation of compound Cpd-89 gave a mixture of compounds Cpd-89A and Cpd-89B, Cpd-89M (t _R = 1.627 min, 45 mg), compounds Cpd-89C (t _R = 1.966 min, 25 mg) and Cpd-89D (t _R = 2.362 min, 37 mg); the second chiral separation of the mixture Cpd-89M gave optically pure compounds Cpd-89A (t _R = 3.043 min, 10 mg) and Cpd-89B ( t _R = 3.258 min, 14 mg).

Cpd-89ACpd-89A

MS m/z(ESI)：544.2(M+1)。MS m/z(ESI):544.2(M+1).

HPLC：99.57%(214 nm), 100%(254 nm)。HPLC: 99.57%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.85(s, 1H), 8.39(d, J= 2.3 Hz, 1H), 7.98 – 7.93(m, 1H), 7.75(t, J= 7.5 Hz, 1H), 7.67(d, J= 1.3 Hz, 1H), 7.36 – 7.32(m, 1H), 7.32 – 7.28(m, 1H), 6.18(s, 1H), 5.78(dd, J= 13.1, 6.5 Hz, 1H), 2.18(s, 3H), 1.96(s, 3H), 1.86(d, J= 6.6 Hz, 3H), 1.68(d, J= 4.9 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.85(s, 1H), 8.39(d, J = 2.3 Hz, 1H), 7.98 – 7.93(m, 1H), 7.75(t, J = 7.5 Hz, 1H), 7.67(d, J = 1.3 Hz, 1H), 7.36 – 7.32(m, 1H), 7.32 – 7.28(m, 1H), 6.18(s, 1H), 5.78(dd, J = 13.1, 6.5 Hz, 1H), 2.18(s, 3H), 1.96(s, 3H), 1.86(d, J = 6.6 Hz, 3H), 1.68(d, J = 4.9 Hz, 6H).

Cpd-89BCpd-89B

MS m/z(ESI)：544.2(M+1)。MS m/z(ESI):544.2(M+1).

HPLC：100%(214 nm), 100%(254 nm)。HPLC: 100%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.91(s, 1H), 8.39(d, J= 2.2 Hz, 1H), 7.98(t, J= 7.0 Hz, 1H), 7.77(t, J= 7.1 Hz, 1H), 7.70(d, J= 0.8 Hz, 1H), 7.36(t, J= 7.8 Hz, 1H), 7.31 – 7.28(m, 1H), 6.18(s, 1H), 5.79(dd, J= 13.2, 6.6 Hz, 1H), 2.25(s, 3H), 1.95(s, 3H), 1.87(d, J= 6.6 Hz, 3H), 1.66(d, J= 3.7 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.91(s, 1H), 8.39(d, J = 2.2 Hz, 1H), 7.98(t, J = 7.0 Hz, 1H), 7.77(t, J = 7.1 Hz, 1H), 7.70(d, J = 0.8 Hz, 1H), 7.36(t, J = 7.8 Hz, 1H), 7.31 – 7.28(m, 1H), 6.18(s, 1H), 5.79(dd, J = 13.2, 6.6 Hz, 1H), 2.25(s, 3H), 1.95(s, 3H), 1.87(d, J = 6.6 Hz, 3H), 1.66(d, J = 3.7 Hz, 6H).

Cpd-89CCpd-89C

MS m/z(ESI)：544.2(M+1)。MS m/z(ESI):544.2(M+1).

HPLC：98.91%(214 nm), 99.42%(254 nm)。HPLC: 98.91%(214 nm), 99.42%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.82(s, 1H), 8.39(d, J= 2.2 Hz, 1H), 7.94(t, J= 6.8 Hz, 1H), 7.72(t, J= 7.3 Hz, 1H), 7.64(s, 1H), 7.30(ddd, J= 6.7, 5.8, 5.2 Hz, 2H), 6.16(s, 1H), 5.78(d, J= 6.6 Hz, 1H), 2.16(s, 3H), 1.96(s, 3H), 1.86(d, J= 6.5 Hz, 3H), 1.67(d, J= 4.7 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.82(s, 1H), 8.39(d, J = 2.2 Hz, 1H), 7.94(t, J = 6.8 Hz, 1H), 7.72(t, J = 7.3 Hz, 1H), 7.64(s, 1H), 7.30(ddd, J = 6.7, 5.8, 5.2 Hz, 2H), 6.16(s, 1H), 5.78(d, J = 6.6 Hz, 1H), 2.16(s, 3H), 1.96(s, 3H), 1.86(d, J = 6.5 Hz, 3H), 1.67(d, J = 4.7 Hz, 6H).

Cpd-89DCpd-89D

MS m/z(ESI)：544.2(M+1)。MS m/z(ESI):544.2(M+1).

HPLC：99.53%(214 nm), 100%(254 nm)。HPLC: 99.53%(214 nm), 100%(254 nm).

¹H NMR(400 MHz, CDCl ₃)δ 8.76(s, 1H), 8.38(d, J= 2.3 Hz, 1H), 7.87(td, J= 7.5, 1.6 Hz, 1H), 7.63(td, J= 7.7, 1.6 Hz, 1H), 7.51(d, J= 1.9 Hz, 1H), 7.29(dd, J= 7.9, 2.2 Hz, 1H), 7.24(d, J= 7.9 Hz, 1H), 6.12(s, 1H), 5.78(q, J= 6.5 Hz, 1H), 2.16(s, 3H), 1.94(s, 3H), 1.86(d, J= 6.5 Hz, 3H), 1.65(d, J= 2.8 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.76(s, 1H), 8.38(d, J = 2.3 Hz, 1H), 7.87(td, J = 7.5, 1.6 Hz, 1H), 7.63(td, J = 7.7, 1.6 Hz, 1H), 7.51(d, J = 1.9 Hz, 1H), 7.29(dd, J = 7.9, 2.2 Hz, 1H), 7.24(d, J = 7.9 Hz, 1H), 6.12(s, 1H), 5.78(q, J = 6.5 Hz, 1H), 2.16(s, 3H), 1.94(s, 3H), 1.86(d, J = 6.5 Hz, 3H), 1.65(d, J = 2.8 Hz, 6H).

第一次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號： Daicel CHIRALCEL OJ, 250mm 30 mm I.D.，10μm；流動相：CO ₂/MeOH[0.2%NH ₃(7M Solution in MeOH)]= 85/15。 Conditions for the first separation: Instrument brand: Waters Acquity UPCC; Preparation-grade column model: Daicel CHIRALCEL OJ, 250mm 30 mm ID, 10μm; Mobile phase: CO ₂ /MeOH [0.2% NH ₃ (7M Solution in MeOH)] = 85/15.

第二次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALCEL AD, 250mm 30 mm I.D.，10μm 流動相：CO ₂/MeOH[0.2%NH ₃(7M Solution in MeOH)]= 70/30。 Second splitting conditions: Instrument brand: Waters Acquity UPCC; Preparation grade column model: Daicel CHIRALCEL AD, 250mm 30 mm ID, 10μm; Mobile phase: CO ₂ /MeOH [0.2% NH ₃ (7M Solution in MeOH)] = 70/30.

實施例16 Embodiment 16

第一步first step

將3-溴-2'-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2H-[1,4'-聯吡啶] -2-酮Cpd-67b(200mg, 0.4093mmol)溶於1,4-二氧六環中(10 mL)，加入2-乙醯基-6-三甲基錫基吡啶(140mg, 0.5 mmol)、雙三苯基膦二氯化鈀(22.76 mg, 0.0324 mmol)。於80度封管反應2小時，反應結束後，過濾富集，殘液經製備級管柱(乙腈/水=65/35)純化得到6''-乙醯基-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4':2',2''-三聯吡啶]-2-酮Cpd-91a(93.8 mg)。產率：40%。 3-Bromo-2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one Cpd-67b (200 mg, 0.4093 mmol) was dissolved in 1,4-dioxane (10 mL), and 2-acetyl-6-trimethyltinylpyridine (140 mg, 0.5 mmol) and bistriphenylphosphinepalladium dichloride (22.76 mg, 0.0324 mmol) were added. The reaction was carried out at 80°C for 2 hours. After the reaction was completed, the residue was filtered and concentrated, and the residue was purified by a preparative column (acetonitrile/water = 65/35) to obtain 6''-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4':2',2''-terpyridine]-2-one Cpd-91a (93.8 mg). Yield: 40%.

MS m/z(ESI)：573.07(M+1)。MS m/z(ESI):573.07(M+1).

第二步Step 2

將6''-乙醯基-3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-5',6-二甲基-2 H-[1,4':2',2''-三聯吡啶]-2-酮Cpd91a(50 mg, 0.087mmol)溶於二氧六環(20 mL)中，加入甲基硼酸(8.0mg, 0.1305 mol)、碳酸銫(85mg, 0.3mmol)、Pd(dppf)Cl ₂(6mg, 0.008mmol)，於90度反應16小時，反應結束後。加水，乙酸乙酯萃取。收集有機相，乾燥過濾富集。殘液經矽膠管柱(石油醚/乙酸乙酯=3/1)得到6''-乙醯基-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-3,5',6-三甲基-2 H-[1,4':2',2''-三聯吡啶]-2-酮Cpd-91b(42mg)，產率：95%。 Dissolve 6''-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl- 2H- [1,4':2',2''-terpyridine]-2-one Cpd91a (50 mg, 0.087mmol) in dioxane (20 mL), add methylboric acid (8.0mg, 0.1305 mol), cesium carbonate (85mg, 0.3mmol), Pd(dppf)Cl ₂ (6mg, 0.008mmol), react at 90 degrees for 16 hours, and after the reaction is completed, add water and extract with ethyl acetate. Collect the organic phase, dry and filter to enrich. The residue was purified by silica gel column (petroleum ether/ethyl acetate = 3/1) to give 6''-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-3,5',6-trimethyl- 2H- [1,4':2',2''-terpyridine]-2-one Cpd-91b (42 mg) in a yield of 95%.

MS m/z(ESI)：509.1(M+1)。MS m/z(ESI):509.1(M+1).

第三步third step

將6''-乙醯基-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-3,5',6-三甲基-2 H-[1,4':2',2''-三聯吡啶]-2-酮Cpd-91b (42 mg, 0.082mmol)溶於四氫呋喃(10 mL)中，加入甲基溴化鎂(20mg, 0.164mmol)，於0度反應3小時。反應結束後，加氯化銨飽和溶液，再加乙酸乙酯萃取，收集有機相，乾燥過濾富集。殘液經製備級管柱(乙腈/水=65%:35%)得到4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-6''-(2-羥基丙-2-基)-3,5',6-三甲基-2 H-[1,4':2',2''-三聯吡啶]-2-酮Cpd-91(40mg)，產率：93.0 %。 6''-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-3,5',6-trimethyl- 2H- [1,4':2',2''-terpyridine]-2-one Cpd-91b ( 42 mg, 0.082mmol) was dissolved in tetrahydrofuran (10 mL), methylmagnesium bromide (20mg, 0.164mmol) was added, and the mixture was reacted at 0 degrees for 3 hours. After the reaction was completed, a saturated ammonium chloride solution was added, and then ethyl acetate was added for extraction, and the organic phase was collected, dried, filtered and enriched. The residue was purified by a preparative column (acetonitrile/water = 65%:35%) to give 4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-6''-(2-hydroxypropan-2-yl)-3,5',6-trimethyl- 2H- [1,4':2',2''-terpyridine]-2-one Cpd-91 (40 mg) with a yield of 93.0%.

MS m/z(ESI)：525.1(M+1)。MS m/z(ESI):525.1(M+1).

化合物Cpd-91經掌性拆分(流動相：CO ₂/MeOH(0.1%DEA)= 70/30)得到Cpd-91A(t _R= 1.223 min)和Cpd-91B(t _R= 1.305 min)的混合物Cpd-91M(19mg)，Cpd-91C(t _R= 1.721 min，5mg)，Cpd-91D(t _R= 1.966 min，6mg)；混合物Cpd-91M再經第二次拆分(流動相：CO ₂/MeOH(0.1%DEA)= 75/25)得到Cpd-91A(t _R= 2.725 min, 8mg)，Cpd-91B(t _R= 2.190 min, 5mg)。 Compound Cpd-91 was subjected to chiral separation (mobile phase: CO ₂ /MeOH (0.1% DEA) = 70/30) to obtain a mixture of Cpd-91A (t _R = 1.223 min) and Cpd-91B (t _R = 1.305 min), Cpd-91M (19 mg), Cpd-91C (t _R = 1.721 min, 5 mg), and Cpd-91D (t _R = 1.966 min, 6 mg); the mixture Cpd-91M was further subjected to a second separation (mobile phase: CO ₂ /MeOH (0.1% DEA) = 75/25) to obtain Cpd-91A (t _R = 2.725 min, 8 mg), and Cpd-91B (t _R = 2.190 min, 5 mg).

Cpd-91ACpd-91A

MS m/z(ESI)：525.1(M+1)。MS m/z(ESI):525.1(M+1).

HPLC：98.99%(214 nm), 97.31%(254 nm)。HPLC: 98.99%(214 nm), 97.31%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.56(s, 1H), 8.37(d, J= 2.3 Hz, 1H), 8.06(d, J= 7.6 Hz, 1H), 7.93(t, J= 7.6 Hz, 1H), 7.43(d, J= 7.8 Hz, 1H), 6.10(s, 1H),5.71(q, J= 6.6 Hz, 1H), 2.19(s, 3H), 2.05(s, 3H), 1.96(s, 3H), 1.80(d, J= 6.5 Hz, 3H), 1.59(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56(s, 1H), 8.37(d, J = 2.3 Hz, 1H), 8.06(d, J = 7.6 Hz, 1H), 7.93(t, J = 7.6 Hz, 1H), 7.43(d, J = 7.8 Hz, 1H), 6.10(s, 1H),5.71(q, J = 6.6 Hz, 1H), 2.19(s, 3H), 2.05(s, 3H), 1.96(s, 3H), 1.80(d, J = 6.5 Hz, 3H), 1.59(s, 6H).

Cpd-91BCpd-91B

MS m/z(ESI)：525.1(M+1)。MS m/z(ESI):525.1(M+1).

HPLC：99.35%(214 nm), 98.86%(254 nm)。HPLC: 99.35%(214 nm), 98.86%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.53(s, 1H), 8.38(d, J= 2.3 Hz, 1H), 8.04(d, J= 7.7 Hz, 1H), 7.89(t, J= 7.9 Hz, 1H), 7.39(t, J= 13.3 Hz, 1H), 6.08(s, 1H),5.71(q, J= 6.6 Hz, 1H), 2.14(s, 3H), 2.05(s, 2H), 1.96(s, 3H), 1.79(d, J= 6.5 Hz, 3H), 1.58(d, J= 6.5 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.53(s, 1H), 8.38(d, J = 2.3 Hz, 1H), 8.04(d, J = 7.7 Hz, 1H), 7.89(t, J = 7.9 Hz, 1H), 7.39(t, J = 13.3 Hz, 1H), 6.08(s, 1H),5.71(q, J = 6.6 Hz, 1H), 2.14(s, 3H), 2.05(s, 2H), 1.96(s, 3H), 1.79(d, J = 6.5 Hz, 3H), 1.58(d, J = 6.5 Hz, 6H).

Cpd-91CCpd-91C

MS m/z(ESI)：525.1(M+1)。MS m/z(ESI):525.1(M+1).

HPLC：99.35%(214 nm), 98.33%(254 nm)。HPLC: 99.35%(214 nm), 98.33%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.66(s, 1H), 8.38(d, J= 2.3 Hz, 1H), 8.14(t, J= 8.7 Hz, 1H), 7.58(d, J= 7.3 Hz, 1H), 7.30 – 7.27(m, 1H), 6.11(s, 1H), 5.71(q, J= 6.6 Hz, 1H), 2.22(s, 3H), 2.05(s, 3H), 1.99(s, 3H), 1.80(d, J= 6.5 Hz, 3H), 1.72(s, 3H), 1.66(s, 3H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.66(s, 1H), 8.38(d, J = 2.3 Hz, 1H), 8.14(t, J = 8.7 Hz, 1H), 7.58(d, J = 7.3 Hz, 1H), 7.30 – 7.27(m, 1H), 6.11(s, 1H), 5.71(q, J = 6.6 Hz, 1H), 2.22(s, 3H), 2.05(s, 3H), 1.99(s, 3H), 1.80(d, J = 6.5 Hz, 3H), 1.72(s, 3H), 1.66(s, 3H).

Cpd-91DCpd-91D

MS m/z(ESI)：525.1(M+1)。MS m/z(ESI):525.1(M+1).

HPLC：99.72%(214 nm), 99.61%(254 nm)。HPLC: 99.72%(214 nm), 99.61%(254 nm).

¹H NMR(400 MHz, CDCl ₃) δ8.55(s, 1H), 8.38(d, J= 2.2 Hz, 1H), 8.06(d, J= 7.7 Hz, 1H), 7.93(t, J= 7.7 Hz, 1H), 7.44(d, J= 7.9 Hz, 1H), 6.09(s, 1H),5.71(q, J= 6.5 Hz, 1H), 2.15(s, 3H), 2.05(s, 3H), 1.97(s, 3H), 1.79(d, J= 6.5 Hz, 3H), 1.60(s, 6H). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.55(s, 1H), 8.38(d, J = 2.2 Hz, 1H), 8.06(d, J = 7.7 Hz, 1H), 7.93(t, J = 7.7 Hz, 1H), 7.44(d, J = 7.9 Hz, 1H), 6.09(s, 1H),5.71(q, J = 6.5 Hz, 1H), 2.15(s, 3H), 2.05(s, 3H), 1.97(s, 3H), 1.79(d, J = 6.5 Hz, 3H), 1.60(s, 6H).

第一次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK OX_3, 3*150mm, 3μm；流動相：A/B：CO ₂/MeOH(0.1%DEA)=70/30。 Conditions for the first separation: Instrument brand: Waters Acquity UPCC; Preparation-grade column model: Daicel CHIRALPAK OX_3, 3*150mm, 3μm; Mobile phase: A/B: CO ₂ /MeOH(0.1%DEA)=70/30.

第二次拆分條件：儀器品牌：Waters Acquity UPCC；製備級管柱型號：Daicel CHIRALPAK IC_3, 3.0*150mm, 3μm；流動相：A/B：CO ₂/MeOH(0.1%DEA)=75/25。 Second splitting conditions: Instrument brand: Waters Acquity UPCC; Preparation column model: Daicel CHIRALPAK IC_3, 3.0*150mm, 3μm; Mobile phase: A/B: CO ₂ /MeOH(0.1%DEA)=75/25.

實施例17 Embodiment 17

將3-氯-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2'-(2-氟-3-(2-羥基丙-2-基)苯基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-89D(50 mg, 0.092mmol)溶於二氧六環(20 mL)中，加入甲基硼酸(8.0mg, 0.1305 mol)、碳酸銫(85mg, 0.3mmol)、Pd(dppf)Cl ₂(6mg, 0.008mmol)，於100度反應16小時，反應結束後。加水，乙酸乙酯萃取。收集有機相，乾燥過濾富集。殘液經矽膠管柱(石油醚/乙酸乙酯=3/1)純化得到4-(1-(3,5-二氟吡啶-2-基)乙氧基)-2'-(2-氟-3-(2-羥基丙烷-2-基)苯基)-3,5',6-三甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-92D(7.2mg)，產率：15%。 3-Chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(2-fluoro-3-(2-hydroxypropan-2-yl)phenyl)-5',6-dimethyl- 2H- [1,4'-bipyridinyl]-2-one Cpd-89D (50 mg, 0.092mmol) was dissolved in dioxane (20 mL), methylboric acid (8.0mg, 0.1305 mol), cesium carbonate (85mg, 0.3mmol), Pd(dppf)Cl ₂ (6mg, 0.008mmol) were added, and the reaction was carried out at 100 degrees for 16 hours. After the reaction was completed, water was added and extracted with ethyl acetate. The organic phase was collected, dried and filtered for enrichment. The residue was purified by silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(2-fluoro-3-(2-hydroxypropan-2-yl)phenyl)-3,5',6-trimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-92D (7.2 mg) in a yield of 15%.

MS m/z(ESI)：524.5(M+1)。MS m/z (ESI): 524.5 (M+1).

¹H NMR(400 MHz, CDCl ₃)δ 8.71(s, 1H), 8.37(d, J = 2.3 Hz, 1H), 7.84(td, J = 7.6, 1.8 Hz, 1H), 7.59(td, J = 7.7, 1.8 Hz, 1H), 7.48(d, J = 2.2 Hz, 1H), 7.28(d, J = 0.9 Hz, 1H), 7.22(d, J = 2.0 Hz, 1H), 6.05(s, 1H), 5.72 – 5.68(m, 1H), 2.13(s, 3H), 2.02(s, 3H), 1.91(s, 3H), 1.80(d, J = 6.5 Hz, 3H), 1.65(d, J = 3.0 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.71(s, 1H), 8.37(d, J = 2.3 Hz, 1H), 7.84(td, J = 7.6, 1.8 Hz, 1H), 7.59(td, J = 7.7, 1.8 Hz, 1H), 7.48(d, J = 2.2 Hz, 1H), 7.28(d, J = 0.9 Hz, 1H), 7.22(d, J = 2.0 Hz, 1H), 6.05(s, 1H), 5.72 – 5.68(m, 1H), 2.13(s, 3H), 2.02(s, 3H), 1.91(s, 3H), 1.80(d, J = 6.5 Hz, 3H), 1.65(d, J = 3.0 Hz, 6H).

實施例18 Embodiment 18

將3-溴-4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-5',6-二甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-68A(50 mg, 0.085mmol)溶於二氧六環(20 mL)中，加入甲基硼酸(8.0mg, 0.1305 mol)、碳酸銫(85mg, 0.3mmol)、Pd(dppf)Cl ₂(6mg, 0.008mmol)，於90度反應16小時，反應結束後，加水，乙酸乙酯萃取，收集有機相，乾燥過濾富集。殘液經矽膠管柱(石油醚/乙酸乙酯=3/1)純化得到4-(1-(3,5-二氟吡啶-2-基)乙氧基)-3'-氟-2'-(2-(2-羥基丙烷-2-基)嘧啶-4-基)-3,5',6-三甲基-2 H-[1,4'-聯吡啶]-2-酮Cpd-93A(35.7mg)，產率：80%。 3-Bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5',6-dimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-68A (50 mg, 0.085 mmol) was dissolved in dioxane (20 mL), and methylboric acid (8.0 mg, 0.1305 mol), cesium carbonate (85 mg, 0.3 mmol), and Pd(dppf)Cl ₂ (6 mg, 0.008 mmol) were added. The mixture was reacted at 90°C for 16 hours. After the reaction was completed, water was added, and the mixture was extracted with ethyl acetate. The organic phase was collected, dried, filtered, and enriched. The residue was purified by silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-3,5',6-trimethyl- 2H- [1,4'-bipyridyl]-2-one Cpd-93A (35.7 mg) in a yield of 80%.

MS m/z(ESI)：526.2(M+1)。MS m/z(ESI):526.2(M+1).

¹H NMR(400 MHz, CDCl ₃)δ 8.87(d, J = 5.1 Hz, 1H), 8.57(s, 1H), 8.37(d, J = 2.1 Hz, 1H), 8.00(d, J = 5.1 Hz, 1H), 7.29(d, J = 2.2 Hz, 1H), 6.12(s, 1H), 5.71(q, J = 6.4 Hz, 1H), 2.21(s, 3H), 2.05(s, 3H), 1.94(s, 3H), 1.81(d, J = 6.5 Hz, 3H), 1.60(d, J = 5.9 Hz, 6H). ¹ H NMR (400 MHz, CDCl ₃ )δ 8.87(d, J = 5.1 Hz, 1H), 8.57(s, 1H), 8.37(d, J = 2.1 Hz, 1H), 8.00(d, J = 5.1 Hz, 1H), 7.29(d, J = 2.2 Hz, 1H), 6.12(s, 1H), 5.71(q, J = 6.4 Hz, 1H), 2.21(s, 3H), 2.05(s, 3H), 1.94(s, 3H), 1.81(d, J = 6.5 Hz, 3H), 1.60(d, J = 5.9 Hz, 6H).

使用與上述實施例中類似的條件，製備了如下表1中的化合物。將這些化合物的結構表徵資料一並列於表1。Using conditions similar to those in the above examples, the compounds listed in Table 1 were prepared. The structural characteristics of these compounds are listed in Table 1.

表1 化合物編號 ¹H NMR 數據 Cpd-16 ¹H NMR(400 MHz, DMSO- d ₆)δ 8.96(d, J= 5.2 Hz, 1H), 8.87(s, 1H), 8.61(d, J= 2.4 Hz, 1H), 8.40(s, 1H), 8.23(d, J= 5.2 Hz, 1H), 8.10(td, J= 9.9, 2.4 Hz,1H), 6.98(s, 2H), 6.84(s, 1H), 5.49(s, 2H), 2.68(t, J= 7.8 Hz, 4H), 2.10(s, 3H), 1.97(s, 3H), 1.90 -1.78(m, 2H). Cpd-16A* ¹H NMR(400 MHz, DMSO- d ₆)δ 8.96(d, J= 5.2Hz, 1H), 8.87(s, 1H), 8.61(d, J= 2.2 Hz, 1H),8.40(s, 1H), 8.23(d, J= 5.2 Hz, 1H), 8.13 –8.07(m, 1H), 6.97(s, 2H), 6.84(s, 1H), 5.49(s, 2H), 2.68(t, J= 8.0 Hz, 4H), 2.10(s, 3H), 1.97(s, 3H), 1.83(dt, J= 15.6, 7.8 Hz, 2H). Cpd-18 ¹H NMR(400 MHz, DMSO- d ₆)δ 8.96(d, J= 5.2Hz, 1H), 8.87(s, 1H), 8.61(d, J= 2.2 Hz, 1H),8.40(s, 1H), 8.23(d, J= 5.2 Hz, 1H), 8.10(td, J= 9.8, 2.4 Hz, 1H), 6.97(s, 2H), 6.84(s, 1H),5.49(s, 2H), 2.68(t, J= 8.0 Hz, 4H), 2.10(s,3H), 1.97(s, 3H), 1.84(td, J= 10.8, 3.4 Hz, 2H). Cpd-31A* ¹H NMR(400 MHz, CDCl ₃)δ 8.57(s, 1H), 8.40(d, J = 1.8 Hz, 1H), 8.06(s, 1H), 7.90(d, J = 7.6 Hz, 1H), 7.59(d, J = 7.8 Hz, 1H), 7.54 – 7.47(m, 1H), 7.36 – 7.29(m, 1H), 6.44(s, 1H), 5.42(d, J = 1.5 Hz, 2H), 2.19(s, 3H), 2.04(s, 3H), 1.78(s, 6H) Cpd-42A* ¹H NMR(400 MHz, DMSO- d ₆)δ 8.72(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.09(d, J = 5.6 Hz, 2H), 7.91(d, J = 7.6 Hz, 1H), 7.68-7.61(m,, 2H), 6.89(s, 1H), 2.13(s, 3H), 2.07(s, 3H), 1.75(s, 6H). Cpd-43A* ¹H NMR(400 MHz, CDCl ₃)δ 8.55(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 7.99(s, 1H), 7.82(d, J = 6.2 Hz, 1H), 7.68 – 7.63(m, 1H), 7.58 – 7.51(m, 1H), 7.35 – 7.29(m, 1H), 6.45(s, 1H), 5.42(d, J = 1.4 Hz, 2H), 5.33(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.63(s, 6H). Cpd-44A* ¹H NMR(400 MHz, DMSO- d ₆)δ 8.72(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.09(d, J = 5.6 Hz, 2H), 7.91(d, J = 7.6 Hz, 1H), 7.68-7.61(m,, 2H), 6.89(s, 1H), 2.13(s, 3H), 2.07(s, 3H), 1.75(s, 6H). Cpd-56A* ¹H NMR(400 MHz, CDCl ₃)δ 8.76(d, J = 5.4 Hz, 1H), 8.41(d, J = 2.3 Hz, 1H), 8.11(d, J = 8.0 Hz, 1H), 7.89(s, 1H), 7.60(d, J = 5.3 Hz, 1H), 7.52(d, J = 8.1 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.65(d, J = 3.5 Hz, 6H). Cpd-57A* ¹H NMR(400 MHz, CDCl ₃)δ 8.77(s, 1H), 8.41(d, J = 2.2 Hz, 1H), 8.11(d, J = 7.0 Hz, 1H), 7.91(s, 1H), 7.62(s, 1H), 7.53(d, J = 5.3 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 5.42(s, 2H), 2.17(s, 3H), 1.99(s, 3H), 1.66(s, 6H). Cpd-61 ¹H NMR(400 MHz, DMSO- d ₆)δ 8.77(s, 1H), 8.59(d, J = 2.3 Hz, 1H), 8.29(s, 1H), 8.26(d, J = 7.7 Hz, 1H), 8.09(td, J = 9.9, 2.3 Hz, 1H), 7.93(t, J = 7.8 Hz, 1H), 7.63(d, J = 7.8 Hz, 1H), 6.81(s, 1H), 5.47(s, 2H), 2.59(dtd, J = 18.2, 9.1, 4.6 Hz, 2H), 2.23(dd, J = 19.1, 9.5 Hz, 2H), 2.05(s, 3H), 1.97(s, 3H), 1.93 – 1.78(m, 2H). Cpd-62A* ¹H NMR(400 MHz, CDCl ₃)δ 8.41(d, J = 2.3 Hz, 1H), 7.99(d, J = 6.8 Hz, 1H), 7.82(dd, J = 16.6, 9.0 Hz, 2H), 7.65(d, J = 7.7 Hz, 1H), 7.48(d, J = 8.0 Hz, 1H), 7.37 – 7.29(m, 2H), 6.36(s, 1H), 2.14(s, 3H), 2.03(s, 3H), 1.60(d, J = 8.9 Hz, 6H). Cpd-65A* ¹H NMR(400 MHz, DMSO- d ₆)δ 9.02(d, J = 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J = 2.3 Hz, 1H), 8.10(td, J = 9.9, 2.3 Hz, 1H), 8.03(d, J = 5.2 Hz, 1H), 6.86(s, 1H), 5.51(s, 2H), 5.08(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H). Cpd-75A* ¹H NMR(400 MHz, CDCl ₃)δ 8.60(s, 1H), 8.40(d, J = 2.2 Hz, 1H), 7.71(t, J = 7.3 Hz, 1H), 7.55(s, 1H), 7.37 – 7.27(m, 2H), 6.44(s, 1H), 5.41(s, 2H), 2.23(s, 3H), 2.08(s, 3H), 1.66(d, J = 5.7 Hz, 6H) Cpd-78B* ¹H NMR(400 MHz, DMSO- d ₆)δ 8.73(s, 1H), 8.60(d, J= 2.4 Hz, 1H), 8.13 – 8.06(m, 1H), 7.75(dd, J= 7.0, 2.4 Hz, 1H), 7.66 – 7.61(m, 1H), 7.31 – 7.25(m, 1H), 6.89(s, 1H), 5.51(s, 2H), 2.14(s, 3H), 2.04(s, 3H), 1.46(s, 6H). Cpd-85A* ¹H NMR(400 MHz, CDCl ₃)δ 8.71(s, 1H), 8.40(d, J = 2.3 Hz, 1H), 8.14(s, 1H), 7.84(d, J = 7.8 Hz, 1H), 7.55(d, J = 9.5 Hz, 2H), 7.43(t, J = 7.7 Hz, 1H), 7.35 – 7.30(m, 1H), 6.40(s, 1H), 5.42(d, J = 1.7 Hz, 2H), 2.15(s, 3H), 1.99(s, 3H), 1.64(d, J = 1.2 Hz, 6H). Cpd-87A* ¹H NMR(400 MHz, CDCl ₃)δ 8.41(d, J = 2.3 Hz, 1H), 7.99(d, J = 6.8 Hz, 1H), 7.82(dd, J = 16.6, 9.0 Hz, 2H), 7.65(d, J = 7.7 Hz, 1H), 7.48(d, J = 8.0 Hz, 1H), 7.37 – 7.29(m, 2H), 6.36(s, 1H), 2.14(s, 3H), 2.03(s, 3H), 1.60(d, J = 8.9 Hz, 6H). Cpd-88A* ¹H NMR(400 MHz, CDCl ₃) δ8.56(s, 1H), 8.37(d, J= 2.2 Hz, 1H), 7.68(td, J= 7.8, 1.2 Hz, 1H), 7.51(dd, J= 10.2, 4.0 Hz, 1H), 7.30 – 7.24(m, 2H), 6.16(s, 1H), 5.78(q, J= 6.4 Hz, 1H), 2.19(s, 3H), 1.99(s, 3H), 1.86(d, J= 6.4 Hz, 3H), 1.63(d, J= 4.4 Hz, 6H). Cpd-95A* ¹H NMR(400 MHz, CDCl ₃) δ8.54(s, 1H), 8.36(d, J= 2.4 Hz, 1H), 7.66(td, J= 7.8, 1.6 Hz, 1H), 7.50(dd, J= 10.4, 4.0 Hz, 1H), 7.27-7.22(m, 2H), 6.09(s, 1H), 5.70(q, J= 6.6 Hz, 1H), 2.18(s, 3H), 2.04(s, 3H), 1.96(s, 3H), 1.80(d, J= 6.4 Hz, 3H), 1.64(d, J= 4.8 Hz, 6H). 註：*代表立體異構物 Table 1 Compound No. ¹ H NMR data Cpd-16 ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.96(d, J = 5.2 Hz, 1H), 8.87(s, 1H), 8.61(d, J = 2.4 Hz, 1H), 8.40(s, 1H), 8.23(d, J = 5.2 Hz, 1H), 8.10(td, J = 9.9, 2.4 Hz,1H), 6.98(s, 2H), 6.84(s, 1H), 5.49(s, 2H), 2.68(t, J = 7.8 Hz, 4H), 2.10(s, 3H), 1.97(s, 3H), 1.90 -1.78(m, 2H). Cpd-16A* ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.96(d, J = 5.2 Hz, 1H), 8.87(s, 1H), 8.61(d, J = 2.2 Hz, 1H),8.40(s, 1H), 8.23(d, J = 5.2 Hz, 1H), 8.13 –8.07(m, 1H), 6.97(s, 2H), 6.84(s, 1H), 5.49(s, 2H), 2.68(t, J = 8.0 Hz, 4H), 2.10(s, 3H), 1.97(s, 3H), 1.83(dt, J = 15.6, 7.8 Hz, 2H). Cpd-18 ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.96(d, J = 5.2 Hz, 1H), 8.87(s, 1H), 8.61(d, J = 2.2 Hz, 1H),8.40(s, 1H), 8.23(d, J = 5.2 Hz, 1H), 8.10(td, J = 9.8, 2.4 Hz, 1H), 6.97(s, 2H), 6.84(s, 1H),5.49(s, 2H), 2.68(t, J = 8.0 Hz, 4H), 2.10(s,3H), 1.97(s, 3H), 1.84(td, J = 10.8, 3.4 Hz, 2H). Cpd-31A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.57(s, 1H), 8.40(d, J = 1.8 Hz, 1H), 8.06(s, 1H), 7.90(d, J = 7.6 Hz, 1H), 7.59(d, J = 7.8 Hz, 1H), 7.54 – 7.47(m, 1H), 7.36 – 7.29(m, 1H), 6.44(s, 1H), 5.42(d, J = 1.5 Hz, 2H), 2.19(s, 3H), 2.04(s, 3H), 1.78(s, 6H) Cpd-42A* ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.72(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.09(d, J = 5.6 Hz, 2H), 7.91(d, J = 7.6 Hz, 1H), 7.68-7.61(m,, 2H), 6.89(s, 1H), 2.13(s, 3H), 2.07(s, 3H), 1.75(s, 6H). Cpd-43A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.55(s, 1H), 8.40(d, J = 2.1 Hz, 1H), 7.99(s, 1H), 7.82(d, J = 6.2 Hz, 1H), 7.68 – 7.63(m, 1H), 7.58 – 7.51(m, 1H), 7.35 – 7.29(m, 1H), 6.45(s, 1H), 5.42(d, J = 1.4 Hz, 2H), 5.33(s, 2H), 2.19(s, 3H), 2.05(s, 3H), 1.63(s, 6H). Cpd-44A* ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.72(s, 1H), 8.61(d, J = 2.2 Hz, 1H), 8.09(d, J = 5.6 Hz, 2H), 7.91(d, J = 7.6 Hz, 1H), 7.68-7.61(m,, 2H), 6.89(s, 1H), 2.13(s, 3H), 2.07(s, 3H), 1.75(s, 6H). Cpd-56A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.76(d, J = 5.4 Hz, 1H), 8.41(d, J = 2.3 Hz, 1H), 8.11(d, J = 8.0 Hz, 1H), 7.89(s, 1H), 7.60(d, J = 5.3 Hz, 1H), 7.52(d, J = 8.1 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 2.16(s, 3H), 1.99(s, 3H), 1.65(d, J = 3.5 Hz, 6H). Cpd-57A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.77(s, 1H), 8.41(d, J = 2.2 Hz, 1H), 8.11(d, J = 7.0 Hz, 1H), 7.91(s, 1H), 7.62(s, 1H), 7.53(d, J = 5.3 Hz, 1H), 7.35 – 7.30(m, 1H), 6.39(s, 1H), 5.42(s, 2H), 2.17(s, 3H), 1.99(s, 3H), 1.66(s, 6H). Cpd-61 ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.77(s, 1H), 8.59(d, J = 2.3 Hz, 1H), 8.29(s, 1H), 8.26(d, J = 7.7 Hz, 1H), 8.09(td, J = 9.9, 2.3 Hz, 1H), 7.93(t, J = 7.8 Hz, 1H), 7.63(d, J = 7.8 Hz, 1H), 6.81(s, 1H), 5.47(s, 2H), 2.59(dtd, J = 18.2, 9.1, 4.6 Hz, 2H), 2.23(dd, J = 19.1, 9.5 Hz, 2H), 2.05(s, 3H), 1.97(s, 3H), 1.93 – 1.78(m, 2H). Cpd-62A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.41(d, J = 2.3 Hz, 1H), 7.99(d, J = 6.8 Hz, 1H), 7.82(dd, J = 16.6, 9.0 Hz, 2H), 7.65(d, J = 7.7 Hz, 1H), 7.48(d, J = 8.0 Hz, 1H), 7.37 – 7.29(m, 2H), 6.36(s, 1H), 2.14(s, 3H), 2.03(s, 3H), 1.60(d, J = 8.9 Hz, 6H). Cpd-65A* ¹ H NMR (400 MHz, DMSO- d ₆ )δ 9.02(d, J = 5.2 Hz, 1H), 8.77(s, 1H), 8.60(d, J = 2.3 Hz, 1H), 8.10(td, J = 9.9, 2.3 Hz, 1H), 8.03(d, J = 5.2 Hz, 1H), 6.86(s, 1H), 5.51(s, 2H), 5.08(s, 1H), 2.16(s, 3H), 2.03(s, 3H), 1.52(s, 6H). Cpd-75A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.60(s, 1H), 8.40(d, J = 2.2 Hz, 1H), 7.71(t, J = 7.3 Hz, 1H), 7.55(s, 1H), 7.37 – 7.27(m, 2H), 6.44(s, 1H), 5.41(s, 2H), 2.23(s, 3H), 2.08(s, 3H), 1.66(d, J = 5.7 Hz, 6H) Cpd-78B* ¹ H NMR (400 MHz, DMSO- d ₆ )δ 8.73(s, 1H), 8.60(d, J = 2.4 Hz, 1H), 8.13 – 8.06(m, 1H), 7.75(dd, J = 7.0, 2.4 Hz, 1H), 7.66 – 7.61(m, 1H), 7.31 – 7.25(m, 1H), 6.89(s, 1H), 5.51(s, 2H), 2.14(s, 3H), 2.04(s, 3H), 1.46(s, 6H). Cpd-85A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.71(s, 1H), 8.40(d, J = 2.3 Hz, 1H), 8.14(s, 1H), 7.84(d, J = 7.8 Hz, 1H), 7.55(d, J = 9.5 Hz, 2H), 7.43(t, J = 7.7 Hz, 1H), 7.35 – 7.30(m, 1H), 6.40(s, 1H), 5.42(d, J = 1.7 Hz, 2H), 2.15(s, 3H), 1.99(s, 3H), 1.64(d, J = 1.2 Hz, 6H). Cpd-87A* ¹ H NMR (400 MHz, CDCl ₃ )δ 8.41(d, J = 2.3 Hz, 1H), 7.99(d, J = 6.8 Hz, 1H), 7.82(dd, J = 16.6, 9.0 Hz, 2H), 7.65(d, J = 7.7 Hz, 1H), 7.48(d, J = 8.0 Hz, 1H), 7.37 – 7.29(m, 2H), 6.36(s, 1H), 2.14(s, 3H), 2.03(s, 3H), 1.60(d, J = 8.9 Hz, 6H). Cpd-88A* ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56(s, 1H), 8.37(d, J = 2.2 Hz, 1H), 7.68(td, J = 7.8, 1.2 Hz, 1H), 7.51(dd, J = 10.2, 4.0 Hz, 1H), 7.30 – 7.24(m, 2H), 6.16(s, 1H), 5.78(q, J = 6.4 Hz, 1H), 2.19(s, 3H), 1.99(s, 3H), 1.86(d, J = 6.4 Hz, 3H), 1.63(d, J = 4.4 Hz, 6H). Cpd-95A* ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.54(s, 1H), 8.36(d, J = 2.4 Hz, 1H), 7.66(td, J = 7.8, 1.6 Hz, 1H), 7.50(dd, J = 10.4, 4.0 Hz, 1H), 7.27-7.22(m, 2H), 6.09(s, 1H), 5.70(q, J = 6.6 Hz, 1H), 2.18(s, 3H), 2.04(s, 3H), 1.96(s, 3H), 1.80(d, J = 6.4 Hz, 3H), 1.64(d, J = 4.8 Hz, 6H). Note: * represents stereoisomers

生物學評價Biological Evaluation

測試例 1 本發明化合物對重組人 p38 α -MK2 激酶抑制活性的測定1. 實驗目的：測定化合物的p38α-MK2激酶抑制IC50值。 2. 實驗材料： GST-MK2溶液濃度：15328.84 nM GST-P-p38α溶液濃度：3030 nM Hsp27多肽(FITC)濃度：1051.3 µM。 3. 實驗操作： (1)採用Echo進行化合物稀釋，最終濃度為10 µM ~ 0.17 nM，取200 nL溶液加入到檢測板孔中； (2)將5 µL 4倍稀釋的GST-MK2混合物加至檢測板孔中(含化合物)； (3)將15 µL 1.33倍稀釋的GST-P-p38a、ATP和Hsp27多肽混合物加至檢測板孔中(含化合物)； (4)1000轉/分，離心60秒左右，23℃溫育120分鐘； (5)加入60 µL IMAP溶液開始反應； (6)以1000轉/分離心檢測板約60秒，23℃溫育30分鐘； (7)在Neo2讀取檢測板(Ex/Em=485nm/FITC FP-P pol 528nm & FITC FP-S pol 528nm)； (8)通過訊號比值計算相對DMSO空白的相對酶活性抑制，利用軟體擬合曲線計算IC50值。 4. 實驗結果表2 本發明化合物對p38α-MK2激酶抑制活性測試結果 化合物編號 p38α-MK2 激酶抑制活性(nM) Cpd-01 18 Cpd-01A* 2.2 Cpd-02 8 Cpd-02B* 5.3 Cpd-16 14.4 Cpd-16A* 8.3 Cpd-17A* 12.6 Cpd-31A* 7.9 Cpd-37 1.6 Cpd-37B* 0.4 Cpd-40A* 0.7 Cpd-42A* 0.5 Cpd-43A* 1.3 Cpd-44A* 0.8 Cpd-45 9.3 Cpd-53A* 0.5 Cpd-54A* 0.9 Cpd-55 2.2 Cpd-55A* 1.1 Cpd-56A* 3.4 Cpd-57A* 3.6 Cpd-58 3 Cpd-61 2.9 Cpd-62A* 1.3 Cpd-65A* 0.98 Cpd-66A* 0.5 Cpd-67 1.2 Cpd-68A* 1.8 Cpd-75A* ＜0.1 Cpd-78B* 17.2 Cpd-81A* 1.9 Cpd-82A* 4.6 Cpd-85A* 5.3 Cpd-87A* 1.3 Cpd-88A* 0.7 Cpd-89D* 1.6 Cpd-91A* 1.8 Cpd-92D* 15.5 Cpd-93A* 8.4 Cpd-94A* 4.6 Cpd-95A* 1.6 註：*代表立體異構物 Test Example 1 Determination of the inhibitory activity of the compounds of the present invention on recombinant human p38α -MK2 kinase 1. Experimental purpose: Determine the p38α -MK2 kinase inhibition IC50 value of the compounds. 2. Experimental materials: GST-MK2 solution concentration: 15328.84 nM GST-P-p38α solution concentration: 3030 nM Hsp27 peptide (FITC) concentration: 1051.3 µM. 3. Experimental procedures: (1) Use Echo to dilute the compound to a final concentration of 10 µM ~ 0.17 nM, and add 200 nL of the solution to the wells of the assay plate; (2) Add 5 µL of a 4-fold diluted GST-MK2 mixture to the wells of the assay plate (containing the compound); (3) Add 15 µL of a 1.33-fold diluted GST-P-p38a, ATP, and Hsp27 peptide mixture to the wells of the assay plate (containing the compound); (4) Centrifuge at 1000 rpm for about 60 seconds and incubate at 23°C for 120 minutes; (5) Add 60 µL of IMAP solution to start the reaction; (6) Centrifuge the assay plate at 1000 rpm for about 60 seconds and incubate at 23°C for 30 minutes; (7) Read the assay plate on Neo2 (Ex/Em=485nm/FITC FP-P pol 528nm & FITC FP-S pol 528nm); (8) The relative enzyme activity inhibition relative to the DMSO blank was calculated by the signal ratio, and the IC50 value was calculated by using the software to fit the curve. 4. Experimental results Table 2 Test results of the inhibitory activity of the compounds of the present invention on p38α-MK2 kinase Compound No. p38α-MK2 kinase inhibitory activity (nM) Cpd-01 18 Cpd-01A* 2.2 Cpd-02 8 Cpd-02B* 5.3 Cpd-16 14.4 Cpd-16A* 8.3 Cpd-17A* 12.6 Cpd-31A* 7.9 Cpd-37 1.6 Cpd-37B* 0.4 Cpd-40A* 0.7 Cpd-42A* 0.5 Cpd-43A* 1.3 Cpd-44A* 0.8 Cpd-45 9.3 Cpd-53A* 0.5 Cpd-54A* 0.9 Cpd-55 2.2 Cpd-55A* 1.1 Cpd-56A* 3.4 Cpd-57A* 3.6 Cpd-58 3 Cpd-61 2.9 Cpd-62A* 1.3 Cpd-65A* 0.98 Cpd-66A* 0.5 Cpd-67 1.2 Cpd-68A* 1.8 Cpd-75A* <0.1 Cpd-78B* 17.2 Cpd-81A* 1.9 Cpd-82A* 4.6 Cpd-85A* 5.3 Cpd-87A* 1.3 Cpd-88A* 0.7 Cpd-89D* 1.6 Cpd-91A* 1.8 Cpd-92D* 15.5 Cpd-93A* 8.4 Cpd-94A* 4.6 Cpd-95A* 1.6 Note: * represents stereoisomers

實驗結果表明，本發明化合物具有良好的p38α-MK2激酶抑制活性。The experimental results show that the compound of the present invention has good p38α-MK2 kinase inhibitory activity.

測試例 2 本發明化合物對人外周血單核細胞 (PBMC) 中細胞因子 (TNF α、 IL-1 β、 IL-6) 的抑制活性測定1. 實驗目的：測定化合物對人外周血單核細胞(PBMC)中細胞因子(TNFα、IL-1β、IL-6)的抑制IC ₅₀值。 2. 實驗材料：人PBMC細胞、V-PLEX人TNF-α 檢測試劑盒 (Cat#K151QWD-2/MSD)。 3. 實驗步驟第一步：解凍PBMC(P121010902C, 1000萬/管)，並將培養基按照(190 μl/孔)轉移到96孔細胞培養板(200,000細胞/孔)，37℃，5% CO ₂孵育2h；第二步：化合物被等比例稀釋並滴加到測定板上(10 μl/孔)，37℃，5% CO ₂孵育1 h；第三步：用LPS(100 ng/ml)刺激PBMC，4 h(2 μL/孔)後離心，取上清(180 μl)，用MSD試劑盒測定細胞因子水準，上清保存於-80℃，用MSD試劑盒檢測；第四步：24小時後，用稀釋劑將PBMC樣品稀釋20倍檢測TNF-α，IL-1β和IL-6；第五步：加入50 μL/孔製備樣品到MSD板，用150 μL/孔的洗滌液洗滌3次；再加入25 μL/孔1X檢測抗體溶液；用150 μL/孔的洗滌液洗滌3次，添加150 μL/孔2X Read Buffer T，在MSD儀器上分析板。 4. 實驗結果表3 本發明化合物對人PBMC中細胞因子的抑制活性化合物編號人PBMC(TNFα)抑制活性 IC ₅₀₍nM) Cpd-01A* 29.4 Cpd-02A* 26.5 Cpd-16A* 13.8 Cpd-17B* 57.3 Cpd-31A* 4.4 Cpd-37 5.2 Cpd-40A* 7.8 Cpd-43A* 4.9 Cpd-53A* 8.7 Cpd-55A* 1.1 Cpd-56A* 13.2 Cpd-57A* 8.9 Cpd-61 4.0 Cpd-65A* 0.1 Cpd-67 0.2 Cpd-68A* 2.0 Cpd-78B* 2.7 Cpd-81A* 10.6 Cpd-87A* 2.0 Cpd-88A* 0.4 Cpd-89D* 2.3 Cpd-95A* 0.7 註：*代表立體異構物 Test Example 2 Determination of the inhibitory activity of the compounds of the present invention on cytokines (TNFα , IL-1β , IL-6) in human peripheral blood mononuclear cells ( PBMC ) 1. Experimental purpose: Determine the inhibitory _IC50 value of the compounds on cytokines (TNFα, IL-1β, IL-6) in human peripheral blood mononuclear cells (PBMC). 2. Experimental materials: human PBMC cells, V-PLEX human TNF-α detection kit (Cat#K151QWD-2/MSD). 3. Experimental procedures Step 1: thaw PBMC (P121010902C, 10 million/tube) and transfer the culture medium to a 96-well cell culture plate (200,000 cells/well) at (190 μl/well), incubate at 37°C, 5% CO ₂ for 2 h; Step 2: the compound was diluted in equal proportions and added dropwise to the assay plate (10 μl/well), incubate at 37°C, 5% CO ₂ for 1 h; Step 3: stimulate PBMC with LPS (100 ng/ml), centrifuge after 4 h (2 μL/well), take the supernatant (180 μl), and measure the cytokine level using the MSD kit. The supernatant was stored at -80°C and detected using the MSD kit; Step 4: After 24 hours, dilute the PBMC sample 20 times with diluent to detect TNF-α, IL-1β and IL-6; Step 5: Add 50 μL/well of the prepared sample to the MSD plate, wash 3 times with 150 μL/well of washing solution; then add 25 μL/well of 1X detection antibody solution; wash 3 times with 150 μL/well of washing solution, add 150 μL/well of 2X Read Buffer T, and analyze the plate on the MSD instrument. 4. Experimental results Table 3 Inhibitory activity of the compounds of the present invention on cytokines in human PBMC Compound No. Human PBMC (TNFα) inhibitory activity IC _{50 (} nM) Cpd-01A* 29.4 Cpd-02A* 26.5 Cpd-16A* 13.8 Cpd-17B* 57.3 Cpd-31A* 4.4 Cpd-37 5.2 Cpd-40A* 7.8 Cpd-43A* 4.9 Cpd-53A* 8.7 Cpd-55A* 1.1 Cpd-56A* 13.2 Cpd-57A* 8.9 Cpd-61 4.0 Cpd-65A* 0.1 Cpd-67 0.2 Cpd-68A* 2.0 Cpd-78B* 2.7 Cpd-81A* 10.6 Cpd-87A* 2.0 Cpd-88A* 0.4 Cpd-89D* 2.3 Cpd-95A* 0.7 Note: * represents stereoisomers

結果表明，本發明化合物對p38α-MK2通路下游的TNFα的炎症因子具有顯著的抑制作用。The results show that the compound of the present invention has a significant inhibitory effect on the inflammatory factor TNFα downstream of the p38α-MK2 pathway.

測試例 3 本發明代表性化合物對重組人 p38 α -PRAK 激酶抑制活性的測定及選擇性研究1. 實驗目的：測定化合物的p38α-PRAK激酶抑制IC50值，以評價化合物的選擇性。 2. 實驗材料 GST-PRAK溶液濃度：6710.15 nM GST-P-p38α溶液濃度：3030 nM Hsp27多肽(FITC)濃度：1051.3 µM ATP的濃度：10000 µM。 3. 實驗步驟 (1)採用Echo進行化合物稀釋，最終濃度為10 µM ~ 0.17 nM，取200 μL溶液加入到檢測板孔中； (2)將5 µL 4倍稀釋的GST-PRAK混合物加至檢測板孔中(含化合物)； (3)將15 µL 1.33倍稀釋的GST-P-p38α、ATP和Hsp27多肽混合物加至檢測板孔中(含化合物)； (4)1000轉/分，離心60秒左右，23℃溫育120分鐘； (5)加入60 µL IMAP溶液開始反應； (6)以1000轉/分離心檢測板約60秒，23℃溫育30分鐘； (7)在Neo2讀取檢測板(Ex/Em=485nm/FITC FP-P pol 528nm & FITC FP-S pol 528nm)； (8)通過訊號比值計算相對DMSO空白的相對酶活性抑制，利用軟體擬合曲線計算IC ₅₀值。 4. 實驗結果表4 代表性化合物對重組人p38α-PRAK激酶抑制活性及選擇性化合物編號 p38α-PRAK激酶抑制活性 IC ₅₀₍nM) 選擇性比值 (IC ₅₀PRAK/IC ₅₀MK2) Cpd-65A* 14.2 14.5 Cpd-78B* 2.9 0.17 Cpd-68A* 567 315 Cpd-81A* 2542 1338 Cpd-82A* 2316.0 503.5 Cpd-89D* 538 336 Cpd-91A* 4064.0 2257.8 Cpd-92D* 5677 366.3 Cpd-93A* 7267 865 註：*代表立體異構物 Test Example 3 Determination of the inhibitory activity of the representative compounds of the present invention on recombinant human p38 α -PRAK kinase and study of their selectivity 1. Experimental purpose: Determine the p38α-PRAK kinase inhibition IC50 value of the compound to evaluate the selectivity of the compound. 2. Experimental materials GST-PRAK solution concentration: 6710.15 nM GST-P-p38α solution concentration: 3030 nM Hsp27 peptide (FITC) concentration: 1051.3 µM ATP concentration: 10000 µM. 3. Experimental steps (1) Use Echo to dilute the compound to a final concentration of 10 µM ~ 0.17 nM, and add 200 µL of the solution to the wells of the test plate; (2) Add 5 µL of a 4-fold diluted GST-PRAK mixture to the wells of the test plate (containing the compound); (3) Add 15 µL of a 1.33-fold diluted GST-P-p38α, ATP and Hsp27 peptide mixture to the wells of the test plate (containing the compound); (4) Centrifuge at 1000 rpm for about 60 seconds and incubate at 23°C for 120 minutes; (5) Add 60 µL of IMAP solution to start the reaction; (6) Centrifuge the test plate at 1000 rpm for about 60 seconds and incubate at 23°C for 30 minutes; (7) Read the test plate on Neo2 (Ex/Em=485nm/FITC FP-P pol 528nm & FITC FP-S pol 528nm); (8) The relative enzyme activity inhibition relative to the DMSO blank was calculated by signal ratio, and the IC ₅₀ value was calculated by software fitting curve. 4. Experimental results Table 4 Inhibitory activity and selectivity of representative compounds on recombinant human p38α-PRAK kinase Compound No. p38α-PRAK kinase inhibitory activity IC _{50 (} nM) Selectivity ratio (IC ₅₀ PRAK/IC ₅₀ MK2) Cpd-65A* 14.2 14.5 Cpd-78B* 2.9 0.17 Cpd-68A* 567 315 Cpd-81A* 2542 1338 Cpd-82A* 2316.0 503.5 Cpd-89D* 538 336 Cpd-91A* 4064.0 2257.8 Cpd-92D* 5677 366.3 Cpd-93A* 7267 865 Note: * represents stereoisomers

結果表明，本發明代表性化合物對p38α下游的PRAK的抑制活性較弱，而對MK2抑制活性高，即本發明代表性化合物對p38α-MK2具有良好的選擇性。The results show that the representative compounds of the present invention have weak inhibitory activity against PRAK downstream of p38α, but have high inhibitory activity against MK2, that is, the representative compounds of the present invention have good selectivity for p38α-MK2.

測試例 4 本發明代表性化合物的藥代動力學研究1. 實驗目的以SD Rat大鼠為受試動物，應用LC/MS/MS法測定SD Rat大鼠經灌胃給予本發明化合物後不同時刻血漿中的藥物濃度。研究本發明化合物在SD Rat大鼠體內的藥代動力學行為，評價其藥動力學特徵 2. 實驗方案 (1)實驗動物 SD Rat大鼠15隻，雄性，分成5組，購自上海吉輝實驗動物飼養有限公司，動物生產許可證SCXK(滬)2017-0012。 (2)藥物配製處方為5%DMSO+5%聚氧乙烯蓖麻油+90%生理鹽水。先稱量適量受試陽性化合物CDD-450和本發明代表性化合物(折算純度和鹽係數)，加入處方量的DMSO，渦旋得到澄清透明溶液，再加入處方量的聚氧乙烯蓖麻油，渦旋混勻後再加入處方量的生理鹽水。得到0.6 mg/mL或2 mg/mL的溶液。配製過程中，如不能得到溶液，可嘗試不高於60℃水浴超音波幫助溶解。 (3)給藥大鼠禁食一夜後分別灌胃(2mpk)給藥。 (4)樣品採集給藥後15 min、30 min、1 h、2 h、4 h、6 h、8 h、24 h。在每個時間點採集約30 μL血液樣品置於含EDTA-K2抗凝劑的抗凝管中，30分鐘內離心得血漿。離心前全血樣品置於濕冰上。所有採集的血漿樣品保存在乾冰上或不高於-70℃中直至分析檢測。採用液相管柱層析-串聯質譜法(LC/MS/MS)測定血漿和給藥溶液中的原形藥物濃度。 3. 實驗結果本發明代表性化合物Cpd-68A/Cpd-81A/Cpd-89D/Cpd-93A及陽性化合物CDD450的SD Rat大鼠藥代動力學參數結果如表5及圖1-5所示。表5 本發明化合物及CDD-450的大鼠藥代動力學參數化合物編號 Cpd-68A Cpd-81A Cpd-89D Cpd-93A CDD-450 T _max(hr) 1.00 1.33 0.83 0.42 1.33 C _max(ng/mL) 741 1033 783 1140 415 T _1/2(hr) 2.8 2.29 2.61 2.74 1.78 AUC _last(hr*ng/mL) 4610 8047 4601 4340 1826 AUC _INF_pred(hr*ng/mL) 4621 8057 4818 4346 1919 MRT _INF_pred(h) 4.37 5.91 4.45 3.46 3.44 F(%) 119% 125% 109% 121% 71.3% Test Example 4 Pharmacokinetic Study of Representative Compounds of the Invention 1. Experimental Purpose SD rats were used as test animals. The LC/MS/MS method was used to determine the drug concentration in the plasma of SD rats at different times after oral administration of the compounds of the invention. The pharmacokinetic behavior of the compounds of the invention in SD rats was studied and their pharmacokinetic characteristics were evaluated. 2. Experimental Plan (1) Experimental Animals 15 male SD rats were divided into 5 groups and purchased from Shanghai Jihui Experimental Animal Breeding Co., Ltd., Animal Production License SCXK (Shanghai) 2017-0012. (2) The drug preparation formula was 5% DMSO + 5% polyoxyethylene castor oil + 90% saline. First, weigh an appropriate amount of the positive compound CDD-450 and the representative compound of the present invention (converted purity and salt coefficient), add the prescribed amount of DMSO, vortex to obtain a clear and transparent solution, then add the prescribed amount of polyoxyethylene castor oil, vortex and mix well, and then add the prescribed amount of physiological saline. A 0.6 mg/mL or 2 mg/mL solution is obtained. During the preparation process, if a solution cannot be obtained, try to use a water bath ultrasound at a temperature not higher than 60°C to assist in dissolution. (3) After fasting overnight, the rats were gavaged (2 mpk) and given the drug. (4) Sample collection was 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after drug administration. At each time point, about 30 μL of blood samples were collected and placed in anticoagulant tubes containing EDTA-K2 anticoagulant, and plasma was obtained by centrifugation within 30 minutes. Whole blood samples were placed on wet ice before centrifugation. All collected plasma samples were stored on dry ice or at a temperature not higher than -70°C until analysis and testing. Liquid column chromatography-tandem mass spectrometry (LC/MS/MS) was used to determine the concentration of the parent drug in plasma and the administration solution. 3. Experimental results The pharmacokinetic parameters of the representative compounds Cpd-68A/Cpd-81A/Cpd-89D/Cpd-93A and the positive compound CDD450 of the present invention in SD Rat rats are shown in Table 5 and Figures 1-5. Table 5 Pharmacokinetic parameters of the compounds of the present invention and CDD-450 in rats Compound No. Cpd-68A Cpd-81A Cpd-89D Cpd-93A CDD-450 _Tmax (hr) 1.00 1.33 0.83 0.42 1.33 Cmax ₍ ng/mL) 741 1033 783 1140 415 T _1/2 (hr) 2.8 2.29 2.61 2.74 1.78 AUC _last (hr*ng/mL) 4610 8047 4601 4340 1826 AUC _INF _pred (hr*ng/mL) 4621 8057 4818 4346 1919 MRT _INF _pred(h) 4.37 5.91 4.45 3.46 3.44 F(%) 119% 125% 109% 121% 71.3%

結論：本發明代表性化合物的藥代吸收良好，體內暴露量(AUC)及生物利用度(F%)等藥代性質全面優於陽性化合物CDD-450，表明本發明化合物具有良好的藥代吸收效果。Conclusion: The representative compounds of the present invention have good pharmacokinetic absorption, and their pharmacokinetic properties such as in vivo exposure (AUC) and bioavailability (F%) are better than those of the positive compound CDD-450, indicating that the compounds of the present invention have good pharmacokinetic absorption effect.

測試例 51. 實驗目的本次實驗目的是評價陽參化合物CDD450和本發明較佳化合物在佐劑誘導的大鼠關節炎症模型中的藥效作用，從而為之後的臨床研究提供臨床前藥效學相關資訊。 2. 縮寫表縮寫英語全名中文 AIA Adjuvant-induced arthritis 佐劑誘導的關節炎 H&E Hematoxylin-eosin staining 蘇木精-伊紅染色法 PO Peros 口服給藥 bid or BID Bis in die 每天兩次 IACUC Association for Assessment and Accreditation of Laboratory Animal Care 實驗動物福利與使用管理委員會 mg/kg Milligrams per kiligram of body weight 毫克每千克體重 qd or QD quaque die 每天一次 SPF Specific Pathogen Free 無特定病原體 n or N Number 數量 ℃ Centigrade 攝氏度 AUC Area under the curve 曲線下面積 SEM Standard Error of the Mean 標準誤 3. 實驗材料 3.1 實驗試劑結核分枝桿菌H37Ra，Difico(Detroit,MI,USA)，貨號：231141；石蠟油，國藥集團，貨號：30139828；甲基纖維素，上海阿拉丁生化科技股份有限公司，CAS號：9004-67-5； Tween-20，Sigma，貨號：90005-64-5。 3.2 實驗儀器分析天平：Sartorious，CPA225D；稱量天平：常州天之平電子天平，YH-2000；體積測量儀：Italy UGO BASILE，Biological Research Apparatus 21025。 4. 實驗方法 4.1 實驗動物及飼養環境動物品系： Lewis大鼠供應商：北京維通利華實驗動物有限公司性別和體重：雌性，170-190g 飼養地：南通藥明康得SPF動物房適應期： ≥7天環境： SPF動物飼養室溫度： 20-26 ℃ 濕度： 40-70% 光照：明(08:00-20:00)、暗(20:00-08:00)各12小時飼養密度： 3隻/籠食物：自由進食(輻射滅菌飼料) 水：自由飲水(超純水器製備) 4.2 實驗方法 4.2.1 造模及給藥 (1)佐劑配製：稱取結核分枝桿菌H37Ra 100 mg，研磨約5分鐘，石蠟油清洗研缽3次，終濃度為10 mg/mL。超音波破碎，冰水混合物中超音波約30分鐘。 (2)關節炎的誘導：將佐劑震盪混勻，用1毫升的玻璃注射器(20G針頭)抽取，再換成25G針頭注射。免疫前不停地轉動注射器，以免結核分枝桿菌沉澱。用異氟烷麻醉大鼠後，在大鼠左腳腳掌皮下注射0.1 mL佐劑，正常組注射0.1 mL石蠟油。第一次注射佐劑當天為第0天。 (3)給藥和劑量設計：在第13天，所有動物均顯現足部紅斑或紅腫等關節炎症狀，根據實驗方案，按照關節炎評分和體重分組。分組及每組的給藥劑量如表6所示，灌胃給藥體積為5 mL/kg，受試藥物組和Vehicle組大鼠每天給藥兩次，共持續14天。表6. 藥效學實驗分組及劑量設計組別數量(n) 受試物劑量(mg/kg) 給藥途徑給藥計畫 G1 4 Normal - - - G2 6 Vehicle - p.o. bid × 14d G3 7 CDD450 1 p.o. bid × 14d G4 7 CDD450 3 p.o. bid × 14d G5 7 CDD450 9 p.o. bid × 14d G6 7 較佳化合物 0.3 p.o. bid × 14d G7 7 較佳化合物 1 p.o. bid × 14d G8 7 較佳化合物 3 p.o. bid × 14d (4)關節炎發病指標測定：a：給藥後每週稱體重三次。b：給藥後每週測量足體積三次。c：給藥後每週評分三次。根據病變的不同程度(紅腫，關節變形)按照0~4分的標準進行評分，每個肢體的最高評分為4分，每隻動物最高評分為12分(模型誘導的左後肢除外)。評分標準如表7所示。表7. 關節炎臨床評分標準分值臨床症狀 0 無紅斑和紅腫 1 近跗骨附近或踝關節或蹠骨出現紅斑或輕度紅腫或有一個腳趾有紅斑和紅腫 2 踝關節和蹠骨輕微紅斑和腫脹，有兩個或兩個以上腳趾有紅腫和紅斑 3 踝、腕關節和蹠骨中度紅斑和腫脹 4 踝、腕關節，蹠骨和腳趾全部嚴重紅腫 4.2.2 樣品收集及病理檢測實驗終點，安樂死大鼠。取大鼠右後肢，用多聚甲醛溶液浸泡，用甲酸溶液脫鈣，石蠟包埋，切片，H&E染色，番紅染色，顯微鏡觀察。從炎症細胞浸潤、血管翳生成、軟骨損傷和骨吸收等四個方面對關節的損傷程度進行評價，並按照0~4分的標準進行評分，並提供每組一張代表性照片。各項評分標準如表8所示。表8. 關節炎病理學評分標準病變病變特點分值炎性細胞浸潤無可見炎性細胞 0 滑膜下細胞纖維化，並有極少量細胞浸潤 1 滑膜細胞增生，少量單核細胞浸潤 2 滑膜細胞增生，大量單核細胞、漿細胞、淋巴細胞浸潤 3 關節周圍大量炎性細胞浸潤，組織纖維化，滑膜增厚 4 血管翳形成無可見血管翳形成 0 軟骨邊緣有極少量血管翳形成 1 軟骨間纖維組織增生，關節邊緣有少量血管翳形成 2 50%的關節軟骨表面有血管翳形成 3 整個關節軟骨表面可見血管翳形成 4 軟骨損傷無可見軟骨損傷 0 關節軟骨細胞增生 1 軟骨細胞基質丟失，少量軟骨細胞被破壞 2 關節周圍纖維組織增生，大量軟骨細胞被破壞 3 關節軟骨間有大量纖維組織增生，軟骨糜爛 4 骨再吸收無可見骨再吸收 0 滑膜邊緣可見極少量的骨再吸收 1 小範圍骨組織可見少量破骨細胞形成， 2 局部關節軟骨下的骨組織伴有骨再吸收 3 大範圍的骨組織發生骨再吸收，並伴有軟骨糜爛 4 4.3 統計學處理實驗資料利用Graph Pad Prism 9軟體進行統計分析及作圖，用平均數±標準誤表示(Mean ± SEM)，兩組間比較採用 t檢驗，多組間分析選用Dunnett雙因素方差分析(Two -way ANOVA)， P＜ 0.05表示具有統計學意義。 5. 實驗結果 5.1關節炎臨床評分本實驗評價了化合物在大鼠關節炎(AIA)模型中對臨床評分的改善作用。通過在實驗第0天在大鼠左腳腳掌皮下注射0.1 mL佐劑誘導構建大鼠AIA模型，免疫後第13天，大鼠開始出現關節炎臨床症狀。第13天開始給藥，Vehicle組的平均臨床評分逐漸升高，至第27天達到9.00分，提示佐劑誘導的關節炎模型的成功建立。臨床評分最後一天(第27天)時，全部受試物都對大鼠關節炎臨床評分有抑制作用，其中較佳化合物的3 mg/kg BID組對關節炎大鼠臨床評分降低至4以下。與Vehicle組的臨床評分相比，所有受試藥物組在Day27臨床評分降低( P＜0.0001)。通過分析每組每隻動物的臨床評分曲線，計算曲線下面積AUC，通過組間AUC平均值(參見圖6)，計算各給藥組相對於Vehicle組的抑制率，其中受試物CDD450的9 mg/kg BID組，較佳化合物的1 mg/kg BID組和3 mg/kg BID組的抑制率分別為47.1%，46.7%和55.2%。計算公式如下： Test Example 5 1. Experimental Purpose The purpose of this experiment is to evaluate the pharmacodynamic effects of the Yangshen compound CDD450 and the preferred compounds of the present invention in the adjuvant-induced rat joint inflammation model, so as to provide preclinical pharmacodynamics-related information for subsequent clinical studies. 2. Abbreviation Table Abbreviation Full English name Chinese AIA Adjuvant-induced arthritis Adjuvant-induced arthritis H&E Hematoxylin-eosin staining Hematoxylin-eosin staining PO Peros Oral medication bid or BID Bis in die Twice a day IACUC Association for Assessment and Accreditation of Laboratory Animal Care Committee on the Welfare and Use of Laboratory Animals mg/kg Milligrams per kiligram of body weight mg/kg body weight qd or QD quaque die Once a day SPF Specific Pathogen Free Specific Pathogen Free n or N Number quantity ℃ Centigrade Celsius AUC Area under the curve Area under the curve SEM Standard Error of the Mean Standard error 3. Experimental Materials 3.1 Experimental Reagents Mycobacterium tuberculosis H37Ra, Difico (Detroit, MI, USA), Catalog No.: 231141; Paraffin oil, Sinopharm Group, Catalog No.: 30139828; Methylcellulose, Shanghai Aladdin Biochemical Technology Co., Ltd., CAS No.: 9004-67-5; Tween-20, Sigma, Catalog No.: 90005-64-5. 3.2 Experimental Instruments Analytical balance: Sartorious, CPA225D; Weighing balance: Changzhou Tianzhiping electronic balance, YH-2000; Volumetric instrument: Italy UGO BASILE, Biological Research Apparatus 21025. 4. Experimental Methods 4.1 Experimental Animals and Housing Environment Animal series: Lewis rats Supplier: Beijing Weitonglihua Experimental Animal Co., Ltd. Gender and weight: Female, 170-190g Farming land: Nantong WuXi AppTec SPF Animal House Adaptation period: ≥ 7 days environment: SPF Animal Housing temperature: 20-26 ℃ Humidity: 40-70% illumination: Daylight (08:00-20:00), Darkness (20:00-08:00) 12 hours each Stocking density: 3/cage food: Free access to food (irradiated sterilized feed) water: Free drinking water (prepared by ultra-pure water maker) 4.2 Experimental methods 4.2.1 Modeling and drug administration (1) Preparation of adjuvant: Weigh 100 mg of Mycobacterium tuberculosis H37Ra, grind for about 5 minutes, clean the mortar 3 times with paraffin oil, and the final concentration is 10 mg/mL. Ultrasonic fragmentation, ultrasonic in ice-water mixture for about 30 minutes. (2) Induction of arthritis: Shake the adjuvant to mix, draw out with a 1 ml glass syringe (20G needle), and then replace it with a 25G needle for injection. Turn the syringe continuously before immunization to prevent Mycobacterium tuberculosis from precipitating. After anesthetizing the rats with isoflurane, 0.1 mL of adjuvant was injected subcutaneously into the sole of the left foot of the rats, and 0.1 mL of paraffin oil was injected into the normal group. The day of the first injection of adjuvant was day 0. (3) Drug administration and dosage design: On the 13th day, all animals showed arthritis symptoms such as erythema or swelling on the feet. According to the experimental plan, they were divided into groups according to the arthritis score and body weight. The groups and the dosage of each group are shown in Table 6. The oral administration volume was 5 mL/kg. The rats in the test drug group and the vehicle group were given the drug twice a day for a total of 14 days. Table 6. Grouping and dosage design of pharmacodynamic experiment Group Quantity(n) Test substance Dosage (mg/kg) Route of administration Medication plan G1 4 Normal - - - G2 6 Vehicle - po bid × 14d G3 7 CDD450 1 po bid × 14d G4 7 CDD450 3 po bid × 14d G5 7 CDD450 9 po bid × 14d G6 7 Preferred compounds 0.3 po bid × 14d G7 7 Preferred compounds 1 po bid × 14d G8 7 Preferred compounds 3 po bid × 14d (4) Determination of arthritis pathogenesis indicators: a: Weigh three times a week after medication. b: Measure foot volume three times a week after medication. c: Score three times a week after medication. Score according to the degree of lesions (redness, swelling, joint deformity) according to the standard of 0 to 4 points. The highest score for each limb is 4 points, and the highest score for each animal is 12 points (except for the left hind limb induced by the model). The scoring criteria are shown in Table 7. Table 7. Clinical scoring criteria for arthritis Points Clinical symptoms 0 No erythema or swelling 1 Red spots or mild swelling near the tarsal bones, ankle joints, or metatarsals, or red spots and swelling on one toe 2 Mild erythema and swelling of the ankle joint and metatarsal bones, with redness and swelling of two or more toes 3 Moderate erythema and swelling of ankles, wrists, and metatarsals 4 Severe swelling and redness of the ankles, wrists, metatarsals, and toes 4.2.2 Sample collection and pathological examination At the end of the experiment, the rats were euthanized. The right hind limbs of the rats were taken, soaked in paraformaldehyde solution, decalcified with formic acid solution, embedded in paraffin, sliced, stained with H&E, stained with safranin, and observed under a microscope. The degree of joint damage was evaluated from four aspects: inflammatory cell infiltration, pannus formation, cartilage damage, and bone resorption. The scores were scored according to the standard of 0 to 4 points, and a representative photo was provided for each group. The scoring standards for each item are shown in Table 8. Table 8. Pathological scoring standards for arthritis Pathological changes Pathological characteristics Points Inflammatory cell infiltration No inflammatory cells were seen 0 Subsynovial cells are fibrotic with minimal cellular infiltration 1 Synovial cell proliferation, small amount of mononuclear cell infiltration 2 Synovial cell hyperplasia, infiltration of a large number of monocytes, plasma cells, and lymphocytes 3 There is a large amount of inflammatory cell infiltration around the joint, tissue fibrosis, and synovial thickening. 4 Pannus formation No pannus formation was visible 0 There is a very small amount of pannus formation at the edge of the cartilage 1 There is proliferation of fibrous tissue between the cartilage and a small amount of pannus formation at the edge of the joint. 2 50% of the articular cartilage surface has pannus formation 3 Pannus formation can be seen on the entire surface of the articular cartilage 4 Cartilage damage No visible cartilage damage 0 Articular chondrocyte hyperplasia 1 The chondrocyte matrix is lost and a small number of chondrocytes are destroyed. 2 The fibrous tissue around the joint proliferates and a large number of cartilage cells are destroyed. 3 There is a large amount of fibrous tissue proliferation between the joint cartilages, and the cartilage is eroded. 4 Bone resorption No bone resorption 0 Minimal bone resorption is seen at the synovial margin 1 A small amount of osteoclasts can be seen in a small area of bone tissue. 2 Local subchondral bone tissue with bone resorption 3 Extensive bone resorption with cartilage erosion 4 4.3 Statistical analysis Experimental data were analyzed and plotted using Graph Pad Prism 9 software, with mean ± SEM. The t- test was used for comparison between two groups, and Dunnett's two-way ANOVA was used for analysis between multiple groups. P < 0.05 was considered statistically significant. 5. Experimental results 5.1 Clinical score of arthritis This experiment evaluated the effect of the compound on the improvement of clinical scores in the rat arthritis (AIA) model. The rat AIA model was established by subcutaneously injecting 0.1 mL of adjuvant into the sole of the left foot of the rat on day 0 of the experiment. On day 13 after immunization, the rats began to show clinical symptoms of arthritis. Drug administration began on the 13th day, and the average clinical score of the Vehicle group gradually increased, reaching 9.00 points on the 27th day, indicating that the adjuvant-induced arthritis model was successfully established. On the last day of clinical evaluation (day 27), all test substances had an inhibitory effect on the clinical score of rat arthritis, and the 3 mg/kg BID group of the best compound reduced the clinical score of arthritis rats to below 4. Compared with the clinical score of the Vehicle group, the clinical scores of all test drug groups on Day 27 were reduced ( P < 0.0001). By analyzing the clinical score curve of each animal in each group, the area under the curve AUC was calculated, and the inhibition rate of each drug administration group relative to the Vehicle group was calculated by the average AUC between groups (see Figure 6). The inhibition rates of the 9 mg/kg BID group of the test substance CDD450, the 1 mg/kg BID group of the better compound, and the 3 mg/kg BID group were 47.1%, 46.7%, and 55.2%, respectively. The calculation formula is as follows:

結論：本發明的較佳化合物在大鼠關節炎炎症模型中同劑量下的治療效果顯著優於陽參化合物CDD450，大鼠後足CT圖片(參見圖7)顯示較佳化合物體內治療效果非常突出，其在3 mg/kg劑量下的治療效果優於陽參化合物9 mg/kg劑量的治療效果。綜上，本發明的較佳化合物具有出色的抗炎作用。Conclusion: The therapeutic effect of the preferred compound of the present invention at the same dose in the rat arthritis inflammation model is significantly better than that of the Yangshen compound CDD450. The CT image of the rat hind foot (see Figure 7) shows that the in vivo therapeutic effect of the preferred compound is very prominent, and its therapeutic effect at a dose of 3 mg/kg is better than that of the Yangshen compound at a dose of 9 mg/kg. In summary, the preferred compound of the present invention has an excellent anti-inflammatory effect.

測試例 61. 實驗目的評價SD大鼠每天1次連續14天經口灌胃給予本發明較佳化合物後可能引起毒性反應的性質、程度、量效和時效關係，同時研究其毒代動力學特徵，瞭解毒性研究中暴露劑量與毒理學結果之間的關係，為後續試驗設計提供參考。 2. 實驗方法 66隻健康適用的SD大鼠，SPF級，雌雄各半，雄性動物體重範圍為248.2-270.1 g，雌性動物體重範圍為180.1-205.4 g。按性別和體重隨機分為13組，第1-7組為毒性實驗組，每組每性別3隻動物；第8-13組為毒代實驗組，每組每性別2隻動物。實驗期間所有動物每天1次，連續14天，經口灌胃給於溶劑或受試物，第1組為溶劑對照組；第2/8、3/9、4/10組給於受試物為陽參CDD450；第5/11、6/12、7/13組給於本發明較佳化合物。給藥體積均為10 mL/kg，給藥劑量如下：第1組：動物給藥劑量為0 mg/kg(溶劑對照)；第2/8、3/9、4/10組：雄性動物給藥劑量分別為10 mg/kg、30 mg/kg和60 mg/kg，雌性動物給藥劑量分別為6 mg/kg、12 mg/kg和30 mg/kg；第5/11、6/12、7/13組：雄性動物給藥劑量分別為6 mg/kg、20 mg/kg和60 mg/kg，雌性動物給藥劑量分別為3 mg/kg、10 mg/kg和30 mg/kg。適應期，所有動物於Day 1進行1次詳細的臨床觀察、體重稱量、每天進行2次籠邊觀察。實驗期間，所有存活動物每天進行2次籠邊觀察(計畫解剖的動物於解剖當天進行1次籠邊觀察)；第1-7組所有存活動物在Day 1-Day 14每天給藥後觀察一次，第8-13組動物僅在籠邊觀察出現異常時進行詳細臨床觀察；實驗期間對第1-13組所有存活動物於Day 1、Day 4、Day 7、Day 10和Day 14給藥前各稱重1次，計畫解剖動物在解剖前(Day 15)稱重1次；測定第1-7組動物在Day 2-Day 3、Day 6 - Day 7、Day 10 - Day 11和Day 13-Day 14的耗食量並記錄；且計畫解剖(Day15)的存活動物進行臨床病理學檢查(包括：血液學、凝血、血清生化和尿液分析)。臨檢樣本採集前所有動物(第1-7組)禁食過夜(12.5 h)但不禁水，於Day 15肌肉注射舒泰麻醉後經腹主動脈穿刺採集血樣(大約4.8 mL)，用於血液學、凝血和血清生化學分析。解剖及大體觀察，第1-7組每組存活動物在給藥期末進行剖檢，同時取組織器官稱重並計算臟器係數。第8-13組動物於首次給藥Day 1、Day 7和末次給藥Day 14各採集1輪毒代動力學血樣，Day 1：共8個採集時間點，分別為給藥後 0.25 h、0.5 h、1 h、2 h、4 h、6 h、8 h 和 24 h(下次給藥前)；Day7和Day 14共9個採集時間點，分別為給藥前，給藥後0.25 h、0.5 h、1 h、2 h、4 h、6 h、8 h和24 h。用已驗證過的LC-MS/MS分析方法檢測血漿中陽參CDD450和較佳化合物的濃度，採用Phoenix WinNonlin ^®7.0非房室模型計算藥物的AUC _(0-t)、C _max和T _max。第8-13組動物，在末次給藥採樣結束後被安樂死。 1. 實驗結論在本試驗條件下，未觀察到與供試品相關的毒性反應。 Test Example 6 1. Experimental Purpose To evaluate the nature, degree, dose-effect and time-effect relationship of the toxicity reaction that may be caused by oral gavage of the preferred compound of the present invention to SD rats once a day for 14 consecutive days, and to study its toxicokinetic characteristics at the same time, to understand the relationship between the exposure dose and toxicological results in toxicity studies, and to provide a reference for the subsequent test design. 2. Experimental Methods 66 healthy and suitable SD rats, SPF grade, half male and half female, the body weight range of male animals is 248.2-270.1 g, and the body weight range of female animals is 180.1-205.4 g. They were randomly divided into 13 groups according to sex and weight. Groups 1-7 were toxicity test groups, with 3 animals of each sex in each group; Groups 8-13 were toxicokinetic test groups, with 2 animals of each sex in each group. During the experiment, all animals were given the solvent or the test substance by oral gavage once a day for 14 consecutive days. Group 1 was the solvent control group; Groups 2/8, 3/9, and 4/10 were given the test substance Yangshen CDD450; and Groups 5/11, 6/12, and 7/13 were given the preferred compounds of the present invention. The volume of drug administration was 10 mL/kg, and the dosage was as follows: Group 1: The animal dosage was 0 mg/kg (solvent control); Groups 2/8, 3/9, and 4/10: The dosages of male animals were 10 mg/kg, 30 mg/kg, and 60 mg/kg, respectively, and the dosages of female animals were 6 mg/kg, 12 mg/kg, and 30 mg/kg, respectively; Groups 5/11, 6/12, and 7/13: The dosages of male animals were 6 mg/kg, 20 mg/kg, and 60 mg/kg, respectively, and the dosages of female animals were 3 mg/kg, 10 mg/kg, and 30 mg/kg, respectively. During the adaptation period, all animals underwent a detailed clinical observation and body weight measurement on Day 1, and cage-side observations were conducted twice a day. During the experiment, all surviving animals were observed twice a day at the cage side (animals scheduled for dissection were observed once on the day of dissection); all surviving animals in groups 1-7 were observed once every day after drug administration from Day 1 to Day 14, and animals in groups 8-13 were only subject to detailed clinical observation when abnormalities were observed at the cage side; during the experiment, all surviving animals in groups 1-13 were weighed once before drug administration on Day 1, Day 4, Day 7, Day 10, and Day 14, and animals scheduled for dissection were weighed once before dissection (Day 15); the blood pressure of animals in groups 1-7 was measured on Day 2-Day 3, Day 6 - Day 7, Day 10 - Day 11, and Day 13-Day The food consumption on Day 14 was recorded; and the surviving animals planned for autopsy (Day 15) were subjected to clinical pathological examination (including hematology, coagulation, serum biochemistry and urine analysis). Before the collection of clinical samples, all animals (Groups 1-7) were fasted overnight (12.5 h) but not water. Blood samples (approximately 4.8 mL) were collected by abdominal aortic puncture after intramuscular injection of Zota anesthesia on Day 15 for hematology, coagulation and serum biochemistry analysis. Autopsy and gross observation: The surviving animals in each group of Groups 1-7 were autopsied at the end of the drug administration period, and tissues and organs were weighed and the organ coefficients were calculated. Animals in groups 8-13 were collected for kinetic blood sampling on Day 1, Day 7, and Day 14, the first dosing and the last dosing. On Day 1, there were 8 sampling time points, including 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after dosing (before the next dosing); on Day 7 and Day 14, there were 9 sampling time points, including before dosing, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after dosing. The concentrations of CDD450 and the best compounds in plasma were detected by a validated LC-MS/MS analytical method, and the AUC _(0-t) , C _max , and T _max of the drugs were calculated using the Phoenix WinNonlin ^® 7.0 non-compartmental model. Animals in groups 8-13 were euthanized after the last dosing and sampling. 1. Experimental conclusions Under the conditions of this study, no toxic reactions related to the test article were observed.

以上對本發明技術方案的實施方式進行了示例性的說明。應當理解，本發明的保護範圍不拘囿於上述實施方式。凡在本發明的精神和原則之內，本發明所屬技術領域中具有通常知識者所做的任何修改、均等替換、改進等，均應包含在本發明專利範圍的保護範圍之內。The above is an exemplary description of the implementation of the technical solution of the present invention. It should be understood that the protection scope of the present invention is not limited to the above implementation. Any modification, equal replacement, improvement, etc. made by a person with ordinary knowledge in the technical field to which the present invention belongs within the spirit and principles of the present invention shall be included in the protection scope of the patent scope of the present invention.

無without

圖1. 化合物Cpd-68A的藥代曲線圖；圖2. 化合物Cpd-81A的藥代曲線圖；圖3. 化合物Cpd-93A的藥代曲線圖；圖4. 化合物Cpd-89D的藥代曲線圖；圖5. 化合物CDD-450的藥代曲線圖；圖6. 較佳化合物和陽參化合物CDD450臨床評分曲線下面積(AUC)柱狀圖；圖7. 大鼠經較佳化合物與陽參化合物CDD450治療後的後足micro CT圖。 Figure 1. Pharmacokinetic curve of compound Cpd-68A; Figure 2. Pharmacokinetic curve of compound Cpd-81A; Figure 3. Pharmacokinetic curve of compound Cpd-93A; Figure 4. Pharmacokinetic curve of compound Cpd-89D; Figure 5. Pharmacokinetic curve of compound CDD-450; Figure 6. Area under the clinical score curve (AUC) bar graph of the best compound and Yangshen compound CDD450; Figure 7. Micro CT images of the hind feet of rats treated with the best compound and Yangshen compound CDD450.

Claims

A compound represented by formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof: Formula I wherein, X is selected from CH or N; Y is selected from a chemical bond, a C _1-6 alkyl group, a deuterated C _1-6 alkyl group, a 4-7 membered heterocyclic group; Ring A and Ring B are the same or different and are independently selected from C _6-14 aryl group, a 5-14 membered heteroaryl group, a 3-14 membered heterocyclic group, a C _3-12 cycloalkyl group; each R is the same or different and is independently selected from halogen, CN, hydroxyl, amine, C _1-6 alkyl group, C _1-6 alkoxy group, halogenated C _1-6 alkyl group, halogenated C _1-6 alkoxy group, C _3-8 cycloalkyl group, -C(R ₁ )(R ₂ )(R ₃ ), -P(═O)(R ₂₁ )(R ₂₂ ); wherein, R ₁ , R _{R 2} is the same or different and is independently selected from H, deuterium, halogen, CN, hydroxyl, amine, C _1-6 alkyl, C _1-6 alkoxy, deuterated C _1-6 alkyl, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _3-8 cycloalkyl; or, R ₁ , R ₂ together with the atoms to which they are attached form the following group optionally substituted by 1, 2 or more R': C _3-8 cycloalkyl or 3-10 membered heterocyclic group; each R' is the same or different and is independently selected from H, deuterium, halogen, CN, pendoxy (=O), C _1-6 alkyl, C _1-6 alkoxy; R ₃ is selected from H, deuterium, halogen, CN, hydroxyl, amine, C 1-6 alkyl, C 1-6 alkoxy, deuterated C _1-6 alkyl, halogenated C _1-6 alkoxy, C 3-8 cycloalkyl _wherein the at least one R 4 and R ₇ are _the same or different _and are independently selected from H, deuterium, halogen, _CN _, _hydroxyl _, _amino _, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, _C _3-8 _cycloalkyl ; R ₅ and R ₆ are the same or different _and are independently selected from H, deuterium, halogen, CN, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl _{, halogenated C 1-6} _alkoxy _, C _3-8 cycloalkyl; R ₈ , R ₉ , R ₁₀ , R ₁₁ , and R ₁₂ are the same _or different and are independently selected from H, deuterium, or the following groups which are unsubstituted or optionally substituted with 1 _, ₂ or more Ra: C _1-6 alkyl, C _1-6 alkoxy, or C _3-8 cycloalkyl; each Ra is the same or different and is independently selected from deuterium, halogen, CN, amine, hydroxyl, pendoxy (=O), C _1-6 alkyl, _{or C 1-6} _alkoxy ; or R ₉ , R _{R 10} together with the N atom to which they are attached form the following group which is unsubstituted or optionally substituted by 1, 2 or more R b : a 3-10 membered heterocyclic group or a 5-10 membered heteroaryl group; each R b is the same or different and is independently selected from deuterium, a halogen, CN, a pendoxy group (=O), a hydroxyl group, an amine group, a C _1-6 alkyl group, a C _1-6 alkoxy group; R ₂₁ and R ₂₂ are the same or different and are independently selected from H, deuterium, a C _1-6 alkyl group, a C _1-6 alkoxy group, a halogenated C _1-6 alkyl group, a halogenated C _1-6 alkoxy group, a C _3-8 cycloalkyl group; r is selected from 1, 2 or 3; p and q are the same or different and are independently selected from 0, 1, 2, 3 or 4; m is selected from an integer of 0-6; n is selected from 0, 1 or 2.

The compound as claimed in claim 1, wherein X is selected from N; preferably, X is selected from CH; when X is CH, _R7 may replace H at that position, preferably, _XR7 is selected from C-CN; preferably, Y is selected from _C1-3 alkyl, deuterated _C1-3 alkyl, _4-6 membered heterocyclic group; preferably, Y is selected from methylene, deuterated methylene ( _-CD2- or -CHD-), ethylene (-CH( _CH3 )- or _-CH2CH2- ), -C( _CH3 ) _2- , piperidinyl, piperazinyl, tetrahydropyrrolyl, azacyclobutyl; preferably, Y is selected from _-CH2- , _-CD2- , -CH( _CH3 )-, -C( _CH3 ) _2- or azacyclobutyl; Preferably, ring A and ring B are the same or different and are independently selected from C _6-10 aryl groups and 5-10 membered heteroaryl groups; Preferably, ring A and ring B are the same or different and are independently selected from phenyl groups or 5-6 membered heteroaryl groups containing 1-3 heteroatoms selected from N, O and S; Preferably, ring A is selected from pyridyl or phenyl; Preferably, ring B is selected from pyrimidinyl, pyrazinyl, pyridyl, pyridine nitrogen oxide or phenyl.

The compound of claim 1 or 2, wherein each R is the same or different and is independently selected from halogen, CN, -C(R ₁ )(R ₂ )(R ₃ ) or -P(=O)(R ₂₁ )(R ₂₂ ); preferably, R ₁ and R ₂ are the same or different and are independently selected from C _1-6 alkyl and C _3-8 cycloalkyl; or, R ₁ and R ₂ together with the atoms to which they are attached form a C _3-8 cycloalkyl; preferably, R ₁ and R ₂ are independently selected from methyl and cyclopropyl; or, R ₁ and R ₂ together with the atoms to which they are attached form a cyclopropyl, cyclobutyl or cyclopentyl; preferably, R ₃ is selected from CN, hydroxyl, amine, -C(O)R ₈ , -C(O)NR ₉ R ₁₀ , -S(O) ₂ -R ₁₁ , -COOR ₁₂ ; preferably, R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ are the same or different and are independently selected from H, C _1-6 alkyl, C _3-8 cycloalkyl; or R ₉ , R ₁₀ together with the N atom to which they are connected form the following group which is unsubstituted or optionally substituted by 1, 2 or more R b : a 3-8 membered heterocyclic group; each R b is the same or different and is independently selected from halogen, CN, pendoxy (=O), hydroxyl, C _1-6 alkyl, C 1-6 alkoxy; preferably, R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ are the same or different and are independently selected from H, C _1-3 alkyl, C _3-8 cycloalkyl; R ₉ , R ₁₀ together with the N atom to which they are attached form the following groups which are unsubstituted or optionally substituted with 1, 2 or more R b : _3-6 membered heterocyclic group; each R b is the same or different and is independently selected from hydroxyl, C _1-6 alkyl; preferably, R ₈ , R ₁₁ are independently selected from methyl; preferably, R ₉ , R ₁₀ are the same or different and are independently selected from H, methyl, ethyl, isopropyl, cyclopropyl; or R ₉ , R ₁₀ together with the N atom to which they are attached form an azacyclobutyl, piperazinyl substituted with 1 or 2 R b ; each R b is the same or different and is independently selected from hydroxyl or methyl; preferably, R ₉ , R _{R 10} are the same or different and are independently selected from H, methyl, ethyl, isopropyl, cyclopropyl; or R ₉ , R ₁₀ together with the N atom to which they are connected form 3-hydroxy-1-azacyclobutyl or 4-methylpiperazin-1-yl; preferably, R ₃ is selected from CN, hydroxyl, amine, -CONH ₂ , -SO ₂ CH ₃ , -COCH ₃ , -CONHCH ₃ , -CONHCH ₂ CH ₃ , -CONHCH(CH ₃ ) ₂ , -CONH-cyclopropyl, -CON(CH ₃ ) ₂ , , , -C(CH ₃ ) ₂ (OH), -COOH, -COOCH ₃ , -COOC ₂ H ₅ ; preferably, R ₂₁ and R ₂₂ are the same or different and are independently selected from H, deuterium, C _1-6 alkyl, C _1-6 alkoxy; preferably, R ₂₁ and R ₂₂ are both selected from methyl.

The compound as described in any one of claims 1 to 3, wherein each R ₄ and R ₇ are the same or different and are independently selected from halogen, CN, and C _1-6 alkyl; preferably, each R ₄ and R ₇ are the same or different and are independently selected from halogen, CN, and methyl; preferably, R ₄ is selected from methyl or F; preferably, R ₇ is selected from F or CN; preferably, R ₅ and R ₆ are the same or different and are independently selected from halogen, CN, and C _1-6 alkyl; preferably, R ₅ and R ₆ are the same or different and are independently selected from halogen and methyl; preferably, R ₅ is selected from methyl; preferably, R ₆ is selected from Cl, Br or methyl; preferably, r is selected from 1 or 2; preferably, p is selected from 2 or 3; Preferably, q is selected from 1 or 2; Preferably, m is selected from 0 or 1; Preferably, n is selected from 2.

The compound as described in any one of claims 1 to 4, wherein the compound represented by formula I is selected from the structures shown below: , , Formula IA Formula IA-1 Formula IA-2 wherein X, Y, Ring A, Ring B, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, q have the definitions as described in any one of claims 1 to 4; preferably, the compound represented by Formula I is selected from the structures shown below: Formula IB wherein X, Y, R, R ₄ , R ₅ , R ₆ , R ₇ , r, p, q have the definitions as described in any one of claims 1 to 4, Z ₁ , Z ₂ , Z ₃ are the same or different and are independently selected from N, N(O) or CH; Preferably, Z ₁ is selected from N, Z ₂ is selected from N, N(O) or CH, and Z ₃ is selected from N, N(O) or CH; Preferably, Z ₁ is selected from CH, Z ₂ is selected from N, N(O) or CH, and Z ₃ is selected from N, N(O) or CH; Preferably, the compound represented by Formula I is selected from the structures shown below: Formula IB-1 wherein X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, q have the definitions as described in any one of claims 1 to 4; Z ₁ , Z ₂ , Z ₃ are the same or different and are independently selected from N, N(O) or CH; Preferably, the compound represented by Formula I is selected from the structures shown below: Formula I-1 Formula I-2 Formula I-3 Formula I-4 Formula I-5 Formula I-6 Formula I-7 Formula I-8 Formula I-9 Formula I-10 wherein X, Y, Ring A, Ring B, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , p, q have the definitions as described in any one of claims 1 to 4; preferably, the compound represented by Formula I is selected from the structures shown below: Formula IC wherein X, Y, R, R ₄ , R ₅ , R ₆ , R ₇ , r, p, q have the definitions as described in any one of claims 1 to 4; preferably, the compound represented by formula I is selected from the structures shown below: Formula II Formula II-1 Formula II-2 Wherein, Y, R ₁ , R ₂ , R ₃ , R ₄ , q have the definitions as described in any one of claims 1 to 4; Preferably, the compound represented by Formula I is selected from the structures shown below: Formula III Formula III-1 wherein R ₁ , R ₂ , R ₄ , R ₉ , R ₁₀ , Y, and q have the definitions as described in any one of claims 1 to 4; preferably, the compound represented by formula I is selected from the structures shown below: Formula IV wherein R ₉ and R ₁₀ have the definitions as described in any one of claims 1 to 4; Preferably, the compound represented by formula I is selected from the structures shown below: Type V Type V-1 Type V-2 Formula V-3 wherein Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , R, p, q have the definitions as described in any one of claims 1 to 4; preferably, the compound represented by formula I is selected from the structures shown below: Formula VI Formula VI-1 Formula VI-2 Formula VI-3 wherein Y, R ₄ , R ₆ , R, q have the definitions as described in any one of claims 1 to 4; wherein Z ₁ , Z ₂ , Z ₃ are the same or different and are independently selected from N, N(O) or CH; preferably, Z ₁ is selected from N, Z ₂ is selected from N, N(O) or CH, and Z ₃ is selected from N, N(O) or CH; preferably, Z ₁ is selected from CH, Z ₂ is selected from N, N(O) or CH, and Z ₃ is selected from N, N(O) or CH; preferably, the compound represented by formula I is selected from the structures shown below: Formula VII wherein Y, R ₄ , R ₆ , and R have the definitions as described in any one of claims 1 to 4.

The compound as described in any one of claims 1 to 5, wherein the compound represented by formula I is selected from the structures shown below: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , ; Preferably, the compound represented by formula I is selected from the structures shown below: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

A pharmaceutical composition comprising at least one of the compound of any one of claims 1 to 6, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof.

Use of a compound described in any one of claims 1 to 6, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof, or the drug composition described in claim 7 in the preparation of a drug, Preferably, the drug is a drug for diagnosing, preventing and/or treating a disease or condition mediated by p38α-MK2; Preferably, the drug is a p38α-MK2 inhibitor.

A method for diagnosing, preventing and/or treating a disease or condition mediated by p38α-MK2, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of at least one compound described in any one of claims 1 to 6, its racemate, stereoisomer, tautomer, isotopically labeled, nitrogen oxide, solvate, polymorph, pharmaceutically acceptable salt or prodrug thereof, or the drug composition described in claim 7, alone or, optionally, in combination with at least one other type of therapeutic agent.

The use as described in claim 8 or the method as described in claim 9, wherein the p38α-MK2-mediated disease or symptom is selected from ulcerative colitis, inflammatory enteritis, Crohn's disease, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, bone disease, osteoarthritis, septic shock, endotoxin shock, arthritis, sepsis, asthma, chronic obstructive pulmonary disease, cryopyrin-related periodic syndrome, rheumatoid arthritis, hidradenitis suppurativa, ankylosing spondylitis or cancer; preferably, the disease or symptom is selected from rheumatoid arthritis or ankylosing spondylitis.