Fused tricyclic compounds as kinase inhibitors

Abstract

A class of compounds containing tricyclic heteroaryl groups. Specifically, provided are compounds having a structure shown in the following formula (I), pharmaceutical compositions containing the compounds of formula (I), and the compounds, as well as isotopic derivatives, chiral isomers, allomers, different salts, prodrugs, preparations, etc. of these compounds. The compounds of formula (I) can effectively inhibit protein kinases (including EGFR, EGFR (C797S), ALK, and HPK1, etc.), thereby playing a role in treating various tumors.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry; in particular, the present invention relates to a novel class of derivatives containing tricyclic heteroaryl groups, a synthesis method thereof and an application thereof as one or more protein kinase inhibitors in the preparation of drugs for treating tumors and other related diseases. Background Art

[0002] Cancer, also known as malignant tumor, is one of the diseases with the highest incidence and mortality in the world. It is characterized by abnormal cell proliferation and metastasis, which spreads and metastasizes within a short time or a relatively short time after onset. Traditional treatment options include resection (if the conditions for resection are met), radiotherapy, and chemotherapy. Targeted therapy developed in recent years has the advantages of reducing toxicity and negative effects on patients, and improving survival rates. However, after using targeted drugs for a period of time, drug resistance will develop, and then the growth and spread of cancer cells will be abnormally rapid. Common cancers include: blood cancer, lung cancer, liver cancer, bladder cancer, rectal cancer, gastric cancer, etc.

[0003] Among all cancers, lung cancer ranks first in morbidity and mortality among all malignant tumors, among which non-small cell lung cancer (NSCLC) accounts for about 80% of all lung cancers.

[0004] Currently, the most effective means for non-small cell lung cancer is personalized targeted therapy, and common targets include C-met, ALK and EGFR.

[0005] Epidermal growth factor receptor tyrosine kinase (EGFR) is a transmembrane glycoprotein composed of 1186 amino acids and encoding a molecular weight of 170-kDa. EGFR can mediate multiple signal transduction pathways, transmit extracellular signals to cells, and play an important regulatory role in the proliferation, differentiation and apoptosis of normal cells and tumor cells. EGFR is a constitutively expressed component of many normal epithelial tissues (such as skin and hair follicles), while in most solid tumors, EGFR is overexpressed or highly expressed. For example, in lung cancer, the expression rate of EGFR reaches 40-80%. Therefore, selectively inhibiting EGFR and interfering with its mediated signal transduction pathway can achieve the purpose of treating lung cancer, opening up a feasible path for targeted treatment of lung cancer.

[0006] Anaplastic lymphoma kinase (ALK) is a transmembrane receptor tyrosine kinase that can mutate or fuse with other oncogenes in a variety of malignant tumors and is a tumor-causing driver gene. ALK inhibitors can be used to treat lung cancer.

[0007] Studies have shown that in patients with non-small cell lung cancer, especially after using epidermal growth factor receptor (EGFR) inhibitors for a period of time, EGFR mutations are most common, of which about 30% to 40% of Asian NSCLC patients carry EGFR mutations when diagnosed, especially middle-aged women with no smoking history. Therefore, developing a new generation of EGFR inhibitors to fight cancer mutations is a difficult problem that scientists have to overcome, and it is also one of the research hotspots in the field of biomedicine today.

[0008] There are three generations of EGFR inhibitors currently on the market, of which gefitinib is the first generation representative inhibitor. Erlotinib The second generation of EGFR-TKIs are afatinib and icotinib (Camena), the third generation of EGFR-TKIs are osimertinib (AZD9291). Osimertinib (AZD9291) is an oral small molecule third generation EGFR-TKI. It is the first lung cancer drug targeting EGFR T790M mutation. It can target EGFR gene mutations (including 18, 19, 21 mutations) and EGFR-TKI acquired resistance (T790M) in non-small cell lung cancer. Its advent has brought good survival benefits to more lung cancer patients. AZD9291 can significantly prolong survival by about one year. However, drug resistance develops very rapidly afterwards, generally within 9-13 months. In August 2016, the world's top academic journal Nature published a blockbuster article announcing a new generation of targeted drug compound EAI045 that can overcome AZD9291 resistance. It can be used for patients who are resistant to the first generation of drugs and have T790M mutations, or for patients who are resistant to AZD9291 and have C797S mutations. That is, the fourth-generation inhibitor is mainly aimed at patients with resistance mutations such as L858R / T790M double mutations and C797S mutations.

[0009] Hematopoietic progenitor kinase 1 (HPK1, also known as MAP4K1) is a member of the germinal center kinase family of serine / threonine kinases. It is mainly expressed by hematopoietic cells and is an intracellular negative regulator of T cell proliferation and signal transduction. It plays an important role in dendritic cell activation and is a new anti-cancer immunotherapy target. Therefore, inhibiting HPK1 with small molecule inhibitors has the potential to treat cancer and other diseases either as a single agent or in combination with other drugs. Summary of the invention

[0010] The purpose of the present invention is to provide a new type of protein kinase inhibitors.

[0011] In the first aspect of the present invention, there is provided a compound having a structure as shown in the following formula (I), or an optical isomer (including a racemate, a single enantiomer, and possible diastereomers), a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvate thereof:

[0012]

[0013] In formula (I):

[0014] “*” indicates chiral center;

[0015] Each R is independently C 1-4 alkyl;

[0016] Each R 1 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, OR h , or CN;

[0017] Each R 2 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, or CN;

[0018] Each R 3 Each is independently hydrogen, deuterium, or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atoms to which it is attached may optionally together form a carbonyl group (C=O);

[0019] Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, OR h , SR h NR h R h 、CN、C(O)R e 、C(O)OR h , C(O)NR h R h 、OC(O)R e NR h C(O)Re , or S(O) 2 R e ;

[0020] J and G are each independently NR f 、O、S、S(O)、S(O) 2 or CR g R g ;

[0021] m is 0, 1, 2, 3, or 4;

[0022] n is 0, 1, 2, or 3;

[0023] p is 0, 1, or 2;

[0024] q is 0, 1, 2, or 3;

[0025] R f For hydrogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O) 2 R e , or S(O) 2 NR h R h ; wherein each of the above groups is unsubstituted or substituted with 1-3 R e replace;

[0026] Each R e Each is independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Halogenated alkenyl, C 1-4 Alkoxy substituted C 2-4 Alkenyl, hydroxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkenyl, C 3-8 Cycloalkyl substituted C 2-4 alkenyl, 3- to 8-membered heterocyclic substituted C 2-4 Alkenyl, aryl substituted C 2-4 Alkenyl, heteroaryl substituted C2-4 Alkenyl, C 2-4 Alkynyl, C 2-4 Haloalkynyl, C 1-4 Alkoxy substituted C 2-4 Alkynyl, hydroxy substituted C 2-4 Alkynyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkynyl, C 3-8 Cycloalkyl substituted C 2-4 Alkynyl, 3- to 8-membered heterocyclic substituted C 2-4 Alkynyl, aryl substituted C 2-4 Alkynyl, heteroaryl substituted C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, or heteroaryl;

[0027] Each R g are independently selected from the group consisting of hydrogen, halogen, or C 1-4 Alkyl; or two R g The carbon atom connected to it together forms a carbonyl group (C=O); or two R g Together with the carbon atom to which it is attached, it forms a 3- to 8-membered ring structure, which optionally contains 0, 1 or 2 heteroatoms selected from N, O, S;

[0028] Each R h are independently hydrogen, or C 1-4 Alkyl; or two R h Together with the nitrogen atom to which it is attached, it forms a 3- to 8-membered heterocyclic group containing 1 or 2 N atoms and 0 or 1 heteroatom selected from O and S;

[0029] wherein each of the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO 2 , OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h NR h C(O)R e, or S(O) 2 R e , provided that the chemical structure formed is stable and meaningful; e and R h is defined as above;

[0030] Unless otherwise specified, the above-mentioned aryl group is an aromatic group containing 6 to 12 carbon atoms; the heteroaryl group is a 5- to 15-membered heteroaromatic group; and the cyclic structure is a saturated or unsaturated cyclic group containing or not containing heteroatoms.

[0031] In another preferred embodiment, formula (I) is:

[0032]

[0033] wherein each group is as defined above.

[0034] In another preferred embodiment, each R is independently C 1-2 alkyl;

[0035] Each R 1 are independently hydrogen, deuterium, halogen, or C 1-2 alkyl;

[0036] Each R 2 are independently hydrogen, deuterium, halogen, or C 1-2 alkyl;

[0037] Each R 3 Each is independently hydrogen or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atom to which it is attached together forms a carbonyl group (C=O);

[0038] Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, NR h R h , or NR h C(O)R e ;

[0039] m is 0, 1, or 2;

[0040] n is 0, 1, or 2;

[0041] p is 0, 1, or 2;

[0042] q is 0, 1, or 2;

[0043] Where R e and R h The definition of is as above.

[0044] In another preferred embodiment, formula (I) is:

[0045]

[0046] Where R 2 is F, Cl, or Br; each R 3 Each is independently hydrogen or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atom connected to it together forms a carbonyl group (C=O); each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, NR h R h , or NR h C(O)R e ; n is 0, 1, or 2; q is 0, 1, or 2; wherein J, G, and R e , and R h The definition of is as described above.

[0047] In another preferred embodiment, the structural fragment in formula (IIIa) Selected from:

[0048]

[0049] Represents the connection site between the above structural fragment and other structures in formula (IIIa);

[0050] Among them, each R 3 Each is independently hydrogen or C 1-4 Alkyl; when two R 3 When attached to the same carbon atom, the two R 3 and the carbon atoms connecting them can together form C=O;

[0051] Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ;

[0052] n is 0, 1, or 2; q is 0 or 1;

[0053] R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O) 2 R e , or S(O) 2 NR h R h wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO 2 , OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h NR h C(O)R e 、S(O) 2 R e , or S(O) 2 NR h R h ; where R e and R h The definition of is as above.

[0054] In another preferred embodiment, formula (I) is:

[0055]

[0056] Among them, each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ;q is 0 or 1;

[0057] R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O) 2 R e , or S(O) 2 NR h R h wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO 2 , OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h NR h C(O)R e 、S(O) 2 R e , or S(O) 2 NR h R h ; where R e and R h The definition of is as above.

[0058] In another preferred embodiment, the R f Selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O) 2 R e wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO2 , OR e , SR e NR e R e 、C(O)R e 、C(O)OR e 、C(O)NR e R e NR e C(O)R e 、S(O) 2 R e , or S(O) 2 NR h R h ; where R e and R h The definition of is as above.

[0059] In another preferred embodiment, formula (I) is:

[0060]

[0061] Among them, R 4 For hydrogen, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ;

[0062] R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O) 2 R e wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO 2 , OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h NR h C(O)R e、S(O) 2 R e , or S(O) 2 NR h R h ; where R e and R h The definition of is as above.

[0063] In another preferred embodiment, formula (I) is:

[0064]

[0065] Among them, R 4 For hydrogen, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ;

[0066] R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O) 2 R e wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO 2 , OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h NR h C(O)R e 、S(O) 2 R e , or S(O) 2 NR h R h ; where R e and R h is as defined in the first aspect of the present invention.

[0067] In another preferred embodiment, formula (I) is:

[0068]

[0069] Among them, R 4 For hydrogen, halogen, C 1-2 alkyl;

[0070] s and t are each independently 1, 2, or 3;

[0071] A is NR k , O, or CR g R g ; where R k For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, hydroxy substituted C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, di(C 1-4 Alkyl)amino-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O) 2 R e , or S(O) 2 NR h R h ; each R e are independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 1-4 Alkoxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, or heteroaryl; R g and R h is as defined in the first aspect of the present invention.

[0072] In another preferred embodiment, each of the R e Each independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, C 2-4 Alkenyl, C 1-4 Alkoxy substituted C 2-4Alkenyl, hydroxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 alkenyl, 3- to 6-membered heterocyclic substituted C 2-4 Alkenyl, aryl substituted C 2-4 Alkenyl, heteroaryl substituted C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, or heteroaryl.

[0073] In another preferred embodiment, each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, or NHC(O)CH=CH 2 .

[0074] In another preferred embodiment, the compound is selected from the following group:

[0075]

[0076]

[0077]

[0078]

[0079]

[0080] Wherein, “*” indicates a chiral center. Unless otherwise indicated as R or S, a compound with “*” indicates that it may be a racemate, an R configuration, or an S configuration.

[0081] In another preferred embodiment, used for:

[0082] (a) preparing a drug for treating a disease associated with protein kinase activity or expression;

[0083] (b) preparing protein kinase targeted inhibitors; and / or

[0084] (c) non-therapeutic inhibition of protein kinase activity in vitro;

[0085] Wherein, the protein kinase is selected from the following group: EGFR, EGFR (C797S), ALK, HPK1, etc., or a combination thereof.

[0086] The second aspect of the present invention provides a pharmaceutical composition, which comprises: (i) an effective amount of a compound of formula I as described in the first aspect of the present invention, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvate thereof; and (ii) a pharmaceutically acceptable carrier.

[0087] The third aspect of the present invention provides a method for preparing a compound of formula (I), characterized in that it comprises the steps of:

[0088]

[0089] Reacting a compound of formula 4-D1 with a compound of formula 1-A2 to obtain a compound of formula 4-D2-1 or 4-D2-2;

[0090] In the presence of a palladium catalyst, a compound of formula 4-D2-1 or 4-D2-2 is reacted with Me 4 Sn is reacted to obtain a compound of formula 4-D3-1 or formula 4-D3-2;

[0091] Performing a reduction reaction with a compound of formula 4-D3-1 or formula 4-D3-2 to obtain a compound of formula 4-D4-1 or formula 4-D4-2;

[0092] The compound of formula Ib is reacted with the compound of formula 4-D4-1 or the compound of formula 4-D4-2 to obtain the compound of formula IIIf or IIIg. Compound IIIf or compound IIIg is a part of the compound of formula (I).

[0093] The fourth aspect of the present invention provides a method for preparing a compound of formula (I), characterized in that it comprises the steps of:

[0094]

[0095] Reacting a compound of formula 5-E2 with a compound of formula Ib to obtain a compound of formula IIIh;

[0096] The compound of formula IIIh is subjected to a reductive amination reaction with the compound of formula 5-E3 to obtain the compound of formula IIIi. Compound IIIi is a part of the compound of formula (I).

[0097] The fifth aspect of the present invention provides a method for inhibiting the activity of EGFR and / or EGFR (C797S) and / or ALK and / or HPK1, which comprises the steps of: administering to the inhibited object an inhibitory effective amount of a compound of formula (I) as described in the first aspect of the present invention, or an optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate thereof, or administering to the inhibited object an inhibitory effective amount of a pharmaceutical composition as described in the third aspect of the present invention.

[0098] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0099] After long-term and in-depth research, the inventors unexpectedly discovered a class of protein kinase inhibitors containing tricyclic aromatic compounds with novel structures, as well as their preparation methods and applications. The protein kinase inhibitors can inhibit protein kinases such as EGFR, EGFR (C797S), ALK, and HPK1, and the compounds of the present invention can be used to treat various diseases related to the activity of EGFR (including various mutations produced), ALK, and HPK1. Among them, the inhibition of EGFR (C797S) is characterized by being able to overcome the drug resistance produced by the third-generation EGFR inhibitors. Specifically, the present invention includes a class of EGFR (especially EGFR (C797S)) inhibitors, which can effectively inhibit EGFR with L858R / T790M double mutation and C797S mutation. In addition, the compounds of the present invention can inhibit the immune target HPK1, and can be used alone or in combination with other drugs to treat a variety of cancers and other diseases. Based on the above findings, the inventors have completed the present invention.

[0100] the term

[0101] Unless otherwise specified, "or" mentioned in this document has the same meaning as "and / or" (referring to "or" and "and").

[0102] Unless otherwise specified, in all compounds of the present invention, each chiral carbon atom (chiral center) may be optionally in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

[0103] As used herein, the term "alkyl" when used alone or as part of another substituent refers to a straight chain (i.e., unbranched) or branched saturated hydrocarbon group containing only carbon atoms, or a combination of straight and branched chains. 1-10 ), it means that the alkyl group contains 1 to 10 carbon atoms. 1-8 The alkyl group refers to an alkyl group having 1 to 8 carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

[0104] As used herein, the term "alkenyl" when used alone or as part of another substituent refers to a straight or branched carbon chain group having at least one carbon-carbon double bond. The alkenyl group may be substituted or unsubstituted. When the alkenyl group is preceded by a carbon atom number limit (e.g., C 2-8), it means that the alkenyl group contains 2 to 8 carbon atoms. 2-8 The alkenyl group refers to an alkenyl group containing 2 to 8 carbon atoms, including ethenyl, propenyl, 1,2-butenyl, 2,3-butenyl, butadienyl, or the like.

[0105] As used herein, the term "alkynyl" when used alone or as part of another substituent refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond. The alkynyl group may be straight chain or branched, or a combination thereof. When the alkynyl group is preceded by a carbon atom number limit (e.g., C 2-8 When the term "alkynyl" is used, it means that the alkynyl contains 2 to 8 carbon atoms. 2-8 The term "alkynyl" refers to a straight or branched chain alkynyl group having 2 to 8 carbon atoms, including ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, sec-butynyl, tert-butynyl, or the like.

[0106] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated unit ring, bicyclic or polycyclic (fused, bridged or spiro) ring system group when used alone or as part of another substituent. 3-10 ) refers to the cycloalkyl group containing 3 to 10 carbon atoms. 3-8 "Cycloalkyl" refers to a saturated or partially saturated monocyclic or bicyclic alkyl group having 3-8 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or similar groups. "Spirocycloalkyl" refers to a bicyclic or polycyclic group in which monocyclic rings share a carbon atom (called a spiro atom), which may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. "Fused cycloalkyl" refers to a full-carbon bicyclic or polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. "Bridged cycloalkyl" refers to a full-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. The atoms contained in the cycloalkyl group are all carbon atoms. The following are some examples of cycloalkyl groups, and the present invention is not limited to the following cycloalkyl groups.

[0107]

[0108] Unless otherwise stated, the following terms used in the specification and claims have the following meanings. "Aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a conjugated π electron system, such as phenyl and naphthyl. The aryl ring can be fused to other cyclic groups (including saturated and unsaturated rings), but cannot contain heteroatoms such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be on a carbon atom on the ring with a conjugated π electron system. Aryl groups can be substituted or unsubstituted. The following are some examples of aryl groups, and the present invention is not limited to the aryl groups described below.

[0109]

[0110] "Heteroaryl" refers to a monocyclic or polycyclic group with aromaticity containing one to multiple heteroatoms (optionally selected from nitrogen, oxygen and sulfur), or a polycyclic group formed by condensing a heterocyclic group (containing one to multiple heteroatoms selected from nitrogen, oxygen and sulfur) with an aryl group, and the connection site is located on the aryl group. The heteroaryl group can be optionally substituted or unsubstituted. The following are some examples of heteroaryl groups, and the present invention is not limited to the following heteroaryl groups.

[0111]

[0112] "Heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl. Polycyclic heterocyclyls refer to heterocyclyls including spiro, fused rings and bridged rings. "Spirocyclic heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares an atom (called a spiro atom) with other rings in the system, wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. "Fused ring heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, and wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. "Bridged heterocyclic group" refers to a polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected. These may contain one or more double bonds, but none of the rings has a completely conjugated π-electron system, and one or more of the ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. If there are both saturated rings and aromatic rings in the heterocyclic group (for example, a saturated ring and an aromatic ring are fused together), the point of connection to the parent must be on the saturated ring. Note: When the point of connection to the parent is on the aromatic ring, it is called a heteroaryl group, not a heterocyclic group. The following are some examples of heterocyclic groups, and the present invention is not limited to the following heterocyclic groups.

[0113]

[0114] As used herein, the term "halogen" by itself or as part of another substituent refers to F, Cl, Br and I.

[0115] As used herein, the term "substituted" (with or without the "arbitrarily" modifier) ​​refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent is a substituent described accordingly in the preceding text, or a substituent appearing in the examples. Unless otherwise specified, an arbitrarily substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. A cyclic substituent, such as a heterocyclic group, may be connected to another ring, such as a cycloalkyl group, to form a spirobicyclic system, i.e., two rings have a common carbon atom. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. The substituents are, for example (but not limited to): C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group.

[0116] For the sake of convenience and in accordance with common understanding, the term "arbitrary substitution" or "optionally substituted" only applies to sites that can be substituted by substituents, and does not include those substitutions that are chemically unfeasible.

[0117] As used herein, unless otherwise specified, the term "pharmaceutically acceptable salt" refers to a salt that is suitable for contact with the tissue of a subject (e.g., a human) without producing undue side effects. In some embodiments, a pharmaceutically acceptable salt of a compound of the present invention includes a salt of a compound of the present invention having an acidic group (e.g., potassium salt, sodium salt, magnesium salt, calcium salt) or a salt of a compound of the present invention having a basic group (e.g., sulfate, hydrochloride, phosphate, nitrate, carbonate).

[0118] use:

[0119] The present invention provides a class of compounds of formula (I), or their deuterated derivatives, their salts, isomers (enantiomers or diastereomers, if any), prodrugs, hydrates, solvates, pharmaceutically acceptable carriers or excipients for use in inhibiting protein kinases. The protein kinases referred to herein include EGFR, EGFR (C797S), ALK, and HPK1, but are not limited to the above kinases.

[0120] The compounds of the present invention can be used as one or more kinase inhibitors. For example, in some embodiments, a certain class of compounds of the present invention can be used as EGFR and / or EGFR(C797S)) and / or ALK and / or HPK1 kinase inhibitors.

[0121] In cancer patients, the expression or activity of the various protein kinases mentioned above are significantly increased. These overexpressed and / or abnormal protein kinase activity levels are directly related to the occurrence and development of tumors. The compounds of the present invention are single and / or dual inhibitors of these protein kinases. Diseases can be prevented, alleviated or cured by regulating the activity of these protein kinases. The diseases referred to include liver cancer, colorectal cancer, bladder cancer, pharyngeal cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, pancreatic cancer, colon cancer, skin cancer, lymphoma, gastric cancer, multiple myeloma and solid tumors, etc.

[0122] In a sense, dual protein kinase inhibitors interfere with two different kinases at the same time, and the anti-tumor effects they produce are often additive, so they have the potential to more effectively treat various cancers.

[0123] The compounds of the present invention can be used in combination with biological agents such as PD-1 inhibitors and As a combination drug to treat various cancers and related diseases.

[0124] The compounds of the present invention and their deuterated derivatives, as well as pharmaceutically acceptable salts or isomers thereof (if present) or hydrates thereof and / or compositions can be formulated with pharmaceutically acceptable excipients or carriers, and the resulting compositions can be administered to mammals, such as men, women and animals, in vivo for the treatment of conditions, symptoms and diseases. The compositions can be tablets, pills, suspensions, solutions, emulsions, capsules, aerosols, sterile injections, sterile powders, etc. In some embodiments, pharmaceutically acceptable excipients include microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, mannitol, hydroxypropyl-β-cyclodextrin, β-cyclodextrin (increased), glycine, disintegrants (such as starch, cross-linked sodium carboxymethyl cellulose, complex silicates and high molecular weight polyethylene glycol), granulation binders (such as polyvinyl pyrrolidone, sucrose, gelatin and gum arabic) and lubricants (such as magnesium stearate, glycerol and talc). In a preferred embodiment, the pharmaceutical composition is a dosage form suitable for oral administration, including but not limited to tablets, solutions, suspensions, capsules, granules, and powders. The amount of the compound of the present invention or the pharmaceutical composition administered to the patient is not fixed, and is usually administered in a pharmaceutically effective amount. At the same time, the amount of the compound actually administered can be determined by the physician according to actual conditions, including the disease to be treated, the selected route of administration, the actual compound administered, the individual conditions of the patient, etc. The dosage of the compound of the present invention depends on the specific use of the treatment, the mode of administration, the patient's state, and the physician's judgment. The ratio or concentration of the compound of the present invention in the pharmaceutical composition depends on various factors, including dosage, physicochemical properties, route of administration, etc.

[0125] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form new or preferred technical solutions.

[0126] General synthesis method of compounds

[0127] The compound of formula I of the present invention can be prepared by the following method:

[0128]

[0129] In an inert solvent, compound (Ia) is reacted with compound (Ib) to obtain compound (I);

[0130] In the above formulae, the definitions of the groups are as described above. The reagents and conditions of each step can be selected from conventional reagents or conditions for such preparation methods in the art. After the structure of the compound of the present invention is disclosed, the above selection can be made by those skilled in the art based on the knowledge in the art.

[0131] More specifically, the compound of general formula I of the present invention can be prepared by the following method, however, the conditions of the method, such as reactants, solvents, bases, the amount of the compounds used, reaction temperature, reaction time, etc. are not limited to the following explanation. The compound of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combination can be easily performed by a person skilled in the art to which the present invention belongs.

[0132] In the preparation method of the present invention, each reaction is usually carried out in an inert solvent at a reaction temperature of usually -20 to 150°C (preferably 0 to 120°C). The reaction time of each step is usually 0.5 to 48 hours, preferably 2 to 12 hours.

[0133] Reaction Scheme 1 describes the general synthesis of intermediates 1-A5-1 and 1-A5-2:

[0134] Reaction 1:

[0135]

[0136] Reaction 2 describes the general synthesis of intermediates 2-B3-1 and 2-B3-2:

[0137] Reaction 2:

[0138]

[0139] Reaction 3 describes the general synthesis of intermediate 3-C7:

[0140] Reaction 3:

[0141]

[0142] Compound IIIa is a part of compound I. Reaction Scheme 4 describes the general synthesis method of compound IIIa: Reaction Scheme 4:

[0143]

[0144] Compound IIIb is a part of compound I. Reaction Scheme 5 describes the general synthesis method of compound IIIb: Reaction Scheme 5:

[0145]

[0146] Compound IIIc is a part of compound I. Reaction Scheme 6 describes the general synthesis method of compound IIIc: Reaction Scheme 6:

[0147]

[0148] Compound IIId is a part of compound I. Reaction Scheme 7 describes the general synthesis method of compound IIId: Reaction Scheme 7:

[0149]

[0150] Compound IIIe is a part of compound I. Reaction formula 8 describes the general synthesis method of compound IIIe: Reaction formula 8:

[0151]

[0152] Compounds IIIf and IIIg are part of compound I. Reaction Scheme 9 describes the general synthesis method of compounds IIIf and IIIg:

[0153] Reaction 9:

[0154]

[0155] Compounds IIIh and IIIi are part of compound I. Reaction Scheme 10 describes the general synthesis method of compounds IIIh and IIIi:

[0156] Reaction 10:

[0157]

[0158] In the above reaction formula 1-10, R and R 1 , R 2 , R 4 , R f The definitions of , m, p, q, s, t, and A are the same as those described above.

[0159] Pharmaceutical compositions and methods of administration

[0160] Since the compounds of the present invention have excellent inhibitory activity against a series of protein kinases, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate diseases related to the activity or expression of protein kinases such as EGFR, EGFR (C797S), ALK, and HPK1.

[0161] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 5-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0162] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0163] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0164] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0165] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0166] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0167] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0168] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0169] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0170] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0171] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0172] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of a skilled physician.

[0173] The main advantages of the present invention include:

[0174] 1. Provided is a compound as shown in formula (I).

[0175] 2. Provided are a novel structured inhibitor of the activity of protein kinases such as EGFR, EGFR (C797S), ALK, and HPK1, as well as its preparation and use. The inhibitor can inhibit the activity of the above protein kinases at extremely low concentrations.

[0176] 3. Provided is a class of pharmaceutical compositions for treating diseases associated with the activity of protein kinases such as EGFR, EGFR(C797S), ALK, and HPK1.

[0177] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0178] Example 1: Preparation of Compound 1

[0179]

[0180] KOH (1.01 g, 18.0 mmoL) was added to a dry DMSO (15 mL) solution of 1,2-difluoro-4-nitrobenzene (1a, 1.11 g, 7.0 mmoL) and racemic tert-butyl 3-hydroxymethylpiperazine-1-carbonate (1b, 1.30 g, 6.0 mmoL). The mixture was stirred at room temperature for 3 hours and then heated to 60°C for 8 hours. After the reaction was completed, ice water was added to the reaction solution. The solid formed was collected by filtration, washed with water, and then purified by flash column chromatography (petroleum ether: ethyl acetate = 6:1) to obtain a yellow solid product 1c (1.41 g, yield 70.0%). 1 H NMR (600 MHz, CDCl 3 ): δ7.79(dd,J=9.1Hz,2.6Hz,1H),7.66(d,J=2.6Hz,1H),6.76(d,J=9.2Hz,1H),4.30(dd,J=11.0Hz,3.0Hz,1H),4.18-4.09(m,2H),3 .99(dd,J=11.1Hz,8.0Hz,1H),3.79(d,J=11.0Hz,1H),3.35-3.31(m,1H),3.04-2.93(m,2H),2.67(br,1H),1.49(s,9H); HRMS(+):m / z 358.1370([M+Na] + ,C 16 H 21 N 3 O 5 Na +calcd:358.1373.

[0181] Compound 1c (2.0 g, 6.0 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (5 mL) was added under stirring at room temperature. After stirring at room temperature for 1 hour, the reaction was completed by TLC monitoring. The reaction solution was concentrated under reduced pressure to remove trifluoroacetic acid, and the residue was dissolved in dichloromethane (30 mL), and the pH was adjusted to 9-10 with aqueous sodium carbonate solution (1 M), and the organic phase and the aqueous phase were separated. The aqueous layer was extracted twice with dichloromethane. The combined organic phase was washed once with saturated brine (30 mL) and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure to obtain yellow solid compound 1d (1.2 g). MS m / z 236.1[M+H] + .

[0182] The above compound 1d (1.2 g) was dissolved in methanol (20 mL), and a formaldehyde aqueous solution (37%, 6 mL) was added, followed by 2 drops of acetic acid. After stirring at room temperature for 30 minutes, sodium cyanoborohydride (0.8 g, 12.7 mmol) was added, and then stirring was continued at room temperature for 3 hours. TLC monitored the completion of the reaction. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (DCM: MeOH = 60: 1) to obtain a yellow solid compound 1e (1.0 g). MS m / z 250.2 [M+H] + .

[0183] Compound 1e (145 mg, 0.58 mmol) and palladium carbon catalyst (10%, 15 mg) were added to methanol (3 mL) at room temperature. The reaction mixture was reacted for 1 hour at room temperature and 1 atmosphere of hydrogen atmosphere. The reaction was completed by TLC monitoring. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain brown solid compound 1f (100 mg). The crude product was used directly in the next step. MS m / z 220.2 [M+H] + .

[0184] Compound 1g (3.0 g, 14 mmol) and compound 1h (1.2 g, 15 mmol) were dissolved in DMF (60 mL), and Pd(OAc) was added successively. 2 (0.2g,) 、 X-Phos (0.3 g), and K 3 PO 4 (5.9 g, 28 mmol). The mixture was replaced with nitrogen three times, and then heated to 120°C under a nitrogen atmosphere until the reaction was complete. The mixture was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (DCM:MeOH=1:40) to obtain a white solid compound 1i (0.75 g, yield 33%).

[0185] To a solution of compound 1i (750 mg, 4.4 mmol) and compound 1j (812 mg, 4.4 mmol) in IPA (20 mL) was added K 2 CO 3 (1.2 g, 8.8 mmol). The resulting suspension was heated to 105°C in a microwave reactor and reacted for 1 h. The mixture was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (ethyl acetate: petroleum ether = 1:1) to obtain a milky white solid compound 1k (450 mg, yield 32%). Liquid chromatography conditions: chromatographic column X Brdige C18: 4.6 mm*30 mm*3.5 um, mobile phase A: water (0.01 mol / L NH 4 HCO 3 ), mobile phase B: acetonitrile, flow rate 2.0 mL / min, gradient elution from 10% B to 95% B in 0.5 min, then constant at 95% B for 1.5 min. Purity 95%, retention time 0.95 min; MS m / z 316.0 [M+H] + .

[0186] Compound 1f (22 mg, 0.1 mmol) and compound 1k (31.5 mg, 0.1 mmol) were heated to 80°C in a hydrochloric acid aqueous solution (2 Min EtOH, 2 mL) and stirred until the reaction was complete. The reaction mixture was concentrated under reduced pressure, and the residue was separated by preparative HPLC to obtain a light yellow solid compound 1 (5.6 mg, yield 11%). Liquid chromatography conditions: chromatographic column X BrdigeC18: 4.6 mm*30 mm*3.5 um, mobile phase A: water (0.01 mol / L NH 4 HCO 3 ), mobile phase B: acetonitrile, flow rate 2.0 mL / min, gradient elution from 5% B to 100% B for 1.6 minutes, then constant 100% B for 1.4 minutes. Purity 90%. Retention time 1.56 min. 1 H NMR (400MHz, CD 3OD)δ8.48-8.52(m,1H),8.03(s,1H),7.53-7.61(m,2H),7.24-7.26(m,1H),7.03-7.04 (m,1H),6.89-6.91(m,1H),6.75-6.77(m,1H),4.21-4.24(m,1H),3.94-3.99(m,1H),3 .72-3.75(m,1H),3.08-3.09(m,1H),2.97-3.00(m,1H),2.87-2.90(m,1H),2.72-2.73 (m,1H),2.38(s,3H),2.29-2.36(m,1H),1.87-1.92(m,1H),1.86(s,3H),1.84(s,3H). MS m / z 499.2[M+H] + .

[0187] Example 2: Preparation of Compound 1R

[0188]

[0189] Compound 1R-1a (650 mg, 2.60 mmol) and palladium-carbon catalyst (10%, 65 mg) were added to methanol (15 mL) at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and hydrogen (1 atmosphere). TLC monitored the reaction completion. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product, which was then separated and purified by silica gel column chromatography (dichloromethane: methanol = 40: 1 to 20: 1) to obtain a yellow oily compound 1R-1b (180 mg, yield 32%). 1 H NMR (500 MHz, CDCl 3 )δ6.62(d,J=8.4Hz,1H),6.24(dd,J=8.4,2.6Hz,1H),6.21(d,J=2.5Hz,1H),4.14(d d,J=10.5,2.6Hz,1H),4.03-3.95(m,1H),3.56(dt,J=11.5,2.7Hz,1H),3.36(s,br., 2H),3.12-3.03(m,1H),2.91(dd,J=11.3,2.2Hz,1H),2.81-2.75(m,1H),2.72(td,J= 11.7, 3.1Hz, 1H), 2.33 (s, 3H), 2.25 (td, J = 11.5, 3.2Hz, 1H), 1.83 (t, J = 10.6Hz, 1H).

[0190] Compound 1R-1b (70 mg, 0.32 mmol) and compound 1k (72 mg, 0.23 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.24 mL) was added. The above reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials, stopped the reaction, and after cooling, the reaction solution was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 40: 1 to 20: 1) to obtain an off-white solid compound 1R (79 mg, yield 50%). 1 H NMR (500 MHz, CDCl 3 )δ10.88(s,1H),8.65(dd,J=8.4,4.3Hz,1H),8.06(s,1H),7.48(dd,J=8.4,7.4Hz,1H),7.29-7.22(m, 1H),7.11-7.06(m,2H),6.92(dd,J=8.6,2.5Hz,1H),6.74-6.70(m,2H),4.20(dd,J=10.5,2.7Hz,1H), 4.03(dd,J=10.4,9.1Hz,1H),3.67-3.61(m,1H),3.22-3.15(m,1H),2.94(d,J=10.9Hz,1H),2.84-2.7 7(m,2H),2.36(s,3H),2.26(td,J=11.6,3.2Hz,1H),1.87(d,J=10.7Hz,1H),1.84(s,3H),1.81(s,3H). MS m / z 499.4[M+H] + .

[0191] Example 3: Preparation of Compound 1S

[0192]

[0193] Compound 1S-1a (800 mg, 3.20 mmol) and palladium-carbon catalyst (10%, 80 mg) were added to methanol (10 mL) at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and under a hydrogen (1 atmosphere) atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 40: 1 to 20: 1) to obtain a yellow oily compound 1S-1b (400 mg, yield 57%). 1 H NMR (500 MHz, DMSO-d 6)δ6.54(d,J=8.4Hz,1H),6.06(d,J=8.5Hz,1H),6.01(s,1H),4.47(s,br.,2H),4.10(d,J=10.4Hz,1H),3.80(t,J=9.8Hz,1H),3.49(d,J=11.4 Hz,1H),2.82(t,J=9.5Hz,2H),2.72(d,J=10.6Hz,1H),2.48-2.40(m,1H),2.19(s,3H),2.07(td,J=11.0,1.9Hz,1H),1.66(t,J=10.5Hz,1H).

[0194] Compound 1S-1b (70 mg, 0.32 mmol) and compound 1k (72 mg, 0.23 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.24 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 40: 1 to 20: 1) to obtain an off-white solid compound 1S (44 mg, yield 38%). 1 H NMR (500 MHz, CDCl 3 )δ10.88(s,1H),8.65(dd,J=8.3,4.4Hz,1H),8.05(s,1H),7.48(dd,J=8.4,7.4Hz,1H),7.28-7.22(m, 1H),7.11-7.06(m,2H),6.92(dd,J=8.7,2.5Hz,1H),6.79-6.72(m,2H),4.20(dd,J=10.5,2.6Hz,1H), 4.03(dd,J=10.4,9.0Hz,1H),3.67-3.61(m,1H),3.22-3.15(m,1H),2.95(d,J=10.9Hz,1H),2.85-2.7 8(m,2H),2.36(s,3H),2.27(td,J=11.5,3.2Hz,1H),1.87(d,J=10.7Hz,1H),1.83(s,3H),1.81(s,3H). MS m / z 499.4[M+H] + .

[0195] Example 4: Preparation of Compound 2R

[0196]

[0197] Compound 2R-1a (63 mg, 0.20 mmol) and compound 1k (46 mg, 0.14 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.15 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 40: 1 to 20: 1) to obtain an off-white solid compound 2R (21 mg, yield 25%). 1 H NMR (500 MHz, CDCl 3 )δ10.91(s,1H),8.64(dd,J=8.3,4.4Hz,1H),8.06(s,1H),7.48(dd,J=8.4,7.4Hz,1H),7.30-7.23(m,1H),7.16(d,J=2.5Hz, 1H),7.13-7.09(m,1H),6.95(dd,J=8.7,2.5Hz,1H),6.80(s,1H),6.75(d,J=8.8Hz,1H),4.26(dd,J=10.7,2.7Hz,1H),4.04( dd,J=10.7,8.2Hz,1H),3.86-3.77(m,2H),3.75-3.70(m,1H),3.28-3.22(m,1H),3.10(td,J=11.6,2.8Hz,1H),2.90(td,J=1 1.9, 3.1Hz, 1H), 2.75 (t, J = 11.0Hz, 1H), 2.30-2.23 (m, 1H), 1.84 (s, 3H), 1.82 (s, 3H), 1.23-1.68 (m, 2H), 1.05-1.00 (m, 2H). MSm / z 589.4[M+H] + .

[0198] Example 5: Preparation of Compound 3R

[0199]

[0200] Compound 3R-1a (86 mg, 0.30 mmol) and compound 1k (67 mg, 0.20 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.22 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to give an off-white solid compound 3R (70 mg, yield 62%).1 H NMR (500 MHz, CDCl 3 )δ10.89(s,1H),8.66(dd,J=8.2,4.4Hz,1H),8.05(s,1H),7.48(dd,J=8.4,7.4Hz,1H),7.28-7.22(m,1H),7.11- 7.06(m,2H),6.92(dd,J=8.7,2.5Hz,1H),6.75-6.70(m,2H),4.20(dd,J=10.4,2.7Hz,1H),4.09-3.98(m,3H),3.6 7(d,J=11.5Hz,1H),3.39(t,J=11.4Hz,2H),3.17-3.10(m,1H),3.06(d,J=11.0,1H),2.91(d,J=10.4Hz,1H),2.77 (td,J=11.5,2.9Hz,1H),2.53-2.41(m,2H),2.03(t,J=10.5Hz,1H),1.84(s,3H),1.81(s,3H),1.80-1.76(m,4H). MS m / z 569.6[M+H] + .

[0201] Example 6: Preparation of Compound 4R

[0202]

[0203] Palladium-carbon catalyst (10%, 250 mg) was added to a methanol (35 mL) solution of compound 4R-1a (3.0 g, 8.94 mmol) at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and hydrogen (1 atmosphere). TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1 to 1:1) to obtain a dark purple solid compound 4R-1b (2.48 g, yield 91%). 1 H NMR (500 MHz, CDCl 3)δ6.63(d,J=8.5Hz,1H),6.25(dd,J=8.5,2.5Hz,1H),6.21(d,J=2.5Hz,1H),4.17(dd,J=10.6,2.6Hz,1H),4.14-4.00(m,2H),3.96(dd,J=10.5 ,9.1Hz,1H),3.56(d,J=11.0Hz,1H),3.38(s,2H),3.04(s,1H),2.95(ddd,J=11.7,5.5,2.7Hz,1H),2.59(td,J=11.8,3.1Hz,2H),1.47(s,9H).

[0204] Compound 4R-1b (100 mg, 0.33 mmol) and compound 1k (74 mg, 0.24 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.25 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated by preparative thin plate chromatography (dichloromethane: acetone = 1: 1, 2% triethylamine) to obtain an off-white solid compound 4R (6 mg, yield 5%). 1 H NMR (500MHz, CD 3 OD)δ8.49(dd,J=8.2,4.6Hz,1H),8.03(s,1H),7.63-7.52(m,2H),7.26-7.22(m,1H),7.08(d ,J=2.4Hz,1H),6.93(dd,J=8.7,2.5Hz,1H),6.80(d,J=8.9Hz,1H),4.26(dd,J=10.7,2.5Hz,1 H),4.00(dd,J=10.8,8.2Hz,1H),3.90(d,J=12.7Hz,1H),3.27-3.16(m,3H),3.11(td,J=12.5 ,3.5Hz,1H),2.81(td,J=12.6,3.1Hz,1H),2.75(t,J=11.4Hz,1H),1.86(s,3H),1.83(s,3H). MS m / z 485.4[M+H] + .

[0205] Example 7: Preparation of Compound 4S

[0206]

[0207] Under an ice-water bath, to a stirred solution of compound 4S-1a (500 mg, 2.10 mmol) in 1,4-dioxane (8 mL), add an aqueous NaOH solution (1 N, 2 mL), followed by di-tert-butyl dicarbonate (510 mg, 2.30 mmol). The reaction mixture was warmed to room temperature and stirred for about 30 min. TLC monitored the disappearance of the reaction raw materials. The reaction solution was poured into ice water and extracted with dichloromethane (20 mL x3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid crude product 4S-1b. The resulting crude product was directly used in the next step reaction.

[0208] At room temperature, palladium-carbon catalyst (10%, 80 mg) was added to a methanol (20 mL) solution of crude product 4S-1b. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and 1 atmosphere of hydrogen atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a white solid compound 4S-1c (432 mg, two-step total yield 67%). MS m / z 306.3 [M+H] + . Compound 4S-1c (48 mg, 0.16 mmol) and compound 1k (50 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (2 mL), and then HCl methanol solution (2.5 M, 0.16 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the disappearance of the reaction raw materials. The reaction was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to give an off-white solid compound 4S (45 mg, yield 57%). 1 H NMR (400 MHz, CDCl 3)δ10.86(s,1H),8.65(dd,J=8.4,4.5Hz,1H),8.05(s,1H),7.48(dd,J=8.4,7.4Hz,1H),7.31-7.21( m,1H),7.13-7.05(m,2H),6.92(dd,J=8.7,2.5Hz,1H),6.84(s,1H),6.72(d,J=8.8Hz,1H),4.18(dd ,J=10.5,2.5Hz,1H),4.01(dd,J=10.4,9.1Hz,1H),3.62(d,J=11.4Hz,1H),3.17(d,J=12.3Hz,1H), 3.09-2.93(m,3H),2.66(td,J=11.7,3.3Hz,1H),2.54(t,J=10.9Hz,1H),1.84(s,3H),1.81(s,3H). MS m / z485.3[M+H] + .

[0209] Example 8: Preparation of Compound 5R

[0210]

[0211] DIPEA (32 mg, 0.24 mmol) was added to a solution of compound 4R (60 mg, 0.12 mmol) in N,N-dimethylformamide (1.0 mL) at 0°C, followed by a solution of acetyl chloride (12 mg, 0.15 mmol) in dichloromethane (1.0 mL) was slowly added dropwise. After the addition was complete, the reaction solution was maintained at 0°C and stirred for 1 h. TLC monitored the completion of the reaction. Water (10 mL) was added to the reaction solution, and then extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: acetone = 2:1, 2% triethylamine) to give an off-white solid compound 5R (29 mg, yield 46%). 1 H NMR (500 MHz, CDCl 3)δ10.90(s,1H),8.65(dd,J=7.9,4.5Hz,1H),8.06(s,1H),7.48(t,J=8.0Hz,1H),7.30-7.23(m,1H),7.19- 7.08(m,2H),6.94(ddd,J=32.3,8.7,2.5Hz,1H),6.82(d,J=3.6Hz,1H),6.74(dd,J=14.9,8.8Hz,1H),4.75 -4.57(m,1H),4.26(dd,J=10.7,2.6Hz,1H),4.04(td,J=10.7,8.6Hz,1H),3.90(d,J=13.2Hz,1H),3.76-3. 67(m,1H),3.11-2.96(m,2H),2.75-2.64(m,1H),2.51-2.42(m,1H),2.16(S,3H),1.84(s,3H),1.81(s,3H). MS m / z 527.5[M+H] + .

[0212] Example 9: Preparation of Compound 6R

[0213]

[0214] DIPEA (32 mg, 0.24 mmol) was added to a solution of compound 4R (60 mg, 0.12 mmol) in N,N-dimethylformamide (1.0 mL) at 0°C, followed by a solution of acryloyl chloride (14 mg, 0.15 mmol) in dichloromethane (1.0 mL) which was then slowly added dropwise. After the addition was complete, the reaction solution was stirred for 1 h at 0°C. TLC monitored the completion of the reaction. Water (10 mL) was added to the reaction solution, which was then extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: acetone = 2:1, 2% triethylamine) to give an off-white solid compound 6R (21 mg, yield 32%). 1 H NMR (500 MHz, CDCl 3)δ10.94(s,1H),8.64(dd,J=8.4,4.4Hz,1H),8.05(s,1H),7.48(t,J=8.1Hz,1H),7.31-7.23(m,1H),7.21 -7.08(m,2H),7.01-6.82(m,2H),6.79-6.67(m,1H),6.67-6.55(m,1H),6.35(d,J=16.8Hz,1H),5.77(dd, J=10.6,1.7Hz,1H),4.72(dd,J=50.4,11.5Hz,1H),4.34-4.19(m,1H),4.13-3.87(m,1H),3.74(d,J=11.4 Hz,1H),3.47-3.37(m,1H),3.13-2.93(m,2H),2.73(m,1H),2.61-2.52(m,1H),1.84(s,3H),1.82(s,3H). MS m / z 539.5[M+H] + .

[0215] Example 10: Preparation of Compound 6S

[0216]

[0217] At 0°C, DIPEA (32 mg, 0.24 mmol) was added to a solution of compound 4S (60 mg, 0.12 mmol) in N,N-dimethylformamide (0.5 mL), followed by a solution of acryloyl chloride (14 mg, 0.15 mmol) in dichloromethane (2.0 mL) was slowly added dropwise. After the addition was complete, the reaction was stirred at 0°C for 1 h. TLC monitored the completion of the reaction. Water (10 mL) was added to the reaction solution, and then extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to give a yellow solid compound 6S (16 mg, yield 24%). 1 H NMR (400 MHz, CDCl 3)δ10.89(s,1H),8.64(dd,J=8.4,4.4Hz,1H),8.06(s,1H),7.48(t,J=7.9Hz,1H),7.31-7.22(m,1H),7.20-7 .07(m,2H),7.00-6.80(m,2H),6.79-6.69(m,1H),6.68-6.54(m,1H),6.34(dd,J=16.8,1.4Hz,1H),5.77(dd, J=10.5,1.8Hz,1H),4.71(dd,J=40.3,12.5Hz,1H),4.36-4.20(m,1H),4.13-3.88(m,1H),3.73(d,J=12.1Hz ,1H),3.48-3.34(m,1H),3.14-2.96(m,2H),2.79-2.66(m,1H),2.64-2.50(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 539.6[M+H] + .

[0218] Example 11: Preparation of Compound 7R

[0219]

[0220] To a solution of compound 4R (48.5 mg, 0.10 mmol) in N,N-dimethylformamide (0.5 mL), a solution of 2-bromoethanol (7R-1a, 6.3 mg, 0.05 mmol) in toluene (1.5 mL) was added. The reaction mixture was then placed in a sealed tube, heated to reflux temperature, and stirred for 1 h. The reaction was monitored by TLC. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to give a yellow solid compound 7R (13 mg, yield 50%). 1 H NMR (500 MHz, CDCl 3)δ10.89(s,1H),8.66(dd,J=8.5,4.4Hz,1H),8.05(s,1H),7.48(dd,J=8.4,7.6Hz,1H),7.29-7.20(m,1H),7.1 2-7.07(m,2H),6.92(dd,J=8.7,2.5Hz,1H),6.81(s,1H),6.72(d,J=8.8Hz,1H),4.20(dd,J=10.5,2.6Hz,1H),4 .03(dd,J=10.4,9.1Hz,1H),3.70-3.63(m,3H),3.19-3.11(m,1H),3.02(d,J=10.9,1H),2.90-2.85(m,1H),2. 83-2.74(m,1H),2.66-2.56(m,2H),2.40(td,J=11.5,3.1Hz,1H),2.04-1.95(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 529.5[M+H] + .

[0221] Example 12: Preparation of Compound 8R

[0222]

[0223] Potassium carbonate (27.6 mg, 0.20 mmol) was added to N, N-dimethylformamide (0.5 mL) of compound 4R (48.5 mg, 0.10 mmol), followed by dropwise addition of a toluene (1.5 mL) solution of ethyl bromide (8R-1a, 10.9 mg, 0.10 mmol). The reaction mixture was then placed in a sealed tube, heated to reflux temperature, and stirred for 3 h. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature, water (10 mL) was added, and then extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin plate chromatography (dichloromethane: acetone = 2: 1, 2% triethylamine) to give a white solid compound 8R (18 mg, yield 35%). 1 H NMR (500 MHz, CDCl 3)δ10.88(s,1H),8.65(dd,J=8.5,4.5Hz,1H),8.05(s,1H),7.48(t,J=7.9Hz,1H),7.29-7.21(m,1H),7.12-7.0 4(m,2H),6.92(dd,J=8.7,2.3Hz,1H),6.78(s,1H),6.72(d,J=8.7Hz,1H),4.21(d,J=10.4,1H),4.03(t,J=9.7 Hz,1H),3.65(d,J=11.8Hz,1H),3.18(t,J=9.7Hz,1H),3.05(d,J=11.4Hz,1H),2.94-2.87(m,1H),2.85-2.75( m,1H),2.53-2.45(m,2H),2.24(td,J=11.4,2.6Hz,1H),1.85-1.82(s,3H),1.81(s,3H),1.14(t,J=7.2Hz,3H). MS m / z513.3[M+H] + .

[0224] Example 13: Preparation of Compound 9R

[0225]

[0226] Potassium carbonate (27.6 mg, 0.20 mmol) was added to a solution of compound 4R (48.5 mg, 0.10 mmol) in N,N-dimethylformamide (0.6 mL), and then a solution of bromoacetonitrile (9R-1a, 13.2 mg, 0.11 mmol) in acetonitrile (1.5 mL) was added dropwise. The reaction mixture was stirred at room temperature overnight. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to give an off-white solid compound 9R (26 mg, yield 50%). 1 H NMR (500 MHz, CDCl 3)δ10.89(s,1H),8.65(dd,J=8.3,4.4Hz,1H),8.06(s,1H),7.51-7.46(m,1H),7.29-7.23(m,1H),7.16-7 .08(m,2H),6.94(dd,J=8.7,2.5Hz,1H),6.79(s,1H),6.73(d,J=8.8Hz,1H),4.21(dd,J=10.5,2.7Hz,1H ),4.06(dd,J=10.5,9.0Hz,1H),3.74-3.69(m,1H),3.60(s,2H),3.23-3.17(m,1H),2.90(dd,J=10.4,2. 0Hz,1H),2.85-2.74(m,2H),2.68(td,J=11.1,3.2Hz,1H),2.30-2.23(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 524.4[M+H] + .

[0227] Example 14: Preparation of Compound 10R

[0228]

[0229] Potassium carbonate (27.6 mg, 0.20 mmol) was added to a solution of compound 4R (48.5 mg, 0.10 mmol) in N,N-dimethylformamide (0.6 mL), and then a solution of 2-chloroethyl methyl ether (10R-1a, 9.5 mg, 0.10 mmol) in toluene (1.5 mL) was slowly added dropwise. After the addition was completed, the reaction mixture was placed in a sealed tube, heated to 90 ° C, and stirred for 2 h. TLC monitoring showed that there was still a lot of raw material remaining in the reaction, and DIPEA (25.9 mg, 0.20 mmol) was added and the reaction was continued for 2 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated by preparative thin plate chromatography (dichloromethane: acetone = 2:1, 2% triethylamine) to obtain a white solid compound 10R (12 mg, yield 22%). 1 H NMR (500 MHz, CDCl 3)δ10.89(s,1H),8.65(dd,J=8.3,4.3Hz,1H),8.05(s,1H),7.47(dd,J=8.3,7.4Hz,1H),7.29-7.22(m,1H),7.12-7.07(m,2H ),6.91(dd,J=8.7,2.5Hz,1H),6.80(s,1H),6.72(d,J=8.8Hz,1H),4.20(dd,J=10.5,2.6Hz,1H),4.02(dd,J=10.5,8.8Hz,1 H),3.64(d,J=11.7Hz,1H),3.59(t,J=5.3Hz,2H),3.38(s,3H),3.28(dd,J=12.8,7.3Hz,1H),3.10(d,J=10.8Hz,1H),2.98( d,J=10.7Hz,1H),2.94-2.86(m,1H),2.68(d,J=3.3Hz,2H),2.42-2.31(m,1H),2.04-1.92(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 543.4[M+H] + .

[0230] Example 15: Preparation of Compound 11R

[0231]

[0232] To a solution of compound 4R (60 mg, 0.12 mmol) in dichloromethane (1.5 mL), N-methyl-4-piperidone (11R-1a, 42 mg, 0.37 mmol) and 2 drops of acetic acid were added in sequence. The reaction mixture was stirred at room temperature for 1 h. It was then cooled to 0 ° C and sodium triacetoxyborohydride (80 mg, 0.37 mmol) was added to the mixture in batches. The reaction was stirred overnight at room temperature. TLC monitored the completion of the reaction. Water was added to the reaction solution to quench the reaction, and then a saturated sodium bicarbonate solution was added, and then extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to give a white solid compound 11R (9 mg, yield 13%). 1 H NMR (500 MHz, CDCl 3)δ10.86(s,1H),8.65(dd,J=8.5,4.4Hz,1H),8.04(s,1H),7.47(t,J=7.8Hz,1H),7.28-7.21(m,1H),7.11-7.05(m ,2H),6.91(dd,J=8.6,1.9Hz,1H),6.81(s,1H),6.71(d,J=8.7Hz,1H),4.18(dd,J=10.5,2.0Hz,1H),4.01(t,J=9.7 Hz,1H),3.65(d,J=11.2Hz,1H),3.12(t,J=9.5Hz,1H),3.01(d,J=10.8Hz,1H),2.95(d,J=11.3Hz,2H),2.86(d,J=1 0.9Hz,1H),2.79-2.66(m,1H),2.35-2.30(m,3H),2.06-1.95(m,2H),1.83(s,3H),1.81(s,3H),1.69-1.50(m,2H). MS m / z582.5[M+H] + .

[0233] Example 16: Preparation of Compound 12R

[0234]

[0235] KOH (286 mg, 5.10 mmol) was added to a dry DMSO (6 mL) solution of 1,2-difluoro-4-nitrobenzene (1a, 326 mg, 2.05 mmol) and (S)-3-hydroxymethylmorpholine (12R-1a, 200 mg, 1.70 mmol). The mixture was stirred at room temperature for 3 hours and then heated to 60 ° C for 5 hours. TLC monitoring showed that the reaction raw materials were almost consumed. The reaction mixture was cooled to room temperature, ice water was added, and extracted with dichloromethane (20 mL x 3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6: 1) to obtain a yellow solid compound 12R-1b (187 mg, yield 46%).

[0236] Palladium-carbon catalyst (10%, 50 mg) was added to a methanol (10 mL) solution of compound 12R-1b at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and under a hydrogen (1 atmosphere) atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 50: 1) to obtain a colorless oily compound 12R-1c (38 mg, yield 24%). MS m / z 207.8 [M+H] + .

[0237] To a solution of compound 12R-1c (38 mg, 0.18 mmol) and compound 1k (58 mg, 0.18 mmol) in 2-methoxyethanol (1.5 mL) was added HCl methanol solution (2.5 M, 0.22 mL). The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain a yellow solid compound 12R (30 mg, yield 34%). 1 H NMR (500 MHz, CDCl 3 )δ10.87(s,1H),8.65(dd,J=8.5,4.4Hz,1H),8.06(s,1H),7.48(t,J=8.6,1H),7.33-7.20(m,3H),7.13-7.06(m, 2H),6.93(dd,J=8.6,2.5Hz,1H),6.77(s,1H),6.69(d,J=8.7Hz,1H),4.16(dd,J=10.5,2.7Hz,1H),4.05(dd,J=1 1.5,3.5Hz,1H),3.98(dd,J=10.5,9.1Hz,1H),3.88(dd,J=10.9,3.1Hz,1H),3.76(td,J=11.6,2.8Hz,1H),3.51- 3.45(m,1H),3.30(t,J=10.7Hz,1H),3.24-3.18(m,1H),2.84(td,J=11.8,3.6Hz,1H),1.84(s,3H),1.81(s,3H). MS m / z 486.9[M+H] + .

[0238] Example 17: Preparation of Compound 12S

[0239]

[0240] KOH (286 mg, 5.10 mmol) was added to a solution of 1,2-difluoro-4-nitrobenzene (1a, 317 mg, 2.05 mmol) and (R)-3-hydroxymethylmorpholine (12S-1a, 200 mg, 1.70 mmol) in dry DMSO (6 mL). The mixture was stirred at room temperature for 3 hours and then heated to 60 ° C for 5 hours. TLC monitoring showed that the reaction raw materials were almost consumed. The reaction mixture was cooled to room temperature, ice water was added, and extracted with dichloromethane (25 mL x 3). The combined organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to obtain a yellow solid compound 12S-1b. The compound was directly used in the next step.

[0241] Palladium-carbon catalyst (10%, 100 mg) was added to a methanol (15 mL) solution of compound 12S-1b at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight at room temperature and under a hydrogen (1 atmosphere) atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 50: 1) to obtain a colorless oily compound 12S-1c (44 mg, two-step total yield: 13%). MSm / z 207.4 [M+H] + .

[0242] To a solution of compound 12S-1c (44 mg, 0.21 mmol) and compound 1k (68 mg, 0.21 mmol) in 2-methoxyethanol (2 mL) was added HCl methanol solution (2.5 M, 0.22 mL). The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain a yellow solid compound 12S (11 mg, yield 11%). 1 H NMR (400 MHz, CDCl 3)δ10.87(s,1H),8.65(dd,J=8.4,4.4Hz,1H),8.05(s,1H),7.48(t,J=7.9Hz,1H),7.30-7.21(m,1H),7.14-7.0 6(m,2H),6.93(dd,J=8.7,2.5Hz,1H),6.83(s,1H),6.69(d,J=8.7Hz,1H),4.16(dd,J=10.5,2.6Hz,1H),4.05( dd,J=11.5,3.2Hz,1H),3.98(dd,J=10.3,9.2Hz,1H),3.88(dd,J=10.8,2.8Hz,1H),3.76(td,J=11.6,2.8Hz,1 H),3.53-3.44(m,1H),3.30(t,J=10.6Hz,1H),3.25-3.15(m,1H),2.89-2.77(m,1H),1.84(s,3H),1.81(s,3H). MS m / z486.5[M+H] + .

[0243] Example 18: Preparation of Compound 13R

[0244]

[0245] Trifluoroacetic acid (4.5 mL) was slowly added to a solution of compound 4R-1a (1.5 g, 4.5 mmol) in dichloromethane (10 mL) at 0°C. The reaction was stirred at room temperature for 1 h. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure and neutralized by adding saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (20 mL x 3). The combined organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product 13R-1a (3.06 g). The crude product was used directly in the next step.

[0246] DIPEA (3.36 g, 26.0 mmol) was added to a solution of crude product 13R-1a (3.06 g) in dichloromethane (20 mL) at 0°C with stirring. Trifluoroacetic anhydride (3.0 g, 14.3 mmol) was then slowly added dropwise thereto. The reaction was then moved to room temperature and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and then a saturated sodium bicarbonate solution was added to adjust the pH to about 8, and then extracted with dichloromethane (30 mL x 3). The combined organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give a yellow solid compound 13R-1b (1.2 g, two-step total yield 63%). MS m / z 332.5[M+H] + .

[0247] To a solution of compound 13R-1b (950 mg, 2.87 mmol) in THF (10 mL) was added BH 3 -THF (1.0M, 7.17mL). The reaction mixture was stirred at reflux for 2h. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain yellow solid compound 13R-1c. The crude product was directly used for the next step reaction.

[0248] Palladium-carbon catalyst (10%, 200 mg) was added to a methanol (20 mL) solution of compound 13R-1c at room temperature. The reaction system was replaced with hydrogen three times. The reaction mixture was stirred overnight under a hydrogen (1 atmosphere) atmosphere at room temperature. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 50: 1 to 20: 1) to obtain a yellow solid compound 13R-1d (520 mg, two-step total yield 63%). MS m / z 288.4 [M+H] + .

[0249] To a solution of compound 13R-1d (91 mg, 0.32 mmol) and compound 1k (100 mg, 0.32 mmol) in 2-methoxyethanol (2 mL) was added HCl methanol solution (2.5 M, 0.33 mL). The reaction mixture was placed in a sealed tube, heated to 120 ° C, and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain a yellow solid compound 13R (71 mg, yield 40%). 1 H NMR (500 MHz, CDCl 3)δ10.80(s,1H),8.58(dd,J=8.3,4.4Hz,1H),7.99(s,1H),7.41(dd,J=8.4,7.4Hz,1H),7.22-7.15(m,1H),7.05 -7.00(m,2H),6.85(dd,J=8.7,2.5Hz,1H),6.69(s,1H),6.65(d,J=8.8Hz,1H),4.11(dd,J=10.5,2.7Hz,1H),3.9 4(dd,J=10.5,8.9Hz,1H),3.56(dt,J=11.4,2.4Hz,1H),3.16-3.08(m,1H),3.02-2.93(m,3H),2.88-2.82(m,1H ), 2.77(td,J=11.6,3.1Hz,1H),2.62(td,J=11.3,2.9Hz,1H),2.21(t,J=10.6Hz,1H),1.77(s,3H),1.74(s,3H). MS m / z 567.6[M+H] + .

[0250] Example 19: Preparation of Compound 14R

[0251]

[0252] Compound 4R (50 mg, 0.10 mmol), dimethylaminoethyl chloride hydrochloride (14R-1a, 16 mg, 0.10 mmol), sodium iodide (16 mg, 0.10 mmol) and potassium carbonate (42 mg, 0.30 mmol) were added to the sealed tube in sequence, and finally N, N-dimethylformamide (2 mL) was added to dissolve it. The reaction mixture was heated to 80 ° C in a sealed tube and stirred overnight. TLC monitored the reaction and there was still raw material remaining. The reaction mixture was cooled to room temperature, water (10 mL) was added, and it was extracted with dichloromethane (15 mL x 3). The combined organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 3: 1) to obtain an off-white solid compound 14R (2 mg, yield 4%). 1 H NMR (500 MHz, CDCl 3)δ10.89(s,1H),8.65(dd,J=8.3,4.5Hz,1H),8.05(s,1H),7.48(t,J=7.5Hz,1H),7.26(d,J=20.4Hz,1H),7.15-7.07( m,2H),6.92(dd,J=8.6,2.0Hz,1H),6.79(s,1H),6.71(d,J=8.8Hz,1H),4.20(dd,J=10.4,2.2Hz,1H),4.05-3.97(m,1H ),3.63(d,J=11.6Hz,2H),3.19(dd,J=10.3,9.0Hz,1H),3.04(d,J=10.3Hz,1H),2.91(d,J=10.1Hz,1H),2.82(td,J=11 .5,2.5Hz,1H),2.73-2.63(m,2H),2.56-2.39(s,6H),2.37-2.29(m,2H),2.24-2.18(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 556.5[M+H] + .

[0253] Example 20: Preparation of Compound 15R

[0254]

[0255] Compound 4R-2 (48 mg, 0.10 mmol), palladium acetate (0.2 mg, 0.001 mmol), RuPhos (0.9 mg, 0.002 mmol), sodium tert-butoxide (11.5 mg, 0.12 mmol) and 3-bromopyridine (15R-1a, 17.4 mg, 0.11 mmol) were added to the sealed tube in sequence, and N,N-dimethylformamide (0.5 mL) was added at last. The reaction mixture was heated to 110 ° C in a sealed tube and stirred overnight. TLC monitored the reaction and there was still raw material remaining. The reaction mixture was cooled to room temperature, water (10 mL) was added, and it was extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by preparative thin-layer chromatography (dichloromethane:methanol=20:1, 2% triethylamine) to obtain yellow solid compound 15R (6 mg, yield 11%). MS m / z 562.5[M+H] + .

[0256] Example 21: Preparation of Compound 16R

[0257]

[0258] Compound 4R-2 (48 mg, 0.10 mmol), palladium acetate (0.2 mg, 0.001 mmol), RuPhos (0.9 mg, 0.002 mmol), sodium tert-butoxide (11.5 mg, 0.12 mmol) and bromobenzene (16R-1a, 17.3 mg, 0.11 mmol) were added to the sealed tube in sequence, and N,N-dimethylformamide (0.5 mL) was added at last. The reaction mixture was heated to 110 °C in a sealed tube and stirred overnight. TLC monitored the reaction and there was still raw material remaining. After the reaction mixture was cooled to room temperature, water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was separated and purified by preparative thin layer chromatography (dichloromethane:methanol=20:1, 2% triethylamine) to obtain yellow solid compound 16R (4 mg, yield 7%). 1 H NMR (400 MHz, CDCl 3 )δ10.82(s,1H),8.59(dd,J=8.5,4.4Hz,1H),7.99(s,1H),7.43(dd,J=8.7,7.7Hz,1H),7.27-7. 16(m,3H),7.07(d,J=2.5Hz,1H),7.03(td,J=7.4,1.4Hz,1H),6.95-6.82(m,4H),6.75-6.69(m, 2H),4.24(dd,J=10.5,2.6Hz,1H),4.05(dd,J=10.5,9.0Hz,1H),3.75-3.62(m,2H),3.55-3.48( m,1H),3.28-3.20(m,1H),2.97-2.82(m,2H),2.53(t,J=11.1Hz,1H),1.78(s,3H),1.75(s,3H). MS m / z561.6[M+H] + .

[0259] Example 22: Preparation of Compound 17R

[0260]

[0261] At room temperature, compound 4R (48.5 mg, 0.1 mmol), 17R-1a (10.5 mg, 0.15 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (57 mg, 0.15 mmol) and N,N-diisopropylethylamine (20 mg, 0.15 mmol) were dissolved in DMF (1.5 mL), and the reaction mixture was stirred for 0.5 h. TLC monitored the completion of the reaction. Water (10 mL) was added to the reaction mixture, and it was extracted with dichloromethane (15 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 10: 1) to obtain yellow solid compound 17R. MS m / z 537.6 [M+H] + .

[0262] Example 23: Preparation of Compound 18R

[0263]

[0264] Compound 13R-1a (400 mg, 1.70 mmol), 18R-1a (422 mg, 2.55 mmol), O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethylurea (970 mg, 2.55 mmol) and diisopropylethylamine (330 mg, 2.55 mmol) were dissolved in acetonitrile (20 mL), and the reaction mixture was stirred at room temperature for 30 minutes. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain a yellow solid compound 18R-1b (417 mg, yield 71%). MS m / z 347.5 [M+H] + .

[0265] Compound 18R-1b (326 mg, 0.94 mmol) and tin dichloride dihydrate (977 mg, 4.71 mmol) were dissolved in ethanol (8 mL), and then HCl aqueous solution (2.0 M, 2.35 mL) was added, and the reaction mixture was stirred at 90 ° C overnight. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, redissolved in a small amount of dichloromethane, saturated sodium bicarbonate solution was added to adjust the pH to 7-8, and extracted with dichloromethane (20x 3 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid compound 18R-1c (203 mg, yield 68%). MS m / z 317.4 [M+H] + .

[0266] Compound 18R-1c (80 mg, 0.25 mmol) and compound 1k (80 mg, 0.25 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.26 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 9: 1) to obtain a light yellow solid compound 18R (14 mg, yield 9%). 1 H NMR (500 MHz, CDCl 3 )δ10.90(s,1H),8.66-8.62(m,1H),8.06(s,1H),7.49(t,J=7.9Hz,1H),7.30-7.24(m,1H),7.1 5(d,J=2.4Hz,1H),7.14-7.10(m,1H),6.96-6.92(m,1H),6.91-6.81(m,2H),6.80(s,1H),6.76- 6.70(m,1H),4.79-4.60(m,1H),4.39-4.13(m,2H),4.08-4.02(m,1H),3.76(d,J=11.6Hz,1H),3 .51-3.34(m,3H),3.15-2.91(m,2H),2.77-2.70(m,1H),2.56(s,6H),1.84(s,3H),1.82(s,3H). MS m / z596.5[M+H] + .

[0267] Example 24: Preparation of Compound 19R

[0268]

[0269] Compound 13R-1a (500 mg, 2.13 mmol) and triethylamine (258 mg, 2.55 mmol) were dissolved in dichloromethane (15 mL) at 0°C, and then compound 19R-1a (416 mg, 2.55 mmol) in dichloromethane (5 mL) was slowly added, and the reaction mixture was stirred at room temperature for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1 to 1:2) to obtain yellow solid compound 19R-1b (153 mg, yield 22%). MS m / z 326.3 [M+H] + .

[0270] Compound 19R-1b (150 mg, 0.46 mmol) and tin dichloride dihydrate (479 mg, 2.31 mmol) were dissolved in ethanol (8 mL), and then HCl aqueous solution (2.0 M, 1.15 mL) was added, and the reaction mixture was stirred at 90 ° C overnight. TLC monitoring of the reaction was complete. The reaction mixture was concentrated under reduced pressure, dissolved in a small amount of dichloromethane, saturated sodium bicarbonate solution was adjusted to pH 7-8, and extracted with dichloromethane (20 x 3 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was then separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a yellow solid compound 19R-1c (107 mg, yield 78%). 1 H NMR (500 MHz, DMSO-d 6 )δ6.84(dd,J=16.5,10.0Hz,1H),6.60(d,J=8.6Hz,1H),6.21(d,J=10.0Hz,1H),6.15(d,J =16.5Hz,1H),6.09(dd,J=8.5,2.4Hz,1H),6.03(d,J=2.4Hz,1H),4.59(bs,2H),4.22(dd, J=10.6,2.5Hz,1H),3.82(dd,J=10.5,8.7Hz,1H),3.72(d,J=12.1Hz,1H),3.51(t,J=10.2 Hz, 2H), 2.96-2.90 (m, 1H), 2.81-2.74 (m, 1H), 2.57-2.51 (m, 1H), 2.42 (t, J = 11.1Hz, 1H).

[0271] Compound 19R-1c (50 mg, 0.17 mmol) and compound 1k (54 mg, 0.17 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.18 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 50: 1, 2% triethylamine) to obtain a light yellow solid compound 19R (32 mg, yield 32%). 1 H NMR (500 MHz, CDCl 3)δ10.93(s,1H),8.65-8.61(m,1H),8.05(s,1H),7.50-7.45(m,1H),7.30-7.24(m,1H),7.16(d,J=2.5Hz,1 H),7.14-7.09(m,1H),7.03(s,1H),6.96-6.92(m,1H),6.73(d,J=8.8Hz,1H),6.44(dd,J=16.6,9.9Hz,1H) ,6.30(d,J=16.6Hz,1H),6.10(d,J=9.9Hz,1H),4.26-4.22(m,1H),4.04-3.99(m,1H),3.81-3.74(m,2H),3 .68-3.64(m,1H),3.31-3.23(m,1H),2.94-2.84(m,2H),2.54(t,J=11.1Hz,1H),1.84(s,3H),1.82(s,3H). MS m / z 575.5[M+H] + .

[0272] Example 25: Preparation of Compound 20R

[0273]

[0274] 4-Bromocrotonic acid (20R-1a, 1.0 g, 6.06 mmol) was dissolved in methanol (60 mL) at room temperature, and a methanol solution of sodium methoxide (1.64 g, 30.3 mmol, 10 mL) was added. The reaction mixture was stirred at room temperature for 30 minutes and then stirred under reflux for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure, then dissolved in a mixture of water and ethyl acetate (v / v = 50 / 50), and HCl aqueous solution (2.0 M) was added dropwise to adjust the pH to 1, and extracted with ethyl acetate (3 x 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 2: 1) to obtain a yellow solid compound 20R-1b (622 mg, yield 88%). 1 H NMR (500 MHz, CDCl 3 )δ7.06(dt,J=15.7,4.1Hz,1H),6.08(dt,J=15.7,2.1Hz,1H),4.12(dd,J=4.1,2.1Hz,2H),3.41(s,3H).

[0275] Compound 13R-1a (200 mg, 0.85 mmol), 4-methoxycrotonic acid (20R-1b, 148 mg, 1.28 mmol), O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethylurea (487 mg, 1.28 mmol) and diisopropylethylamine (165 mg, 1.28 mmol) were dissolved in acetonitrile (15 mL), and the reaction mixture was stirred at room temperature for 30 minutes. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain a yellow solid compound 20R-1c (700 mg) which was directly used in the next step. MS m / z 334.3 [M+H] + .

[0276] Compound 20R-1c (700 mg, 2.10 mmol) and iron powder (586 mg, 10.5 mmol) were added to a mixture of ethyl acetate / water (v / v = 10 mL / 6 mL), and ammonium chloride (702 mg, 13.12 mmol) was added, and the reaction mixture was stirred at room temperature for 4 hours. TLC monitored the completion of the reaction. The reaction mixture was filtered, and the filtrate was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was then separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain a yellow solid compound 20R-1d (55 mg, two-step yield 21%). MS m / z 304.4 [M+H] + .

[0277] Compound 20R-1d (55 mg, 0.18 mmol) and compound 1k (60 mg, 0.18 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.20 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to obtain yellow solid compound 20R (8.3 mg, yield 8%). 1 H NMR (500 MHz, CDCl 3)δ10.95(s,1H),8.66-8.63(m,1H),8.04(s,1H),7.48(t,J=7.9Hz,1H),7.30-7.24(m,1H) ,7.20-7.09(m,2H),7.04(s,1H),6.98-6.89(m,2H),6.77-6.70(m,1H),6.61-6.51(m,1H) ,4.82-4.60(m,1H),4.32-4.25(m,1H),4.17-3.90(m,4H),3.73(d,J=11.0Hz,1H),3.43(s ,3H),3.11-2.92(m,2H),2.77-2.66(m,1H),2.61-2.50(m,1H),1.84(s,3H),1.82(s,3H). MS m / z 583.6[M+H] + .

[0278] Example 26: Preparation of Compound 21R

[0279]

[0280] Compound 13R-1a (300 mg, 1.28 mmol) was dissolved in dichloromethane (10 mL), and N-Boc-piperidone (21R-1a, 381 mg, 1.91 mmol) and 2 drops of acetic acid were added in sequence. The reaction mixture was stirred at room temperature for 2 hours. Then sodium triacetoxyborohydride (405 mg, 1.91 mmol) was slowly added, and then stirred at room temperature overnight. After the reaction was completed, 1N sodium hydroxide solution was added to the reaction mixture at 0°C, and the mixture was poured into ice water and extracted with dichloromethane (3x 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain yellow solid compound 21R-1b (483 mg, yield 91%). MS m / z 419.5[M+H] + .

[0281] Compound 21R-1b (200 mg, 0.48 mmol) and palladium carbon catalyst (10%, 50 mg) were added to methanol (20 mL) at room temperature. The reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain a colorless oily compound 21R-1c (77 mg, yield 41%). MS m / z 389.6 [M+H] + .

[0282] Compound 21R-1c (77 mg, 0.20 mmol) and compound 1k (63 mg, 0.20 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.21 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 3:1, 2% triethylamine) to obtain yellow solid compound 21R (26 mg, yield 23%). 1 HNMR (500MHz, CDCl 3 )δ10.89(s,1H),8.67-8.63(m,1H),8.04(s,1H),7.51-7.45(m,1H),7.29-7.22(m,1H),7.12-7.07( m,2H),6.92(dd,J=8.8,2.5Hz,1H),6.88(s,1H),6.71(d,J=8.8Hz,1H),4.22-4.14(m,1H),4.06-3. 93(m,1H),3.66(d,J=11.4Hz,1H),3.50-3.39(m,2H),3.14-3.06(m,1H),2.98(d,J=10.5Hz,1H),2. 92-2.81(m,3H),2.78-2.70(m,1H),2.60-2.47(m,3H),2.11-1.88(m,4H),1.84(s,3H),1.81(s,3H). MS m / z 568.5[M+H] + .

[0283] Example 27: Preparation of Compound 22R

[0284]

[0285] Compound 21R-1b (283 mg, 0.73 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (1.5 mL) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 10:1, 2% triethylamine) to obtain yellow solid compound 22R-1a (150 mg, yield 65%). MS m / z 319.4 [M+H] + .

[0286] Compound 22R-1a (150 mg, 0.47 mmol) and diisopropylethylamine (67 mg, 0.52 mmol) were dissolved in dichloromethane (8 mL), and a dichloromethane dilution (2 mL) of acryloyl chloride (47 mg, 0.52 mmol) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 15:1) to obtain a yellow solid compound 22R-1b (240 mg) which was directly used in the next step. MS m / z 373.5 [M+H] + .

[0287] Compound 22R-1b (240 mg, 0.64 mmol) and iron powder (180 mg, 3.22 mmol) were dissolved in a mixed solvent of ethyl acetate / water (v / v=5 mL / 3 mL), and ammonium chloride (215 mg, 4.03 mmol) was added. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was filtered through diatomaceous earth, and the filtrate was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain a yellow solid compound 22R-1c (91 mg, two-step yield 56%). MS m / z 343.5 [M+H] + .

[0288] Compound 22R-1c (91 mg, 0.27 mmol) and compound 1k (85 mg, 0.27 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.28 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 10: 1) to obtain yellow solid compound 22R (12 mg, yield 7%). 1 H NMR (500 MHz, CDCl 3)δ10.91(s,1H),8.67-8.63(m,1H),8.04(s,1H),7.48(t,J=7.9Hz,1H),7.29-7.22(m,1H),7.12-7.06(m,2H),6.93(dd,J=8 .8,2.5Hz,1H),6.89(s,1H),6.72(d,J=8.8Hz,1H),6.58(dd,J=16.8,10.6Hz,1H),6.28(dd,J=16.8,1.8Hz,1H),5.69(dd,J =10.6,1.8Hz,1H),4.72(d,J=10.5Hz,1H),4.24-4.16(m,1H),4.11-3.98(m,2H),3.68(d,J=11.4Hz,1H),3.27-2.96(m,3H) ,2.93-2.75(m,2H),2.73-2.48(m,3H),2.19-2.08(m,1H),2.00-1.88(m,2H),1.74-1.57(m,2H),1.84(s,3H),1.81(s,3H). MS m / z 622.6[M+H] + .

[0289] Example 28: Preparation of Compound 23R

[0290]

[0291] Compound 23R-1a (5.0 g, 21.01 mmol), 23R-1b (4.54 g, 21.01 mmol) and potassium hydroxide (3.54 g, 63.03 mmol) were dissolved in dimethyl sulfoxide (60 mL), stirred at room temperature for 3 hours, and then stirred at 60 ° C for 3 hours. After the reaction was completed, the reaction mixture was cooled and poured into ice water, and stirred at room temperature for 1 hour. The mixture was then extracted with dichloromethane (3x 40 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain yellow solid compound 23R-1c (5.0 g, yield 57%) and yellow solid compound 24R-1a (1.4 g, yield 16%). 23R-1c: 1 H NMR (500 MHz, CDCl 3)δ8.04-8.02(m,1H),7.72-7.70(m,1H),4.25-4.14(m,2H),3.87-3.76(m,2H), 3.64-3.55(m,1H),3.51-3.32(m,2H),3.29-3.16(m,2H),1.49(s,9H); 24R-1a: 1 H NMR (500 MHz, CDCl 3 )δ7.94(d,J=2.4Hz,1H),7.63(d,J=2.2Hz,1H),4.50-4.43(m,1H),4.28-4.05(m,3H),3.75-3.70(m,1H ),3.22–3.15(m,1H),3.12-2.99(m,1H),2.84(td,J=11.8,2.9Hz,1H),2.69-2.56(m,1H),1.49(s,9H).

[0292] A mixture of compound 23R-1c (2.0 g, 4.83 mmol), tetramethyltin (1.73 g, 9.66 mmol), tetrakistriphenylphosphine palladium (279 mg, 0.24 mmol) and lithium chloride (409 mg, 9.66 mmol) in N,N-dimethylformamide (35 mL) was heated to 95°C and stirred overnight. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain yellow solid compound 23R-1d (1.37 g, yield 81%). 1 H NMR (500 MHz, CDCl 3 )δ7.64(d,J=2.5Hz,1H),7.58(d,J=2.6Hz,1H),4.25-4.13(m,2H),3.87-3.80(m,2H),3. 60-3.52(m,1H),3.35(s,1H),3.23(s,1H),3.10-2.97(m,2H),2.36(s,3H),1.48(s,9H).

[0293] Compound 23R-1d (200 mg, 0.57 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (8 mL) at room temperature, and the reaction mixture was stirred for 2 hours under a hydrogen atmosphere at room temperature and 1 atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain an off-white solid compound 23R-1e (162 mg, 89%). 1 H NMR (500 MHz, CDCl3 )δ6.14(d,J=2.4Hz,1H),6.10(d,J=2.4Hz,1H),4.31(t,J=10.9Hz,1H),4.17-3.93(m,3H),3.34 (s,1H),3.20-2.97(m,2H),2.90(d,J=12.2Hz,1H),2.79-2.66(m,1H),2.20(s,3H),1.47(s,9H).

[0294] Compound 23R-1e (50 mg, 0.16 mmol) and compound 1k (50 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.16 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 10: 1, 2% triethylamine) to obtain a light yellow solid compound 23R (59 mg, yield 75%). 1 H NMR (500 MHz, CDCl 3 )δ10.86(s,1H),8.67-8.63(m,1H),8.07(s,1H),7.53-7.47(m,1H),7.30-7.23(m,1H) ,7.13-7.09(m,1H),7.08(d,J=2.4Hz,1H),6.92(s,1H),6.81(d,J=2.3Hz,1H),4.67(t ,J=10.8Hz,1H),4.05(dd,J=10.8,3.0Hz,1H),3.35(dd,J=12.8,4.3Hz,1H),3.23-3.1 8(m,1H),3.17-3.00(m,4H),2.94-2.87(m,1H),2.25(s,3H),1.84(s,3H),1.82(s,3H). MS m / z 499.4[M+H] + .

[0295] Example 29: Preparation of Compound 23S

[0296]

[0297] Compound 23R-1a (100 mg, 0.42 mmol), 23S-1a (91 mg, 0.42 mmol) and potassium hydroxide (71 mg, 1.26 mmol) were dissolved in dimethyl sulfoxide (5 mL), stirred at room temperature for 3 hours, and then stirred at 60 ° C for 3 hours. After the reaction was completed, the reaction mixture was cooled and poured into ice water, and stirred at room temperature for 1 hour. The mixture was then extracted with dichloromethane (3x25 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain yellow solid compound 23S-1b (64 mg, yield 37%) and yellow solid compound 24S-1a (64 mg, yield 37%). 23S-1b: 1 H NMR (500 MHz, CDCl 3 )δ8.03(d,J=2.5Hz,1H),7.71(d,J=2.6Hz,1H),4.26-4.15(m,2H),3.86-3.78(m,2H),3 .60(dd,J=13.8,4.1Hz,1H),3.52-3.36(m,2H),3.29-3.15(m,2H),1.48(s,9H); 24S-1a: 1 H NMR (500 MHz, CDCl 3 )δ7.94(d,J=2.5Hz,1H),7.63(d,J=2.5Hz,1H),4.49-4.44(m,1H),4.28-4.04(m,3H),3.71(d,J=11 .3Hz,1H),3.22-3.15(m,1H),3.10-3.01(m,1H),2.88-2.79(m,1H),2.66-2.57(m,1H),1.49(s,9H).

[0298] A mixture of compound 23S-1b (64 mg, 0.16 mmol), tetramethyltin (56 mg, 0.31 mmol), tetrakistriphenylphosphine palladium (9 mg, 0.01 mmol) and lithium chloride (13 mg, 0.31 mmol) in N,N-dimethylformamide (2 mL) was heated to 90°C and stirred overnight. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain yellow solid compound 23S-1c (33 mg, yield 60%). 1 H NMR (500 MHz, CDCl 3)δ7.65(d,J=2.5Hz,1H),7.59(d,J=2.6Hz,1H),4.24-4.14(m,2H),3.89-3.78(m,2H),3.60-3. 49(m,1H),3.39-3.30(m,1H),3.28-3.18(m,1H),3.10-2.98(m,2H),2.37(s,3H),1.48(s,9H).

[0299] Compound 23S-1c (33 mg, 0.09 mmol) and palladium carbon catalyst (10%, 40 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction, the reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was then separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain a light yellow solid compound 23S-1d (23 mg, 76%). MS m / z 320.4 [M+H] + .

[0300] Compound 23S-1d (23 mg, 0.07 mmol) and compound 1k (23 mg, 0.07 mmol) were dissolved in 2-methoxyethanol (1 mL), and HCl methanol solution (2.5 M, 0.08 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 5:1) to obtain a light yellow solid compound 23S (14 mg, yield 38%). 1 H NMR (500MHz, CD 3 OD)δ8.50-8.45(m,1H),8.04(s,1H),7.62-7.53(m,2H),7.27-7.22(m,1 H),6.98(d,J=2.4Hz,1H),6.82(d,J=2.1Hz,1H),4.53(t,J=11.0Hz,1H), 4.02(dd,J=10.7,2.8Hz,1H),3.30-3.26(m,1H),3.18-3.09(m,2H),3.07 -2.99(m,3H),2.87-2.80(m,1H),2.20(s,3H),1.86(s,3H),1.83(s,3H). MS m / z499.4[M+H] + .

[0301] Example 30: Preparation of Compound 24R

[0302]

[0303] A mixture of compound 24R-1a (1.0 g, 2.41 mmol), tetramethyltin (863 mg, 4.83 mmol), tetrakistriphenylphosphine palladium (140 mg, 0.12 mmol) and lithium chloride (205 mg, 4.83 mmol) in N,N-dimethylformamide (15 mL) was heated to 95°C and stirred overnight. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain yellow solid compound 24R-1b (861 mg, yield 100%). 1 H NMR (500 MHz, CDCl 3 )δ7.60-7.54(m,2H),4.38(dd,J=10.8,2.8Hz,1H),4.26-4.09(m,2H),4.06(dd,J=10.8,8.8Hz,1H),3 .73-3.68(m,1H),3.14-3.01(m,2H),2.82-2.73(m,1H),2.65-2.55(m,1H),2.21(s,3H),1.49(s,9H).

[0304] Compound 24R-1b (150 mg, 0.43 mmol) and palladium carbon catalyst (10%, 50 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 2 hours under a hydrogen atmosphere at room temperature and 1 atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain an off-white solid compound 24R-1c (119 mg, 87%). 1 H NMR (500 MHz, CDCl 3 )δ6.11(d,J=2.3Hz,1H),6.05(d,J=2.0Hz,1H),4.19(dd,J=10.7,2.7Hz,1H),4.15-3.98(m,2H),3.91(dd,J=10.6,8.4Hz,1H ),3.62-3.57(m,1H),3.10-3.03(m,1H),3.02-2.97(m,1H),2.74-2.67(m,1H),2.64-2.59(m,1H),2.09(s,3H),1.48(s,9H).

[0305] Compound 24R-1c (50 mg, 0.16 mmol) and compound 1k (50 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.16 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 10: 1, 2% triethylamine) to obtain a light yellow solid compound 24R (52 mg, yield 66%). 1 H NMR (500 MHz, DMSO-d 6 )δ11.14(s,1H),9.04(s,1H),8.67-8.52(m,1H),8.14(s,1H),7.61-7.55(m,1H),7.46 (t,J=7.8Hz,1H),7.16(t,J=7.4Hz,1H),6.98(s,1H),6.91(s,1H),4.32-4.28(m,1H), 3.96-3.91(m,1H),3.59(d,J=12.3Hz,1H),3.30-3.24(m,3H),2.92(t,J=12.1Hz,1H), 2.83(t,J=12.4Hz,1H),2.69(t,J=11.8Hz,1H),2.05(s,3H),1.80(s,3H),1.77(s,3H). MS m / z 499.4[M+H] + .

[0306] Example 31: Preparation of Compound 24S

[0307]

[0308] A mixture of compound 24S-1a (39 mg, 2.41 mmol), tetramethyltin (34 mg, 0.19 mmol), tetrakistriphenylphosphine palladium (6 mg, 0.004 mmol) and lithium chloride (8 mg, 0.19 mmol) in N,N-dimethylformamide (2 mL) was heated to 95°C and stirred overnight. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain yellow solid compound 24S-1b (28 mg, yield 85%). 1 H NMR (500 MHz, CDCl 3)δ7.60-7.55(m,2H),4.40-4.35(m,1H),4.25-4.02(m,3H),3.71(d,J=11.4Hz,1H), 3.16-3.00(m,2H),2.83-2.73(m,1H),2.66-2.57(m,1H),2.21(s,3H),1.49(s,9H).

[0309] Compound 24S-1b (28 mg, 0.08 mmol) and palladium carbon catalyst (10%, 40 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain an off-white solid compound 24S-1c (12 mg, 47%). MS m / z 320.4 [M+H] + .

[0310] Compound 24S-1c (12 mg, 0.04 mmol) and compound 1k (12 mg, 0.04 mmol) were dissolved in 2-methoxyethanol (1 mL), and HCl methanol solution (2.5 M, 0.04 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 5:1) to obtain yellow solid compound 24S (8 mg, yield 42%). 1 H NMR (500MHz, CD 3 OD)δ8.52-8.48(m,1H),8.02(s,1H),7.62-7.55(m,1H),7.51(t,J=7.9Hz,1H),7.25-7 .20(m,1H),6.89(d,J=2.0Hz,1H),6.74(d,J=1.9Hz,1H),4.27-4.21(m,1H),3.96-3.9 0(m,1H),3.50(d,J=11.9Hz,1H),3.08-3.01(m,3H),2.89-2.80(m,1H),2.63-2.56(m, 1H), 2.51 (t, J = 11.7Hz, 1H), 2.07 (s, 3H), 1.87 (d, J = 2.9Hz, 3H), 1.84 (d, J = 2.9Hz, 3H). MS m / z 499.4[M+H] + .

[0311] Example 32: Preparation of Compound 25R

[0312]

[0313] Compound 23R-1d (1.17 g, 3.35 mmol) was dissolved in methanol (15 mL) at room temperature, and HCl methanol solution (4.0 M, 1.36 mL) was added dropwise, and the mixture was stirred at 60 ° C for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, dissolved in dichloromethane, and neutralized with saturated sodium bicarbonate aqueous solution. The mixture was extracted with dichloromethane (3x 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain yellow solid compound 25R-1a (465 mg, yield 56%). 1 H NMR (500 MHz, CDCl 3 )δ7.57(d,J=2.5Hz,1H),7.51(d,J=2.6Hz,1H),4.37-4.27(m,1H),4.10-4.01(m,1H),3.19-3.12(m,2H),3.09-2.91(m,5H),2.29(s,3H).

[0314] A mixture of compound 25R-1a (150 mg, 0.60 mmol), formaldehyde solution (37%, 1.5 mL) and 2 drops of acetic acid in methanol (6 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (113 mg, 1.81 mmol) was then added, and the reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and water (15 mL) was added and then extracted with dichloromethane (3x15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain yellow solid compound 25R-1b (155 mg, yield 98%). MS m / z 264.4 [M+H] + .

[0315] Compound 25R-1b (155 mg, 0.59 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (8 mL) at room temperature, and the reaction mixture was stirred for 2 hours under a hydrogen atmosphere at room temperature and 1 atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain a light yellow solid compound 25R-1c (123 mg, 90%). 1 H NMR (500 MHz, CDCl 3)δ6.12(d,J=2.6Hz,1H),6.09(d,J=2.6Hz,1H),4.62(t,J=10.7Hz,1H),4.01-3.96(m,1H),3.42(bs,2H),3. 18(d,J=10.6Hz,1H),3.02-2.76(m,4H),2.59(d,J=9.4Hz,1H),2.37-2.33(m,1H),2.31(s,3H),2.19(s,3H).

[0316] Compound 25R-1c (50 mg, 0.22 mmol) and compound 1k (68 mg, 0.22 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.22 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to obtain a light yellow solid compound 25R (73 mg, yield 66%). 1 HNMR (500MHz, CDCl 3 )δ10.89(s,1H),8.68-8.64(m,1H),8.07(s,1H),7.49(t,J=7.9Hz,1H),7.30-7.22(m,1H),7.12-7. 05(m,2H),6.96(s,1H),6.80(d,J=2.3Hz,1H),4.62(t,J=10.7Hz,1H),4.05(dd,J=10.8,2.8Hz,1H), 3.21(d,J=10.8Hz,1H),3.07-3.02(m,1H),2.98-2.91(m,1H),2.87(d,J=11.8Hz,1H),2.80(d,J=10. 0Hz,1H),2.63-2.56(m,1H),2.39-2.32(m,1H),2.31(s,3H),2.25(s,3H),1.84(s,3H),1.81(s,3H). MS m / z 513.4[M+H] + .

[0317] Example 33: Preparation of Compound 26R

[0318]

[0319] Compound 25R-1a (150 mg, 0.60 mmol) and diisopropylethylamine (156 mg, 1.20 mmol) were dissolved in dichloromethane (6 mL), and a diluent of acryloyl chloride (60 mg, 0.66 mmol) in dichloromethane (4 mL) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain yellow solid compound 26R-1a (190 mg, yield 100%). MS m / z 304.3 [M+H] + .

[0320] To a mixture of compound 26R-1a (190 mg, 0.63 mmol) and hydrated tin dichloride (650 mg, 3.13 mmol) in ethanol (10 mL) was added HCl aqueous solution (2.0 M, 1.57 mL), and the reaction mixture was stirred at 90 ° C for 6 hours. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure and redissolved in dichloromethane. The mixture was neutralized with saturated sodium bicarbonate aqueous solution and extracted with dichloromethane (3x 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a light yellow solid compound 26R-1b (103 mg, yield 60%). MS m / z 274.3 [M + H] + .

[0321] Compound 26R-1b (50 mg, 0.18 mmol) and compound 1k (58 mg, 0.18 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.19 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain a white solid compound 26R (16 mg, yield 16%). 1 H NMR (500 MHz, CDCl 3)δ10.97(s,1H),8.65-8.61(m,1H),8.06(s,1H),7.53-7.47(m,1H),7.31-7.24(m,1H),7.17-7.0 8(m,2H),7.00(s,1H),6.87-6.75(m,1H),6.63-6.51(m,1H),6.36(d,J=16.7Hz,1H),5.76(d,J=1 0.0Hz,1H),4.70-4.55(m,1H),4.38-4.20(m,1H),4.16-3.89(m,2H),3.55-3.45(m,1H),3.37-3. 19(m,2H),3.07(d,J=12.1Hz,1H),2.80(t,J=12.3Hz,1H),2.27(s,3H),1.85(s,3H),1.82(s,3H). MS m / z 553.5[M+H] + .

[0322] Example 34: Preparation of Compound 27R

[0323]

[0324] A mixture of compound 25R-1a (150 mg, 0.60 mmol), N-methylpiperidone (11R-1a, 340 mg, 3.01 mmol) and 2 drops of acetic acid in methanol (8 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (113 mg, 1.81 mmol) was then added and the reaction mixture was stirred overnight at room temperature. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and then extracted with dichloromethane (3 x 15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain yellow solid compound 27R-1a (219 mg) which was directly used in the next step. MS m / z 347.4 [M+H] + .

[0325] Compound 27R-1a (219 mg, 0.63 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (10 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% triethylamine) to obtain an off-white solid compound 27R-1b (182 mg, two-step yield 96%). MSm / z 317.4 [M+H] + .

[0326] Compound 27R-1b (50 mg, 0.16 mmol) and compound 1k (50 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.16 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 50: 1, 2% triethylamine) to obtain a light yellow solid compound 27R (43 mg, yield 46%). 1 H NMR (500MHz, CD 3 OD)δ8.51-8.45(m,1H),8.02(s,1H),7.60-7.50(m,2H),7.22(t,J=6.5Hz,1H),6.96(d,J=2.3Hz,1H ),6.79(d,J=2.3Hz,1H),4.53(t,J=10.6Hz,1H),3.94(dd,J=10.6,2.7Hz,1H),3.11-3.01(m,2H),3. 00-2.77(m,5H),2.69(dd,J=11.7,4.1Hz,1H),2.51-2.42(m,1H),2.26(s,3H),2.23-2.15(m,1H),2 .17(s,3H),2.05(t,J=11.0Hz,2H),1.88-1.80(m,2H),1.84(s,3H),1.82(s,3H),1.63-1.49(m,2H). MS m / z 596.6[M+H] + .

[0327] Example 35: Preparation of Compound 27S

[0328]

[0329] Compound 23S-1c (200 mg, 0.57 mmol) was dissolved in methanol (2 mL), and a methanol solution of hydrochloric acid (1 mL, 4 M) was added dropwise under an ice bath. After the addition was complete, the mixture was stirred at room temperature for 1 hour. TLC detected that the reaction was complete. The reaction mixture was concentrated under reduced pressure, and then a saturated aqueous sodium bicarbonate solution (10 mL) was added, and extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid compound 27S-1a (100 mg, yield 70%). MS m / z 250.3 [M+H] + .

[0330] A mixture of compound 27S-1a (100 mg, 0.40 mmol), N-methylpiperidone (11R-1a, 227 mg, 2.00 mmol) and 2 drops of acetic acid in methanol (5 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (75 mg, 1.21 mmol) was then added, and the reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and then extracted with dichloromethane (3x15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain yellow solid compound 27S-1b (140 mg) which was directly used in the next step. MS m / z 347.4 [M+H] + .

[0331] Compound 27S-1b (55 mg, 0.16 mmol) and palladium carbon catalyst (10%, 10 mg) were added to methanol (10 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain an off-white solid compound 27S-1c (50 mg, yield 99%). MSm / z 317.4 [M+H] + .

[0332] Compound 27S-1c (50 mg, 0.16 mmol) and compound 1k (50 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.16 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 50: 1, 2% triethylamine) to obtain a light yellow solid compound 27S (33 mg, yield 35%). 1 H NMR (500MHz, CD 3OD)δ8.51-8.45(m,1H),8.03(s,1H),7.60-7.51(m,2H),7.23(t,J=6.5Hz,1H),6.96(s,1H),6 .80(s,1H),4.55(t,J=10.8Hz,1H),3.97(dd,J=10.5,2.0Hz,1H),3.13-3.05(m,2H),3.00-2.9 4(m,3H),2.90-2.82(m,2H),2.72(dd,J=12.0,4.0Hz,1H),2.51-2.42(m,1H),2.32(s,3H),2. 26-2.12(m,3H),2.18(s,3H),1.93-1.80(m,2H),1.85(s,3H),1.82(s,3H),1.65-1.52(m,2H). MS m / z 596.6[M+H] + .

[0333] Example 36: Preparation of Compound 28R

[0334]

[0335] Compound 20R-1c (123 mg, 0.37 mmol) and palladium carbon catalyst (10%, 30 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was then separated and purified by silica gel column chromatography (dichloromethane: methanol = 15: 1) to obtain a yellow solid compound 28R-1a (73 mg, 65%). MS m / z 306.5 [M+H] + .

[0336] Compound 28R-1a (73 mg, 0.24 mmol) and compound 1k (76 mg, 0.24 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.25 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction, and the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 10: 1) to obtain yellow solid compound 28R (28 mg, yield 21%). 1 H NMR (500 MHz, CDCl 3)δ10.95(s,1H),8.67-8.58(m,1H),8.05(s,1H),7.48(t,J=7.7Hz,1H),7.30-7.24(m,1H),7.1 9-7.09(m,2H),6.99-6.88(m,2H),6.77-6.70(m,1H),4.68(dd,J=53.0,13.0Hz,1H),4.30-4.22 (m,1H),4.08-3.81(m,2H),3.71(d,J=11.8Hz,1H),3.44(t,J=5.9Hz,2H),3.34(s,3H),3.10-2 .84(m,2H),2.74-2.63(m,1H),2.57-2.41(m,3H),1.98-1.90(m,2H),1.84(s,3H),1.82(s,3H). MS m / z585.6[M+H] + .

[0337] Example 37: Preparation of Compound 29R

[0338]

[0339] Compound 29R-1a (2.0 g, 6.09 mmol), TMSN 3 (1.4 g, 12.19 mmol), 2-aminoethanol (820 mg, 13.14 mmol), copper powder (774 mg, 12.19 mmol) were dissolved in DMA (15 mL), heated to 95 ° C under nitrogen protection and stirred until the reaction was complete. 15 mL of water was added and extracted with ethyl acetate (3 x 15 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure. The crude product was purified by column chromatography to obtain a yellow oily liquid 29R-1b (600 mg, 37%). MS m / z 265.3 [M+H] + .

[0340] Compound 29R-1b (600 mg, 2.27 mmol) and DMAP (55 mg, 0.45 mmol) were dissolved in acetonitrile (10 mL), and Boc 2 O (1.49 g, 6.81 mmol), after the addition is complete, heat to 50 ° C and stir until the reaction is complete. Add 15 mL of water and extract with ethyl acetate (3 x 15 mL). The combined organic layer is dried over anhydrous sodium sulfate, filtered, and the solvent is removed by distillation under reduced pressure. The crude product is purified by column chromatography to obtain a yellow oily liquid 29R-1c (600 mg, 57%). 1 H NMR (500 MHz, CDCl 3)δ7.69(d,J=2.6Hz,1H),7.59(d,J=2.6Hz,1H),4.19–4.12(m,2H),3.67-3. 24(m,3H),2.75-2.66(m,2H),2.41-2.34(m,5H),1.47(s,9H),1.44(s,9H).

[0341] Compound 29R-1c (140 mg, 0.30 mmol) and 10% palladium carbon catalyst (32 mg, 0.03 mmol) were dissolved in methanol (3 mL), and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to obtain a light yellow solid compound 29R-1d (60 mg, 46%). MS m / z 435.6 [M+H] + .

[0342] Compound 29R-1d (60 mg, 0.14 mmol) and compound 1k (43 mg, 0.14 mmol) were dissolved in compound 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.35 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by preparative thin plate chromatography (dichloromethane: methanol = 20: 1) to obtain a light yellow solid compound 29R-1e (12 mg, yield 13%). 1 H NMR (500MHz, CD 3 OD)δ8.53-8.50(m,1H),8.01(s,1H),7.58-7.55(m,2H),7.23-7.20(m,1H),6.52-6.50(m,1 H),6.47-6.45(m,1H),4.53(t,J=11.0Hz,1H),4.01(dd,J=10.8,1.8Hz,1H),3.30-3.29(m, 1H), 3.17 (d, J = 10.5Hz, 1H), 2.99 (d, J = 12.0Hz, 1H), 2.87 (d, J = 10.5Hz, 1H), 2.77 (t, J = 11. 8Hz,1H),2.67-2.63(m,1H),2.44(t,J=10.0Hz,1H),2.34(s,3H),1.85(s,3H),1.82(s,3H).

[0343] Compound 29R-1e (12 mg, 0.023 mmol) was dissolved in 1 mL of dichloromethane, diisopropylethylamine (30 mg, 0.23 mmol) was added, and acryloyl chloride (2.1 mg, 0.023 mmol) in 1 mL of dichloromethane was added dropwise under ice bath. The reaction mixture was stirred at room temperature for 1 hour. TLC detected that the reaction was complete. The reaction solution was quenched with water and extracted with dichloromethane. The combined dichloromethane layer was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative thin layer plate to obtain compound 29R (6 mg, 45%). MS m / z 568.6 [M+H] + .

[0344] Example 38: Preparation of Compound 30R

[0345]

[0346] A mixture of compound 25R-1a (400 mg, 1.60 mmol), N-tert-butyloxycarbonyl-4-piperidone (1.6 g, 8.02 mmol) and 3 drops of acetic acid in methanol (15 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (303 mg, 4.81 mmol) was then added, and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain a yellow oily compound 30R-1a (694 mg) which was directly used in the next step. MS m / z 433.5 [M+H] + .

[0347] Compound 30R-1a (694 mg, 1.60 mmol) was dissolved in methanol (15 mL) at room temperature, and HCl methanol solution (4.0 M, 0.80 mL) was added dropwise. The mixture was stirred at 60 ° C for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, dissolved in dichloromethane, and neutralized by adding saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (3x 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain yellow solid compound 30R-1b (400 mg, two-step yield 75%).

[0348] Compound 30R-1b (150 mg, 0.45 mmol), iodomethane (106 mg, 0.68 mmol) and potassium carbonate (125 mg, 0.90 mmol) in a toluene / methanol mixture (2 mL / 1 mL) were stirred at 50 ° C for 5 hours. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain a yellow solid compound 30R-1c (115 mg, yield 71%). MS m / z 361.4 [M+H] + .

[0349] Compound 30R-1c (115 mg, 0.31 mmol) and palladium carbon catalyst (10%, 50 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 1 hour at room temperature and 1 atmosphere of hydrogen atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a brown solid compound 30R-1d (91 mg, yield 86%). MS m / z 331.5 [M+H] + .

[0350] Compound 30R-1d (91 mg, 0.28 mmol) and compound 1k (104 mg, 0.33 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.29 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a light yellow solid compound 30R (142 mg, yield 85%). 1 HNMR (500MHz, CD 3OD)δ8.51-8.45(m,1H),8.04(s,1H),7.62-7.52(m,2H),7.24(t,J=7.5Hz,1H),6.96(s,1H),6.80 (s,1H),4.56(t,J=10.6Hz,1H),3.98(dd,J=10.6,2.0Hz,1H),3.16-2.99(m,5H),2.94-2.82(m,2H ),2.73(dd,J=11.7,4.0Hz,1H),2.54-2.46(m,3H),2.30-2.22(m,1H),2.19(s,3H),2.11(t,J=11 .8Hz,2H),1.95-1.88(m,2H),1.86(s,3H),1.83(s,3H),1.64-1.52(m,2H),1.13(t,J=7.1Hz,3H). MS m / z 610.7[M+H] + .

[0351] Example 39: Preparation of Compound 30S

[0352]

[0353] A mixture of compound 27S-1a (157 mg, 0.63 mmol), N-ethyl-4-piperidone (30S-1a, 160 mg, 1.26 mmol) and 2 drops of acetic acid in methanol (5 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (118 mg, 1.89 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 25:1, 2% ammonia water) to obtain a light yellow solid compound 30S-1b (230 mg) which was directly used in the next step. MS m / z 361.5 [M+H] + .

[0354] Compound 30S-1b (230 mg, 0.53 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (10 mL) at room temperature, and the air in the system was replaced with hydrogen, and then the reaction mixture was stirred for 1 hour at room temperature and 1 atmosphere of hydrogen atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 25: 1, 2% ammonia water) to obtain a white solid compound 33S-1c (136 mg, two-step yield 64%). MS m / z 331.4 [M+H] + .

[0355] Compound 30S-1c (70 mg, 0.2 mmol) and compound 1k (67 mg, 0.2 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.2 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure. The resulting residue was dissolved in a small amount of dichloromethane, neutralized with saturated sodium bicarbonate solution, and extracted with dichloromethane (3x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a white solid compound 30S (50 mg, yield 39%). 1 H NMR (500MHz, CD 3 OD)δ8.52-8.42(m,1H),8.04(s,1H),7.63-7.49(m,2H),7.27-7.20(m,1H),6.96(d,J=2.4Hz,1H),6. 80(d,J=2.0Hz,1H),4.56(t,J=10.6Hz,1H),3.98(dd,J=10.6,2.8Hz,1H),3.18-2.98(m,5H),2.95-2. 80(m,2H),2.74(dd,J=11.7,4.2Hz,1H),2.55-2.47(m,3H),2.27(t,J=11.0Hz,1H),2.20(s,3H),2.15 -2.06(m,2H),1.95-1.88(m,2H),1.86(s,3H),1.83(s,3H),1.65-1.55(m,2H),1.13(t,J=7.2Hz,3H). MS m / z 610.6[M+H] + .

[0356] Example 40: Preparation of Compound 31R

[0357]

[0358] A mixture of compound 25R-1a (150 mg, 0.60 mmol), tetrahydropyrone (31R-1a, 301 mg, 3.01 mmol) and 2 drops of acetic acid in methanol (5 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (113 mg, 1.81 mmol) was then added, and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain a yellow solid compound 31R-1b (201 mg) which was directly used in the next step. MS m / z 334.5 [M+H] + .

[0359] Compound 31R-1b (201 mg, 0.60 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (6 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain brown solid compound 31R-1c (209 mg, yield 86%).

[0360] Compound 31R-1c (209 mg, 0.69 mmol) and compound 1k (218 mg, 0.69 mmol) were dissolved in 2-methoxyethanol (4 mL), and HCl methanol solution (2.5 M, 0.72 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a light yellow solid compound 31R (104 mg, yield 26%). 1 HNMR (500MHz, CDCl 3 )δ10.91(s,1H),8.69-8.64(m,1H),8.07(s,1H),7.51(t,J=7.9Hz,1H),7.30- 7.23(m,1H),7.14-7.06(m,2H),6.84(s,1H),6.80(s,1H),4.79-4.48(m,1H),4 .10-3.97(m,3H),3.40(t,J=11.6Hz,2H),3.30-2.72(m,6H),2.66-2.35(m,2H) ,2.25(s,3H),1.84(s,3H),1.81(s,3H),1.79-1.73(m,2H),1.66-1.56(m,2H). MS m / z 583.6[M+H] + .

[0361] Example 41: Preparation of Compound 31S

[0362]

[0363] A mixture of compound 27S-1a (100 mg, 0.40 mmol), tetrahydropyrone (31R-1a, 120 mg, 1.20 mmol) and 2 drops of acetic acid in methanol (4 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (76 mg, 1.20 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1, 2% ammonia water) to obtain a yellow solid compound 31S-1a (157 mg) which was directly used in the next step. MS m / z 334.5 [M+H] + .

[0364] Compound 31S-1a (157 mg, 0.42 mmol) and palladium carbon catalyst (10%, 60 mg) were added to methanol (4 mL) at room temperature, and the air in the system was replaced with hydrogen, and then the reaction mixture was stirred at room temperature and 1 atmosphere of hydrogen for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a yellow solid compound 31S-1b (126 mg, two-step yield 100%). MS m / z 304.5 [M+H] + .

[0365] Compound 31S-1b (50 mg, 0.16 mmol) and compound 1k (52 mg, 0.16 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.17 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the reaction to be complete. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure. The resulting residue was dissolved in a small amount of dichloromethane, neutralized with saturated sodium bicarbonate solution, and extracted with dichloromethane (3x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain white solid compound 31S (57 mg, yield 59%). 1 H NMR (500 MHz, CDCl 3)δ10.91(s,1H),8.73-8.54(m,1H),8.07(s,1H),7.51(t,J=7.9Hz,1H),7.30- 7.27(m,1H),7.13-7.08(m,2H),6.83(s,1H),6.80(s,1H),4.78-4.50(m,1H),4 .08-3.97(m,3H),3.40(t,J=11.0Hz,2H),3.32-2.77(m,6H),2.66-2.41(m,2H) ,2.25(s,3H),1.84(s,3H),1.81(s,3H),1.81-1.76(m,2H),1.68-1.59(m,2H). MS m / z 583.7[M+H] + .

[0366] Example 42: Preparation of Compound 32R

[0367]

[0368] Compound 30R-1b (150 mg, 0.45 mmol) and diisopropylethylamine (117 mg, 0.90 mmol) were dissolved in dichloromethane (3 mL) at 0°C, and acetyl chloride (53 mg, 0.68 mmol) in dichloromethane (1 mL) was slowly added dropwise. The reaction mixture was stirred at room temperature for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain yellow solid compound 32R-1a (142 mg, yield 84%). MS m / z 375.4 [M+H] + .

[0369] Compound 32R-1a (142 mg, 0.38 mmol) and palladium carbon catalyst (10%, 50 mg) were added to methanol (5 mL) at room temperature, and the reaction mixture was stirred for 1 hour at room temperature and 1 atmosphere of hydrogen atmosphere. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 15: 1) to obtain a light yellow solid compound 32R-1b (118 mg, yield 90%). MS m / z 345.5 [M+H] + .

[0370] Compound 32R-1b (60 mg, 0.17 mmol) and compound 1k (66 mg, 0.21 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.18 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a light yellow solid compound 32R (45 mg, yield 41%). 1 HNMR (500MHz, CD 3 OD)δ8.50-8.45(m,1H),8.03(s,1H),7.62-7.51(m,2H),7.23(td,J=7.5,1.3Hz,1H),6.96(d,J=2.4 Hz,1H),6.79(d,J=2.2Hz,1H),4.54(t,J=10.7Hz,1H),4.50-4.44(m,1H),4.00-3.89(m,2H),3.15- 3.04(m,3H),3.03-2.97(m,1H),2.92-2.81(m,2H),2.79-2.73(m,1H),2.72-2.64(m,1H),2.59-2.4 4(m,2H),2.18(s,3H),2.09(s,3H),1.94-1.86(m,2H),1.85(s,3H),1.82(s,3H),1.55-1.32(m,2H). MS m / z 624.7[M+H] + .

[0371] Example 43: Preparation of Compound 32S

[0372]

[0373] A mixture of compound 27S-1a (100 mg, 0.40 mmol), N-acetyl-4-piperidone (32S-1a, 170 mg, 1.20 mmol) and 2 drops of acetic acid in methanol (4 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (76 mg, 1.20 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1, 2% ammonia water) to obtain a crude yellow solid compound 32S-1b (200 mg) which was directly used in the next step. MS m / z 375.5 [M+H] + .

[0374] Compound 32S-1b (200 mg, 0.53 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (5 mL) at room temperature, the air in the system was replaced with hydrogen, and then the reaction mixture was stirred at room temperature and 1 atmosphere of hydrogen for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a yellow solid compound 32S-1c (190 mg, two-step yield 100%). MS m / z 345.5 [M+H] + .

[0375] Compound 32S-1c (50 mg, 0.15 mmol) and compound 1k (46 mg, 0.15 mmol) were dissolved in 2-methoxyethanol (2 mL), and HCl methanol solution (2.5 M, 0.15 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure. The resulting mixture was dissolved in a small amount of dichloromethane, neutralized with saturated sodium bicarbonate solution, and extracted with dichloromethane (3x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain yellow solid compound 32S (36 mg, yield 40%). 1 H NMR (500 MHz, CDCl 3 )δ10.92(s,1H),8.68-8.61(m,1H),8.06(s,1H),7.50(t,J=7.9Hz,1H),7.26-7.21(m,1H),7.1 4-7.07(m,2H),6.87(s,1H),6.80(s,1H),4.69-4.56(m,2H),3.99(dd,J=10.7,2.5Hz,1H),3.87 (d,J=14.6Hz,1H),3.27-3.20(m,1H),3.08(t,J=12.6Hz,2H),2.97-2.81(m,4H),2.66-2.46(m, 3H),2.25(s,3H),2.10(s,3H),1.84(s,3H),1.82(s,3H),1.73-1.61(m,2H),1.54-1.42(m,2H). MS m / z 624.7[M+H] + .

[0376] Example 44: Preparation of Compound 33R

[0377]

[0378] A mixture of compound 25R-1a (100 mg, 0.40 mmol), 1-cyclopropyl-4-piperidone (33R-1a, 168 mg, 1.20 mmol) and 2 drops of acetic acid in methanol (6 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (76 mg, 1.20 mmol) was then added, and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1, 2% ammonia water) to obtain yellow solid compound 33R-1b (150 mg, yield 100%). MS m / z 373.4 [M+H] + .

[0379] Compound 33R-1b (150 mg, 0.40 mmol) and palladium carbon catalyst (10%, 50 mg) were added to methanol (5 mL) at room temperature, and the reaction mixture was stirred for 2 hours at room temperature and 1 atmosphere of hydrogen. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a brown solid compound 33R-1c (116 mg, yield 84%). MS m / z 343.4 [M+H] + .

[0380] Compound 33R-1c (50 mg, 0.15 mmol) and compound 1k (55 mg, 0.18 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.15 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a light yellow solid compound 33R (70 mg, yield 77%). 1 H NMR (500MHz, CD 3OD)δ8.52-8.45(m,1H),8.04(s,1H),7.62-7.51(m,2H),7.24(t,J=7.0Hz,1H),6.96(d,J=2.1Hz,1H),6.8 0(d,J=2.1Hz,1H),4.55(t,J=10.7Hz,1H),3.97(dd,J=10.6,2.6Hz,1H),3.15-3.04(m,4H),3.00(d,J=11. 7Hz,1H),2.91-2.80(m,2H),2.73(dd,J=11.7,4.2Hz,1H),2.54-2.45(m,1H),2.32-2.21(m,3H),2.19(s,3 H),1.91-1.78(m,2H),1.85(s,3H),1.82(s,3H),1.69-1.61(m,1H),1.57-1.43(m,2H),0.55-0.39(m,4H). MS m / z 622.7[M+H] + .

[0381] Example 45: Preparation of Compound 34R

[0382]

[0383] Compound 23R (50 mg, 0.10 mmol), 1-methyl-3-azetidinone hydrochloride (34R-1a, 18 mg, 0.15 mmol), anhydrous zinc chloride (41 mg, 0.30 mmol) were dissolved in methanol (3 mL), and sodium cyanoborohydride (19 mg, 0.30 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 100 ° C and stirred for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a white solid compound 34R (21 mg, yield 37%). 1 H NMR (500 MHz, CDCl 3)δ10.90(s,1H),8.69-8.63(m,1H),8.07(s,1H),7.54-7.47(m,1H),7.30-7.23(m,1H),7.13-7. 06(m,2H),6.81(d,J=2.3Hz,1H),6.77(s,1H),4.61(t,J=10.7Hz,1H),4.01(dd,J=10.6,2.8Hz, 1H),3.74-3.65(m,2H),3.22-3.17(m,1H),3.13-3.00(m,4H),2.91-2.84(m,1H),2.73-2.63(m, 2H),2.48(s,3H),2.47-2.45(m,1H),2.24(s,3H),2.23-2.19(m,1H),1.84(s,3H),1.81(s,3H). MS m / z 568.6[M+H] + .

[0384] Example 46: Preparation of Compound 34S

[0385]

[0386] Compound 23S (38 mg, 0.08 mmol), 1-methyl-3-azetidinone hydrochloride (34R-1a, 14 mg, 0.11 mmol), anhydrous zinc chloride (31 mg, 0.23 mmol) were dissolved in methanol (3 mL), and sodium cyanoborohydride (14 mg, 0.23 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 100 ° C and stirred for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a white solid compound 34S (16 mg, yield 37%). 1 H NMR (500 MHz, CDCl 3)δ10.90(s,1H),8.70-8.62(m,1H),8.07(s,1H),7.50(t,J=7.9Hz,1H),7.31-7.27(m,1H),7.14-7. 07(m,2H),6.82(s,1H),6.77(s,1H),4.60(t,J=10.7Hz,1H),4.01(dd,J=10.6,2.8Hz,1H),3.94-3. 84(m,2H),3.33-3.13(m,4H),3.08-3.03(m,1H),2.88(td,J=11.6,2.5Hz,1H),2.73-2.64(m,2H),2 .61(s,3H),2.50(dd,J=11.5,4.2Hz,1H),2.28-2.24(m,1H),2.24(s,3H),1.84(s,3H),1.82(s,3H). MS m / z 568.7[M+H] + .

[0387] Example 47: Preparation of Compound 35S

[0388]

[0389] Compound 35S-1a (1.0 g, 5.65 mmol), 23S-1a (1.22 g, 5.65 mmol) and potassium hydroxide (950 mg, 16.94 mmol) were dissolved in dimethyl sulfoxide (15 mL), stirred at room temperature for 3 hours, and then stirred at 60 ° C for 3 hours. After the reaction was completed, the reaction mixture was cooled and poured into ice water, and stirred at room temperature for 1 hour. The mixture was then extracted with dichloromethane (3x30 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate: dichloromethane = 5:1:1) to obtain a yellow solid compound 35S-1b (1.45 g, yield 73%). MS m / z 354.4 [M+H] + .

[0390] Compound 35S-1b (1.45 g, 4.10 mmol) was dissolved in methanol (15 mL) at room temperature, and then HCl methanol solution (4.0 M, 4 mL) was added dropwise, and the mixture was stirred at 60°C for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was concentrated under reduced pressure to remove most of the methanol, and the precipitated solid was filtered and dried to obtain yellow solid compound 35S-1c (1.08 g, yield 91%). MS m / z 254.3 [M+H] + .

[0391] A mixture of compound 35S-1c (500 mg, 1.73 mmol), N-methyl-4-piperidone (11R-1a, 587 mg, 5.18 mmol) and 4 drops of acetic acid in methanol (8 mL) was stirred at room temperature for 1 hour. Sodium cyanoborohydride (326 mg, 5.18 mmol) was then added, and the reaction mixture was stirred overnight at room temperature. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1, 2% ammonia water) to obtain a yellow solid compound 35S-1d (704 mg) which was directly used in the next step. MS m / z 351.4 [M+H] + .

[0392] Compound 35S-1d (100 mg, 0.29 mmol) and palladium carbon catalyst (10%, 80 mg) were added to methanol (3 mL) at room temperature, and the air in the system was replaced with hydrogen, and then the reaction mixture was stirred at room temperature and 1 atmosphere of hydrogen for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a yellow oily compound 35S-1e (38 mg, yield 41%). MS m / z 321.5 [M+H] + .

[0393] Compound 35S-1e (38 mg, 0.12 mmol) and compound 1k (41 mg, 0.13 mmol) were dissolved in 2-methoxyethanol (1.5 mL), and HCl methanol solution (2.5 M, 0.12 mL) was added. The reaction mixture was placed in a sealed tube, heated to 120 ° C and stirred overnight. TLC monitored the reaction to be complete. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The resulting mixture was dissolved in a small amount of dichloromethane, neutralized with saturated sodium bicarbonate solution, and extracted with dichloromethane (3x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a white solid compound 35S (60 mg, yield 84%). 1 H NMR (500 MHz, CDCl 3)δ10.89(s,1H),8.61-8.56(m,1H),8.07(s,1H),7.54(t,J=7.9Hz,1H),7.31-7.27(m,1H),7.13(t,J=6 .8Hz,1H),7.07(dd,J=14.8,2.4Hz,1H),6.81(s,1H),6.75(s,1H),4.18-4.05(m,2H),3.72-3.64(m,1H ),3.25-3.18(m,1H),3.15-3.04(m,3H),2.85(dd,J=10.9,2.8Hz,1H),2.81-2.74(m,1H),2.67-2.59(m ,1H),2.50-2.42(m,1H),2.47(s,3H),2.40-2.22(m,3H),1.95-1.86(m,4H),1.84(s,3H),1.82(s,3H). MS m / z 600.7[M+H] + .

[0394] Example 48: Preparation of Compound 36R

[0395]

[0396] Compound 23R (200 mg, 0.40 mmol), 1-Boc-3-azetidinone (36R-1a, 103 mg, 0.60 mmol), anhydrous zinc chloride (164 mg, 1.20 mmol) were dissolved in methanol (10 mL), and sodium cyanoborohydride (76 mg, 1.20 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 100 ° C and stirred for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1, 2% ammonia water) to obtain a light yellow solid compound 36R-1b (258 mg) which was directly used in the next step. MS m / z 654.7 [M+H] + .

[0397] Compound 36R-1b (258 mg, 0.52 mmol) was dissolved in methanol (10 mL), and a solution of HCl in dioxane (4.0 M, 3 mL) was added. The reaction mixture was stirred at 40 °C for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The obtained residue was dissolved in a small amount of methanol, neutralized with ammonia water, and then concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 12:1, 2% ammonia water) to obtain a light yellow solid compound 36R-1c (185 mg, two-step yield 83%). MS m / z 554.6 [M+H] + .

[0398] Compound 36R-1c (70 mg, 0.13 mmol) and diisopropylethylamine (34 mg, 0.26 mmol) were dissolved in dichloromethane (10 mL), and acetyl chloride (10 mg, 0.13 mmol) in dichloromethane (1 mL) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 15 minutes. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 15:1, 2% ammonia water) to obtain white solid compound 36R (31 mg, yield 41%). 1 H NMR (500 MHz, CDCl 3 )δ11.21(s,1H),8.67-8.62(m,1H),8.01(d,J=1.2Hz,1H),7.60-7.47(m,2H),7.32-7.27(m,1H),7.1 8-7.08(m,2H),6.80(dd,J=10.7,2.2Hz,1H),4.65-4.56(m,1H),4.17-4.11(m,1H),4.07-3.91(m,3H) ,3.89-3.78(m,1H),3.23(d,J=10.3Hz,1H),3.17-3.06(m,2H),2.94-2.86(m,1H),2.83-2.70(m,2H) ,2.57-2.49(m,1H),2.32-2.25(m,1H),2.25(s,3H),1.89(d,J=1.9Hz,3H),1.85(s,3H),1.82(s,3H). MS m / z 596.5[M+H] + .

[0399] Example 49: Preparation of Compound 36S

[0400]

[0401] Compound 23S (237 mg, 0.47 mmol), 1-Boc-3-azetidinone (36R-1a, 122 mg, 0.71 mmol), anhydrous zinc chloride (194 mg, 1.42 mmol) were dissolved in methanol (15 mL), and sodium cyanoborohydride (90 mg, 1.42 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 100 ° C and stirred for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a light yellow solid compound 36S-1a (340 mg) which was directly used in the next step. MS m / z 654.7 [M+H] +.

[0402] Compound 36S-1a (340 mg, 0.47 mmol) was dissolved in methanol (15 mL), and a solution of HCl in dioxane (4.0 M, 3 mL) was added. The reaction mixture was stirred at 40 °C for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The obtained residue was dissolved in a small amount of methanol, neutralized with ammonia water, and then concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 12:1, 2% ammonia water) to obtain a light yellow solid compound 36S-1b (206 mg, two-step yield 78%). MS m / z 554.6 [M+H] + .

[0403] Compound 36S-1b (40 mg, 0.07 mmol) and diisopropylethylamine (19 mg, 0.14 mmol) were dissolved in dichloromethane (6 mL), and acetyl chloride (6 mg, 0.07 mmol) in dichloromethane (1 mL) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 15 minutes. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 15:1, 2% ammonia water) to obtain a light yellow solid compound 36S (24 mg, yield 56%). 1 HNMR (500MHz, CDCl 3 )δ11.10(s,1H),8.69-8.59(m,1H),8.03(s,1H),7.54-7.46(m,1H),7.35-7.27(m,2H),7.17-7.07 (m,2H),6.80(dd,J=10.5,2.3Hz,1H),4.65-4.56(m,1H),4.19-4.10(m,1H),4.07-3.91(m,3H),3.8 9-3.77(m,1H),3.24(d,J=11.1Hz,1H),3.19-3.04(m,2H),2.95-2.85(m,1H),2.83-2.68(m,2H),2 .59-2.47(m,1H),2.31-2.27(m,1H),2.25(s,3H),1.89(d,J=1.9Hz,3H),1.85(s,3H),1.82(s,3H). MS m / z 596.7[M+H] + .

[0404] Example 50: Preparation of Compound 37R

[0405]

[0406] Compound 36R-1c (50 mg, 0.09 mmol), acetaldehyde (6 mg, 0.14 mmol), anhydrous zinc chloride (37 mg, 0.27 mmol) were dissolved in methanol (5 mL), and sodium cyanoborohydride (17 mg, 0.27 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 70 ° C and stirred for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain white solid compound 37R (24 mg, yield 46%). 1 H NMR (500 MHz, CDCl 3 )δ10.90(s,1H),8.68-8.63(m,1H),8.07(s,1H),7.53-7.47(m,1H),7.31-7.22(m,1H),7.13-7.05( m,2H),6.81(d,J=2.2Hz,1H),6.76(s,1H),4.60(t,J=10.7Hz,1H),4.00(dd,J=10.6,2.8Hz,1H),3.8 0-3.65(m,2H),3.23-3.17(m,1H),3.16-2.96(m,4H),2.91-2.83(m,1H),2.76-2.61(m,4H),2.49(d d,J=11.4,4.1Hz,1H),2.27-2.21(m,1H),2.24(s,3H),1.84(s,3H),1.82(s,3H),1.16-1.01(m,3H). MSm / z 582.5[M+H] + .

[0407] Example 51: Preparation of Compound 37S

[0408]

[0409] Compound 36S-1b (40 mg, 0.07 mmol), acetaldehyde (5 mg, 0.11 mmol), anhydrous zinc chloride (30 mg, 0.22 mmol) were dissolved in methanol (8 mL), and sodium cyanoborohydride (14 mg, 0.22 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 70 ° C and stirred for 1 hour. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 15: 1, 2% ammonia water) to obtain a white solid compound 37S (30 mg, yield 71%). 1 H NMR (500 MHz, CDCl 3)δ10.90(s,1H),8.72-8.59(m,1H),8.07(s,1H),7.50(t,J=7.9Hz,1H),7.30-7.26(m,1H),7.13-7.05 (m,2H),6.81(d,J=2.3Hz,1H),6.76(s,1H),4.60(t,J=10.7Hz,1H),4.00(dd,J=10.6,2.8Hz,1H),3.8 2-3.61(m,2H),3.20(d,J=10.7Hz,1H),3.15-2.93(m,4H),2.92-2.82(m,1H),2.75-2.61(m,4H),2.49 (dd,J=11.5,4.3Hz,1H),2.24(s,3H),2.22-2.17(m,1H),1.84(s,3H),1.82(s,3H),1.15-1.01(m,3H). MS m / z 582.8[M+H] + .

[0410] Example 52: Preparation of Compound 38R

[0411]

[0412] Compound 23R (40 mg, 0.08 mmol), 3-oxetanone (38R-1a, 9 mg, 0.12 mmol), anhydrous zinc chloride (33 mg, 0.24 mmol) were dissolved in methanol (3 mL), and sodium cyanoborohydride (15 mg, 0.24 mmol) was added. The reaction mixture was placed in a sealed tube, heated to 100 ° C and stirred for 2 hours. TLC monitored the completion of the reaction. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a white solid compound 38R (25 mg, yield 56%). 1 H NMR (500 MHz, CDCl 3)δ11.29(s,1H),8.68-8.62(m,1H),7.99(s,1H),7.74(bs,1H),7.54-7.48(m,1H),7.32-7.27(m, 1H),7.19-7.13(m,1H),7.11(d,J=2.4Hz,1H),6.80(d,J=2.2Hz,1H),4.74-4.53(m,5H),4.05(dd ,J=10.7,2.8Hz,1H),3.53-3.44(m,1H),3.26-3.20(m,1H),3.11-3.05(m,1H),2.98-2.91(m,1H) ,2.76-2.67(m,2H),2.54-2.49(m,1H),2.31-2.21(m,1H),2.25(s,3H),1.85(s,3H),1.83(s,3H). MS m / z 555.4[M+H] + .

[0413] Example 53: Kinase activity inhibition experiment

[0414] 1.EGFR (C797S) kinase activity inhibition experiment:

[0415] The Lantha screen Assay method was used to test the inhibitory activity of the compounds at a kinase EGFR (C797S) concentration of 10 nM and an ATP concentration of 0.1 μM, and the control substance was Staurosporine.

[0416] Experimental steps:

[0417] 1) Buffer preparation: 50 mM HEPES, pH 7.5, 0.0015% Brij-35.

[0418] 2) Preparation of reference substance Staurosporine and test samples: Staurosporine and the compounds of the present invention were prepared into gradient concentration solutions in 100% DMSO, and diluted to 10% DMSO with the above buffer, and added to a 384-well plate. For example, if the initial concentration of the compound is 10 μM, it is prepared to 1000 μM with 100% DMSO, and diluted to 10 concentration gradients, and then added to a 384-well plate. 100 nL was transferred into a 384-well plate using LIQUID HANDLE RS (LABCYTE, USA).

[0419] 3) Dilute kinase EGFR (C797S) to the required concentration using the following buffer: 50 mM HEPES, pH 7.5, 0.0015% Brij-35, 2 mM DTT. Transfer 5 μL to a 384-well plate and incubate with the compound for 10-15 minutes.

[0420] 4) Dilute the substrate to the desired concentration using the following buffer: 50 mM HEPES, pH 7.5, 0.0015% Brij-35, 10 mM MgCl 2 . Transfer 5 μL to a 384-well plate to initiate the reaction and incubate at room temperature for 30 minutes.

[0421] The reaction concentrations of each reagent in the test experiment are shown in Table 1 below.

[0422] Table 1.

[0423]

[0424] 5) Prepare 2x detection solution with TR-Fret dilution buffer, with final concentration of Antibody at 2 nM and final concentration of EDTA at 10 mM. Transfer 10 μL of detection solution to the reaction wells of a 384-well plate, shake to mix, and let stand at room temperature for 60 min.

[0425] 6) Read the excitation at 340 nm on Envision, the ratio of emission at 520 nm to 495 nm, and calculate the inhibition rate.

[0426] 7) Fitting IC using XL-fit software 50 .

[0427] The activities of representative compounds are shown in Table 2. 50 Values ​​are represented by:

[0428] A:1nM <IC 50 Value ≤50nM; B:50nM <IC 50 Value ≤ 250nM; C: 250nM <IC 50 Value ≤1000nM; D:

[0429] IC 50 Value>1000nM.

[0430] Table 2. EGFR (C797S) kinase activity inhibition (IC 50 value)

[0431] Compound EGFR(C797S) 1 A 1R A 1S A 2R B 3R A 4R A 4S A 5R B 6R B 6S A 7R B 8R A 9R C 10R A 11R A 12R B 12S B 13R B 14R B 15R B 16R B

[0432] 2. ALK kinase activity inhibition experiment:

[0433] Method 1: Caliper mobility shift assay was used to measure ALK protein kinase activity. The compound was dissolved in DMSO and diluted with kinase buffer: 50 mM HEPES, pH 7.5, 0.0015% Brij-35, 10 mM MgCl 2 , 2mM DTT; add 5μL of 5 times the final concentration of the compound (10% DMSO) to the 384-well plate. Add 10μL of 2.5 times enzyme solution and incubate at room temperature for 10 minutes, then add 10μL of 2.5 times substrate (Peptide FAM-P22 and ATP) solution. After incubation at 28°C for 60 minutes, add 25μL of stop solution (100mM HEPES at pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50mM EDTA) to terminate the reaction. Read the conversion rate data on Caliper EZReader II (Caliper Life Sciences). Convert the conversion rate to inhibition rate data (% inhibition rate = (max-conversion rate) / (max-min)*100). Where max refers to the conversion rate of the DMSO control, and min refers to the conversion rate of the enzyme-free control. The curve was drawn with the compound concentration and inhibition rate as the horizontal and vertical coordinates, and the XLFit excel add-inversion 5.4.0.8 software was used to fit the curve and calculate the IC 50 .

[0434] Method 2: Caliper mobility shift assay was used to measure ALK protein kinase activity. The method was basically the same as method 1, with some parameters adjusted. The starting concentration of the compound was 1 mM, and it was diluted 4-fold to 6 (7 for some compounds). Use the dispenser Echo 550 to transfer 250 nL of 100-fold final concentration of the compound to the target plate 3573, add 10 μL of 1.25 nM ALK at a final concentration, and incubate at room temperature for 10 minutes (negative control wells contain 10 μL kinase buffer and 250 nL 100% DMSO; positive control wells contain 10 μL kinase solution and 250 nL 100% DMSO). On the kinase ALK, add 15 μL of a mixed solution of ATP with a final concentration of 30 μM and 3 μM substrate peptide 22 for 25 minutes; add 30 μL of EDTA-containing stop detection solution to stop the kinase reaction. Use Caliper EZ Reader to read the conversion rate. Conversion inhibition rate % = (positive control conversion rate mean % - sample conversion rate % / (positive control conversion rate mean % - negative control conversion rate mean %). Wherein: negative control wells represent the conversion rate readings of wells without enzyme activity; positive control wells represent the conversion rate readings of wells without compound inhibition. The log value of the concentration is used as the X-axis, and the percentage inhibition rate is used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism 5 is used to fit the dose-effect curve to obtain the IC value of each compound on the enzyme activity. 50 Value. Calculation formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)).

[0435] The activities of representative compounds are shown in Table 3. 50 Values ​​are represented by:

[0436] A:1nM <IC 50 Value ≤50nM; B:50nM <IC 50 Value ≤ 250nM; C: 250nM <IC 50 Value ≤1000nM; D:

[0437] IC 50 Value>1000nM.

[0438] Table 3. ALK kinase activity inhibition (IC 50 value)

[0439]

[0440]

[0441] Note: Compounds 1-16R were tested using method 1, and compounds 25R-32R were tested using method 2.

[0442] 3.EGFR (T790M / L858R / C797S) kinase activity inhibition experiment:

[0443] The Caliper mobility shift assay was used to detect the inhibitory effect of the compounds on the activity of EGFR (T790M / L858R / C797S) kinase. The basic method is the same as the ALK activity test method 2. The compound test concentration is 2μM, and 5-fold dilution is made into 6 (7 for some compounds) concentration points. Use the dispenser Echo 550 to transfer 250nL of the compound at 100 times the final concentration to the target plate 3573, add 10μL of EGFR (T790M / L858R / C797S) kinase solution with a final concentration of 2.5nM, and incubate at room temperature for 10 minutes (negative control wells contain 10μL kinase buffer and 250nL 100% DMSO; positive control wells contain 10μL kinase solution and 250nL 100% DMSO). Add 15 μL of a mixed solution of ATP with a final concentration of 40 μM and 3 μM substrate peptide No. 22 and react for 60 minutes. Add 30 μL of EDTA-containing termination detection solution to stop the kinase reaction. Use Caliper EZ Reader to read the conversion rate. Conversion inhibition rate % = (positive control conversion rate mean % - sample conversion rate % / (positive control conversion rate mean % - negative control conversion rate mean %). Among them: negative control wells represent the conversion rate readings of wells with no enzyme activity; positive control wells represent the conversion rate readings of wells with no compound inhibition. With the log value of the concentration as the X-axis and the percentage inhibition rate as the Y-axis, the log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism 5 was used to fit the dose-effect curve to obtain the IC of each compound on the enzyme activity. 50 Value. Calculation formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)).

[0444] The activities of representative compounds are shown in Table 4. 50 Values ​​are represented by:

[0445] A:IC 50 Value ≤3nM; B:3nM <IC 50 Value ≤15nM; C:15nM <IC 50Value ≤100nM; D:IC 50 Value > 100

[0446] nM.

[0447] Table 4. EGFR (T790M / L858R / C797S) kinase activity inhibition (IC 50 value)

[0448]

[0449]

[0450] 4. MAP4K1 (HPK1) kinase activity inhibition experiment:

[0451] The Caliper mobility shift assay was used to detect the inhibitory effect of compounds on the activity of MAP4K1 (HPK1) kinase. The starting concentration of the compound test was 500nM, 5-fold dilution, and 5 concentration points. Use the dispenser Echo 550 to transfer 250nL of 100-fold final concentration of the compound to the target plate 3573, add 10μL of kinase solution with a final concentration of 1.25Nm MAP4K1 (HPK1), and incubate at room temperature for 10 minutes (negative control wells contain 10μL kinase buffer and 250nL 100% DMSO; positive control wells contain 10μL kinase solution and 250nL 100% DMSO). On the kinase MAP4K1 (HPK1), add 15μL of a mixed solution of ATP with a final concentration of 10.1μM and 3μM substrate peptide 25 for 120 minutes. Add 30μL of EDTA-containing stop detection solution to stop the kinase reaction. The conversion rate was read using Caliper EZ Reader. The conversion inhibition rate % = (positive control conversion rate mean % - sample conversion rate % / (positive control conversion rate mean % - negative control conversion rate mean %). Among them: negative control wells represent the conversion rate readings of wells without enzyme activity; positive control wells represent the conversion rate readings of wells without compound inhibition. The log value of the concentration was used as the X-axis, and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphPadPrism 5 was used to fit the dose-effect curve, thereby obtaining the IC value of each compound on the enzyme activity. 50 Value. Calculation formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)).

[0452] The activities of representative compounds are shown in Table 5. 50Values ​​are represented by:

[0453] A:IC 50 Value ≤1nM; B:1nM <IC 50 Value ≤10nM; C:10nM <IC 50 Value ≤100nM; D:IC 50 Value > 100

[0454] nM.

[0455] Table 5. MAP4K1 (HPK1) kinase activity inhibition (IC 50 value)

[0456]

[0457]

[0458] Example 54: Ba / F3_EGFR (del19-T790M-C797S) cell anti-proliferation experiment

[0459] Method 1: Cell culture: Ba / F3_EGFR del19 / T790M / C797S cell culture medium is RPMI-1640+10% FBS+1% double antibody. Cells are placed at 37°C, 5% CO 2 Culture in an incubator.

[0460] Cell plating and compound treatment: 1) Culture cells routinely until the cell saturation is 80%-90%. When the number reaches the requirement, collect the cells. 2) Resuspend with the corresponding fresh culture medium, take a small amount of cells to count, and prepare a cell suspension of appropriate density. 3) Inoculate the cell suspension into a 384-well plate at 700 cells / well, 30μL per well. 4) Use Echo to add the compound to the corresponding cell well. The highest starting concentration of the compound is 1-10μM, 3-fold dilution, 8 concentrations, blank control wells are cells plus 0.1% DMSO, and positive control wells are cells plus 10μM Brigatinib. Cells are incubated at 37°C, 5% CO 2 Culture in an incubator for 72 h.

[0461] CTG method detection: 1) Add 30 μL CTG reagent (CelltiterGlo kit) to each well and incubate at 37°C, 5% CO, protected from light. 2 2) Read the chemiluminescent signal using Envision instrument.

[0462] Data analysis: IC 50 The results were analyzed by GraphPad Prism 6 software. The IC of the compound was obtained using the following nonlinear fitting formula: 50(half inhibitory concentration): Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 =(blank control well reading-compound well reading) / (blank control well reading-positive control well reading)×100.

[0463] Method 2: Cell plating and compound treatment: 1) Take Ba / F3_EGFR Del19 / T790M / C797S cells, centrifuge at 800rpm for 5min, discard the supernatant, resuspend in fresh culture medium (RPMI-1640 + 10% FBS), take out a small amount of cells and count them with ViCell. 2) Adjust the cell density and inoculate 2000 cells / well in a 384-well plate at 37°C, 5% CO 2 Incubate in the incubator for 4 hours. 3) Set the Tecan HP D300 program and add the compound to the well plate. The highest starting concentration of the compound is 1-4 μM, 3-fold dilution, 9 concentrations, and the DMSO content in each well is unified to 0.2%. The cells are incubated at 37°C, 5% CO 2 Culture in an incubator for 72 h.

[0464] CTG method detection: 1) Take out the CTG reagent and cell plate and place them at room temperature for 30 minutes, add 25 μL CTG to each well, shake at medium speed for 2 minutes, centrifuge at 1000 rpm for 1 minute, and let stand at room temperature for 10 minutes. 2) Envision instrument reads the chemiluminescent signal value.

[0465] Data analysis: IC 50 The results were analyzed by XL-fit 5.0 software from IDBS.

[0466] The activities of representative compounds are shown in Table 6. 50 Values ​​are represented by:

[0467] A:IC 50 Value ≤20nM; B:20nM <IC 50 Value ≤ 100nM; C: 100nM <IC 50 Value ≤200nM; D:IC 50 value

[0468] >200nM.

[0469] Table 6. Inhibition of antiproliferative activity of BaF3_EGFR (del19-T790M-C797S) cells (IC 50 value)

[0470]

[0471]

[0472] Example 55: Pharmacokinetic study in rats

[0473] Instrument: XEVO TQ-S LC-MS instrument produced by Waters. All measured data were collected and processed by Masslynx V4.1 software, and calculated and processed by Microsoft Excel. WinNonLin 8.0 software was used to calculate pharmacokinetic parameters using the statistical moment method. The main parameters included were kinetic parameters T max , T 1 / 2 , C max , AUC last etc. Chromatographic column: ACQUITYUPLC BEH C18 (2.1mm×50mm, 1.7μm); column temperature 40℃; mobile phase A is water (0.1% formic acid), mobile phase B is acetonitrile, flow rate is 0.350ml / min, gradient elution is adopted, elution gradient is 0.50min: 10%B; 1.50min: 10%B; 2.30min: 95%B; 2.31min: 10%B; 3.00min: stop. Injection volume: 5μL.

[0474] Animals: 3 SD male rats, weighing 200-220g, were purchased and raised in the laboratory of the Experimental Animal Center for 2 days before use. They were fasted for 12 hours before administration and within 4 hours after administration, and had free access to water during the experiment. Blood samples were collected at the prescribed time after oral gavage.

[0475] Solvent: 0.5% Methylcellulose (aqueous solution containing 0.4% Tween 80 and 1% ethanol). Preparation of intragastric administration solution: accurately weigh the compound, add it to the solvent, and ultrasonicate it at room temperature for 5 minutes to completely dissolve the drug, and prepare a 0.3 mg / ml drug solution.

[0476] Drug samples: Representative compounds of the structure shown in the patent formula (I) of the present invention are generally taken in multiple samples with similar structures (molecular weight difference is more than 2 units), accurately weighed, and administered together (cassette PK). In this way, multiple compounds can be screened at the same time and their oral absorption rates can be compared. Single administration is also used to study the pharmacokinetics of drug samples in rats.

[0477] Blood was collected from the orbit at 0.25, 0.5, 1, 2, 4, 9, 12, and 24 hours after intragastric administration, and the supernatant plasma was obtained for LC-MS / MS analysis after centrifugation in a sodium heparin-treated plastic centrifuge tube.

[0478] The compound is accurately weighed and prepared into different concentrations, and quantitative analysis is performed on the mass spectrometer to establish a standard curve, and then the concentration of the compound in the above plasma is tested to obtain the concentration of the compound at different time points. All the measurement data are collected and processed by relevant software, and the pharmacokinetic parameters are calculated using the statistical moment method (mainly including the kinetic parameter T max , T 1 / 2 , C max , AUC last etc.). The results are as follows:

[0479] Table 7. Pharmacokinetic parameters of the compounds after oral administration in SD rats

[0480] Compound dose <![CDATA[T 1 / 2 (h)]]> <![CDATA[T max (h)]]> <![CDATA[C max (ng / mL)]]> <![CDATA[AUC 0-24h (mg / mL)]]> 1R 4mg / kg 12.67 1.08 23.10 101.89 25R 3mg / kg 6.75 6.33 61.44 585.35 27S 3mg / kg 5.20 10.00 140.33 2102.82 27R 3mg / kg 5.17 5.33 195.84 2751.90 31R 3mg / kg 5.18 6.33 78.37 686.57 31S 3mg / kg 6.27 1.00 160.17 1345.22 32R 3mg / kg 1.95 2.00 306.38 3419.6 32S 3mg / kg 2.22 3.33 351.99 3263.69 33R 3mg / kg 3.88 7.33 120.36 1775.84 35S 3mg / kg 3.97 4.33 15.03 179.27

[0481] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound having a structure as shown in the following formula (I), or an optical isomer or a pharmaceutically acceptable salt thereof: In formula (I): "*" indicates chiral center; Each R is independently C 1-4 alkyl; Each R 1 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, or CN; Each R 2 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, or CN; Each R 3 Each is independently hydrogen, deuterium, or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atoms to which it is attached may optionally together form a carbonyl group (C=O); Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, OR h , SR h NR h R h 、CN、C(O)R e 、C(O)OR h , C(O)NR h R h 、OC(O)R e , or NR h C(O)R e ; J is O; G is NR f or O; m is 0, 1, 2, 3, or 4; n is 0, 1, 2, or 3; p is 0, 1, or 2; q is 0, 1, 2, or 3; R f For hydrogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O)2R e , or S(O)2NR h R h ;in, Each of the above groups is unsubstituted or substituted with 1-3 R e replace; Each R e Each is independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Halogenated alkenyl, C 1-4 Alkoxy substituted C 2-4 Alkenyl, hydroxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkenyl, C 3-8 Cycloalkyl substituted C 2-4 alkenyl, 3- to 8-membered heterocyclic substituted C 2-4 Alkenyl, aryl substituted C 2-4 Alkenyl, heteroaryl substituted C 2-4 Alkenyl, C 2-4 Alkynyl, C 2-4 Haloalkynyl, C 1-4 Alkoxy substituted C 2-4 Alkynyl, hydroxy substituted C 2-4 Alkynyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkynyl, C 3-8 Cycloalkyl substituted C 2-4 Alkynyl, 3- to 8-membered heterocyclic substituted C 2-4 Alkynyl, C 3-8 Cycloalkyl, or 3- to 8-membered heterocyclic group; Each R h are independently hydrogen, or C 1-4 Alkyl; or two R h Together with the nitrogen atom to which it is attached, it forms a 3- to 8-membered heterocyclic group containing 1 or 2 N atoms and 0 or 1 heteroatom selected from O and S; wherein each of the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h 、C(O)NR h R h , provided that the chemical structure formed is stable and meaningful; e and R h is defined as above; Unless otherwise specified, the above-mentioned aryl group is an aromatic group containing 6 to 12 carbon atoms; the heteroaryl group is a 5- to 15-membered heteroaromatic group.

2. The compound according to claim 1, or an optical isomer or a pharmaceutically acceptable salt thereof, characterized in that: Formula (I) is: The definitions of the various groups are as described in claim 1.

3. The compound according to claim 2, characterized in that Each R is independently C 1-2 alkyl; Each R 1 are independently hydrogen, deuterium, halogen, or C 1-2 alkyl; Each R 2 are independently hydrogen, deuterium, halogen, or C 1-2 alkyl; Each R 3 Each is independently hydrogen or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atom to which it is attached together forms a carbonyl group (C=O); Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, NR h R h , or NR h C(O)R e ; m is 0, 1, or 2; n is 0, 1, or 2; p is 0, 1, or 2; q is 0, 1, or 2; Where R e and R h The definition is as stated in claim 1.

4. The compound according to claim 3, characterized in that Formula (I) is: Where R 2 is F, Cl, or Br; each R 3 Each is independently hydrogen or C 1-4 Alkyl; or when two R 3 When these two R 3 The carbon atom connected to it together forms a carbonyl group (C=O); each R 4 Each is independently hydrogen, deuterium, halogen, C 1-4 Alkyl, NR h R h , or NR h C(O)R e ; n is 0, 1, or 2; q is 0, 1, or 2; wherein J, G, R e , and R h The definition is as stated in claim 1.

5. The compound according to claim 4, characterized in that The structural fragment in formula (IIIa) Selected from: Represents the connection site between the above structural fragment and other structures in formula (IIIa); Among them, each R 3 Each is independently hydrogen or C 1-4 Alkyl; when two R 3 When attached to the same carbon atom, the two R 3 and the carbon atoms connecting them can together form C=O; Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ; n is 0, 1, or 2; q is 0 or 1; R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O)2R e , or S(O)2NR h R h wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition is as stated in claim 1.

6. The compound according to claim 4, characterized in that The structural fragment in formula (IIIa) Selected from: Represents the connection site between the above structural fragment and other structures in formula (IIIa); Among them, each R 3 Each is independently hydrogen or C 1-4 Alkyl; when two R 3 When attached to the same carbon atom, the two R 3 and the carbon atoms connecting them can together form C=O; Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ; n is 0, 1, or 2; q is 0 or 1; R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O)2R e , or S(O)2NR h R h wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition is as stated in claim 1.

7. The compound according to claim 5 or 6, characterized in that Formula (I) is: Among them, each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ;q is 0 or 1; R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e 、C(O)OR h 、C(O)NR h R h 、S(O)2R e , or S(O)2NR h R h wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition is as stated in claim 1.

8. The compound according to claim 7, characterized in that The R f Selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O)2R e wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition of is as above.

9. The compound according to claim 7, characterized in that Formula (I) is: Among them, R 4 For hydrogen, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ; R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O)2R e wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition is as stated in claim 1.

10. The compound according to any one of claims 2 to 4, characterized in that Formula (I) is: Among them, R 4 For hydrogen, halogen, C 1-2 Alkyl, NR h R h , or NR h C(O)R e ; R f For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, aryl, heteroaryl, C(O)R e , or S(O)2R e wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally and independently substituted by 1-3 substituents each independently selected from the group consisting of halogen, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group, CN, NO2, OR h , SR h NR h R h 、C(O)R e 、C(O)OR h , or C(O)NR h R h ; where R e and R h The definition is as stated in claim 1.

11. The compound according to claim 1, characterized in that Formula (I) is: Among them, R 4 For hydrogen, halogen, C 1-2 alkyl; s and t are each independently 1, 2, or 3; A is NR k , or O,; where R k For hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, hydroxy substituted C 1-4 Alkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, di(C 1-4 Alkyl)amino-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl, 3- to 9-membered heterocyclyl, C(O)R e 、C(O)OR h , or C(O)NR h R h ; each R e are independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 Alkenyl, C 1-4 Alkoxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-6 Cycloalkyl, or 3- to 8-membered heterocyclic group; R g and R h The definition is as stated in claim 1.

12. The compound according to claim 1, characterized in that The various R e Each independently selected from the group consisting of hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy substituted C 1-4 Alkyl, C 2-4 Alkenyl, C 1-4 Alkoxy substituted C 2-4 Alkenyl, hydroxy substituted C 2-4 Alkenyl, di(C 1-4 Alkyl)amino-substituted C 2-4 alkenyl, 3- to 6-membered heterocyclic substituted C 2-4 Alkenyl, C 2-4 Alkynyl, C 3-8 Cycloalkyl, 3- to 8-membered heterocyclic group.

13. The compound according to claim 1, characterized in that Each R 4 Each is independently hydrogen, deuterium, halogen, C 1-2 Alkyl, or NHC(O)CH=CH2.

14. A compound selected from the group consisting of: in, "*" indicates a chiral center. Unless R or S is indicated, a compound with "*" indicates that it can be a racemate, an R configuration, or an S configuration.

15. The use of the compound of formula (I), its optical isomer or pharmaceutically acceptable salt as claimed in claim 1, characterized in that: Used for: (a) preparing a drug for treating a disease associated with protein kinase activity or expression; (b) preparing protein kinase targeted inhibitors; and / or (c) non-therapeutic inhibition of protein kinase activity in vitro; Wherein, the protein kinase is selected from the following group: EGFR, ALK, HPK1, or a combination thereof.

16. The use according to claim 15, characterized in that The protein kinase is EGFR (C797S).

17. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (i) an effective amount of the compound of formula I according to claim 1, or an optical isomer or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

18. A method for preparing a compound of formula (I), characterized in that: The compound has a structure shown in IIIf or IIIg, and the method comprises the steps of: Reacting a compound of formula 4-D1 with a compound of formula 1-A2 to obtain a compound of formula 4-D2-1 or 4-D2-2; In the presence of a palladium catalyst, reacting a compound of formula 4-D2-1 or 4-D2-2 with Me4Sn to obtain a compound of formula 4-D3-1 or 4-D3-2; Performing a reduction reaction with a compound of formula 4-D3-1 or formula 4-D3-2 to obtain a compound of formula 4-D4-1 or formula 4-D4-2; Reacting a compound of formula Ib with a compound of formula 4-D4-1 or a compound of formula 4-D4-2 to obtain a compound of formula IIIf or IIIg; Wherein, the definition of each group is as described in any one of claims 1-14.

19. A method for preparing a compound of formula (I), characterized in that: The compound has a structure shown in IIIi, and the method comprises the steps of: Reacting a compound of formula 5-E2 with a compound of formula Ib to obtain a compound of formula IIIh; The compound of formula IIIh is subjected to a reductive amination reaction with the compound of formula 5-E3 to obtain a compound of formula IIIi; Wherein, the definition of each group is as described in any one of claims 1-14.

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