Monocyclic aryl substituted CDK12 / 13-containing degradation agent as well as preparation method, pharmaceutical composition and application of monocyclic aryl substituted CDK12 / 13-containing degradation agent

Abstract

The invention relates to a degradation agent containing monocyclic aryl substituted CDK12 / 13 as well as a preparation method, a pharmaceutical composition and application of the degradation agent. The degradation agent for cell cycle-dependent protein kinase 12 / 13 (CDK12 / 13) has a structure as shown in a formula (I), and the compound can be used as a protein kinase degradation agent, can effectively degrade CDK12 / 13 protein with high selectivity, and can inhibit proliferation, migration and invasion of various tumor cells. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of chemical medicine, and particularly relates to a degrader of cyclin-dependent protein kinase 12 / 13 (CDK12 / 13), a pharmaceutical composition thereof, and applications thereof. Background Art

[0002] Protein kinases are key regulatory molecules of cell functions and constitute one of the largest and most functionally diverse gene families. By phosphorylating substrate proteins, protein kinases direct the activities, localizations, and overall functions of many proteins and are involved in almost all cellular activities. Abnormalities in the expression, activation, localization, etc. of protein kinases are closely related to the occurrence and development of various diseases and are important driving factors for various diseases such as tumors and inflammation. As of December 31, 2020, the FDA has approved a total of 62 kinase inhibitor drugs for market. Research has found that the non-kinase functions of proteins play important roles in diseases such as tumors. The non-kinase functions of kinases such as CDK6, FAK, EGFR, PIPK3, PDK1, BRAF, CRAF, CHK2, ZAP70, AKT, and Aurora A have been reported in the literature. Therefore, simply inhibiting the activity of kinases cannot completely inhibit the non-kinase functions of kinases, which may bring potential adverse effects such as low efficacy and drug resistance. Therefore, developing protein degraders is expected to comprehensively inhibit the enzymatic activity and non-kinase functions of kinases and exert a potent therapeutic effect. Protein degraders based on the PROTAC principle are currently the most successful and mature protein degradation strategies and have been successfully used for the development of degraders for multiple kinases and other targets.

[0003] PROTAC specifically refers to a class of small molecule compounds that can specifically recognize and induce the degradation of target proteins; the molecular structure consists of three parts: a target protein recognition ligand, a Linker, and an E3 recognition ligand. The advantage of PROTAC is that it can not only effectively inhibit the kinase activity of target proteins but also rapidly degrade and remove target proteins. In theory, only a catalytic amount of the drug is required to degrade almost all proteins (including membrane proteins) in cells, so it has high safety, drug resistance, and broad application prospects. Currently, degraders for target proteins such as ERR, ABL, BET, and CDK4 / 6 have been successfully developed, and the results show that they can not only treat the proliferation of gene-driven tumors but also overcome inhibitor resistance. In 2018, Pfizer announced an investment of $830 million in the research and development of protein degrader drugs based on PROTAC technology. In March 2019, Arinas, a partner of WuXi AppTec, announced that its developed protein degrader ARV-110 for androgen receptor entered clinical research; this is the world's first protein degrader to enter the clinical research stage.

[0004] CDK12 / 13 (Cyclin-dependent kinase 12 / 13) belongs to the cyclin-dependent kinase (CDK) family of serine / threonine protein kinases and forms a complex with Cyclin K to exert its biological functions. CDK12 and CDK13 contain 1,490 and 1,512 amino acids respectively and share 46% homology. The kinase domain consists of 300 amino acids with a homology as high as 92%. CDK12 / 13 forms a complex with Cyclin K to phosphorylate the C-terminal domain (CTD) of RNA polymerase II (RNA PolII). The CTD consists of a highly repetitive sequence of seven amino acids, YSPTSPS. In humans, the CTD contains a 52-repeat unit. CDK12 / 13 mainly phosphorylates Ser2 to regulate transcription and post-transcriptional mRNA processing. Genetic studies have shown that CDK12 promotes the production of full-length gene transcripts by inhibiting the cleavage of intronic polyadenylation sites. Many homologous recombination repair genes (such as BRCA1 / 2, ATM, ATR, FANCD2, FANCI, etc.) contain more intronic polyadenylation sites. Therefore, the expression of these genes is more sensitive to the loss or inhibition of CDK12. Compared with other transcription CDKs, the N-terminus of CDK12 / 13 contains an additional arginine / serine-rich (RS) motif, which is commonly found in proteins involved in pre-mRNA splicing. Proline-rich motifs (PRIM) are also found in the N- and C-termini and may serve as binding sites for proteins containing SH3, WW, or profilin domains. These structures suggest that CDK12 / 13 may have non-kinase functions (such as protein-protein interactions) that play important roles in mRNA splicing and mRNA 3'-end processing. Therefore, using only kinase inhibitors of CDK12 / 13 as probe tool molecules to study the biological functions of CDK12 / 13 is far from sufficient. Inducing the degradation of CDK12 / 13 using proteolysis-targeting chimeras (PROTACs), while inhibiting both its kinase and non-kinase functions, has great potential in studying the biological functions of CDK12 / 13 and disease treatment and has attracted increasing interest from scientists in recent years. Summary of the Invention

[0005] Based on this, the present invention provides a class of trans-1,4-cyclohexanediamine compounds and their use as CDK12 / 13 degrading agents. These compounds can effectively and highly selectively degrade the CDK12 / 13 protein kinase and inhibit the proliferation, migration, and invasion of various tumor cells.

[0006] In a first aspect of the present invention, there is provided a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a prodrug molecule thereof:

[0007]

[0008] Wherein, Z is CH 2 or CO;

[0009] V is selected from the group consisting of:

[0010] X and Y are each independently selected from the group consisting of: N, CH or CR 3 ; wherein, R 3 is selected from the group consisting of: halogen, cyano, hydroxy, amino, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 3 alkoxy, halo-C 1 -C 3 alkoxy, C 3 -C 8 cycloalkyl, or a 3- to 8-membered heterocyclic group;

[0011] B is selected from the group consisting of: NH, O, CO, CH 2 ; wherein, each U and W is independently selected from the group consisting of: N or CH; each m, p, m', p' is independently: 0, 1, 2 or 3;

[0012] Ring A is selected from the group consisting of:

[0013] Wherein, Q and W are each independently selected from: CH, N;

[0014] R' is independently selected from: hydrogen, halogen, cyano, hydroxy, substituted hydroxy, amino, substituted amino, C 1 -C 5 alkyl, halo-C 1 -C 5 alkyl, C 1 -C 5 alkoxy, halo-C 1 -C 5 alkoxy, C 3 -C 8 cycloalkyl, and when Q and W are independently selected from CH, R' can be a substituent on Q and W;

[0015] D, E, F, G are each independently selected from the group consisting of: CH, N, CR6 ; wherein, R 6 is selected from the group consisting of: halogen, trifluoromethyl, hydroxy, cyano, amino, methyl, methoxy, trifluoromethoxy;

[0016] R 1 is selected from the group consisting of: H, -NHR 7 , -OR 7 , -(C(R 9 )R 8 )R 7 ;

[0017] wherein, R 7 is -R 10 , -CH 2 R 10 or -(CH 2 ) 2 R 10 ;

[0018] R 8 , R 9 are each independently selected from the group consisting of: hydrogen, halogen, cyano, methyl, halomethyl, methoxy, halomethoxy, ethyl, haloethyl, ethoxy, haloethoxy, hydroxy, amino, 3-8 membered heterocycles containing 1, 2 or 3 heteroatoms selected from O, S or N;

[0019] Or R 8 , R 9 and the C atom to which they are attached together form a 3-7 membered heterocycle;

[0020] R 10 is selected from the group consisting of:

[0021] 1) cyano, C 1 -C 5 alkyl, halocarbonyl 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 3 ~C 10 cycloalkyl, substituted or unsubstituted 5-12 membered aromatic ring, substituted or unsubstituted 3-12 membered heterocycle;

[0022] 2)

[0023] wherein Q 1 , Q 2 , Q 3 , Q 4 , Q 5 are each independently selected from: CH, N or CR 11 ;

[0024] Each R11 Each independently selected from the following group: halogen, cyano, hydroxy, amino, nitro, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 4 alkoxy, halo-C 1 -C 4 alkoxy, C 3 -C 8 cycloalkyl;

[0025] R 2 selected from the following group: H, C 1 -C 3 alkyl,

[0026] When B is selected from the following group: NH, O, CO, CH 2 ; wherein each U and W is independently selected from the following group: N or CH; each m, p, m', p' is independently: 0, 1, 2 or 3; when

[0027] Linker is:

[0028] wherein,

[0029] R L1 、R L2 、R L3 、R L4 and R L5 are the same or different and are each independently selected from the following group of substituted or unsubstituted groups: chemical bond, CH 2 、CHD、CD 2 、C=O、O、NH、SO、SO 2 、P=O、NHCO、NHSO 2 、OCH 2 、OCH 2 CH 2 、CH 2 OCH 2 、NHCH 2 、NMeCH 2 、NHCH 2 CH 2 、NMeCH 2 CH 2 、CH 2 NHCO、NHCOCH 2 、

[0030]

[0031] R L6 is a ring and is optionally selected from the following structures:

[0032]

[0033]

[0034]

[0035] wherein each n is independently 0, 1, 2, 3, 4, 5 or 6; each r and m are independently 0, 1 or 2; each U and W are independently selected from the group consisting of: N or CH

[0036] p L1 , p L2 , p L3 , p L4 , p L5 and p L6 are independently selected from 0, 1, 2, 3, 4, 5, or 6.

[0037] When B is wherein each m, p, m', p' are independently: 0, 1, 2 or 3; then

[0038] Linker is:

[0039] wherein,

[0040] R L1 , R L2 , R L3 , R L4 , R L5 and R L6 are the same or different and are each independently selected from the group consisting of substituted or unsubstituted groups: chemical bond, CH 2 , CHD, CD 2 , C=O, O, NH, SO, SO 2 , P=O, NHCO, NHSO 2 , OCH 2 , OCH 2 CH 2 , CH 2 OCH 2 , NHCH 2 , NMeCH 2 , NHCH 2 CH 2 , NMeCH 2 CH 2 , CH 2 , NHCO, NHCOCH2 ,

[0041]

[0042]

[0043]

[0044]

[0045] where each n is independently 0, 1, 2, 3, 4, 5 or 6; each r and m is independently 0, 1 or 2; each U and W is independently selected from the group consisting of: N or CH;

[0046] p L1 , p L2 , p L3 , p L4 , p L5 and p L6 are independently selected from 0, 1, 2, 3, 4, 5, or 6.

[0047] In some embodiments, V is selected from the group consisting of:

[0048] In some embodiments, X and Y are each independently selected from the group consisting of: N, CH or CR 3 ;

[0049] wherein, R 3 is selected from the group consisting of: halogen, cyano, hydroxy, amino, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 3 alkoxy, halo-C 1 -C 3 alkoxy.

[0050] In some embodiments, B is selected from the group consisting of: NH, O, CO;

[0051] where each U and W is independently selected from the group consisting of: N or CH; each m, p, m', p' is independently 0, 1, 2 or 3.

[0052] In some embodiments, ring A is selected from:

[0053] wherein, W is selected from the group consisting of: CH, N.

[0054] R' is optionally selected from: hydrogen, halogen, cyano, hydroxy, amino, C1 -C 5 alkyl, halo-C 1 -C 5 alkyl, C 1 -C 5 alkoxy, halo-C 1 -C 5 alkoxy, C 3 -C 8 cycloalkyl

[0055] In some embodiments, D, E, F, and G are CH, CR 6 ; wherein, R 6 is selected from the group consisting of: halogen.

[0056] In some embodiments, R 1 is -NHR 7 ;

[0057] wherein, R 7 is -CH 2 R 10 ; R 10 is as defined above.

[0058] In another preferred embodiment, R 10 is selected from the group consisting of: cyano, C 1 -C 5 alkyl, halo-C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 3 -C 10 cycloalkyl, 4-7 membered heterocyclic group, C 6 -C 10 aryl; wherein said aryl is substituted with one or more CR 11 ; R 11 is as defined above.

[0059] In some embodiments,

[0060] when B is selected from the group consisting of: NH, O, CO;

[0061] then the Linker is selected from the group consisting of:

[0062] and when B is then the Linker is selected from the group consisting of: chemical bond, Each n is independently 0, 1, 2, 3, 4, 5 or 6; each r and m are independently 0, 1 or 2; each U and W are independently selected from the group consisting of: N or CH.

[0063] In some embodiments, the compound has the structure shown in formula (II):

[0064]

[0065] Wherein, B is selected from: Wherein, each U and W are independently selected from the group consisting of: N or CH; each m, p, m' and p' are independently 0, 1, 2 or 3;

[0066] R' is optionally selected from: hydrogen, halogen, cyano, hydroxy, amino;

[0067] R 11 、R 6 are each independently selected from: hydrogen, halogen;

[0068] X and Y are independently selected from: CH, N or CR 3 ; wherein, R 3 is optionally selected from: halogen, cyano, hydroxy, amino;

[0069] Z is optionally selected from: CH 2 or CO;

[0070] In some embodiments, the compound is selected from the group consisting of:

[0071]

[0072]

[0073]

[0074]

[0075] In a second aspect of the present invention, there is provided a pharmaceutical composition, which is characterized by comprising

[0076] (1) a compound as described in the first aspect of the present invention as an active ingredient, or a pharmaceutically acceptable salt, its stereoisomer or its prodrug molecule; and

[0077] optionally (2) a pharmaceutically acceptable carrier.

[0078] In a third aspect of the present invention, there is provided an application of a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule thereof, or a pharmaceutical composition as described in the second aspect of the present invention, which is characterized by being used for preparing a CDK12 / 13 protein kinase degrader.

[0079] In a fourth aspect of the present invention, there is provided the use of a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, or a pharmaceutical composition as described in the second aspect of the present invention, characterized in that it is used for preparing a drug for preventing and / or treating a disease mediated by CDK12 / 13 serine / threonine protein kinase.

[0080] In some embodiments, the diseases mediated by CDK12 / 13 serine / threonine protein kinase are selected from the group consisting of: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing's sarcoma, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma, head and neck tumor, colon cancer, rectal cancer, glioma.

[0081] 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 in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0082] Figure 1 (A) WB results of CDK12 protein degradation after treating MDA-MB-231 cells with some compounds at a concentration of 0.1 μM for 15 hours; (B) WB results of CDK13 protein degradation after treating MDA-MB-231 cells with some compounds at a concentration of 0.1 μM for 15 hours; (C) Quantifying the CDK12 / 13 levels by the gray value of the bands in the WB result images (Figure A and Figure B) with the control group DMSO as the reference standard; (D) Compound ZLC5-11 degrades CDK12 and CDK13 proteins in MDA-MB-231 cells in a time-dependent manner.

[0083] Figure 2 WB determination results of intracellular CDK12 and CDK13 protein levels after treating MDA-MB-231 cells with representative compounds ZLC5-6, ZLC5-31, ZLC5-11, ZLC6-37, ZLC6-42 and ZLC5-15 at different concentrations for 15 hours.

[0084] Figure 3 Results of the determination of the inhibitory activity of the compound against triple-negative breast cancer cell MDA-MB-231. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0085] Through extensive and in-depth research, the inventors unexpectedly discovered a class of trans-1,4-cyclohexanediamine compounds. A series of bioactivity tests were conducted on them, and it was found that they have excellent CDK12 / 13 inhibitory performance. Based on this, the present invention was completed.

[0086] In the experimental methods of the following examples of the present invention where specific conditions are not indicated, they are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers. All kinds of commonly used chemical reagents used in the examples are commercially available products.

[0087] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0088] The terms "comprising" and "having" and any variations thereof in the present invention are intended to cover non-exclusive inclusion. For example, a process, method, device, product or equipment comprising a series of steps is not limited to the listed steps or modules, but optionally further includes steps not listed, or optionally further includes other steps inherent to these processes, methods, products or equipment.

[0089] As used in the present invention, "a plurality of" means two or more. "And / or" describes the association relationship of associated objects and indicates that three relationships can exist. For example, A and / or B can mean: A exists alone, A and B exist simultaneously, and B exists alone. The character " / " generally indicates that the associated objects before and after are in an "or" relationship.

[0090] In the compounds of the present invention, when any variable (such as R 10 、R 11 etc.) appears more than once in any component, its definition each time is independent of the definition of each other occurrence. Similarly, combinations of substituents and variables are allowed, provided that such combinations render the compound stable. The line from a substituent into the ring system indicates that the bond referred to can be attached to any ring atom capable of being substituted. If the ring system is polycyclic, it means that such a bond is only attached to any appropriate carbon atom of the adjacent ring. It is to be understood that those of ordinary skill in the art can select the substituents and substitution patterns of the compounds of the present invention to provide compounds that are chemically stable and can be easily synthesized from readily available starting materials by the techniques of the art and the methods presented below. If a substituent itself is substituted by more than one group, it is to be understood that these groups can be on the same carbon atom or different carbon atoms, provided that the structure is stable.

[0091] As used herein, the term "alkyl" is intended to include branched and straight-chain saturated hydrocarbon groups having a specific number of carbon atoms. For example, "C 1 -C8 In “alkyl”, “C” 1 -C 8 The definition of “” includes groups having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms arranged in a straight-chain or branched-chain configuration. The term “cycloalkyl” refers to a monocyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms. For example, “cycloalkyl” includes cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and the like.

[0092] As used herein, the term “alkenyl” includes straight-chain or branched-chain alkenyl groups. For example, C 2 -C 6 Alkenyl refers to a straight-chain or branched-chain alkenyl group having 2-6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or similar groups.

[0093] As used herein, the term “alkynyl” includes straight-chain or branched-chain alkynyl groups. For example, C 2 -C 6 Alkynyl refers to a straight-chain or branched-chain alkynyl group having 2-6 carbon atoms, such as ethynyl, propynyl, butynyl, or similar groups.

[0094] As used herein, the term “cycloalkyl” refers to a cyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms. For example, C 3 -C 10 Alkenyl refers to a cyclic saturated aliphatic hydrocarbon group having 3-10 carbon atoms. It can be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or similar groups. It can also be in a bicyclic form, such as a bridged or spiro form.

[0095] As used herein, the term “heterocyclic group” or “heterocycloalkyl” refers to a saturated or partially saturated cyclic group having a specific number of ring atoms (such as 3-10 ring atoms), and 1-3 of which are heteroatoms selected from N, S, and O. It can be monocyclic, or in a bicyclic or polycyclic form, such as a fused ring, bridged ring, or spiro ring form. Specific examples can be oxetanyl, azetidinyl, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuryl, morpholinyl, and pyrrolidinyl, etc.

[0096] As used herein, the term “alkylamino” refers to an amino group substituted by an alkyl group. For example, “C 1 -C 6 Alkylamino” refers to an amino group substituted by C 1 -C 6 alkyl group, which can be mono-substituted or di-substituted; for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di-tert-butylamino, etc.

[0097] As used herein, the term "alkoxy" refers to a group having an alkyl-oxy structure. For example, "C 1 -C 6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms, including methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, and the like.

[0098] As used herein, the term "haloalkyl" represents an alkyl group in which one or more hydrogen atoms are replaced by halogen, wherein the definition of alkyl is as described above.

[0099] As used herein, the term "haloalkoxy" represents an alkoxy group in which one or more hydrogen atoms are replaced by halogen, wherein the definition of alkoxy is as described above.

[0100] As will be understood by those skilled in the art, "halogen" as used herein means including F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl, and Br. "Halogenated" means being replaced by an atom selected from F, Cl, Br, and I.

[0101] Unless otherwise specified as "substituted or unsubstituted", the groups described in the present invention can be substituted by substituents selected from the following group: halogen, cyano, nitro, hydroxy, amino, C 1 -C 6 alkyl-amino, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 alkoxy, halo C 1 -C 6 alkyl, halo C 2 -C 6 alkenyl, halo C 2 -C 6 alkynyl, halo C 1 -C 6 alkoxy, allyl, benzyl, C 6 -C 12 aryl, C 1 -C 6 alkoxy-C 1 -C 6 alkyl, C 1 -C 6 alkoxy-carbonyl, phenoxycarbonyl, C 2 -C 6 alkynyl-carbonyl, C 2 -C 6Alkenyl-carbonyl, C 3 -C 6 Cycloalkyl-carbonyl, C 1 -C 6 Alkyl-sulfonyl, etc.

[0102] The present invention includes the free form of the compounds of formula (I) (wherein formula (I) includes formula (II)), as well as their pharmaceutically acceptable salts, their stereoisomers, and their prodrug molecules. The term "free form" refers to the compound in non-salt form. The pharmaceutically acceptable salts included not only include the exemplary salts of the specific compounds described herein, but also include the typical pharmaceutically acceptable salts of all free forms of the compounds of formula (I) or formula (II). The free form of the specific salts of the compounds can be separated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a dilute aqueous solution of a suitable base such as dilute aqueous NaOH, dilute aqueous potassium carbonate, dilute aqueous ammonia, and dilute aqueous sodium bicarbonate. The free form is somewhat different from its respective salt form in certain physical properties such as solubility in polar solvents, but for the purposes of the invention, such acid salts and base salts are equivalent to their respective free forms in other pharmaceutical aspects.

[0103] The pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of the present invention containing a basic moiety or an acidic moiety by conventional chemical methods. Generally, salts of basic compounds are prepared by ion exchange chromatography or by reacting the free base with a stoichiometric or excess amount of the desired salt form of an inorganic or organic acid in a suitable solvent or a combination of solvents. Similarly, salts of acidic compounds are formed by reacting with a suitable inorganic or organic base.

[0104] Accordingly, the pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts of the compounds of the present invention formed by reacting a basic compound of the present invention with an inorganic or organic acid. For example, the conventional non-toxic salts include salts obtained from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc., and also include salts obtained from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxy-benzoic acid, fumaric acid, benzenesulfonic acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, 2-hydroxyethanesulfonic acid, trifluoroacetic acid, etc.

[0105] If the compound of the present invention is acidic, suitable "pharmaceutically acceptable salts" refer to salts prepared with pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. Ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases, said bases including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0106] Berg et al., "Pharmaceutical Salts" J. Pharm. Sci. ’1977:66:1-19 describes in more detail the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts.

[0107] Unless otherwise specified, the structural formulas described in the present invention are intended to include all isomeric forms (such as enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, the R and S configurations containing asymmetric centers, the (Z) and (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers or geometric isomers (or conformational isomers) are within the scope of the present invention.

[0108] As used herein, the term "tautomer" means that structural isomers with different energies can cross a low energy barrier and thus interconvert. For example, proton tautomers (i.e., prototropy) include interconversion through proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion through some reorganization of bonding electrons.

[0109] As used herein, the term "solvate" refers to a complex formed by the coordination of a compound of the present invention with solvent molecules in a specific ratio.

[0110] As used herein, the term "hydrate" refers to a complex formed by the coordination of a compound of the present invention with water.

[0111] The compounds of the present invention may also be in the form of prodrugs. As used herein, the term "prodrug" refers to a compound that produces an active compound upon metabolism (e.g., in vivo or in vitro). In some embodiments, the prodrug may be inactive or have lower activity than the free drug, but may provide advantageous handling, administration, or metabolic characteristics. Exemplary prodrug moieties of the present invention may be linked to the free drug through the hydroxyl, amino, phosphate, or phosphorothioate backbone of a nucleotide and may include esters, carbamates, carbonyls, thioesters, amides, isocyanates, ureas, thioureas, or other physiologically acceptable metabolically labile moieties. In some embodiments, the prodrug is activated by enzymatic hydrolysis.

[0112] The present disclosure also includes isotopically labeled compounds that are identical to the compounds of formula (I) (including the compounds of formula (II)), but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as but not limited to 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 31 P, 35 S, 18 F, and 36 Cl. Replacement with a heavier isotope (e.g., deuterium, i.e., 2 H) may provide certain therapeutic advantages, such as higher metabolic stability, e.g., a longer in vivo half-life or a lower dose requirement, and may thus be preferred in certain cases. The compounds described may incorporate positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies to determine the distribution of receptors. Suitable positron-emitting isotopes that may be incorporated into the compounds of formula (I) or (II) are 1 C, 13 N, 15 O, and 18 F. Isotopically labeled compounds of formula (I) or (II) can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described herein, using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents.

[0113] The compounds disclosed herein may exist in solvated and unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc., and the present invention is intended to cover solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in crystalline form.

[0114] Drug Compositions and Administration Methods

[0115] Since the compounds of the present invention are CDK12 / 13 protein degraders, the compounds and their pharmaceutically acceptable salts and other compound forms disclosed herein may be included in pharmaceutical compositions useful for treating, preventing, and alleviating diseases associated with CDK12 / 13 activity.

[0116] The pharmaceutical compositions of the present invention comprise an effective amount, such as a safe and effective amount, of a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier. "Effective amount" means an amount sufficient to elicit the desired biological response (e.g., treating a disorder). "Safe and effective amount" means that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition comprises 1 to 3000 mg (active dose range of 3 to 30 mg / kg) of the disclosed compound / dose, and more preferably comprises 10 to 2000 mg of the compound of the present invention / dose. Preferably, "one dose" is a capsule or a tablet.

[0117] "Pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers or gelling substances suitable for human use, which must have sufficient purity and sufficiently low toxicity. "Compatibility" herein means that the components of the composition can be mixed with the compounds disclosed herein and with each other without significantly reducing the efficacy of the compounds. Examples of parts of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, and cellulose acetate), gelatin, talc, solid lubricants (such as stearic acid and magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil), polyols (propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers ( etc.), wetting agents (sodium lauryl sulfate, etc.), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, and pyrogen-free water.

[0118] There is no particular limitation on the administration method of the compounds or pharmaceutical compositions of the present invention, and representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0119] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or calcium phosphate dibasic, or is admixed with the following components: (a) fillers or fillers, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxypropylmethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, some complex silicates, and sodium carbonate; (e) retardant solvents, such as paraffin wax; (f) absorption promoters, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also include buffering agents.

[0120] Solid dosage forms, such as tablets, dragees, capsules, pills, and granules, may be prepared with coatings and shell materials, such as enteric coatings and other materials well known in the art. It may contain opacifying agents, and active compounds or compounds that are released in a delayed manner in a part of the digestive tract in such compositions. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0121] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 - butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof.

[0122] In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and perfumes.

[0123] In addition to the active compound, suspensions may also contain suspending agents, such as ethoxylated isooctadecanol, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum monostearate, and agar, or mixtures thereof.

[0124] Compositions for parenteral injection may contain physiologically acceptable sterile aqueous or non - aqueous solutions, dispersions, suspensions, or emulsions, as well as 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.

[0125] The topical dosage forms of the compounds of the present invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed with a physiologically acceptable carrier and any preservatives, buffers or propellants (if required) under sterile conditions.

[0126] The compounds of the present disclosure can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0127] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applicable to mammals (such as humans) in need of treatment, wherein the dose is a dose considered pharmaceutically effective when administered. The daily dose for a 60 kg human is generally 1 - 2000 mg, preferably 6 - 600 mg. Of course, the specific dose should also be determined by comprehensively considering factors such as the administration route and the health status of the patient, which are within the capabilities of a skilled physician.

[0128] Uses and treatment methods

[0129] As described above, the compounds of the present invention are CDK12 / 13 protein degraders, and thus the compounds or compositions containing the compounds can be used to treat, prevent and alleviate diseases related to CDK12 / 13 activity or abnormal expression. In some embodiments, the use of the compounds of the present invention in the preparation of a medicament for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinase is disclosed herein. In some embodiments, the compounds of the present invention for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinase are disclosed herein. In some embodiments, a method for treating diseases mediated by CDK12 / 13 serine / threonine protein kinase in a subject in need thereof is disclosed herein, comprising administering to the subject an effective amount of the compound of the present invention. In some embodiments, diseases mediated by CDK12 / 13 serine / threonine protein kinase include: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing's sarcoma, lung adenocarcinoma, squamous cell lung cancer, pancreatic cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma, head and neck tumors, colon cancer, rectal cancer, glioma.

[0130] When used for the disclosed uses and methods, the disclosed compounds and compositions can be used in combination with other known therapies. As used herein, "combination" administration refers to the delivery of two (or more) different treatments to a subject during the course of a disorder in the subject, e.g., two or more treatments are carried out after the subject has been diagnosed with a disorder and before the disorder has been cured or eliminated, or treatment has stopped for other reasons. In some embodiments, when the delivery of the second treatment begins, the delivery of one treatment is still ongoing, so there is an overlap in administration. This is sometimes referred to herein as "simultaneous" or "concurrent delivery". In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective due to the combination administration. For example, the second treatment is more effective, e.g., the same effect can be seen with less of the second treatment compared to performing the second treatment without the first treatment, or the second treatment can reduce symptoms to a greater extent, or for a similar situation observed with the first treatment. In some embodiments, the delivery is such that the reduction in symptoms or other parameters associated with the disorder is greater than that observed with one treatment delivered without the other treatment. The effects of the two treatments can be partially additive, fully additive, or more than additive. The delivery can be such that the effect of the first treatment delivered is still detectable when the second treatment is delivered.

[0131] The compounds or compositions disclosed herein and at least one additional therapeutic agent can be administered simultaneously, in the same or separate compositions, or sequentially. For sequential administration, the compounds described herein can be administered first, followed by the additional agent, or the order of administration can be reversed.

[0132] In some embodiments, the compounds described herein are administered in combination with other treatment modalities, including surgery, radiation, transplantation (e.g., stem cell transplantation, bone marrow transplantation), chemotherapy, immunotherapy, cryotherapy, and / or thermotherapy. Such combination therapies can permit lower doses of the administered agent and / or other agents, thereby avoiding possible toxicities or complications associated with the various therapies.

[0133] In some embodiments, the compounds described herein are administered with at least one additional therapeutic agent, such as a chemotherapeutic agent. In certain embodiments, the compounds described herein are administered in combination with one or more additional chemotherapeutic agents. The chemotherapeutic agent can be a chemotherapeutic agent identified in the "A to Z List of Cancer Drugs" published by the National Cancer Institute of the United States.

[0134] The main advantages of the present invention are:

[0135] 1. The degrader of cyclin-dependent protein kinase 12 / 13 (CDK12 / 13) provided by the present invention can effectively degrade CDK12 and CDK13 protein kinases, and can be used to prepare drugs for preventing or treating diseases mediated by CDK12 and / or CDK13 protein kinases, such as prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing's sarcoma, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma, head and neck tumors, colon cancer, rectal cancer, glioma, etc.

[0136] 2. The degrader of cyclin-dependent protein kinase 12 / 13 (CDK12 / 13) provided by the present invention has strong degradation activity.

[0137] 3. The degrader of cyclin-dependent protein kinase 12 / 13 (CDK12 / 13) provided by the present invention has high selectivity for protein kinase degradation.

[0138] The following further describes the present invention in detail with specific examples.

[0139] Example 1: Compound zlc-4-77

[0140]

[0141] Step 1: Preparation of intermediate 3-86

[0142]

[0143] In a two-necked round-bottom flask, to a system of 4-(4-bromophenyl)piperazine-1-carboxylic acid tert-butyl ester (1.02 g, 3 mmol) in anhydrous DMSO (15 mL), trans-cyclohexane-1,4-diamine 2 (1.2 g, 10.5 mmol), potassium phosphate (1.3 g, 6 mmol), CuI (57 mg, 0.3 mmol), and D-proline (35 mg, 0.3 mmol) were added in sequence. After completion, the gas was replaced with argon three times, and the temperature was raised to 100 °C. After monitoring the reaction completion by TLC, it was transferred to room temperature, filtered through diatomaceous earth, washed three times with a DCM / MeOH (10:1) system, the filtrate was concentrated under reduced pressure, loaded onto the column by wet method, and purified by silica gel normal-phase chromatography column to obtain the target compound 3-86 (off-white solid, 480 mg, yield 43%). 1 H NMR(400MHz, DMSO-d 6)δ 6.74 (d, J = 8.8 Hz, 2H), 6.49 (d, J = 8.9 Hz, 2H), 4.88 (d, J = 8.2 Hz, 1H), 3.42 (t, J = 5.1 Hz, 4H), 3.03 (s, 1H), 2.83 (t, J = 5.1 Hz, 4H), 2.76 (s, 1H), 1.95 (d, J = 12.8 Hz, 2H), 1.85 (d, J = 12.4 Hz, 2H), 1.41 (s, 9H), 1.26 (q, J = 10.9 Hz, 2H), 1.11 (q, J = 11.6 Hz, 2H). HRMS(ESI) for C 21 H 34 N 4 O 2 [M + H] + , calcd: 375.2755, found: 375.2739.

[0144] Step 2: Preparation of Intermediate 4-39

[0145]

[0146] In a 100 mL round-bottom flask, to the DMF (7.5 mL) system of 3-86 (1.38 g, 3.7 mmol), 5-cyano-2-fluoropyridine 3 (451 mg, 3.7 mmol) and Cs 2 CO 3 (1.45 g, 4.44 mol) were added successively. After completion, the reaction was carried out at room temperature overnight. After monitoring the completion of the reaction by TLC, the mixture was filtered and the filtrate was concentrated under reduced pressure. Purification by silica gel column chromatography in normal phase chromatography gave Intermediate 4-39 (white solid, 1.4 g, 80%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.37 (s, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.51 (d, J = 1 Hz, 1H), 6.75 (d, J = 6.9 Hz, 2H), 6.52 (d, J = 8.3 Hz, 3H), 4.92 (s, 1H), 3.75 (s, 1H), 3.42 (s, 4H), 3.12 (s, 1H), 2.83 (s, 4H), 1.98 (s, 4H), 1.41 (s, 9H), 1.36 - 1.28 (m, 2H), 1.25 - 1.17 (m, 2H).

[0147] Step 3: Preparation of Intermediate 4-44

[0148]

[0149] In a 25 mL round-bottom flask, benzyl isocyanate 4 (1.2 g, 9 mmol) and DIPEA (1.6 mL) were successively added to a DMF (3 mL) system of 4-39 (1.438 g, 3 mmol). After completion, the temperature was raised to 95 °C. After 6 h, the reaction was monitored by TLC to be complete. It was transferred to room temperature, concentrated under reduced pressure, and purified by silica gel normal-phase chromatography to obtain the target intermediate 4-44 (white solid, 1.3 g, yield 72%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.29 (s, 1H), 7.60 (d, J = 9.0 Hz, 1H), 7.46 (d, J = 6.6 Hz, 1H), 7.28 - 7.24 (m, 2H), 7.18 - 7.14 (m, 3H), 7.03 - 6.98 (m, 4H), 6.46 (d, J = 8.7 Hz, 1H), 5.58 - 5.56 (m, 1H), 4.26 (t, J = 11.9 Hz, 1H), 4.14 (d, J = 5.4 Hz, 2H), 3.45 (s, 5H), 3.16 (s, 4H), 1.90 (d, J = 11.1 Hz, 2H), 1.76 (d, J = 11.6 Hz, 2H), 1.42 (d, J = 1.6 Hz, 9H), 1.27 (m, 2H), 1.14 - 1.04 (m, 2H).

[0150] Step 4: Preparation of intermediate 4-46

[0151]

[0152] In a 25 mL round-bottom flask, CF 2 Cl 2 COOH (1 mL) was added dropwise to a CH 3 Cl 2 (2 mL) system of 4-44 (1.83 g, 3 mmol). Stir at room temperature overnight. After the reaction was complete, it was concentrated under reduced pressure and extracted with a saturated sodium bicarbonate-CH 2 Cl 1 / MeOH (10∶1) system. It was dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain the target intermediate 4-46 (white solid, 1.43 g, 94%). 6)δ8.29(s, 1H), 7.60(d, J = 9.2Hz, 1H), 7.48(s, 1H), 7.28 - 7.25(m, 2H), 7.18 - 7.15(m, 3H), 7.00 - 6.95(m, 4H), 6.47(d, J = 8.9Hz, 1H), 5.56(s, 1H), 4.26(m, 1H), 4.14(d, J = 5.9Hz, 2H), 3.48(s, 1H), 3.37 - 3.34(m, 1H), 3.11 - 3.09(m, 4H), 2.85 - 2.83(m, 4H), 1.90(d, J = 11.4Hz, 2H), 1.76(d, J = 10.7Hz, 2H), 1.33 - 1.27(m, 2H), 1.13 - 1.06(m, 2H).

[0153] Step 5: Preparation of Intermediate 4 - 65

[0154]

[0155] In a 25 mL round - bottom flask, to the DMF (2 mL) system of 4 - 46 (153 mg, 0.3 mmol), potassium carbonate (83 mg, 0.6 mmol) and 1 - Boc - 4 - bromomethylpiperidine (167 mg, 0.6 mmol) were added successively. After completion, the temperature was raised to 80 °C and stirred overnight. After the reaction was completed, the reaction solution was concentrated under reduced pressure and purified by normal - phase silica gel column chromatography to obtain the target intermediate 4 - 65 (white solid, 110 mg, 52%). 1 H NMR(500MHz, DMSO - d 6 )δ8.29(d, J = 2.1Hz, 1H), 7.60(d, J = 8.7Hz, 1H), 7.49(d, J = 6.6Hz, 1H), 7.28 - 7.26(m, 2H), 7.18 - 7.14(m, 3H), 7.03 - 6.96(m, 4H), 6.46(d, J = 8.8Hz, 1H), 5.59(t, J = 5.6Hz, 1H), 4.28 - 4.23(s, 1H), 4.14 - 4.13(d, J = 6.0Hz, 2H), 3.92(s, 2H), 3.50(s, 1H), 3.17(s, 4H), 2.47(s, 4H), 2.17(d, J = 6.6Hz, 2H), 1.90(d, J = 9.7Hz, 2H), 1.77 - 1.68(m, 5H), 1.39(s, 9H), 1.33 - 1.26(m, 2H), 1.13 - 1.08(m, 2H), 0.98 - 0.94(m, 2H), 0.84 - 0.81(m, 2H). MS(ESI) for C 41 H 55 N 8O 3 [M+H] + ,calcd:707.4,found:707.5.

[0156] Step 6: Preparation of compound zlc-4-77

[0157]

[0158] In a 25 mL round-bottom flask, to the system of 4-65 (79 mg, 0.112 mmol) in CH 2 Cl 2 (1 mL) was added dropwise CF 3 COOH (0.3 mL). The mixture was stirred at room temperature overnight. After the reaction was completed, the solvent was concentrated under reduced pressure. Then, DMF (2 mL), 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione (31 mg, 0.112 mmol), and DIPEA (0.023 mL, 0.135 mmol) were added successively. The temperature was raised to 105 °C. After monitoring the completion of the reaction, the mixture was transferred to room temperature and concentrated under reduced pressure. The target product zlc-4-77 (yellow solid, 30 mg, 32%) was obtained by normal-phase silica gel chromatography purification. 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 8.29 (d, J = 2.1 Hz, 1H), 7.65 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.47 (s, 1H), 7.31 (s, 1H), 7.28 - 7.25 (m, 2H), 7.23 (d, J = 8.6 Hz, 1H), 7.18 - 7.15 (m, 3H), 7.01 - 6.97 (m, 4H), 6.51 - 6.44 (m, 1H), 5.57 (t, J = 5.2 Hz, 1H), 5.06 (dd, J = 12.8, 5.4 Hz, 1H), 4.26 (t, J = 11.8 Hz, 1H), 4.15 (d, J = 5.7 Hz, 2H), 4.05 (d, J = 12.7 Hz, 2H), 3.48 (s, 1H), 3.19 (s, 4H), 2.97 (t, J = 12.0 Hz, 2H), 2.93 - 2.84 (m, 1H), 2.63 - 2.53 (m, 2H), 2.52 - 2.50 (m, 4H), 2.20 (d, J = 6.1 Hz, 2H), 2.04 - 1.99 (m, 1H), 1.90 (d, J = 10.3 Hz, 3H), 1.82 (d, J = 12.5 Hz, 2H), 1.76 (d, J = 10.6 Hz, 2H), 1.35 - 1.28 (m, 2H), 1.17 - 1.05 (m, 4H). 1313C NMR (151 MHz, DMSO) δ 172.81, 170.12, 167.64, 166.97, 159.25, 156.80, 155.01, 153.10, 150.15, 141.33, 134.05, 131.50, 128.15, 128.02, 126.69, 126.23, 125.01, 119.12, 117.58, 117.35, 115.34, 107.74, 93.99, 63.71, 53.21, 52.91, 48.73, 47.70, 47.27, 43.47, 32.49, 31.31, 30.98, 30.20, 29.64, 22.20, 14.09. HRMS (ESI) for C 49 H 55 N 10 O 5 [M + H] + , calcd: 863.4351, found: 863.4360.

[0159] Example 2: Preparation of Compound zlc - 4 - 79

[0160]

[0161] Step 1: Preparation of Intermediate 4 - 66

[0162] In a 25 mL round - bottom flask, to the DMF (2 mL) solution of 4 - 46 (153 mg, 0.3 mmol), HATU (137 mg, 0.36 mmol), DIPEA (0.1 mL, 0.6 mmol) and 1 - N - Boc - azetidine - 3 - carboxylic acid (67 mg, 0.33 mmol) were added successively. After completion, the reaction was stirred at room temperature. After monitoring the completion of the reaction, water was added to quench the reaction, and a solid was precipitated. The solid was filtered, and the filter cake was washed (CH 2 Cl 2 / MeOH = 10:1), dried over anhydrous sodium sulfate, and purified by normal - phase silica gel chromatography to obtain the target intermediate 4 - 66 (white solid, 120 mg, 59%). 1 1H NMR (500 MHz, DMSO - d 6)δ8.29(d, J = 2.1Hz, 1H), 7.60(d, J = 9.2Hz, 1H), 7.48(s, 1H), 7.28 - 7.25(m, 2H), 7.18 - 7.14(m, 3H), 7.03 - 6.99(m, 4H), 6.46(d, J = 8.9Hz, 2H), 5.61 - 5.59(m, 1H), 4.28 - 4.23(m, 1H), 4.14(d, J = 5.8Hz, 2H), 3.72 - 3.68(m, 1H), 3.63 - 3.60(m, 4H), 3.40(s, 2H), 3.19 - 3.17(m, 3H), 3.16 - 3.12(m, 4H), 1.90(d, J = 10.9Hz, 2H), 1.76(d, J = 12.3Hz, 2H), 1.37(s, 9H), 1.32 - 1.27(m, 2H), 1.13 - 1.06(m, 2H). MS(ESI) for C 39 H 49 N 8 O 4 [M + H] + , calcd: 693.4, found: 692.8.

[0163] Step 2: Preparation of compound zlc - 4 - 79

[0164]

[0165] The synthesis method is the same as that in step f of Example 1.

[0166] 1 H NMR(600MHz, DMSO - d 6)δ 11.07 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.51 - 7.44 (m, 1H), 7.28 - 7.26 (m, 2H), 7.19 - 7.15 (m, 3H), 7.05 - 7.01 (m, 4H), 6.84 (d, J = 1.8 Hz, 1H), 6.70 (dd, J = 8.4, 2.0 Hz, 1H), 6.47 (d, J = 8.9 Hz, 1H), 5.58 (t, J = 5.5 Hz, 1H), 5.06 (dd, J = 12.8, 5.4 Hz, 1H), 4.28 - 4.25 (m, 3H), 4.18 - 4.15 (m, 4H), 4.00 - 3.97 (m, 1H), 3.65 (s, 2H), 3.49 (s, 3H), 3.22 (m, 4H), 2.91 - 2.84 (m, 1H), 2.59 - 2.53 (m, 2H), 2.03 - 2.00 (m, 1H), 1.91 (d, J = 10.3 Hz, 2H), 1.77 (d, J = 10.6 Hz, 2H), 1.31 (m, 2H), 1.10 (m, 2H). 13 C NMR (151 MHz, DMSO) δ 172.80, 170.10, 169.35, 167.43, 167.15, 159.24, 156.77, 154.98, 153.10, 149.92, 141.30, 133.80, 131.60, 128.76, 128.03, 126.71, 126.25, 124.83, 119.12, 117.24, 115.97, 114.30, 104.56, 94.00, 53.63, 52.92, 48.72, 48.07, 47.75, 44.42, 43.48, 41.30, 31.30, 31.21, 30.98, 30.20, 22.19, 14.09. HRMS (ESI) for C 47 H 49 N 10 O 6 [M + H] + , calcd: 849.3831, found: 849.3839.

[0167] Example 3: Preparation of Compound zlc - 4 - 91

[0168]

[0169] Step 1: Preparation of Intermediate 4 - 64

[0170]

[0171] The compound p-bromoiodobenzene (5.0 g, 17.7 mmol), N-BOC-trans-1,4-cyclohexanediamine (3.2 g, 14.7 mmol), Pd 2 (dba) 3 (1.37 g, 1.5 mmol), Xantphos (1.7 g, 2.94 mmol) and tert-ButONa (2.8 g, 29.4 mmol) were mixed and dissolved in 150 mL of toluene. After purging with argon three times, the reaction solution was heated to 100 °C and reacted overnight. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, and the reaction solution was concentrated by rotary evaporation under reduced pressure. The target compound 4-64 (3.9 g of yellow solid, yield 72%) was obtained by column chromatography purification. 1 1H NMR (500 MHz, DMSO-d 6 ) δ 7.16 - 7.13 (m, 2H), 6.79 (d, J = 7.9 Hz, 1H), 6.49 (dd, J = 8.9, 2.0 Hz, 2H), 5.63 (d, J = 8.0 Hz, 1H), 3.21 - 3.19 (m, 1H), 3.06 - 3.03 (m, lH), 1.93 (d, J = 12.6 Hz, 2H), 1.77 (d, J = 12.5 Hz, 2H), 1.28 - 1.21 (m, 1H), 1.16 - 1.11 (m, 2H).

[0172] Step 2: Preparation of Intermediate 4-68

[0173]

[0174] Intermediate 4-64 (3.9 g, 10.6 mmol) was dissolved in 4 mL of DMF, and benzyl isocyanate (4.2 g, 31.7 mmol), DIPEA (1.59 g, 12.3 mmol) were added, and the reaction was carried out at 95 °C for 5 h. The solvent was evaporated to dryness, and the target compound 4-68 (3.7 g of yellow solid, yield 69%) was obtained by column chromatography. 1 1H NMR (500 MHz, DMSO-d 6 ) δ 7.64 - 7.62 (m, 2H), 7.27 - 7.25 (m, 2H), 7.19 - 7.15 (m, 3H), 7.12 - 7.10 (m, 2H), 6.69 (d, J = 7.9 Hz, 1H), 6.01 (t, J = 6.1 Hz, 1H), 4.24 - 4.16 (m, 1H), 4.13 (d, J = 6.1 Hz, 2H), 2.94 (m, 1H), 1.72 (d, J = 11.7 Hz, 4H), 1.34 (s, 9H), 1.24 - 1.21 (m, 2H), 1.03 - 0.96 (d, J = 12.1 Hz, 2H).

[0175] Step 3: Preparation of Intermediate 4-69

[0176]

[0177] Dissolve Intermediate 4-68 (3.7 g, 7.3 mmol) in 5 mL of DCM, and add 2.5 mL of trifluoroacetic acid (TFA). After heating to reflux at 55 °C for 6 h, the solvent was evaporated under reduced pressure, and the target compound 4-69 (2.5 g of yellow solid, yield 70%) was obtained by column chromatography. 1 HNMR (500 MHz, DMSO-d 6 ) δ 7.65 - 7.63 (m, 2H), 7.28 - 7.25 (m, 2H), 7.19 - 7.11 (m, 5H), 6.04 (m, 1H), 4.23 - 4.18 (m, 1H), 4.12 (d, J = 5.3 Hz, 2H), 2.79 (m, 1H), 1.89 (d, J = 12.8 Hz, 2H), 1.79 (d, J = 12.2 Hz, 2H), 1.38 (m, 2H), 1.09 - 1.01 (m, 2H).

[0178] Step 4: Preparation of Intermediate 4-87

[0179]

[0180] Dissolve Compound 4-69 (2.5 g, 5.1 mmol) in 15 mL of DMF, add 5-cyano-2-fluoropyridine (744 mg, 6.1 mol), Cs 2 CO 3 (2.0 g, 6.1 mol). The mixture was stirred at room temperature for 15 min and then heated to 60 °C for 40 min. After the reaction was completed, the solvent was evaporated under reduced pressure, and the target compound 4-87 (1.9 g of white solid, yield 70%) was obtained by column chromatography purification. 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.30 (d, J = 2.2 Hz, 1H), 7.65 - 7.60 (m, 3H), 7.48 (d, J = 6.5 Hz, 1H), 7.29 - 7.26 (m, 2H), 7.19 - 7.13 (m, 5H), 6.47 (d, J = 8.9 Hz, 1H), 6.03 (t, J = 5.9 Hz, 1H), 4.30 - 4.23 (m, 1H), 4.14 (d, J = 6.0 Hz, 2H), 3.51 (s, 1H), 1.91 (d, J = 11.0 Hz, 1H), 1.78 (d, J = 11.4 Hz, 2H), 1.31 (m, 2H), 1.10 (m, 2H).

[0181] Step 5: Preparation of Intermediate 4-88

[0182]

[0183] Into a 25 mL Schlenk flask, add successively compound 4-87 (1.51 g, 3 mmol), tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate (986 mg, 3.6 mmol), Pd 2 (dba) 3 (274 mg, 0.3 mmol), Xantphos (347 mg, 0.6 mmol) and tert-ButONa (576 g, 6 mmol). Replace the atmosphere with argon three times, add toluene (6 mL), heat up to 110 °C and react overnight. After completion, filter through diatomaceous earth, concentrate under reduced pressure, and purify by column chromatography to obtain the target compound 4-88 (yellow solid, 1.27 g, 61%). 1 1H NMR (500 MHz, DMSO-d 6 ) δ 8.29 (d, J = 1.7 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.48 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.14 (m, 3H), 7.00 - 6.95 (m, 5H), 6.46 (d, J = 9.0 Hz, 1H), 5.59 (s, 1H), 4.28 - 4.23 (m, 1H), 4.14 (d, J = 5.9 Hz, 2H), 3.95 (s, 2H), 3.16 (s, 4H), 2.73 (s, 1H), 2.61 (s, 4H), 2.40 - 2.37 (m, 1H), 1.90 (d, J = 10.8 Hz, 2H), 1.77 (m, 4H), 1.39 (s, 9H), 1.33 - 1.23 (m, 6H), 1.13 - 1.05 (m, 2H).

[0184] Step 6: Preparation of Compound zlc-4-91

[0185]

[0186] The synthesis method is the same as that of Step f in Example 1.

[0187] 1 1H NMR (600 MHz, DMSO-d 6)δ11.11(s, 1H), 8.29(d, J = 2.2Hz, 1H), 7.71(d, J = 11.4Hz, 1H), 7.59(d, J = 8.4Hz, 1H), 7.45(m, 2H), 7.28 - 7.25(m, 2H), 7.18 - 7.15(m, 3H), 7.01 - 6.96(m, 4H), 6.46(d, J = 8.9Hz, 1H), 5.57(m, 1H), 5.11(dd, J = 12.9, 5.4Hz, 2H), 4.28 - 4.26(m, 1H), 4.15(d, J = 5.7Hz, 2H), 3.67(d, J = 11.5Hz, 2H), 3.49(s, 1H), 3.19(s, 4H), 2.93 - 2.86(m, 3H), 2.66(s, 4H), 2.61 - 2.58(m, 1H), 2.54 - 2.52(d, J = 13.2Hz, 1H), 2.49 - 2.44(d, J = 15.7Hz, 1H), 2.04 - 2.02(m, 1H), 1.93 - 1.90(m, 4H), 1.76(d, J = 10.4Hz, 2H), 1.62 - 1.57(m, 2H), 1.33 - 1.27(m, 2H), 1.13 - 1.07(d, 2H). 13 C NMR(151MHz, DMSO)δ172.76, 169.91, 166.69, 166.21, 159.25, 158.09, 156.81, 156.41, 153.09, 150.15, 145.47, 141.34, 131.50, 128.77, 128.14, 128.01, 126.69, 126.22, 122.98, 119.11, 115.29, 113.72, 112.00, 111.83, 93.99, 59.76, 52.92, 49.33, 49.05, 48.77, 48.02, 43.48, 31.31, 30.96, 30.20, 27.88, 22.09, 14.09. HRMS(ESI) for C 48 H 52 N 10 O 5 [M + H] + , calcd: 867.4101, found: 867.4104.

[0188] Example 4: Preparation of Compound zlc - 4 - 93

[0189]

[0190] The synthesis method refers to Example 1.

[0191] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.47 (s, 1H), 7.33 (s, 1H), 7.27 - 7.25 (m, 3H), 7.18 - 7.15 (m, 3H), 7.00 - 6.95 (m, 4H), 6.46 (d, J = 8.9 Hz, 1H), 5.58 - 5.56 (m, 1H), 5.08 - 5.05 (dd, J = 12.8, 5.5 Hz, 1H), 4.28 - 4.23 (m, 1H), 4.14 (d, J = 5.8 Hz, 2H), 4.09 (d, J = 12.5 Hz, 2H), 3.48 (s, 1H), 3.17 (s, 4H), 3.01 - 2.97 (m, 2H), 2.91 - 2.85 (m, 1H), 2.63 (s, 4H), 2.60 - 2.50 (m, 4H), 2.03 - 2.00 (m, 1H), 1.90 (d, J = 10.3 Hz, 4H), 1.76 (d, J = 10.5 Hz, 2H), 1.51 - 1.46 (m, 2H), 1.33 - 1.27 (m, 2H), 1.12 - 1.06 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 172.81, 170.11, 167.61, 166.96, 159.24, 156.80, 154.75, 153.09, 150.14, 141.34, 134.04, 131.49, 128.14, 128.01, 126.69, 126.22, 125.01, 119.12, 117.70, 115.30, 107.82, 93.98, 59.76, 52.90, 48.74, 48.00, 46.61, 43.47, 31.30, 30.98, 30.19, 27.21, 22.19, 14.09. HRMS (ESI) for C 48 H 53 N 10 O 5 [M + H] + , calcd: 849.4195, found: 849.4199.

[0192] Example 5: Preparation of Compound zlc - 5 - 6

[0193]

[0194] The synthesis method refers to Example 1.

[0195] 1 ¹H NMR (500 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 10.2 Hz, 1H), 7.47 (d, J = 7.0 Hz, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.02 - 6.97 (m, 4H), 6.78 (d, J = 1.5 Hz, 1H), 6.65 (dd, J = 8.4, 1.7 Hz, 1H), 6.47 (d, J = 8.8 Hz, 1H), 5.58 (m, 1H), 5.07 - 5.03 (m, 1H), 4.29 - 4.24 (m, 1H), 4.17 - 4.14 (m, 4H), 3.73 - 3.70 (m, 2H), 3.50 (s, 1H), 3.50 - 3.46 (m, 1H), 3.19 (s, 4H), 3.05 (s, 1H), 2.90 - 2.84 (m, 1H), 2.66 (d, J = 5.4 Hz, 2H), 2.60 - 2.54 (m, 6H), 2.02 - 2.00 (m, 1H), 1.91 (d, J = 9.7 Hz, 2H), 1.77 (d, J = 10.2 Hz, 2H), 1.33 - 1.26 (m, 2H), 1.13 - 1.06 (m, 2H). 13 ¹³C NMR (151 MHz, DMSO) δ 172.80, 170.11, 167.49, 167.18, 159.24, 156.80, 155.19, 153.10, 150.12, 141.34, 133.82, 131.50, 128.22, 128.02, 126.69, 126.23, 124.80, 119.12, 116.68, 115.42, 114.06, 104.35, 93.98, 61.76, 55.75, 52.85, 48.70, 47.63, 43.46, 31.30, 30.98, 30.20, 27.00, 22.21. HRMS (ESI) for C 47 H 51 N 10 O 5 [M + H] + , calcd: 835.4038, found: 835.4043.

[0196] Example 6: Preparation of Compound zlc - 5 - 11

[0197]

[0198] The synthesis method refers to Example 1.

[0199] 1 H NMR (500 MHz, DMSO-d 6 ) δ 11.09 (s, 1H), 8.31 - 8.28 (m, 1H), 7.68 (d, J = 8.7 Hz, 1H), 7.60 (d, J = 8.8 Hz, 1H), 7.49 (s, 1H), 7.34 (s, 1H), 7.28 - 7.25 (m, 3H), 7.18 - 7.15 (m, 3H), 6.99 (m, 4H), 6.46 (d, J = 8.8 Hz, 1H), 5.59 - 5.57 (m, 1H), 5.07 (dd, J = 12.8, 5.4 Hz, 1H), 4.28 - 4.14 (m, 1H), 4.14 (d, J = 5.6 Hz, 2H), 3.79 (d, J = 9.4 Hz, 2H), 3.44 (m, 4H), 2.92 - 2.85 (m, 1H), 2.75 - 2.70 (m, 2H), 2.65 (m, 4H), 2.60 - 2.56 (m, 2H), 2.41 - 2.36 (m, 1H), 2.03 - 2.00 (m, 1H), 1.91 (s, 4H), 1.76 (d, J = 11.0 Hz, 2H), 1.53 - 1.51 (m, 2H), 1.33 - 1.25 (m, 2H), 1.13 - 1.06 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 172.87, 170.14, 167.60, 167.02, 159.26, 156.85, 153.14, 150.15, 141.36, 133.87, 131.53, 128.06, 127.91, 126.71, 126.27, 124.94, 119.17, 115.82, 94.00, 59.81, 52.91, 48.79, 48.42, 47.47, 43.49, 31.34, 31.01, 30.24, 22.21, 14.13. HRMS (ESI) for C 48 H 53 N 10 O 5 [M + H] + , calcd: 849.4195, found: 849.4192.

[0200] Example 7: Preparation of Compound zlc - 5 - 15

[0201]

[0202] The synthesis method refers to Example 1.

[0203] 1 H NMR (500 MHz, DMSO-d 6 ) δ 11.09 (s, 1H), 8.29 (d, J = 2.3 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.60 (d, J = 8.9 Hz, 1H), 7.49 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.03 - 6.98 (m, 4H), 6.82 (s, 1H), 6.67 (d, J = 8.4 Hz, 1H), 6.46 (d, J = 8.9 Hz, 1H), 5.62 - 5.60 (m, 1H), 5.08 - 5.04 (m, 1H), 4.28 - 4.23 (m, lH), 4.14 (d, J = 5.2 Hz, 4H), 3.92 (s, 2H), 3.39 (s, 1H), 3.23 (s, 4H), 2.91 - 2.84 (m, 1H), 2.64 - 2.51 (m, 6H), 2.02 - 2.01 (m, 1H), 1.90 (d, J = 11.6 Hz, 2H), 1.77 - 1.75 (m, 2H), 1.35 - 1.22 (m, 3H), 1.17 - 1.05 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 172.87, 170.17, 167.51, 167.20, 159.26, 156.83, 154.94, 153.14, 150.06, 141.37, 133.86, 131.57, 128.05, 126.72, 126.27, 124.91, 119.17, 116.92, 115.45, 114.27, 104.56, 94.00, 55.06, 54.19, 52.91, 49.16, 48.74, 47.40, 43.49, 31.33, 31.01, 30.23, 22.23, 14.13. HRMS (ESI) for C 46 H 49 N 10 O 5 [M + H] + , calcd: 821.3882, found: 821.3884.

[0204] Example 8: Preparation of Compound zlc - 5 - 20

[0205]

[0206] The synthesis method refers to Example 1.

[0207] 1 1H NMR (500 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 8.29 (d, J = 1.8 Hz, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.6 Hz, 1H), 7.48 (d, J = 5.9 Hz, 1H), 7.28 - 7.25 (m, 2H), 7.19 - 7.15 (m, 3H), 7.01 - 6.98 (m, 5H), 6.86 (d, J = 8.6 Hz, 1H), 6.47 (d, J = 8.9 Hz, 1H), 5.59 (t, J = 5.7 Hz, 1H), 5.06 (dd, J = 12.8, 5.3 Hz, 1H), 4.29 - 4.23 (m, 1H), 4.15 (d, J = 5.7 Hz, 2H), 3.75 - 3.72 (m, 1H), 3.61 - 3.58 (m, 1H), 3.50 - 3.38 (m, 2H), 3.29 - 3.22 (m, 5H), 3.05 - 2.98 (m, lH), 2.91 - 2.85 (m, 1H), 2.69 - 2.61 (m, 4H), 2.61 - 2.52 (m, 2H), 2.31 - 2.27 (m, 1H), 2.02 - 2.00 (m, 1H), 1.91 - 1.89 (m, 3H), 1.77 (d, J = 10.5 Hz, 2H), 1.33 - 1.27 (m, 2H), 1.13 - 1.06 (m, 2H). 13 13C NMR (126 MHz, DMSO) δ 172.89, 170.22, 167.75, 167.29, 159.27, 156.85, 153.15, 151.81, 150.13, 141.38, 134.04, 131.56, 128.29, 126.72, 126.27, 124.96, 119.18, 115.80, 115.48, 115.25, 105.63, 94.01, 63.55, 52.92, 52.03, 51.35, 48.71, 47.64, 47.05, 43.50, 31.34, 31.03, 30.24, 28.89, 22.29. HRMS (ESI) for C 47 H 51 N 10 O 5 [M + H] + , calcd: 835.4038, found: 835.4036.

[0208] Example 9: Preparation of Compound zlc - 5 - 31

[0209]

[0210] The synthesis method refers to Example 1.

[0211] 1 H NMR (500 MHz, DMSO-d 6 ) δ 11.10 (s, 1H), 8.30 (d, J = 1.8 Hz, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.54 - 7.46 (m, 1H), 7.34 (s, 1H), 7.28 - 7.24 (m, 3H), 7.19 - 7.15 (m, 3H), 7.05 - 7.00 (m, 4H), 6.47 (d, J = 8.9 Hz, 1H), 5.60 - 5.59 (m, 1H), 5.07 (dd, J = 12.8, 5.3 Hz, 1H), 4.29 - 4.24 (m, 1H), 4.15 (d, J = 5.5 Hz, 2H), 4.08 (d, J = 12.6 Hz, 2H), 3.71 (s, 2H), 3.61 (s, 2H), 3.49 (s, 1H), 3.23 (s, 2H), 3.17 (s, 2H), 3.11 - 3.02 (m, 3H), 2.92 - 2.85 (m, 1H), 2.60 - 2.54 (m, 2H), 2.03 - 2.00 (m, 1H), 1.90 (d, J = 10.5 Hz, 2H), 1.78 - 1.73 (m, 4H), 1.67 - 1.60 (m, 2H), 1.33 - 1.27 (m, 2H), 1.13 - 1.06 (m, 2H). 13 C NMR (151 MHz, DMSO) δ 172.81, 172.34, 170.10, 167.60, 166.96, 159.24, 156.77, 154.82, 153.09, 149.92, 141.30, 134.05, 131.60, 128.70, 128.02, 126.70, 126.24, 125.03, 119.11, 117.64, 115.88, 107.85, 93.99, 52.92, 48.73, 48.41, 47.84, 46.72, 44.62, 43.48, 41.07, 36.80, 31.30, 30.98, 30.20, 27.42, 22.19, 14.09. HRMS (ESI) for C 49 H 53 N 10 O 6 [M + H] + , calcd: 877.4144, found: 877.4139.

[0212] Example 10: Preparation of Compound zlc-5-103

[0213]

[0214] The synthesis method refers to Example 1.

[0215] 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 8.30 - 8.28 (m, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.6 Hz, 1H), 7.47 (s, 1H), 7.36 (s, 1H), 7.28 - 7.25 (m, 3H), 7.18 - 7.15 (m, 3H), 6.97 (d, J = 8.6 Hz, 2H), 6.49 (d, J = 8.7 Hz, 2H), 6.46 (d, J = 9.0 Hz, 1H), 5.51 (t, J = 5.7 Hz, 1H), 5.07 (dd, J = 12.8, 5.4 Hz, 1H), 4.28 - 4.24 (m, 1H), 4.15 (d, J = 5.9 Hz, 2H), 3.98 (t, J = 7.1 Hz, 2H), 3.70 - 3.68 (m, 2H), 3.50 - 3.42 (m, 5H), 3.38 - 3.37 (m, 2H), 2.91 - 2.85 (m, 1H), 2.60 - 2.52 (m, 3H), 2.02 - 2.00 (m, 1H), 1.90 (d, J = 10.2 Hz, 2H), 1.76 (d, J = 10.6 Hz, 2H), 1.33 - 1.23 (m, 4H), 1.12 - 1.06 (m, 2H). 13 C NMR (151 MHz, DMSO) δ 173.28, 170.55, 168.03, 167.45, 159.72, 157.38, 155.70, 153.58, 151.37, 141.79, 134.31, 131.92, 128.50, 127.19, 127.15, 126.71, 125.37, 119.59, 118.93, 118.37, 112.19, 108.49, 94.46, 56.10, 54.60, 53.37, 49.25, 49.10, 47.17, 43.94, 31.78, 31.45, 30.67, 22.65. HRMS (ESI) for C 46 H 49 N 10 O 5 [M + H] + , calcd: 821.3882, found: 821.3884.

[0216] Example 11: Preparation of Compound zlc-5-104

[0217]

[0218] The synthesis method refers to Example 1.

[0219] 1 H NMR(600MHz, DMSO-d 6 )δ11.07(s, 1H), 8.29(d, J = 2.3Hz, 1H), 7.63(d, J = 8.5Hz, 1H), 7.60(d, J = 8.3Hz, 1H), 7.48(s, 1H), 7.28 - 7.25(m, 3H), 7.21 - 7.15(m, 4H), 6.97(d, J = 8.5Hz, 2H), 6.67(d, J = 8.4Hz, 2H), 6.47(d, J = 8.9Hz, 1H), 5.53(s, 1H), 5.05(dd, J = 12.8, 5.5Hz, 1H), 4.29 - 4.25(m, 1H), 4.15(d, J = 5.8Hz, 2H), 4.02(d, J = 12.3Hz, 2H), 3.41(m, 3H), 3.10(m, 2H), 2.96 - 2.84(m, 5H), 2.59 - 2.53(m, 4H), 2.43(s, 1H), 2.28 - 2.23(m, 2H), 2.01 - 1.97(m, 2H), 1.91 - 1.90(m, 2H), 1.79 - 1.75(m, 5H), 1.31 - 1.27(m, 2H), 1.24 - 1.23(m, 2H), 1.13 - 1.09(m, 2H).HRMS(ESI)for C 51 H 57 N 10 O 5 [M + H] + , calcd: 889.4508, found: 889.4515.

[0220] Example 12: Preparation of Compound zlc-5-105

[0221]

[0222] The synthesis method refers to Example 1.

[0223] 1 H NMR(600MHz, DMSO-d 6)δ 11.08 (s, 1H), 8.29 (d, J = 2.3 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.48 (s, 1H), 7.29 - 7.25 (m, 3H), 7.21 (dd, J = 8.7, 2.1 Hz, 1H), 7.18 - 7.14 (m, 3H), 6.95 (d, J = 8.6 Hz, 2H), 6.47 - 6.44 (m, 3H), 5.47 (t, J = 5.5 Hz, 1H), 5.06 (dd, J = 12.8, 5.5 Hz, 1H), 4.27 - 4.23 (m, 1H), 4.14 (d, J = 5.9 Hz, 2H), 4.01 (d, J = 12.9 Hz, 2H), 3.90 (s, 4H), 3.30 (s, 4H), 2.95 - 2.91 (m, 2H), 2.91 - 2.85 (m, 1H), 2.60 - 2.52 (m, 2H), 2.27 (s, 1H), 2.02 - 2.00 (m, 1H), 1.90 (d, J = 12.1 Hz, 2H), 1.75 (d, J = 11.8 Hz, 4H), 1.30 - 1.24 (m, 4H), 1.17 - 1.11 (m, 2H), 1.08 - 1.06 (m, 2H). 13 C NMR (151 MHz, DMSO) δ 173.29, 170.59, 168.11, 167.43, 159.71, 157.35, 155.40, 153.57, 151.12, 141.74, 134.52, 131.89, 128.51, 125.49, 119.59, 118.02, 117.81, 112.31, 108.16, 94.46, 70.25, 64.88, 62.14, 53.35, 49.20, 47.71, 43.94, 34.71, 31.77, 31.45, 30.65, 29.90, 22.66. HRMS (ESI) for C 50 H 55 N 10 O 5 [M + H] + , calcd: 875.4351, found: 875.4354.

[0224] Example 13: Preparation of Compound zlc - 5 - 106

[0225]

[0226] The synthesis method refers to Example 1.

[0227] 11H NMR (600 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.64 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.7 Hz, 1H), 7.48 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.02 - 6.97 (m, 4H), 6.90 (d, J = 1.8 Hz, 1H), 6.81 (dd, J = 8.6, 2.1 Hz, 1H), 6.47 (d, J = 8.9 Hz, 1H), 5.57 (t, J = 5.6 Hz, 1H), 5.05 (dd, J = 12.7, 5.5 Hz, 1H), 4.28 - 4.23 (m, 1H), 4.15 (d, J = 5.9 Hz, 2H), 3.59 - 3.57 (m, 1H), 3.52 - 3.49 (m, 2H), 3.43 - 3.37 (m, 2H), 3.21 (s, 4H), 3.17 - 3.14 (m, 1H), 2.91 - 2.85 (m, 1H), 2.68 - 2.63 (m, 1H), 2.59 - 2.56 (m, 5H), 2.41 (d, J = 6.9 Hz, 2H), 2.18 - 2.14 (m, 1H), 2.02 - 1.99 (m, 1H), 1.90 (d, J = 10.4 Hz, 2H), 1.81 - 1.76 (m, 3H), 1.31 - 1.27 (m, 2H), 1.13 - 1.07 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 173.29, 170.63, 168.20, 167.71, 159.72, 157.29, 153.57, 152.35, 150.61, 141.80, 134.49, 131.98, 128.50, 127.16, 126.71, 125.44, 119.59, 115.96, 115.84, 115.73, 105.93, 94.46, 61.37, 53.56, 53.39, 52.68, 49.15, 48.14, 47.69, 43.94, 35.99, 31.78, 31.47, 30.67, 29.74, 22.73. HRMS (ESI) for C 48 H 53 N 10 O 5 [M + H] + , calcd: 849.4195, found: 849.4200.

[0228] Example 14: Preparation of Compound zlc - 6 - 1

[0229]

[0230] The synthesis method refers to Example 1.

[0231] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.64 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.48 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.02 - 6.97 (m, 4H), 6.91 (d, J = 1.7 Hz, 1H), 6.82 (dd, J = 8.6, 2.0 Hz, 1H), 6.47 (d, J = 8.9 Hz, 1H), 5.58 - 5.56 (m, 1H), 5.05 (dd, J = 12.7, 5.5 Hz, 1H), 4.28 - 4.24 (m, 1H), 4.15 (d, J = 5.8 Hz, 2H), 3.60 - 3.57 (m, 1H), 3.53 - 3.49 (m, 2H), 3.43 - 3.39 (m, 2H), 3.21 (s, 4H), 3.17 - 3.15 (m, 1H), 2.91 - 2.85 (m, 1H), 2.67 - 2.64 (m, 1H), 2.59 - 2.52 (m, 5H), 2.42 (d, J = 7.0 Hz, 2H), 2.18 - 2.15 (m, 1H), 2.02 - 1.96 (m, 1H), 1.90 (d, J = 10.3 Hz, 2H), 1.81 - 1.76 (m, 3H), 1.31 - 1.29 (m, 2H), 1.13 - 1.07 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 173.29, 170.63, 168.20, 167.71, 159.72, 157.29, 153.58, 152.35, 150.61, 141.80, 134.49, 131.98, 128.50, 127.16, 126.71, 125.45, 119.59, 115.96, 115.84, 115.73, 105.93, 94.46, 61.37, 53.57, 53.39, 52.68, 49.15, 48.14, 47.69, 43.94, 35.99, 31.78, 31.46, 30.67, 29.74, 22.56. HRMS (ESI) for C 48 H 53 N 10 O 5 [M + H] +, calcd: 849.4195, found: 849.4191.

[0232] Example 15: Preparation of Compound zlc-6-35

[0233]

[0234] The synthesis method refers to Example 1.

[0235] 1 H NMR(600MHz, DMSO-d 6 ) δ11.08(s, 1H), 8.29(d, J = 2.2Hz, 1H), 7.68(d, J = 8.5Hz, 1H), 7.60(d, J = 8.9Hz, 1H), 7.47(s, 1H), 7.36 - 7.34(m, 1H), 7.28 - 7.25(m, 3H), 7.18 - 7.14(m, 3H), 6.95(d, J = 8.5Hz, 2H), 6.47 - 6.45(m, 3H), 5.48 - 5.47(m, 1H), 5.07(dd, J = 12.8, 5.4Hz, 1H), 4.27 - 4.23(m, 1H), 4.14(d, J = 5.9Hz, 2H), 3.97(m, 2H), 3.54 - 3.50(m, 2H), 3.45(s, 4H), 3.00 - 2.95(m, 1H), 2.64(d, J = 7.3Hz, 2H), 2.61 - 2.57(m, 2H), 2.54 - 2.52(m, 5H), 2.03 - 1.99(m, 1H), 1.90(d, J = 10.4Hz, 2H), 1.75(d, J = 11.6Hz, 2H), 1.31 - 1.27(m, 2H), 1.11 - 1.05(m, 2H). 13 C NMR(151MHz, DMSO) δ172.82, 170.09, 167.57, 166.99, 159.25, 156.92, 155.24, 153.11, 150.90, 141.29, 133.87, 131.44, 128.05, 126.70, 126.49, 124.91, 119.13, 118.35, 117.85, 111.48, 107.93, 94.00, 56.11, 52.87, 52.34, 48.78, 46.88, 43.48, 31.30, 30.98, 30.20, 29.03, 27.22. HRMS(ESI) for C 47 H 51 N 10 O 5 [M + H] +, calcd: 835.4038, found: 835.4045.

[0236] Example 16: Preparation of Compound zlc-6-37

[0237]

[0238] The synthesis method refers to Example 1.

[0239] 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.47 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.14 (m, 3H), 6.99 - 6.94 (m, 4H), 6.79 (s, 1H), 6.65 - 6.63 (dd, J = 8.4, 1.9 Hz, 1H), 6.47 - 6.46 (m, 1H), 5.54 (t, J = 5.7 Hz, 1H), 5.05 (dd, J = 12.8, 5.4 Hz, 1H), 4.28 - 4.22 (m, 1H), 4.14 (d, J = 5.9 Hz, 2H), 4.10 (s, 4H), 3.71 (d, J = 12.1 Hz, 2H), 3.36 (m, 2H), 2.90 - 2.84 (m, 1H), 2.67 - 2.63 (m, 2H), 2.59 - 2.56 (m, 1H), 2.56 - 2.52 (m, 2H), 2.48 - 2.46 (m, 2H), 2.02 - 1.98 (m, 1H), 1.90 (d, J = 10.3 Hz, 2H), 1.76 (d, J = 11.9 Hz, 4H), 1.45 - 1.38 (m, 1H), 1.33 - 1.26 (m, 2H), 1.24 - 1.17 (m, 4H), 1.13 - 1.06 (m, 2H). 13¹³C NMR (151 MHz, DMSO) δ 173.28, 170.57, 167.96, 167.64, 159.72, 157.30, 155.34, 153.57, 151.04, 141.79, 134.26, 131.92, 128.50, 128.23, 127.15, 126.71, 125.28, 119.59, 117.46, 116.27, 114.92, 105.17, 94.46, 64.77, 61.69, 53.38, 49.19, 48.61, 43.94, 34.69, 31.77, 31.45, 30.67, 22.67. HRMS (ESI) for C 50 H 55 N 10 O 5 [M+H] + , calcd: 875.4351, found: 875.4349.

[0240] Example 17: Preparation of Compound zlc-6-38

[0241]

[0242] The synthesis method refers to Example 1.

[0243] 1 ¹H NMR (600 MHz, DMSO-d 6 ) δ 11.14 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.92 (d, J = 2.9 Hz, 3H), 7.60 (d, J = 8.9 Hz, 1H), 7.47 (s, 1H), 7.28 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.02 - 6.99 (s, 4H), 6.46 (d, J = 8.9 Hz, lH), 5.57 (t, J = 5.8 Hz, 1H), 5.16 (dd, J = 12.9, 5.4 Hz, 1H), 4.28 - 4.23 (m, 1H), 4.14 (d, J = 6.0 Hz, 2H), 3.68 (s, 2H), 3.39 - 3.36 (m, 1H), 3.27 - 3.22 (m, 4H), 2.92 - 2.86 (m, 1H), 2.72 - 2.69 (m, 4H), 2.62 - 2.59 (m, 1H), 2.56 - 2.53 (m, 1H), 2.08 - 2.04 (m, 1H), 1.93 - 1.87 (m, 2H), 1.76 (d, J = 11.4 Hz, 2H), 1.32 - 1.26 (m, 2H), 1.12 - 1.06 (m, 2H). 1313C NMR (151 MHz, DMSO) δ 172.75, 169.76, 166.50, 166.42, 156.80, 153.11, 150.14, 141.33, 137.58, 131.83, 131.52, 130.28, 128.77, 128.32, 128.03, 126.70, 126.24, 125.74, 123.83, 119.13, 115.55, 93.99, 90.49, 83.72, 52.91, 51.46, 49.12, 47.62, 46.75, 43.47, 31.31, 30.92, 30.20, 21.93. HRMS (ESI) for C 46 H 46 N 9 O 5 [M + H] + , calcd: 804.3616, found: 804.3623.

[0244] Example 18: Preparation of Compound zlc-6-42

[0245]

[0246] The synthesis method refers to Example 1.

[0247] 1 1H NMR (600 MHz, DMSO-d 6)δ11.08(s, 1H), 8.29(d, J = 2.4, 1H), 7.68(d, J = 8.5Hz, 1H), 7.60(d, J = 9.0Hz, 1H), 7.47(s, lH), 7.34(d, J = 2.2Hz, 1H), 7.28 - 7.25(m, 3H), 7.18 - 7.15(m, 3H), 6.99 - 6.96(m, 4H), 6.47(d, J = 8.9Hz, 1H), 5.55(t, J = 6.0Hz, 1H), 5.07(dd, J = 12.8, 5.4Hz, 1H), 4.28 - 4.23(m, 1H), 4.15(d, J = 6.1Hz, 2H), 3.74(d, J = 11.8Hz, 2H), 3.44(s, 4H), 3.42 - 3.36(m, 4H), 2.90 - 2.85(m, 1H), 2.72 - 2.68(m, 2H), 2.61 - 2.54(m, 2H), 2.23(d, J = 7.2Hz, 2H), 2.03 - 1.99(m, 1H), 1.90(d, J = 11.6Hz, 2H), 1.84(d, J = 12.1Hz, 2H), 1.77 - 1.72(m, 3H), 1.31 - 1.28(m, 2H), 1.25 - 1.20(m, 4H), 1.13 - 1.07(m, 2H). 13 C NMR(151MHz, DMSO)δ172.83, 170.10, 167.58, 167.00, 159.26, 156.85, 155.27, 153.11, 150.62, 141.33, 133.87, 131.46, 128.04, 127.74, 126.70, 126.25, 124.92, 119.14, 118.32, 117.78, 115.80, 107.89, 94.00, 63.74, 52.91, 52.76, 48.78, 48.12, 46.96, 43.48, 32.44, 31.32, 30.99, 30.32, 30.21, 22.19.HRMS(ESI)for C 49 H 55 N 10 O 5 [M + H] + , calcd: 863.4351, found: 863.4357.

[0248] Example 19: Preparation of Compound zlc - 6 - 101

[0249]

[0250] The synthesis method refers to Example 1.

[0251] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 8.29 (d, J = 2.1 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.7 Hz, 1H), 7.48 (s, 1H), 7.32 (s, 1H), 7.28 - 7.23 (m, 3H), 7.18 - 7.15 (m, 3H), 6.99 (s, 4H), 6.50 - 6.43 (m, 1H), 5.54 - 5.52 (m, 1H), 5.06 (dd, J = 12.8, 5.5 Hz, 1H), 4.28 - 4.24 (m, 1H), 4.15 (d, J = 5.8 Hz, 2H), 3.51 (s, 6H), 3.22 (s, 4H), 2.91 - 2.85 (m, 1H), 2.60 - 2.57 (m, 2H), 2.03 - 1.99 (m, 1H), 1.91 (d, J = 10.6 Hz, 2H), 1.77 (d, J = 10.4 Hz, 2H), 1.63 - 1.59 (m, 8H), 1.33 - 1.28 (m, 2H), 1.14 - 1.07 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 172.83, 170.14, 167.67, 167.00, 159.25, 156.84, 154.97, 153.11, 141.31, 134.04, 131.46, 128.05, 127.66, 126.70, 126.26, 125.00, 119.13, 117.35, 115.47, 107.50, 94.00, 69.79, 52.90, 48.74, 43.48, 42.89, 34.82, 34.12, 31.31, 30.99, 30.21, 29.14, 22.21. HRMS (ESI) for C 48 H 52 N 9 O 5 [M + H] + , calcd: 834.4086, found: 834.4073.

[0252] Example 20: Preparation of Compound zlc - 6 - 102

[0253]

[0254] The synthesis method refers to Example 1.

[0255] 1 1H NMR (600 MHz, DMSO-d6 ) δ 11.06 (s, 1H), 8.30 (d, J = 2.2 Hz, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.48 - 7.45 (m, 1H), 7.28 - 7.26 (m, 2H), 7.19 - 7.15 (m, 3H), 7.02 - 6.99 (m, 4H), 6.96 (s, 1H), 6.83 (dd, J = 8.6, 1.6 Hz, 1H), 6.47 (d, J = 8.9 Hz, 1H), 5.52 (t, J = 5.6 Hz, 1H), 5.07 - 5.04 (m, 1H), 4.29 - 4.24 (m, 1H), 4.15 (d, J = 5.9 Hz, 2H), 3.53 - 3.50 (m, 2H), 3.37 (m, 4H), 3.24 - 3.21 (m, 1H), 2.91 - 2.85 (m, 1H), 2.61 - 2.52 (m, 4H), 2.02 - 1.98 (m, 1H), 1.97 - 1.95 (m, 2H), 1.91 (d, J = 10.7 Hz, 2H), 1.77 (d, J = 10.3 Hz, 2H), 1.73 - 1.66 (m, 4H), 1.33 - 1.27 (m, 2H), 1.13 - 1.07 (m, 2H). 13 C NMR (151 MHz, DMSO) δ 172.83, 170.16, 167.74, 167.26, 159.25, 156.82, 153.12, 152.06, 150.42, 141.28, 134.02, 131.48, 128.05, 127.83, 126.70, 126.27, 124.94, 119.13, 115.80, 115.54, 115.22, 105.58, 94.00, 57.28, 52.90, 48.69, 46.32, 45.44, 43.49, 35.02, 34.06, 31.31, 31.00, 30.22, 22.26. HRMS (ESI) for C 47 H 50 N 9 O 5 [M + H] + , calcd: 820.3929, found: 820.3933.

[0256] Example 21: Preparation of Compound zlc - 6 - 103

[0257]

[0258] The synthesis method refers to Example 1.

[0259] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 8.30 (d, J = 2.2 Hz, 1H), 7.65 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 8.1 Hz, 1H), 7.48 (s, 1H), 7.28 - 7.26 (m, 2H), 7.19 - 7.15 (m, 3H), 7.06 - 7.03 - 6.99 (m, 4H), 6.80 (d, J = 1.8 Hz, 1H), 6.67 (dd, J = 8.4, 2.0 Hz, 1H), 6.47 (d, J = 8.8 Hz, 1H), 5.56 (t, J = 5.6 Hz, 1H), 5.05 (dd, J = 12.8, 5.5 Hz, 1H), 4.29 - 4.23 (m, 1H), 4.15 (d, J = 5.8 Hz, 1H), 3.83 (m, 4H), 3.43 - 3.41 (m, 2H), 3.22 (m, 3H), 2.91 - 2.85 (m, 1H), 2.59 - 2.53 (m, 2H), 2.02 - 1.89 (m, 1H), 1.91 - 1.89 (m, 6H), 1.77 (d, J = 10.1 Hz, 2H), 1.33 - 1.27 (m, 2H), 1.13 - 1.07 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 172.83, 170.12, 167.53, 167.21, 159.26, 156.83, 155.20, 153.12, 150.21, 141.33, 133.86, 131.53, 128.04, 126.71, 126.26, 124.87, 122.51, 119.14, 116.69, 115.95, 114.18, 104.42, 94.00, 60.78, 52.92, 48.73, 45.32, 43.49, 34.65, 34.21, 31.32, 30.99, 30.21, 22.23, 20.45. HRMS (ESI) for C 46 H 48 N 9 O 5 [M + H] + , calcd: 806.3773, found: 806.3767.

[0260] Example 22: Preparation of Compound zlc - 7 - 36

[0261]

[0262] The synthesis method refers to Example 1.

[0263] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.60 - 7.58 (m, 2H), 7.51 - 7.44 (m, 1H), 7.29 - 7.25 (m, 2H), 7.18 - 7.15 (m, 3H), 7.01 - 6.97 (m, 4H), 6.91 (d, J = 7.6 Hz, 1H), 6.46 (d, J = 8.9 Hz, 1H), 5.57 (t, J = 5.6 Hz, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.30 - 4.24 (m, 3H), 4.15 (d, J = 5.8 Hz, 2H), 3.87 - 3.85 (m, 2H), 3.53 - 3.44 (m, 1H), 3.19 (s, 4H), 3.04 - 3000 (m, 1H), 2.90 - 2.84 (m, 1H), 2.67 (d, J = 6.8 Hz, 2H), 2.61 - 2.51 (m, 6H), 2.03 - 1.99 (m, 1H), 1.90 (d, J = 10.5 Hz, 2H), 1.76 (d, J = 10.6 Hz, 2H), 1.31 - 1.27 (m, 2H), 1.12 - 1.07 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 172.80, 170.02, 166.84, 166.43, 159.26, 156.83, 154.43, 153.11, 152.79, 144.21, 144.13, 141.34, 131.52, 129.38, 128.23, 128.04, 126.70, 119.14, 118.30, 115.43, 111.33, 111.19, 107.95, 94.00, 61.57, 57.75, 52.93, 52.83, 48.94, 47.64, 43.48, 31.32, 30.97, 30.21, 27.85, 22.17, 22.08, 13.99. HRMS (ESI) for C 47 H 50 FN 10 O 5 [M + H] + , calcd: 853.3944, found: 853.3956.

[0264] Example 23: Test of the CDK12 / 13 Degradation Activity of Representative Compounds in MDA-MB-231 Cells

[0265] Experimental method: The cell line MDA-MB-231 was purchased from the American Type Culture Collection (ATCC). Detection was performed using conventional Western Blot as follows: MDA-MB-231 cells were seeded in a 12-well plate at a density of 4×10 5 / mL, 1 mL per well, and cultured overnight in an incubator for cell attachment. Subsequently, a certain concentration of the compound was added and the cells were treated for 15 h. The cells were lysed with lysis buffer and protein samples were collected. An appropriate amount of the sample was subjected to SDS-PAGE electrophoresis. After electrophoresis, the proteins were transferred to a polyvinylidene difluoride (PVDF) membrane using a wet electrotransfer system. The electrotransferred PVDF membrane was cut into several required strips, and then placed in a blocking solution (5% skim milk diluted in TBS containing 0.1% Tween 20) and blocked at room temperature for 1.5 h. The primary antibodies against the corresponding target proteins were diluted in TBS containing 0.1% Tween 20 according to the dilution requirements of different antibodies, such as 1:1000 (CST), 1:3000 (Proteintech, Bethyl), 1:200 (Santacruz), to prepare the primary antibody solutions. The blocked membranes were placed in the corresponding primary antibody solutions and incubated at 4 °C for 12 - 14 h. Subsequently, they were washed three times with TBS containing 0.1% Tween 20, 5 min each time. The washed membranes were placed in a secondary antibody solution (horseradish peroxidase-labeled goat anti-rabbit IgG, diluted 1:2000 in TBS containing 0.1% Tween 20) and reacted at room temperature for 2 h. After washing the membranes three times as above, they were developed with StarSignal plus reagent and Omni-ECL reagent and photographed with an Amersham Imager 800 system.

[0266] Figure 1 The results showed that some compounds could effectively degrade the protein levels of CDK12 and CDK13 in cells, such as compounds ZLC4-91, ZLC4-93, ZLC5-6, ZLC5-11, ZLC5-31, ZLC5-103, ZLC5-104, ZLC5-106, ZLC6-1, ZLC6-37, and ZLC6-42, etc. Compound ZLC5-11 could degrade the CDK12 and CDK13 proteins in MDA-MB-231 cells in a time-dependent manner. After treating MDA-MB-231 cells at a concentration of 1 μM for 15 h, the CDK12 and CDK13 proteins in the cells were almost completely degraded. In addition, ZLC5-11 was found to have a time-dependent degradation effect on the CDK12 and CDK13 chaperone protein Cyclin K. After 15 h, the Cyclin K protein in the cells had been almost completely degraded.

[0267] We further determined the degradation of CDK12 and CDK13 proteins in MDA-MB-231 cells by representative compounds ZLC5-6, ZLC5-31, ZLC5-11, ZLC6-37, ZLC6-42, and ZLC5-15 at different concentrations ( Figure 2 ). The experimental results showed that these representative compounds could degrade CDK12 and CDK13 proteins in MDA-MB-231 cells in a dose-dependent manner, and the target proteins could be effectively degraded at a concentration of 1 μM ( Figure 2 ).

[0268] Figure 1 , (A) WB results of CDK12 protein degradation after treating MDA-MB-231 cells with some compounds at a concentration of 0.1 μM for 15 hours. (B) WB results of CDK13 protein degradation after treating MDA-MB-231 cells with some compounds at a concentration of 0.1 μM for 15 hours. (C) Using DMSO in the control group as a reference standard, the protein levels were quantified by the gray values of the bands in the WB result images (Figure A and Figure B). (D) Compound ZLC5-11 degraded CDK12 and CDK13 proteins in MDA-MB-231 cells in a time-dependent manner, and the target proteins could be significantly degraded at 15 h.

[0269] Figure 2 , WB measurement results of intracellular CDK12 and CDK13 protein levels after treating MDA-MB-231 cells with representative compounds ZLC5-6, ZLC5-31, ZLC5-11, ZLC6-37, ZLC6-42, and ZLC5-15 at different concentrations for 15 hours.

[0270] Example 24: Study on the proliferation inhibitory activity of compounds against triple-negative breast cancer cell MDA-MB-231

[0271] The cell proliferation inhibitory activity of the compounds was determined using the Cell Counting Kit-8 Cell Viability Assay (Selleck.cn). The cells were seeded in a 384-well plate containing its corresponding medium ( PS 3701), and incubated in an incubator at 37 °C with 5% CO 2 . After incubating overnight, the high-concentration stock solutions of each compound to be tested were prepared; using 650 Liquid Handler the compounds were added to the 384-well plate at a certain concentration gradient. After drug administration, the cell plate was placed in an incubator at 37 °C with 5% CO 2 and incubated for 5 days. Subsequently, using (Thermo ScientificTM ) Add 5 μL of CCK-8 reagent to each well of a 384-well plate, and then put the plate back into the incubator for incubation for 1.5 - 2 h. After incubation, take out the plate, centrifuge to remove air bubbles, and then use Measure the absorbance values of each well at 450 nm and 650 nm, and use GraphPad Prism software (GraphPad Software Inc) to process and analyze the data.

[0272] Figure 3 The results showed that compounds capable of effectively degrading the protein levels of CDK12 and CDK13, such as ZLC4-77, ZLC4-91, ZLC5-6, ZLC5-31, and ZLC6-1, could effectively inhibit the growth of triple-negative breast cancer cells MDA-MB-231, and the IC 50 values for the MDA-MB-231 cell line were 610.4 nM, 622.5 nM, 497.9 nM, 389.0 nM, and 288.6 nM, respectively.

[0273] Example 25: In vivo pharmacokinetic experiments of representative compounds zlc-4-91 and zlc-4-93 of cell cycle-dependent protein kinase 12 / 13 (CDK12 / 13) degrading agents

[0274] Pharmacokinetic and bioavailability tests in rats. SD rats were administered by single oral (10 mg / kg), intravenous (2.5 mg / kg), and intraperitoneal (10 mg / kg) injection. After administration, animal blood samples were collected at appropriate time points, anticoagulated with heparin, centrifuged at 8000 rpm for 6 minutes, and the supernatant was taken and stored at -20 °C for HPLC-MS analysis. The blood samples were precipitated with acetonitrile for protein at 12000 rpm * 10 min, and the supernatant was used for HPCL-MS analysis. The data were fitted with parameters using DAS2.0 to obtain compartment model and non-compartment model parameters respectively. The oral bioavailability of the compound was calculated according to the area under the plasma concentration-time curve (AUC) data. The results are shown in Table 1. The experimental results showed (Table 1) that the representative compounds zlc-4-91 and zlc-4-93 had excellent oral pharmacokinetic properties.

[0275] Table 1. Pharmacokinetic experimental results of compound zlc-4-93

[0276]

[0277] C max refers to the maximum plasma concentration, T 1 / 2 is the half-life, CL refers to the clearance rate, and F refers to the bioavailability.

[0278] All documents mentioned in this invention are cited in this application as references, just as if each document was cited separately as a reference. The above-described embodiments only express several implementation manners of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope recorded in this specification.

Claims

1. A compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a prodrug molecule thereof: Wherein, Z is: CH 2 or CO; V is selected from the following group: X and Y are each independently selected from the group consisting of: N, CH or CR 3 ; Among them, R 3 is selected from the following group: halogen, cyano, hydroxyl, amino, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 3 alkoxy, halo-C 1 -C 3 alkoxy, C 3 -C 8 cycloalkyl, or 3-8 membered heterocyclic group; B is selected from the following group: NH, O, CO, CH 2 ; wherein each U and W is independently selected from the group: N or CH; each m, p, m', p' is independently 0, 1, 2 or 3; Ring A is selected from the following group: Wherein, Q and W are each independently selected from: CH, N; R′ is independently selected from: hydrogen, halogen, cyano, hydroxy, substituted hydroxy, amino, substituted amino, C 1 -C 5 -alkyl, halo-C 1 -C 5 -alkyl, C 1 -C 5 -alkoxy, halo-C 1 -C 5 -alkoxy, C 3 -C 8 -cycloalkyl, and when Q and W are independently selected from CH, R′ may be a substituent on Q and W; D, E, F, and G are each independently selected from the group consisting of: CH, N, CR 6 ; wherein, R 6 is selected from the group consisting of: halogen, trifluoromethyl, hydroxyl, cyano, amino, methyl, methoxy, trifluoromethoxy; and when any one of D, E, F, and G is CH, this atom can serve as a connection site with the linker, and in this case, the said D, E, F, or G is C; R 1 selected from the group consisting of: H, -NHR 7 , -OR 7 , -(C(R 9 )R 8 )R 7 ; Among them, R 7 is -R 10 , -CH 2 R 10 or -(CH 2 ) 2 R 10 ; R 8 、R 9 are each independently selected from the group consisting of: hydrogen, halogen, cyano, methyl, halo-methyl, methoxy, halo-methoxy, ethyl, halo-ethyl, ethoxy, halo-ethoxy, hydroxy, amino, a 3- to 8-membered heterocyclic ring containing 1, 2 or 3 heteroatoms, and the heteroatoms are selected from O, S or N; or R 8 , R 9 and the C atom connected thereto together form a 3- to 7-membered heterocycle; R 10 Selected from the group consisting of: 1) Cyano, C 1 -C 5 alkyl, halo C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 3 ~C 10 cycloalkyl, substituted or unsubstituted 5- to 12-membered aromatic ring, substituted or unsubstituted 3- to 12-membered heterocyclic ring; 2) where Q 1 , Q 2 , Q 3 , Q 4 , Q 5 are each independently selected from: CH, N or CR 11 ; Each R 11 is independently selected from the group consisting of: halogen, cyano, hydroxy, amino, nitro, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 4 alkoxy, halo-C 1 -C 4 alkoxy, C 3 -C 8 cycloalkyl; R 2 Selected from the group consisting of: H, C 1 -C 3 alkyl, When B is selected from the following group: NH, O, CO, CH 2 ; wherein each U and W is independently selected from the group: N or CH; each of m, p, m', p' is independently 0, 1, 2, or 3; when The Linker is: Wherein, R L1 、R L2 、R L3 、R L4 and R L5 are the same or different and each independently selected from the group consisting of substituted or unsubstituted groups: chemical bond, CH 2 、CHD、CD 2 、C=O、O、NH、SO、SO 2 、P=O、NHCO、NHSO 2 、OCH 2 、OCH 2 CH 2 、CH 2 OCH 2 、NHCH 2 、NMeCH 2 、NHCH 2 CH 2 、NMeCH 2 CH 2 、CH 2 NHCO、NHCOCH 2 、 R L6 is a ring and is arbitrarily selected from the following structures: Wherein each n is independently 0, 1, 2, 3, 4, 5 or 6; each r and m are independently 0, 1 or 2; each U and W are independently selected from the group consisting of: N or CH; p L1 、p L2 、p L3 、p L4 、p L5 and p L6 are each independently selected from 0, 1, 2, 3, 4, 5, or 6; or When B is where each of m, p, m', and p' is independently 0, 1, 2, or 3; when The Linker is: Wherein, R L1 、R L2 、R L3 、R L4 、R L5 and R L6 are the same or different and each independently selected from the group consisting of substituted or unsubstituted groups: chemical bond, CH 2 、CHD、CD 2 、C=O、O、NH、SO、SO 2 、P=O、NHCO、NHSO 2 、OCH 2 、OCH 2 CH 2 、CH 2 OCH 2 、NHCH 2 、NMeCH 2 、NHCH 2 CH 2 、NMeCH 2 CH 2 、CH 2 NHCO、NHCOCH 2 、 Wherein each n is independently 0, 1, 2, 3, 4, 5 or 6; each r and m are independently 0, 1 or 2; each U and W are independently selected from the group consisting of: N or CH; p L1 、p L2 、p L3 、p L4 、p L5 and p L6 are each independently selected from 0, 1, 2, 3, 4, 5, or 6.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, V is selected from the following group:

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, X and Y are each independently selected from the group consisting of: N, CH or CR 3 ; wherein, R 3 is selected from the group consisting of: halogen, cyano, hydroxy, amino, C 1 -C 3 alkyl, halo-C 1 -C 3 alkyl, C 1 -C 3 alkoxy, halo-C 1 -C 3 alkoxy.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, B is selected from the following group: NH, O, CO; Wherein each U and W are independently selected from the group consisting of: N or CH; each m, p, m', p' are independently 0, 1, 2 or 3.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, Ring A is selected from the following group: Wherein, W is selected from the group consisting of: CH, N; R′ is selected from: hydrogen, halogen, cyano, hydroxy, amino, C 1 -C 5 -alkyl, halo-C 1 -C 5 -alkyl, C 1 -C 5 -alkoxy, halo-C 1 -C 5 -alkoxy, C 3 -C 8 -cycloalkyl.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, D, E, F, and G are CH and CR 6 ; wherein, R 6 is selected from the group consisting of: halogen 7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, The described R 1 is -NHR 7 ; Among them, R 7 is -CH 2 R 10 ; The definition of R 10 is as described in claim 1. In another preferred example, the R 10 is selected from the group consisting of: cyano, C 1 -C 5 alkyl, halo-C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 3 -C 10 cycloalkyl, 4-7 membered heterocyclic group, C 6 -C 10 aryl; wherein the aryl is substituted by one or more CR 11 ; and the definition of R 11 is as described in claim 1.

8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, When B is selected from the following group: NH, O, CO: the Linker is selected from the following group: and when B is the Linker is selected from the following group: chemical bond, 9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, Having the structure shown in formula (II): Among them, B is selected from: Wherein each U and W are independently selected from the group consisting of: N or CH; each m, p, m', p' are independently 0, 1, 2 or 3; R' is optionally selected from: hydrogen, halogen, cyano, hydroxyl, amino; R 11 、R 6 Optionally selected from: hydrogen, halogen; X and Y are independently selected from: CH, N or CR 3 ; wherein, R 3 is optionally selected from: halogen, cyano, hydroxyl, amino; Z is selected from: CH 2 or CO.

10. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, Characterized in that, The compound is selected from the group consisting of:

11. A pharmaceutical composition, Characterized in that, Comprising (1) The compound according to claim 1 as an active ingredient, or a pharmaceutically acceptable salt, a stereoisomer thereof or a prodrug molecule thereof; and optionally (2) a pharmaceutically acceptable carrier.

12. The use of the compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, or the pharmaceutical composition according to claim 11, Characterized in that, For the preparation of a CDK12 / 13 protein kinase degrader.

13. Use of the compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, or the pharmaceutical composition according to claim 11, characterized in that, it is used for preparing a drug for preventing and / or treating a disease mediated by CDK12 / 13 serine / threonine protein kinase.

14. The use according to claim 13, characterized in that, the diseases mediated by CDK12 / 13 serine / threonine protein kinase are selected from the following group: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing's sarcoma, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, leukemia, histiocytic lymphoma, nasopharyngeal carcinoma, head and neck tumor, colon cancer, rectal cancer, glioma.

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