Novel CDK12 / 13 covalent inhibitor having a fused ring structure substituent or pharmaceutical composition thereof and use thereof

Abstract

The present invention relates to a preparation method for a novel CDK12 / 13 covalent inhibitor having a fused ring structure substituent or a pharmaceutical composition thereof and the use thereof. The novel CDK12 / 13 covalent inhibitor having a fused ring structure substituent has a structure shown as formula (I), and the compound can be used as a protein kinase inhibitor, can effectively and highly selectively inhibit the activity of CDK12 / 13 protein kinase, and can inhibit the proliferation, migration and invasion of a plurality of tumor cells.

Description

A novel CDK12 / 13 covalent inhibitor with cyclic structure substitution or its pharmaceutical composition and application Technical Field

[0001] The present invention belongs to the field of chemical medicine, and in particular relates to a novel CDK12 / 13 covalent inhibitor with a cyclic structure substitution or a pharmaceutical composition and application thereof. Background Art

[0002] Cyclin-dependent kinases (CDKs) are a class of serine / threonine kinases that play a crucial role in the control of cell division and regulate transcription in response to a variety of intracellular and extracellular signals. Twenty CDKs and 29 cyclins have been identified and reported in mammals. CDKs require binding to one or more of their corresponding cyclins for stabilization, activation, and downstream phosphorylation. In mammals, CDKs are primarily divided into subfamilies related to the cell cycle and those regulating transcription.

[0003] CDK12 / 13 (Cyclin-dependent kinase 12 / 13) are members of the cyclin-dependent kinase family (CDKs) of serine / threonine protein kinases. They form complexes with cyclin K to exert their biological functions. CDK12 / 13 contain 1490 and 1512 amino acids, respectively, and share 46% homology. The kinase domain, consisting of 300 amino acids, has a high homology of 92%. Compared to other transcriptional CDKs, CDK12 / 13 contain an additional arginine / serine-rich (RS) motif at the N-terminus. RS motifs are commonly found in proteins involved in pre-mRNA splicing. Proline-rich motifs (PRIM) motifs are also found at the N- and C-termini, potentially serving as binding sites for proteins containing SH3, WW, or actin-binding protein (profilin) ​​domains. CDK12 / 13 form a complex with cyclin K by phosphorylating the C-terminal domain (CTD) of RNA polymerase II (RNA Pol II). The CTD consists of a highly repetitive sequence consisting of seven amino acids, YSPTSPS. In humans, the CTD consists of 52 repeats. CDK12 / 13 primarily phosphorylates the serine 2 at position 2, thereby regulating transcription and post-transcriptional mRNA processing. Genetic studies have shown that CDK12 promotes full-length gene transcription by inhibiting intronic polyadenylation sites. Many homologous recombination repair genes (such as BRCA1 / 2, ATM, ATR, FANCD2, and FANCI) contain more intronic polyadenylation sites, making their expression more sensitive to CDK12 depletion or inhibition. DNA double-strand breaks are highly toxic, and homologous recombination is the most accurate and error-free pathway for repairing double-strand breaks, making it crucial for DNA damage repair. Therefore, inhibiting CDK12 / CDK13 can induce DNA damage caused by homologous recombination repair deficiency (HRD) in cells, thereby killing cells. CDK12 / 13 inhibitors combined with other drugs have a synergistic effect to promote cancer cell killing, overcome drug resistance, and reduce the emergence of drug-resistant strains. In addition, CDK12 has synthetic lethal interactions with MYC, EWS / FLI fusion, and PARPi. CDK12 mutations have been reported in different proportions in castration-resistant prostate cancer, high-grade serous ovarian cancer, etc., most of which are nonsense mutations or inactivating mutations. CDK12 mutations are not a necessary condition for it to be a potential target for cancer treatment. Regulating the expression of DNA damage repair-related proteins and synthetic lethal interactions provide opportunities for it to be an anti-cancer target. Because CDK12 has great potential in disease treatment and can be used as a biomarker for cancer occurrence, it has attracted increasing interest from scientists in recent years. Summary of the Invention

[0004] Based on this, the present invention provides a class of novel CDK12 / 13 covalent inhibitors substituted with a ring structure or their pharmaceutical compositions as CDK12 / 13 inhibitors. These compounds can effectively and selectively inhibit CDK12 / 13 protein kinases and inhibit the proliferation, migration and invasion of various tumor cells.

[0005] In a first aspect, the present invention provides a CDK12 / 13 covalent inhibitor having a structure of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a prodrug molecule thereof:

[0006] Wherein, X and Y are independently selected from the following groups: N or CR5;

[0007] R5 is selected from the group consisting of hydrogen, halogen, cyano, hydroxy, amino, halomethyl, halomethoxy, haloethyl, haloethoxy, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, and C1-C6 alkyl-substituted amino;

[0008] Ring A is selected from:

[0009] Where s is 0, 1, 2 or 3;

[0010] F, G, H, I, K, L, P, Q, T, U, and Z are independently selected from the following groups: N or CR';

[0011] R' is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C1-C6 cycloalkyl, cycloalkoxy, (CH2)C1-C6 cycloalkyl;

[0012] J is selected from the group consisting of O, S, NH, NR";

[0013] R" is selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl;

[0014] R1 is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, C3-C8 cycloalkoxy;

[0015] W is selected from the group consisting of chemical bonds,

[0016] R2 is selected from the group consisting of H, -NHR6, -OR6, -CH(R8)R6;

[0017] R6 is selected from the group consisting of: -(C(R8)R7)R9, -(CH2) nR9; wherein n is selected from: 0, 1 or 2;

[0018] R7 and R8 are each independently selected from the group consisting of hydrogen, halogen, cyano, methyl, halomethyl, methoxy, halomethoxy, ethyl, haloethyl, ethoxy, haloethoxy, hydroxy, amino, and a 3-8 membered heterocycle containing 1, 2 or 3 heteroatoms; or R7 and R8 are joined together to form a 3-7 membered heterocycle containing 1, 2 or 3 heteroatoms through the carbon atoms to which they are joined; or R7 and R8, together with the carbon atoms to which they are joined, form a substituted or unsubstituted saturated C1-C8 monocyclic, condensed, spirocyclic or bridged ring;

[0019] R9 is selected from the group consisting of:

[0020] 1) C1-C8 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C3-C 10 Cycloalkyl, substituted or unsubstituted 3-8 membered aromatic ring or saturated ring containing O, S or N, 8-12 membered fused ring, spiro ring or bridged ring containing n heteroatoms, n is selected from: 1, 2 or 3, heteroatoms are selected from: O, N, S;

[0021] 2)

[0022] A, B, C, D, and E are independently selected from: CH, N, or CR 10 ;

[0023] R 10 Selected from the group consisting of halogen, cyano, hydroxy, amino, nitro, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, C3-C8 cycloalkyl;

[0024] A', B', C', D', E' are independently selected from CH, N, NH, S, O, NR 10' or CR 10”

[0025] R 10' 、R 10” Each independently selected from: C1-C5 alkyl, C1-C5 cycloalkyl;

[0026] V is N or CR3;

[0027] Each R3 is independently selected from the following group: H, halogen, cyano, hydroxy, amino, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, halogenated C1-C3 alkoxy, C3-C8 cycloalkyl, -(CH2) m R 11 、-NH(CH2) m R 11 、-NR 14 (CH2)m R 11 、-O(CH2) m R 11 ; a substituted or unsubstituted 3-8 membered heterocyclic ring having 1, 2 or 3 heteroatoms; an 8-12 membered fused, spiro or bridged ring containing 1, 2 or 3 heteroatoms; and at least one R3 is not H;

[0028] R 11 Selected from the following group: C1-C6 alkyl or NR 12 R 13 ; Among them, R 12 、R 13 are independently selected from: H, C1-C8 alkyl, -(CH2) m NR 14 R 15 、-(CH2) n CR 14 R 15 R 16 , or R 12 、R 13 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted 4-12 membered monocyclic, condensed, spirocyclic or bridged ring containing 0-3 heteroatoms;

[0029] R 14 、R 15 、R 16 Each independently selected from the following group: H, C1-C8 alkyl, or R 14 、R 15 Together with the nitrogen atom or carbon atom to which they are attached, they form a substituted or unsubstituted 4-12 membered monocyclic, condensed, spirocyclic or bridged ring containing 0-3 heteroatoms;

[0030] m and n are independently selected from the integers of 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0031] R4 is selected from the following group: H,

[0032] R 17 Selected from the group consisting of hydrogen, trifluoromethyl,

[0033] R 18 Selected from the group consisting of hydrogen, fluorine or methyl;

[0034] Unless otherwise specified, the term "substituted" refers to the replacement of one or more hydrogen atoms on a group by a substituent selected from the group consisting of halogen, oxo, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C1-C6 acyl, unsubstituted or halogenated C1-C6 amide, unsubstituted or halogenated C1-C6 alkylamino, unsubstituted or halogenated C1-C6 alkyl-hydroxy, unsubstituted or halogenated C3-C6 alkyl, or a 4-8 membered heterocyclyl which is unsubstituted, halogenated or substituted by C1-C4 alkyl; any of the heteroatoms being selected from the group consisting of O, N, and S.

[0035] In another preferred embodiment, X and Y are independently selected from: N or CR5;

[0036] R5 is selected from the group consisting of hydrogen, halogen, cyano, halomethyl, and halomethoxy.

[0037] In another preferred embodiment, ring A is selected from the following group:

[0038] Where s is 0, 1, 2 or 3;

[0039] F, G, H, I, K, L, P, Q, U, and Z are independently selected from the following groups: N or CR';

[0040] R' is selected from the group consisting of H, cyano, halogen,

[0041] J is selected from the group consisting of O, S, NH, NR";

[0042] R" is selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl;

[0043] R1 is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy;

[0044] In another preferred embodiment, W is selected from:

[0045] In another preferred embodiment, R2 is selected from the following group: -NHR6;

[0046] R6 is selected from the group consisting of: -(C(R8)R7)R9;

[0047] R7 and R8 are each independently selected from the group consisting of hydrogen, halogen, cyano, methyl, halomethyl, methoxy, halomethoxy, ethyl, or R7 and R8 together with the carbon atoms to which they are attached form a saturated C1-C6 monocyclic, spirocyclic or bridged ring;

[0048] R9 is independently selected from the following group: phenyl, halophenyl, cyano-substituted phenyl, alkoxy-substituted phenyl, pyridyl, pyrimidinyl, isopropyl, tert-butyl, trifluoromethyl, difluoromethyl, cyano, substituted or unsubstituted pyrrolyl, N-methylpyrrolyl, N-methylimidazolyl, N-methylpyrazolyl, imidazolyl, substituted or unsubstituted furanyl, substituted or unsubstituted thienyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted isoxazolyl, substituted or unsubstituted oxazolyl, halo-substituted C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkyl, C3~C7 cycloalkyl, C3~C7 epoxyalkyl.

[0049] In another preferred embodiment, each R3 is independently selected from the following group: H, halogen, cyano, hydroxyl, amino, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, halogenated C1-C3 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy,

[0050] In another preferred embodiment, R4 is selected from:

[0051] H.

[0052] In another preferred embodiment, it has a structure shown in formula (II):

[0053] In another preferred embodiment, the compound is selected from the following compounds:

[0054] The second aspect of the present invention provides a use of the CDK12 / 13 covalent inhibitor or its pharmaceutical composition according to the first aspect of the present invention, or its pharmaceutically acceptable salt, or its stereoisomer or its prodrug molecule in the preparation of a drug for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinase.

[0055] In another preferred embodiment, the disease mediated by CDK12 / 13 serine / threonine protein kinase is selected from the following group: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing 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, and glioma.

[0056] The third aspect of the present invention provides a pharmaceutical composition for preventing and / or treating tumors, which comprises an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient comprises a novel CDK12 / 13 covalent inhibitor substituted with a paracyclic structure as described in the first aspect of the present invention, or a pharmaceutical composition thereof, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof.

[0057] 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 described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0058] In some of the embodiments, a class of novel CDK12 / 13 covalent inhibitors substituted with a cyclic structure or their pharmaceutical compositions in the preparation of CDK12 / 13 inhibitors is provided.

[0059] Another object of the present invention is to provide a use of the above-mentioned CDK12 / 13 covalent inhibitor or its pharmaceutical composition in the preparation of a method for preventing and / or treating diseases caused by CDK12 / 13 inhibitors.

[0060] In some embodiments, the disease mediated by the CDK12 / 13 serine / threonine protein kinase is preferably any one of: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing 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, and glioma.

[0061] Another object of the present invention is to provide a pharmaceutical composition for preventing and / or treating tumors, comprising an active ingredient and a pharmaceutically acceptable excipient, wherein the active ingredient comprises the above-mentioned trans-1,4-cyclohexanediamine compound or a pharmaceutically acceptable salt or stereoisomer thereof or a prodrug molecule thereof.

[0062] the term

[0063] The experimental methods in the following examples of the present invention, where no specific conditions are specified, are generally carried out under conventional conditions or conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0064] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0065] The terms "comprise," "comprising," and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, product, or device comprising a series of steps is not limited to the listed steps or modules but may optionally include steps not listed, or other steps inherent to the process, method, product, or device.

[0066] In this application, "plurality" refers to two or more. "And / or" describes the relationship between related objects, indicating that three possible relationships exist. For example, "A and / or B" can mean: A exists alone, A and B exist simultaneously, or B exists alone. The character " / " generally indicates that the related objects are in an "or" relationship.

[0067] In the compounds of the present invention, when any variable (such as R 10 、R 11 If a substituent (e.g., ) occurs more than once in any component, its definition at each occurrence is independent of its definition at every other occurrence. Likewise, combinations of substituents and variables are permissible so long as such combinations result in a stable compound. A line drawn from a substituent into the ring system indicates that the indicated bond may be attached to any substitutable ring atom. If the ring system is polycyclic, this means that such bonds may be attached only to any suitable carbon atom in an adjacent ring. It will be understood that one of ordinary skill in the art can select substituents and substitution patterns in the compounds of the present invention to provide compounds that are chemically stable and readily synthesized from readily available starting materials using techniques in the art and the methods set forth below. If a substituent is itself substituted with more than one group, it will be understood that these groups may be on the same carbon atom or on different carbon atoms so long as the structure is stable.

[0068] As used herein, the term "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms. For example, the definition of "C1-C8" in "C1-C8 alkyl" includes groups having 1, 2, 3, 4, 5, or 8 carbon atoms in a straight or branched arrangement. The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having a specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and the like.

[0069] The term "alkoxy" as used herein represents an alkyl-oxy group wherein alkyl is as defined above.

[0070] As used herein, the term "cycloalkyl" refers to a cyclic aliphatic alkane having a specified number of carbon atoms, wherein the ring backbone is entirely carbon atoms. In some preferred embodiments, the carbocyclic ring may be saturated or partially unsaturated, but is not aromatic. For example, a C3-C8 cycloalkyl refers to a cyclic alkyl group having 3, 4, 5, 6, 7, or 8 carbon atoms as the ring backbone, such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, or similar structures.

[0071] The term "aryl" as used herein refers to an aromatic cyclic group having a specific number of carbon atoms, for example, an aromatic group having 6, 7, 8, 9 or 10 carbon atoms, including monocyclic or bicyclic aromatic groups, such as phenyl, naphthyl, or the like.

[0072] The term "heterocyclyl" refers to a saturated or unsaturated ring substituent having a specified number of ring atoms, but is not aromatic. For example, a 3-10 membered heterocyclyl refers to a saturated or unsaturated ring substituent having 3, 4, 5, 6, 7, 8, 9, or 10 members, wherein the ring backbone includes at least one heteroatom selected from O, S, or N, such as pyridyl, thienyl, piperidinyl, morpholinyl, dihydropiperidinyl, thiomorpholinyl, piperidinyl, piperazinyl, tetrahydropyranyl, dihydropyranyl, pyrrolinyl, tetrahydrothienyl, tetrahydrofuranyl, oxetanyl, thietanyl, azetidinyl, or similar groups, preferably a 4-9 membered heterocyclyl.

[0073] As used herein, the term "heteroaryl" refers to an aromatic cyclic group having a specified number of ring atoms, wherein at least one ring atom is a heteroatom selected from O, S or N, for example, a heteroaryl group having 5, 6, 7, 8, 9 or 10 ring atoms, including monocyclic or bicyclic heteroaryl groups, such as pyridine, pyrimidine, pyridazine, tetrazine, triazine, pyrrole, thiophene, furan, tetrazole, triazole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, oxadiazole, thiadiazole, naphthalene, indole, indazole, quinoline, isoquinoline, benzofuran, benzothiophene, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, benzisoxazole, benzotriazole, or the like.

[0074] As will be understood by those skilled in the art, "halogen" as used herein is meant to include chlorine, fluorine, bromine, and iodine.

[0075] The present invention includes the free form of the compound of formula (I), as well as its pharmaceutically acceptable salts, stereoisomers, and prodrug molecules. The term "free form" refers to the compound in a non-salt form. Pharmaceutically acceptable salts included include not only the exemplary salts of the specific compounds described herein, but also all typical pharmaceutically acceptable salts of the free form of the compound of formula (I). The free form of a specific salt of the compound can be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a suitable dilute aqueous base solution, such as a dilute aqueous solution of NaOH, a dilute aqueous solution of potassium carbonate, a dilute aqueous ammonia, and a dilute aqueous solution of sodium bicarbonate. The free form differs somewhat from its respective salt form in certain physical properties, such as solubility in polar solvents, but for the purposes of the invention, such acid and base salts are equivalent to their respective free forms in other pharmaceutical aspects.

[0076] Compounds and pharmaceutically acceptable salts

[0077] Disclosed herein are compounds of formula (I), including compounds of formula (II) (the structures of which are disclosed herein), and pharmaceutically acceptable salts thereof.

[0078] Pharmaceutically acceptable salts of the present invention can be synthesized from compounds of the present invention containing a basic or acidic moiety by conventional chemical methods. Typically, salts of basic compounds are prepared by ion exchange chromatography or by reacting a free base with a stoichiometric amount or an excess of an inorganic or organic acid in the desired salt form in a suitable solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reaction with a suitable inorganic or organic base.

[0079] Therefore, the pharmaceutically acceptable salts of the compounds of this invention include conventional non-toxic salts of the compounds of this invention formed by reacting an alkaline compound of this invention with an inorganic or organic acid. For example, conventional non-toxic salts include salts prepared from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and salts prepared 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, isethionic acid, trifluoroacetic ....

[0080] If the compound of the present invention is acidic, suitable "pharmaceutically acceptable salts" refer to salts prepared from 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, ferric salts, ferrous salts, lithium salts, magnesium salts, manganic salts, manganous salts, potassium salts, sodium salts, zinc salts, and the like. Ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic 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, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, guaiac, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

[0081] Berg et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977: 66: 1-19, describe in more detail the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts.

[0082] The compounds disclosed herein include at least one asymmetric or chiral center and can therefore exist as stereoisomers. Each chiral center is "R" or "S," depending on the configuration of substituents around the chiral carbon atom. The terms "R" and "S" as used herein are as defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem. 1976, 45: 13-30. Various stereoisomers and mixtures thereof are specifically included within the scope of this disclosure, including enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. Single stereoisomers of a compound can be synthesized from commercially available starting materials containing an asymmetric or chiral center, or by preparing a racemic mixture followed by resolution methods well known to those of ordinary skill in the art. Examples of these resolution methods are as follows: (1) attaching a mixture of enantiomers to a chiral auxiliary, separating the resulting diastereomeric mixture by recrystallization or chromatography, and optionally liberating the optically pure product from the auxiliary, as described in Furniss, Hannaford, Smith, and Tatchell, "Vogel's Textbook of Practical Organic Chemistry," 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England; (2) direct separation of the mixture of optical enantiomers on a chiral chromatographic column; and (3) fractional recrystallization.

[0083] The compounds disclosed herein may exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure.

[0084] Compounds of the present disclosure can also be prodrug forms. As used herein, the term "prodrug" refers to a compound that produces an active compound when metabolized (e.g., in vivo or in vitro). In some embodiments, a prodrug can be inactive, or have an activity lower than a free drug, but can provide favorable processing, administration or metabolic characteristics. Exemplary prodrug moieties of the present invention can be connected to a free drug by a hydroxyl group, an amino group, a phosphate ester or a thiophosphate backbone of a nucleotide, and can include esters, carbamates, carbonyls, thioesters, amides, isocyanates, ureas, thioureas or other physiologically acceptable metabolically unstable parts. In some embodiments, a prodrug is activated by enzymatic hydrolysis.

[0085] The present disclosure also includes isotopically labeled compounds, which are the same as those listed in Formula (I) and Formula (II), but one or more atoms are replaced by an atom having a mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the present disclosure 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 respectively. With heavier isotopes (such as deuterium, i.e. 2 H) substitution may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in certain circumstances. The compounds may be combined with positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies for determining receptor distribution. Suitable positron-emitting isotopes that may be incorporated into the compounds of formula (I) and (II) are 11 C. 13 N. 15 O and 18 F. Isotopically labeled compounds of formula (I), (II) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically labeled reagent in place of a non-isotopically labeled reagent.

[0086] The compounds disclosed herein can exist in solvated and unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and the disclosure is intended to encompass both 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 a crystalline form.

[0087] Pharmaceutical compositions and methods of administration

[0088] Since the compounds of the present disclosure are CDK12 / 13 inhibitors, the compounds of the present disclosure and pharmaceutically acceptable salts thereof, as well as other compound forms disclosed herein, can be included in pharmaceutical compositions useful for treating, preventing, and alleviating diseases related to the activity of CDK12 / 13.

[0089] The pharmaceutical composition of the present invention comprises 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" refers to an amount sufficient to induce a desired biological response (e.g., to treat a condition). "Safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition comprises 1 to 3000 (active dose range is 3 to 30 mg / kg) mg of the compound of the present invention / agent, more preferably 10 to 2000 mg of the compound of the present invention / agent. Preferably, "one dose" is a capsule or tablet.

[0090] "Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel substances suitable for human use, which must be of sufficient purity and sufficiently low toxicity. "Compatibility" in this context means that the components of the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier parts include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose 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 and olive oil), polyols (such as propylene glycol, glycerol, mannitol and sorbitol), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives and pyrogen-free water.

[0091] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), or topical administration.

[0092] Solid preparations for oral administration include capsules, tablets, pills, powders and granules. In these solid preparations, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or fillers, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, 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) retarding solvents, such as paraffin; (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 the case of capsules, tablets and pills, the formulation may also comprise buffering agents.

[0093] Solid dosage forms, such as tablets, dragees, capsules, pills, and granules, can be prepared with coating and shell materials, such as enteric coatings and other materials well known in the art. They may contain opacifiers, and the active compound or compounds in such compositions may be released in a delayed manner in a portion of the digestive tract. Examples of useful embedding ingredients are polymers and waxes. If desired, the active compound may also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0094] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0095] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0096] Suspensions, in addition to the active compounds, may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanolate and agar, or mixtures thereof.

[0097] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0098] Dosage forms for topical administration of the disclosed compounds include ointments, powders, patches, sprays and inhalants.The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers or propellants, if required.

[0099] The compounds of the present disclosure may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0100] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, at a pharmaceutically effective dose. For a 60 kg human, the daily dose is typically 1-2000 mg, preferably 6-600 mg. Of course, the specific dosage should also be determined taking into account factors such as the route of administration and the patient's health status, all of which are within the skill of a skilled physician.

[0101] Therapeutic uses and methods

[0102] As described above, the compounds of the present disclosure are CDK12 / 13 inhibitors, and thus the compounds or compositions comprising the compounds can be used to treat, prevent, and alleviate diseases associated with the activity or abnormal expression of CDK12 / 13. In some embodiments, disclosed herein is the use of the compounds disclosed herein in the preparation of medicaments for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinases. In some embodiments, disclosed herein are compounds disclosed herein for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinases. In some embodiments, disclosed herein is a method for treating a disease mediated by CDK12 / 13 serine / threonine protein kinases in a subject in need thereof, comprising administering to the subject an effective amount of a compound disclosed herein. In some embodiments, diseases mediated by CDK12 / 13 serine / threonine protein kinases include prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing 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 cancer, colon cancer, rectal cancer, and glioma.

[0103] When used for purposes and methods disclosed herein, disclosed compounds and compositions can be used in combination with other known therapies. As used herein, "combined" administration refers to giving two (or more) different treatments to a subject during the course of the subject suffering from the disease, for example, after the subject is diagnosed, the disease is cured or eliminated, or two or more treatments are given before treatment is stopped for other reasons. In some embodiments, when the second treatment begins, one treatment is still ongoing, so there is overlap in terms of treatment. This is referred to as "simultaneously" or "concurrently" in this article in some cases. In other embodiments, one treatment ends before another treatment begins. In some embodiments of either case, combined therapy is more effective.

[0104] For example, the second treatment shows a better effect, such as the same effect can be seen with less second treatment than when the second treatment is applied in the absence of the second treatment, or the second treatment alleviates the symptoms to a greater extent, or a similar situation is found in the first treatment. In some embodiments, such a combined treatment makes the improvement of the symptoms or other parameters associated with the disease greater than the improvement observed when one treatment is performed in the absence of another treatment. The effects of the two treatments can be partially cumulative, completely cumulative, or greater than cumulative. The treatment can make the effect of the first treatment still detectable when the second treatment is performed.

[0105] The compounds or compositions disclosed herein and at least one additional therapeutic agent can be administered simultaneously, in the same or different compositions, or sequentially. For sequential administration, the compounds as described herein can be administered first, followed by the additional agent, or the order of administration can be reversed.

[0106] In some embodiments, the compounds described herein are administered in combination with other therapeutic treatments, including surgery, radiation, transplantation (e.g., stem cell transplantation, bone marrow transplantation), chemotherapy, immunotherapy, cryotherapy, and / or thermotherapy. Such combination therapies can allow for lower doses of the administered agent and / or other agents, thereby avoiding possible toxicities or complications associated with various therapies.

[0107] 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.

[0108] Compared with the prior art, the present invention has the following beneficial effects:

[0109] The novel CDK12 / 13 covalent inhibitor with a cyclic structure substitution provided by the present invention or its pharmaceutical composition can effectively inhibit CDK12 protein kinase and can be used to prepare drugs for preventing or treating diseases mediated by CDK12 protein kinase, such as prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing 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., and at the same time has strong inhibitory activity, kinase selectivity and good pharmacokinetic characteristics.

[0110] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0111] Example 1: Compound ZLC-7-42

[0112] Step 1: Synthesis of intermediate F-2-3

[0113] In a 100 mL round-bottom flask, 4-dimethylaminopiperidine (2.63 g, 20.5 mmol) and cesium carbonate (8.0 g, 24.6 mmol) were added sequentially to a system of 2-fluoro-4-bromonitrobenzene (4.5 g, 20.5 mmol) in acetonitrile (10 mL). After completion, the temperature was raised to 60°C and the reaction was allowed to proceed overnight. After completion of the reaction as detected by TLC, the mixture was brought to room temperature, filtered, and the filtrate was concentrated under reduced pressure. 100 mL of water was added, and the mixture was extracted with CH2Cl2 / MeOH (10:1). The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the target product F-2-3 (yellow solid, 6.37 g, yield 95%). 1 H NMR(500MHz,DMSO-d6)δ7.98(d,J=2.5Hz,1H),7.69(dd,J=8.9,2.5Hz,1H),7.23(d,J=8.9Hz,1H),3.19 –3.16(m,1H),2.81–2.75(m,2H),2.22–2.17(m,7H),1.79–1.76(m,2H),1.49–1.40(m,2H).MS(ESI)for C 13 H 18 BrN3O2[M+H] + ,calcd:327.1,found:328.2.

[0114] Step 2: Synthesis of intermediate F-2-6

[0115] In a three-necked round-bottom flask, intermediate F-2-3 (1.5 g, 4.9 mmol), trans-(4-aminocyclohexyl)carbamic acid tert-butyl ester (3.2 g, 14.7 mmol), CuI (0.1 g, 0.5 mmol), D-proline (58 mg, 0.5 mmol), and K3PO4 (2.1 g, 9.8 mmol) were added in sequence, and the mixture was purged with argon three times. DMSO (22 mL) was added, and the temperature was raised to 100°C. After reacting overnight, the mixture was brought to room temperature and filtered through celite. The filtrate was concentrated under reduced pressure, 100 mL of water was added, and the mixture was extracted with CH2Cl2 / MeOH (10:1) system. The mixture was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the target product F-2-6 (gray solid, 1.6 g, yield 71%). 1 H NMR(500MHz,DMSO-d6)δ7.16(d,J=8.9Hz,1H),6.83(d,J=2.7Hz,1H),6.80–6.7 5(m,2H),5.78(d,J=8.1Hz,1H),3.24–3.17(m,1H),3.11–3.06(m,1H),2.97(d,J =11.4Hz,2H),2.64(t,J=10.7Hz,2H),2.18(m,7H),1.93(d,J=12.0Hz,2H),1.7 6(m,4H),1.42(m,2H),1.38(s,9H),1.31–1.21(m,2H),1.13(m,2H).MS(ESI)for C 24 H 39 N5O4[M+H] + ,calcd:462.3,found:462.4.

[0116] Step 3: Synthesis of intermediate F-2-8

[0117] In a round-bottom flask, CF3COOH (1 mL) was added dropwise to a CH2Cl2 (2 mL) solution of the intermediate F-2-6 (0.8 g, 1.74 mmol) and stirred at room temperature. After the reaction was completed, the mixture was concentrated under reduced pressure, neutralized with saturated sodium carbonate and adjusted to alkaline. The mixture was extracted with CH2Cl2 / MeOH (10:1), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the target product F-2-8 (dark gray oily liquid, 0.61 g, yield 97%). 1H NMR(500MHz,DMSO-d6)δ7.16(d,J=8.9Hz,1H),6.82(d,J=2.6Hz,1H),6.76( dd,J=8.9,2.7Hz,1H),5.75(d,J=7.8Hz,1H),3.20–3.17(m,1H),3.12–3.05( m,1H),2.98–2.95(m,2H),2.64(t,J=10.9Hz,2H),2.18–2.10(m,7H),1.90–( m,2H),1.75–1.74(m,4H),1.44–1.37(m,2H),1.17–1.07(m,4H).MS(ESI)for C 19 H 31 N5O2[M+H] + ,calcd:362.3,found:362.4.

[0118] Step 4: Synthesis of intermediate F-2-27

[0119] In a 25 mL round-bottom flask, 2-chloroquinazoline (118 mg, 0.72 mmol) and cesium carbonate (255 mg, 0.78 mmol) were added to a DMF (2 mL) system of F-2-8 (236 mg, 0.65 mmol) in sequence. After completion, the temperature was raised to 60 ° C. After 6 h, the reaction was completed, filtered, and the filtrate was concentrated under reduced pressure. After purification by column chromatography, the target product F-2-27 (dark purple solid, 264 mg, yield 83%) was obtained. 1 H NMR(500MHz,DMSO-d6)δ9.09(s,1H),7.76(d,J=7.9Hz,1H),7.68–7.65(m,1H),7.45(d,J=7.2Hz,1H), 7.34(d,J=7.6Hz,1H),7.21–7.17(m,2H),6.88(d,J=2.5Hz,1H),6.81(dd,J=8.9,2.5Hz,1H),5.86(d, J=8.0Hz,1H),3.90–3.82(m,1H),3.23–3.14(m,1H),3.01–2.98(m,2H),2.68–2.64(m,2H),2.38–2.22 (m,7H),2.02–2.00(m,4H),1.80(d,J=12.0Hz,2H),1.49–1.41(m,4H),1.28–1.24(m,2H).MS(ESI)for C 27 H 35 N7O2[M+H] +,calcd:490.3,found:490.4.

[0120] Step 5: Synthesis of intermediate F-2-28

[0121] Under argon protection, triethylamine (28 mL) was added to a solution of benzylamine (10.7 g, 107 mmol) in anhydrous THF (400 mL) in a 1000 mL round-bottom three-necked flask. The mixture was then placed in an ice bath and p-nitrophenyl chloroformate (20.5 g, 102 mmol) in THF (100 mL) was slowly added dropwise. After completion, the mixture was stirred at room temperature. After completion of the reaction by TLC, the reaction was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and slurried in an ethyl acetate / petroleum ether system to obtain the target product (yellow-green solid, 16.6 g), which was directly used in the next step.

[0122] In a sealed tube, the above solid (881 mg, 3.24 mmol), intermediate F-2-27 (264 mg, 0.54 mmol), DIPEA, and DMF (2 mL) were added in sequence. The temperature was raised to 95 ° C. After 12 h, the mixture was transferred to room temperature and concentrated under reduced pressure. After purification by column chromatography, the target product F-2-28 (yellow solid, 225 mg, yield 67%) was obtained. 1 H NMR(500MHz,DMSO-d6)δ9.05(s,1H),7.74(d,J=8.1Hz,1H),7.65–7.59(m,2H),7.43–7.36(m,2H) ,7.34–7.31(m,1H),7.29–7.24(m,3H),7.18(m,4H),6.26–6.23(m,1H),4.29–4.24(m,1H),4.16( d,J=6.2Hz,2H),3.64–3.57(m,1H),3.28(m,2H),2.89–2.85(m,2H),2.36–2.19(m,7H),2.00–1.9 2(m,2H),1.88–1.81(m,4H),1.55–1.48(m,2H),1.45–1.38(m,2H),1.17–1.10(m,2H).MS(ESI)for C 35 H 42 N8O3[M+H] + ,calcd:623.3,found:623.7.

[0123] Step 6: Synthesis of intermediate F-2-29

[0124] In a 25 mL round-bottom flask, the intermediate F-2-28 in DMF (2 mL) was placed in an ice bath, and 4,4'-bipyridine (1 mg) and diboric acid (95 mg, 1.06 mmol) were added. After 30 min, the reaction was complete by TLC. The product was quenched with saturated sodium bicarbonate solution, filtered, and the filter cake was washed three times with distilled water and dried to obtain the target product F-2-29 (slightly yellow solid, 135 mg, yield 64%).

[0125] 1 H NMR(500MHz,DMSO-d6)δ9.05(s,1H),7.74(d,J=8.1Hz,1H),7.62(t,J=7.6Hz,1H),7.42(s,1H),7.29–7.23(m,3H),7. 19–7.16(m,4H),6.92(d,J=8.2Hz,1H),6.56–6.53(m,1H),6.37(dd,J=8.2,2.1Hz,1H),5.49(m,1H),4.88(s,2H),4.2 7–4.23(m,1H),4.17(d,J=5.9Hz,2H),3.65–3.56(m,1H),3.18(d,J=11.9Hz,2H),2.55(m,2H),2.44–2.33(m,7H),2.0 0–1.93(m,2H),1.90–1.87(m,2H),1.77–1.73(m,2H),1.67–1.61(m,2H),1.40(m,2H),1.20–1.16(m,2H).MS(ESI)for C 35 H 44 N8O[M+H] + ,calcd:593.4,found:593.6.

[0126] Step 7: Synthesis of compound ZLC-7-42

[0127] In a 25 mL round-bottom flask, DIPEA (0.1 mL) was added to a solution of intermediate F-2-29 (120 mg, 0.2 mmol) in anhydrous CH2Cl2 (2 mL) and the mixture was placed in an ice bath. After 15 min, acryloyl chloride (16 μL) was slowly added dropwise. The mixture was brought to room temperature and further acryloyl chloride was added until the reaction was complete. The mixture was quenched with saturated sodium bicarbonate solution and extracted with CH2Cl2 / MeOH (10:1). The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure. After purification by column chromatography, the target product ZLC-7-42 (white solid, 63 mg, 49% yield) was obtained. 1H NMR (600MHz, DMSO-d6) δ9.04(m,2H),7.96(s,1H),7.73(d,J=8.8Hz,1H),7.62(t,J=7.4Hz,1H),7.41(d,J=7.4Hz,1H),7 .29–7.15(m,8H),6.92(dd,J=8.3,2.3Hz,1H),6.75(dd,J=16.9,10.3Hz,1H),6.29(d,J=18.7Hz,1H),5.81–5.74(m,2H) ,4.30–4.26(m,1H),4.18(d,J=5.9Hz,2H),3.60–3.58(m,1H),3.10(d,J=11.5Hz,2H),2.67(t,J=11.1Hz,2H),2.38–2.3 3(m,7H),1.98–1.96(m,2H),1.90–1.88(m,2H),1.80–1.74(m,4H),1.45–1.38(m,2H),1.21–1.15(m,2H).HRMS(ESI)for C 38 H 46 N8O2[M+H] + ,calcd:647.3816,found:647.3816.

[0128] Example 2: Compound ZLC-7-55

[0129] The synthesis method is shown in Example 1.

[0130] 1H NMR (600MHz, DMSO-d6) δ9.28(s,1H),9.05(s,1H),8.03(s,1H),7.88(d,J=2.3Hz,1H),7.74(d,J=8.7Hz,1H),7.62(t,J=7.4Hz,1 H),7.40(m,1H),7.29-7.24(m,3H),7.19–7.16(m,4H),6.76(dd,J=16.9,10.3Hz,1H),6.31(dd,J=17.0,1.8Hz,1H),6.27–6.21(m ,1H),5.81(d,J=10.2Hz,1H),4.30–4.25(m,1H),4.18(d,J=5.8Hz,2H),3.61–3.54(m,3H),2.80–2.76(m,2H),2.65–2.54(m,1H) ,2.38(s,6H),2.02–1.94(m,2H),1.90(d,J=10.3Hz,2H),1.81–1.75(m,4H),1.46–1.40(m,2H),1.16–1.10(m,2H).HRMS(ESI)for C 37 H 45 N9O2[M+H] + ,calcd:648.3769,found:648.3770.

[0131] Example 3: Compound ZLC-7-81

[0132] The synthesis method is shown in Example 1.

[0133] 1H NMR (600MHz, DMSO-d6) δ9.26(s,1H),9.05(s,1H),8.02(s,1H),7.88(d,J=1.9Hz,1H),7.74(d,J=7.9Hz,1H),7.62(t,J=7 .5Hz,1H),7.41(d,J=7.6Hz,1H),7.27–7.24(m,3H),7.19–7.16(m,1H),7.12–7.10(m,3H),6.77(dd,J=16.9,10.3Hz,1H) ,6.52(s,1H),6.32(d,J=16.9Hz,1H),5.81(d,J=11.2Hz,1H),4.26–4.21(m,1H),3.61–3.55(m,3H),2.81–2.77(m,2H),2 .62–2.53(m,1H),2.41(s,6H),1.98–1.91(m,4H),1.80–1.78(m,4H),1.45–1.39(m,2H),1.15–1.05(m,6H).HRMS(ESI)for C 39 H 47 N9O2[M+H] + ,calcd:674.3922,found:674.3925.

[0134] Example 4: Compound ZLC-7-82

[0135] The synthesis method is shown in Example 1.

[0136] 1H NMR (600MHz, DMSO-d6) δ9.30(s,1H),9.04(s,1H),7.98(s,1H),7.87(d,J=2.3Hz,1H),7.77–7.70(m,1H),7.63–7.61(m,1H),7.41( d,J=7.7Hz,1H),7.31–7.22(m,5H),7.19–7.15(m,2H),6.76(dd,J=16.9,10.3Hz,1H),6.32(dd,J=17.0,1.8Hz,1H),5.90(d,J=7.4H z,1H),5.81(d,J=11.8Hz,1H),4.86(p,J=7.1Hz,1H),4.26–4.21(m,1H),3.60–3.58(m,3H),2.80(t,J=11.8Hz,2H),2.69–2.52(m, 1H),2.43(s,6H),1.99–1.92(m,4H),1.83–1.72(m,4H),1.46–1.37(m,2H),1.28(d,J=7.1Hz,3H),1.17–1.05(m,2H).HRMS(ESI)for C 38 H 47 N9O2[M+H] + ,calcd:662.3923,found:662.3925.

[0137] Example 5: Compound ZLC-7-83

[0138] The synthesis method is shown in Example 1.

[0139] 1H NMR (600MHz, DMSO-d6) δ9.30(s,1H),9.04(s,1H),7.98(s,1H),7.87(s,1H),7.73(d,J=7.9Hz,1H),7.62(t,J=7.2Hz,1H),7.41 (d,J=6.8Hz,1H),7.31–7.22(m,5H),7.17(t,J=7.1Hz,2H),6.76(dd,J=16.8,10.3Hz,1H),6.33–6.30(m,1H),5.90(d,J=6.9Hz ,1H),5.81(d,J=10.5Hz,1H),4.86(p,J=7.1Hz,1H),4.25–4.21(m,1H),3.59(m,3H),2.80(t,J=12.0Hz,2H),2.72–2.57(m,1H) ,2.43(s,6H),1.99–1.91(m,4H),1.85–1.70(m,4H),1.47–1.36(m,2H),1.27(d,J=7.0Hz,3H),1.17–1.05(m,2H).HRMS(ESI)for C 38 H 47 N9O2[M+H] + ,calcd:662.3922,found:662.3925.

[0140] Example 6: Compound ZLC-8-13

[0141] The synthesis method is shown in Example 1.

[0142] 1H NMR(600MHz,DMSO-d6)δ10.54(s,1H),9.25(s,1H),8.96(s,1H),7.99(s,1H),7.95–7.86(m,2H),7.78(s,1H),7.60(s,1H ),7.35–7.25(m,3H),7.11–7.08(m,2H),5.97(s,1H),5.94(s,1H),5.62(s,1H),4.86(p,J=7.1Hz,1H),4.26–4.21(m,1H), 3.67–3.62(m,3H),3.36–3.31(m,1H),2.97–2.81(m,2H),2.77(d,J=4.9Hz,6H),2.48–2.46(m,1H),2.17(d,J=11.0Hz,2H) ,2.05(s,3H),1.98(s,2H),1.85–1.75(m,4H),1.52–1.44(m,2H),1.27(d,J=7.1Hz,3H),1.18–1.11(m,2H).HRMS(ESI)for C 39 H 48 FN9O2[M+H] + ,calcd:694.3984,found:694.3988.

[0143] Example 7: Compound ZLC-8-23

[0144] The synthesis method is shown in Example 1.

[0145] 1H NMR (600MHz, DMSO-d6) δ9.27(s,1H),9.04(s,1H),7.94(s,1H),7.87(d,J=2.2Hz,1H),7.73(d,J=7.2Hz,1H),7.62(t,J=7.4Hz,1H),7.41(d,J=7.4H z,1H),7.37–7.32(m,1H),7.24(d,J=7.9Hz,1H),7.18–7.16(m,1H),7.14– 7.10(m,1H),7.03–7.00(m,1H),6.76(dd,J=16.9,10.2Hz,1H),6.31(d,J= 15.2Hz,1H),6.03(d,J=7.2Hz,1H),5.81(d,J=11.8Hz,1H),5.07(p,J=7.1 Hz,1H),4.23–4.17(m,1H),3.59–3.55(m,3H),2.78(t,J=12.3Hz,2H),2.2 9–2.17(m,7H),1.99–1.96(m,2H),1.86–1.79(m,3H),1.70–1.69(m,3H),1 .46–1.35(m,2H),1.24(d,J=7.0Hz,3H),1.17–1.03(m,2H).HRMS(ESI)for C 38 H 45 F2N9O2[M+H] + ,calcd:698.3733,found:698.3737.

[0146] Example 8: Compound ZLC-8-25

[0147] The synthesis method is shown in Example 1.

[0148] 1H NMR(600MHz,DMSO-d6)δ9.28(s,1H),9.04(s,1H),7.94(s,1H),7.88(d,J=2 .3Hz,1H),7.73(d,J=8.7Hz,1H),7.62(t,J=7.5Hz,1H),7.41(dd,J=8.7,6.3 Hz,2H),7.33(dd,J=8.8,2.7Hz,1H),7.24(d,J=7.9Hz,1H),7.20–7.15(m,2H ),6.77(dd,J=16.9,10.3Hz,1H),6.32(dd,J=17.0,1.8Hz,1H),6.18(d,J=7. 3Hz,1H),5.81(dd,J=10.3,1.6Hz,1H),5.13(p,J=7.1Hz,1H),4.21–4.16(m, 1H),3.61–3.53(m,3H),2.78(td,J=12.0,3.8Hz,2H),2.27–2.21(m,7H),2.0 0–1.90(m,2H),1.85(d,J=11.2Hz,2H),1.83–1.78(m,1H),1.74–1.65(m,3H) ,1.44–1.34(m,2H),1.22(d,J=7.1Hz,3H),1.17–1.02(m,2H).HRMS(ESI)for C 38 H 45 ClFN9O2[M+H] + ,calcd:714.3438,found:714.3442.

[0149] Example 9: Compound ZLC-8-53

[0150] The synthesis method is shown in Example 1.

[0151] 1H NMR (600MHz, DMSO-d6) δ9.29(s,1H),9.04(s,1H),7.95(dd,J=17.1,2.0Hz,2H),7.73(d,J=7.8Hz,1H),7.62(t,J=7.6Hz,1H),7.39(d,J=8.4Hz, 1H),7.29–7.25(m,3H),7.17(t,J=7.4Hz,1H),7.12–7.06(m,2H),5.99(d,J=7.5Hz,1H),5.80(dd,J=48.6,3.8Hz,1H),5.53(dd,J=15.8,3.8Hz, 1H),4.86(p,J=7.1Hz,1H),4.25–4.21(m,1H),3.56(d,J=12.1Hz,3H),2 .82(t,J=11.7Hz,2H),2.64–2.53(m,1H),2.38(s,6H),2.05–1.89(m,4H ),1.82(d,J=12.1Hz,1H),1.74(d,J=12.0Hz,1H),1.63–1.58(m,2H),1.46–1.37(m,2H),1.27(d,J=7.1Hz,3H),1.16–1.04(m,2H).HRMS(ESI)for C 38 H 45 F2N9O2[M+H] + ,calcd:698.3737,found:698.3737.

[0152] Example 10: Compound YJZ7001

[0153] The synthesis method is shown in Example 1.

[0154] 1H NMR (600MHz, DMSO-d6) δ9.05(s,1H),9.04(s,1H),7.95(s,1H),7.74(d,J=7.7Hz,1H),7.63(t,J=7.7Hz,1H),7.42(d,J=8.5Hz,1H),7. 30–7.22(m,4H),7.21–7.15(m,4H),6.95(dd,J=8.4,2.4Hz,1H),6.67(dd,J=16.9,10.3Hz,1H),6.29(dd,J=17.0,1.7Hz,1H),5.80(dd, J=10.1,1.8Hz,2H),4.32–4.26(m,1H),4.18(d,J=6.0Hz,2H),3.60(d,J=10.4Hz,2H),2.96–2.79(m,4H),2.48(t,J=10.3Hz,2H),2.37( s,1H),2.26(s,3H),1.99–1.92(m,2H),1.80(d,J=11.9Hz,2H),1.43(q,J=11.9,11.3Hz,2H),1.21–1.16(m,2H),1.02(d,J=6.2Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ163.63,162.48,157.03,142.82,141.72,134.41 ,133.80,132.99,132.60,128.50(4C),128.29,127.58,127.50,127.11(4 C),126.69,124.52,122.16,120.91,119.96,58.90,57.62,53.85,52.45, 51.83,49.03,43.93,42.63,40.52,31.75,30.85,17.19.HRMS(ESI)calcd for C 37 H 44 N8O2[M+H] + ,633.3660;found,633.3649.

[0155] Example 11: Compound YJZ1125

[0156] The synthesis method is similar to that of Example 1.

[0157] 1H NMR (400MHz, DMSO-d6) δ9.02(s,1H),8.72(s,1H),7.90(d,J=16.7Hz,2H),7.35–7.12(m,7H),6.94(d,J=8.4Hz,1H ),6.66(dd,J=17.0,10.2Hz,1H),6.27(d,J=16.9Hz,1H),5.84–5.70(m,2H),4.26(t,J=11.8Hz,1H),4.17(d,J=6.0 Hz,2H),3.74(s,3H),3.63–3.47(m,1H),2.97–2.76(m,4H),2.47(t,J=10.4Hz,2H),2.40–2.32(m,1H),2.26(s,3H) ,2.04–1.89(m,2H),1.79(d,J=10.5Hz,2H),1.41(q,J=12.6Hz,2H),1.17(q,J=13.3Hz,2H),1.01(d,J=6.0Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ163.64,160.66,157.04,154.22,142.84,141.70,133.80,133.30,132.96,132.57,128.50(4C),127.52,127.10(3C ),126.70,124.55,120.93,58.86,57.61,55.38,51.81,49.06,43.92,42.62,40.50,33.19,31.70(2C),30.82(2C),17.17.HRMS(ESI)calcd for C 35 H 44 N 10 O2[M+H] + ,637.3721;found,637.3738.

[0158] Example 12: Compound YJZ7027

[0159] The synthesis method is similar to that of Example 1.

[0160] 1H NMR (400MHz, DMSO-d6) δ9.02(s,1H),8.73(s,1H),7.92(s,1H),7.89(s,1H),7.32–7.24(m,3H),7.23(d,J=8.3Hz,1H),7.21–7.15(m 3H),6.94(d,J=8.4Hz,1H),6.66(dd,J=16.9,10.3Hz,1H),6.27(dd,J=16.9,1.8Hz,1H),5.83–5.7 7(m,1H),5.75(s,1H),4.27(t,J=12.4Hz,1H),4.21–4.11(m,4H),3.51(s,1H),2.97–2.76(m,4H), 2.46(d,J=10.7Hz,1H),2.41–2.31(d,J=9.9Hz,1H),2.25(s,3H),2.04–1.92(m,2H),1.79(d,J=11 .9Hz,2H),1.41(q,J=13.1Hz,2H),1.31(t,J=7.2Hz,3H),1.22–1.10(m,2H),1.01(d,J=6.1Hz,3H). 13 CNMR(151MHz,DMSO-d6)δ163.61,160.54,157.03,154.24,142.83,141.72 ,133.83,133.32,132.98,132.59,130.12,128.50(4C),127.50,127.10(3C ),126.69,124.50,120.91,58.87,57.61,51.82,49.04,43.92,42.63,40. 91,40.52,31.70,30.81(2C),29.50,27.02,17.16,14.95.HRMS(ESI)calcd for C 36 H 46 N 10 O2[M+H] + ,651.3878,found,651.3869.

[0161] Example 13: Compound YJZ7042

[0162] The synthesis method is similar to that of Example 1.

[0163] 1H NMR (400MHz, DMSO-d6) δ9.02(s,1H),8.73(s,1H),7.91(d,J=12.7Hz,2H),7.32–7.13(m,7H),6.94(d,J=8.3H z,1H),6.66(dd,J=16.9,10.3Hz,1H),6.27(dd,J=17.0,1.9Hz,1H),5.83–5.71(m,2H),4.30–4.21(m,1H),4.1 8(d,J=6.0Hz,2H),4.09(t,J=6.8Hz,2H),3.51(s,1H),2.97–2.75(m,4H),2.45(d,J=10.8Hz,2H),2.36(s,1H) ,1.97(s,2H),1.77(q,J=7.2Hz,4H),1.40(q,J=12.3Hz,2H),1.18(s,2H),1.01(d,J=6.2Hz,3H),0.77(s,3H). 13 CNMR(151MHz,DMSO-d6)δ163.61,160.57,157.02,154.21,142.80,141.71, 133.86,133.31,132.98,132.58,128.49(4C),127.47,127.10(3C),126.69, 124.49,120.91,58.88,57.60,54.16,51.81,49.13,49.07,47.48,43.92,42 .62,40.52,32.03,31.63,30.80(2C),22.62,17.16,11.57.HRMS(ESI)calcd for C 37 H 48 N 10 O2[M+H] + ,665.4034;found,665.4022.

[0164] Example 14: Compound YJZ1126

[0165] The synthesis method is similar to that of Example 1.

[0166] 1H NMR (600MHz, DMSO-d6) δ9.02(s,1H),8.72(s,1H),7.91(d,J=30.8Hz,2H),7.33–7.14(m,7H),6.94(d,J=8.3Hz,1H),6.66 (dd,J=16.9,10.3Hz,1H),6.28(dd,J=16.9,1.8Hz,1H),5.80(d,J=11.2Hz,1H),5.77–5.70(m,1H),4.81(p,J=6.7Hz,1H), 4.26(tt,J=12.0,3.6Hz,1H),4.18(d,J=6.0Hz,2H),3.64–3.47(m,1H),2.98–2.77(m,4H),2.47(t,J=10.5Hz,2H),2.36(s ,1H),2.25(s,3H),2.05–1.92(m,2H),1.79(t,J=9.4Hz,2H),1.46–1.33(m,8H),1.21–1.13(m,2H),1.01(d,J=6.2Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ163.63,160.36,157.04,154.18,153.88,142.83,1 41.70,133.86,133.10,132.98,132.58,128.50(4C),127.51,127.10(3C), 126.70,124.51,120.96,58.87,57.61,55.41,52.46,51.81,49.07,47.57, 43.92,42.62,40.50,31.67,30.82(2C),22.13(2C),17.17.HRMS(ESI)calcd for C 37 H 48 N 10 O2[M+H] + ,665.4034;found,665.4062.

[0167] Example 15: Compound YJZ1133

[0168] The synthesis method is similar to that of Example 1.

[0169] 1H NMR (600MHz, DMSO-d6) δ9.02(s,1H),8.71(s,1H),7.93(s,1H),7.83(s,1H),7.32(s,1H),7.30–7.25(m,2H),7.23(d,J=8.4Hz,1H) ,7.21–7.15(m,3H),6.94(d,J=7.9Hz,1H),6.66(dd,J=16.9,10.2Hz,1H),6.27(dd,J=17.0,1.8Hz,1H),5.84–5.70(m,2H),4.27(t, J=12.1Hz,1H),4.18(d,J=6.0Hz,2H),3.73–3.49(m,2H),2.98–2.76(m,4H),2.47(t,J=10.3Hz,2H),2.37(s,1H),2.26(s,3H),2.07 –1.85(m,2H),1.80(s,2H),1.42(q,J=11.8Hz,2H),1.22–1.13(m,2H),1.09–1.05(m,2H),1.01(d,J=6.2Hz,3H),0.99–0.94(m,2H). 13 C NMR(151MHz,DMSO-d6)δ163.64,160.55,157.04,154.24,142.83,141.70 ,133.85,133.12,132.95,132.57,128.50(4C),127.51,127.10(3C),126 .70,124.55,120.93,58.84,57.62,55.38,54.14,51.78,43.92,42.60,4 0.51,32.06,31.63,30.82(2C),28.57,17.14,6.07(2C).HRMS(ESI)calcd for C 37 H 46 N 10 O2[M+H] + ,663.3878;found,663.3914.

[0170] Example 16: Compound YJZ1127

[0171] The synthesis method is similar to that of Example 1.

[0172] 1H NMR (400MHz, DMSO-d6) δ9.02(s,1H),8.74(s,1H),7.91(d,J=15.6Hz,2H),7.36–7.10(m,7H),6.93(d,J=8.5Hz,1H),6.66(dd,J= 17.0,10.2Hz,1H),6.27(d,J=16.9Hz,1H),5.85–5.70(t,J=8.5Hz,2H),4.24(t,J=12.2Hz,1H),4.18(d,J=6.1Hz,2H),3.94(d,J =7.2Hz,2H),3.51(s,1H),2.96–2.75(m,4H),2.46(t,J=10.6Hz,2H),2.40–2.30(m,1H),2.25(s,3H),2.21–2.10(s,1H),2.03–1 .87(m,2H),1.79(d,J=9.6Hz,2H),1.40(q,J=12.3Hz,2H),1.17(q,J=12.3Hz,2H),1.00(d,J=6.2Hz,3H),0.80(d,J=6.7Hz,6H). 13 C NMR (151MHz, DMSO-d6) δ163.60,160.60,157.02,155.07,154.21,142.81,141. 72,133.90,133.31,132.97,132.60,128.49(3C),127.44,127.11(3C),126.69 ,124.48,120.88,58.91,57.59,54.17,53.15,51.81,49.82,49.36,43.93,42. 62,40.53,32.05,31.56,30.79(2C),28.89,20.33(2C),17.17.HRMS(ESI)calcd for C 38 H 50 N 10 O2[M+H] + ,679.4191;found,679.4234.

[0173] Example 17: Compound YJZ1128

[0174] The synthesis method is similar to that of Example 1.

[0175] 1H NMR (600MHz, DMSO-d6) δ9.03(s,1H),8.74(s,1H),7.91(d,J=29.6Hz,2H),7.30–7.16(m,7H),6.94(d,J=8.3Hz,1H),6.67(dd,J =16.9,10.2Hz,1H),6.28(dd,J=16.9,1.8Hz,1H),5.83–5.72(m,2H),4.26(tt,J=11.9,3.6Hz,1H),4.18(d,J=6.0Hz,2H),4.01( d,J=6.8Hz,2H),3.65–3.47(m,1H),2.98–2.74(m,4H),2.46(t,J=12.0Hz,2H),2.41–2.32(m,1H),2.03–1.86(m,2H),1.79(s,2H ),1.41(q,J=12.1Hz,2H),1.26–1.22(m,1H),1.21–1.11(m,2H),1.02(d,J=6.2Hz,3H),0.44(s,2H),0.34(q,J=3.2,1.7Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ163.63,160.61,157.03,154.23,142.83,141.70,133.90,133.23,132.96,132.58,128.50,127.49,127.10,126.70, 120.92,58.86,57.61,54.16,51.78,50.63,49.17,43.92,42.61,40.51,31.65,30.79(2C),29.47,17.15,11.35,4.01(2C).HRMS(ESI)calcd for C 38 H 48 N 10 O2[M+H] + ,677.4034;found,677.4074.

[0176] Example 18: Compound YJZ1145

[0177] The synthesis method is similar to that of Example 1.

[0178] 1H NMR (400MHz, DMSO-d6) δ9.04(s,1H),8.74(s,1H),7.91(d,J=23.3Hz,2H),7.35–7.19(m,4H),7.18–7.05(m,2H),6.96(d, J=8.9Hz,1H),6.66(dd,J=16.9,10.2Hz,1H),6.28(d,J=17.0Hz,1H),5.84–5.76(m,2H),4.32–4.17(m,3H),4.00(d,J=6. 9Hz,2H),3.52(s,2H),2.98–2.76(m,4H),2.48–2.43(m,2H),2.38(s,1H),2.26(s,3H),2.05–1.86(m,2H),1.79(d,J=10. 9Hz, 2H), 1.40 (q, J = 11.4Hz, 2H), 1.29–1.08 (m, 3H), 1.02 (d, J = 6.0Hz, 3H), 0.44 (d, J = 7.7Hz, 2H), 0.33 (q, J = 5.1Hz, 2H). 13 C NMR(151MHz,DMSO-d6)δ163.62,160.96(d,J=243.5Hz,1C),160.61,159.34,156.94(2C),154.57,154.21,142.89 ,133.75,133.22,133.00,132.59,128.96(d,J=4.8Hz,1C),128.56(d,J=8.1Hz,1C),128.36(d,J=14.4Hz,1C),12 7.48(2C),124.62(d,J=3.1Hz,1C),124.46,120.93,115.22(d,J=21.1Hz,1C),58.89,57.60,54.24,51.81,50.60 ,49.15,42.63,40.53,37.67(d,J=4.9Hz,1C),32.04,31.62,30.76(2C),17.18,11.35,4.01(2C).HRMS(ESI)calcd for C 38 H 47 N 10 O2F[M+H] + ,695.3940;found,695.3926.

[0179] Example 19: Compound YJZ1146

[0180] The synthesis method is similar to that of Example 1.

[0181] 1 H NMR (600MHz, DMSO-d6) δ9.27(s,1H),8.74(s,1H),7.97(s,1H),7.89(d,J=10.3Hz,2H),7.36–7.26(m,3H),7.19(dd,J=7.9,2.4Hz,3H),6.70(dd,J=16 .9,10.3Hz,1H),6.30(dd,J=17.0,1.9Hz,1H),6.23(s,1H),5.83–5.78(m, 1H),4.29–4.23(m,1H),4.18(d,J=6.0Hz,2H),4.01(d,J=7.1Hz,2H),3.53( s,1H),3.40(d,J=11.6Hz,2H),2.97(td,J=11.9,2.7Hz,1H),2.77(dt,J=1 1.4,2.8Hz,1H),2.59(d,J=22.2Hz,1H),2.43(t,J=10.3Hz,1H),2.39–2.30 (m,1H),2.05–1.88(m,3H),1.85–1.73(m,2H),1.46–1.36(m,2H),1.21–1. 08(m,3H),1.01(d,J=6.2Hz,3H),0.44(d,J=7.8Hz,2H),0.36–0.31(m,2H).

[0182] Example 20: Compound YJZ1148

[0183] The synthesis method is similar to that of Example 1.

[0184] 1H NMR (600MHz, DMSO-d6) δ9.29 (s, 1H), 8.74 (s, 1H), 7.93 (t, J = 27.7Hz, 3H), 7.36 –7.19(m,4H),7.16(td,J=7.4,1.2Hz,1H),7.14–7.06(m,1H),6.70(dd,J=17.0, 10.2Hz,1H),6.30(dd,J=17.0,1.9Hz,1H),6.22(s,1H),5.80(dd,J=10.1,1.9H z,1H),4.30–4.19(m,4H),4.02(dd,J=9.3,5.5Hz,2H),3.53(s,1H),3.40(t,J=1 1.4Hz,2H),2.97(td,J=11.9,2.7Hz,1H),2.77(dt,J=11.2,2.9Hz,1H),2.59(d d,J=12.3,10.0Hz,1H),2.42(td,J=11.7,3.1Hz,1H),2.38–2.30(m,1H),2.24(s ,3H),2.04–1.87(m,3H),1.87–1.71(m,2H),1.48–1.36(m,2H),1.18(t,J=7.1Hz ,3H),1.01(d,J=6.2Hz,4H),0.45(t,J=6.3Hz,2H),0.34(dt,J=6.3,4.3Hz,2H).

[0185] Example 21: Compound ZLC-8-103

[0186] The synthesis method is similar to that of Example 1.

[0187] 1H NMR (600MHz, DMSO-d6) δ9.26 (s, 1H), 9.05 (s, 1H), 7.96 (s, 1H), 7.82 (d, J = 2. 4Hz,1H),7.74(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.9Hz,1H),7.41(d,J=8.5Hz ,1H),7.25(d,J=8.1Hz,1H),7.17(ddd,J=8.0,6.9,1.1Hz,1H),6.74(dd,J=1 7.0,10.2Hz,1H),6.29(dd,J=17.0,1.9Hz,1H),5.80(dd,J=10.2,1.9Hz,1H), 5.52–5.46(m,1H),4.24(tt,J=12.0,3.6Hz,1H),3.60(d,J=7.8Hz,1H),3.55 (d,J=12.7Hz,2H),2.84–2.75(m,4H),2.37(s,6H),1.98(d,J=11.4Hz,2H),1. 90(d,J=11.8Hz,2H),1.78–1.74(m,4H),1.65–1.59(m,2H),1.59–1.51(m,3H) ,1.46–1.38(m,2H),1.37–1.30(m,1H),1.16–1.05(m,5H),0.78–0.71(m,2H).

[0188] Example 22: Compound ZLC-10-36

[0189] The synthesis method is similar to that of Example 1.

[0190] 1H NMR(600MHz,DMSO-d6)δ9.50(s,1H),9.05(s,1H),7.73(d,J=7.9Hz,1H),7.62(t,J=7.8Hz,1H),7.42(d,J=8.7Hz,1H),7 .28–7.23(m,4H),7.20–7.15(m,4H),6.93(d,J=3.0Hz,2H),6.65(dd,J=17.1,10.2Hz,1H),6.26(dd,J=16.9,2.0Hz,1H), 5.77–5.71(m,1H),5.60–5.58(m,1H),4.31–4.26(m,1H),4.19(d,J=6.2Hz,2H),3.65–3.56(m,2H),3.29–3.25(m,3H),2 .88(s,1H),2.27(s,6H),2.16–2.09(m,1H),2.01–1.95(m,2H),1.84–1.76(m,3H),1.46–1.39(m,2H),1.21–1.13(m,2H).

[0191] Example 23: Compound ZLC-10-37

[0192] The synthesis method is similar to that of Example 1.

[0193] 1H NMR (600MHz, DMSO-d6) δ10.21(s,1H),9.04(d,J=8.2Hz,1H),8.27(d,J=2.4Hz,1H),7.73(dd,J=8.0,1.5Hz,1H),7.61(t,J=7.7Hz,1H),7.40( d,J=8.6Hz,1H),7.34(d,J=8.4Hz,1H),7.29–7.23(m,3H),7.20–7.14(m,4H),6.91(dd,J=8.4,2.5Hz,1H),6.45(dd,J=16.9,10.1Hz,1H),6.3 1(dd,J=16.9,2.0Hz,1H),5.83–5.80(m,1H),5.75(t,J=6.2Hz,1H),4.31–4.26(m,1H),4.19(d,J=6.0Hz,2H),3.64–3.58(m,1H),2.84(t,J=5 .5Hz,2H),2.72(s,3H),2.42(t,J=5.5Hz,2H),2.24(s,6H),1.97(s,2H),1.85–1.76(m,2H),1.43(qd,J=13.0,3.4Hz,2H),1.25–1.18(m,2H).

[0194] Example 24: Compound ZLC-10-40

[0195] The synthesis method is similar to that of Example 1.

[0196] 1H NMR (600MHz, DMSO-d6) δ9.13(s,1H),9.05(s,1H),7.98(s,1H),7.74(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.7Hz,1H),7.41 (d,J=8.6Hz,1H),7.29–7.23(m,4H),7.20–7.15(m,4H),6.96(dd,J=8.4,2.5Hz,1H),6.74(dd,J=16.9,10.2Hz,1H),6.2 9(dd,J=17.0,1.9Hz,1H),5.83–5.77(m,2H),4.28(tt,J=12.0,3.7Hz,1H),4.18(d,J=6.0Hz,2H),3.83(t,J=4.5Hz,4H) ,3.64–3.55(m,1H),2.90–2.84(m,4H),1.97(d,J=12.4Hz,2H),1.83–1.77(m,2H),1.46–1.39(m,2H),1.23–1.15(m,2H).

[0197] Example 25: Compound ZLC-10-41

[0198] The synthesis method is similar to that of Example 1.

[0199] 1H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.03(s,1H),7.95(s,1H),7.73(dd,J=8.1,1.5Hz,1H),7.62(t,J=7.6Hz,1H),7.41(d,J=8.3Hz,1H), 7.29–7.22(m,4H),7.20–7.15(m,4H),6.95(dd,J=8.4,2.5Hz,1H),6.68(dd,J=17.0,10.3Hz,1H),6.28(dd,J=16.9,1.8Hz,1H),5.81–5.76 (m,2H),4.57(q,J=5.7,4.9Hz,2H),4.49(t,J=6.1Hz,2H),4.28(tt,J=12.0,3.6Hz,1H),4.18(d,J=6.0Hz,2H),3.64–3.56(m,1H),3.53(p ,J=6.3Hz,1H),2.91(d,J=4.9Hz,4H),2.55–2.49(m,4H),2.01–1.92(m,2H),1.80(d,J=11.5Hz,2H),1.46–1.38(m,2H),1.22–1.15(m,2H).

[0200] Example 26: Compound ZLC-10-44

[0201] The synthesis method is similar to that of Example 1.

[0202] 1H NMR (600MHz, DMSO-d6) δ9.49(s,1H),9.05(s,1H),7.77–7.71(m,1H),7.63(t,J=7.8Hz,1H),7.43(d,J=8.7Hz,1H),7.31–7 .21(m,4H),7.19–7.17(m,4H),6.92(s,2H),6.61(dd,J=17.0,10.3Hz,1H),6.28–6.23(m,1H),5.78–5.71(m,1H),5.60–5. 58(m,1H),4.31–4.26(m,1H),4.23–4.16(m,2H),3.63–3.59(m,1H),3.37–3.32(m,1H),3.25(dd,J=16.0,8.0Hz,3H),2.78 (s,1H),2.22(s,6H),2.14–2.09(m,1H),1.97(d,J=11.9Hz,2H),1.84–1.72(m,3H),1.47–1.39(m,2H),1.23–1.12(m,2H).

[0203] Example 27: Compound ZLC-11-22

[0204] The synthesis method is similar to that of Example 1.

[0205] 1H NMR (600MHz, DMSO-d6) δ9.27(d,J=3.1Hz,1H),9.04(s,1H),7.99(d,J=2.4Hz,1H),7.93(d,J=2.4Hz,1H),7.74(dd,J=8.0,1.5Hz,1H),7.62(t,J =7.5Hz,1H),7.39(d,J=8.5Hz,1H),7.30–7.22(m,5H),7.19–7.16(m,2H),5.92(d,J=8.0Hz,1H),5.79(dd,J=48.6,3.8Hz,1H),5.53(dd,J=15.8 ,3.8Hz,1H),4.86(p,J=7.2Hz,1H),4.27–4.22(m,1H),3.62–3.56(m,1H ),3.56–3.51(m,2H),2.82(t,J=12.2Hz,2H),2.44–2.35(m,1H),2.30(s ,6H),2.01–1.90(m,4H),1.84–1.80(m,1H),1.75–1.72(m,1H),1.59–1. 53(m,2H),1.47–1.37(m,2H),1.28(d,J=7.1Hz,3H),1.16–1.05(m,2H).

[0206] Example 28: Compound ZLC-11-30

[0207] The synthesis method is similar to that of Example 1.

[0208] 1H NMR (600MHz, DMSO-d6) δ11.94(d,J=2.7Hz,1H),9.04(s,1H),8.09(d,J=2.1Hz,1H),7.75–7.71(m,1H),7.63–7.60(m,1H),7.38(d,J=8.5Hz,1H ),7.33(d,J=8.0Hz,1H),7.29–7.21(m,3H),7.17(ddd,J=8.0,6.9,1.1Hz,1H),7.10–7.06(m,2H),6.92(dd,J=7.9,2.1Hz,1H),5.77(dd,J=48. 7,3.7Hz,1H),5.50(dd,J=15.7,3.7Hz,1H),5.42(d,J=7.9Hz,1H),4.84(p,J=7.2Hz,1H),4.27–4.22(m,1H),3.73(s,2H),3.64(s,4H),3.61–3 .54(m,1H),2.50–2.43(m,4H),1.95(s,2H),1.83–1.80(m,1H),1.76–1. 70(m,1H),1.45–1.36(m,2H),1.25(d,J=7.1Hz,3H),1.21–1.12(m,2H).

[0209] Example 29: Compound ZLC-11-31

[0210] The synthesis method is similar to that of Example 1.

[0211] 1H NMR (600MHz, DMSO-d6) δ11.97(s,1H),9.04(s,1H),8.09(d,J=2.1Hz,1H),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.9Hz,1H),7.39(d,J=8.5Hz,1H ),7.32(d,J=8.0Hz,1H),7.29–7.22(m,3H),7.17(td,J=7.4,6.8,1.1Hz, 1H),7.11–7.05(m,2H),6.91(dd,J=7.9,2.1Hz,1H),5.76(dd,J=48.4,3. 6Hz, 1H), 5.50 (dd, J=15.5, 3.6Hz, 1H), 5.43 (d, J=7.9Hz, 1H), 4.84 (p, J= 7.2Hz,1H),4.24(tt,J=12.1,3.6Hz,1H),3.71(s,2H),3.62–3.55(m,1H) ,3.34(s,6H),2.19(s,3H),1.96(d,J=11.3Hz,2H),1.83–1.80(m,1H),1. 75–1.72(m,1H),1.46–1.36(m,2H),1.26–1.22(m,5H),1.21–1.13(m,2H).

[0212] Example 30: Compound ZLC-11-32

[0213] The synthesis method is similar to that of Example 1.

[0214] 1H NMR(600MHz,DMSO-d6)δ9.37(d,J=4.0Hz,1H),9.04(s,1H),8.04(d,J=2.4Hz,1H) ,7.73(dd,J=8.0,1.5Hz,1H),7.61(t,J=7.8Hz,1H),7.38(d,J=8.5Hz,1H),7.31( d,J=8.4Hz,1H),7.28–7.23(m,3H),7.17(ddd,J=8.0,6.8,1.1Hz,1H),7.11–7.05 (m,2H),6.96(dd,J=8.3,2.5Hz,1H),5.79(dd,J=49.3,3.8Hz,1H),5.53(dd,J=16. 0,3.8Hz,1H),5.44(d,J=8.0Hz,1H),4.84(p,J=7.2Hz,1H),4.26–4.21(m,1H),3. 60–3.53(m,1H),3.10–3.04(m,2H),2.74(t,J=11.6Hz,2H),2.60–2.53(m,2H),2. 45–2.30(m,4H),2.21(s,3H),2.00–1.90(m,4H),1.84–1.77(m,1H),1.73–1.70(m ,1H),1.60–1.52(m,2H),1.45–1.36(m,2H),1.27–1.21(m,5H),1.17–1.09(m,2H).

[0215] Example 31: Compound ZLC-11-34

[0216] The synthesis method is similar to that of Example 1.

[0217] 1H NMR (600MHz, DMSO-d6) δ11.99(s,1H),9.03(s,1H),8.09(d,J=2.1Hz,1H),7.73(dd,J=8.0,1.5Hz,1H),7.61(t,J=7.8Hz,1H),7.38(d,J=8.5Hz,1H ),7.32(d,J=8.0Hz,1H),7.29–7.21(m,3H),7.16(ddd,J=8.0,6.8,1.1Hz ,1H),7.10–7.05(m,2H),6.91(dd,J=7.9,2.1Hz,1H),5.75(dd,J=48.3,3. 6Hz, 1H), 5.49 (dd, J=15.6, 3.6Hz, 1H), 5.42 (d, J=8.0Hz, 1H), 4.84 (p, J= 7.2Hz,1H),4.27–4.22(m,1H),3.71(s,2H),3.63–3.54(m,1H),2.76–2.5 2(m,2H),2.48–2.10(m,6H),1.99–1.94(m,2H),1.86–1.80(m,1H),1.75– 1.72(m,1H),1.45–1.36(m,2H),1.28–1.13(m,7H),0.99(t,J=7.1Hz,3H).

[0218] Example 32: Compound ZLC-11-70

[0219] The synthesis method is similar to that of Example 1.

[0220] 1H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.02(s,1H),7.95(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.41(d,J=8.5Hz, 1H),7.29–7.22(m,4H),7.18–7.16(m,4H),6.93(dd,J=8.3,2.5Hz,1H),6.68(dd,J=16.9,10.3Hz,1H),6.28(dd,J=17.0,1.8Hz,1H), 5.82–5.76(m,2H),4.30–4.17(m,1H),4.17(d,J=6.0Hz,2H),3.63–3.56(m,1H),2.90–2..84(m,4H),2.81–2.79(m,2H),2.71–2.69(m ,4H),2.21–2.15(m,1H),2.13(s,3H),2.01–1.92(m,2H),1.87–1.82(m,2H),1.81–1.73(m,4H),1.48–1.39(m,4H),1.22–1.15(m,2H).

[0221] Example 33: Compound ZLC-11-71

[0222] The synthesis method is similar to that of Example 1.

[0223] 1H NMR (600MHz, DMSO-d6) δ9.04(s,1H),8.99(s,1H),7.96(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.41(d,J=8.5Hz,1H),7.31 –7.25(m,2H),7.23(d,J=8.0Hz,1H),7.21–7.15(m,5H),6.92(dd,J=8.3,2.5Hz,1H),6.75(dd,J=16.9,10.3Hz,1H),6.28(dd,J=17.0,1.9Hz,1H ),5.80–5.75(m,2H),4.30–4.25(m,1H),4.17(d,J=6.0Hz,2H),3.64–3 .54(m,1H),3.41(s,2H),3.07(d,J=11.3Hz,2H),2.68–2.64(m,2H),2.6 0–2.51(m,3H),2.43–2.23(m,4H),2.15(s,3H),2.02–1.93(m,2H),1.90 –1.83(m,2H),1.82–1.70(m,4H),1.46–1.38(m,2H),1.22–1.14(m,2H).

[0224] Example 34: Compound ZLC-11-72

[0225] The synthesis method is similar to that of Example 1.

[0226] 1H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.01(s,1H),7.96(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.61(t,J=7.7Hz,1H),7.41(d,J=8.5Hz,1H),7.26(dd,J= 8.4,6.8Hz,3H),7.23(d,J=8.1Hz,1H),7.18–7.16(m,4H),6.92(dd,J=8.3 ,2.5Hz,1H),6.71(dd,J=16.9,10.2Hz,1H),6.28(dd,J=17.0,1.8Hz,1H),5 .79(dd,J=10.2,1.8Hz,1H),5.74(d,J=6.5Hz,1H),4.30–4.26(m,1H),4.1 7(d,J=6.0Hz,2H),3.64–3.55(m,1H),2.81(t,J=5.5Hz,4H),2.30–2.26(m, 4H),2.15(s,3H),2.01–1.95(m,2H),1.80(dd,J=10.7,4.7Hz,2H),1.64(t, J=5.4Hz,4H),1.52(t,J=5.6Hz,4H),1.45–1.38(m,2H),1.21–1.14(m,2H).

[0227] Example 35: Compound ZLC-11-73

[0228] The synthesis method is similar to that of Example 1.

[0229] 1H NMR(600MHz,DMSO-d6)δ9.23(s,1H),9.04(s,1H),8.00–7.91(m,1H),7.85(d,J=2.4Hz,1H),7.74(dd,J=8.0,1.5Hz,1H),7.63–7.61(m,1H),7.45– 7.38(m,1H),7.23(d,J=8.1Hz,1H),7.20–7.14(m,3H),7.09–7.04(m,2H) ,6.76(dd,J=17.0,10.3Hz,1H),6.56(s,1H),6.31(dd,J=16.9,2.0Hz,1H ),5.81(dd,J=10.2,1.9Hz,1H),4.23–4.18(m,1H),3.58(d,J=16.4Hz,1H ),3.52(d,J=12.5Hz,2H),2.76(td,J=12.4,2.3Hz,2H),2.22–2.18(m,7H ),2.00–1.91(m,2H),1.84(d,J=12.0Hz,2H),1.77(d,J=12.4Hz,2H),1.7 2–1.67(m,2H),1.46–1.37(m,2H),1.15–1.08(m,2H),1.07–1.04(m,4H).

[0230] Example 36: Compound ZLC-11-74

[0231] The synthesis method is similar to that of Example 1.

[0232] 1H NMR (600MHz, DMSO-d6) δ9.26(d,J=3.2Hz,1H),9.04(s,1H),7.96(d,J=2.4Hz,1H),7.92(d,J=2.4Hz,1H),7.73(dd,J=7.9,1.5Hz,1H),7.62( t,J=7.5Hz,1H),7.39(d,J=8.5Hz,1H),7.30–7.22(m,3H),7.18–7.16(m,1H),7.11–7.06(m,2H),5.97(d,J=8.0Hz,1H),5.85–5.73(m,1H),5 .53(dd,J=15.8,3.9Hz,1H),4.86(p,J=7.2Hz,1H),4.26–4.20(m,1H),3.63–3.50(m,6H),2.82(td,J=12.5,2.2Hz,2H),2.66(s,4H),2.49–2 .42(m,1H),2.37(s,3H),2.02–1.88(m,4H),1.85–1.69(m,2H),1.60– 1.54(m,2H),1.47–1.36(m,2H),1.29–1.21(m,4H),1.16–1.03(m,2H).

[0233] Example 37: Compound ZLC-11-75

[0234] The synthesis method is similar to that of Example 1.

[0235] 1H NMR (600MHz, DMSO-d6) δ12.06 (s, 1H), 9.04 (s, 1H), 8.09 (d, J = 2.1Hz, 1H), 7.73 (dd,J=8.0,1.5Hz,1H),7.62(t,J=7.9Hz,1H),7.39(d,J=8.5Hz,1H),7.31(d,J= 8.0Hz,1H),7.29–7.25(m,2H),7.24(d,J=7.9Hz,1H),7.19–7.16(m,1H),7.11– 7.06(m,2H),6.92(dd,J=8.0,2.2Hz,1H),5.76(dd,J=48.6,3.6Hz,1H),5.49(dd ,J=15.7,3.7Hz,1H),5.46–5.41(m,1H),4.84(p,J=7.2Hz,1H),4.27–4.21(m,1 H),3.71(s,2H),3.63–3.54(m,1H),2.94(d,J=11.3Hz,2H),2.36(s,6H),2.07(t ,J=11.6Hz,2H),2.01–1.92(m,2H),1.88(d,J=11.6Hz,2H),1.84–1.70(m,2H), 1.53–1.46(m,2H),1.45–1.36(m,2H),1.25(d,J=7.1Hz,3H),1.23–1.12(m,2H).

[0236] Example 38: Compound ZLC-11-76

[0237] The synthesis method is similar to that of Example 1.

[0238] 1H NMR(600MHz,DMSO-d6)δ9.42(d,J=4.1Hz,1H),9.04(s,1H),8.04(d,J=2.4Hz,1H ),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.38(d,J=8.5Hz,1H),7.35 (d,J=8.4Hz,1H),7.29–7.21(m,3H),7.18–7.15(m,1H),7.11–7.05(m,2H),6.98( dd,J=8.3,2.4Hz,1H),5.79(dd,J=49.2,3.8Hz,1H),5.52(dd,J=15.9,3.8Hz,1H) ,5.49(d,J=7.8Hz,1H),4.84(p,J=7.2Hz,1H),4.26–4.21(m,1H),3.62–3.51(m, 1H),2.97(d,J=11.1Hz,2H),2.91(d,J=4.9Hz,4H),2.67(s,4H),2.29(d,J=16.9H z,4H),2.18(s,2H),2.02–1.89(m,2H),1.84–1.79(m,3H),1.74–1.70(m,1H),1.5 4(q,J=12.1Hz,2H),1.45–1.35(m,2H),1.24(d,J=7.1Hz,3H),1.19–1.10(m,2H).

[0239] Example 39: Compound ZLC-12-6

[0240] The synthesis method is similar to that of Example 1.

[0241] 1H NMR(600MHz, DMSO-d6)δ9.04(s,1H),9.02(s,1H),7.95(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.65–7.60(m,1H),7.41( d,J=8.6Hz,1H),7.29–7.23(m,4H),7.20–7.14(m,4H),6.94(dd,J=8.4,2.5Hz,1H),6.68(dd,J=17.0,10.3Hz,1H),6. 28(dd,J=17.0,1.8Hz,1H),5.80–5.77(m,2H),4.31–4.26(m,1H),4.18(d,J=6.0Hz,2H),3.63–3.59(m,1H),2.89–2.8 5(m,4H),2.62–2.53(m,4H),2.27(s,3H),1.98–1.95(m,2H),1.85–1.76(m,2H),1.45–1.39(m,2H),1.23–1.15(m,2H).

[0242] Example 40: Compound ZLC-12-7

[0243] The synthesis method is similar to that of Example 1.

[0244] 1 H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.02(s,1H),7.96(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.65–7.60(m,1H),7.41(d,J=8.6Hz ,1H),7.29–7.23(m,4H),7.20–7.15(m,4H),6.94(dd,J=8.3,2.5Hz,1H),6.68(dd,J=16.9,10.2Hz,1H),6.28(dd,J=17.0,1.8Hz ,1H),5.82–5.76(m,2H),4.31–4.26(m,1H),4.18(d,J=6.0Hz,2H),3.63–3.57(m,1H),2.89–2.85(m,4H),2.72–2.53(m,4H),2.4 2(q,J=7.2Hz,2H),2.03–1.90(m,2H),1.80(dd,J=10.4,4.6Hz,2H),1.47–1.38(m,2H),1.24–1.15(m,2H),1.04(t,J=7.2Hz,3H).

[0245] Example 41: Compound ZLC-12-8

[0246] The synthesis method is similar to that of Example 1.

[0247] 1 H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.00(s,1H),7.97(d,J=5.4Hz,1H),7.73(dd,J=8.0,1.5Hz,1H),7.65–7.58(m,1H),7.41(d,J=8.6H z,1H),7.29–7.22(m,3H),7.21–7.15(m,5H),6.92(dd,J=8.4,2.5Hz,1H),6.74(dd,J=16.9,10.3Hz,1H),6.29(dd,J=16.9,1.8Hz,1H),5. 81–5.74(m,2H),4.31–4.25(m,1H),4.18(d,J=6.0Hz,2H),3.64–3.56(m,5H),3.08(d,J=11.2Hz,2H),2.68–2.62(m,2H),2.54–2.51(m,3 H),2.28–2.23(m,1H),2.03–1.95(m,2H),1.92–1.85(m,2H),1.83–1.77(m,2H),1.74–1.70(m,2H),1.47–1.38(m,2H),1.27–1.15(m,3H).

[0248] Example 42: Compound ZLC-12-9

[0249] The synthesis method is similar to that of Example 1.

[0250] 1H NMR (600MHz, DMSO-d6) δ9.04(s,1H),9.03(s,1H),7.95(s,1H),7.73(dd,J=8.0,1.5Hz,1H),7.63–7.60(m,1H),7.41(d,J=8.5Hz, 1H),7.29–7.23(m,4H),7.20–7.15(m,4H),6.94(dd,J=8.4,2.5Hz,1H),6.68(dd,J=16.9,10.3Hz,1H),6.28(dd,J=17.0,1.8Hz,1H ),5.80–5.77(m,2H),4.31–4.25(m,1H),4.18(d,J=6.0Hz,2H),3.94–3.88(m,2H),3.62–3.56(m,1H),3.32–3.27(m,2H),2.89–2. 86(m,4H),2.77–2.67(m,4H),2.49–2.42(m,1H),1.97(d,J=11.7Hz,2H),1.83–1.73(m,4H),1.47–1.38(m,4H),1.23–1.14(m,2H).

[0251] Example 43: Compound ZLC-12-20

[0252] The synthesis method is similar to that of Example 1.

[0253] 1H NMR (600MHz, DMSO-d6) δ9.39(d,J=4.0Hz,1H),9.04(s,1H),8.04(d,J=2.4Hz,1H),7.75–7.71(m,1H),7.62(t,J=7.7Hz,1H),7.38(dd,J=8.6, 6.2Hz,2H),7.28–7.23(m,3H),7.19–7.15(m,1H),7.11–7.05(m,2H),6.99(dd,J=8.4,2.4Hz,1H),5.78(dd,J=49.2,3.8Hz,1H),5.51(dd,J=1 5.9,3.8Hz,1H),5.49–5.45(m,1H),4.85(p,J=7.2Hz,1H),4.60–4.57( m,2H),4.49–4.46(m,2H),4.27–4.22(m,1H),3.61–3.52(m,1H),3.51– 3.47(m,1H),2.96–2.92(m,4H),2.45–2.38(m,3H),1.95(s,2H),1.83– 1.71(m,2H),1.46–1.36(m,2H),1.27–1.22(m,4H),1.21–1.09(m,2H).

[0254] Example 44: Compound ZLC-12-22

[0255] The synthesis method is similar to that of Example 1.

[0256] 1H NMR(600MHz,DMSO-d6)δ9.27(s,1H),9.04(s,1H),7.96(s,1H),7.87(d,J=2.4Hz ,1H),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.45–7.38(m,1H),7.3 1–7.28(m,1H),7.25–7.21(m,2H),7.17(ddd,J=8.0,6.8,1.1Hz,1H),7.13(td,J =7.5,1.2Hz,1H),7.09(ddd,J=10.8,8.2,1.2Hz,1H),6.75(dd,J=17.0,10.3Hz,1 H),6.31(dd,J=17.0,1.9Hz,1H),5.96(d,J=8.0Hz,1H),5.81(dd,J=10.2,1.9Hz ,1H),5.11(p,J=7.2Hz,1H),4.23–4.18(m,1H),3.62–3.55(m,3H),2.80–2.76(m, 2H),2.54–2.51(m,1H),2.26–2.20(m,7H),2.02–1.96(m,2H),1.87–1.78(m,3H) ,1.74–1.65(m,3H),1.45–1.35(m,2H),1.25(d,J=7.1Hz,3H),1.18–1.03(m,2H).

[0257] Example 45: Compound ZLC-12-45

[0258] The synthesis method is similar to that of Example 1.

[0259] 1H NMR (600MHz, DMSO-d6) δ9.27(s,1H),9.04(s,1H),7.98–7.92(m,2H),7.73(dd,J=8.0,1.5Hz,1H),7.62(t,J=7.5Hz,1H),7.39(d,J=8.5Hz,1H),7.3 0–7.26(m,2H),7.25(d,J=7.9Hz,1H),7.19–7.15(m,1H),7.11–7.06(m,2H ),5.99(d,J=8.0Hz,1H),5.78(dd,J=48.7,3.8Hz,1H),5.52(dd,J=15.8,3 .9Hz,1H),4.86(p,J=7.2Hz,1H),4.23(tt,J=12.1,3.7Hz,1H),3.63–3.53 (m,1H),3.22–3.10(m,4H),2.83–2.80(m,2H),2.66–2.61(m,4H),2.21–2. 16(m,4H),2.01–1.95(m,2H),1.91–1.87(m,2H),1.83–1.80(m,1H),1.79– 1.70(m,3H),1.49–1.37(m,4H),1.27(d,J=7.1Hz,3H),1.16–1.04(m,2H).

[0260] Example 46: Compound ZLC-12-47

[0261] The synthesis method is similar to that of Example 1.

[0262] 1H NMR (600MHz, DMSO-d6) δ11.95(s,1H),9.04(s,1H),8.08(t,J=1.8Hz,1H),7.73(d d,J=8.1,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.39(d,J=8.5Hz,1H),7.33(d,J=8.0H z,1H),7.29–7.25(m,2H),7.25–7.21(m,1H),7.17(ddd,J=8.0,6.8,1.1Hz,1H),7 .10–7.05(m,2H),6.92(dd,J=7.9,2.1Hz,1H),5.75(dd,J=48.5,3.7Hz,1H),5.49( dd,J=15.6,3.6Hz,1H),5.44(d,J=8.0Hz,1H),4.84(p,J=7.2Hz,1H),4.54–4.52( m,2H),4.43–4.41(m,2H),4.26–4.21(m,1H),3.73(s,2H),3.63–3.53(m,1H),3.39 (p,J=6.3Hz,1H),2.71–2.51(m,2H),2.47–2.29(m,2H),1.95(s,2H),1.83–1.79(m ,1H),1.75–1.71(m,1H),1.45–1.36(m,2H),1.29–1.22(m,7H),1.20–1.12(m,2H).

[0263] Example 47: Compound ZLC-12-48

[0264] The synthesis method is similar to that of Example 1.

[0265] 1H NMR (600MHz, DMSO-d6) δ9.30 (s, 1H), 9.04 (s, 1H), 7.95 (d, J = 2.3Hz, 1H), 7.93 (d ,J=2.5Hz,1H),7.73(dd,J=8.1,1.5Hz,1H),7.62(t,J=7.8Hz,1H),7.39(d,J=8. 5Hz,1H),7.30–7.27(m,2H),7.25(d,J=7.6Hz,1H),7.17(ddd,J=8.0,6.9,1.1Hz ,1H),7.11–7.07(m,2H),5.99(d,J=8.0Hz,1H),5.78(dd,J=48.6,3.9Hz,1H),5. 51(dd,J=15.8,3.8Hz,1H),4.86(p,J=7.3Hz,1H),4.58–4.56(m,2H),4.49–4.46 (m,2H),4.26–4.21(m,1H),3.58(d,J=10.5Hz,1H),3.49(p,J=6.3Hz,1H),3.24– 3.20(m,4H),2.47–2.44(m,4H),1.96–1.91(m,2H),1.85–1.79(m,1H),1.74(dd, J=13.0,4.1Hz,1H),1.46–1.36(m,2H),1.27(d,J=7.1Hz,3H),1.17–1.03(m,2H).

[0266] Example 48: Activity inhibition test of compounds on CDK12 and CDK13 kinases

[0267] The kinase inhibitory activity of the test compounds was determined by ADP-Glo TM Kinase Assay The assay was evaluated by detecting the conversion of ADP formed in the kinase reaction to ATP by Ultra-Glo TM Luminescence is converted by luciferase into a luminescent signal, which reflects kinase activity. Luminescence is positively correlated with kinase activity. A simplified protocol is outlined below.

[0268] 1. Prepare 2× ATP / substrate solution and 2× kinase solution using kinase reaction buffer (HEPES, Brij35, EGTA, MgCl2, DTT). CDK12 / Cyclin K, CDK13 / Cyclin K kinase ( H24A) working concentrations were 50nM and 80nM, respectively, and ATP ( V915B) working concentrations were 8 μM and 4 μM respectively. The kinase reaction substrate used was PS7-CTD ( PE0401), with a working concentration of 0.1 mg / mL.

[0269] 2. The compound test concentration was 10 μM starting point, 3-fold dilution of 10 concentration points. 655 Transfer 40 nL of compound dilution to the 384 assay plate, centrifuge, add 2 μL of 2× kinase solution to the 384 assay plate, and centrifuge at 1000 rpm for 1 minute.

[0270] 3. After incubation at 25°C for 10 minutes, add 2 μL of 2× substrate and ATP solution to the 384 assay plate and centrifuge at 1000 rpm for 1 minute.

[0271] 4. After incubation at 25°C for 60 minutes, add 4 μL of ADP-Glo ​​reagent to the 384 assay plate and centrifuge at 1000 rpm for 1 minute.

[0272] 5. After incubation at 25°C for 40 minutes, add 8 μL of kinase assay reagent to the 384-well plate and centrifuge at 1000 rpm for 1 minute.

[0273] 6. Incubate at 25°C for 40 minutes and then read the luminescence signal using a BMG Microplate Reader.

[0274] The percentage of kinase inhibition of the compound was calculated as (negative control value - compound well value) ÷ (negative control value - positive control value) × 100. The IC was obtained by fitting the sigmoidal dose-response curve of the compound. 50 All fittings were performed using GraphPad Prism software (GraphPad Software Inc.). The results of the kinase activity test are shown in Table 1.

[0275] Table 1 Test results of kinase inhibitory activity of compounds (IC 50 :nM)

[0276] IC 50 :<20nM=*; 20-50nM=**; 50-500nM=***; >500nM=****.

[0277] It can be seen from the data in Table 1 that the novel CDK12 / 13 covalent inhibitor substituted with a cyclic structure of the present invention has a strong inhibitory activity against CDK12 / 13 kinases.

[0278] Example 49 Study on the Inhibitory Activity of Breast Cancer Cell Proliferation

[0279] The cell proliferation inhibitory activity of the compound was determined using Cell Counting Kit-8 Cell Viability Assay (Selleck.cn). Breast cancer HCC38 cells were inoculated in RPMI 1640 medium. 96-well plates Incubate in a 37°C, 5% CO2 incubator. After overnight incubation, prepare a high concentration stock solution of each test compound with culture medium, and add the compound to a 96-well plate according to a certain concentration gradient. After the administration is completed, the cell plate is placed in a 37°C, 5% CO2 incubator for 3 days. Then, add 15 μL of cck-8 reagent to each well of the 96-well plate, and return the plate to the incubator for incubation for 1.5-2 hours. After the incubation is completed, remove the plate, centrifuge to remove bubbles, and then use The absorbance values ​​of each well at 450 nm and 650 nm were obtained using GraphPad Prism software (PerkinElmer) and the data were processed and analyzed using GraphPad Software Inc. The test results are shown in Table 2.

[0280] Table 2 Breast cancer cell activity test results of compounds (IC 50 :nM)

[0281] IC 50 :<20nM=*; 20-50nM=**; 50-500nM=***; >500nM=****.

[0282] It can be seen from the data in Table 2 that the novel CDK12 / 13 covalent inhibitor compound containing a cyclic structure of the present invention has a strong inhibitory activity on the proliferation of HCC38 cells.

[0283] Example 50 Study on Prostate Cancer Cell Proliferation Inhibition Activity

[0284] The cell counting kit-8 cell viability assay (Selleck.cn) was used to determine the inhibitory activity of the compound on prostate cancer cell proliferation. 384-well plates After overnight incubation, prepare a high concentration stock solution of each test compound; use 650Liquid Handler The compound was added to a 384-well plate according to a certain concentration gradient. After the administration, the cell plate was placed in an incubator at 37°C with 5% CO2 for 3 days. (Thermo Scientific TM ) Add 5 μL of cck-8 reagent to each well of the 384-well plate and return the plate to the incubator for 1.5-2 hours. After incubation, remove the plate and centrifuge to remove bubbles before use. The absorbance values ​​of each well were obtained at 450 nm and 650 nm, and the data were processed and analyzed using GraphPad Prism software (GraphPad Software Inc). The test results are shown in Table 3.

[0285] Table 3 Prostate cancer cell activity test results of compounds (IC 50 :nM)

[0286] IC 50 :<20nM=*; 20-50nM=**; 50-500nM=***; >500nM=****.

[0287] It can be seen from the data in Table 2 that the novel CDK12 / 13 covalent inhibitor compound containing a cyclic structure of the present invention has a strong inhibitory activity on the proliferation of VCaP cells.

[0288] Example 51 Pharmacokinetic Test

[0289] After a single oral dose (10 mg / kg) and intravenous injection (2 mg / kg) of SD rats, blood samples were collected at appropriate time points. The samples were anticoagulated with EDTA-K2 and centrifuged at 6800×g for 6 minutes at 2-8°C; the supernatant was taken and stored at -80°C for analysis. Plasma sample proteins were precipitated with methanol, vortexed for 1 minute, and then centrifuged at 14000rpm for 7 minutes. The supernatant was used for HPCL-MS analysis. The data were fitted with parameters using DAS2.0 to obtain compartmental model and non-compartmental model parameters. The oral bioavailability of the compound was calculated based on the area under the plasma concentration-time curve (AUC) data. The results are shown in Table 4. The experimental results show that compound ZLC-8-53 has excellent oral pharmacokinetic properties.

[0290] Table 4 Pharmacokinetic experimental results of compounds ZLC-8-53, ZLC-11-74, and ZLC-12-48

[0291] C max Refers to the maximum blood drug concentration, T 1 / 2 is the half-life, CL refers to the clearance, and F refers to the bioavailability.

[0292] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A CDK12 / 13 covalent inhibitor having the structure of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, or a prodrug molecule thereof: in, X and Y are independently selected from the following groups: N or CR5; R5 is selected from the group consisting of hydrogen, halogen, cyano, hydroxy, amino, halomethyl, halomethoxy, haloethyl, haloethoxy, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, and C1-C6 alkyl-substituted amino; Ring A is selected from: Where s is 0, 1, 2 or 3; F, G, H, I, K, L, P, Q, T, U, and Z are independently selected from the following groups: N or CR'; R' is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C1-C6 cycloalkyl, C3-C8 cycloalkyloxy, (CH2)C1-C6 cycloalkyl; J is selected from the group consisting of O, S, NH, NR"; R" is selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl; R1 is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, C3-C8 cycloalkyloxy; W is selected from the group consisting of chemical bonds, R2 is selected from the group consisting of H, -NHR6, -OR6, -CH(R8)R6; R6 is selected from the group consisting of: -(C(R8)R7)R9, -(CH2) n R9; wherein n is selected from: 0, 1 or 2; R7 and R8 are each independently selected from the group consisting of hydrogen, halogen, cyano, methyl, halomethyl, methoxy, halomethoxy, ethyl, haloethyl, ethoxy, haloethoxy, hydroxy, amino, and a 3-8 membered heterocycle containing 1, 2 or 3 heteroatoms; or R7 and R8 are joined together to form a 3-7 membered heterocycle containing 1, 2 or 3 heteroatoms through the carbon atoms to which they are joined; or R7 and R8, together with the carbon atoms to which they are joined, form a substituted or unsubstituted saturated C1-C8 monocyclic, condensed, spirocyclic or bridged ring; R9 is selected from the group consisting of: 1) C1-C8 alkyl, halogenated C1-C4 alkyl, C1-C4 alkoxy, C3-C 10 Cycloalkyl, substituted or unsubstituted 3-8 membered aromatic ring or saturated ring containing O, S or N, 8-12 membered fused ring, spiro ring or bridged ring containing n heteroatoms, n is selected from: 1, 2 or 3, heteroatoms are selected from: O, N, S; 2) A, B, C, D, and E are independently selected from: CH, N, or CR 10 ; R 10 Selected from the group consisting of halogen, cyano, hydroxy, amino, nitro, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C4 alkoxy, halogenated C1-C4 alkoxy, C3-C8 cycloalkyl; A', B', C', D', E' are independently selected from CH, N, NH, S, O, NR 10' or CR 10” R 10' 、R 10” Each independently selected from: C1-C5 alkyl, C1-C5 cycloalkyl; V is N or CR3; Each R3 is independently selected from the following group: H, halogen, cyano, hydroxy, amino, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, halogenated C1-C3 alkoxy, C3-C8 cycloalkyl, -(CH2) m R 11 、-NH(CH2) m R 11 、-NR 14 (CH2) m R 11 、-O(CH2) m R 11 ; a substituted or unsubstituted 3-8 membered heterocyclic ring having 1, 2 or 3 heteroatoms; an 8-12 membered fused, spiro or bridged ring containing 1, 2 or 3 heteroatoms; and at least one R3 is not H; R 11 Selected from the following group: C1-C6 alkyl or NR 12 R 13 ; Among them, R 12 、R 13 are independently selected from: H, C1-C8 alkyl, -(CH2) m NR 14 R 15 、-(CH2) n CR 14 R 15 R 16 , or R 12 、R 13 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted monocyclic, condensed, spirocyclic or bridged ring containing a heteroatom; R 14 、R 15 、R 16 Each independently selected from the following group: H, C1-C8 alkyl, or R 14 、R 15 Together with the nitrogen atom or carbon atom to which they are attached, they form a substituted or unsubstituted monocyclic, condensed, spirocyclic or bridged ring containing 0-3 heteroatoms; m and n are independently selected from: 0, 1, 2, 3, 4, 5, 6, 7 or 8; R4 is selected from the following group: H, R 17 Selected from the group consisting of hydrogen, trifluoromethyl, R 18 Selected from the group consisting of hydrogen, fluorine or methyl; Unless otherwise specified, the term "substituted" refers to the replacement of one or more hydrogen atoms on a group by a substituent selected from the group consisting of halogen, oxo, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C1-C6 acyl, unsubstituted or halogenated C1-C6 amide, unsubstituted or halogenated C1-C6 alkylamino, unsubstituted or halogenated C1-C6 alkyl-hydroxy, unsubstituted or halogenated C3-C6 alkyl, or a 4-8 membered heterocyclyl which is unsubstituted, halogenated or substituted by C1-C4 alkyl; any of the heteroatoms being selected from the group consisting of O, N, and S.

2. The CDK12 / 13 covalent inhibitor 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 independently selected from: N or CR5; R5 is selected from the group consisting of hydrogen, halogen, cyano, halomethyl, and halomethoxy.

3. The CDK12 / 13 covalent inhibitor 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: Where s is 0, 1, 2 or 3; F, G, H, I, K, L, P, Q, U, and Z are independently selected from the following groups: N or CR'; R' is selected from the group consisting of H, cyano, and halogen; J is selected from the group consisting of O, S, NH, NR"; R" is selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl; R1 is selected from the group consisting of H, cyano, halogen, halomethyl, halomethoxy, haloethoxy, haloethyl, C1-C6 alkyl, C3-C8 cycloalkyl, and C1-C6 alkoxy.

4. The CDK12 / 13 covalent inhibitor according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: W is selected from:

5. The CDK12 / 13 covalent inhibitor according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: R2 is selected from the group consisting of: -NHR6; R6 is selected from the group consisting of: -(C(R8)R7)R9; R7 and R8 are each independently selected from the group consisting of hydrogen, halogen, cyano, methyl, halomethyl, methoxy, halomethoxy, ethyl, or R7 and R8 together with the carbon atoms to which they are attached form a saturated C1-C6 monocyclic, spirocyclic or bridged ring; R9 is independently selected from the following group: phenyl, halophenyl, cyano-substituted phenyl, alkoxy-substituted phenyl, pyridyl, pyrimidinyl, isopropyl, tert-butyl, trifluoromethyl, difluoromethyl, cyano, substituted or unsubstituted pyrrolyl, N-methylpyrrolyl, N-methylimidazolyl, N-methylpyrazolyl, imidazolyl, substituted or unsubstituted furanyl, substituted or unsubstituted thienyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted isoxazolyl, substituted or unsubstituted oxazolyl, halo-substituted C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkyl, C3~C7 cycloalkyl, C3~C7 epoxyalkyl.

6. The CDK12 / 13 covalent inhibitor according to claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: Each R3 is independently selected from the following group: H, halogen, cyano, hydroxy, amino, C1-C3 alkyl, halogenated C1-C3 alkyl, C1-C3 alkoxy, halogenated C1-C3 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, 7. The CDK12 / 13 covalent inhibitor according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: R4 is selected from: H, 8. The CDK12 / 13 covalent inhibitor according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: It has the structure shown in formula (II):

9. The CDK12 / 13 covalent inhibitor according to claims 1-8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: The compound is selected from:

10. Use of the CDK12 / 13 covalent inhibitor or its pharmaceutical composition according to any one of claims 1 to 9, or its pharmaceutically acceptable salt, or its stereoisomer or its prodrug molecule in the preparation of a medicament for preventing and / or treating diseases mediated by CDK12 / 13 serine / threonine protein kinase.

11. The use according to claim 10, characterized in that The disease mediated by CDK12 / 13 serine / threonine protein kinase is selected from the following group: prostate cancer, breast cancer, uterine cancer, ovarian cancer, non-small cell lung cancer, small cell lung cancer, Ewing 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.

12. A pharmaceutical composition for preventing and / or treating tumors, characterized in that: The invention comprises an active ingredient and a pharmaceutically acceptable excipient, wherein the active ingredient comprises a novel CDK12 / 13 covalent inhibitor substituted with a ring structure according to any one of claims 1 to 9, or a pharmaceutical composition thereof, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof.

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