Aryl amide compounds and uses thereof

Abstract

The present application relates to a kind of arylamide compounds or its pharmaceutically acceptable salt or its stereoisomer with the structure shown in formula I, formula II, formula III or formula IV, and its application.The arylamide compound provided by the present application is a molecular glue degrader for Bcr-Abl T315I , which can effectively degrade wild-type Bcr-Abl kinase and mutant Bcr-Abl T315I Kinase, can effectively inhibit the kinase activity of Bcr-Abl and Bcr-Abl T315I Mutant, has good anti-proliferation effect on tumor cells carrying Bcr-Abl and Bcr-Abl T315I ;And selectivity is good, side effect is small, and safety is good.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to antitumor drugs, and more specifically to arylamide compounds and their role as Bcr-Abl. T315I Applications of molecular adhesives. Background Technology

[0002] Chronic myeloid leukemia (CML) is a malignant clonal disease of the bone marrow hematopoietic stem cells in the blood system, ranking third in incidence among leukemias. The cause of CML is a mutation of the Philadelphia chromosome (Ph chromosome), specifically the recombination of the c-Abl proto-oncogene on chromosome 9 with the BCR gene on chromosome 22 to form the Bcr-Abl fusion gene. This fusion gene causes abnormal activation of tyrosine kinases, leading to abnormal cell proliferation and the occurrence and progression of CML.

[0003] Tyrosine kinase inhibitors (TKIs) are currently the mainstream treatment for chronic myeloid leukemia (CML), exerting their therapeutic effect by precisely inhibiting the activity of the oncogenic protein Bcr-Abl. These drugs can significantly prolong patient survival and help many patients achieve improved blood parameters or even a cancer-free state. Imatinib, as the first approved Bcr-Abl targeted therapy, has been the gold standard of treatment since its launch in 2001. Clinical data shows that it enables more than 80% of patients to achieve long-term survival, greatly advancing the progress of targeted cancer therapy. However, with increased clinical use, approximately 30% of patients develop resistance to its treatment. Among these, the T315I mutation is the most common mutation, accounting for approximately 13%–16% of mutations. This mutation weakens the efficacy of all generations of TKIs, from imatinib and dasatinib to the third-generation ponatinib. Although ponatinib maintains activity against the T315I mutation, its severe captopril side effects and inability to eliminate the Bcr-Abl scaffold protein result in a 5-year relapse rate as high as 60%. Continuous scaffold protein reactivation of downstream signaling becomes a molecular reservoir for disease relapse. It is evident that small molecule inhibitors targeting kinases cannot avoid secondary mutations during clinical use, leading to clinical resistance and consequently affecting efficacy. Therefore, research is needed on substances capable of degrading Bcr-Abl. T315I Inhibitors of function have important clinical significance.

[0004] Targeted protein degradation (TPD) technology utilizes the naturally occurring protein degradation systems within human cells to induce selective ubiquitination and degradation of target proteins, thereby transiently eliminating the biological function of the target protein and reducing its activity at its source. Compared to traditional inhibitors or agonists, it can significantly reduce the incidence of drug resistance mutations. Among these technologies, protein degradation-targeted chimeras (PROTACs) and molecular glue degraders have received the most widespread attention.

[0005] Protein degradation-targeting chimeras (PROTACs) facilitate the recognition of target proteins by the proteasome and their degradation via the intracellular ubiquitin-proteasome system (UPS). PROTACs are heterobifunctional compounds consisting of two independent ligands linked by a chemical linker; one ligand binds to the target protein, and the other binds to an E3 ubiquitin ligase. This structural design allows PROTACs to bring the target protein and the E3 ligase closer together, forming a ternary complex that promotes ubiquitination and subsequent proteasome degradation of the target protein. Theoretically, PROTACs designed for the T315I mutant Bcr-Abl fusion protein could eliminate Bcr-Abl... T315I Enzymatic activity and scaffold function. Lu Xiaoyun et al. designed and obtained a representative degrader of PROTACs, 7o, based on Olverembatinib (GZD824) linked to a cereblon (CRBN) ligand. This degrader exhibited good enzyme activity and scaffold function against Bcr-Abl at a concentration of 100 nM. T315I It exhibited significant degradation effects, and in Ba / F3Bcr-Abl T315I The cells exhibited activity at a half-maximal inhibitory concentration (HMC) of 26.8 nM. Subsequently, other PROTACs designed based on imatinib, dasatinib, and pannatinib were also reported to inhibit Bcr-Abl or Bcr-Abl. T315I They exhibit significant degradation effects. However, the high molecular weight (>1kDa), poor membrane permeability, and complex ternary complex formation kinetics of these PROTACs constitute important obstacles to clinical translation.

[0006]

[0007] Compared to protein degradation-targeting chimeras, molecular glue degraders are low-molecular-weight compounds that promote target protein degradation by enhancing the interaction between the target protein and the E3 ligase. Unlike PROTACs, molecular glue degraders typically do not directly bind to the target protein or E3 ligase, but rather enhance their binding affinity to the E3 ligase by binding to the target protein's interactors or altering the target protein's conformation. This mechanism is similar to a "molecular glue," enhancing the surface complementarity and intrinsic binding affinity between two proteins, thereby promoting ubiquitination and subsequent target protein degradation induced by protein-ligase interactions. Molecular glue degraders generally have smaller molecular weights than PROTACs, and through proper design, they can possess not only the excellent properties of small molecule drugs but also potent degradation effects. However, current research on Bcr-Abl... T315I There are very few research reports on molecular adhesive degraders. Summary of the Invention

[0008] Based on this, the purpose of this invention is to provide a solution for Bcr-Abl T315I A molecular adhesive degrader that can effectively degrade Bcr-Abl T315I Kinase, thereby effectively inhibiting Bcr-Abl T315I The kinase activity of the mutant, for those carrying Bcr-Abl T315I It has a good anti-proliferative effect on tumor cells.

[0009] The technical solutions for achieving the above objectives include the following.

[0010] In a first aspect, the present invention provides an aryl amide compound having the structure shown in Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0011]

[0012] Among them, R1, R2, R3, and R'3 are independently selected from: C2-C8 alkyl, cyano, benzyl, ...

[0013] R4 is selected from: hydrogen, cyano, and halogen;

[0014] R5 is selected from: hydrogen, One or more R9-substituted or unsubstituted C1-C6 alkyl groups, one or more R9-substituted or unsubstituted C3-C6 cycloalkyl groups, one or more R9-substituted or unsubstituted C6-C6 cycloalkyl groups. 10 aryl; R8 is selected from: one or more R9-substituted or unsubstituted C6-C 10 Aryl, C1-C6 alkoxy; each R9 is independently selected from: hydrogen, C1-C6 alkylamino, (C1-C6 alkyl)2amino, halogen, halogen-substituted C1-C6 alkyl;

[0015] R6 is selected from: hydrogen, one or more R 10 Substituted or unsubstituted C1-C8 alkyl, C6-C 10 Aryl, 3-8 membered heterocyclic groups, Each R 10 R independently 10 Selected from: hydrogen, halogen, azide, C1-C6 alkyl acyl, formaldehyde, amino, hydroxyl, cyano, cyclopropyl;

[0016] R7 is selected from: C1-C6 alkyl groups;

[0017] R is selected from: hydrogen, C1-C6 alkyl, C1-C6 alkoxy.

[0018] In some embodiments, R4 is selected from: hydrogen, cyano, fluorine, and chlorine.

[0019] In some embodiments, R4 is selected from: hydrogen, cyano, and fluorine;

[0020] In some embodiments, R5 is selected from: hydrogen, One or more R9-substituted or unsubstituted C1-C3 alkyl, cyclopropyl, one or more R9-substituted or unsubstituted phenyl; R8 is selected from: one or more R9-substituted or unsubstituted phenyl, C1-C3 alkoxy; each R9 is independently selected from: hydrogen, C1-C3 alkylamino, (C1-C3 alkyl)2amino, fluorine, chlorine, bromine, trifluoromethyl.

[0021] In some implementations, R5 is selected from: Hydrogen, cyclopropyl Trifluoromethyl

[0022] In some embodiments, R6 is selected from: hydrogen, C1-C6 alkyl, one or more R 10 Substituted C1-C3 alkyl, phenyl, Each R 10 Each group is independently selected from: fluorine, chlorine, bromine, azide, C1-C3 alkyl acyl, formaldehyde, amino, hydroxyl, cyano, and cyclopropyl.

[0023] In some implementations, R6 is selected from: Dichloromethyl, bromochloromethyl, dibromomethyl, chlorofluoromethyl, difluoromethyl, phenyl, methyl, hydrogen, ethyl, chloromethyl, bromomethyl, 2-azidoethyl, 2-acetylethyl, 2-formaldehydeethyl, 2-aminoethyl, 2-hydroxyethyl, n-butyl, n-hexyl.

[0024] In some embodiments, R7 is selected from C1-C3 alkyl groups, preferably methyl.

[0025] In some implementations, R1 is selected from:

[0026]

[0027] R2 is selected from:

[0028]

[0029] R3 is selected from: R'3 is selected from: In some embodiments, R is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy.

[0030] In some embodiments, R is selected from: hydrogen, methyl, ethyl, methoxy, ethoxy. In some embodiments, R is selected from: hydrogen, methyl.

[0031] In some embodiments, the arylamide compound is selected from the following compounds:

[0032]

[0033]

[0034] Secondly, the present invention provides the use of the aryl amide compounds or their pharmaceutically acceptable salts or stereoisomers in the preparation of Bcr-Abl kinase inhibitors.

[0035] Thirdly, the present invention provides the use of the arylamide compounds or their pharmaceutically acceptable salts or stereoisomers in the preparation of mutant Bcr-Abl kinase inhibitors; preferably, the mutations include: T315I mutation, M244V mutation, G250E mutation, Q252H mutation, H396P mutation, Y253F mutation, E255K mutation, F317L mutation; more preferably, the mutation is T315I mutation.

[0036] Fourthly, the present invention provides the use of the aryl amide compounds or their pharmaceutically acceptable salts or stereoisomers in the preparation of Bcr-Abl protein degrading agents.

[0037] Fifthly, the present invention provides the use of the aryl amide compounds or their pharmaceutically acceptable salts or stereoisomers in the preparation of mutant Bcr-Abl protein degraders; preferably, the mutations include: T315I mutation, M244V mutation, G250E mutation, Q252H mutation, H396P mutation, Y253F mutation, E255K mutation, and F317L mutation; more preferably, the mutation is the T315I mutation.

[0038] In a sixth aspect, the present invention provides the use of the aforementioned aryl amide compounds or pharmaceutically acceptable salts thereof or their stereoisomers in the preparation of medicaments for the treatment and / or prevention of tumors.

[0039] In some embodiments, the tumor is a tumor carrying the Bcr-Abl fusion gene.

[0040] In some embodiments, the tumor is a tumor carrying a mutated Bcr-Abl gene; preferably, the mutation includes: T315I mutation, M244V mutation, G250E mutation, Q252H mutation, H396P mutation, Y253F mutation, E255K mutation, F317L mutation; more preferably, the mutation is T315I mutation.

[0041] In some embodiments, the tumor is leukemia, colorectal adenocarcinoma, breast cancer, or pancreatic cancer, preferably leukemia. The leukemia is preferably acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, or acute lymphoblastic leukemia. More preferably, the tumor cells of the leukemia are MV4-11 cells or K562 cells.

[0042] In a seventh aspect, the present invention provides a pharmaceutical composition for the prevention or treatment of tumors, prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises an arylamide compound as described in the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0043] The present invention has the following beneficial effects:

[0044] The aryl amide compound provided by this invention is a target for Bcr-Abl T315I It is a molecular glue degrader of kinases, which can effectively degrade wild-type Bcr-Abl kinase and Bcr-Abl T315I Mutant kinases can effectively inhibit Bcr-Abl and Bcr-Abl T315I The kinase activity of mutants, for those carrying Bcr-Abl and Bcr-Abl T315I Tumor cells with mutant genes have good anti-proliferative effects; and they are highly selective, have few toxic side effects, and are safe.

[0045] The aryl amide compounds of this invention can induce Bcr-Abl and Bcr-Abl using the protein degradation system naturally present in human cells. T315I Selective ubiquitination and degradation of the protein can transiently eliminate Bcr-Abl and Bcr-Abl. T315I The biological function of the protein can reduce Bcr-Abl and Bcr-Abl at their source. T315I Compared with traditional inhibitors, this method can significantly reduce the incidence of drug resistance mutations by improving protein activity.

[0046] The further preferred compounds of this invention possess excellent pharmacokinetic properties, exhibiting good oral absorption efficiency and bioavailability. Their high bioavailability and low pharmacokinetic variability (SD < 15%) provide a solid foundation for druggability and have the potential to treat Bcr-Abl and Bcr-Abl carriers. T315I These are candidate oral drugs for tumors and have potential for clinical application. Attached Figure Description

[0047] Figure 1 For the compound's effect on the target protein Bcr-Abl T315I Immunoblot image.

[0048] Figure 2 The degradation activities of compounds HTY-4147 and HTY-5018 on BCR-ABL protein in K562 cells were investigated.

[0049] Figure 3 These are the pharmacokinetic data for compound HTY-4147.

[0050] Figure 4 The study investigated the inhibitory effect of compound HTY-4147 on the proliferation of BaF3-BCR-ABL-T315I xenografts in vivo.

[0051] Figure 5 The study investigated the inhibitory effects of compound HTY-4147 on the cell cycle and apoptosis of BaF3-BCR-ABL-T315I.

[0052] For convenience, the letters "HTY-" or "GZD" are omitted from the compound numbers in some figures. For example, 4147 in the figure corresponds to HTY-4147, 4101 corresponds to HTY-4101, and 824 in the figure corresponds to GZD824. Detailed Implementation

[0053] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0054] Experimental methods in the following examples, unless otherwise specified, were performed under conventional conditions, such as those described in the fourth edition of *Molecular Cloning: A Laboratory Manual*, edited by Green and Sambrook, published in 2013, or as recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0055] Furthermore, as used herein, the term "or" is an inclusive "or" sign and is equivalent to the term "and / or" unless the context clearly specifies otherwise. The term "based on" is not exclusive and allows for basing on other factors not described unless the context clearly specifies otherwise. Additionally, throughout the specification, the meanings of "an," "a," and "the" include plural indicators. The meaning of "in" includes both "in" and "on."

[0056] In the compounds of this invention, when any variable (e.g., R9, etc.) appears more than once in any component, the definition of each occurrence is independent of the definitions of other occurrences. Similarly, combinations of substituents and variables are permitted, provided such combinations stabilize the compound. A line drawn from a substituent into the ring system indicates that the bond referred to can be attached to any substituted ring atom. If the ring system is polycyclic, it means that such a bond is attached only to any suitable carbon atom of a neighboring ring. It will be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of this invention to provide chemically stable compounds that can be easily synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is itself substituted by more than one group, it should be understood that these groups can be on the same carbon atom or different carbon atoms, as long as structural stability is achieved.

[0057] As used herein, the term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms. For example, the definition of "C1-C6" in "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0058] As used herein, the term "cycloalkyl" refers to a monocyclic, bicyclic, or polycyclic cyclic hydrocarbon group whose ring atoms are composed of carbon atoms and are saturated or partially unsaturated. Bicyclic or polycyclic groups include spirocyclic, fused, and bridged rings. For example, "cycloalkyl" includes, but is not limited to, the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. wait.

[0059] The term "alkoxy" as used in this article refers to a group having an -O-alkyl structure, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0060] As used herein, the terms "heterocyclic alkyl" or "heterocyclic group" refer to saturated or partially unsaturated monocyclic, bicyclic, or polycyclic cyclic substituents (including spirocyclic, bridged, fused, and fused rings, etc.), wherein one or more ring atoms are selected from N, O, or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. Examples include: morpholinyl, piperidinyl, tetrahydropyrrolyl, pyrrolylalkyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazoleyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothiophene, dihydrotriazolyl, dihydroazacyclobutane, tetrahydrofuranyl, tetrahydrothiophene, etc. And so on, and their N-oxides. The connection of heterocyclic substituents can be achieved through carbon atoms or through heteroatoms.

[0061] As will be understood by those skilled in the art, the term “halo” or “halogen” as used herein refers to chlorine, fluorine, bromine, and iodine.

[0062] This invention includes the free forms of compounds of formulas I-IV, as well as their pharmaceutically acceptable salts and stereoisomers. Some specific exemplary compounds described herein are amine compounds. The term "free form" refers to an amine compound in its non-salt form. Pharmaceutically acceptable salts included include not only exemplary salts of the specific compounds described herein, but also typical pharmaceutically acceptable salts of the free forms of all compounds of formulas I-IV. The free forms of specific salts of said compounds can be separated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a suitable dilute aqueous solution of an alkali, such as dilute aqueous solution of NaOH, potassium carbonate, dilute ammonia, or sodium bicarbonate. The free form may differ somewhat from its respective salt form in certain physical properties, such as solubility in polar solvents, but for the purposes of this invention, such acid salts and alkali salts are otherwise pharmaceutically equivalent to their respective free forms.

[0063] Pharmaceutically acceptable salts of the present invention can be synthesized from compounds of the present invention containing either a basic or acidic moiety using conventional chemical methods. Typically, salts of basic compounds are prepared by ion-exchange chromatography or by reacting a free base with a stoichiometric or excess amount of the desired salt form of an inorganic or organic acid in a suitable solvent or a combination of solvents. Similarly, salts of acidic compounds are formed by reacting with a suitable inorganic or organic base.

[0064] Therefore, pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the present invention formed by reacting an alkaline compound of the present invention with an inorganic or organic acid. For example, conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc., and also include 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, pyric acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethylsulfonic acid, trifluoroacetic acid, etc.

[0065] If the compounds of this invention are acidic, then a suitable "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic alkali, including inorganic and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganese salts, potassium salts, sodium salts, zinc salts, etc. Ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases, including salts of primary, secondary, and tertiary amines, wherein substituted amines include 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, hydroxycobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, guanidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, aminobutanetriol, etc.

[0066] Since the deprotonated acidic portion of the compound, such as the carboxyl group, can be anionic under physiological conditions, and this charge can then be balanced by the protonated or alkylated basic portion, such as the tetravalent nitrogen atom, which carries a cation, it should be noted that the compounds of the present invention are potential internal salts or zwitterions.

[0067] The pharmaceutical composition or method for the prevention and / or treatment of tumors provided by this invention comprises (administered to a patient or subject) an active ingredient (i.e., the aryl amide compound described in this invention or its pharmaceutically acceptable salt or stereoisomer) within a safe and effective range, as well as pharmaceutically acceptable excipients. When administering the drug, a safe and effective amount of the active ingredient is applied to the mammal (such as a human) requiring treatment, wherein the dose administered is a pharmaceutically considered effective dose. Of course, the specific dose should also consider factors such as the route of administration and the patient's health condition, which are all within the scope of a skilled physician's expertise.

[0068] The "active ingredient" referred to in this invention refers to the compounds of formulas I-IV described in this invention, or their pharmaceutically acceptable salts or their stereoisomers.

[0069] The "active ingredient" and pharmaceutical composition described in this invention can be used to prepare drugs for the prevention and / or treatment of tumors.

[0070] "Safe and effective dose" means that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects.

[0071] "Pharmaceutical acceptable excipients" refer to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0072] "Compatibility" here refers to the ability of the components in the composition to interact with and blend with the active ingredients of the present invention without significantly reducing the efficacy of the active ingredients.

[0073] Pharmaceutically acceptable examples of excipients include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (such as...). Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0074] In another preferred embodiment, the compounds of formulas I-IV of the present invention can form complexes with macromolecular compounds or polymers through non-bonding interactions. In another preferred embodiment, the compounds of formulas I-IV of the present invention, as small molecules, can also be linked to macromolecular compounds or polymers through chemical bonds. The macromolecular compounds can be biological macromolecules such as polysaccharides, proteins, nucleic acids, polypeptides, etc.

[0075] There are no particular limitations on the administration of the active ingredients or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0076] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0077] In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components:

[0078] (a) Fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol and silica;

[0079] (b) Adhesives, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0080] (c) Moisturizers, such as glycerin;

[0081] (d) Disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginate, certain complex silicates, and sodium carbonate;

[0082] (e) Slow solvents, such as paraffin;

[0083] (f) Absorption accelerators, such as quaternary ammonium compounds;

[0084] (g) Wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0085] (h) Adsorbents, such as kaolin; and

[0086] (i) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain a buffer.

[0087] The solid dosage form can also be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active ingredient from this composition can be delayed in a portion of the digestive tract. Examples of suitable encapsulating components are polymers and waxes.

[0088] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0089] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0090] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof. The invention is further described in detail below with reference to specific examples. The compounds prepared in the following examples are:

[0091]

[0092] in,

[0093]

[0094] In the following examples, the abbreviations for each reagent or raw material are as follows: NBS: N-bromosuccinimide

[0095] AIBN: Azobisisobutyronitrile

[0096] DCM: Dichloromethane

[0097] HATU: 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate

[0098] DMF: N,N-Dimethylformamide

[0099] DIPEA: N,N-Diisopropylethylamine

[0100] DDQ: 2,3-Dichloro-5,6-dicyanobenzoquinone

[0101] THF: Tetrahydrofuran

[0102] Example 1

[0103]

[0104] In the first step, 2.0 g (9.75 mmol) of compound 2-methyl-5-nitrotrifluorotoluene, 2.08 g (11.7 mmol) of NBS, and 160 mg (0.975 mmol) of AIBN were weighed into a reaction flask, and 20 mL of carbon tetrachloride was added. The reaction was carried out overnight at 95 °C. After the reaction was completed, the solvent was evaporated under vacuum, and the compound was purified by silica gel column chromatography (100% PE) to obtain compound 2 (744 mg, yield 26.97%). 1 H NMR (400MHz, DMSO-d6) δ8.54(dd,J=8.5,2.5Hz,1H),8.43(d,J=2.4Hz,1H),8.05(d,J=8.5Hz,1H),4.88(s,2H).

[0105] In the second step, compound 2 (350 mg, 1.23 mmol) was dissolved in DCM, followed by the addition of triethylamine (0.25 mL, 1.84 mmol) and tert-butylpiperazine carboxylate (253 mg, 1.36 mmol), and the reaction was carried out at room temperature for 5 hours. After the reaction was completed, the solvent was evaporated under vacuum, and the compound was purified by silica gel column chromatography (PE:EA = 25:1 to 10:1) to obtain compound 3 (429 mg, yield 89.62%). 1 H NMR (400MHz, DMSO-d6) δ8.51 (dd, J=8.6, 2.4Hz, 1H), 8.41 (d, J=2.4Hz, 1H), 8.11 (d,J=8.6Hz,1H),3.75(s,2H),3.35(s,4H),2.38(t,J=5.1Hz,4H),1.39(s,9H).

[0106] In the third step, compound 3 (1.44 g, 3.7 mmol) was dissolved in a mixed solvent of EtOH and H2O (2:1), and iron powder (1.03 g, 18.5 mmol) and ammonium chloride (594 mg, 11.1 mmol) were added. The mixture was reacted overnight at 60 °C. After the reaction was completed, the mixture was filtered, the filter cake was washed with ethanol, the filtrate was collected, concentrated, and the organic phase was washed with saturated sodium chloride solution. The aqueous phase was extracted three times with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The mixture was purified by silica gel column chromatography (PE / EA = 10:1 to 3:1) to give a white solid compound 4 (1.01 g, yield 76.03%). 1H NMR (400MHz, DMSO-d6) δ7.30(d,J=8.3Hz,1H),6.86(s,1H),6.75(d,J=8.2Hz,1H),5.45(s,2H),3.40(s,2H),3.29(s,4H),2.27(s,4H),1.38(s,9H).

[0107] In the fourth step, methyl 3-iodo-4-methylbenzoate (1.0 g, 3.62 mmol), palladium dichloride bis(triphenylphosphine) (25 mg, 0.036 mmol), and cuprous iodide (6.8 mg, 0.036 mmol) were weighed into a reaction flask. Under argon protection, triethylamine (3.17 mL, 22.91 mmol), trimethylsilylacetylene (2.54 mL, 18 mmol), and acetonitrile (6 mL) were added, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, the mixture was filtered, the filter cake was washed with acetonitrile, and the concentrated filtrate was collected to obtain compound 6. This compound was then redissolved in 6 mL of methanol, and potassium carbonate (1.0 g, 7.23 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the mixture was filtered, the filter cake was washed three times with methanol, and the concentrated filtrate was collected and purified by silica gel column chromatography (100% PE to PE:EA = 50:1) to obtain compound 7 (539 mg, yield 85.91%). 1 H NMR (400MHz, DMSO-d6) δ7.94(s,1H),7.85(d,J=9.1Hz,1H),7.46(d,J=8.8Hz,1H),4.54-4.48(m,1H),3.84(s,3H),2.45(s,3H).

[0108] In step 5, compound 7 (730 mg, 4.19 mmol), 5-bromo-1H-pyrazolopyridine (830 mg, 4.19 mmol), palladium dichloride bis(triphenylphosphine) (147 mg, 0.209 mmol), and cuprous iodide (79 mg, 0.419 mmol) were weighed into a reaction flask. Under argon protection, triethylamine (4 mL, 29 mmol) and acetonitrile (10 mL) were added, and the reaction was carried out overnight at 82 °C. After the reaction was completed, the mixture was filtered, and the filter cake was washed three times with acetonitrile. No purification was required, and the mixture was used directly in the next step to obtain compound 8 (507 mg, yield 42.25%). 1 H NMR (400MHz, CDCl3) δ11.98(s,1H),8.78(d,J=1.9Hz,1H),8.30(d,J=1.9Hz,1H),8.21(d,J=1.8H z,1H),8.15(s,1H),7.92(dd,J=8.0,1.9Hz,1H),7.34(d,J=8.0Hz,1H),3.93(s,3H),2.60(s,3H).

[0109] Step 6: Compound 4 (718 mg, 2.0 mmol) and Compound 8 (582 mg, 2.0 mmol) were weighed into a reaction flask. Under argon protection, anhydrous tetrahydrofuran (10 mL) was added. Then, at -78 °C, a 1 M potassium tert-butoxide tetrahydrofuran solution (12 mL, 12.0 mmol) was added. After reacting for 1 hour, the mixture was brought to room temperature, water was added, and the aqueous phase was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was evaporated under vacuum, and the mixture was purified by silica gel column chromatography (PE:EA = 5:1 to 1:1) to obtain Compound 9 (1.01 g, yield 80.90%). 1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),10.55(s,1H),8.74(s,1H),8.53(s,1H),8.22(s,2H),8.19(s,1H),8.08(d,J=8.7Hz, 1H),7.92(d,J=8.0Hz,1H),7.73(d,J=8.6Hz,1H),7.53(d,J=7.4Hz,1H),3.59(s,2H),2.59(s,3H),2.34(s,4H),1.39(s,9H).

[0110] Step 7: Weigh compound 9 (500 mg, 0.8 mmol) into a reaction flask, add 5 mL of 1,4-dioxane solution in 4 M hydrochloric acid, and react at room temperature for 4 hours. After the reaction is complete, evaporate the solvent under vacuum, and use it directly in the next step without purification to obtain compound 10 (400 mg, 95.46%). 1 H NMR (400MHz, DMSO-d6) δ10.84(s,1H),9.84(s,2H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H),8.40(d,J=2.1Hz,1H), 8.24-8.21(m,3H),7.97(dd,J=8.0,2.0Hz,1H),7.53(d,J=8.1Hz,1H),4.45(s,2H),3.46(d,J=25.1Hz,9H),2.59(s,3H). 13C NMR(151MHz,DMSO-d6)δ165.5,151.5,151.0,144.5,141.0,134.7,134.2,133.5,132.4,131.2,130.5,129.5,129.3,12 8.8,125.1,123.9,123.3,122.7,117.9,117.8,114.5,112.2,92.5,88.7,66.8,55.6,48.6,20.9.HRMS(ESI)calcd.forC 28 H 25 F3N6O 519.2115[M+H] + ,found 519.2101.

[0111] Step 8: Weigh (E)-4-(4-methoxyphenyl)-4-oxo-2-butenoic acid (34 mg, 0.165 mmol) and HATU (68 mg, 0.18 mmol) and dissolve them in DMF (3 mL). Add DIPEA (74 μL, 0.45 mmol), stir for 5 minutes, then add compound 10 (78 mg, 0.15 mmol), and react overnight at room temperature. After the reaction is complete, add water, extract three times with ethyl acetate, wash the organic layer with brine, and dry with anhydrous sodium sulfate. Evaporate the solvent under vacuum, and purify by silica gel column chromatography (DCM:MeOH = 100:1 to 40:1) to obtain a yellow solid Bal-6 (32 mg, yield 41.78%). 1 HNMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(s,1H),8.53(s,1H),8.21(d,J=13.9Hz,3H),8.07(d,J=25.3Hz,3H),7.93( s,1H),7.78(d,J=16.7Hz,2H),7.53(s,1H),7.41(d,J=14.5Hz,1H),7.09(s,2H),3.86(s,3H),3.62(s,6H),2.59(s,3H),2.44(s,4H). 13C NMR(151MHz,DMSO-d6)δ188.0,165.2,164.2,163.9,151.5,151.0,144.2,13 8.8,134.3,133.9,133.5,133.1,132.6,132.0,131.8,131.6,131.0,130.4, 129.9,128.6,127.9,125.7,124.0,122.7,117.7,114.8,114.5,112.3,92.4 ,88.8,57.7,56.1,53.5,52.8,46.0,42.2,40.5,20.9.HRMS(ESI)calcd.for C 39 H 33 F3N6O4707.2588[M+H] + ,found 707.2587.

[0112] Example 2

[0113]

[0114] Following a similar synthetic method as step eight in Example 1, compound Bal-7 was obtained by reacting 2-cyano-3-cyclopropylacrylic acid and compound 10 as a gray-black solid (35 mg, yield 40.58%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=1.9Hz,1H),8 .23(t,J=2.2Hz,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.93(dd,J=8.1,2.0Hz,1H) ,7.75(d,J=8.6Hz,1H),7.53(d,J=8.1Hz,1H),6.59(d,J=11.1Hz,1H),3.62(s,2H),3.58-3.47(m,4 H),2.59(s,3H),2.43(t,J=4.9Hz,4H),1.88(m,1H),0.92(dd,J=6.6,2.4Hz,2H),0.86-0.81(m,2H). 13C NMR(151MHz,DMSO-d6)δ165.2,164.2,162.0,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.0,131.8,131.0,130.4,128.6,128.0,125 .7,124.0,122.7,117.7,116.2,114.5,112.3,106.6,92.4,88.7,57.7,52.8,34.7,31.8,30.8,29.4,20.9,15.8,10.4.HRMS(ESI)calcd.forC 35 H 30 F3N7O2638.2486[M+H] + ,found 638.2486.

[0115] Example 3

[0116]

[0117] Following a similar synthetic method as step eight in Example 1, compound Bal-8 was obtained by reacting monomethyl fumarate and compound 10 as a yellow solid (30 mg, yield 39.93%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),10.56(s,1H),8.74(s,1H),8.53(s,1H),8 .23(s,2H),8.20(s,1H),8.10(d,J=8.5Hz,1H),7.93(d,J=8.2Hz,1H),7.75(d,J= 8.6Hz,1H),7.53(d,J=8.2Hz,1H),7.47(d,J=15.5Hz,1H),6.57(d,J=15.4Hz,1H) ,3.73(d,J=3.7Hz,3H),3.62(s,2H),3.57(s,4H),2.59(s,3H),2.45-2.37(m,4H). 13C NMR(151MHz,DMSO-d6)δ166.0,165.2,163.1,151.5,150.9,144.2,138.8,135.3,134.3,133.5,132.6,132.0,131.8,131.0,130.4,130 .0,129.2,128.6,127.8,124.0,122.6,117.7,114.5,112.3,92.4,88.8,57.6,53.4,52.7,52.5,46.0,42.1,20.9.HRMS(ESI)calcd.for C 33 H 29 F3N6O4631.2275[M+H] + ,found 631.2274.

[0118] Example 4

[0119]

[0120] Following a similar synthetic method as step eight in Example 1, compound Bal-9 was obtained by reacting (E)-4-(dimethylamino)but-2-enoic acid with compound 10 as a yellow solid (20 mg, yield 23.54%), using an elution system (DCM:MeOH = 100:1 to 20:1). 1 H NMR (400MHz, DMSO-d6) δ13.95(s,1H),10.56(s,1H),8.74(s,1H),8.53(s,1H),8.21(d,J=11.9Hz,3H),8.09(s,1H),7.93(s ,1H),7.75(s,1H),7.53(s,1H),6.59(s,2H),3.60(s,2H),3.55(s,4H),3.04(s,2H),2.59(s,3H),2.38(s,4H),2.15(s,6H). 13 C NMR(151MHz,DMSO-d6)δ165.2,164.4,151.5,151.0,144.2,142.2,138.8,134.2,133.5,132.6,132.1,131.8,131.0,130.4,128.6,128.0 ,125.7,124.0,122.9,122.7,117.8,114.5,112.3,92.4,88.8,60.3,57.7,53.6,52.9,45.6,45.4,41.9,29.5,20.9.HRMS(ESI)calcd.for C 34 H 34F3N7O2630.2799[M+H] + ,found 630.2792.

[0121] Example 5

[0122]

[0123] Following a similar synthetic method as step eight of Example 1, compound Bal-10 was obtained by reacting 3-azidopropionic acid with compound 10 as a white solid (20 mg, yield 26.63%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),10.56(s,1H),8.74(s,1H),8.53(s,1H),8.22(s,3H),8.10(s,1H),7.93( s,1H),7.75(s,1H),7.53(s,1H),3.60(s,2H),3.56(s,4H),2.59(s,3H),2.39(s,4H),1.30(s,2H),1.17(s,2H). 13 C NMR(151MHz,DMSO-d6)δ165.2,164.6,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,127 .8,125.7,124.0,122.7,117.7,114.5,112.3,92.4,88.8,60.2,57.7,53.6,52.9,45.5,42.0,29.4,20.9.HRMS(ESI)calcd.for C 31 H 28 F3N9O2638.2210[M+Na] + ,found 638.2202.

[0124] Example 6

[0125]

[0126] In the first step, K₂CO₃ (3.1 g, 22.53 mmol) was added to an acetone solution of 5-hydroxyindole (1 g, 7.51 mmol) and tert-butyl bromoacetate (1.6 mL, 1.5 mmol), and the mixture was refluxed overnight. After the reaction was completed, the filtrate was collected by filtration, the solvent was evaporated under vacuum, and the solution was purified by silica gel column chromatography (PE:EA = 9:1 to 8:1) to give compound 6-2 (1.78 g, 96% yield).

[0127] In the second step, compound 6-2 (1.78 mg, 7.2 mmol) was dissolved in acetic acid. NaBH3CN (1.35 g, 21.6 mmol) was added in portions to the reaction solution at 10 °C, and the reaction was stirred at this temperature for 4 hours, followed by quenching with water. All volatile organic compounds were removed by vacuum evaporation, and the aqueous layer was extracted three times with dichloromethane. The dichloromethane layer was washed with dilute NaOH solution, then with saturated brine, dried over anhydrous Na2SO4, and the volatile organic solvent was evaporated under reduced pressure. The solution was purified by silica gel chromatography (PE:EA = 4:1 to 3:1) to give compound 6-3 (1.58 g, 87% yield).

[0128] In the third step, K₂CO₃ (1.75 g, 12.72 mmol) was added to an anhydrous THF solution of compound 6-3 (1.58 g, 6.36 mmol). Acryloyl chloride (556 μL, 7.0 mmol) was added dropwise to the reaction solution at 0 °C with vigorous stirring. The reaction was stirred at this temperature for 30 minutes, then quenched with water. All volatile organic compounds were removed by evaporation under reduced pressure, and the aqueous layer was extracted with ethyl acetate. The ethyl acetate layer was then washed with saturated brine and dried over anhydrous Na₂SO₄. The volatile organic solvent was evaporated under reduced pressure, and the solution was purified by silica gel chromatography (PE:EA = 8:1 to 2:1) to give compound 6-4 (1.08 g, 56% yield).

[0129] In the fourth step, DDQ (1.1 g, 4.85 mmol) was added to an anhydrous toluene solution of compound 6-4 (1.09 g, 3.59 mmol), and the mixture was heated under reflux overnight. After the reaction was complete, the reaction mixture was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous Na₂SO₄, and the volatile organic solvent was evaporated under reduced pressure. The crude product was purified by silica gel chromatography (PE:EA = 5:1) to give compound 6-5 (864 mg, 80% yield).

[0130] In step 5, 85% H3PO4 (4 mL) was added to an acetonitrile solution of compound 6-5 (400 mg, 1.33 mmol), and the mixture was stirred overnight at room temperature. The reaction was then quenched with water, and the volatile organic compounds were removed by evaporation under reduced pressure. The aqueous layer was extracted twice with ethyl acetate and washed with saturated brine. The mixture was dried over anhydrous Na2SO4, and the volatile organic solvent was evaporated under reduced pressure. The crude product was purified by silica gel chromatography (DCM:MeOH = 10:1) to give compound 6-6 (244 mg, 75% yield).

[0131] Step 6: Following a similar synthetic method to step 8 of Example 1, compound 6-6 and compound 10 were reacted to obtain compound Bal-10, which was a white solid (26 mg, yield 32.59%), in an elution system (DCM:MeOH = 100:1 to 30:1).1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),10.57(s,1H),8.74(s,1H),8.53(s,1H),8.3 1(s,1H),8.22(s,3H),8.08(d,J=12.7Hz,2H),7.93(s,1H),7.77(s,1H),7.53(s,1H ),7.35(s,1H),7.15(s,1H),6.97(s,1H),6.73(s,1H),6.56(d,J=14.4Hz,1H),6.10 (s,1H),4.85(s,2H),3.62(s,2H),3.51(s,4H),2.59(s,3H),2.42(d,J=30.3Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ166.4,165.2,163.8,155.4,151.5,151.4,144.2,140.1, 138.8,135.1,134.3,133.5,132.7,132.6,132.0,131.8,131.0,130.4,129.2,128 .6,127.5,125.7,124.0,123.9,122.7,121.1,117.4,114.5,114.0,112.3,109.3, 105.3,92.4,88.8,66.9,60.2,57.8,53.3,52.9,44.8,20.9.HRMS(ESI)calcd.for C 41 H 34 F3N7O4746.2697[M+H] + ,found746.2697.

[0132] Example 7

[0133]

[0134] Following a similar synthetic method as step eight of Example 1, compound Bal-12 was obtained by reacting 4-trifluoromethylcinnamic acid with compound 10 as a white solid (34 mg, yield 37.65%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1HNMR(400MHz,DMSO-d6)δ13.95(s,1H),10.57(s,1H),8.74(s,1H),8.53(s,1H),8.23(s,3H),8.11(s,1H) ,7.94(s,3H),7.75(s,3H),7.53(s,2H),7.44(s,1H),3.75(s,2H),3.63(s,4H),2.59(s,3H),2.45(s,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,164.4,151.5,151.0,144.2,140.2,139.7,13 8.8,134.3,133.5,132.6,132.1,131.9,131.0,130.5,130.1,129.8,129.6,1 29.3,129.1,128.6,126.0,125.5,124.0,123.7,122.7,121.7,117.7,114.5, 112.3,92.4,88.8,57.7,53.8,53.0,45.6,42.3,20.9.HRMS(ESI)calcd.forC 38 H 30 F6N6O2717.2407[M+H] + ,found 717.2407.

[0135] Example 8

[0136]

[0137] Following a similar synthetic method as step eight of Example 1, compound Bal-22 was obtained by reacting 2-fluoroacrylic acid and compound 10 as a yellow solid (23 mg, yield 28.63%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.59(s,1H),8.74(s,1H),8.53(s,1H),8.22(d,J=9.7Hz,2H),8.12(s,1H),7.94(d,J=7.9Hz,1H),7.76(s ,1H),7.54(d,J=8.0Hz,1H),7.20(s,1H),6.89(s,1H),6.68(d,J=23.9H z,1H),5.32(s,2H),3.63(s,2H),3.54(s,4H),2.59(s,3H),2.44(s,4H). 13C NMR (151MHz, DMSO-d6) δ165.2,160.7,160.5,157.3,155.6,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.0,131.9,131.0,130. 4,128.6,128.1,125.7,124.0,122.7,117.8,117.8,114.5,112.3,92.4,88.8,72.7,60.7,57.6,53.4,46.8,20.9.HRMS(ESI)calcd.for C 31 H 26 F4N6O2591.2126[M+H] + ,found 591.2125.

[0138] Example 9

[0139]

[0140] Compound 10 (20 mg, 0.038 mmol) was weighed and dissolved in tetrahydrofuran (3 mL). Triethylamine (8 μL, 0.057 mmol) was added, followed by dropwise addition of acryloyl chloride (3 μL, 0.038 mmol) under an ice bath at 0 °C. After the addition was complete, the mixture was allowed to react at room temperature for 2 hours. After the reaction was complete, the solvent was evaporated under vacuum, and the mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 25:1) to give a white solid HTY-4015 (15 mg, yield 68.98%). 1 H NMR(400MHz,DMSO-d6)δ13.93(s,1H),10.56(s,1H),8.74(s,1H),8.53(s,1H),8. 23(s,2H),8.20(s,1H),8.09(d,J=8.7Hz,1H),7.93(d,J=8.0Hz,1H),7.75(d,J=8. 6Hz,1H),7.53(d,J=8.1Hz,1H),6.79(dd,J=16.8,10.6Hz,1H),6.10(d,J=16.6Hz ,1H),5.67(d,J=10.5Hz,1H),3.61(s,2H),3.56(s,4H),2.59(s,3H),2.39(s,4H). 13C NMR (151MHz, DMSO-d6) δ165.2,164.7,151.5,151.1,144.2,139.3,138.8,133.5,132.6,132.1,131.9,131.0,130.5,130.1,128. 7,128.1,127.9,127.8,125.7,124.0,122.7,114.5,112.3,92.4,88.8,57.7,53.6,52.9,45.6,42.0,20.9.HRMS(ESI)calcd.for C 31 H 27 F3N6O2573.2220[M+H] + ,found 573.2220.

[0141] Example 10

[0142]

[0143] In the first step, methylamine hydrochloride (540 mg, 8 mmol) and triethylamine (2.2 mL, 16 mmol) were dissolved in THF (6 mL) at room temperature and stirred for 30 minutes under argon protection. A solution of methyl 4-(chlorosulfonyl)benzoate (939 mg, 4 mmol) in THF (4 mL) was added dropwise at 0 °C. The mixture was stirred overnight at room temperature. After dilution with water, the aqueous phase was extracted with ethyl acetate. The combined organic layers were washed with saturated brine and dried over Na₂SO₄. After removing the solvent under vacuum, the mixture was purified by silica gel column chromatography (eluting with PE solution of 10–40% EA) to give compound 10⁻² (450 mg, 49% yield). 1 H NMR (400MHz, CDCl3) δ8.23-8.14(m,2H),7.98-7.89(m,2H),4.57(q,J=5.1Hz,1H),3.96(s,3H),2.69(d,J=5.3Hz,3H).

[0144] In the second step, compound 10⁻² (824.4 mg, 3.60 mmol), K₃PO₄ (1.5 g, 7.21 mmol), CuSO₄ (114 mg, 0.72 mmol), and 1,10-o-phenanthroline (324 mg, 1.8 mmol) were weighed and dissolved in 10 mL of anhydrous toluene. The reaction mixture was degassed using a nitrogen stream, followed by the addition of (2-bromoethynyl)triisopropylsilane (1.14 mL, 4.68 mmol), and the mixture was heated in an oil bath at 70 °C for 12 hours. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with ethyl acetate and washed with H₂O. The aqueous layer was then extracted twice with ethyl acetate, and the collected ethyl acetate layers were washed with saturated brine. Finally, the mixture was dried over anhydrous Na₂SO₄, concentrated under vacuum, and purified by silica gel column chromatography (eluting with 1–5% methanol in DCM solution) to give compound 10⁻³ (743 mg, 90% yield). 1 HNMR (400MHz, CDCl3) δ8.25-8.14(m,2H),8.04-7.92(m,2H),3.98(s,3H),3.11(s,3H),1.04(s,21H).

[0145] In the third step, lithium hydroxide monohydrate (84 mg, 2 mmol) was added to a THF / H₂O (5 mL, 1:1) solution of compound 10⁻³ (410 mg, 1 mmol), and the mixture was stirred for 2 hours. The reaction mixture was acidified to below pH 2 using 1 M HCl and extracted with ethyl acetate. The combined organic layers were dried under vacuum to give compound 10⁻⁴ (387 mg, 98% yield). 1 HNMR (400MHz, DMSO-d6) δ8.26-8.11(m,2H),8.07-7.92(m,2H),3.12(s,3H),0.98(s,21H).

[0146] In the fourth step, following a similar synthetic method as step eight in Example 1, compound 10-4 reacted with compound 10. After the reaction, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was evaporated under vacuum to obtain compound 10-5, which was used directly in the next step without purification. Compound 10-5 (31 mg, 0.034 mmol) was dissolved in tetrahydrofuran (3 mL), and 1 M tetrabutylammonium fluoride solution (0.069 mL, 0.068 mmol) was added dropwise in an ice bath at 0 °C. After the addition was complete, the mixture was allowed to react at room temperature for 4 hours. After the reaction, the solvent was evaporated under vacuum, and the mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain a yellow solid HTY-4019 (24 mg, yield 95.49%).1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d ,J=2.0Hz,1H),8.24-8.21(m,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1 H),7.93(td,J=7.1,2.0Hz,3H),7.77-7.70(m,3H),7.53(d,J=8.2Hz,1H),3.82(s, 1H),3.68(s,2H),3.64(s,2H),3.32(s,4H),3.07(s,3H),2.59(s,3H),2.40(s,2H). 13 C NMR(151MHz,DMSO-d6)δ167.7,165.2,151.5,151.0,144.2,142.0,138.8,1 36.3,134.3,133.5,132.6,132.0,131.9,131.0,130.5,130.1,128.6,128.6 ,128.3,128.1,127.9,125.7,124.0,123.9,122.7,114.5,112.3,92.4,88.8 ,77.6,60.5,57.7,53.3,52.7,47.5,42.1,22.6,20.9.HRMS(ESI)calcd.for C 38 H 32 F3N7O4S 740.2261[M+H] + ,found740.2261.

[0147] Example 11

[0148]

[0149] In the first step, 4-formylbenzoic acid (1500 mg, 10.0 mmol) and nitromethane (1892 mg, 31.0 mmol) were added in a single batch to a solution of ammonium acetate (1945 mg, 25.0 mmol) in acetic acid (20 mL). The mixture was heated under reflux for 4 hours. After the reaction was complete, the reactants were cooled to room temperature and then poured into ice water to precipitate the solid. The mixture was filtered through diatomaceous earth and washed with water to give compound 11-2 (1460 mg, 75% yield). 1 H NMR (400MHz, DMSO-d6) δ10.40(s,1H),8.05(s,2H),7.71(d,J=8.7Hz,2H),6.85(d,J=8.7Hz,2H).

[0150] In the second step, following a similar synthesis method as step eight in Example 1, compound HTY-4022 was obtained by reacting compound 11-2 and compound 10. The compound was a yellow solid (60 mg, yield 66.57%), with an elution system (DCM:MeOH = 100:1 to 50:1). 1 H NMR (400MHz, DMSO-d6) δ13.93(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=1.9H z,1H),8.27(d,J=13.6Hz,1H),8.22(d,J=1.6Hz,2H),8.19(d,J=2.0Hz,1H),8.16(d,J=13. 7Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.92(d,J=8.4Hz,3H),7.75(d,J=8.6Hz,1H),7.53(d ,J=8.2Hz,1H),7.49(d,J=8.3Hz,2H),3.63(s,4H),3.32(s,4H),2.59(s,3H),2.40(s,2H). 13 C NMR (151MHz, DMSO-d6) δ168.5,165.2,151.5,151.0,144.2,139.5,139.3,138.8,134.3,133.5,132.6,132.0,131.8,131.8,131.0,130.5,130. 4,128.6,128.1,127.9,125.7,124.0,123.9,122.7,117.8,114.5,112.3,92.4,88.8,57.7,49.1,31.8,30.8,22.6,20.9.HRMS(ESI)calcd.for C 37 H 30 F3N7O4694.2384[M+H] + ,found694.2385.

[0151] Example 12

[0152]

[0153] Following a similar synthetic method as step eight in Example 1, compound HTY-4023 was obtained by reacting 4-(fluorosulfonyl)benzoic acid with compound 10 as a yellow solid (32 mg, yield 34.95%), using an elution system (DCM:MeOH = 100:1 to 60:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.55 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.5 3(d,J=2.1Hz,1H),8.21(dq,J=9.9,1.6Hz,5H),8.09(dd,J=8.5,2.2Hz,1H),7.9 2(dd,J=8.0,2.0Hz,1H),7.79(d,J=8.3Hz,2H),7.75(d,J=8.6Hz,1H),7.53(d, J=8.2Hz,1H),3.68(s,2H),3.64(s,2H),3.30(s,4H),2.59(s,3H),2.39(s,2H). 13 C NMR (151MHz, DMSO-d6) δ167.2,165.2,151.5,151.0,144.2,144.1,138.8,134.3,133.5,132.6,132.6,132.4,132.0,131.9,131.0,130.5,130. 1,129.3,129.1,128.6,128.1,127.9,125.7,124.0,122.7,114.5,112.3,92.4,88.8,57.7,53.2,52.7,47.5,42.1,20.9.HRMS(ESI)calcd.forC 35 H 28 F4N6O4S 705.19O2[M+H] + ,found705.1902.

[0154] Example 13

[0155]

[0156] Following a similar synthetic method as step eight of Example 1, compound HTY-4024 was obtained by reacting 2-butynedic acid with compound 10 as a yellow solid (60 mg, yield 79.23%), using an elution system (DCM:MeOH = 100:1 to 60:1). 1H NMR (400MHz, CDCl3) δ8.69-8.63(m,2H),8.23(d,J=1.9Hz,1H),8.13(s,1H),8.04(d ,J=2.0Hz,1H),7.99(d,J=9.6Hz,1H),7.94(d,J=2.3Hz,1H),7.81(dd,J=7.9,2.0Hz ,1H),7.73(d,J=8.5Hz,1H),7.34(d,J=8.1Hz,1H),3.74(t,J=5.0Hz,2H),3.62(d,J =7.4Hz, 4H), 2.56 (s, 3H), 2.49 (t, J = 5.1Hz, 2H), 2.41 (t, J = 5.2Hz, 2H), 1.98 (s, 3H). 13 C NMR (151MHz, CDCl3) δ160.5,148.5,146.9,145.7,139.7,134.6,132.5,129.5,128.2,128.0,127.3,126.7,125.7,125.4,124.7,1 22.8,118.7,118.4,113.1,109.9,109.3,108.7,86.6,85.1,84.1,53.0,48.5,47.6,42.2,36.7,24.9,16.1.HRMS(ESI)calcd.forC 32 H 27 F3N6O2585.2220[M+H] + ,found 585.2220.

[0157] Example 14

[0158]

[0159] Methyl 2,3-epoxypropionate (102 mg, 1 mmol) was dissolved in methanol (3 mL), and potassium hydroxide (67 mg, 1.2 mmol) was added. The mixture was stirred overnight at room temperature. After the reaction was complete, the solvent was evaporated under vacuum, and the product was washed with anhydrous diethyl ether to obtain a white solid compound 14-2. This compound was used directly in the next step without purification. Following a similar synthetic method to step eight of Example 1, compound 14-2 reacted with compound 10 to give compound HTY-4025, a pale green solid (32 mg, yield 36.26%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.1 Hz,1H),8.23(s,2H),8.20(s,1H),8.10(d,J=7.9Hz,1H),7.93(d,J=7.9Hz,1H),7.76(d,J =8.6Hz,1H),7.53(d,J=8.1Hz,1H),3.85(dd,J=4.4,2.6Hz,1H),3.62(s,5H),3.49(s,2H) ,2.89(dd,J=6.3,4.3Hz,1H),2.76(dd,J=6.3,2.6Hz,1H),2.59(s,3H),2.46-2.35(m,4H). 13 C NMR(151MHz,DMSO-d6)δ166.0,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.9,131.0,130.5,128.6,128.1, 127.9,125.7,124.0,123.9,122.7,114.5,112.3,92.4,88.8,57.7,53.4,52.7,47.4,46.0,44.9,42.0,20.9.HRMS(ESI)calcd.for C 31 H 27 F3N6O3589.2169[M+H] + ,found589.2169.

[0160] Example 15

[0161]

[0162] Following the synthetic route described above and referring to the synthetic method of Example 10, compound HTY-4030 was obtained as a white solid (14 mg, yield 19.90%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.55 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.53 (d,J=2.0Hz,1H),8.22(d,J=1.6Hz,2H),8.19(d,J=2.0Hz,1H),8.09(d,J=8.5Hz ,1H),8.01(d,J=8.3Hz,2H),7.92(dd,J=8.0,1.9Hz,1H),7.75(d,J=8.5Hz,1H), 7.53(dd,J=8.4,3.2Hz,3H),3.87(s,3H),3.63(s,3H),2.59(s,3H),2.39(s,2H). 13 C NMR(151MHz,DMSO-d6)δ168.4,166.2,165.2,151.5,151.0,144.2,140.9,138 .8,134.3,133.5,132.6,132.2,132.1,132.0,131.8,131.0,130.8,130.5,13 0.1,129.8,129.1,128.6,127.7,125.7,124.0,122.7,114.5,112.3,92.4,88 .8,67.9,57.7,53.3,52.8,47.6,42.1,38.5,22.6,20.9.HRMS(ESI)calcd.for C 39 H 32 F3N7O3704.2591[M+H] + ,found 704.2307.

[0163] Example 16

[0164]

[0165] In the first step, potassium carbonate (4.00 g, 28.90 mmol) was added to a round-bottom flask containing an EtOH / H₂O (1:1, 50 mL) solution of hydroxylamine hydrochloride (4.00 g, 57.80 mmol). The mixture was stirred at room temperature for 5 minutes. Ethyl 3-oxophenylpropionate (10.0 mL, 57.80 mmol) was added sequentially, and the reaction mixture was heated to 65 °C and reacted for 10 hours. After the reaction was completed, the mixture was quenched with water, extracted three times with DCM, and the combined organic layers were washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under vacuum. The product was purified by silica gel column chromatography (PE:EA = 20:1 to 5:1) to give compound 16-2 (5.10 g, 55% yield). 1HNMR (400MHz, CDCl3) δ7.71-7.66(m,2H),7.56-7.46(m,3H),3.81(s,2H).

[0166] In the second step, triethylamine (1430 μL, 10.30 mmol) was added dropwise to a suspension of compound 16-2 (2.00 g, 12.40 mmol) in POCl3 (16.5 mL, 176.80 mmol) at 0 °C. The mixture was stirred at 105 °C for 12 hours, poured into a saturated ice solution of NaHCO3, and extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous Na2SO4, and concentrated under vacuum. Purification by silica gel column chromatography (PE:EA = 10:1) yielded compound 16-3 (1.80 g, 81% yield). 1 H NMR (400MHz, CDCl3) δ7.76 (dd, J=6.8, 3.0Hz, 2H), 7.47 (dd, J=4.8, 1.9Hz, 3H), 6.48 (s, 1H).

[0167] In the third step, under argon atmosphere, anhydrous FeCl2 (203 mg, 1.60 mmol) was added to a dry acetonitrile (40 mL) solution of compound 16-3 (718 mg, 4.00 mmol). The mixture was stirred at room temperature for 8 hours. Water (50 mL) was added and stirred at room temperature for 15 minutes. The mixture was extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous Na2SO4, and concentrated under vacuum. Washing with a mixture of n-hexane and diethyl ether (10:1) gave compound 16-4 (612 mg, 95% yield). 1 HNMR (400MHz, CDCl3) δ7.92-7.88(m,2H),7.68-7.64(m,1H),7.61-7.56(m,2H),2.85(s,1H).

[0168] In the fourth step, weigh compound 16-4 (26 mg, 0.162 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (38 mg, 0.202 mmol), and 1-hydroxybenzotriazole (36 mg, 0.27 mmol) into a reaction flask. Add N,N-diisopropylethylamine (0.067 mL, 0.405 mmol) and anhydrous N,N-dimethylformamide (3 mL), and stir for 5 minutes. Dissolve compound 10 (70 mg, 0.135 mmol) in 1 mL of anhydrous N,N-dimethylformamide, add N,N-diisopropylethylamine (0.033 mL, 0.202 mmol) for alkalization, and then add it to the reaction solution. React overnight at room temperature. After the reaction was completed, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 60:1) to obtain a yellow solid compound HTY-4031 (27 mg, yield 30.33%). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(d,J=7.8Hz,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz ,1H),8.25-8.20(m,3H),8.12(dd,J=8.5,2.3Hz,1H),7.94(dd,J=8.0,2.0Hz,1H),7.89-7.87(m,1H), 7.79(d,J=8.6Hz,1H),7.77-7.71(m,1H),7.66(m,2H),7.53(dd,J=8.4,3.1Hz,1H),7.45-7.38(m,1H) ,3.92(s,2H),3.66(s,2H),3.63(s,1H),3.52(s,2H),2.59(d,J=2.3Hz,3H),2.56(s,2H),2.41(s,2H). 13 C NMR(151MHz,DMSO-d6)δ168.7,165.2,159.2,151.5,151.0,144.2,138.8,134.3,134.2,133.5,132.7,131.9,131.0,130.5,1 30.4,130.0,128.9,128.7,127.4,114.5,112.3,92.4,88.8,57.8,53.7,52.9,45.4,42.4,28.3,20.9.HRMS(ESI)calcd.forC 37 H 30F3N7O2662.2486[M+H] + ,found 662.2487.

[0169] Example 17

[0170]

[0171] Following a similar synthetic method as step eight of Example 1, compound HTY-4069 was obtained by reacting 4,4,4-trifluorobutenoic acid with compound 10 as a yellow solid (15 mg, yield 18.02%), using an elution system (DCM:MeOH = 100:1 to 50:1). 1 H NMR (400MHz, DMSO) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J= 2.0Hz,1H),8.23(t,J=2.2Hz,2H),8.20(d,J=2.0Hz,1H),8.10(d,J=8.5Hz,1H),7.9 3(dd,J=8.0,1.9Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),7.36(dd,J= 15.5,2.4Hz,1H),6.76(m,1H),3.62(s,2H),3.58(s,4H),2.59(s,3H),2.42(s,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,162.1,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.0,131.9,131.0,131.0,130.9,130.5,1 30.1,128.6,127.5,127.3,124.0,122.7,117.7,114.5,112.3,92.4,88.8,57.6,53.4,52.7,45.9,42.3,20.9.HRMS(ESI)calcd.forC 32 H 26 F6N6O2641.2094[M+H] + ,found641.2095.

[0172] Example 18

[0173]

[0174] Following a similar synthetic method as in Example 9, compound HTY-4092 was obtained by reacting dichloroacetyl chloride with compound 10 as a yellow solid (27 mg, yield 28.65%), using an elution system (DCM:MeOH = 100:1 to 50:1). 1 H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.5 3(d,J=2.0Hz,1H),8.23(t,J=2.2Hz,2H),8.20(d,J=1.9Hz,1H),8.10(dd,J=8.6 ,2.2Hz,1H),7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1 Hz,1H),7.24(s,1H),3.62(s,2H),3.59-3.50(m,4H),2.59(s,3H),2.43(m,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,162.1,151.5,151.0,144.2,138.8,134.3,133.5,132.6,131.9,131.0,130.5,130.1,128.7, 125.7,124.6,124.0,122.7,117.8,114.5,112.3,92.4,88.8,66.3,57.6,52.9,52.6,46.2,43.2,20.9.HRMS(ESI)calcd.for C 30 H 25 Cl2F3N6O2629.1441[M+H] + ,found 629.1441.

[0175] Example 19

[0176]

[0177] Following a similar synthetic method as step eight in Example 1, compound HTY-4093 was obtained by reacting dibromoacetic acid with compound 10 as a yellow solid (15 mg, yield 18.02%), using an elution system (DCM:MeOH = 100:1 to 70:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 1.9Hz, 1H), 8.53 (d,J=2.0Hz,1H),8.26-8.21(m,2H),8.20(d,J=1.9Hz,1H),8.10(d,J=9.0Hz,1H) ,7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),7.14( s,1H),3.62(s,2H),3.55(d,J=16.1Hz,4H),2.59(s,3H),2.43(d,J=22.3Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,162.6,151.5,151.0,144.2,138.8,134.3,133.5,132.6,131.9,131.9,131.0,130.5,130 .1,128.7,124.6,124.0,122.7,114.5,112.3,92.4,88.8,57.6,52.9,52.6,46.7,43.3,38.8,20.9.HRMS(ESI)calcd.for C 30 H 25 Br2F3N6O2717.0431[M+H] + ,found717.0432.

[0178] Example 20

[0179]

[0180] Following a similar synthetic method as step eight of Example 1, compound HTY-4095 was obtained by reacting chlorofluoroacetic acid with compound 10 as a yellow solid (27 mg, yield 29.40%), using an elution system (DCM:MeOH = 100:1 to 50:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.53 ( d,J=2.0Hz,1H),8.23(t,J=2.2Hz,2H),8.20(d,J=1.9Hz,1H),8.12-8.07(m,1H), 7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),7.28(d ,J=48.7Hz,1H),3.63(s,2H),3.55-3.43(m,4H),2.59(s,3H),2.47-2.39(m,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,162.1,161.9,151.5,151.0,144.2,138.8,134.3,133.5,132.6,131.9,131.0,130.5,128.6,124 .4,124.0,122.7,117.8,114.5,112.3,105.8,92.6,92.4,91.0,88.8,57.6,54.1,52.5,45.3,42.3,20.9.HRMS(ESI)calcd.for C 30 H 25 ClF4N6O2613.1736[M+H] + ,found 613.1757.

[0181] Example 21

[0182]

[0183] Following a similar synthetic method as step eight in Example 1, compound HTY-4096 was obtained by reacting bromochloroacetic acid with compound 10 as a yellow solid (33 mg, yield 29.40%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.72( dd,J=4.4,1.5Hz,1H),8.53(d,J=2.0Hz,1H),8.23(s,2H),8.19(d,J=2.0Hz,1H),8 .09(dd,J=8.5,2.2Hz,1H),7.93(dd,J=8.0,1.9Hz,1H),7.75(d,J=8.7Hz,1H),7.5 3(d,J=8.2Hz,1H),3.62(s,2H),3.54(s,4H),2.59(s,3H),2.43(d,J=17.9Hz,5H). 13 C NMR(151MHz,DMSO-d6)δ165.2,162.5,162.1,151.5,151.0,144.2,138.8,134.3,133.5,132.6,131.9,131.0,130.5,130.1, 128.6,125.7,124.0,122.7,117.7,114.5,112.3,92.4,88.8,66.3,57.6,54.1,52.9,52.6,46.3,20.9.HRMS(ESI)calcd.for C 30 H 25 BrClF3N6O2673.0936[M+H] + ,found673.0934.

[0184] Example 22

[0185]

[0186] Following a similar synthetic method as step eight of Example 1, compound HTY-4103 was obtained by reacting difluoroacetic acid with compound 10 as a white solid (20 mg, yield 32.43%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.53 (d, J=1.7Hz,1H),8.26-8.21(m,2H),8.20(d,J=2.0Hz,1H),8.10(dd,J=8.5,2.2Hz,1H), 7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.6Hz,1H),7.53(d,J=8.2Hz,1H),6.73(t,J= 52.7Hz,1H),3.63(s,2H),3.53(d,J=5.1Hz,4H),2.59(s,3H),2.44(p,J=4.7Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,160.4,151.5,151.0,144.2,138.8,134.3,133.5,132.6,131.9,131.0,130.5,130.1,128.6,124.0,1 23.9,122.7,117.8,117.7,114.5,112.3,109.5,107.9,106.3,92.4,88.8,57.6,53.1,52.5,44.9,42.3,20.9.HRMS(ESI)calcd.forC 30 H 25 F5N6O2597.2032[M+H] + ,found597.2049.

[0187] Example 23

[0188]

[0189] In the first step, methyl 3-iodo-4-methylbenzoate (662.5 mg, 2.4 mmol), 3-alkynylimidazole [1,2-B]pyridazine (286.3 mg, 2.0 mmol), tetrakis(triphenylphosphine)palladium (115.5 mg, 0.1 mmol), and cuprous iodide (38.09 mg, 0.2 mmol) were weighed into a reaction flask. Under argon protection, N,N-diisopropylethylamine (0.66 mL, 4.0 mmol) and N,N-dimethylformamide (5 mL) were added, and the reaction was carried out overnight at 120 °C. After the reaction was completed, the mixture was filtered, and the filter cake was washed with acetonitrile to obtain yellow compound 12, which could be used directly in the next step without purification. 1H NMR (400MHz, DMSO-d6) δ8.72(s,1H),8.26(s,2H),8.06(s,1H),7.89(d,J=9.9Hz,1H),7.54(d,J=9.0Hz,1H),7.39(s,1H),3.87(s,3H),2.59(s,3H).

[0190] In the second step, following a similar synthesis method as step six of Example 1, compound 13 was obtained by reacting compound 12 and compound 4 as a yellow solid (80 mg, yield 65.04%), using an elution system (PE:EA = 5:1 to 1:1). 1 H NMR (400MHz, DMSO-d6) δ10.57(s,1H),8.73(dd,J=4.4,1.5Hz,1H),8.28-8.21(m,4H),8.08(dd,J=8.6,2.2Hz,1H),7.95(dd,J=8.0,2.0Hz,1H),7. 74(d,J=8.5Hz,1H),7.55(d,J=8.1Hz,1H),7.40(dd,J=9.1,4.3Hz,1H),3 .59(s,2H),3.32(s,5H),2.61(s,3H),2.34(t,J=5.0Hz,4H),1.39(s,9H).

[0191] In the third step, compound 13 (500 mg, 0.8 mmol) was weighed into a reaction flask, and 5 mL of 1,4-dioxane solution in 4 M hydrochloric acid was added. The reaction was carried out at room temperature for 4 hours. After the reaction was completed, the solvent was evaporated under vacuum. No purification was required, and the solvent was used directly in the next step to obtain yellow compound 14.

[0192] Fourth step, following a similar synthesis method as in Example 9, compound HTY-4045 was obtained by reacting acryloyl chloride and compound 14 as a yellow solid (22 mg, yield 20.23%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.73(dd,J=4.5,1.5Hz,1H),8.24(m,4H),8.09(dd ,J=8.5,2.2Hz,1H),7.95(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.6Hz,1H),7.55(d,J=8.1Hz ,1H),7.39(dd,J=9.2,4.4Hz,1H),6.79(dd,J=16.7,10.5Hz,1H),6.10(dd,J=16.7,2.5H z,1H),5.67(dd,J=10.5,2.4Hz,1H),3.61(s,2H),3.56(s,4H),2.61(s,3H),2.39(s,4H). 13 C NMR(151MHz,DMSO-d6)δ165.1,164.7,145.5,144.0,140.1,138.8,138.7,132.7,132.1,131.9,130.6,130.6,130.1,129.0,128.6,1 28.1,127.8,126.6,125.7,124.0,123.9,122.2,119.6,112.2,96.9,81.6,57.7,53.6,52.9,45.6,42.0,20.9.HRMS(ESI)calcd.for C 31 H 27 F3N6O2573.2220[M+H] + ,found573.2220.

[0193] Example 24

[0194]

[0195] Following a similar synthetic method as step eight of Example 1, compound HTY-4046 was obtained by reacting 2-cyano-3-cyclopropylacrylic acid and compound 14 as a yellow solid (263 mg, yield 27.20%), using an elution system (DCM:MeOH = 100:1 to 80:1). 1H NMR (400MHz, DMSO-d6) δ10.58 (s, 1H), 8.73 (dd, J = 4.4, 1.6Hz, 1H), 8.29-8.20 (m, 4 H),8.10(d,J=8.5Hz,1H),7.95(dd,J=8.0,1.9Hz,1H),7.75(d,J=8.6Hz,1H),7.55 (d,J=8.1Hz,1H),7.40(dd,J=9.2,4.5Hz,1H),6.59(d,J=11.1Hz,1H),3.62(s,2H) ,3.53(s,4H),2.61(s,3H),2.43(s,4H),1.17(d,J=4.6Hz,1H),1.00-0.73(m,4H). 13 C NMR(151MHz,DMSO-d6)δ165.1,164.2,162.0,145.6,144.0,140.1,138.8,138.7,132.7,132.0,131.8,130.6,130.6,130.1,129.0,126.6,12 4.0,123.9,122.2,119.6,116.2,112.2,106.6,105.9,96.9,81.6,66.8,64.2,57.7,52.8,49.1,20.9,15.8,14.4,10.4.HRMS(ESI)calcd.for C 35 H 30 F3N7O2638.2486[M+H] + ,found 638.2485.

[0196] Example 25

[0197]

[0198] Following a similar synthetic method as step eight of Example 1, compound HTY-4048 was obtained by reacting (E)-4-(dimethylamino)but-2-enoic acid with compound 14 as a yellow solid (20 mg, yield 23.54%), using an elution system (DCM:MeOH = 100:1 to 15:1). 1H NMR (400MHz, DMSO-d6) δ10.58 (s, 1H), 8.73 (dd, J = 4.4, 1.6Hz, 1H), 8.29-8.20 (m, 4H),8.09(dd,J=8.5,2.2Hz,1H),7.95(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.6Hz,1 H),7.55(d,J=8.2Hz,1H),7.40(dd,J=9.2,4.4Hz,1H),6.59(d,J=2.4Hz,2H),3.6 1(s,2H),3.55(s,4H),3.06-3.01(m,2H),2.61(s,3H),2.39(s,4H),2.15(s,6H). 13 C NMR(151MHz,DMSO-d6)δ165.1,164.5,145.6,144.0,142.2,140.1,138.8,138.7,132.7,132.1,131.9,130.6,130.6,129.0,128.0,127.8 ,126.6,124.0,122.8,122.2,119.6,117.8,112.2,96.9,81.6,60.3,57.7,53.6,53.0,45.4,42.0,31.8,30.8,20.9.HRMS(ESI)calcd.for C 34 H 34 F3N7O2630.2799[M+H] + ,found 630.2792.

[0199] Example 26

[0200]

[0201] Following a similar synthetic method as step eight of Example 1, compound HTY-4049 was obtained by reacting 2-butynedic acid with compound 14 as a yellow solid (35 mg, yield 39.93%), using an elution system (DCM:MeOH = 100:1 to 80:1). 1H NMR(400MHz,DMSO-d6)δ10.58(s,1H),8.73(dd,J=4.4,1.6Hz,1H),8.28-8.21(m,4H), 8.09(dd,J=8.5,2.2Hz,1H),7.95(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.55( d,J=8.2Hz,1H),7.40(dd,J=9.2,4.4Hz,1H),3.71-3.66(m,2H),3.62(s,2H),3.49(t, J=5.1Hz,2H),2.61(s,3H),2.43(t,J=5.0Hz,2H),2.35(t,J=5.2Hz,2H),2.01(s,3H). 13 CNMR(151MHz,DMSO-d6)δ165.1,152.4,145.6,144.0,140.1,138.8,138.7,132.7,132.0,131.9,130.6,130.6,129.0,128.1,127.9 ,126.6,125.7,124.0,122.2,119.6,112.2,96.9,90.0,81.6,73.3,57.6,53.4,52.6,46.8,41.3,20.9,3.8.HRMS(ESI)calcd.forC 32 H 27 F3N6O2585.2220[M+H] + ,found585.2220.

[0202] Example 27

[0203]

[0204] In the first step, a mixture of compound 27-1 (1 g, 2.0 mmol), tert-butyl pyrrolidone-3-ylcarbamate (R) (440 mg, 2.4 mmol), and DIPEA (0.72 mL, 4.0 mmol) in isopropanol (15 mL) was refluxed at 140 °C for 2 hours. Then, it was extracted with ethyl acetate (100 mL) and washed with saturated brine. The solvent was evaporated under vacuum, and the mixture was used directly in the next step without purification to give compound 27-2 (851 mg, 76% yield).

[0205] In the second step, the mixture of compound 27-2 (840 mg, 1.5 mmol) and TFA (3 mL) in DCM (3 mL) was stirred overnight at room temperature. The mixture was then neutralized to pH 12 with 1 N NaOH solution, extracted twice with DCM, dried over anhydrous sodium sulfate, and finally the solvent was evaporated under vacuum. No purification was required, and the mixture was used directly in the next step to give compound 27-3 (565 mg, 82% yield).

[0206] In the third step, compound 27-3 (800 mg, 1.7 mmol), 1-(2-tetrahydropyranyl)-1H-pyrazole-5-boronate pinacol ester (600 mg, 2 mmol), Pd(PPh3)4 (100 mg, 0.09 mmol), K3PO4 (720 mg, 3.4 mmol), and toluene (20 mL) were added to a 50 mL round-bottom flask. The mixture was stirred overnight at 110 °C under argon protection. After extraction with ethyl acetate and washing with saturated brine, the mixture was purified by silica gel rapid column chromatography (DCM / MeOH = 20:1) to give compound 27-4 (624 mg, 69% yield).

[0207] In the fourth step, compound 27-4 (400 mg, 0.75 mmol) was dissolved in a mixed solution of methanol (2 mL) and tetrahydrofuran (2 mL), followed by the addition of concentrated hydrochloric acid (1.5 mL), and the reaction was carried out overnight at room temperature. After the reaction was completed, saturated sodium bicarbonate solution was added to the reaction solution for quenching, the aqueous phase was extracted three times with ethyl acetate, the organic phases were combined, anhydrous sodium sulfate was added for drying, and the solvent was evaporated under vacuum to obtain compound 27-5, which did not require purification and was used directly in the next step. Following a similar synthetic method to step eight of Example 1, compound HTY-4063 was obtained by reacting 2-cyano-3-cyclopropylacrylic acid and compound 27-5 as a yellow solid (22 mg, yield 35.27%), using an elution system (DCM:MeOH = 100:1 to 40:1). 11H NMR (400 MHz, DMSO-d6) δ 12.93 (s, 1H), 10.21 (s, 1H), 8.77 - 8.73 (m, 1H), 8.27 (d, J = 6.5 Hz, 1H), 8.06 (d, J = 2.4 Hz, 1H), 7.87 (d, J = 9.1 Hz, 2H), 7.33 (d, J = 8.6 Hz, 2H), 6.88 (d, J = 11.1 Hz, 1H), 6.40 (d, J = 2.1 Hz, 1H), 4.24 (q, J = 6.2 Hz, 1H), 3.47 - 3.41 (m, 2H), 3.13 (dd, J = 11.5, 5.4 Hz, 1H), 2.02 (dd, J = 13.1, 6.4 Hz, 2H), 1.86 (m, 2H), 1.35 (s, 1H), 1.30 - 1.25 (m, 2H), 0.98 - 0.88 (m, 2H). 13 13C NMR (151 MHz, DMSO) δ 164.2, 161.4, 145.3, 139.1, 130.1, 127.4, 125.5, 122.3, 121.9, 118.3, 116.1, 108.2, 31.8, 30.8, 30.5, 27.0, 22.6, 15.9, 14.4, 10.8. HRMS (ESI) calcd. for C 27 1 24 17ClF2N7O3 568.1675 [M + H] + , found 568.1670.

[0208] Example 28

[0209]

[0210] In the first step, following a similar synthetic method to Example 9, compound 28-1 was obtained by reacting acrylamide with compound 27-4 as a white solid (54 mg, yield 46.06%), using an elution system (DCM:MeOH = 100:1 to 50:1). ¹H NMR (400 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.84 (t, J = 2.3 Hz, 1H), 8.23 ​​(d, J = 6.4 Hz, 1H), 8.03 (s, 1H), 7.86 (d, J = 9.1 Hz, 2H), 7.61 (d, J = 1.7 Hz, 1H), 7.33 (d, J = 8.7 Hz, 2H), 6.43 (d, J = 2.0 Hz, 1H), 6.18 (m, 1H), 6.07 (m, 1H). ),5.58(dd,J=9.9,2.5Hz,1H),5.08-4.99(m,1H),4.21(s,1H),3.83(t,J=13.2Hz,1H),3.48-3.37(m,2H) ,3.32-3.12(m,4H),2.29(d,J=12.7Hz,1H),2.03-1.90(m,2H),1.78(d,J=13.6Hz,2H),1.58-1.42(m,3H).

[0211] In the second step, compound 28-1 (50 mg, 0.085 mmol) was dissolved in a mixed solution of methanol (2 mL) and tetrahydrofuran (2 mL), followed by the addition of concentrated hydrochloric acid (0.05 mL), and the reaction was allowed to proceed overnight at room temperature. After the reaction was completed, saturated sodium bicarbonate solution was added to the reaction solution for quenching. The aqueous phase was extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The solution was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 15:1) to obtain a white solid compound HTY-4078 (33 mg, yield 77.31%). 1H NMR (400MHz, DMSO-d6) δ12.93(s,1H),10.21(s,1H),8.76(s,1H),8.28(d,J=6.6Hz,1H),8.06(d,J=2.4 Hz,1H),7.87(d,J=9.1Hz,2H),7.33(d,J=8.7Hz,2H),6.41(d,J=2.1Hz,1H),6.19(dd,J=17.0,10.0Hz, 1H), 6.07 (dd, J=17.1, 2.4Hz, 1H), 5.57 (dd, J=10.0, 2.4Hz, 1H), 4.24 (q, J=5.8Hz, 1H), 3.42 (dd, J=11. 5,6.2Hz,4H),3.04(dd,J=11.6,4.6Hz,1H),2.02(dt,J=13.0,6.7Hz,1H),1.77(dt,J=12.7,6.5Hz,1H). 13 C NMR(151MHz,DMSO-d6)δ164.9,145.2,139.4,139.1,131.9,130.1,127.4,125.9, 125.5,123.6,122.3,121.9,118.3,54.7,49.0,47.6,30.8.HRMS(ESI)calcd.for C 23 H 21 ClF2N6O35O3.1410[M+H] + ,found 503.1404.

[0212] Example 29

[0213]

[0214] The first step, following a similar synthetic method to step eight of Example 1, involved reacting 2-butynedic acid and compound 27-4 to obtain compound 29-1, a white solid (90 mg, yield 75.23%), in an elution system (DCM:MeOH = 100:1 to 80:1). 1HNMR (400MHz, DMSO-d6) δ10.21 (s, 1H), 8.83 (s, 1H), 8.68 (d, J = 6.5Hz, 1H), 8.03 (s, 1H), 7. 86(d,J=9.1Hz,2H),7.61(d,J=1.7Hz,1H),7.33(d,J=8.7Hz,2H),6.41(s,1H),5.06(d,J=9 .8Hz,1H),4.14(d,J=6.1Hz,1H),3.85(s,1H),3.42(d,J=9.9Hz,1H),3.27-3.10(m,3H),2. 31(d,J=13.9Hz,1H),1.97(d,J=16.8Hz,2H),1.93(s,3H),1.84-1.71(m,2H),1.45(s,4H).

[0215] In the second step, following a similar synthesis method as in Example 28, compound HTY-4082 was obtained as a white solid (62 mg, yield 86.13%), using an elution system (DCM:MeOH = 100:1 to 20:1). 1 H NMR(400MHz,DMSO-d6)δ12.93(s,1H),10.21(s,1H),8.79-8.67(m,2H),8.05 (d,J=2.4Hz,1H),7.87(d,J=9.1Hz,2H),7.33(d,J=8.6Hz,2H),6.40(d,J=2. 1Hz,1H),4.16(d,J=7.4Hz,1H),3.38(s,2H),3.28(s,1H),3.02(dd,J=11.5, 4.5Hz,1H),1.98(d,J=4.6Hz,1H),1.93(s,3H),1.77(dt,J=12.2,6.3Hz,1H). 13 C NMR (151MHz, DMSO-d6) δ152.8,145.2,139.3,139.1,130.1,127.4,125.5,123.6,122. 3,121.9,118.3,106.5,83.1,76.0,54.4,49.2,47.6,30.3,3.5.HRMS(ESI)calcd.for C 24 H 21 ClF2N6O3515.1410[M+H] + ,found 515.1404.

[0216] Example 30

[0217]

[0218] The first step, following a similar synthetic method to step eight of Example 1, involved reacting BOC-β-alanine with compound 10 to obtain compound 30-2, which was a yellow solid (250 mg, yield 47.72%), in an elution system (DCM:MeOH = 100:1 to 40:1). 1 HNMR(400MHz,DMSO-d6)δ13.93(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d, J=2.0Hz,1H),8.23(s,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.93(d d,J=7.9,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.2Hz,1H),6.69(t,J=5.9Hz, 1H), 3.60 (s, 2H), 3.44 (d, J = 5.5Hz, 4H), 2.59 (s, 3H), 2.46-2.31 (m, 8H), 1.37 (s, 9H).

[0219] In the second step, compound 30-2 (500 mg, 0.8 mmol) was weighed into a reaction flask, and 5 mL of 1,4-dioxane solution in 4 M hydrochloric acid was added. The reaction was carried out at room temperature for 4 hours. After the reaction was completed, the solvent was evaporated under vacuum, and the compound was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to obtain the brown solid compound HTY-4052 (192 mg, yield 90.08%). 1 HNMR (400MHz, DMSO-d6) δ10.57(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H),8.23 (d,J=2.7Hz,2H),8.20(d,J=1.9Hz,1H),8.10(d,J=6.8Hz,1H),7.93(dd,J=8.0,2.0Hz,1H ),7.75(d,J=8.6Hz,1H),7.53(d,J=8.1Hz,1H),3.60(s,2H),3.46(s,4H),3.03(qd,J=7.2 ,5.4Hz,2H),2.78(t,J=6.4Hz,2H),2.59(s,3H),2.44(t,J=6.4Hz,2H),2.40-2.33(m,4H). 13CNMR(151MHz,DMSO-d6)δ169.2,165.2,151.5,151.1,144.2,138.8,134.2,133.5,132.6,131.8,131.0,130.4,128.7 ,124.0,122.7,114.5,112.3,92.4,88.8,57.8,53.4,53.0,45.3,41.5,33.7,23.1,20.9,15.2.HRMS(ESI)calcd.for C 31 H 30 F3N7O2590.2486[M+H] + ,found 590.2475.

[0220] The third step involves a similar synthesis method to that in Example 9, where acryloyl chloride and compound HTY-4052 react to yield compound HTY-4055, which is a white solid (60 mg, yield 55.19%), using an elution system (DCM:MeOH = 100:1 to 25:1). 1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H),8 .23(s,2H),8.19(d,J=2.0Hz,1H),8.15-8.11(m,1H),8.09(d,J=8.6Hz,1H),7.93(dd,J=8.0,2.0Hz,1 H),7.75(d,J=8.6Hz,1H),7.53(d,J=8.1Hz,1H),6.22(dd,J=17.1,10.1Hz,1H),6.06(dd,J=17.1,2.3 Hz,1H),5.56(dd,J=10.1,2.3Hz,1H),3.60(s,2H),3.46(d,J=20.0Hz,8H),2.59(s,3H),2.37(s,4H). 13 C NMR(151MHz,DMSO-d6)δ169.4,165.1,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.2,132.1,131.8,131.0,130.5,130.1,128.6, 125.7,125.5,124.0,123.9,122.7,117.7,114.5,112.3,92.4,88.8,57.8,53.4,52.9,45.3,41.5,35.6,32.8,22.6.HRMS(ESI)calcd.for C34 H 32 F3N7O3644.2591[M+H] + ,found 644.2591.

[0221] Example 31

[0222]

[0223] Following a similar synthetic method as step eight in Example 1, compound HTY-4056 was obtained by reacting 2-cyano-3-cyclopropylacrylic acid and compound HTY-4052. The compound was a grayish-white solid (64 mg, yield 90.36%), with an elution system (DCM:MeOH = 100:1 to 40:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.1Hz,1H),8.53(d,J=2.0Hz,1H),8. 22(s,2H),8.19(d,J=1.9Hz,1H),8.12-8.07(m,2H),7.92(dd,J=7.9,2.0Hz,1H),7.74(d,J=8.5Hz,1 H),7.53(d,J=8.1Hz,1H),6.94(d,J=11.2Hz,1H),3.60(s,2H),3.49-3.42(m,4H),3.32(s,2H),2.59 (s,3H),2.53(d,J=7.2Hz,2H),2.41-2.32(m,4H),1.89(m,1H),1.30-1.23(m,2H),0.97-0.90(m,2H). 13 C NMR(151MHz,DMSO-d6)δ169.4,165.2,164.7,160.7,151.5,151.0,144.2,1 38.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,128.0,127. 9,124.0,123.9,122.7,116.1,114.5,112.3,107.8,92.4,88.8,57.8,56.5 ,53.3,52.9,49.1,36.5,32.5,20.9,19.0,15.9,10.9.HRMS(ESI)calcd.for C 38 H 35 F3N8O3709.2857[M+H] + ,found 709.2855.

[0224] Example 32

[0225]

[0226] Following a similar synthetic method as step eight in Example 1, compound HTY-4060 was obtained by reacting 2-butynedic acid with compound HTY-4052. The compound was a grayish-white solid (66 mg, yield 67.14%), with an elution system (DCM:MeOH = 100:1 to 30:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.1Hz,1H),8.53(d,J=2 .0Hz,1H),8.40(t,J=5.7Hz,1H),8.22(d,J=1.3Hz,2H),8.19(d,J=1.9Hz,1H),8.09(dd, J=8.5,2.3Hz,1H),7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.7Hz,1H),7.53(d,J=8.1Hz ,1H),3.60(s,2H),3.44(d,J=17.2Hz,6H),2.59(s,3H),2.44-2.25(m,6H),1.93(s,3H). 13 CNMR(151MHz,DMSO-d6)δ169.1,165.2,153.0,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.5,130.3,130.1, 128.6,124.0,122.7,117.7,114.5,112.3,92.4,88.8,82.8,76.1,57.8,53.3,52.9,45.3,41.5,35.9,32.3,20.9,3.4.HRMS(ESI)calcd.for C 35 H 32 F3N7O3656.2591[M+H] + ,found 656.2590.

[0227] Example 33

[0228]

[0229] In the first step, 50 mg (0.36 mmol) of 2-cyano-3-cyclopropylacrylic acid and 10 wt% palladium / carbon (10 mg, 0.025 mmol) were weighed into a reaction flask, and 2 mL of anhydrous methanol was added. The flask was then purged with a hydrogen balloon, and the reaction was carried out under hydrogen conditions for 5 hours. After the reaction was completed, the mixture was filtered, the filter cake was washed with EA, and the filtrate was collected. The solvent was evaporated under vacuum to obtain a colorless oily compound 33-2 (49 mg, 99.67%), which could be used directly in the next step without further purification.

[0230] In the second step, following a similar synthesis method as step eight of Example 1, compound 33-2 was reacted with compound 10 to obtain compound HTY-4100, a yellow solid (31 mg, yield 28.53%), in an elution system (DCM:MeOH = 100:1 to 50:1). 1 HNMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0H z,1H),8.23(s,2H),8.20(d,J=1.9Hz,1H),8.10(d,J=8.0Hz,1H),7.93(dd,J=8.0,1.9Hz,1H) ,7.74(t,J=7.6Hz,1H),7.53(d,J=8.1Hz,1H),3.61(d,J=10.4Hz,2H),3.51(s,4H),2.87(s, 1H),2.59(s,3H),2.41(d,J=20.9Hz,5H),2.07-1.92(m,1H),1.76(m,2H),1.62-1.37(m,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,164.3,151.5,151.0,144.2,138.7,134.3,133.5,132.6,131.7,131.0,130.5,130.1,128.6,124.0,1 22.7,119.1,118.5,114.5,112.3,92.4,88.8,57.6,52.9,51.7,47.2,46.0,31.2,29.0,20.9,17.0,14.3,14.2.HRMS(ESI)calcd.for C 35 H 32 F3N7O2640.2642[M+H] + ,found 640.2646.

[0231] Example 34

[0232]

[0233] Compound 10 (77 mg, 0.15 mmol) was weighed into a reaction flask, and 99% formic acid (2 mL) was added. The reaction was carried out at 110 °C for 3 hours. After the reaction was completed, the mixture was quenched with saturated sodium bicarbonate aqueous solution, extracted three times with ethyl acetate, and the organic phases were combined. The mixture was washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 30:1) to give a white solid compound HTY-4111 (55 mg, yield 67.07%). 1 H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 2.1Hz, 1H), 8.53 (d, J=2.0Hz,1H),8.23(d,J=2.5Hz,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz, 1H),8.00(s,1H),7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1 Hz,1H),3.62(s,2H),3.45-3.37(m,4H),2.59(s,3H),2.37(dt,J=23.9,5.1Hz,4H). 13 C NMR (151MHz, DMSO-d6) δ165.2,161.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128. 6,125.7,124.0,122.7,117.8,117.7,114.5,112.3,92.4,88.8,57.8,55.4,53.7,52.6,45.3,20.9.HRMS(ESI)calcd.for C 29 H 25 F3N6O2547.2064[M+H] + ,found 547.2052.

[0234] Example 35

[0235]

[0236] Following a similar synthetic method as step eight of Example 1, compound HTY-4101 was obtained by reacting acetic acid with compound 10 as a white solid (19 mg, yield 22.61%), using an elution system (DCM:MeOH = 100:1 to 50:1).1 H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.56 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8. 53(d,J=2.0Hz,1H),8.22(s,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.6,2.2Hz ,1H),7.93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.4Hz,1H),7.53(d,J=8.1Hz,1H) ,3.60(s,2H),3.45-3.40(m,4H),2.59(s,3H),2.41-2.32(m,4H),1.99(s,3H). 13 C NMR(151MHz,DMSO-d6)δ168.6,165.2,151.5,151.0,144.2,138.9,138.8,134.3,133.5,132.6,132.1,131.9,131.8,131.0, 130.5,128.6,124.0,122.7,117.7,114.5,112.3,92.4,88.8,57.8,53.4,53.0,46.2,41.4,21.7,20.9.HRMS(ESI)calcd.for C 30 H 27 F3N6O2561.2220[M+H] + ,found 561.2220.

[0237] Example 36

[0238]

[0239] Compound 10 (77 mg, 0.15 mmol), cyanogen bromide (19 mg, 0.18 mmol), and potassium carbonate (32 mg, 0.23 mmol) were weighed into a reaction flask, and N,N-dimethylformamide (3 mL) was added. The reaction was carried out at room temperature for 3 hours. After the reaction was completed, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was evaporated under vacuum, and the mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 60:1) to give a white solid compound HTY-4119 (52 mg, yield 63.82%). 1H NMR(400MHz,DMSO-d6)δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8 .53(d,J=2.0Hz,1H),8.22(s,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.4,2.2 Hz,1H),7.92(dd,J=8.1,1.9Hz,1H),7.71(d,J=8.5Hz,1H),7.53(d,J=8.1Hz ,1H),3.63(s,2H),3.22(t,J=4.9Hz,4H),2.59(s,3H),2.47(d,J=5.0Hz,4H). 13 C NMR (151MHz, DMSO-d6) δ165.2,151.5,151.0,144.2,138.9,134.3,133.5,132.6,131.8,131.8,131.0,130.4,128.6,128. 1,127.9,125.7,123.9,122.7,118.2,114.5,112.3,92.4,88.7,57.8,55.4,51.8,49.1,41.2,20.9.HRMS(ESI)calcd.forC 29 H 24 F3N7O 544.2067[M+H] + ,found 544.2054.

[0240] Example 37

[0241]

[0242] Following a similar synthetic method as step eight of Example 1, compound HTY-4141 was obtained by reacting benzoic acid with compound 10 as a yellow solid (56 mg, yield 59.99%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=1.9Hz,1H),8.53 (d,J=2.0Hz,1H),8.23(d,J=2.5Hz,2H),8.19(d,J=2.0Hz,1H),8.09(d,J=7.5Hz ,1H),7.92(dd,J=8.0,1.9Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.0Hz,1H), 7.44(q,J=3.0Hz,3H),7.41-7.37(m,2H),3.63(s,4H),2.59(s,3H),2.43(s,4H). 13 C NMR (151MHz, DMSO-d6) δ169.4,165.2,151.5,151.0,144.2,138.8,136.4,134.3,133.5,133.3,132.6,132.1,131.8,131.0,130.4,130.0,129. 7,129.0,128.9,128.6,127.4,124.0,123.9,122.7,117.8,114.5,112.3,92.4,88.8,57.8,54.0,53.3,52.9,42.3,20.9.HRMS(ESI)calcd.forC 35 H 29 F3N6O2623.2377[M+H] + ,found 623.2376.

[0243] Example 38

[0244]

[0245] Following a similar synthetic method as in Example 9, compound HTY-4142 was obtained as a yellow solid (17 mg, yield 19.73%) by reacting propionyl chloride with compound 10, using an elution system (DCM:MeOH = 100:1 to 20:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2. 0Hz,1H),8.24-8.21(m,2H),8.20(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.93(dd, J=8.0,2.0Hz,1H),7.75(d,J=8.6Hz,1H),7.53(d,J=8.2Hz,1H),3.60(s,2H),3.44(d,J= 6.9Hz,4H),2.59(s,3H),2.40-2.33(m,4H),2.29(t,J=7.4Hz,2H),0.98(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ171.7,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,12 5.7,124.0,123.9,122.7,117.7,114.5,112.3,92.4,88.8,57.8,53.4,53.0,45.2,41.5,26.0,20.9,9.9.HRMS(ESI)calcd.for C 31 H 29 F3N6O2575.2377[M+H] + ,found 575.2377.

[0246] Example 39

[0247]

[0248] Compound 10 (77 mg, 0.15 mmol), iodoethane (25 mg, 0.16 mmol), and potassium carbonate (32 mg, 0.23 mmol) were weighed into a reaction flask, and N,N-dimethylformamide (3 mL) was added. The reaction was carried out overnight at 40 °C. After the reaction was completed, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed twice with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give a yellow solid compound HTY-4147 (43 mg, yield 52.50%). 1H NMR (400MHz, DMSO-d6) δ13.95 (s, 1H), 10.55 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.53 (d,J=2.0Hz,1H),8.22(d,J=3.7Hz,2H),8.20(d,J=2.0Hz,1H),8.08(dd,J=8.5, 2.3Hz,1H),7.93(dd,J=8.0,2.0Hz,1H),7.71(d,J=8.5Hz,1H),7.53(d,J=8.1Hz ,1H),3.59(s,2H),3.40(s,4H),2.59(s,3H),2.46(s,6H),1.02(t,J=7.2Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ165.2,151.5,151.0,144.2,138.7,134.2,133.5,132.6,131.7,131.0,130.4,128.6,128.0,127.8, 125.7,124.0,123.9,122.7,114.5,112.3,92.4,88.8,57.8,52.5,51.9,31.8,29.5,27.0,22.6,20.9.HRMS(ESI)calcd.for C 30 H 29 F3N6O547.2428[M+H] + ,found 547.2427.

[0249] Example 40

[0250]

[0251] Following a similar synthetic method as in Example 9, compound HTY-4151 was obtained by reacting chloroacetyl chloride with compound 10 as a yellow solid (27 mg, yield 23.91%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.57(s,1H),8.74(d,J=1.9Hz,1H),8.53( d,J=2.0Hz,1H),8.23(s,2H),8.20(d,J=2.0Hz,1H),8.10(dd,J=8.6,2.3Hz,1H),7 .93(dd,J=8.0,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),4.37(s,2 H),3.61(s,2H),3.48(d,J=4.9Hz,4H),2.59(s,3H),2.40(dt,J=23.0,4.9Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ169.1,165.2,165.0,151.5,144.2,138.8,134.3,133.5,132.6,132.0,131.9,131.0,130.5,128.7,128 .0,125.7,124.0,123.9,122.7,117.8,114.5,112.3,92.4,88.8,57.7,53.2,52.7,45.9,42.3,42.2,20.9.HRMS(ESI)calcd.for C 30 H 26 ClF3N6O2595.1831[M+H] + ,found595.1831.

[0252] Example 41

[0253]

[0254] Following a similar synthetic method as in Example 9, compound HTY-4153 was obtained by reacting bromoacetyl bromide with compound 10 as a yellow solid (30 mg, yield 24.74%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94 (s, 1H), 10.59 (s, 1H), 8.74 (d, J = 2.0Hz, 1H), 8.53 (d,J=2.0Hz,1H),8.26-8.22(m,2H),8.20(d,J=2.0Hz,1H),8.13(d,J=8.4Hz,1H) ,7.93(dd,J=8.0,2.0Hz,1H),7.78(d,J=8.6Hz,1H),7.53(d,J=8.1Hz,1H),4.15( s,2H),3.70(s,2H),3.52(s,4H),2.59(s,3H),2.33(s,4H).HRMS(ESI)calcd.for C 30 H 26 BrF3N6O2639.1325[M+H] + ,found639.1328.

[0255] Example 42

[0256]

[0257] Following a similar synthetic method as in Example 39, compound HTY-4154 was obtained by reacting 1-iodobutane with compound 10 as a yellow solid (33 mg, yield 38.31%), using an elution system (DCM:MeOH = 100:1 to 20:1). 1 H NMR (400MHz, DMSO-d6) δ13.95(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2. 0Hz,1H),8.22(d,J=4.4Hz,2H),8.19(d,J=2.0Hz,1H),8.08(dd,J=8.6,2.3Hz,1H),7.93( dd,J=8.0,2.0Hz,1H),7.71(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),3.58(s,2H),3.33(s, 4H), 2.59 (s, 3H), 2.43 (s, 6H), 1.42 (s, 2H), 1.27 (d, J = 7.4Hz, 2H), 0.87 (t, J = 7.3Hz, 3H). 13C NMR(151MHz,DMSO-d6)δ165.2,151.5,144.2,138.7,134.2,133.5,132.6,131.7,131.0,130.4,128.6,128.0,127.8,125.7,124.0 ,123.9,122.7,117.7,114.5,112.3,92.4,88.8,57.8,56.5,55.4,53.0,31.8,30.8,22.6,20.9,20.5,14.3.HRMS(ESI)calcd.forC 32 H 33 F3N6O 575.2741[M+H] + ,found575.2741.

[0258] Example 43

[0259]

[0260] Following a similar synthetic method as in Example 9, compound HTY-4155 was obtained by reacting n-valeryl chloride with compound 10 as a yellow solid (42 mg, yield 36.70%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H),8. 22(t,J=1.9Hz,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.93(dd,J=8.0,2.0Hz,1H), 7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),3.60(s,2H),3.45(d,J=5.3Hz,4H),2.59(s,3H),2.36( d,J=19.0Hz,4H),2.28(t,J=7.5Hz,2H),1.49-1.42(m,2H),1.31-1.26(m,2H),0.87(t,J=7.3Hz,3H). 13C NMR (151MHz, DMSO-d6) δ171.0,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,125.7,12 4.0,123.9,122.7,117.7,114.5,112.3,92.4,88.8,57.8,53.5,53.0,45.4,41.5,32.4,27.5,22.4,20.9,14.3.HRMS(ESI)calcd.for C 33 H 33 F3N6O26O3.2690[M+H] + ,found603.2689.

[0261] Example 44

[0262]

[0263] Following a similar synthetic method as step eight in Example 1, compound HTY-5003 was obtained by reacting n-heptanoic acid with compound 10 as a yellow solid (53 mg, yield 56.06%), using an elution system (DCM:MeOH = 100:1 to 50:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H),8. 22(t,J=2.1Hz,2H),8.19(d,J=2.0Hz,1H),8.09(dd,J=8.5,2.2Hz,1H),7.93(dd,J=8.0,2.0Hz,1H), 7.74(d,J=8.5Hz,1H),7.53(d,J=8.2Hz,1H),3.59(s,2H),3.45(d,J=5.3Hz,4H),2.59(s,3H),2.35( d,J=18.0Hz,4H),2.27(t,J=7.5Hz,2H),1.47(t,J=7.2Hz,2H),1.28-1.24(m,6H),0.87-0.83(m,3H). 13C NMR(151MHz,DMSO-d6)δ171.0,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,125.7,124.0,1 23.9,122.7,117.8,114.5,112.3,92.4,88.8,57.8,53.5,53.1,45.4,41.5,32.7,31.6,28.9,25.3,22.5,20.9,14.4.HRMS(ESI)calcd.for C 35 H 37 F3N6O2631.3003[M+H] + ,found 631.3001.

[0264] Example 45

[0265]

[0266] Following a similar synthetic method as in Example 39, compound HTY-5004 was obtained by reacting 1-iodohexane with compound 10 as a yellow solid (18 mg, yield 13.58%), using an elution system (DCM:MeOH = 100:1 to 30:1). 1 H NMR (400MHz, DMSO-d6) δ13.95(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J =2.0Hz,1H),8.24-8.21(m,2H),8.19(d,J=2.0Hz,1H),8.08(dd,J=8.6,2.2Hz,1H),7 .93(dd,J=7.9,2.0Hz,1H),7.71(d,J=8.5Hz,1H),7.53(d,J=8.2Hz,1H),3.59(s,2H) ,2.59(s,3H),2.33(s,6H),1.44(s,2H),1.25(d,J=4.1Hz,6H),0.85(d,J=7.0Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ165.2,151.5,144.2,138.7,133.5,132.6,131.8,131.0,130.5,128.6,124.0,1 23.9,122.7,117.7,114.5,112.3,92.4,88.8,31.8,31.6,29.5,22.5,20.9,14.4.HRMS(ESI)calcd.for C 34H 37 F3N6O603.3054[M+H] + ,found 603.3046.

[0267] Example 46

[0268]

[0269] Following a similar synthetic method as step eight of Example 1, compound HTY-5005 was obtained by reacting levulinic acid with compound 10 as a yellow solid (69 mg, yield 74.64%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.55(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz, 1H), 8.23 ​​(d, J = 2.7Hz, 2H), 8.20 (d, J = 2.0Hz, 1H), 8.09 (dd, J = 8.6, 2.2Hz, 1H), 7.93 (dd, J = 7. 9,2.0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.2Hz,1H),3.60(s,2H),3.46(q,J=6.1Hz,4H ),2.62(t,J=5.6Hz,2H),2.59(s,3H),2.52(d,J=5.6Hz,2H),2.41-2.32(m,4H),2.10(s,3H). 13 C NMR(151MHz,DMSO-d6)δ207.9,170.1,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,12 7.8,125.7,124.0,122.7,117.8,114.5,112.3,92.4,88.8,57.8,53.3,53.0,45.2,41.7,38.0,30.3,27.1,20.9.HRMS(ESI)calcd.for C 33 H 31 F3N6O3617.2482[M+H] + ,found 617.2479.

[0270] Example 47

[0271]

[0272] The first step, following a similar synthetic method to step eight of Example 1, involved reacting 1,3-dioxolane-2-propionic acid with compound 10 to obtain compound 47-2, a yellow solid (78 mg, yield 69.79%), in an elution system (DCM:MeOH = 100:1 to 30:1). 1 H NMR (400MHz, DMSO-d6) δ13.93(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz ,1H),8.24-8.21(m,2H),8.20(d,J=2.0Hz,1H),8.11-8.07(m,1H),7.93(dd,J=8.0,2.0Hz,1H ),7.75(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),4.82(td,J=4.7,1.2Hz,1H),3.88-3.86(m,2H ),3.75(q,J=1.8Hz,2H),3.60(s,2H),3.45(s,4H),2.59(s,3H),2.38(dd,J=9.4,5.9Hz,4H).

[0273] In the second step, compound 47-2 (70 mg, 0.108 mmol) was weighed into a reaction flask, and 99% formic acid (2 mL) was added. The reaction was carried out overnight at 60 °C. After the reaction was completed, the mixture was quenched with saturated sodium bicarbonate aqueous solution, extracted three times with ethyl acetate, and the organic phases were combined. The mixture was washed twice with saturated sodium chloride, dried with anhydrous sodium sulfate, and the solvent was evaporated under vacuum. The mixture was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 40:1) to obtain a yellow solid HTY-5007 (37 mg, yield 56.89%). 1 HNMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J= 2.0Hz,1H),8.23(t,J=2.2Hz,2H),8.19(d,J=2.0Hz,1H),8.10(dd,J=8.4,2.2Hz,1H),8. 00(s,1H),7.93(dd,J=8.0,2.0Hz,1H),7.76(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),3.6 2(s,2H),3.41-3.36(m,4H),2.59(s,3H),2.51(d,J=2.0Hz,4H),2.38(d,J=23.8Hz,4H). 13CNMR(151MHz,DMSO-d6)δ165.2,161.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.1,131.8,131.0,130.4,128.6,125.7, 124.0,123.9,122.7,117.8,117.8,114.5,112.3,92.4,88.8,57.8,53.7,52.6,45.3,31.8,30.8,29.5,20.9.HRMS(ESI)calcd.forC 32 H 29 F3N6O36O3.2326[M+H] + ,found603.2322.

[0274] Example 48

[0275]

[0276] Following a similar synthetic method as in Example 39, compound HTY-5008 was obtained by reacting 1-iodopropane with compound 10 as a yellow solid (37 mg, yield 44.03%), using an elution system (DCM:MeOH = 100:1 to 15:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz, 1H),8.23(t,J=2.2Hz,2H),8.19(d,J=2.0Hz,1H),8.10(dd,J=8.4,2.2Hz,1H),8.00(s,1H),7 .93(dd,J=8.0,2.0Hz,1H),7.76(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),3.62(s,2H),3.41-3 .36(m,4H),2.59(s,3H),2.51(d,J=2.0Hz,4H),2.38(d,J=23.8Hz,4H).HRMS(ESI)calcd.for C 31 H 31 F3N6O 561.2584[M+H] + ,found 561.2583.

[0277] Example 49

[0278]

[0279] Following a similar synthetic method as step four of Example 16, compound HTY-5010 was obtained by reacting 3-hydroxypropionic acid with compound 10 as a yellow solid (19 mg, yield 21.46%), using an elution system (DCM:MeOH = 100:1 to 20:1). 1 H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.56(s,1H),8.74(d,J=2.0Hz,1H),8.53(d,J=2.0Hz,1H ),8.23(d,J=1.7Hz,2H),8.19(d,J=1.9Hz,1H),8.09(dd,J=8.6,2.2Hz,1H),7.93(dd,J=8.0,2. 0Hz,1H),7.75(d,J=8.5Hz,1H),7.53(d,J=8.2Hz,1H),4.50(t,J=5.4Hz,1H),3.65-3.61(m,2H) ,3.60(s,2H),3.47(t,J=5.0Hz,4H),2.59(s,3H),2.47(t,J=6.7Hz,2H),2.37(d,J=18.6Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ169.8,165.2,151.5,151.0,144.2,138.8,134.3,133.5,132.6,132.2,131.8,131.0,130.5,130.1,128 .6,128.0,125.7,124.0,122.7,114.5,112.3,92.4,88.8,58.0,57.8,53.5,53.0,45.6,41.4,36.2,20.9.HRMS(ESI)calcd.for C 31 H 29 F3N6O3591.2326[M+H] + ,found 591.2325.

[0280] Example 50

[0281]

[0282] Following a similar synthetic method as in Example 39, compound HTY-5011 was obtained by reacting 2-iodopropane with compound 10 as a yellow solid (37 mg, yield 44.03%), using an elution system (DCM:MeOH = 100:1 to 10:1). 1H NMR(400MHz,DMSO-d6)δ13.95(s,1H),10.57(s,1H),8.74(d,J=2.0Hz,1H),8 .53(d,J=2.0Hz,1H),8.23(s,2H),8.20(d,J=1.9Hz,1H),8.09(dd,J=8.4,2.2 Hz,1H),7.93(dd,J=8.0,2.0Hz,1H),7.73(d,J=8.5Hz,1H),7.53(d,J=8.2Hz ,1H),3.60(s,2H),2.94-2.60(m,5H),2.59(s,3H),2.33(s,4H),1.08(s,6H). 13 C NMR(151MHz,DMSO-d6)δ165.2,151.5,151.0,144.2,138.8,134.2,133.5,132.6,131.8,131.7,131.0,130.4,128.6,128.0,127.8,125 .7,124.0,123.9,122.7,117.8,117.7,114.5,112.3,92.4,88.8,60.2,57.5,48.3,29.5,29.5,24.4,22.6,20.9.HRMS(ESI)calcd.for C 31 H 31 F3N6O 561.2584[M+H] + ,found561.2574.

[0283] Example 51

[0284]

[0285] Following a similar synthetic method as in Example 39, compound HTY-5012 was obtained by reacting benzyl bromide with compound 10 as a white solid (15 mg, yield 16.48%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ13.94(s,1H),10.54(s,1H),8.74(s,1H),8.53(s,1H),8.23(s,1H),8.20(dd,J=8.0,2.1Hz,2H),8.07(d,J=8.6Hz,1H),7.92 (d,J=7.1Hz,1H),7.71(d,J=8.5Hz,1H),7.53(d,J=8.1Hz,1H),7.34-7.22 (m,5H),3.57(s,2H),3.47(s,2H),3.32(s,4H),2.59(s,3H),2.41(s,4H). 13 C NMR(151MHz,DMSO-d6)δ165.2,151.5,151.0,144.2,138.7,134.3,133.5,132.7,131.7,131.0,130.4,129.3,128.6,12 5.7,124.0,123.9,122.7,117.7,117.7,114.5,112.3,92.4,88.8,62.5,57.9,53.3,53.1,20.9.HRMS(ESI)calcd.forC 35 H 31 F3N6O 609.2584[M+H] + ,found 609.2573.

[0286] Example 52

[0287]

[0288] Following a similar synthetic method as step four of Example 16, compound HTY-5013 was obtained by reacting acetic acid and compound 14 as a yellow solid (18 mg, yield 21.42%), using an elution system (DCM:MeOH = 100:1 to 40:1). 1H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.73(dd,J=4.5,1.6Hz,1H),8.27(dd,J=9.3,1.6Hz,1H ),8.24(s,1H),8.22(dd,J=4.6,2.0Hz,2H),8.09(d,J=8.1Hz,1H),7.95(dd,J=8.0,2.0Hz,1H ),7.75(d,J=8.6Hz,1H),7.56(d,J=8.1Hz,1H),7.40(dd,J=9.2,4.4Hz,1H),3.60(s,2H),3.4 6-3.41(m,4H),2.61(s,3H),2.36(dd,J=24.1,5.4Hz,4H),1.99(s,3H).HRMS(ESI)calcd.for C 30 H 27 F3N6O2561.2220[M+H] + ,found 561.2217.

[0289] Example 53

[0290]

[0291] Following a similar synthetic method as in Example 39, compound HTY-5014 was obtained by reacting iodoethane with compound 14 as a yellow solid (33 mg, yield 40.27%), using an elution system (DCM:MeOH = 100:1 to 10:1). 1 H NMR (400MHz, DMSO) δ10.57(s,1H),8.73(dd,J=4.5,1.6Hz,1H),8.26(dd,J=9.3,1.6Hz,1H) ,8.23(s,1H),8.22(t,J=2.0Hz,2H),8.08(dd,J=8.5,2.3Hz,1H),7.95(dd,J=8.0,2.0Hz,1 H),7.71(d,J=8.6Hz,1H),7.55(d,J=8.1Hz,1H),7.39(dd,J=9.2,4.5Hz,1H),3.58(s,2H), 2.61(s,3H),2.56-2.51(m,2H),2.43(s,8H),1.01(t,J=7.2Hz,3H).HRMS(ESI)calcd.forC 30 H 29 F3N6O 547.2428[M+H] + ,found 547.2418.

[0292] Example 54

[0293]

[0294] In the first step, compound 54-1 (300 mg, 0.69 mmol) was dissolved in DMF (3 mL), followed by the addition of potassium carbonate (95 mg, 0.69 mmol) and tert-butylpiperazine carboxylate (130 mg, 0.69 mmol). The reaction was carried out at 70 °C for 5 hours. After the reaction was completed, the reaction mixture was quenched with water and filtered to obtain a solid, which could be used directly in the next step without purification, yielding a yellow solid 54-2 (369 mg, yield 92.25%). 1 H NMR (400MHz, DMSO-d6) δ10.17(s,1H),9.27(d,J=2.2Hz,1H),8.97(s,1H),8.68(dd,J=4.8,1. 7Hz,1H),8.51(d,J=5.1Hz,1H),8.47(dt,J=8.1,2.0Hz,1H),8.08(d,J=2.2Hz,1H),7.91(d,J =8.2Hz,2H),7.52(dd,J=8.0,4.8Hz,1H),7.48(dd,J=8.2,2.2Hz,1H),7.46-7.39(m,3H),7.2 0(d,J=8.3Hz,1H),3.55(s,2H),3.31(s,4H),2.33(t,J=5.1Hz,4H),2.22(s,3H),1.39(s,9H).

[0295] In the second step, compound 54-2 (369 mg, 0.63 mmol) was weighed into a reaction flask, and 3 mL of 1,4-dioxane solution in 4 M hydrochloric acid was added. The reaction was carried out at room temperature for 4 hours. After the reaction was completed, the solvent was evaporated under vacuum. No purification was required, and the solvent was used directly in the next step to obtain compound 54-3 (287 mg, 95.23%).

[0296] The third step involved reacting iodoethane and compound 54-3 with a synthetic method similar to that in Example 39 to obtain compound HTY-5018, a yellow solid (15 mg, yield 19.23%), as a yellow solid, in an elution system (DCM:MeOH = 20:1 to 5:1). 1HNMR(400MHz,DMSO-d6)δ10.18(s,1H),9.27(d,J=2.2Hz,1H),8.97(s,1H),8.68(dd,J=4.8,1 .7Hz,1H),8.51(d,J=5.1Hz,1H),8.47(dt,J=8.1,2.0Hz,1H),8.08(d,J=2.2Hz,1H),7.92(d,J =8.0Hz,2H),7.52(dd,J=8.0,4.7Hz,1H),7.48(dd,J=8.2,2.2Hz,1H),7.43(dd,J=6.8,5.4Hz ,3H),7.20(d,J=8.3Hz,1H),3.56(s,2H),3.31(s,4H),2.33(s,6H),2.22(s,3H),1.03(s,3H). 13 C NMR(151MHz,DMSO-d6)δ165.7,162.1,161.7,160.0,151.9,148.7,138.3,137.7,134.9,134.3,132.7, 130.5,130.1,129.1,128.1,124.3,117.7,117.2,108.0,61.8,52.4,51.8,18.1.HRMS(ESI)calcd.for C 30 H 33 N7O 508.2819[M+H] + ,found 508.2802.

[0297] Example 55

[0298]

[0299] Following a similar synthetic method as in Example 9, compound HTY-5021 was obtained by reacting acetyl chloride with compound 54-3 as a yellow solid (20 mg, yield 25.58%), using an elution system (DCM:MeOH = 50:1 to DCM:MeOH:TEA 8:1:0.09). 1H NMR (400MHz, DMSO-d6) δ10.18(s,1H),9.27(dd,J=2.3,0.9Hz,1H),8.97(s,1H),8.68(d d,J=4.8,1.7Hz,1H),8.51(d,J=5.2Hz,1H),8.49-8.45(m,1H),8.08(d,J=2.2Hz,1H),7 .92(d,J=8.1Hz,2H),7.54-7.48(m,2H),7.44(dd,J=11.4,6.5Hz,3H),7.20(d,J=8.3Hz ,1H),3.57(s,2H),3.45-3.40(m,4H),2.35(d,J=26.9Hz,4H),2.22(s,3H),1.97(s,3H). 13 CNMR(151MHz,DMSO-d6)δ168.6,165.7,162.1,161.7,160.0,151.9,148.7,142.1,138.3,137.7,134.9,134.4,132. 7,130.5,129.2,128.1,124.3,117.7,117.2,108.0,61.9,53.3,52.8,46.1,41.3,21.7,18.1.HRMS(ESI)calcd.for C 30 H 31 N7O2522.2612[M+H] + ,found522.2598.

[0300] Example 56

[0301]

[0302] The first step, following a similar synthetic method to step two of Example 1, involved reacting N-ethylpiperazine and compound 2 to obtain compound 56-1, a yellow solid (300 mg, yield 77.40%), in an elution system (PE:EA = 5:1 to 1:1). 1 H NMR (400MHz, CDCl3) δ8.49 (d, J=2.4Hz, 1H), 8.35 (dd, J=8.6, 2.4Hz, 1H), 8.09 (d, J= 8.6Hz,1H),3.74(s,2H),2.55(s,8H),2.45(d,J=7.2Hz,2H),1.10(t,J=7.2Hz,3H).

[0303] In the second step, following a similar synthesis method as in step three of Example 1, compound 56-2 was obtained as a yellow solid (478 mg, yield 45.23%), using an elution system (DCM:MeOH = 50:1 to 20:1).

[0304] The third step involves a synthesis method similar to step six of Example 1, where methyl 3-iodo-4-methoxybenzoate and compound 56-2 are reacted to yield compound 56-3, which is a white solid (500 mg, yield 88.21%), in an elution system (DCM:MeOH = 50:1 to 30:1). 1 HNMR (400MHz, DMSO) δ10.39(s,1H),8.42(d,J=2.2Hz,1H),8.17(d,J=2.2Hz,1H),8.03(dt,J=8.5,2.4Hz,2H),7. 70(d,J=8.6Hz,1H),7.15(d,J=8.8Hz,1H),3.92(s,3H),3.56(s,2H),2.48-2.20(m,10H),0.98(t,J=7.2Hz,3H).

[0305] Fourth step, following a similar synthesis method as in step four of Example 1, compound 56-4 was obtained as a yellow solid (210 mg, yield 86.13%), using an elution system (DCM:MeOH = 100:1 to 50:1). 1 HNMR(400MHz,DMSO)δ10.44-10.35(m,1H),8.19(s,1H),8.11(s,1H),8.06-7.98(m,2H),7.69(t,J=7.1Hz,1H),7 .23(d,J=7.3Hz,1H),4.36(d,J=5.1Hz,1H),3.96-3.84(m,3H),3.56(s,2H),2.41(s,10H),0.99(d,J=6.6Hz,3H).

[0306] Fifth step, following a similar synthetic method to step five of Example 1, compound 56-4 was reacted with 5-bromo-1H-pyrazolopyridine to obtain compound HTY-5038, which was a yellow solid (70 mg, yield 27.43%), in an elution system (DCM:MeOH = 50:1 to 15:1). 1H NMR (400MHz, DMSO-d6) δ13.93(s,1H),10.45(s,1H),8.68(d,J=2.0Hz,1H),8.47(d,J=2.0Hz,1H),8.23-8.20(m,3H),8.06(dt,J=8.8 ,2.0Hz,2H),7.70(d,J=8.5Hz,1H),7.28(d,J=8.9Hz,1H),3.97(s,3H),3.57(s,2H),2.39(d,J=21.5Hz,11H),0.99(d,J=7.2Hz,3H).

[0307] Example 57

[0308]

[0309] The first step, following a similar synthetic method to step five of Example 1, involved reacting compound 57-1 with 5-bromo-1H-pyrazolopyridine to obtain compound 57-2, which was a yellow solid (481 mg, yield 25.32%), in an elution system (PE:EA = 100:1 to 5:1).

[0310] In the second step, following a similar synthesis method as step six of Example 1, compound HTY-5039 was obtained by reacting compound 56-2 and compound 57-2 as a yellow solid (88 mg, yield 47.16%), using an elution system (DCM:MeOH = 50:1 to 30:1). 1 H NMR (400MHz, DMSO-d6) δ13.96(s,1H),10.64(s,1H),8.73(d,J=2.0Hz,1H),8.52(d,J=2.0Hz,1H),8.24-8.21(m,3H),8.08(dd,J=8.5,2.2Hz,1H), 8.03-8.00(m,1H),7.84-7.81(m,1H),7.73(d,J=8.6Hz,1H),7.64(t,J=7 .8Hz,1H),3.59(s,2H),3.17(s,2H),2.46(s,9H),1.02(t,J=7.1Hz,3H).

[0311] Example 58 Compound on Ba / F3 Bcr-Abl T315I Cell proliferation inhibition activity

[0312] Cell viability was determined using the CCK-8 assay. 8000 Ba / F3 Bcr-Abl cells were... T315ICells were seeded in each well of a 96-well plate and incubated in a humidified incubator for 24 hours to allow adhesion. Compounds (0 μM–100 μM) were dissolved in DMSO and added to each well, maintaining a final DMSO concentration of 0.1%. After 72 hours of incubation, 10 μL of CCK-8 reagent (CCK-8, DOJINDO, #CK04) was added to each well and incubated for 2 hours. The absorbance was then measured at 450 nm and 650 nm using a plate reader (Synergy HI, BioTek Instruments, Inc., Vermont, US). Cell viability (VR) was calculated as (A-A0) / (As-A0) × 100%, where A is the absorbance of the experimental group, As is the absorbance of the control group (DMSO as a control), and A0 is the absorbance of the blank group (cell-free group). Finally, the IC50 was calculated using GraphPad Prism software. 50 value.

[0313] The test results are shown in Table 1: The compounds of the present invention affect Ba / F3 Bcr-Abl T315I The cells have strong inhibitory activity against cell proliferation.

[0314] Table 1. Effects of compounds on Ba / F3 Bcr-Abl T315I Cell proliferation inhibition activity

[0315] Compound Number <![CDATA[IC 50 (nM)]]> Compound Number <![CDATA[IC 50 (nM)]]> Compound Number <![CDATA[IC 50 (nM)]]> Bal-6 54.5 HTY-4096 76.6 HTY-5010 23.39 Bal-7 1.425 HTY-4093 94.3 HTY-4155 21.62 Bal-8 19.5 HTY-4095 31.3 HTY-5003 70.31 Bal-9 50.2 HTY-4103 16.5 HTY-4147 3.65 Bal-10 87.8 HTY-4100 24.3 HTY-5008 1.72 Bal-11 79.9 HTY-4119 37.9 HTY-4154 14.03 Bal-12 849.9 HTY-4141 181.1 HTY-5004 14.03 Bal-22 63.6 HTY-4101 8.53 HTY-5011 14.33 HTY-4015 117.4 HTY-4111 15.5 HTY-5012 35.98 HTY-4019 86.9 HTY-4142 23.2 HTY-4045 381.4 ​ 767.3 ​ 22.07 ​ 58.4 ​ 844.8 ​ 19.61 ​ 232.0 ​ 61.9 ​ 7.99 ​ 463.4 ​ 82.2 ​ 5.38 ​ 216.4 ​ 152.8 ​ 25.1 ​ 23.42 ​ 95.3 ​ 14.1 ​ 87.8 ​ 59.5 ​ 13.7 ​ ﹥1000 ​ 87.2 ​ 22.34 ​ ﹥1000 ​ 11.75 ​ 5.144

[0316] Example 59 tests the compounds of the present invention in Ba / F3 Bcr-Abl T315I Cellular degradation activity against T315I mutant

[0317] Take 2×10 6 Ba / F3 Bcr-Abl T315I Cells were incubated with the compound (1-100 nM) at 37°C for 24 hours in complete culture medium. Cells were collected by centrifugation, washed twice with cold phosphate-buffered saline (PBS), and centrifuged again to obtain a cell pellet. The cell pellet was then washed with PBS and irrigated with RIPA (Thermo Scientific) containing 2.5% protease and phosphatase inhibitors (Beyotime, P1046). TM The protein was lysed (#89900). Protein concentration was determined using a BCA protein assay kit (Beyotime, P0009) and a Synergy H1 hybrid microplate reader (BioTek).

[0318] Samples were dissolved in SDS-polyacrylamide gel and transferred to a polymer membrane. The membrane was then blocked for 1 hour at room temperature with 5% bovine serum albumin (BSA) in TBST (0.1% Tween in Tris buffer). After blocking, the membrane was incubated overnight at 4°C with the corresponding primary antibody. After incubation, the membrane was washed with TBST (4 × 10 min) and then incubated with a suitable secondary antibody. Finally, the blot was washed again with TBST, and the blot was developed using the Super Signal Sidurah kit (Thermo Scientific) and imaged using an Amersham Imager 600 system (GE Healthcare). The half-degradation target protein Bcr-Abl was calculated. T315I concentration (DC) 50 ).

[0319] The results are as follows ​ As shown in Table 2.

[0320] Table 2 shows the compounds in Ba / F3 Bcr-Abl. T315I Cellular degradation activity against T315I mutant

[0321] ​ <![CDATA[DC 50 (nM)]]> ​ <![CDATA[DC 50 (nM)]]> ​ <![CDATA[DC 50 (nM)]]> ​ ​ ​ 57.5 ​ 16.18 ​ 14.2 ​ 21.2 ​ 27.5 ​ 124.9 ​ 27.4 ​ 165.9 ​ 56.8 ​ 21.9 ​ 46.6 ​ 20.3 ​ 20.3 ​ 15.2

[0322] The results showed that the compound prepared in this invention was in Ba / F3 Bcr-Abl T315I The cells exhibited strong degradation activity against the T315I mutant, while the compound GDZ824 showed no degradation activity.

[0323] Example 60: Degradation activity of compounds HTY-4147 and HTY-5018 against BCR-ABL protein in K562 cells.

[0324] Take 2×10 6 K562 cells were incubated with the compound (6.25-200 nM) at 37°C for 24 hours in complete culture medium. Cells were collected by centrifugation, washed twice with cold phosphate-buffered saline (PBS), and centrifuged again to obtain a cell pellet. The cell pellet was then washed with PBS and irrigated with RIPA (ThermoScientific Acid-Resistant Agent) containing 2.5% protease and phosphatase inhibitor (Beyotime, P1046). TM The protein was lysed (#89900). Protein concentration was determined using a BCA protein assay kit (Beyotime, P0009) and a Synergy H1 hybrid microplate reader (BioTek).

[0325] Samples were dissolved in SDS-polyacrylamide gel and transferred to a polymer membrane. The membrane was then blocked for 1 hour at room temperature with 5% bovine serum albumin (BSA) in TBST (0.1% Tween in Tris buffer). After blocking, the membrane was incubated overnight at 4°C with the appropriate primary antibody. After incubation, the membrane was washed with TBST (4 × 10 min) and then incubated with a suitable secondary antibody. Finally, the blot was washed again with TBST, and the image was developed using the Super Signal Sidurah kit (Thermo Scientific). Imaging results were obtained using an Amersham Imager 600 system (GE Healthcare). ​ As shown, HTY-4147 and HTY-5018 exhibit potent degradation activity against Bcr-Abl protein in K562 cells, DC 50 The values ​​are 24.4 nM and 105.68 nM, respectively.

[0326] Example 61: Inhibitory activity of compound HTY-4147 on the proliferation of different tumor cells

[0327] The cell lines used in this experiment were MV4-11 (human acute myeloid leukemia cells), K562 (human chronic myeloid leukemia cells), HCT116 (human colorectal adenocarcinoma cells), SKBR3 (human breast cancer cells), MOLT4 (human acute lymphoblastic leukemia cells), ASPC1 (human pancreatic cancer cells), JURKAT (human acute myeloid leukemia cells), BaF3-Bcr-Abl-T315I (a model cell line stably expressing the Bcr-Abl fusion protein T315I drug-resistant mutant), and MRC-5 (human lung fibroblasts), which were obtained from ATCC and the Shanghai Cell Bank, respectively. 3000-10000 cells / well were seeded into 96-well plates and incubated overnight. Then, different concentrations of compounds (0-30 μM) were added for continuous treatment for 72 hours. CCK8 reagent was then added, and incubation continued for 1-3 hours. The absorbance at 450 nm and 650 nm was then measured using a high-performance microplate reader. Its half-maximal inhibitory concentration (IC50) was calculated using GrapPad Prism 5.0 software. 50 ).

[0328] The results (Tables 3 and 4) showed that the compounds of this invention exhibited strong inhibitory activity against K562 and other cell lines, especially compound HTY-4147, which demonstrated excellent resistance-overcoming activity, cell selectivity, and safety. Compound HTY-4147 showed slightly better inhibitory activity against BaF3-Bcr-Abl-T315I cells than GZD824, and significantly better than marketed drugs dasatinib and imatinib. Its selectivity against non-ABL-driven tumor cells such as ASPC1, MOLT4, and HCT116 was superior to GZD824. Its toxicity to MRC-5 cells was significantly lower than that of GZD824, indicating that compounds such as HTY-4147 of this invention have better safety compared to GZD824.

[0329] Table 3. Inhibitory activity of compounds against the proliferation of different tumor cells.

[0330]

[0331] Table 4. Inhibitory activity of compounds against the proliferation of different tumor cells.

[0332]

[0333]

[0334] Pharmacokinetics of compound HTY-4147 in Example 62

[0335] The pharmacokinetics of compound HTY-4147 (2 mg / kg) were systematically evaluated in a rat model. Six male SPF-grade SD rats were randomly divided into two groups according to body weight: an intravenous injection group (n=3) and a gavage group (n=3), with doses of 2 mg / kg and 10 mg / kg, respectively. Blood samples were collected from the orbital venous plexus in both the intravenous and oral administration groups at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. The blood samples were temporarily stored in an ice box, and plasma was separated by centrifugation at 4000 rpm for 10 min at 4 °C. The plasma drug concentration was determined using an established LC-MS method. The plasma drug concentration data were processed using Phoenix WinNonlin 8.1, and the main pharmacokinetic parameters were calculated using a non-compartmental model.

[0336] The results show ( ​Following intravenous administration, moderate systemic clearance (CLz = 45.00 ± 10.45 mL / min / kg), extensive tissue distribution (Vz = 17256 ± 4944 mL / kg), and an elimination half-life of 4.41 ± 0.39 hours were observed. Oral bioavailability reached 69.78% ± 6.54%. After oral gavage, the compound exhibited consistent absorption (Tmax = 4.00 hours) and peak plasma concentration (Cmax = 138.81 ± 24 ng / mL), with an oral elimination half-life (4.36 hours) very close to that of intravenous administration (4.41 hours). High bioavailability and low pharmacokinetic variability (SD < 15%) provide a solid foundation for druggability and have the potential to become a carrier of Bcr-Abl and Bcr-Abl. T315I These are candidate oral drugs for tumors and have potential for clinical application.

[0337] Example 63: Inhibitory effect of compound HTY-4147 on BaF3-BCR-ABL-T315I xenografts in vivo.

[0338] Animal experiments were approved by the institution's animal ethics committee. CB17-SCID mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. After quarantine, they were subcutaneously injected with 2×10⁻⁶ mice. 6 BaF3-BCR-ABL-T315I cells were used until the tumor volume reached 100-200 mm. 3 Animals were then randomly assigned to groups and administered the test substance HTY-4147 (10 mg / kg, qd; 20 mg / kg, qod), the positive control drug (GZD824, 20 mg / kg, qod), or the solvent (0.5% CMC-Na, 0.1 ml / 10 g) by gavage. Animal weight and tumor size were measured every other day. Tumor volume (V) was calculated as follows: V = L × W × W / 2 (L, tumor length; W, tumor width). Animals were sacrificed on day 15, and tissues were collected for further analysis. The tumor growth inhibition value (TGI) was calculated using the following formula: TGI = (1 - V) / 2 给药 / V 溶媒 )×100%,(V 给药 V represents the average tumor volume in the treated group of animals; 溶媒 (This represents the average tumor volume in the solvent group animals.)

[0339] The results show ( ​Compound HTY-4147, administered once daily at a dose of 10 mg / kg for 13 consecutive days, showed a tumor inhibition rate of 32.7%; administered every two days at a dose of 20 mg / kg for 13 consecutive days, the tumor inhibition rate reached 67.87%. Both results were better than the tumor inhibition rate (26.2%) of the control drug GZD824 (unsalted) in the same batch of experiments, administered every two days at a dose of 20 mg / kg for 13 consecutive days. During the treatment period, no significant weight loss, death, or other obvious toxic side effects were observed in any of the treatment groups.

[0340] Example 64: Inhibition of cell cycle and apoptosis of BaF3-BCR-ABL-T315I by compound HTY-4147

[0341] To evaluate the effect of compound 4147 on Ba / F3 Bcr-Abl T315I The effects of cell cycle distribution and apoptosis were investigated by placing target cells in the logarithmic growth phase at an appropriate density (5 × 10⁻⁶). 5 Cells (per well) were seeded in 6-well plates and cultured overnight in medium containing 10% FBS. The next day, the medium was replaced with fresh medium containing different concentrations of the compound (dissolved in an appropriate solvent, such as DMSO, final concentration ≤0.1%) or a solvent control, and incubated for the set time (24 or 48 hours). After treatment, the culture supernatant and trypsin-digested adherent cells were collected, combined, and centrifuged (300×g, 5 min, 4℃), and washed once with PBS.

[0342] For cell cycle analysis: Cell pellets were fixed overnight at -20°C with pre-cooled 70% ethanol; after washing with PBS, they were resuspended in PBS containing RNase A (50-100 μg / mL) and incubated at 37°C for 30 min to degrade RNA; propidium iodide (PI, final concentration 50 μg / mL) was added, and staining was performed at 4°C in the dark for 15-30 min; DNA content was detected by flow cytometry (FL2 / FL3 channels), and the proportion of cells in G0 / G1, S, and G2 / M phases was analyzed.

[0343] For apoptosis detection: Cell pellet was resuspended directly in 100 μL Annexin V binding buffer, Annexin V-FITC (according to the manufacturer's instructions) and PI (final concentration 50 μg / mL) were added, and the mixture was incubated at room temperature in the dark for 15 min; then 400 μL Annexin V binding buffer was added, and flow cytometry analysis was immediately performed (FITC: FL1 channel; PI: FL2 / FL3 channel) to distinguish between live cells (Annexin V- / PI-) and early apoptotic cells (Annexin V- / PI-). + / PI-), late apoptotic / necrotic cells (Annexin V) + / PI +The result is as follows: ​ As shown, compound 4147 is effective against Ba / F3 Bcr-Abl. T315I It has a significant impact on cell cycle and apoptosis, causing cell cycle arrest and inducing apoptosis.

[0344] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. Aryl amide compounds having the structure shown in Formula I, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, Formula I in, R1 is selected from: , ; R4 is selected from: cyano; R5 is selected from: C3-C6 cycloalkyl groups; R6 is selected from: one or more R... 10 Substituted C1-C3 alkyl groups, 3-8 membered heterocyclic groups, , , Each R 10 Each is independently selected from: halogen and formaldehyde groups; R is selected from: hydrogen, C1-C6 alkyl, C1-C6 alkoxy.

2. The aryl amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R5 is selected from: cyclopropyl.

3. The aryl amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R6 is selected from: one or more R... 10 Substituted C1-C3 alkyl groups , , , Each R 10 Each group is independently selected from: fluorine, chlorine, and formaldehyde groups.

4. The aryl amide compound according to claim 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R6 is selected from: , , , Chlorofluoromethyl, 2-formaldehyde ethyl.

5. The aryl amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R1 is selected from: 、 、 、 、 、 、 。 6. The aryl amide compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, R is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy.

7. The aryl amide compound according to claim 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R is selected from: hydrogen, methyl, ethyl, methoxy, ethoxy.

8. The aryl amide compound according to claim 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R is selected from: hydrogen, methyl.

9. The aryl amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The aryl amide compounds are selected from the following compounds: 。 10. The use of the aryl amide compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a Bcr-Abl protein degrading agent.

11. The use of the aryl amide compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a mutant Bcr-Abl protein degrader.

12. The application according to claim 11, characterized in that, The mutations include: T315I mutation, M244V mutation, G250E mutation, Q252H mutation, H396P mutation, Y253F mutation, E255K mutation, and F317L mutation.

13. The application according to claim 12, characterized in that, The mutation is the T315I mutation.

14. The use of the arylamide compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for treating and / or preventing tumors, wherein the tumor is a tumor carrying the Bcr-Abl fusion gene.

15. The application according to claim 14, characterized in that, The tumor is a tumor carrying a mutated Bcr-Abl gene.

16. The application according to claim 15, characterized in that, The mutations include: T315I mutation, M244V mutation, G250E mutation, Q252H mutation, H396P mutation, Y253F mutation, E255K mutation, and F317L mutation.

17. The application according to claim 16, characterized in that, The mutation is the T315I mutation.

18. The application according to claim 14, characterized in that, The tumors mentioned are leukemia, colorectal adenocarcinoma, breast cancer, and pancreatic cancer.

19. The application according to claim 18, characterized in that, The tumor is leukemia, specifically acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, or acute lymphoblastic leukemia.

20. The application according to claim 19, characterized in that, The tumor cells of the leukemia are MV4-11 cells and K562 cells.

21. A pharmaceutical composition for the prevention or treatment of tumors, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises an aryl amide compound as described in any one of claims 1-9 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

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