Pyrimidine diamine compounds and uses thereof

Abstract

The application provides a pyrimidine diamine compound with a structure shown in formula I or a pharmaceutically acceptable salt or a stereoisomer thereof, and an application thereof. The pyrimidine diamine compound provided by the application is a molecular glue degrader for ALK and FAK and other kinases, can effectively degrade wild-type ALK and FAK kinases and mutant ALK kinases, has a good antiproliferation effect on tumors caused by high expression of ALK or abnormal activation of ALK, has good selectivity, small toxic and side effects, and good safety. The pyrimidine diamine compound of the application can reduce the activity of ALK and FAK proteins from the source, compared with traditional inhibitors, can significantly reduce the occurrence rate of drug resistance mutations, and effectively overcome drug resistance caused by existing drugs. Formula I

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to antitumor drugs, and more particularly to pyrimidine diamine compounds and their application as molecular glues for ALK and FAK. Background Technology

[0002] Lung cancer is the most common malignant tumor worldwide and the leading cause of cancer death. Non-small cell lung cancer (NSCLC) accounts for 85% to 90% of all lung cancers. Although its survival rate has improved with advancements in targeted therapy, immunotherapy, and other therapies, the overall 5-year survival rate is still less than 30%. Therefore, there is an urgent need to develop new drugs to meet the clinical treatment needs of NSCLC.

[0003] Anaplastic lymphoma kinase (ALK) belongs to the insulin receptor superfamily and is mainly composed of an extracellular domain, a transmembrane domain, and an intracellular kinase domain. ALK is expressed at low levels in normal human tissues, and its aberrant activation is a key driver in tumors such as non-small cell lung cancer (NSCLC). It exists in various abnormal forms, including fusion, point mutations, and amplification, with fusion mutations being the most prevalent type in NSCLC. In 2007, researchers first identified the EML4-ALK fusion gene in NSCLC patient samples, and subsequently discovered other ALK fusion forms such as KIF5B-ALK, TFG-ALK, and CD74-ALK. ALK fusion proteins can induce ligand-independent dimerization, leading to autophosphorylation of the intracellular tyrosine kinase domain and continuous activation of downstream signaling pathways such as PI3K-AKT and MAPK / ERK, driving malignant proliferation of tumor cells. Besides fusion mutations, ALK point mutations and amplification are also important forms of aberrant activation.

[0004] Currently, the FDA has approved third-generation ALK inhibitors for ALK-positive NSCLC, while fourth-generation inhibitors are in clinical development. First-generation inhibitors, represented by crizotinib, have some selectivity for wild-type ALK and common fusion mutants, but exhibit cross-inhibition against ROS1 and MET, and most patients develop resistance after about 10 months of treatment. The main resistance mechanisms are L1196M and G1269A mutations in the ALK kinase domain, as well as ALK amplification and bypass activation (EGFR / MET). Second-generation inhibitors, including ceritinib, alectinib, brigatinib, and ensartinib, significantly improve ALK inhibitory activity and selectivity, and reduce cross-inhibition against other kinases. Cross-linking inhibition can effectively overcome common mutation resistance to crizotinib, but second-generation drugs can still lead to secondary resistance, especially the highly resistant ALK mutation G1202R, followed by F1174C / L and I1171N. Third-generation drugs, represented by lorlatinib, have broad-spectrum selectivity for the ALK kinase domain and can effectively overcome first / second-generation resistance mutations (including G1202R), but clinical application still faces the challenge of complex resistance mutations (such as G1202R+L1196M and G1202R+F1174C). Multiple studies have shown that, in addition to target protein mutation resistance, compensatory activation and upregulation of FAK are also key bypass mechanisms for acquired resistance to ALK inhibitors. Although several cases of FAK inhibitors overcoming ALK inhibitor resistance have been reported, the non-enzymatic function of FAK may play a more important role in resistance; therefore, new drugs based on novel strategies are needed to overcome ALK inhibitor resistance.

[0005] Targeted protein degradation (TPD) utilizes endogenous cellular degradation systems to selectively induce the ubiquitination and degradation of target proteins, effectively inhibiting both enzymatic and non-enzymatic functions. It shows broad application prospects in areas such as tumor therapy and overcoming drug resistance. Protein degradation-targeting chimeras (PROTACs) and molecular glue degraders are currently the two most promising targeted protein degradation technologies. PROTACs can simultaneously bind to the target protein and E3 ligase, forming a ternary complex, thereby inducing ubiquitination modification of the target protein, which is ultimately completely degraded by the proteasome. Several studies have reported the design, synthesis, and successful acquisition of multiple ALK-targeting PROTACs using ceritinib as an ALK ligand and pomalidomide as an E3 ligand, achieving potent ALK protein degradation. However, PROTACs have large molecular weights and poor physicochemical properties, making oral administration difficult and resulting in poor drug efficacy. Molecular glue degraders are a class of small molecule compounds that do not require linkers and have smaller molecular weights. They often have better pharmacokinetic properties and cell permeability, but their molecular design is difficult. Currently, there are still few reports on molecular glue degraders targeting ALK and ALK / FAK dual targets. Summary of the Invention

[0006] Based on this, the purpose of this invention is to provide a molecular glue degrader for ALK and FAK, which can effectively degrade ALK and FAK kinases, thereby effectively inhibiting the kinase activity of ALK and its mutants, and has a good anti-proliferative effect on tumors caused by high ALK expression or abnormal ALK activation.

[0007] The technical solutions for achieving the above objectives include the following.

[0008] In a first aspect, the present invention provides pyrimidine diamine compounds having the structure shown in Formula I, or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0009]

[0010] Formula I

[0011] Wherein, R is selected from: R1 substituted or unsubstituted 5-7 membered heterocyclic group, R2 substituted or unsubstituted C5-C7 cycloalkyl group, 5-6 membered heteroaryl group, and C1-C3 alkyl substituted 5-6 membered heteroaryl group;

[0012] R1 is selected from: , C1-C6 alkyl, deuterated C1-C6 alkyl, cyano;

[0013] R2 is selected from: C1-C6 alkyl groups, halogens;

[0014] R3 is selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, 3-4 membered heterocyclic group, 5-10 membered heteroaryl group. , , , , , R6 and R7 are independently selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, cyano, C3-C6 cycloalkyl, R9-substituted benzoyl, and R9-substituted naphthoyl; R9 is selected from: hydrogen, C1-C6 alkoxy, C1-C6 alkyl, and halogen; R8 is selected from: hydrogen and C1-C6 alkyl.

[0015] R4 is selected from: C1-C6 alkoxy, C3-C6 cycloalkyloxy, halogen-substituted C1-C6 alkoxy, and halogen-substituted C3-C6 cycloalkyloxy.

[0016] R5 is selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl. .

[0017] In some embodiments, R4 is selected from: C1-C3 alkoxy, C3-C4 cycloalkyloxy, ... , Halogen-substituted C3-C4 cycloalkyloxy groups.

[0018] In some embodiments, R4 is selected from: methoxy, ethoxy, , , , .

[0019] In some implementations, R4 is... .

[0020] In some embodiments, R5 is selected from: methyl, trifluoromethyl, .

[0021] In some embodiments, R5 is a methyl group.

[0022] In some embodiments, the ring heteroatom in the R1-substituted or unsubstituted 5-7 membered heterocyclic group is one or two nitrogen atoms.

[0023] In some embodiments, the heteroatom on the ring of the 5-6-membered heteroaryl or C1-C3 alkyl-substituted 5-6-membered heteroaryl is one or two nitrogen atoms.

[0024] In some implementations, R is selected from: , .

[0025] In some implementations, R is selected from: .

[0026] In some implementations, R is .

[0027] In some implementations, R1 is selected from: C1-C6 linear alkyl groups, deuterated C1-C6 linear alkyl groups.

[0028] In some implementations, R1 is selected from: C1-C4 linear alkyl groups and deuterated C1-C3 linear alkyl groups.

[0029] In some implementations, R1 is selected from: Methyl, ethyl, n-butyl, deuterated methyl, deuterated ethyl.

[0030] In some embodiments, R3 is selected from: hydrogen, C1-C3 alkyl, fluorine-substituted C1-C3 alkyl, bromine-substituted C1-C3 alkyl, 3-membered heterocyclic group, 9-membered nitrogen-containing aryl group, etc. , , , , , ;

[0031] R6 and R7 are independently selected from: hydrogen, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, cyano, C3-C4 cycloalkyl, R9-substituted benzoyl, and R9-substituted naphthoyl.

[0032] R9 is selected from: hydrogen, C1-C3 alkoxy, C1-C3 alkyl;

[0033] R8 is selected from: hydrogen, C1-C3 alkyl.

[0034] In some embodiments, R9 is selected from hydrogen, methoxy, and ethoxy.

[0035] In some embodiments, R8 is selected from hydrogen, methyl, and ethyl.

[0036] In some embodiments, R3 is selected from: hydrogen, methyl, ethyl, difluoromethyl, difluoroethyl, ... , , , , , , , , , , , .

[0037] In some embodiments, R3 is selected from: hydrogen, methyl, ethyl, difluoromethyl, difluoroethyl, ... , , , , .

[0038] In some embodiments, R1 is selected from: difluoroacetyl, methyl, ethyl, n-butyl.

[0039] In some embodiments, R2 is selected from: methyl, ethyl, fluorine, chlorine.

[0040] In some embodiments, R2 is fluorine.

[0041] In a second aspect, the present invention provides the following applications of the pyrimidine diamine compounds or their pharmaceutically acceptable salts or stereoisomers.

[0042] The use of the pyrimidine diamine compounds or their pharmaceutically acceptable salts or stereoisomers described in this invention in the preparation of ALK inhibitors and / or FAK inhibitors.

[0043] The application of the pyrimidine diamine compounds of the present invention, or their pharmaceutically acceptable salts or stereoisomers, in the preparation of ALK and / or FAK protein degrading agents.

[0044] In some embodiments, the ALK is one or more of wild-type ALK, fusion mutant ALK, and point mutant ALK.

[0045] In some embodiments, the fusion mutant ALK is selected from one or more of EML4-ALK fusion protein, KIF5B-ALK fusion protein, TFG-ALK fusion protein, and CD74-ALK fusion protein.

[0046] In some embodiments, the point mutation type ALK is selected from one or more of L1196M mutation ALK, G1269A mutation ALK, G1202R mutation ALK, F1174C / L mutation ALK and I1171N mutation ALK.

[0047] The application of the pyrimidine diamine compound or its pharmaceutically acceptable salt or stereoisomer as described in this invention in the preparation of ALK inhibitors, wherein the ALK inhibitors are capable of inhibiting the protein activity of EML4-ALK fusion protein, EML4-ALK fusion protein L1196M drug-resistant mutant and / or EML4-ALK fusion protein G1269A drug-resistant mutant.

[0048] The present invention relates to the application of pyrimidine diamine compounds or their pharmaceutically acceptable salts or stereoisomers in the preparation of ALK degrading agents, wherein the ALK degrading agents are capable of degrading EML4-ALK fusion protein, EML4-ALK fusion protein L1196M drug-resistant mutant and / or EML4-ALK fusion protein G1269A drug-resistant mutant.

[0049] The use of the pyrimidine diamine compounds or their pharmaceutically acceptable salts or stereoisomers described in this invention in medicaments for the treatment and / or prevention of tumors.

[0050] In some embodiments, the tumor is a tumor caused by high ALK expression or abnormal ALK activation.

[0051] In some embodiments, the form of ALK aberrant activation is selected from one or more of fusion, point mutation, and amplification.

[0052] In some embodiments, the tumor is a tumor caused by ALK fusion mutations and / or ALK point mutations.

[0053] In some embodiments, the ALK fusion is selected from one or more of EML4-ALK fusion, KIF5B-ALK fusion, TFG-ALK fusion, and CD74-ALK fusion.

[0054] In some embodiments, the ALK point mutation is selected from one or more of the L1196M mutation, G1269A mutation, G1202R mutation, F1174C / L mutation, and I1171N mutation.

[0055] In some embodiments, the tumor is a tumor expressing the EML4-ALK fusion protein.

[0056] In some embodiments, the tumor is a tumor expressing a drug-resistant mutant of the EML4-ALK fusion protein L1196M.

[0057] In some embodiments, the tumor is a tumor expressing a drug-resistant mutant of the EML4-ALK fusion protein G1269A.

[0058] In some embodiments, the tumor is lung cancer, preferably non-small cell lung cancer.

[0059] Thirdly, the present invention provides a pharmaceutical composition prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises the pyrimidine diamine compound described in the present invention or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0060] The pyrimidine diamine compound provided by this invention is a molecular glue degrader targeting kinases such as ALK and FAK. It can effectively degrade wild-type ALK and FAK kinases and mutant ALK kinases, and can effectively inhibit the kinase activity of ALK and FAK and their mutants. It has a good anti-proliferative effect on tumors caused by high ALK expression or abnormal ALK activation. Furthermore, it has good selectivity, low toxicity, and good safety.

[0061] The pyrimidine diamine compounds of the present invention can induce selective ubiquitination and degradation of ALK and FAK proteins by utilizing the protein degradation system naturally present in human or animal cells, thereby transiently eliminating the biological functions of ALK and FAK proteins and reducing the activity of ALK and FAK proteins from the source. Compared with traditional inhibitors, they can significantly reduce the incidence of drug resistance mutations and effectively overcome drug resistance caused by existing drugs.

[0062] The compounds of this invention have excellent pharmacokinetic properties, good oral absorption efficiency and bioavailability, good drug-likeness, and their in vivo antitumor effects are significantly better than existing drugs such as ceritinib, showing great potential for clinical application. Attached Figure Description

[0063] Figure 1 The study aimed to assess the degradation activity of the compound against ALK and FAK in non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells.

[0064] Figure 2 The compound was used to assess its degradation activity against ALK in non-small cell lung cancer H3122 cells.

[0065] Figure 3 The compound was used to assess its degradation activity against ALK and FAK in H3122 non-small cell lung cancer cells.

[0066] Figure 4 The degradation effects of compound ZJL-4155 on ALK and FAK proteins in H3122 and Ba / F3-EML4-ALK cells were investigated at different concentrations and incubation times.

[0067] Figure 5 The levels of ALK and FAK proteins in H3122 and Ba / F3-EML4-ALK cells after treatment and elution with compound ZJL-4155.

[0068] Figure 6 The effect of compound ZJL4155 on the downstream signaling pathway of ALK.

[0069] Figure 7 To validate the degradation effect of compound ZJL-4155 on ALK and FAK proteins using whole-cell proteomics.

[0070] Figure 8 The effects of MG132 and Baf A1 on the degradation of ALK and FAK proteins mediated by compound ZJL-4155.

[0071] Figure 9 Iso TOP-ABPP validated that compound ZJL-4155 can bind to the Cys232 site of TRIM28.

[0072] Figure 10 The effect of TRIM28 gene knockout on the expression of ALK and FAK proteins in H3122 cells.

[0073] Figure 11 The degradation effects of compound W-2082 on ALK and FAK proteins in H3122 and Ba / F3-EML4-ALK cells at different concentrations and incubation times.

[0074] Figure 12 The levels of ALK and FAK proteins in H3122 and Ba / F3-EML4-ALK cells after treatment and elution with compound W-2082.

[0075] Figure 13 The effect of compound W-2082 on the downstream signaling pathway of ALK.

[0076] Figure 14 The effects of MG132, Baf A1, and Chloroquine on the degradation of ALK and FAK proteins mediated by compound W-2082.

[0077] Figure 15 Compound W-2082 is a p-type of BaF3-EML4-ALK G1269A ALK G1269A Degradation activity of mutants.

[0078] Figure 16 Compound W-2082 is a p-type of Ba / F3-EML4-ALK. G1269A The effects of the cell cycle.

[0079] Figure 17 Compound W-2082 is a p-type of Ba / F3-EML4-ALK. G1269A The effect of apoptosis.

[0080] Figure 18 The pharmacokinetic parameters of compound W-2082 are given.

[0081] Figure 19 This study describes the inhibitory effect of compound W-2082 on Ba / F3-EML4-ALK xenografts in vivo.

[0082] Figure 20 To investigate the in vivo activity of compound W-2082 against Ba / F3-EML4-ALK L1196M Inhibitory effect on transplanted tumors.

[0083] Figure 21 To investigate the in vivo activity of compound W-2082 against Ba / F3-EML4-ALK G1269A Inhibitory effect on transplanted tumors. Detailed Implementation

[0084] 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.

[0085] Unless otherwise specified, experimental methods in the following examples were performed under standard conditions, such as those described in the fourth edition of *Molecular Cloning: A Laboratory Manual*, edited by Green and Sambrook, published in 2013, or according to the manufacturer's recommendations. All commonly used chemical reagents used in the examples are commercially available products.

[0086] Unless otherwise defined, all technical and scientific terms used in this invention are consistent with those belonging to this invention.

[0087] The meanings are generally understood to be the same by those skilled in the art. 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.

[0088] 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."

[0089] 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 itself is 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.

[0090] 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.

[0091] 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.

[0092] 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.

[0093] 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), 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, dihydrothiopheneyl, dihydrotriazolyl, dihydroazacyclobutane, tetrahydrofuranyl, tetrahydrothiopheneyl, etc. , , , , , And so on, and their N-oxides. The connection of heterocyclic substituents can be achieved through carbon atoms or through heteroatoms.

[0094] 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.

[0095] This invention includes the free form of compounds of Formula I, 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 here not only include exemplary salts of the specific compounds described herein, but also typical pharmaceutically acceptable salts of the free forms of all Formula I compounds. The free form of a specific salt of the compound 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 a base, 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 the invention, such acid salts and base salts are otherwise pharmaceutically equivalent to their respective free forms.

[0096] 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.

[0097] 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.

[0098] 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.

[0099] 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.

[0100] 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 pyrimidine diamine compound or its pharmaceutically acceptable salt or stereoisomer as described in this invention) within a safe and effective range, and 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.

[0101] The "active ingredient" mentioned in this invention refers to the compound of formula I described in this invention, or its pharmaceutically acceptable salt or its stereoisomer.

[0102] The "active ingredient" and pharmaceutical composition described in this invention can be used to prepare drugs for the prevention and / or treatment of tumors.

[0103] "Safe and effective dose" means that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects.

[0104] "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.

[0105] "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.

[0106] Examples of pharmaceutically acceptable 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.), emulsifiers (such as Tween®), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0107] In another preferred embodiment, the compound of formula I of the present invention can form a complex with a macromolecular compound or polymer through non-bonding interaction. In another preferred embodiment, the compound of formula I of the present invention, as a small molecule, can also be linked to a macromolecular compound or polymer through chemical bonds. The macromolecular compound can be a biological macromolecule such as a polysaccharide, protein, nucleic acid, polypeptide, etc.

[0108] 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.

[0109] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0110] 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:

[0111] (a) Fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol and silica;

[0112] (b) Adhesives, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0113] (c) Moisturizers, such as glycerin;

[0114] (d) Disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginate, certain complex silicates, and sodium carbonate;

[0115] (e) Slow solvents, such as paraffin;

[0116] (f) Absorption accelerators, for example, quaternary ammonium compounds;

[0117] (g) Wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0118] (h) Adsorbents, such as kaolin; and

[0119] (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.

[0120] 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.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] The present invention will be further described in detail below with reference to specific embodiments.

[0125] Example 1: Synthesis of compounds ZJL-4132 to ZJL-5056

[0126]

[0127] 2-Cyano-3-cyclopropylacrylic acid (28.0 mg, 0.20 mmol), HATU (97.0 mg, 0.26 mmol), DIEA (59.2 μL, 0.34 mmol), and N,N-dimethylformamide (3 ml) were added to a 25 ml round-bottom flask and stirred for 10 min. Ceritinib (100 mg, 0.17 mmol) was added, and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After the reaction was completed, water was added to quench the reaction. The mixture was extracted with 10 ml of ethyl acetate, washed three times with 20 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation and purified by silica gel column chromatography (DCM:MeOH = 20:1-30:1) to give a pale white solid (60 mg, 52.2%). 1 H NMR (400 MHz, Chloroform-d) δ 9.49 (s, 1H), 8.56 (d, J= 8.4 Hz, 1H), 8.14 (s, 1H), 8.02 (s, 1H), 7.92 (dd, J = 7.9, 1.6 Hz, 1H), 7.62 (ddd, J = 8.6, 7.4, 1.7 Hz, 1H), 7.55 (s, 1H), 7.30 – 7.21 (m, 1H), 6.70(s, 1H), 6.60 (d, J = 11.2 Hz, 1H), 4.55 (p, J = 6.1 Hz, 1H), 3.25 (p, J =6.9 Hz, 1H), 2.93 (ddd, J = 12.1, 8.1, 3.4 Hz, 2H), 2.17 (s, 3H), 2.08 (qd, J= 7.7, 3.7 Hz, 1H), 1.95 – 1.77 (m, 3H), 1.67 (q, J = 13.5, 12.0 Hz, 2H), 1.35 (d, J = 6.0 Hz, 6H), 1.31 (d, J = 6.9 Hz, 6H), 1.27 – 1.18 (m, 4H), 0.89 (dd, J = 4.4, 2.6 Hz, 1H). 13C NMR (151 MHz, Chloroform-d) δ 165.57, 162.31,157.50, 155.42, 155.39, 144.80, 138.54, 136.28, 134.71, 131.35, 128.09,127.00, 124.98, 123.73, 123.22, 120.79, 115.71, 110.95, 107.91, 71.81, 55.53,38.29, 29.77, 22.32, 19.08, 15.64, 15.44, 10.68. HRMS (ESI) calcd.forC 35 H 41 ClN6O4S 677.2671 [M + H] + , found 677.2645.

[0128]

[0129] The synthesis method of ZJL-4147 is similar to that of ZJL-4132, yielding a yellow solid (45 mg, 37.8%). 1 H NMR (400MHz, Chloroform-d) δ 9.55 (s, 1H), 8.58 (d, J = 8.4 Hz, 1H), 8.15 (s, 1H), 8.02 (s, 1H), 7.92 (d, J = 6.8 Hz, 3H), 7.72 – 7.59 (m, 3H), 7.56 (t, J = 7.3Hz, 2H), 7.28 (d, J = 7.7 Hz, 1H), 6.73 (d, J = 5.0 Hz, 1H), 4.81 (d, J =13.3 Hz, 1H), 4.69 – 4.49 (m, 2H), 3.46 – 3.35 (m, 1H), 3.26 (p, J = 6.8 Hz,1H), 3.14 (s, 1H), 2.99 (dd, J = 13.7, 10.2 Hz, 1H), 2.76 (t, J = 12.3 Hz,1H), 2.21 (s, 3H), 1.96 (d, J = 15.5 Hz, 1H), 1.82 (s, 2H), 1.67 (d, J = 12.6Hz, 2H), 1.38 (t, J = 5.7 Hz, 6H), 1.32 (d, J = 6.8 Hz, 6H). 13C NMR (151 MHz, Chloroform-d) δ 157.53, 155.44, 144.90, 138.59, 136.63, 134.75, 133.62,131.40, 130.47, 129.31, 126.79, 125.03, 123.78, 123.25, 120.77, 110.83,105.97, 71.67, 46.73, 43.66, 38.52, 33.99, 33.72, 32.39, 32.14, 29.10, 22.41,19.13, 15.49. HRMS (ESI) calcd.for C 37 H 41 ClN6O4S 701.2671 [M + H] + , found701.2642.

[0130]

[0131] The synthesis method of ZJL-4148 is similar to that of ZJL-4132, yielding a yellow solid (53 mg, 42.6%). 1 H NMR (400MHz, Chloroform-d) δ 9.52 (s, 1H), 8.58 (dd, J = 8.4, 1.1 Hz, 1H), 8.16 (s,1H), 8.04 (s, 1H), 8.01 (d, J = 13.7 Hz, 1H), 7.93 (dd, J = 8.0, 1.6 Hz, 1H),7.65 – 7.58 (m, 4H), 7.58 – 7.54 (m, 2H), 7.53 (s, 1H), 6.72 (s, 1H), 4.91(s, 1H), 4.57 (p, J = 6.1 Hz, 1H), 3.84 (s, 1H), 3.25 (dq, J = 17.0, 10.1,8.5 Hz, 2H), 3.01 – 2.84 (m, 2H), 2.18 (s, 3H), 1.77 (t, J = 55.5 Hz, 4H), 1.38 (d, J = 6.0 Hz, 6H), 1.32 (d, J = 6.9 Hz, 6H). 13C NMR (151 MHz, Chloroform-d) δ 157.27, 155.25, 144.63, 139.49, 138.37, 137.86, 137.80,136.11, 134.51, 131.22, 131.14, 129.14, 127.96, 126.91, 124.83, 123.50,123.05, 120.67, 110.76, 105.87, 71.71, 55.37, 38.24, 18.90, 15.28. HRMS (ESI)calcd.for C 37 H 41 ClN6O6S 733.2570 [M + H] + , found 733.2538.

[0132]

[0133] The synthesis method of ZJL-4149 is similar to that of ZJL-4132, yielding a white solid (56 mg, 44.2%). 1 H NMR (400MHz, Chloroform-d) δ 9.51 (s, 1H), 8.57 (dd, J = 8.4, 1.0 Hz, 1H), 8.15 (s,1H), 8.09 – 8.03 (m, 3H), 8.02 – 7.90 (m, 2H), 7.62 (ddd, J = 8.6, 7.3, 1.7Hz, 1H), 7.59 – 7.52 (m, 2H), 6.98 (d, J = 8.9 Hz, 2H), 6.68 (s, 1H), 4.91(d, J = 13.2 Hz, 1H), 4.54 (p, J = 6.1 Hz, 1H), 4.24 (d, J = 13.6 Hz, 1H),3.90 (s, 3H), 3.32 – 3.20 (m, 2H), 3.03 – 2.92 (m, 1H), 2.80 (td, J = 13.1,2.7 Hz, 1H), 2.19 (s, 3H), 1.89 (d, J = 13.3 Hz, 2H), 1.64 (ddd, J = 22.6,12.9, 3.9 Hz, 3H), 1.36 (d, J = 6.1 Hz, 6H), 1.32 (d, J = 6.9 Hz, 7H). 13C NMR(151 MHz, Chloroform-d) δ 164.29, 164.14, 155.46, 144.86, 138.58, 136.35,134.75, 134.38, 132.14, 131.43, 131.41, 130.12, 128.05, 126.95, 125.06,123.79, 123.28, 120.84, 114.23, 110.80, 106.00, 71.73, 55.71, 55.58, 47.17,43.48, 38.40, 33.70, 32.47, 22.37, 22.34, 19.12, 15.49. HRMS (ESI) calcd.forC 39 H 44 ClN5O6S 746.2774 [M + H] + , found 746.2742.

[0134]

[0135] The synthesis method of ZJL-4151 is similar to that of ZJL-4132, yielding a white solid (42 mg, 39.6%). 1H NMR (400MHz, Chloroform-d) δ 9.50 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.15 (s, 1H), 8.02 (s, 1H), 7.92 (dd, J = 7.9, 1.6 Hz, 1H), 7.65 – 7.59 (m, 1H), 7.55 (s,1H), 6.67 (s, 1H), 4.74 (d, J = 13.1 Hz, 1H), 4.62 – 4.49 (m, 2H), 3.26 (p, J= 6.8 Hz, 1H), 3.17 (td, J = 13.0, 2.6 Hz, 1H), 2.91 (tt, J = 12.2, 3.4 Hz,1H), 2.72 (td, J = 13.0, 2.8 Hz, 1H), 2.18 (s, 3H), 2.02 (s, 3H), 1.83 (ddt,J = 18.7, 13.1, 2.7 Hz, 2H), 1.58 (ddd, J = 17.0, 12.5, 4.2 Hz, 2H), 1.36(dd, J = 6.1, 3.5 Hz, 6H), 1.31 (d, J = 6.8 Hz, 6H). 13 C NMR (151 MHz, Chloroform-d) δ 157.29, 155.20, 144.60, 138.35, 136.31, 134.51, 131.17,127.75, 126.68, 124.78, 123.52, 123.02, 120.57, 110.58, 105.75, 89.28, 71.48,55.34, 47.81, 41.96, 38.15, 33.12, 32.09, 22.15, 22.10, 18.88, 15.25, 4.02. (ESI) calcd.for C 32 H 38 ClN5O4S 624.24O6 [M + H] + , found 624.2374.

[0136]

[0137] The first step, the synthesis method of ZJL-4152-R is similar to that of ZJL-4132.

[0138] In the second step, ZJL-4152-R (140.0 mg, 0.16 mmol), TFA (98.8 mg, 1.29 mmol), TES (103.0 mg, 0.64 mmol), DIEA (280.8 μL, 1.16 mmol), and DCM (3 ml) were added to a 5 ml round-bottom flask under 0 °C ice bath conditions. The reaction was carried out for 1 h, and the reaction was monitored by TLC. After the reaction was completed, water was added to quench the reaction, and the product was extracted with 10 ml of ethyl acetate. The organic phase was washed three times with 20 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (DCM:MeOH = 20:1-30:1) to obtain a white solid (30 mg, 28.2%). 1 H NMR (400 MHz, Chloroform-d) δ 9.50 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.14 (s, 1H), 8.03 (s, 1H), 7.92(dd, J = 8.0, 1.7 Hz, 1H), 7.65 – 7.58 (m, 1H), 7.55 (d, J = 3.7 Hz, 1H), 7.24 (d, J = 7.5 Hz, 1H), 6.68 (d, J = 15.9 Hz, 1H), 4.76 (t, J = 14.1 Hz,1H), 4.54 (dp, J = 17.9, 6.1 Hz, 1H), 4.29 (d, J = 13.5 Hz, 1H), 3.35 – 3.15(m, 2H), 2.95 (qt, J = 12.0, 3.3 Hz, 1H), 2.76 (ddd, J = 13.6, 10.5, 7.1 Hz,2H), 2.18 (s, 3H), 2.05 – 1.76 (m, 5H), 1.61 (dtd, J = 20.1, 11.7, 10.9, 3.8Hz, 2H), 1.38 – 1.34 (m, 6H), 1.30 (d, J = 6.8 Hz, 7H). 13C NMR (151 MHz, Chloroform-d) δ 157.50, 155.43, 155.39, 144.82, 138.55, 134.70, 131.36,128.02, 126.85, 124.98, 123.72, 123.69, 123.21, 120.82, 120.77, 110.86,110.77, 105.97, 71.78, 71.69, 55.53, 45.96, 45.76, 43.88, 43.67, 38.45,38.30, 33.69, 33.19, 32.53, 32.09, 29.77, 22.35, 22.30, 19.08, 15.45. HRMS(ESI) calcd.for C 31 H 39 ClN6O4S 627.2515 [M + H] + , found 627.2493.

[0139]

[0140] The synthesis method of ZJL-4153 is similar to that of ZJL-4132, yielding a white solid (55 mg, 51.6%). 1H NMR (400MHz, Chloroform-d) δ 9.51 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.15 (s, 1H), 8.04 (s, 1H), 7.93 (dd, J = 7.9, 1.7 Hz, 1H), 7.66 – 7.58 (m, 1H), 7.55 (s,1H), 7.27 (d, J = 6.9 Hz, 1H), 6.68 (d, J = 6.9 Hz, 1H), 4.76 (dd, J = 13.0,6.7 Hz, 1H), 4.61 – 4.48 (m, 1H), 4.31 (d, J = 13.6 Hz, 1H), 3.68 (dd, J =4.4, 2.3 Hz, 1H), 3.25 (dd, J = 13.6, 6.8 Hz, 2H), 3.08 – 2.86 (m, 3H), 2.79– 2.67 (m, 1H), 2.18 (s, 3H), 1.93 – 1.78 (m, 2H), 1.72 – 1.52 (m, 2H), 1.37(s, 6H), 1.31 (d, J = 6.9 Hz, 6H). 13 C NMR (151 MHz, Chloroform-d) δ 157.52,155.46, 155.43, 144.84, 138.58, 136.35, 134.73, 131.39, 128.06, 126.89,125.01, 123.74, 123.24, 120.83, 120.78, 110.83, 106.00, 71.79, 71.74, 55.56,48.29, 48.21, 43.40, 43.24, 30.24, 29.80, 22.38, 19.10, 15.47, 14.23. HRMS(ESI) calcd.for C 31 H 38 ClN5O5S 628.2355 [M + H] + , found 628.2343.

[0141]

[0142] The synthesis method of ZJL-4154 is similar to that of ZJL-4132, yielding a white solid (52 mg, 50.0%). 1H NMR (400MHz, Chloroform-d) δ 9.50 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.15 (s, 1H),8.02 (s, 1H), 7.92 (dd, J = 8.0, 1.6 Hz, 1H), 7.66 – 7.57 (m, 1H), 7.55 (s,1H), 7.25 (s, 1H), 6.70 – 6.58 (m, 2H), 6.32 (dd, J = 16.8, 2.0 Hz, 1H), 5.70(dd, J = 10.5, 2.0 Hz, 1H), 4.86 (d, J = 13.5 Hz, 1H), 4.53 (p, J = 6.1 Hz,1H), 4.20 – 4.05 (m, 1H), 3.30 – 3.11 (m, 2H), 2.93 (tt, J = 12.0, 3.5 Hz,1H), 2.73 (t, J = 13.4 Hz, 1H), 2.18 (s, 3H), 1.84 (d, J = 13.2 Hz, 2H), 1.71– 1.49 (m, 2H), 1.35 (d, J = 6.2 Hz, 7H), 1.31 (d, J = 6.9 Hz, 6H). 13 C NMR(151 MHz, Chloroform-d) δ 165.51, 157.55, 155.47, 155.45, 144.84, 138.60,136.61, 134.75, 131.41, 127.99, 127.93, 127.82, 126.95, 125.04, 123.77,123.25, 120.81, 110.86, 105.99, 71.72, 55.57, 46.90, 43.14, 38.47, 29.82,22.39, 22.33, 19.12, 15.49. HRMS (ESI) calcd.for C 31 H 38 ClN5O4S 612.2406 [M + H] + , found 612.2385.

[0143]

[0144] The synthesis method of ZJL-4155 is similar to that of ZJL-4132, yielding a white solid (48 mg, 44.5%). 1 H NMR (400MHz, Chloroform-d) δ 9.51 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.16 (s, 1H), 8.04 (s, 1H), 7.93 (dd, J = 8.0, 1.6 Hz, 1H), 7.66 – 7.59 (m, 1H), 7.55 (s,1H), 7.27 (d, J = 7.9 Hz, 1H), 6.68 (s, 1H), 6.16 (t, J = 53.8 Hz, 1H), 4.76– 4.64 (m, 1H), 4.54 (q, J = 6.1 Hz, 1H), 4.24 (d, J = 13.7 Hz, 1H), 3.32 –3.17 (m, 2H), 2.96 (ddt, J = 11.9, 7.3, 3.5 Hz, 1H), 2.81 (t, J = 12.8 Hz,1H), 2.18 (s, 3H), 1.93 – 1.83 (m, 2H), 1.64 (dt, J = 13.0, 3.9 Hz, 2H), 1.37(dd, J = 6.0, 2.6 Hz, 6H), 1.32 (d, J = 6.9 Hz, 6H). 13 C NMR (151 MHz, Chloroform-d) δ 157.31, 155.25, 155.23, 144.65, 138.38, 135.88, 134.54,131.21, 127.98, 126.78, 124.84, 123.55, HRMS (ESI) calcd.for C 30 H 36 ClF2N5O4S 636.2217 [M + H] + , found 636.2195.

[0145]

[0146] The synthesis method of ZJL-5003 is similar to that of ZJL-4132, yielding a white solid (28 mg, 25.4%). 1 H NMR (400MHz, Chloroform-d) δ 9.51 (s, 1H), 8.57 (d, J = 8.3 Hz, 1H), 8.16 (s, 1H), 8.03 (s, 1H), 7.93 (dd, J = 7.9, 1.6 Hz, 1H), 7.62 (ddd, J = 8.7, 7.4, 1.7Hz, 1H), 7.55 (s, 1H), 7.24 (d, J = 7.1 Hz, 1H), 6.73 (s, 1H), 6.48 (dd, J =16.6, 9.9 Hz, 1H), 6.27 (d, J = 16.7 Hz, 1H), 6.06 (d, J = 9.9 Hz, 1H), 4.60– 4.50 (m, 1H), 3.90 (dt, J = 11.3, 2.3 Hz, 2H), 3.26 (hept, J = 6.8 Hz, 1H), 2.72 (tdd, J = 11.7, 8.6, 3.4 Hz, 3H), 2.15 (s, 3H), 1.91 – 1.73 (m, 4H), 1.38 (d, J = 6.0 Hz, 6H), 1.32 (d, J = 6.8 Hz, 7H). 13 C NMR (151 MHz, Chloroform-d) δ 157.51, 155.47, 144.89, 138.61, 136.35, 134.73, 132.73,131.42, 128.62, 128.06, 126.89, 125.07, 123.77, 123.26, 120.79, 110.81,106.05, 71.77, 55.59, 46.77, 37.61, 32.37, 22.37, 19.06, 15.50. HRMS (ESI)calcd.for C 30 H 38 ClN5O5S2 648.2076 [M + H] + , found 648.2048.

[0147]

[0148] The synthesis method of ZJL-5032 is similar to that of ZJL-4132, yielding a white solid (46 mg, 34.2%). 1 H NMR (400MHz, Chloroform-d) δ 9.50 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.15 (s, 1H), 8.03 (s, 1H), 7.93 (dd, J = 8.0, 1.6 Hz, 1H), 7.62 (ddd, J = 8.7, 7.3, 1.7Hz, 1H), 7.55 (s, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.26 – 7.23 (m, 1H), 7.13(s, 1H), 7.10 (s, 1H), 6.72 (s, 1H), 6.62 (d, J = 7.4 Hz, 1H), 4.57 (p, J =6.1 Hz, 1H), 4.34 – 4.14 (m, 2H), 3.26 (p, J = 6.9 Hz, 1H), 2.92 (d, J = 12.9Hz, 2H), 2.18 (s, 3H), 1.77 (d, J = 27.1 Hz, 6H), 1.41 (d, J = 7.1 Hz, 3H), 1.38 (d, J = 6.1 Hz, 6H), 1.31 (d, J = 6.9 Hz, 6H). 13 C NMR (151 MHz, Chloroform-d) δ 170.18, 157.44, 155.36, 155.34, 144.73, 140.43, 138.48,136.54, 134.64, 131.30, 128.77, 127.96, 126.95, 124.91, 123.65, 123.15,120.74, 117.65, 117.60, 110.88, 105.90, 71.74, 62.54, 62.51, 55.47, 38.45,22.28, 19.01, 16.18, 16.13, 15.38. HRMS (ESI) calcd.for C 39 H 46 ClN6O6PS 793.2698[M + H] + , found 793.2683.

[0149]

[0150] The synthesis method of ZJL-5056 is similar to that of ZJL-4132, yielding a white solid (47 mg, 67.8%). 1 H NMR (400MHz, CDCl3) δ 9.47 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.13 (s, 1H), 7.98 (s,1H), 7.91 (dd, J = 8.0, 1.6 Hz, 1H), 7.64 – 7.57 (m, 1H), 7.54 (s, 1H), 7.24(s, 1H), 6.82 (s, 1H), 4.53 (hept, J = 6.1 Hz, 1H), 3.25 (p, J = 6.9 Hz, 1H), 3.16 (dd, J = 8.8, 5.8 Hz, 2H), 2.69 (tt, J = 11.9, 3.8 Hz, 1H), 2.49 – 2.41(m, 2H), 2.14 (s, 5H), 1.87 (qd, J = 12.4, 3.5 Hz, 2H), 1.80 – 1.72 (m, 2H),1.62 – 1.52 (m, 2H), 1.32 (dd, J = 16.1, 6.5 Hz, 14H), 0.93 (t, J = 7.3 Hz, 3H). HRMS (ESI) calcd.for C 37 H 41 ClN6O4S 701.2671 [M + H] + , found 701.2637.

[0151] Example 2: Synthesis of compounds W-2082 to W-2083

[0152]

[0153] Ceritinib (60.0 mg, 0.11 mmol), CH3I (17 mg, 0.12 mmol), K2CO3 (23 mg, 0.16 mmol), and DMF (3 ml) were added to a 25 ml round-bottom flask and reacted at room temperature for 4 h. The reaction was monitored by TLC. After the reaction was completed, water was added to quench the reaction, and the product was extracted with 20 ml of ethyl acetate. The organic phase was washed three times with 20 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (DCM:MeOH = 20:1-30:1) to give a white solid (32 mg, 52.3%). 1H NMR (400 MHz, CDCl3) δ 9.48 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.14 (s, 1H), 7.99 (s, 1H), 7.92 (d, J = 7.9 Hz, 1H), 7.66 – 7.49 (m, 2H),7.24 (d, J = 7.5 Hz, 1H), 6.81 (s, 1H), 4.52 (p, J = 6.1 Hz, 1H), 3.25 (h, J= 6.9 Hz, 1H), 3.05 (d, J = 11.3 Hz, 2H), 2.70 – 2.63 (m, 1H), 2.38 (s, 3H), 2.15 (s, 5H), 1.88 – 1.73 (m, 4H), 1.35 (d, J = 6.0 Hz, 6H), 1.31 (d, J = 6.9Hz, 6H). 13 C NMR (151 MHz, CDCl3) δ 155.33, 155.31, 138.48, 134.64, 131.26,127.88, 126.67, 124.90, 123.68, 123.13, 120.57, 114.07, 110.86, 105.84,77.26, 77.05, 76.83, 55.71, 55.45, 44.92, 36.44, 31.93, 31.51, 31.44, 31.15,30.31, 30.19, 30.14, 29.78, 29.70, 29.66, 29.37, 22.70, 22.22, 18.97, 15.37,14.13. HRMS (ESI) calcd.for C 29 H 38 ClN5O3S 572.2457 [M + H] + , found 572.2423.

[0154]

[0155] The synthesis method of ZJL-5055 is similar to that of W-2082, yielding a white solid (57 mg, 43.3%). 1H NMR (400MHz, CDCl3) δ 9.47 (s, 1H), 8.57 (d, J = 8.4 Hz, 1H), 8.13 (s, 1H), 7.98 (s,1H), 7.91 (dd, J = 8.0, 1.6 Hz, 1H), 7.64 – 7.58 (m, 1H), 7.54 (s, 1H), 7.24(s, 1H), 6.82 (s, 1H), 4.54 (h, J = 6.0 Hz, 1H), 3.25 (p, J = 6.9 Hz, 1H), 3.16 (dd, J = 8.8, 5.8 Hz, 2H), 2.69 (tt, J = 11.9, 3.9 Hz, 1H), 2.49 – 2.41 (m, 2H), 2.14 (s, 5H), 1.87 (qd, J = 12.4, 3.5 Hz, 2H), 1.81 – 1.73 (m, 2H), 1.61 – 1.52 (m, 2H), 1.34 (d, J = 6.2 Hz, 6H), 1.30 (d, J = 6.9 Hz, 6H), 0.93(t, J = 7.3 Hz, 3H). HRMS (ESI) calcd.for C 32 H 44 ClN5O3S 614.2926 [M + H] + ,found 614.2899.

[0156]

[0157] The synthesis method of W-2083 is similar to that of W-2082, yielding a yellow solid (50 mg, 79.8%). 1H NMR (400 MHz, CDCl3) δ 9.49 (s, 1H), 8.57 (d, J = 8.3 Hz, 1H), 8.15 (s, 1H), 8.01 (s, 1H), 7.92 (dd, J = 8.1, 1.7 Hz, 1H), 7.65 – 7.53 (m, 2H), 7.24 (d, J = 7.6 Hz, 1H), 6.85 (s, 1H), 4.59 (p, J = 6.0 Hz, 1H), 3.91 (s, 2H), 3.41 (d, J = 11.4Hz, 2H), 3.26 (p, J = 6.8 Hz, 1H), 2.81 (q, J = 7.3 Hz, 3H), 2.43 (t, J =11.7 Hz, 2H), 2.15 (s, 5H), 2.02 (s, 1H), 1.85 (d, J = 13.6 Hz, 2H), 1.36 (d,J = 6.1 Hz, 6H), 1.32 – 1.30 (m, 6H). 13 C NMR (151 MHz, CDCl3) δ 157.55,155.43, 144.96, 138.59, 136.20, 134.75, 131.37, 127.95, 126.63, 125.01,123.80, 123.23, 120.62, 110.95, 105.94, 77.37, 77.16, 76.95, 71.67, 55.56,53.36, 52.39, 37.08, 32.04, 31.62, 31.05, 29.81, 29.78, 29.74, 22.35, 19.09,15.48. HRMS (ESI) calcd.for C 30 H 40 ClN5O3S 586.2613 [M + H] + , found 586.2592.

[0158] Example 3: Synthesis of compound W-3040

[0159]

[0160] Ceritinib (70.0 mg, 0.12 mmol), dibromoacetic acid (31 mg, 0.14 mmol), N,N,N',N'-tetramethylchloromethamidine hexafluorophosphate (TCFH, 40 mg, 0.14 mmol), N-methylimidazole (NMI, 21 mg, 0.26 mmol), and CH3CN (4 ml) were added to a 25 ml round-bottom flask and reacted overnight at room temperature. The reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated and the product was purified by silica gel column chromatography (DCM:MeOH = 100:1) to give a white solid (58 mg, 64.0%). 1 H NMR (400 MHz, DMSO) δ9.46 (s, 1H), 8.46 (d, J = 8.3 Hz, 1H), 8.25 (s, 1H), 8.06 (s, 1H), 7.87 –7.81 (m, 1H), 7.63 (t, J = 7.8 Hz, 1H), 7.53 (s, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.22 (s, 1H), 6.81 (s, 1H), 4.56 (p, J = 6.0 Hz, 1H), 4.48 (d, J = 12.8Hz, 1H), 4.14 (d, J = 13.5 Hz, 1H), 3.45 (q, J = 6.8 Hz, 1H), 3.21 (t, J =12.5 Hz, 1H), 2.96 (dd, J = 13.4, 10.0 Hz, 1H), 2.84 (t, J = 12.5 Hz, 1H), 2.15 (s, 3H), 1.78 – 1.70 (m, 2H), 1.65 – 1.54 (m, 2H), 1.19 (d, J = 6.0 Hz, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS (ESI) calcd.for C 30 H 36 Br₂ClN₅O₄S 756.0616[M+H] + , found 756.0598.

[0161] Example 4: Synthesis of compounds W-3071 and W-3074

[0162]

[0163] In the first step, 2-bromo-4-fluoro-5-nitrotoluene (500 mg, 2.14 mmol), cesium carbonate (1392 mg, 4.27 mmol), and anhydrous ethanol (6 ml) were added to a 25 ml round-bottom flask and reacted overnight at 60 °C. The reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated, and the product was extracted with 20 ml of ethyl acetate. The organic phase was washed three times with 30 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 40:1-60:1) to obtain a yellow solid (546 mg, 98.0%). 1H NMR (400 MHz, DMSO) δ 7.90 (s, 1H), 7.59 (s, 1H), 4.21 (q, J = 6.9 Hz, 2H), 2.32 (s, 3H), 1.31 (t, J = 7.0 Hz, 3H).

[0164] In the second step, 1-bromo-5-ethoxy-2-methyl-4-nitrobenzene (546 mg, 2.10 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (714 mg, 2.31 mmol), tetra(triphenylphosphine)palladium (242 mg, 0.21 mmol), cesium carbonate (1710 mg, 5.25 mmol), and a mixed solvent of 1,4-dioxane:water = 2:1 (12 ml) were added to a 25 ml round-bottom flask. The mixture was reacted overnight at 100 °C under argon protection. The reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated to dryness, and the product was extracted with 30 ml of ethyl acetate, filtered through diatomaceous earth, concentrated by vacuum distillation, and purified by silica gel column chromatography (PE:EA = 10:1-30:1) to obtain a white solid (252 mg, 65.0%). 1 H NMR (400 MHz, DMSO) δ 7.71 (s, 1H), 7.03 (s, 1H), 5.68 (s,1H), 4.18 (q, J = 6.9 Hz, 2H), 3.97 (d, J = 3.4 Hz, 2H), 3.54 (t, J = 5.6 Hz,2H), 2.31 (s, 2H), 2.21 (s, 3H), 1.43 (s, 9H), 1.31 (t, J = 6.9 Hz, 3H).

[0165] In the third step, 3071-S3 (670 mg, 1.85 mmol), 10% Pd / C (201 mg, 30 wt%), and methanol (6 ml) were added to a 25 ml round-bottom flask. The mixture was reacted overnight under a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was completed, the mixture was extracted with 30 ml of ethyl acetate, filtered through diatomaceous earth, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 10:1-20:1) to obtain a colorless oily liquid (579 mg, 93.7%). 1 H NMR (400 MHz, DMSO) δ 6.56 (s, 1H), 6.41 (s, 1H), 4.39 (s, 2H), 4.04 (dd, J = 13.5, 8.3 Hz, 2H), 3.94 (q, J = 6.9 Hz, 2H), 2.85– 2.63 (m, 3H), 2.11 (s, 3H), 1.59 (d, J = 12.9 Hz, 2H), 1.41 (s, 9H), 1.29(t, J = 6.9 Hz, 3H).

[0166] In the fourth step, 3071-S4 (300 mg, 0.90 mmol), 2,5-dichloro-N-[2-[(1-methylethyl)sulfonyl]phenyl]-4-pyrimidinamine (342 mg, 0.99 mmol), palladium acetate (10 mg, 0.045 mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (43 mg, 0.09 mmol), cesium carbonate (880 mg, 2.70 mmol), and tetrahydrofuran (6 ml) were added to a 10 ml sealed tube. The mixture was microwaved at 110 °C for 1 h under an argon atmosphere. The reaction was monitored by TLC. After the reaction was completed, the mixture was extracted with 30 ml of ethyl acetate, filtered through diatomaceous earth, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain a white solid (60 mg, 10%). 1H NMR (400 MHz, DMSO) δ 9.48 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 8.20 (d, J = 31.6 Hz, 2H), 7.83 (dd, J = 7.9, 1.6 Hz, 1H), 7.60 (t, J = 7.8 Hz,1H), 7.47 (s, 1H), 7.34 (t, J = 7.6 Hz, 1H), 6.82 (s, 1H), 4.16 – 3.99 (m,4H), 3.44 (p, J = 6.8 Hz, 1H), 2.82 (d, J = 10.6 Hz, 3H), 2.15 (s, 3H), 1.66(d, J = 12.6 Hz, 2H), 1.59 – 1.50 (m, 2H), 1.42 (s, 9H), 1.30 – 1.25 (m, 3H), 1.16 (d, J = 6.8 Hz, 6H).

[0167] In step 5, 3071-S5 (60 mg, 0.09 mmol), HCl in 1,4-dioxane solution (2 ml), and DCM (2 ml) were added to a 25 ml round-bottom flask. After reacting for 2 h, the mixture was concentrated by vacuum distillation. Then, CH3I (13 mg, 0.09 mmol), K2CO3 (17 mg, 0.12 mmol), and DMF (3 ml) were added. The mixture was reacted at room temperature for 4 h, and the reaction was monitored by TLC. After the reaction was completed, the mixture was quenched with water, extracted with 20 ml of ethyl acetate, washed three times with 20 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (DCM:MeOH = 10:1-20:1) to obtain a white solid (24 mg, 53.8%). 1H NMR(400 MHz, DMSO) δ 9.48 (s, 1H), 8.47 (d, J = 8.4 Hz, 1H), 8.20 (d, J = 24.1Hz, 2H), 7.82 (dd, J = 7.9, 1.6 Hz, 1H), 7.63 – 7.55 (m, 1H), 7.46 (s, 1H), 7.38 – 7.30 (m, 1H), 6.81 (s, 1H), 4.03 (q, J = 6.9 Hz, 2H), 2.99 (d, J =11.1 Hz, 2H), 2.69 – 2.61 (m, 1H), 2.31 (s, 3H), 2.24 – 2.15 (m, 2H), 2.13(s, 3H), 1.79 – 1.64 (m, 4H), 1.26 (t, J = 6.9 Hz, 3H), 1.16 (d, J = 6.8 Hz,6H). HRMS (ESI) calcd.for C 28 H 36 ClN5O3S 558.2300 [M+H] + Found 558.2285.

[0168]

[0169] The synthesis method of W-3074 is similar to that of W-3071, yielding a white solid (23 mg, total yield 1.9%). 1H NMR(400 MHz, CDCl3) δ 9.51 (s, 1H), 8.60 – 8.54 (m, 1H), 8.15 (s, 1H), 8.01 (s,1H), 7.92 (dd, J = 8.0, 1.7 Hz, 1H), 7.62 (ddd, J = 8.7, 7.3, 1.7 Hz, 1H), 7.51 (s, 1H), 7.24 (d, J = 8.3 Hz, 1H), 6.81 (s, 1H), 3.88 (s, 3H), 3.41 (d,J = 11.6 Hz, 2H), 3.25 (p, J = 6.9 Hz, 1H), 2.86 – 2.76 (m, 1H), HRMS (ESI) calcd.for C 27 H 34 ClN5O3S544.2144 [M+H] + , found 544.2126.

[0170] Example 5: Synthesis of compound W-3076

[0171]

[0172] The first step was similar to the synthesis method of ZJL-4132, yielding a white solid (431 mg, 75.2%). 1 H NMR(400 MHz, DMSO) δ 8.42 (q, J = 4.6 Hz, 1H), 7.91 (s, 1H), 7.71 (s, 1H), 4.97(m, 1H), 2.74 (d, J = 4.6 Hz, 3H), 1.28 (d, J = 6.0 Hz, 6H).

[0173] The second step involved a synthesis method similar to step 2 of the W-3071 synthesis, yielding a yellow solid (489 mg, 85.7%).

[0174] The third step involved a synthesis method similar to step 3 of the W-3071 synthesis, yielding a white solid (335 mg, 73.5%).

[0175] Fourth step: Following a similar synthesis method as step 4 of W-3071 synthesis, a white solid (120 mg, 33.5%) was obtained. 1 HNMR (400 MHz, CDCl3) δ 9.63 (d, J = 6.3 Hz, 1H), 8.56 (d, J = 8.4 Hz, 1H), 8.37 (m, 1H), 8.17 (s, 1H), 7.90 (m, 1H), 7.72 – 7.61 (m, 1H), 7.58 (s, 1H),7.23 (d, J = 7.6 Hz, 1H), 6.74 (d, J = 45.3 Hz, 1H), 5.75 (m, 1H), 4.66 (h, J= 6.1 Hz, 1H), 4.18 (m 1H), 4.06 (s, 1H), 3.58 (t, J = 5.5 Hz, 1H), 3.25 (m,1H), 2.80 (dd, J = 31.9, 4.8 Hz, 4H), 2.33 (s, 1H), 1.84 (d, J = 12.9 Hz,1H), 1.49 (d, J = 2.7 Hz, 9H), 1.40 (dd, J = 6.1, 2.4 Hz, 6H), 1.32 (dd, J =6.8, 2.6 Hz, 6H).

[0176] Fifth step: Following a similar synthesis method as step 5 of W-3071 synthesis, a white solid (40 mg, 38.3%) was obtained. 1HNMR (400 MHz, CDCl3) δ 9.64 (d, J = 6.3 Hz, 1H), 8.60 – 8.52 (m, 1H), 8.40(m, 1H), 8.17 (s, 1H), 7.90 (ddd, J = 8.0, 3.8, 1.7 Hz, 1H), 7.68 – 7.54 (m,2H), 7.23 (d, J = 8.2 Hz, 1H), 6.72 (d, J = 4.4 Hz, 1H), 6.00 – 5.73 (m, 1H), 4.78 – 4.63 (m, 1H), 3.61 – 3.47 (m, 2H), 3.26 (h, J = 6.9 Hz, 1H), 3.14 –3.06 (m, 2H), 2.81 (q, J = 3.8 Hz, 2H), 2.78 – 2.71 (m, 4H), 2.67 (s, 1H),2.43 (q, J = 12.7 Hz, 1H), 2.02 (d, J = 8.9 Hz, 1H), 1.39 (dd, J = 6.2, 1.5Hz, 6H), 1.32 (dd, J = 6.9, 2.5 Hz, 6H). HRMS (ESI) calcd.for C 30 H 37 ClN6O4S613.2358 [M+H] + Found 613.2350.

[0177] Example 6: Synthesis of compound W-3083

[0178]

[0179] In the first step, following a similar synthetic method to step 2 of W-3071, 1-bromo-5-isopropoxy-2-methyl-4-nitrobenzene (400 mg, 1.46 mmol) was reacted with N-methylpyrrole-3-boronic acid pinacol ester (332 mg, 1.61 mmol) to obtain a yellow oily liquid (378 mg, 94.5%). 1H NMR (400 MHz, DMSO) δ 7.72 (s, 1H), 7.20 – 7.15 (m, 2H), 6.82 (t, J = 2.5 Hz, 1H), 6.43 – 6.38 (m, 1H), 4.84 (dq, J = 12.0, 6.0 Hz, 1H), 3.68 (s, 3H), 2.37 (s, 3H), 1.29 (d, J = 6.0 Hz, 6H).

[0180] The second step involved a synthesis method similar to step 3 of the W-3071 synthesis, yielding a yellow oily liquid (299 mg, 88.7%). 1 H NMR (400 MHz, DMSO) δ 6.73 – 6.69 (m, 2H), 6.66 (t, J = 2.4 Hz,1H), 6.48 (d, J = 0.7 Hz, 1H), 6.09 (dd, J = 2.7, 1.8 Hz, 1H), 4.46 – 4.37(m, 3H), 3.61 (s, 3H), 2.18 (s, 3H), 1.25 (d, J = 6.1 Hz, 6H).

[0181] The third step involved a synthesis method similar to step 4 of the W-3071 synthesis, yielding a yellow solid (151 mg, 33.3%). 1 HNMR (400 MHz, DMSO) δ 9.47 (s, 1H), 8.49 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 8.06 (s, 1H), 7.83 (dd, J = 8.0, 1.6 Hz, 1H), 7.62 (d, J = 9.9 Hz, 2H), 7.34(t, J = 7.6 Hz, 1H), 6.99 – 6.86 (m, 2H), 6.74 (t, J = 2.4 Hz, 1H), 6.23 (t,J = 2.3 Hz, 1H), 4.57 (p, J = 6.1 Hz, 1H), 3.65 (s, 3H), 3.43 (p, J = 6.7 Hz,1H), 2.21 (s, 3H), 1.24 (d, J = 6.1 Hz, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS(ESI) calcd.for C 28 H 32ClN5O3S 554.1987 [M+H] + Found 554.1978.

[0182] Example 7: Synthesis of compound W-3084

[0183]

[0184] The synthesis method of W-3084 is similar to that of W-3071, yielding a yellow solid (33 mg, total yield 3.0%). 1 H NMR (400MHz, DMSO) δ 9.48 (s, 1H), 8.38 (d, J = 8.3 Hz, 1H), 8.31 (d, J = 5.8 Hz, 2H), 8.10 (s, 1H), 7.83 (dd, J = 8.0, 1.6 Hz, 1H), 7.61 – 7.54 (m, 1H), 7.35(td, J = 7.6, 1.2 Hz, 1H), 6.94 (s, 1H), 5.57 (s, 1H), 4.78 (p, J = 6.0 Hz,1H), 3.46 (d, J = 6.7 Hz, 1H), 3.16 (s, 2H), 2.76 (s, 2H), 2.41 (d, J = 10.3Hz, 5H), 1.27 (d, J = 6.0 Hz, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS (ESI)calcd.for C 29 H 33 ClF3N5O3S 624.2017 [M+H] + , found 624.1996.

[0185] Example 8: Synthesis of compounds W-3086 to W-3128

[0186]

[0187] The synthesis method of W-3086 is similar to that of W-2082, yielding a yellow solid (100 mg, 95.4%). 1H NMR (400 MHz, DMSO) δ 9.46 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.24 (d, J = 2.3 Hz, 1H), 8.05 (s, 1H), 7.83 (dd, J = 8.0, 1.7 Hz, 1H), 7.62 (t, J = 7.9 Hz, 1H), 7.54(s, 1H), 7.35 (t, J = 7.6 Hz, 1H), 6.87 (d, J = 2.4 Hz, 1H), 4.61 (p, J = 5.9Hz, 1H), 3.49 – 3.41 (m, 3H), 3.20 (td, J = 12.3, 3.0 Hz, HRMS (ESI) calcd.for C 29 H 35 ClN6O3S 583.2253 [M+H] + , found 583.2233.

[0188]

[0189] The synthesis method of W-3116 is similar to that of W-2082, yielding a white solid (51 mg, 59.5%). 1H NMR (400 MHz, DMSO) δ 9.46 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.24 (s, 1H), 8.07 (s, 1H), 7.83 (dd, J = 8.0, 1.6 Hz, 1H), 7.66 – 7.59 (m, 1H), 7.52 (s, 1H), 7.39 –7.31 (m, 1H), 6.82 (s, 1H), 4.53 (hept, J = 6.1 Hz, 1H), 3.42 (d, J = 6.8 Hz,2H), 3.10 (s, 2H), 2.78 – 2.68 (m, 1H), 2.45 (s, 1H), 2.12 (s, 3H), 1.86 –1.69 (m, 4H), 1.23 (s, 3H), 1.21 (s, 3H), 1.15 (d, J = 6.8 Hz, 6H). HRMS(ESI) calcd.for C 29 H 35 D3ClN5O3S 575.2645 [M+H] + Found 575.2630.

[0190]

[0191] The synthesis method of W-3128 is similar to that of W-2082, yielding a white solid (78 mg, 88.1%). 1 H NMR (400 MHz, DMSO) δ 9.47 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 8.10 (s, 1H), 7.84 (d, J = 7.9 Hz, 1H), 7.62 (t, J = 7.9 Hz, 1H), 7.52 (s, 1H), 7.35 (t, J= 7.8 Hz, 1H), 6.83 (s, 1H), 4.61 – 4.38 (m, 1H), 3.79 (s, 2H), 2.89 (s, 3H), 2.14 (s, 3H), 2.06 – 1.74 (m, 5H), 1.23 (d, J = 6.1 Hz, 6H), 1.16 (d, J = 6.7Hz, 6H). HRMS (ESI) calcd.for C 30 H 35D5ClN5O3S 591.2927 [M+H] + Found 591.2925.

[0192] Example 9: Synthesis of compound W-3129

[0193]

[0194] The synthesis method of W-3129 is similar to that of W-3040, yielding a white solid (47 mg, 53.5%). 1 H NMR (400 MHz, DMSO) δ 9.47 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 8.05 (d, J =9.7 Hz, 2H), 7.87 – 7.81 (m, 1H), 7.63 (t, J = 7.9 Hz, 1H), 7.52 (s, 1H), 7.36 (t, J = 7.6 Hz, 1H), 6.84 (s, 1H), 4.58 (p, J = 6.0 Hz, 1H), 4.37 – 4.30(m, 1H), 3.79 (d, J = 12.9 Hz, 1H), 3.44 (p, J = 6.7 Hz, 1H), 3.23 – 3.12 (m,1H), 2.99 – 2.89 (m, 1H), 2.70 (m, 1H), 2.15 (s, 3H), 1.71 (t, J = 13.7 Hz,2H), 1.52 (m, 2H), 1.20 (d, J = 6.0 Hz, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS(ESI) calcd.for C 29 H 36 ClN5O4S 586.2249 [M+H] + , found 586.2246.

[0195] Example 10: Synthesis of compound W-3132

[0196]

[0197] Step 1: Following the synthesis method of ZJL-4132, a yellow solid (153 mg, 64.3%) was obtained.

[0198] Step 2: Dissolve the yellow solid obtained in Step 1 in DCM (3 ml), add 1,4-dioxane hydrochloric acid solution (3 ml), react for 2 h, evaporate the solvent under reduced pressure and proceed directly to the next step.

[0199] Step 3: Following the synthesis method of ZJL-4132, a yellow solid (66 mg, 24.4%) was obtained. 1 H NMR (400MHz, DMSO) δ 9.46 (s, 1H), 9.30 (m, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.24 (s,1H), 8.06 (d, J = 4.3 Hz, 1H), 7.83 (dd, J = 7.9, 1.6 Hz, 1H), 7.62 (q, J =7.7 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.40 – 7.30 (m, 1H), 6.78 (d, J = 15.6Hz, 1H), 6.46 – 6.12 (m, 1H), 4.98 (q, J = 7.3 Hz, 1H), 4.53 (m,2H), 4.08 –3.94 (m, 1H), 3.44 (q, J = 6.7 Hz, 1H), 3.18 (m, 1H), 3.03 – 2.92 (m, 1H),2.90 – 2.84 (m, 1H), 2.78 – 2.57 (m, 3H), 2.15 (s, 3H), 1.74 – 1.64 (m, 2H),1.45 (m, 1H), 1.37 (m, 1H), 1.22 – 1.19 (m, 6H), 1.16 (d, J = 6.8 Hz, 6H).HRMS (ESI) calcd.for C 35 H 41 ClF2N6O5S 731.2588 [M+H] + , found 731.2573.

[0200] Example 11: Synthesis of compound W-3137

[0201]

[0202] In the first step, 2-bromo-4-fluoro-5-nitrotoluene (400 mg, 1.71 mmol), cyclopropanol (298 mg, 5.13 mmol), cesium carbonate (1114 mg, 3.42 mmol), and THF (6 ml) were added to a 25 ml round-bottom flask and reacted overnight at 60 °C. The reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated, and the product was extracted with 20 ml of ethyl acetate. The organic phase was washed three times with 30 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 100:1) to give a yellow solid (428 mg, 92.0%). 1 H NMR (400 MHz, DMSO) δ 7.93 (d, J = 0.8 Hz, 1H), 7.81(s, 1H), 4.14 (tt, J = 6.0, 2.9 Hz, 1H), 2.34 (d, J = 0.6 Hz, 3H), 0.87 –0.83 (m, 2H), 0.75 – 0.70 (m, 2H).

[0203] The remaining steps were similar to those for the synthesis of W-3071, yielding a white solid (51 mg, total yield 5.2%). 1 H NMR(400 MHz, DMSO) δ 9.49 (s, 1H), 8.49 (d, J = 8.4 Hz, 1H), 8.25 – 8.14 (m,2H), 7.82 (dd, J = 7.9, 1.5 Hz, 1H), 7.58 (t, J = 8.0 Hz, 1H), 7.42 (s, 1H),7.33 (t, J = 7.6 Hz, 1H), 7.13 (s, 1H), 3.82 (dq, J = 6.1, 3.0 Hz, 1H), 3.43(s, 1H), 3.08 (d, J = 11.3 Hz, 2H), 2.77 – 2.68 (m, 1H), 2.38 (d, J = HRMS (ESI) calcd.forC 29 H 36 ClN5O3S 570.2300 [M+H] +, found 570.2291.

[0204] Example 12: Synthesis of compound W-3141

[0205]

[0206] The synthesis method of W-3141 is similar to that of W-2082, yielding a white solid (30 mg, 33.4%). 1 H NMR (400 MHz, DMSO) δ 9.47 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 8.12 (s, 1H), 7.84 (dd, J = 8.1, 1.7 Hz, 1H), 7.62 (t, J = 7.6 Hz, 1H), 7.52 (s, 1H), 7.35(t, J = 7.6 Hz, 1H), 6.85 (s, 1H), 4.50 (p, J = 6.1 Hz, 1H), 3.44 (p, J = 6.8Hz, 4H), 3.00 (m, 3H), 2.15 (s, 3H), 2.04 (m, 2H), 1.86 (d, J = 13.5 Hz, 2H), 1.27 (d, J = 6.6 Hz, 6H), 1.22 (s, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS (ESI)calcd.for C 31 H 42 ClN5O3S 600.2770 [M+H] + Found 600.2767.

[0207] Example 13: Synthesis of compound W-3158

[0208]

[0209] The synthesis method of W-3158 is similar to that of ZJL-4132, yielding a white solid (87 mg, 96.8%). 1H NMR (400MHz, DMSO) δ 9.46 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.24 (s, 1H), 8.05 (s,1H), 7.84 (dd, J = 8.0, 1.6 Hz, 1H), 7.63 (t, J = 8.0 Hz, 1H), 7.52 (s, 1H),7.39 – 7.32 (m, 1H), 6.84 (s, 1H), 4.57 (dt, J = 11.1, 5.4 Hz, 2H), 3.93 (d,J = 13.5 Hz, 1H), 3.14 (t, J = 11.9 Hz, 1H), 2.89 (s, 1H), 2.69 (s, 2H), 2.61 (d, J = 12.4 Hz, 2H), 2.15 (s, 3H), 2.04 (s, 3H), 1.66 (d, J = 12.9 Hz, 3H), 1.20 (d, J = 6.0 Hz, 6H), 1.16 (d, J = 6.8 Hz, 6H). HRMS (ESI) calcd.forC 30 H 38 ClN5O4S 600.24O6 [M+H] + Found 600.2389.

[0210] Example 14: Synthesis of compound W-4006

[0211]

[0212] The synthesis method of W-4006 is similar to that of W-3071, yielding a white solid (133 mg, total yield 22.4%). 1H NMR(400 MHz, DMSO) δ 9.46 (s, 1H), 8.46 (d, J = 8.4 Hz, 1H), 8.25 (s, 1H), 8.04(s, 1H), 7.84 (dd, J = 8.0, 1.6 Hz, 1H), 7.68 – 7.58 (m, 1H), 7.55 (s, 1H), 7.36 (ddd, J = 8.3, 7.4, 1.2 Hz, 1H), 6.82 (s, 1H), 4.58 (hept, J = 6.0 Hz, 1H), 3.44 (p, J = 6.8 Hz, 1H), 2.83 (t, J = 12.1 Hz, 1H), 2.14 (s, HRMS (ESI) calcd.for C 29 H 35 ClF₂N₄O₃S 593.2159 [M+H] + , found593.2164.

[0213] Example 15: Synthesis of compound W-4007

[0214]

[0215] In the first step, 1-bromo-5-isopropoxy-4-nitro-2-(trifluoromethyl)benzene (530 mg, 1.62 mmol), N-Boc-piperazine (362 mg, 1.94 mmol), palladium acetate (18 mg, 0.08 mmol), potassium tert-butoxide (254 mg, 2.27 mmol), BINAP (50 mg, 0.08 mmol), and anhydrous toluene (10 ml) were added to a 50 ml round-bottom flask. The mixture was reacted overnight at 100 °C under an argon atmosphere. The reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated, the mixture was extracted with 30 ml of ethyl acetate, washed three times with 40 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 10:1-20:1) to give a yellow solid (175 mg, 25.0%). 1H NMR (400 MHz, DMSO) δ 8.16 (s,1H), 7.26 (s, 1H), 5.03 (h, J = 6.0 Hz, 1H), 3.46 (t, J = 5.0 Hz, 4H), 2.98(t, J = 4.9 Hz, 4H), 1.42 (s, 9H), 1.31 (d, J = 6.0 Hz, 6H).

[0216] The subsequent steps followed the synthesis method of W-3071, yielding a yellow solid (12 mg, total yield 0.8%). 1 H NMR(400 MHz, CDCl3) δ 9.53 (s, 1H), 8.50 – 8.36 (m, 2H), 8.18 (d, J = 4.3 Hz, 1H), 7.92 (dd, J = 8.0, 1.7 Hz, 1H), 7.61 – 7.50 (m, 2H), 7.37 – 7.28 (m,1H), 6.92 (s, 1H), 4.65 (p, J = 6.1 Hz, 1H), 3.25 (p, J = 6.9 Hz, 1H), 3.02(d, J = 5.8 Hz, 3H), 2.78 (s, 2H), 2.51 (s, 2H), 2.22 (m, 2H), 2.04 (d, J =6.7 Hz, 2H), 1.32 – 1.28 (m, 12H).HRMS (ESI) calcd.for C 28 H 34 ClF3N6O3S 627.2126[M+H] + , found 627.2117.

[0217] Example 16: Synthesis of compound LCX-1

[0218]

[0219] In the first step, 2-bromo-4-fluoro-5-nitrotoluene (1500 mg, 6.41 mmol), 60% sodium hydride (512 mg, 12.82 mmol), and THF (10 ml) were added to a 50 ml double-necked flask at 0 °C and stirred for 30 min. Then, 1,1,1,3,3,3-hexafluoro-2-propanol (1616 mg, 9.62 mmol) was slowly added, and the mixture was allowed to react at room temperature. The reaction was monitored by TLC. After the reaction was completed, the mixture was quenched with ice-cold ammonium chloride solution, extracted with 50 ml of ethyl acetate, washed three times with 70 ml of water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The product was purified by silica gel column chromatography (PE:EA = 60:1-80:1) to obtain a yellow solid (1891 mg, 77.4%). 1 H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 0.9 Hz, 1H), 8.00(s, 1H), 6.84 - 6.78 (m, 1H), 2.38 (s, 3H).

[0220] The remaining steps were similar to those for the synthesis of W-3071, yielding a yellow solid (101 mg, total yield 13.1%). 1 H NMR(400 MHz, Chloroform-d) δ 9.88 (s, 1H), 8.59 (dd, J = 8.4, 1.1 Hz, 1H), 8.22(s, 1H), 7.90 (dd, J = 8.0, 1.6 Hz, 1H), 7.66 – 7.53 (m, 2H), 7.31 – 7.26 (m,1H), 6.99 (s, 1H), 3.26 – 3.17 (m, 3H), 2.76-2.68 (m, 1H), 2.58 (q, J = 7.2Hz, 2H), 2.33 (s, 3H), 2.19 (t, J = 10.8 Hz, 2H), 1.93 – 1.76 (m, 4H), 1.30(d, J = 6.8 Hz, 6H), 1.19 (t, J = 7.2 Hz, 3H). HRMS (ESI) calcd.forC 30 H 34 ClF6N5O3S 694.2048 [M+H] + , found694.2047.

[0221] Example 17: Synthesis of compound LCX-2

[0222]

[0223] The synthesis method of LCX-2 is similar to that of LCX-1, yielding a yellow solid (32 mg, total yield 4.3%). 1 H NMR (400MHz, Chloroform-d) δ 9.90 (s, 1H), 8.60 (d, J = 8.4 Hz, 1H), 8.22 (s, 1H), 7.90 (dd, J = 8.0, 1.6 Hz, 1H), 7.70 – 7.52 (m, 2H), 7.29 (d, J = 7.6 Hz,1H), 6.98 (s, 1H), 3.30 – 3.16 (m, 3H), 2.79 – 2.69 (m, 2H), 2.51 (s, 3H),2.41 – 2.37 (m, 1H), 2.33 (s, 3H), 2.03 – 1.95 (m, 2H), 1.82 (d, J = 13.5 Hz,2H), 1.29 (d, J = 4.4 Hz, 6H). HRMS (ESI) calcd.for C 29 H 32 ClF6N5O3S 680.1891 [M+H] + , found 680.1889.

[0224] Example 18: Synthesis of compound LCX-3

[0225]

[0226] The synthesis method of LCX-3 is similar to that of LCX-1, yielding a yellow solid (84 mg, total yield 17.2%). 1H NMR (600MHz, Chloroform-d) δ 9.54 (s, 1H), 8.62 – 8.52 (m, 1H), 8.16 (s, 1H), 8.05(s, 1H), 7.91 (dd, J = 8.0, 1.6 Hz, 1H), 7.60 (ddd, J = 8.7, 7.3, 1.7 Hz,1H), 7.39 (s, 1H), 7.26 – 7.22 (m, 1H), 6.91 (s, 1H), 4.68-4.62 (m, 1H), 3.35(d, J = 11.5 Hz, 2H), 3.25 (hept, J = 6.8 Hz, 1H), 2.79 (dt, J = 7.5, 3.6 Hz, 1H), 2.75 (q, J = 7.6, 5.9 Hz, 2H), 2.36 (t, J = 11.9 Hz, 2H), 2.20 (s, 3H), 2.10 (t, J = 13.3 Hz, 2H), 1.87 – 1.81 (m, 2H), 1.54 (d, J = 6.5 Hz, 3H), 1.31 (dd, J = 6.9, 5.6 Hz, 6H), 1.30 – 1.27 (m, 3H). HRMS (ESI) calcd.forC 30 H 37 ClF3N5O3S 640.2330 [M+H] + , found 640.2312.

[0227] Example 19: Synthesis of compound LCX-4

[0228]

[0229] The synthesis method of LCX-4 is similar to that of LCX-1, yielding a yellow solid (86 mg, overall yield 9.0%). 1H NMR (600MHz, Chloroform-d) δ 9.55 (s, 1H), 8.58 (d, J = 8.4 Hz, 1H), 8.16 (s, 1H), 8.05 (s, 1H), 7.91 (dd, J = 8.0, 1.6 Hz, 1H), 7.64 – 7.58 (m, 1H), 7.39 (s,1H), 7.24 (d, J = 7.6 Hz, 1H), 6.89 (s, 1H), 4.63 (m, 1H), 3.29 – 3.22 (m,3H), 2.76 (tt, J = 12.2, 3.8 Hz, 1H), 2.52 (s, 3H), 2.45 – 2.35 (m, HRMS (ESI) calcd.for C 29 H 35 ClF3N5O3S626.2174 [M+H] + , found 626.2154.

[0230] Example 20: Synthesis of compound LCX-5

[0231]

[0232] The synthesis method of LCX-5 is similar to that of LCX-1, yielding a yellow solid (48 mg, total yield 7.2%). 1H NMR (400MHz, Chloroform-d) δ 9.53 (s, 1H), 8.56 (d, J = 8.3 Hz, 1H), 8.16 (s, 1H), 8.01 (s, 1H), 7.92 (dd, J = 8.0, 1.6 Hz, 1H), 7.66 – 7.54 (m, 1H), 7.37 (s,1H), 7.24 (d, J = 7.7 Hz, 1H), 6.67 (s, 1H), 4.73-4.66 (m, 1H), 3.37 – 3.28(m, 2H), 3.27 – 3.21 (m, 1H), 3.19 – 3.12 (m, 2H), 2.80 (dd, J = 13.0, 4.5Hz, 2H), 2.74 (d, J = 4.8 Hz, 1H), 2.69 (t, J = 7.2 Hz, 2H), 2.31 (t, J =11.9 Hz, 2H), 2.17 (s, 3H), 2.10 – 2.00 (m, 2H), 1.82 (d, J = 13.3 Hz, 2H), 1.31 (d, J = 6.9 Hz, 6H), 1.29 – 1.27 (m, 3H). HRMS (ESI) calcd.forC 31 H 38 ClF2N5O3S 634.2425 [M+H] + , found 634.2404.

[0233] Example 21: Synthesis of compound LCX-6

[0234]

[0235] The synthesis method of LCX-6 is similar to that of LCX-1, yielding a yellow solid (49 mg, total yield 3.7%). 1H NMR (400MHz, Chloroform-d) δ 9.53 (s, 1H), 8.61 – 8.47 (m, 1H), 8.16 (s, 1H), 8.05(s, 1H), 7.92 (dd, J = 8.0, 1.6 Hz, 1H), 7.61 (ddd, J = 8.7, 7.3, 1.7 Hz,1H), 7.41 (s, 1H), 7.26 – 7.22 (m, 1H), 6.77 (s, 1H), 4.86 – 4.71 (m, 1H),3.65 (d, J = 11.7 Hz, 2H), 3.37 – 3.27 (m,, 2H), 3.24 (q, J = 6.8 Hz, 1H),3.11 – 3.02 (m, 1H), 3.03 – 2.95 (m, 2H), 2.82 (s, 3H), 2.77 – 2.69 (m,, 4H),2.18 (s, 3H), 1.95 (d, J = 14.0 Hz, 2H), 1.31 (d, J = 6.9 Hz, 6H). HRMS (ESI)calcd.for C 30 H 36 ClF2N5O3S 620.2268 [M+H] + , found 620.2249.

[0236] The experimental instruments and materials used in the following embodiments are described below:

[0237] Experimental reagents: CCK8 (K1018, APExBIO, USA), protease inhibitors and phosphatase inhibitors (P1046, Beyotime, CHN), BCA protein assay kit (P0009, Beyotime, CHN), BSA (9048-46-8, MPBiomedicals LLC, USA), StarSignal Plus chemiluminescence assay kit (E170-102, Genstar, CHN), RIPA lysis buffer (89900, Thermo Fisher, USA), light / heavy isotope-labeled azide biotin (EVU102 / EVU151, KeraFast, USA), sodium ascorbate (A7631, Sigma-Aldrich, USA), TBTA (678937, Sigma-Aldrich, USA), copper sulfate (209198, Sigma-Aldrich, USA), streptavidin agarose beads (20359, Thermo Fisher, USA), Protein A+G Affinity beads (88802, Thermo Fisher, USA), Amaxa Lonza Cell Line Nucleofector® Kit V transfection kit (VCA-1003, Cologne, GER), G418 (A1720-1G, Merck, USA); primary antibodies against ALK (3633S), FAK (3285S), P-AKT (4060S), AKT (4691S), P-ERK1 / 2 (9101S), and ERK1 / 2 (4695S) were purchased from Cell Signaling Technology. β-actin (AA128-1) antigen-specific antibody was purchased from Beyotime.

[0238] Experimental instruments: Ultrasonic cell disruptor (SCIENTZ, CHN), Synergy H1 hybrid multi-functional microplate reader (BioTek, USA), dry thermostat (ALLSHENG, CHN), ultra-sensitive multi-functional imaging system (General Electric Company, USA), flow cytometer (Merck Millipore, DE).

[0239] Example 22 Inhibitory activity of compounds against the proliferation of H3122, Ba / F3-EML4-ALK and their mutant cells

[0240] The H3122 (human non-small cell lung cancer cells) and Ba / F3-EML4-ALK (a stable ALK fusion protein model cell) used in this experiment were purchased from the Shanghai Cell Bank. Ba / F3-EML4-ALK cells were constructed using the following standard methods. L1196M and Ba / F3-EML4-ALK G1269A : EML4-ALK L119M Or EML4-ALK G1269A The plasmid pcDNA3.1 was cloned and transfected into Ba / F3 cells via electroporation using the AmaxaLonza Cell Line Nucleofector® Kit V. Subsequently, cells stably expressing the plasmid were selected by adding G418, and interleukin-3 was withdrawn before culturing to obtain Ba / F3-EML4-ALK. L1196M (Model cells stably expressing the drug-resistant mutant of the ALK fusion protein L1196M) and Ba / F3-EML4-ALK G1269A (Model cells stably expressing the drug-resistant mutant of the ALK fusion protein G1269A).

[0241] H3122, Ba / F3-EML4-ALK, and their mutant cells, in logarithmic growth phase, were seeded into 96-well plates and cultured overnight in an incubator. Then, different concentrations of the compound were added to the corresponding wells of the 96-well plates to achieve a final concentration of 0 μM–10 μM, while maintaining the DMSO concentration at 0.1%. After 72 hours of incubation, 10 μL of CCK-8 reagent was added to each well, and incubation continued for another 2 hours. Subsequently, the OD was measured using a microplate reader. 450 and OD 650 Cell viability V% = (As − Ac) / (Ab − Ac) × 100%, where As is the absorbance of the experimental group, Ab is the absorbance of the control group (using DMSO as a control), and Ac is the absorbance of the blank group (cell-free group). 50 The values ​​were calculated using GraphPad Prism 8.0 software.

[0242] The test results are shown in Tables 1 and 2: The compounds of this invention exhibit excellent inhibitory activity against the proliferation of H3122 and Ba / F3-EML4-ALK cells. The molecular gel compounds ZJL-4155, W-2082, W-2083, and ZJL-5055 of this invention all show strong inhibitory activity against the proliferation of H3122, Ba / F3-EML4-ALK, and their mutant cells. The marketed drug Crizotinib has good inhibitory activity against ALK-driven tumor cells such as H3122 and Ba / F3-EML4-ALK, but not against Ba / F3-EML4-ALK cells. L1196M Ba / F3-EML4-ALK G1269A The inhibitory activity against the proliferation of mutant drug-resistant cells was significantly reduced. The compounds of this invention retained their inhibitory activity against mutant drug-resistant cells, demonstrating significant superiority over the marketed drug Crizotinib.

[0243] Table 1. IC50 (µM) of the inhibitory activity of the compounds on the proliferation of H3122 and Ba / F3-EML4-ALK cells.

[0244]

[0245]

[0246] Table 2. IC50 (µM) of the inhibitory activity of the compounds on the proliferation of H3122, Ba / F3-EML4-ALK and their mutant cells.

[0247]

[0248] Example 23: Degradation activity of compounds against ALK and FAK in non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells.

[0249] H3122 and Ba / F3-EML4-ALK cells (2×10⁻⁶) were used. 5Cells were seeded into 6-well plates and incubated overnight. The next day, cells were treated with different concentrations of compounds for 16 hours, washed twice with PBS buffer, and all cells were collected using RIPA lysis buffer containing 1× protease inhibitor and phosphatase inhibitor. Cells were lysed using an ultrasonic cell disruptor (3 seconds on, 3 seconds off, 12% power, repeated 3 times). Protein concentration was determined using a BCA protein assay kit and a multi-mode microplate reader. The protein samples were then heated at 98°C for 10 minutes using a dry thermostat. After separation by SDS-polyacrylamide gel electrophoresis, proteins were transferred to PVDF membranes using wet transfer. The PVDF membranes were then blocked for 1 hour at room temperature with 1×TBST buffer containing 5% BSA. After blocking, corresponding PVDF membrane strips were cut and incubated overnight at 4°C with the corresponding primary antibody. After incubation, the PVDF membranes were washed three times with 1×TBST buffer (10 min - 5 min - 10 min). Then, add the corresponding secondary antibody and incubate for 1 hour. Wash the PVDF membrane three times with 1×TBST buffer (10 min - 5 min - 10 min) to remove unbound secondary antibody and reduce background signal. After washing, gently blot the PVDF membrane dry with absorbent paper, place it on a tray, and evenly add freshly prepared ECL chemiluminescence solution (solution A:solution B = 1:1), ensuring that the chemiluminescence solution completely covers the PVDF membrane. Expose and develop using an ultrasensitive multi-functional imager.

[0250] like Figures 1-3 As shown in Table 3, compounds ZJL-4155, ZJL5055, and W-2082 exhibited potent degradative activity against ALK and FAK proteins at a concentration of 100 nM. Furthermore, ZJL-4149, ZJL-4151, ZJL-4152, ZJL-4153, W-2083, W-3116, W-3128, W-3129, W-3132, and W-3158 also showed strong degradative activity. Ceritinib showed no degradative activity against either ALK or FAK proteins.

[0251] Table 3. Degradation activity of compounds against ALK protein in H3122 cells.

[0252]

[0253] Example 24: Degradation of ALK and FAK proteins by compound ZJL-4155 in non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells.

[0254] Concentration-dependent and time-dependent experiments were performed following the experimental methods described in Example 23. For the concentration-dependent experiments, non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells were seeded in 6-well plates and treated with different concentrations of ZJL-4155 (0, 6.25, 12.5, 25, 50, and 100 nM) for 16 hours, followed by Western blotting (WB) analysis. For the time-dependent experiments, cells were treated with the same concentration of ZJL4155 (50 nM) at different time points (0, 2, 4, 8, 16, and 24 hours) before WB analysis.

[0255] Concentration-dependent experiments showed that compound ZJL-4155 effectively degraded ALK and FAK proteins in H3122 cells, and its DC... 50 The concentrations were 15.96 nM and 14.52 nM, respectively, with Dmax values ​​of 89.75% and 88.11%, respectively. Furthermore, compound ZJ-L4155 can also effectively degrade ALK in Ba / F3-EML4-ALK cells, with a DC value of [missing value]. 50 The concentration was 27.77 nM, and the Dmax was 88.65%. Time-dependent experiments showed that compound ZJL4155 effectively degraded ALK and FAK proteins after 16 hours of incubation in H3122 cells, and effectively degraded ALK protein after 16 hours of incubation in Ba / F3-EML4-ALK cells. Figure 4 ).

[0256] In addition to the time-dependent assay, an elution assay was performed. H3122 cells and Ba / F3-EML4-ALK cells were treated with compound ZJL-4155 (50 nM) for 16 hours, then washed off the drug and re-cultured in fresh culture medium. The results showed that after treatment with compound ZJL-4155 and subsequent elution, the levels of ALK and FAK proteins in H3122 and Ba / F3-EML4-ALK cells did not recover within 24 hours, indicating that compound ZJL-4155 has a sustained degradation capacity. Figure 5 ).

[0257] In addition, the effect of compound ZJL-4155 on the downstream ALK signaling pathway was tested using the experimental method described in Example 23. H3122 cells were seeded in 6-well plates and treated with different concentrations of ZJL-4155 (0, 6.25, 12.5, 25, 50, and 100 nM) for 16 hours, followed by Western blotting analysis. The results showed that compound ZJL-4155 at a concentration of 25 nM significantly inhibited the phosphorylation of AKT and ERK1 / 2, and its inhibitory effect was superior to that of Ceritinib (…). Figure 6 ).

[0258] The above results demonstrate that ZJ-L4155 is a potent ALK and FAK molecular gel degrader that can effectively inhibit the transmission of their cancer signals.

[0259] Example 25 Compound ZJL-4155 can degrade ALK and FAK proteins via the ubiquitin-proteasome pathway.

[0260] (1) Whole-cell proteomics verification of the degradation of ALK and FAK proteins mediated by compound ZJL-4155: H3122 cells were treated with compound ZJL-4155 (100 nM) for 16 hours, and the cells were collected, lysed, and the protein concentration was quantified by BCA. 1 mg of total protein was dissolved in 1X TEAB buffer, reduced with DTT, alkylated with IAA, and digested with trypsin at 37°C overnight. Peptides were collected by centrifugation, desalted on a C18 column, vacuum dried, and then analyzed by LC-MS / MS. Protein identification and label-free quantification were performed using MaxQuant software, and differential protein analysis was performed using Perseus software. The results showed that the abundance of ALK and FAK proteins was significantly reduced, confirming the selective clearance of ALK and FAK proteins. Figure 7 ).

[0261] (2) Investigating the degradation mechanism of compound ZJL-4155: H3122 cells were pretreated with the proteasome inhibitor MG132 (100 nM) and the lysosome inhibitor BafA1 (100 nM) for 2 hours, and then treated with compound ZJL-4155 for 16 hours. Cells were then lysed for Western blot analysis. The results showed that MG132 could block the degradation of ALK and FAK proteins mediated by compound ZJL-4155, while BafA1 had no such effect. Figure 8 ).

[0262] (3) Detection of protein ubiquitination: H3122 cells were pretreated with MG132 (100 nM) for 2 hours, followed by treatment with compound ZJL-4155 (100 nM) for 6 hours. After cell lysis, a protein solution was obtained, and 5 μL of primary antibody ALK was added. After incubation for 4 hours, Protein A / G beads were added for enrichment overnight. The next day, after washing the Protein A / G beads, 50 μL of 2 × SDS loading buffer was added, and the mixture was heated at 98°C for 30 minutes. The protein sample was used for subsequent Western blotting to detect ubiquitination levels. The results showed that after pre-incubation with MG132 to block the ubiquitin-proteasome pathway, compound ZJL-4155 could induce potent ubiquitination of ALK protein in H3122 cells. Figure 8 ).

[0263] All of the above results demonstrate that compound ZJL-4155 induces the degradation of ALK and FAK proteins via the ubiquitin-proteasome pathway.

[0264] Example 26: Compound ZJL-4155 can bind to the Cys232 site of TRIM28.

[0265] H3122 cells were treated with ZJL4155 (100 nM) for 6 hours, then lysed. Iodoacetamide-alkyne (IA-alkyne) probe was added to a final concentration of 50 μM, and the cells were incubated at 37°C in a Ferris wheel for 2 hours. Then, DTT was added for reduction, followed by IAA alkylation, and protein was precipitated using a methanol-chloroform system. The precipitated protein was reconstituted in 50 mM ammonium bicarbonate solution, quantified using BCA, and 1 mg of protein was taken and trypsin was added at an enzyme:protein ratio of 1:50. The cells were then enzymatically digested at 37°C for 16 hours. After desalting the enzymatic hydrolysis product via an HLB extraction column, 30 μL of a solution containing 30% acetonitrile was added, followed by a click reaction: 1 mM light / heavy isotope-labeled azide-UV-biotin (1 μL each in 40 mM stock solution), 10 mM sodium ascorbate (4 μL in 100 mM stock solution), 1 mM TBTA (1 μL in 50 mM stock solution), and 10 mM copper sulfate (4 μL in 100 mM stock solution) were added. After incubation at room temperature in the dark for 2 hours, the light and heavy isotope-labeled samples were immediately mixed. Subsequently, the peptides were diluted with 120 μL of 10×50 mM sodium acetate buffer, streptavidin agarose beads were added, and after incubation at room temperature for 2 hours, the magnetic beads were washed. The magnetic beads were resuspended in 25 mM ammonium bicarbonate solution and photolyzed by irradiation with a 365 nm UV lamp at room temperature for 2 hours. The supernatant was collected, concentrated under vacuum, and then desalted via an HLB extraction column. After vacuum drying again, the sample was sent for LC-MS / MS analysis.

[0266] The results showed that the cysteine ​​residue (C232, R=8.09) at TRIM28 was significantly competitive, and LC-MS / MS spectra confirmed covalent modification at this site. Figure 9 These results demonstrate that compound ZJL-4155 induces ALK and FAK protein degradation via the ubiquitin-proteasome pathway through covalent modification of the Cys232 site of TRIM28.

[0267] Next, we further investigated the effects of TRIM28 on ALK and FAK proteins in cells. shRNA was transfected into H3122 cells using RNAiMAX transfection reagent. After incubation for 48 hours in a constant temperature incubator, cells were collected and lysed. The resulting cell lysates were used for subsequent Western blotting analysis to observe whether shRNA could effectively knock out TRIM28 expression in H3122 cells and to observe changes in the expression levels of ALK and FAK proteins.

[0268] The results showed that knocking out TRIM28 significantly upregulated the expression of ALK and FAK proteins. Figure 10 This indicates that the E3 ligase can indeed affect the expression of ALK and FAK proteins, and may be involved in the degradation process of ALK and FAK proteins induced by compound ZJL-4155.

[0269] Example 27: Degradation of ALK and FAK proteins by non-covalent molecular gel W-2082 in non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells

[0270] Concentration-dependent and time-dependent experiments were performed following the experimental methods described in Example 23. For the concentration-dependent experiments, non-small cell lung cancer H3122 and Ba / F3-EML4-ALK cells were seeded in 6-well plates and treated with different concentrations of W-2082 (0, 6.25, 12.5, 25, 50, and 100 nM) for 16 hours, followed by Western blotting (WB) analysis. For the time-dependent experiments, cells were treated with the same concentration of W-2082 (50 nM) at different time points (0, 2, 4, 8, 16, and 24 h) before WB analysis.

[0271] Concentration-dependent experiments showed that compound W-2082 effectively degraded ALK and FAK proteins in non-small cell lung cancer H3122 cells, and its DC... 50 The concentrations were 18.74 nM and 19.32 nM, respectively, with Dmax values ​​of 82.83% and 95.37%, respectively. Furthermore, W-2082 can also effectively degrade ALK in Ba / F3-EML4-ALK cells, with its DC... 50 The concentration was 10.84 nM, and the Dmax was 91.46%. Time-dependent experiments showed that compound W-2082 effectively degraded ALK and FAK proteins after 16 hours of incubation in H3122 cells and Ba / F3-EML4-ALK cells. Figure 11 ).

[0272] In addition to the time-dependent experiments, an elution experiment was also conducted. H3122 cells and Ba / F3-EML4-ALK cells were treated with compound W-2082 (50 nM) for 16 hours, then the drug was washed off and the cells were re-cultured in fresh culture medium. The elution experiment results showed that compound W-2082 at a concentration of 50 nM could persistently degrade ALK and FAK proteins. Figure 12 ).

[0273] The effect of compound W-2082 on the downstream ALK signaling pathway was tested using the Western blot method described in Example 23. H3122 cells were seeded in 6-well plates and treated with different concentrations of ZJL-4155 (0, 6.25, 12.5, 25, 50, and 100 nM) for 16 hours before Western blot analysis. The results showed that compound W-2082 at a concentration of 12.5 nM effectively inhibited the phosphorylation of ALK downstream proteins AKT and ERK1 / 2, and its inhibitory effect was superior to that of Ceritinib. Figure 13 ).

[0274] The above results demonstrate that the non-covalent molecular adhesive W-2082 is also a potent ALK and FAK molecular adhesive degrader, effectively inhibiting the transmission of their cancer signals.

[0275] Example 28: Non-covalent molecular gel W-2082 can degrade ALK and FAK proteins via the ubiquitin-proteasome pathway.

[0276] H3122 cells were pretreated for 2 hours with the proteasome inhibitor MG132 (100 nM), the lysosome inhibitor BafA1 (100 nM), and the autophagy and lysosome inhibitor Chloroquine (100 nM), and then treated with compound W-2082 for 16 hours. After that, the cells were lysed and analyzed by Western blot (WB) according to the method in Example 25.

[0277] The results showed that MG132 could block the degradation of ALK and FAK proteins mediated by compound W-2082, while BafA1 and Chloroquine had no such effect, indicating that the degradation of ALK and FAK proteins by compound W-2082 depends on the ubiquitin-proteasome pathway. Figure 14 ).

[0278] These results indicate that compounds W-2082 and ZJL-4155 have the same degradation mechanism for ALK and FAK proteins.

[0279] Example 29: Compound W-2082 on BaF3-EML4-ALK G1269A Degradation activity and induction of cell cycle arrest and apoptosis

[0280] (1) The effect of compound W-2082 on Ba / F3-EML4-ALK was tested according to the method of Example 27. G1269A ALK in cells G1269A Degradation activity of the mutant. Concentration-dependent experiments showed that compound W-2082 exhibited degradation activity in Ba / F3-EML4-ALK. G1269A Effective degradation of ALK in cells G1269A mutants, their DC 50The concentration was 21.10 nM, and the Dmax was 83.47%. Time-dependent experiments showed that compound W-2082 was effective in Ba / F3-EML4-ALK... G1269A ALK can be effectively degraded by incubating in cells for 16 hours. G1269A mutant ( Figure 15 ).

[0281] (2) Compound W-2082 on Ba / F3-EML4-ALK G1269A Effects on cell cycle: This affects BA / F3-EML4-ALK cells in the logarithmic growth phase. G1269A With 2×10 5 Cells were seeded at a density of 1 / 2 well in 6-well plates and incubated at 37°C in a 5% CO2 incubator. The next day, different concentrations of compound W-2082 were added to the culture medium, and the negative control group was added with an equal volume of DMSO. The plates were then incubated for another 24 hours. Subsequently, the cells were digested and centrifuged, and the resulting cell pellet was fixed overnight with pre-chilled 70% ethanol. The next day, the cells were washed with PBS, centrifuged to obtain the cell pellet, and freshly prepared propidium iodide (PI) staining working solution was added. The cells were gently resuspended by pipetting and incubated in a 37°C water bath in the dark for 30 minutes. The propidium iodide staining working solution consisted of 0.5 mL staining buffer, 25 μL propidium iodide staining solution (20×), and 10 μL RNase A (50×), and was prepared fresh for each use. Afterward, flow cytometry was used to detect the stained cells and analyze the proportions of cells in G0 / G1, S, and G2 / M phases. The results showed that compound W-2082 could induce Ba / F3-EML4-ALK. G1269A Cell cycle arrest occurs in the G0 / G1 phase ( Figure 16 ).

[0282] (3) Compound W-2082 against Ba / F3-EML4-ALK G1269A Effects on apoptosis: This study compared BA / F3-EML4-ALK cells in the logarithmic growth phase. G1269A With 3×10 5Cells were seeded at a density of 1 / 2 well in 6-well plates and incubated at 37°C in a 5% CO2 incubator. The next day, cells were treated with different concentrations of compound W-2082 for 48 hours. After incubation, suspended and adherent cells were collected from the 6-well plates, centrifuged at 1000 rpm for 5 minutes, the supernatant was discarded, and the cells were resuspended in PBS. 200 μL of cell suspension from each group was transferred to 7-AAD and AV single-label tubes. 100 μL of 1X Binging Buffer was added to each group to resuspend the cells. 2.5 μL of Annexin-V and 2.5 μL of 7-AAD were added to each group, while the pre-reserved single-label tubes contained only 2.5 μL of Annexin-V or 2.5 μL of 7-AAD. The cells were incubated at room temperature in the dark for 15 minutes. After 15 minutes, 400 μL of 1X Binging Buffer was added, the cells were transferred to ice, protected from light, and analyzed. The results showed that compound W-2082 could significantly induce BA / F3-EML4-ALK G1269A Apoptosis occurs in cells ( Figure 17 ).

[0283] The above results all demonstrate that compound W-2082 is effective against ALK. G1269A The mutant protein also exhibits potent degradation activity and can effectively induce Ba / F3-EML4-ALK degradation. G1269A Cells undergo G0 / G1 phase arrest and apoptosis.

[0284] Example 30: Pharmacokinetics of compound W-2082

[0285] Six male SPF-grade SD rats were randomly divided into a single intravenous injection group (2 mg / kg) and a single gavage group (10 mg / kg). Blood samples were collected from the orbital venous plexus 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 2500 rpm for 10 minutes at 4°C. The drug concentration in plasma was determined by LC-MS. The plasma drug concentration data were processed using Phoenix WinNonlin 8.1, and the main pharmacokinetic parameters were calculated using a non-compartmental model.

[0286] The results show that ( Figure 18 (See Tables 4 and 5). Following a single intravenous administration, the clearance rate of compound W-2082 was CLz = 10.78 ± 1.35 mL / min / kg, the apparent volume of distribution was Vz = 1784.79 ± 522.30 mL / kg, and the elimination half-life was t. 1 / 2z= 1.91 ± 0.52 h. Following a single oral gavage administration, the compound exhibited excellent pharmacokinetics, with an oral bioavailability of F = 32.74 ± 6.20% and an elimination half-life of t = 1.91 ± 0.52 h. 1 / 2z = 4.03 ± 0.29 h and peak plasma concentration of C max = 428.62 ± 65.90 ng / mL, which meets the requirements for oral administration.

[0287] Table 4. Pharmacokinetic results of a single intravenous administration of compound W-2082

[0288]

[0289] Table 5. Pharmacokinetic results of a single oral administration of compound W-2082

[0290]

[0291] Example 31: In vivo reaction of compound W-2082 against Ba / F3-EML4-ALK and Ba / F3-EML4-ALK L1196M orBa / F3-EML4-ALK G1269A Inhibitory effect of transplanted tumors

[0292] Animal experiments were approved by the institution's animal ethics committee. Male BALB / c-Nude (6 weeks old) were purchased from Guangdong Yaokang Laboratory Animal Technology Co., Ltd. After the quarantine period, Ba / F3-EML4-ALK and Ba / F3-EML4-ALK were administered. L1196M orBa / F3-EML4-ALK G1269A (7×10) 6 (200 μL / tumor) was subcutaneously injected into the right anterior axillary subcutaneous tissue of mice until the tumor volume reached 100 mm². 3 Mice were randomly divided into four groups (n = 5) after being left-right rear-end collisions. Each group received either W-2082 (20 mg / kg, qd; 40 mg / kg, qd), a positive control drug (Ceritinib, 20 mg / kg, qd), or a solvent (0.5% CMC-Na, 0.1 ml / 10g) via gavage. Animal weight and tumor size were measured daily. Tumor volume (V) was calculated as follows: V = L × W × W / 2 (L, tumor length; W, tumor width). Animals were sacrificed on day 12, and tumors were collected via dissection. The tumor growth inhibition value (TGI) was calculated using the following formula: TGI = (V × W × W / 2) / 2. 溶媒 - V 给药 / V 溶媒 ) × 100%, (V 给药 V represents the average tumor volume in the treated group of animals; 溶媒(mean tumor volume of animals in the solvent group)

[0293] The results showed that in the Ba / F3-EML4-ALK xenograft model, compared with the solvent control group, compound W-2082 achieved TGI values ​​of 95.6% and 96.0% at doses of 20 mg / kg and 40 mg / kg, respectively, demonstrating a highly significant tumor-suppressive effect. In contrast, Ceritinib only achieved a TGI value of 40.8% at a dose of 20 mg / kg. Figure 19 ). in Ba / F3-EML4-ALK L1196M In xenograft tumor models, compound W-2082 showed TGI values ​​of 95.2% and 95.9% at doses of 20 mg / kg and 40 mg / kg, respectively, while Ceritinib showed a TGI value of 52.6% at a dose of 20 mg / kg. Figure 20 ). in Ba / F3-EML4-ALK G1269A In xenograft tumor models, compound W-2082 showed TGI values ​​of 98.2% and 98.6% at doses of 20 mg / kg and 40 mg / kg, respectively, while Ceritinib showed a TGI value of 39.5% at a dose of 20 mg / kg. Figure 21 It can be seen that compound W-2082 has in vivo effects on Ba / F3-EML4-ALK and Ba / F3-EML4-ALK. L1196M orBa / F3-EML4-ALK G1269A The tumor-suppressing effect was significantly better than that of Ceritinib. During the treatment period, no significant weight loss, death, or other obvious toxic side effects were observed in any of the treatment groups, indicating that the drug was well tolerated.

[0294] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. 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 modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A pyrimidine diamine compound having the structure shown in Formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. in, R is selected from: R1 substituted or unsubstituted 5-7 membered heterocyclic group, R2 substituted or unsubstituted C5-C7 cycloalkyl group, 5-6 membered heteroaryl group, and C1-C3 alkyl substituted 5-6 membered heteroaryl group; R1 is selected from: , C1-C6 alkyl, deuterated C1-C6 alkyl, cyano; R2 is selected from: C1-C6 alkyl groups, halogens; R3 is selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, 3-4 membered heterocyclic group, 5-10 membered heteroaryl group. , , , , , R6 and R7 are independently selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, cyano, C3-C6 cycloalkyl, R9-substituted benzoyl, and R9-substituted naphthoyl; R9 is selected from: hydrogen, C1-C6 alkoxy, C1-C6 alkyl, and halogen; R8 is selected from: hydrogen and C1-C6 alkyl. R4 is selected from: C1-C6 alkoxy, C3-C6 cycloalkyloxy, halogen-substituted C1-C6 alkoxy, and halogen-substituted C3-C6 cycloalkyloxy. R5 is selected from: hydrogen, C1-C6 alkyl, halogen-substituted C1-C6 alkyl. .

2. The pyrimidine diamine compound or its pharmaceutically acceptable salt or stereoisomer according to claim 1, characterized in that, R4 is selected from: C1-C3 alkoxy, C3-C4 cycloalkyloxy, , Halogen-substituted C3-C4 cycloalkyloxy groups; Preferably, R4 is selected from: methoxy, ethoxy, , , , ; Preferably, R4 is .

3. The pyrimidine diamine compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R5 is selected from: methyl, trifluoromethyl, ; Preferably, R5 is methyl.

4. The pyrimidine diamine compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, The heteroatom on the ring in the R1-substituted or unsubstituted 5-7 membered heterocyclic group is one or two nitrogen atoms; And / or, the heteroatom on the ring in the 5-6-membered heteroaryl or C1-C3 alkyl-substituted 5-6-membered heteroaryl is one or two nitrogen atoms.

5. The pyrimidine diamine compound according to claim 4, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, R is selected from: , ; Preferably, R is selected from: ; More preferably, R is .

6. The pyrimidine diamine compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, R1 is selected from: C1-C6 linear alkyl groups, deuterated C1-C6 linear alkyl groups; R2 is selected from: methyl, ethyl, fluorine, chlorine; Preferably, R1 is selected from: C1-C4 straight-chain alkyl groups, deuterated C1-C3 straight-chain alkyl groups; More preferably, R1 is selected from: , methyl, ethyl, n-butyl, deuterated methyl, deuterated ethyl; Preferably, R2 is fluorine.

7. The pyrimidine diamine compound or its pharmaceutically acceptable salt or stereoisomer according to any one of claims 1-5, characterized in that, R3 is selected from: hydrogen, C1-C3 alkyl, fluorine-substituted C1-C3 alkyl, bromine-substituted C1-C3 alkyl, 3-membered heterocyclic group, 9-membered nitrogen-containing aryl group. , , , , , ; R6 and R7 are independently selected from: hydrogen, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, cyano, C3-C4 cycloalkyl, R9-substituted benzoyl, and R9-substituted naphthoyl. R9 is selected from: hydrogen, C1-C3 alkoxy, C1-C3 alkyl; R8 is selected from: hydrogen, C1-C3 alkyl; Preferably, R9 is selected from: hydrogen, methoxy, ethoxy; Preferably, R8 is selected from: hydrogen, methyl, ethyl.

8. The pyrimidine diamine compound according to claim 7, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, R3 is selected from: hydrogen, methyl, ethyl, difluoromethyl, difluoroethyl. , , , , , , , , , , , ; Preferably, R3 is selected from: hydrogen, methyl, ethyl, difluoromethyl, difluoroethyl, ... , , , , .

9. The pyrimidine diamine compound according to claim 8, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, R1 is selected from: difluoroacetyl, methyl, ethyl, n-butyl.

10. The pyrimidine diamine compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The pyrimidine diamine compounds are selected from the following compounds: 。 11. The use of the pyrimidine diamine compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of ALK inhibitors and / or FAK inhibitors.

12. Use of the pyrimidine diamine compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of ALK protein and / or FAK protein degrading agents.

13. The application according to claim 11 or 12, characterized in that, The ALK is one or more of wild-type ALK, fusion mutant ALK, and point mutant ALK; Preferably, the fusion mutant ALK is selected from one or more of EML4-ALK fusion protein, KIF5B-ALK fusion protein, TFG-ALK fusion protein, and CD74-ALK fusion protein; Preferably, the point mutation type ALK is selected from one or more of L1196M mutation ALK, G1269A mutation ALK, G1202R mutation ALK, F1174C / L mutation ALK and I1171N mutation ALK.

14. The use of the pyrimidine diamine compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of an ALK inhibitor, wherein the ALK inhibitor is capable of inhibiting the protein activity of EML4-ALK fusion protein, the EML4-ALK fusion protein L1196M resistance mutant, and / or the EML4-ALK fusion protein G1269A resistance mutant.

15. The use of the pyrimidine diamine compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of an ALK degrading agent, said ALK degrading agent being capable of degrading EML4-ALK fusion protein, EML4-ALK fusion protein L1196M drug-resistant mutant, and / or EML4-ALK fusion protein G1269A drug-resistant mutant.

16. The use of the pyrimidine diamine compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in a medicament for the treatment and / or prevention of tumors.

17. The application according to claim 16, characterized in that, The tumor is a tumor caused by high ALK expression or abnormal ALK activation; Preferably, the form of ALK aberrant activation is selected from one or more of fusion, point mutation, and amplification; Preferably, the tumor is caused by an ALK fusion mutation and / or an ALK point mutation; Preferably, the ALK fusion is selected from one or more of EML4-ALK fusion, KIF5B-ALK fusion, TFG-ALK fusion, and CD74-ALK fusion; Preferably, the ALK point mutation is selected from one or more of the following: L1196M mutation, G1269A mutation, G1202R mutation, F1174C / L mutation, and I1171N mutation.

18. The application according to claim 17, characterized in that, The tumor is a tumor expressing the EML4-ALK fusion protein; And / or, the tumor is a tumor expressing the drug-resistant mutant of the EML4-ALK fusion protein L1196M; And / or, the tumor is a tumor expressing a drug-resistant mutant of the EML4-ALK fusion protein G1269A.

19. The application according to any one of claims 16-18, characterized in that, The tumor is lung cancer, preferably non-small cell lung cancer.

20. A pharmaceutical composition, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises a pyrimidine diamine compound as described in any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

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