Heterocyclic compounds as bcl-2 inhibitors

Abstract

This invention provides heterocyclic compounds as BCL-2 inhibitors as shown in formula (I), pharmaceutical compositions containing such heterocyclic compounds, and uses of said heterocyclic compounds for treating or preventing BCL-2-mediated diseases and functional disorders, such as tumors. The invention also relates to methods for preparing said heterocyclic compounds.

Description

Technical Field

[0001] This invention relates to heterocyclic compounds, pharmaceutical compositions containing the same, and their use as inhibitors of B-cell lymphoma-2 (BCL-2). More specifically, the invention provides novel heterocyclic compounds as BCL-2 inhibitors, pharmaceutical compositions containing such heterocyclic compounds, and the use of said heterocyclic compounds for the treatment or prevention of BCL-2-mediated diseases and functional disorders, such as tumors. The invention also relates to methods for preparing said heterocyclic compounds. Background Technology

[0002] The BCL-2 protein family is one of the core regulatory mechanisms of apoptosis (also known as programmed cell death). It receives and transmits intrinsic intracellular signals or external environmental stress signals, such as nutrient or hypoxic stress, DNA damage, overactivation of oncogenes, and endoplasmic reticulum stress. It plays a dominant role in the intrinsic apoptotic pathway. The BCL-2 (B-cell lymphoma-2) protein was first discovered in 1986 and is expressed by the BCL-2 gene. The BCL-2 gene is a proto-oncogene, and the proteins it expresses are called BCL-2 family proteins. There are 27 BCL-2 family proteins in the human body, which can be divided into three subclasses based on function and sequence analysis. The first subclass antagonizes apoptosis and includes BCL-XL, BCL-2, BCL-W, MCL-1, and BFL-1. These are mainly located on mitochondria, protecting them from environmental damage. The other two subclasses promote apoptosis; one subclass is the final executor of mitochondrial damage, including BAX and BAK. The rest belong to the BH3 subclass and can directly sense various cellular stress signals. The dynamic balance of interactions between antagonistic and apoptosis-promoting proteins determines the life and death of cells. The apoptosis-antagonistic BCL-2 protein is closely related to tumors; approximately 50% of tumors (such as leukemia, rectal cancer, and prostate cancer) show abnormal overexpression of BCL-2 family proteins, with abnormal BCL-2 activity being widespread in hematological malignancies. Multiple signaling pathways, including JAK-STAT, NFkB, and UPS (ubiquitin-proteasome system), can induce overexpression of apoptosis-antagonistic BCL-2 proteins.

[0003] High expression of BCL-2 family antagonistic apoptosis proteins is associated with drug resistance in various tumors. For example, overexpression of BCL-2 antagonistic apoptosis proteins can enable tumor cells to evade apoptosis induced by antitumor drugs, thereby leading to drug resistance. Studies have shown that inhibiting BCL-2 family proteins can inhibit tumor angiogenesis, thereby inhibiting tumor metastasis (Benjamin, D.; Isaac, J.). et al.J. Clin. Oncol. 2008, 26(25), 4180). Therefore, targeting and inhibiting BCL-2 family anti-apoptotic proteins can suppress tumor development, progression, and drug resistance.

[0004] Although more than 20 small molecule inhibitors targeting the BCL-2 family have been reported, very few have entered clinical trials. Teva's Obatoclax only achieved a partial response in 1 of 26 patients with chronic lymphocytic leukemia (CLL) and exhibited strong neurotoxicity, leading to the termination of development in 2013. AbbVie's Navitoclax (ABT-263), while showing a good response rate of 50% in a phase I dose-escalation trial in patients with relapsed or refractory lymphomas, also exhibited very strong BCL-XL targeting toxicity, such as thrombocytopenia and severe anemia. Venetoclax (ABT-199), jointly developed by AbbVie and Roche, is a highly selective BCL-2 inhibitor (Andrew, J.; Joel, D.). et al. (Nature Medicine, 2013, 19(2), 202) In the treatment of relapsed / refractory chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), and multiple myeloma (MM), the combination of Valentin and other drugs with Ibrutinib significantly improved the objective response rate (ORR) and complete response rate (CR) (Valentin, R.; Grablow, S.). et al. While BCL-2 inhibitors have been used (Blood, 2018, 132(12), 1248), they still have toxic side effects such as leukopenia and thrombocytopenia, anemia, diarrhea, dizziness, fatigue, and increased susceptibility to infection. Severe side effects include pneumonia, anemia, and high fever. Therefore, it is necessary to develop highly active and low-toxicity selective BCL-2 inhibitors. Summary of the Invention

[0005] This invention relates to compounds of formula (I), their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts. in: X 1 The substituent is selected from optionally substituted C3-C6 cycloalkyl groups or optionally substituted 3-6 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S, wherein the optional substituent is selected from hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, or 3-6 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S; preferably, X 1The substituent is selected from optionally substituted C5-C6 cycloalkyl groups or optionally substituted 5-6 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S, wherein the optional substituent is selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, or 4- or 5 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S; More preferably, X 1 The group is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxane, piperidinyl, morpholinyl, wherein the optional substituent is selected from hydroxyl, C1-C4 alkyl, oxetyl, or tetrahydrofuranyl; most preferably, X 1 Selected from (1) r 4 r )-1-hydroxy-1-methylcyclohexane-4-yl, ( S )-1,4-dioxane-2-yl, ( R )-1,4-dioxane-2-yl, tetrahydropyran-4-yl, 1-(oxetane-3-yl)piperidin-4-yl, ( S )-4-(oxetane-3-yl)morpholin-2-yl; X 2 Selected from 5-6 membered heterocyclic alkyl groups containing one or two heteroatoms selected from N, O, and S, wherein the heterocyclic alkyl group is optionally substituted with one or two C1-C4 alkyl groups or a halogen; preferably, X 2 Selected from 6-membered heterocyclic alkyl groups containing 2 nitrogen atoms, wherein the heterocyclic alkyl group is optionally substituted with 1 or 2 C1-C4 alkyl groups; more preferably, X 2 Selected from wherein the piperazine group is optionally substituted with one C1-C4 alkyl group; more preferably, X 2 Selected from wherein the piperazine group is optionally substituted with one methyl group; most preferably, X 2 Selected from , ; R 0 Selected from hydrogen and halogens; preferably, R 0 Selected from hydrogen, fluorine, and chlorine; most preferably, R 0 Selected from hydrogen and fluorine; R 1 R 2 Each is independently selected from hydrogen and C1-C6 alkyl groups; preferably, R 1 R 2 Each is independently selected from hydrogen and C1-C4 alkyl groups; more preferably, R 1 R 2 Each is independently selected from hydrogen, methyl, and ethyl; most preferably, R 1 R 2Each is independently selected from hydrogen and methyl; n is selected from 1 to 4; preferably, n is selected from 1, 3 or 4.

[0006] Preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts, wherein: X 1 Selected from optionally substituted C5-C6 cycloalkyl groups or optionally substituted 5-6 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S, wherein the optional substituents are selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, or 4- or 5 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, and S. X 2 Selected from 6-membered heterocyclic alkyl groups containing 2 N atoms, wherein the heterocyclic alkyl group is optionally substituted with 1 or 2 C1-C4 alkyl groups; R 0 Selected from hydrogen and halogens; R 1 R 2 Each is independently selected from hydrogen and C1-C6 alkyl groups; n is selected from 1-4.

[0007] More preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts, wherein: X 1 The substituted group is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxane, piperidinyl, morpholinyl, wherein the optional substituent is selected from hydroxyl, C1-C4 alkyl, oxetyl butyl or tetrahydrofuranyl. X 2 Selected from The piperazine group is optionally substituted with one C1-C4 alkyl group; R 0 Selected from hydrogen and halogens; R 1 R 2 Each is independently selected from hydrogen and C1-C4 alkyl groups; n is selected from 1-4.

[0008] More preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts, wherein: X 1The substituted group is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxane, piperidinyl, morpholinyl, wherein the optional substituent is selected from hydroxyl, C1-C4 alkyl, oxetyl butyl or tetrahydrofuranyl. X 2 Selected from The piperazine group is optionally substituted with one C1-C4 alkyl group; R 0 Selected from hydrogen, fluorine, and chlorine; R 1 R 2 Each is independently selected from hydrogen and C1-C4 alkyl groups; n is selected from 1, 3, or 4.

[0009] More preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts, wherein: X 1 Selected from (1) r 4 r )-1-hydroxy-1-methylcyclohexane-4-yl, ( S )-1,4-dioxane-2-yl, ( R )-1,4-dioxane-2-yl, tetrahydropyran-4-yl, 1-(oxetane-3-yl)piperidin-4-yl, ( S )-4-(oxetane-3-yl)morpholin-2-yl; X 2 Selected from , wherein the piperazine group is optionally substituted with one methyl group; R 0 Selected from hydrogen and fluorine; R 1 R 2 Each is independently selected from hydrogen, methyl, and ethyl; n is selected from 1, 3, or 4.

[0010] More preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts, wherein: X 1 Selected from (1) r 4 r )-1-hydroxy-1-methylcyclohexane-4-yl, ( S )-1,4-dioxane-2-yl, ( R )-1,4-dioxane-2-yl, tetrahydropyran-4-yl, 1-(oxetane-3-yl)piperidin-4-yl, ( S)-4-(oxetane-3-yl)morpholin-2-yl; X 2 Selected from , ; R 0 Selected from hydrogen and fluorine; R 1 R 2 Each is independently selected from hydrogen and methyl; n is selected from 1, 3, or 4.

[0011] More preferably, the present invention relates to compounds of formula (I) as described above, their isomers, prodrugs, solvates, stable isotopic derivatives or pharmaceutically acceptable salts thereof, selected from: .

[0012] The present invention also relates to the use of compounds of formula (I), isomers thereof, prodrugs, solvates, stable isotopic derivatives thereof or pharmaceutically acceptable salts thereof as described in any embodiment of the present invention in the preparation of medicaments used as BCL-2 inhibitors.

[0013] The present invention also relates to the use of compounds of formula (I) or isomers thereof, prodrugs, solvates, stable isotopic derivatives thereof or pharmaceutically acceptable salts thereof, according to any embodiment of the present invention, in the preparation of medicaments for the treatment or prevention of BCL-2-mediated diseases, such as tumors, selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0014] The present invention further relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer thereof as described in any embodiment of the present invention, a prodrug, a solvate, a stable isotopic derivative thereof or a pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients.

[0015] The present invention also relates to the use of the pharmaceutical composition according to the invention in the preparation of a medicament for treating or preventing BCL-2-mediated diseases, such as tumors, the tumors being selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0016] The present invention also relates to a method for treating or preventing BCL-2-mediated diseases, comprising administering to a patient in need a therapeutically effective amount of a compound or isomer thereof, prodrug, solvate, stable isotope derivative thereof, or pharmaceutically acceptable salt thereof, as described in any embodiment of the present invention; or a pharmaceutical composition thereof, wherein the related disease is, for example, a tumor selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0017] Another aspect of the present invention relates to compounds, isomers thereof, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts described in any embodiment of the present invention, for the treatment or prevention of BCL-2-mediated diseases, such as tumors selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0018] Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer thereof as described in any embodiment of the present invention, a prodrug, a solvate, a stable isotope derivative thereof or a pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients, for the treatment or prevention of BCL-2-mediated diseases, such as tumors selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0019] Another aspect of the invention relates to compounds of formula (I) or isomers thereof, prodrugs, solvates, stable isotopic derivatives thereof, or pharmaceutically acceptable salts thereof, as a treatment and / or prevention of BCL-2-mediated related diseases. The BCL-2-mediated related diseases include, for example, tumors selected from hematologic malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

[0020] According to the present invention, the drug can be any dosage form, including but not limited to tablets, capsules, solutions, lyophilized preparations, and injections.

[0021] The pharmaceutical formulations of the present invention can be administered in dose units containing a predetermined amount of the active ingredient per dose unit. Such units may contain, for example, 0.5 mg to 1 gram, preferably 1 mg to 700 mg, and particularly preferably 5 mg to 300 mg of the compound of the present invention, depending on the condition being treated, the method of administration, and the patient's age, weight, and condition. Alternatively, the pharmaceutical formulation can be administered in dose units containing a predetermined amount of the active ingredient per dose unit. Preferred dose unit formulations are those containing the daily dose or fractional dose, or a corresponding fraction thereof, as indicated above. Furthermore, this type of pharmaceutical formulation can be prepared using methods known in the pharmaceutical industry.

[0022] The pharmaceutical formulations of the present invention are suitable for administration by any desired and suitable method, such as oral (including oral cavity or sublingual), rectal, nasal, local (including oral cavity, sublingual, or percutaneous), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal) administration. Such formulations can be prepared using all methods known in the pharmaceutical field, for example, by combining the active ingredient with one or more excipients or one or more adjuvants.

[0023] Preparation process The present invention also provides a method for preparing the compound.

[0024] Process 1 R 0 With X 1 The definition is as described above; first step: Compound (I) was dissolved in chlorosulfonic acid and reacted in an oil bath at 120–150 °C for 10–20 hours. After cooling to room temperature, the reaction was quenched with ice water, extracted with ethyl acetate, and the crude product obtained by drying and concentrating the organic phase was dissolved in anhydrous tetrahydrofuran. Ammonia was added dropwise at low temperature (-80–-60 °C) and stirring was continued for 1–5 hours. The mixture was then acidified with an acid such as hydrochloric acid to obtain compound (II). Step Two: Compound (II) and the corresponding amine are dissolved in a solvent (such as acetonitrile), and a base (such as triethylamine or diisopropylethylamine) is added. The mixture is stirred for 10 to 20 hours under the protection of an inert gas (such as nitrogen or argon) at a temperature of 25 to 60 °C to obtain compound (III).

[0025] Process 2 R 1 R 2 The definitions of n and n are as described above; first step: Phosphorus oxychloride was added dropwise to an ice bath under nitrogen protection.N , N A dichloromethane solution of dimethylformamide was added dropwise, and the mixture was stirred at room temperature for 30 minutes. The temperature was then lowered to 0 °C, and a dichloromethane solution of compound (IV) was added dropwise. The mixture was reacted at room temperature to 60 °C for 10 to 24 hours to obtain compound (V). Step Two: Under nitrogen protection, compound (V), p-chlorophenylboronic acid, base such as potassium carbonate, phase transfer catalyst such as tetra-n-butylammonium bromide, and catalyst such as palladium acetate were added to a solvent such as water. The system was evacuated and replaced with nitrogen three times, and heated to 40-100 °C for 2-10 hours to obtain compound (VI). Step 3: Compound (VI) was dissolved in a solvent such as tetrahydrofuran or methanol, and a reducing agent such as sodium borohydride was added. The mixture was stirred at room temperature for 1 to 5 hours to obtain compound (VII). Step 4: Compound (VII) was dissolved in a solvent such as dichloromethane, and a chlorination reagent such as thionyl chloride was added. The mixture was stirred at room temperature for 10 to 24 hours to obtain compound (VIII).

[0026] Process 3 R 0 R 1 R 2 n and X 1 The definition is as stated above; R 3 Selected from H or methyl; first step: The compound (IX) (synthesis reference: Journal of Organic Chemistry , 84(8), 4814-4829,2019) and the corresponding piperazine or methylpiperazine (X) are dissolved in a solvent (such as dimethyl sulfoxide), and an alkali is added at room temperature. N , N -Diisopropylethylamine was stirred at 50–100 °C for 12–24 hours to obtain compound (XI); Step Two: Compounds (XI) and (VIII) are dissolved in a solvent such as acetonitrile, and an alkali such as... N , N -Diisopropylethylamine, heated to 50-90 °C and stirred for 8-24 hours, yields compound (XII); Step 3: Compound (XII) is dissolved in a solvent such as water or ethanol, a base such as lithium hydroxide is added, the mixture is heated to 50-90 °C and stirred for 1-5 hours, and then acidified with an acid such as hydrochloric acid to obtain compound (XIII). Step 4: Compound (XIII), compound (III), condensing agent such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and base such as 4-dimethylaminopyridine are dissolved in a solvent such as dichloromethane and stirred at room temperature for 12 to 36 hours to obtain compound (XIV). Detailed Implementation

[0027] definition Unless otherwise stated, the terms used in the specification and claims have the following meanings. Groups not specifically defined in this invention have meanings commonly understood in the art by those skilled in the art.

[0028] The notation “Cx-Cy” used in this invention represents the range of carbon atoms, where x and y are both integers. For example, C3-C8 cycloalkyl represents a cycloalkyl with 3-8 carbon atoms, and C0-C2 alkyl represents an alkyl with 0-2 carbon atoms, where C0 alkyl refers to a chemical single bond.

[0029] In this invention, the term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms, such as straight-chain and branched groups with 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, and various branched isomers thereof. The alkyl group may be optionally substituted or unsubstituted.

[0030] In this invention, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 12 ring atoms, for example, 3 to 12, 3 to 10, 3 to 8, or 3 to 6 ring atoms, or a 3, 4, 5, or 6-membered ring. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The cycloalkyl group may be optionally substituted or unsubstituted.

[0031] In this invention, the term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon group comprising 3 to 20 ring atoms, for example, 3 to 16, 3 to 12, 3 to 10, 3 to 8, or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O)m (where m is an integer from 0 to 2), but excluding ring portions of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon. Preferably, it comprises 3 to 12 ring atoms, wherein 1 to 4 are heteroatoms; more preferably, the heterocyclic group comprises 3 to 10 ring atoms; even more preferably, it comprises 3 to 8 ring atoms; most preferably, it is a 5-membered or 6-membered ring, wherein 1 to 4 are heteroatoms; more preferably, 1 to 3 are heteroatoms; and most preferably, 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include oxobutyl, pyrrolyl, piperidinyl, 4-piperidinyl, piperazine, 1,4-dioxane, morpholinyl, 2-morpholinyl, 4-morpholinyl, thiomorpholinyl, pyranyl, tetrahydropyranyl, 4-tetrahydropyranyl, homopiperazine, dioxyl, 2-dioxyl, tetrahydrofuranyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups. Heterocyclic groups can be optionally substituted or unsubstituted.

[0032] In this invention, the term "hybrid heterocyclic group" refers to a substituted or unsubstituted heterocyclic group having two terminal monovalent cores, which is produced by removing one hydrogen atom from each of the two terminal atoms; the heterocyclic group has the meaning described above. Non-limiting examples of "hybrid heterocyclic group" include pyrrolidinyl, piperidinyl, piperazineyl, morpholinyl, etc.

[0033] In this invention, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0034] In this invention, "optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or absence of the event or environment. For example, "optionally alkyl-substituted heterocyclic group" means that an alkyl group may but does not have to be present, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0035] In this invention, "substituted" refers to one or more hydrogen atoms in a group, preferably up to five, and more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0036] The substituents include, but are not limited to, the various groups described above.

[0037] The compounds claimed in this invention include not only the compound itself, but also isomers of the compound, prodrugs, solvates, stable isotopic derivatives or pharmaceutically acceptable salts thereof.

[0038] The term "pharmaceutical composition" as used in this invention refers to a mixture containing one or more isomers of the compounds described in this invention, prodrugs, solvates, stable isotopic derivatives, or pharmaceutically acceptable salts thereof, and other chemical components. Other components include, for example, pharmaceutically acceptable carriers, diluents, and excipients. The purpose of the pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0039] When used in the specification, the term "comprising" includes "consisting of".

[0040] The "room temperature" mentioned in this invention refers to 15-30℃.

[0041] The "stable isotope derivatives" of this invention include: isotope-substituted derivatives obtained by replacing any hydrogen atom in formula (I) with 1-5 deuterium atoms, isotope-substituted derivatives obtained by replacing any carbon atom in formula (I) with 1-3 carbon-14 atoms, or isotope-substituted derivatives obtained by replacing any oxygen atom in formula (I) with 1-3 oxygen-18 atoms.

[0042] The "pharmaceutically acceptable salt" described in this invention is found in Berge, et al. The discussion is in “Pharmaceutically Acceptable Salts”, J. Pharm. Sci., 66, 1-19 (1977), and it is obvious to medicinal chemists that the salts described are substantially non-toxic and provide the desired pharmacokinetic properties, palatability, absorption, distribution, metabolism, or excretion.

[0043] The pharmaceutically acceptable salts of this invention can be synthesized by conventional chemical methods.

[0044] Generally, salts can be prepared by reacting a free base or acid with an equistoichiometric or excess amount of an acid (inorganic or organic) or base in a suitable solvent or solvent combination.

[0045] The term "prodrug" as used in this invention refers to a compound that is metabolized in vivo and converted into its original active compound. Typically, a prodrug is an inactive substance, or one with less activity than the active parent compound, but it provides convenient handling, administration, or improved metabolic properties.

[0046] The term "isomer" in this invention refers to tautomers, mesosomes, racemates, enantiomers, diastereomers, and mixtures thereof of the compound of formula (I) of this invention. All such isomers, including stereoisomers and geometric isomers, are included in this invention. Geometric isomers include cis-trans isomers.

[0047] The term "solvent" as used in this invention refers to the association of one or more solvent molecules with a compound or salt thereof of this invention. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, ethyl acetate, acetic acid, etc.

[0048] This invention includes any polymorph of the compound or its salts, as well as any hydrates or other solvates.

[0049] In this invention, the term "patient" generally refers to mammals, especially humans.

[0050] In this invention, the term "tumor" includes both benign and malignant tumors, such as cancer.

[0051] In this invention, the term "cancer" includes various tumors mediated by BCL-2, including but not limited to hematologic malignancies such as acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, and glioma.

[0052] In this invention, the term "therapeutic effective amount" refers to the amount of the compound of this invention that can effectively treat or prevent related diseases mediated by BCL-2. Example

[0053] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0054] The structures of all compounds in this invention can be identified by nuclear magnetic resonance (¹H NMR) and / or mass spectrometry (MS).

[0055] 1 ¹H NMR chemical shifts (δ) were recorded in ppm (parts per million). NMR was performed using a Bruker AVANCE III 400 MHz spectrometer. Suitable solvents were selected from deuterated chloroform (CDCl₃), deuterated methanol (CD₃OD), and deuterated dimethyl sulfoxide (DMSO). d 6 Tetramethylsilane was used as an internal standard (TMS).

[0056] Low-resolution mass spectrometry (MS) was performed using an Agilent 1260 HPLC / 6120 mass spectrometer with an Agilent ZORBAXXDB-C18, 4.6 × 50 mm, 3.5 μm.

[0057] Gradient elution conditions 1: 0-1 min: 95% solvent A1 and 5% solvent B1; 1-2 min: 5% solvent A1 and 95% solvent B1; 2.01-2.50 min: 95% solvent A1 and 5% solvent B1. Percentages represent the volume percentage of a specific solvent in the total solvent volume. Solvent A1: 0.01% formic acid aqueous solution; Solvent B1: 0.01% formic acid in acetonitrile solution; percentages represent the volume percentage of the solute in the solution.

[0058] Thin-layer silica gel plates are generally selected from Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. Column chromatography generally uses Yantai Huanghai 100-200 or 200-300 mesh silica gel as the carrier.

[0059] Preparative liquid chromatography (prep-HPLC) was performed using a Waters SQD2 mass-guided high-performance liquid chromatograph, XBridge-C18; 30 x 150 mm preparative column, 5 μm. Method 1: Acetonitrile-water (0.2% formic acid), flow rate 25 mL / min; Method 2: Acetonitrile-water (0.8% ammonium bicarbonate), flow rate 25 mL / min; The known starting materials of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Shanghai Bid Pharmaceutical, Shanghai Aladdin Chemical, Shanghai Mairui Chemical, Bailingwei Chemical, and Anaiji Chemical.

[0060] Unless otherwise specified in the examples, all solvents used in the reactions were anhydrous. Commercially available tetrahydrofuran was used, with sodium carbonate as the dehydrating agent and benzophenone as the indicator. The mixture was refluxed under argon protection until the solution turned blue-purple, collected by distillation, and stored at room temperature under argon protection. Other anhydrous solvents were purchased from Anaiji Chemical and Bailingwei Chemical. All transfers and uses of anhydrous solvents, unless otherwise specified, must be carried out under argon protection.

[0061] Unless otherwise specified in the examples, the reactions were carried out under an argon or nitrogen atmosphere.

[0062] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1 L.

[0063] A hydrogen atmosphere refers to a reaction vessel connected to a hydrogen balloon with a volume of approximately 1 L.

[0064] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0065] Unless otherwise specified in the examples, the reaction temperature is room temperature, and the temperature range is 15℃ - 30℃.

[0066] In the examples, the reaction process was monitored using thin-layer chromatography (TLC). The developing solvent systems used were: A) dichloromethane and methanol; and B) petroleum ether and ethyl acetate. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0067] The eluent systems for column chromatography and the developing solvent systems for thin-layer chromatography used to purify the compounds include: A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents is adjusted according to the polarity of the compounds, and can also be adjusted by adding small amounts of triethylamine and acidic or basic reagents.

[0068] Intermediate 1 3-Fluoro-5-nitro-4-(((tetrahydro-2- H -pyran-4-yl)methyl)amino)benzenesulfonamide first step 3,4-Difluoro-5-nitrobenzenesulfonamide 1,2-Difluoro-3-nitrobenzene (10.00 g, 62.89 mmol) was dissolved in chlorosulfonic acid (21 mL) and heated to 150 °C under reflux with stirring for 10 hours. After cooling to room temperature, a saturated sodium bicarbonate solution was added to the reaction mixture in an ice bath to adjust the pH to approximately 7. Extraction was performed with dichloromethane (100 mL × 3), the organic phase was washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product 3,4-difluoro-5-nitrobenzenesulfonyl chloride. Isopropanol (200 mL) and ammonia (5 mL, 37%) were added to a 1000 mL three-necked flask and stirred at -78 °C for 10 minutes. The crude product, 3,4-difluoro-5-nitrobenzenesulfonyl chloride, was dissolved in isopropanol (30 mL) and slowly added dropwise to the above mixture of isopropanol and ammonia at -78 °C. After the addition was complete, the mixture was stirred at -78 °C for two hours. Dilute hydrochloric acid (1 N) was added to adjust the pH of the system to approximately 6. The reaction mixture was then brought to room temperature and concentrated under reduced pressure to remove most of the isopropanol solvent. Pure water was added, and a solid precipitated out. The solid crude product was filtered to obtain the crude product. The crude product was purified by slurrying with dichloromethane to obtain the target product, 3,4-difluoro-5-nitrobenzenesulfonamide (4.90 g, yellow solid). Yield: 33%.

[0069] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.38-8.36 (m, 1H), 8.29-8.26 (m, 1H), 7.84(s, 2H).

[0070] Step 2 3-Fluoro-5-nitro-4-(((tetrahydro-2- H -pyran-4-yl)methyl)amino)benzenesulfonamide Compound 3,4-difluoro-5-nitrobenzenesulfonamide (1.55 g, 6.51 mmol), (tetrahydro-2... H 4-pyran-4-yl)methylamine (0.89 g, 7.73 mmol), N , N - Diisopropylethylamine (3.91 g, 30.31 mmol) and acetonitrile (20.0 mL) were mixed. The mixture was stirred at 40 °C for 2 hours. The solvent was evaporated, and the mixture was quenched with 50 mL of water and extracted with ethyl acetate (60 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 3). The separated organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure to obtain the crude product. Column chromatography (petroleum ether / ethyl acetate = 1:1) was used to purify the target product 3-fluoro-5-nitro-4-(((tetrahydro-2-)- H 4-pyran-methyl)amino)benzenesulfonamide (1.66 g, yellow solid). Yield: 76%.

[0071] MS m / z (ESI): 334 [M + 1]; 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.33-8.30 (m, 2H), 7.76-7.74 (m, 1H), 7.45(s, 2H), 3.86-3.84 (m, 2H), 3.50-3.45 (m, 2H), 3.29-3.23 (m, 3H), 1.59-1.57(m, 2H), 1.25-1.20(m, 2H).

[0072] The synthesis steps for intermediate 2 are the same as those for intermediate 1, where the second step uses ( S )-1,4-dioxane-2-methylamine replaces (tetrahydro-2H-pyran-4-yl)methylamine.

[0073] Intermediate 3 3-Fluoro-5-nitro-4-(((1-(3-oxetane-butyl)piperidin-4-yl)methyl)amino)benzenesulfonamide first step 4-(((2-fluoro-6-nitro-4-aminosulfonylphenyl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester The compound 3,4-difluoro-5-nitrobenzenesulfonamide (0.30 g, 1.26 mmol) and tert-butyl 4-(aminomethyl)piperidine-1-carboxylate (0.28 g, 1.28 mmol) were used. N , N - Diisopropylethylamine (0.47 g, 3.63 mmol) and acetonitrile (8 mL) were mixed and stirred at 40 °C for 2 hours. The mixture was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was evaporated to dryness. The residue was purified by column chromatography (petroleum ether / acetic acid = 3:2) to give the target product tert-butyl 4-(((2-fluoro-6-nitro-4-aminosulfonylphenyl)amino)methyl)piperidine-1-carboxylate (0.49 g, yellow solid). Yield: 89%.

[0074] MS m / z (ESI): 433 [M + 1]; 1 H NMR (400 MHz, CD3OD) δ 8.41-8.37 (m, 1H), 7.64-7.60 (m, 1H), 4.02-4.00 (m, 2H), 3.47-3.44 (m, 2H), 2.67-2.64 (m, 2H), 1.75-1.66 (m, 3H), 1.35 (s, 9H), 1.14-1.03 (m, 2H).

[0075] Step 2 3-Fluoro-5-nitro-4-((piperidin-4-ylmethyl)amino)benzenesulfonamide Compound tert-butyl 4-(((2-fluoro-6-nitro-4-aminosulfonylphenyl)amino)methyl)piperidine-1-carboxylate (0.49 g, 1.12 mmol) was mixed with trifluoroacetic acid (3 mL) and dichloromethane (9 mL) and stirred at room temperature for 0.5 hours. The solvent was evaporated to dryness, and the mixture was adjusted to pH neutral with triethylamine and evaporated to dryness again to give crude 3-fluoro-5-nitro-4-((piperidine-4-ylmethyl)amino)benzenesulfonamide (0.35 g, yellow oil). This mixture was used directly in the next reaction without purification. MS m / z (ESI): 333 [M + 1]; Step 3 3-Fluoro-5-nitro-4-(((1-(3-oxetane-butyl)piperidin-4-yl)methyl)amino)benzenesulfonamide Compound 3-fluoro-5-nitro-4-(((piperidin-4-ylmethyl)amino)benzenesulfonamide (0.35 g, 1.05 mmol), oxetane-3-one (0.23 g, 3.15 mmol), sodium cyanoborohydride (0.23 g, 5.23 mmol), and methanol (8 mL) were mixed and stirred at room temperature for 1.5 hours. The mixture was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The organic phases were combined and washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and evaporated to dryness. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 3:1) to give the target product 3-fluoro-5-nitro-4-(((1-(3-oxetane-4-ylmethyl)amino)benzenesulfonamide (0.32 g, yellow solid). Yield: 78%.

[0076] MS m / z (ESI): 389 [M + 1]; 1 H NMR (400 MHz, CD3OD) δ 8.47-8.46 (m, 1H), 7.72-7.70 (m, 1H), 4.68-4.65 (m, 2H), 4.60-4.59 (m, 2H), 3.57-3.55 (m, 2H), 3.49-3.45 (m, 1H), 2.83-2.80 (m, 2H), 1.89-1.78 (m, 3H), 1.42-1.20 (m, 4H).

[0077] Intermediate 4 (3a R 7a S )-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H -Indigo first step (3a R 7a S )-6-chloro-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -Indene-5-formaldehyde Will N , N Dimethylformamide (8.76 g, 0.12 mol) and anhydrous dichloromethane (300 mL) were mixed and cooled to 0°C in an ice bath. Phosphorus oxychloride (13.86 g, 0.09 mol) was then slowly added dropwise. After the addition was complete, the mixture was stirred at 0°C for 30 minutes, then brought to room temperature and stirred for 3 hours. The mixture was then cooled to 0°C in an ice bath, and compound (3a) was slowly added dropwise. S 7a R )-7a-methyloctahydro-5 H A mixture of 9.00 g (0.06 mol) and anhydrous dichloromethane (50 mL) was added dropwise and then stirred at room temperature for 48 hours. The mixture was quenched with saturated ammonium chloride solution (300 mL), the organic phase was separated, dissolved under reduced pressure, and the residue was mixed with the aqueous phase and extracted with methyl tert-butyl ether (150 mL × 3). The combined organic phases were washed with saturated brine (150 mL × 2), dried with anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was dissolved under reduced pressure to obtain the target product ((3a)). R 7a S )-6-chloro-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H Indene-5-carboxaldehyde (10.30 g, yellow liquid), yield: 88%. MS m / z (ESI): 199 & 201 [M + 1]; Step 2 (3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -Indene-5-formaldehyde Compound ((3a) R 7a S )-6-chloro-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 HIndene-5-carboxaldehyde (10.30 g, 0.05 mol), 4-chlorophenylboronic acid (7.80 g, 0.05 mol), and water (300 mL) were mixed, and palladium acetate (1.13 g, 5.00 mmol), potassium carbonate (20.70 g, 0.15 mol), and tetrabutylammonium bromide (16.10 g, 0.05 mol) were added under argon protection. The mixture was stirred at 50 °C for 6 hours under argon protection. The mixture was quenched with water (100 mL) and methyl tert-butyl ether (200 mL). The organic phase was separated, and the aqueous phase was extracted with methyl tert-butyl ether (150 mL × 2). The combined organic phases were washed with saturated brine (150 mL × 2), dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was desolventized under reduced pressure to obtain the crude product, which was purified by rapid column chromatography (petroleum ether / ethyl acetate = 95:5) to give the target product (3a). R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H Indene-5-carboxaldehyde (4.00 g, yellow liquid). Yield: 28%.

[0078] MS m / z (ESI): 275 & 277 [M + 1]; 1 H NMR (400 MHz, CDCl3) δ 9.52 (s, 1H), 7.44-7.30 (m, 2H), 7.20-7.02(m, 2H), 2.72-2.33 (m, 3H), 2.11-1.74 (m, 4H), 1.72-1.22 (m, 4H), 1.02 (s,3H).

[0079] Step 3 ((3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -indene-5-yl)methanol Compound (3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 HIndene-5-carboxaldehyde (1.00 g, 3.65 mmol) and tetrahydrofuran (40 mL) were mixed, and sodium borohydride (0.28 g, 7.30 mmol) was added. The mixture was stirred at room temperature for 1 hour. This mixture was quenched with saturated ammonium chloride solution (60 mL) and dichloromethane (60 mL). The organic phase was separated, and the aqueous phase was extracted with dichloromethane (30 mL × 3). The combined organic phases were washed with saturated brine (80 mL × 2), dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was desolvated under reduced pressure to give the target product ((3a)). R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H 5-indenyl)methanol (0.95 g, pale yellow liquid). Yield: 94%. MS m / z (ESI): 259 & 261 [M - 17]; Step 4 (3a R 7a S )-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -Indigo Compound ((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H 3,44 mmol) of indene-5-yl)methanol (0.95 g), thionyl chloride (3.0 mL), and dichloromethane (30 mL) were mixed and stirred at room temperature for 1 hour. Desolventization under reduced pressure yielded the target product (3a). R 7a S )-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -Indene (1.01 g, pale yellow liquid). Crude product. MS m / z (ESI): 295, 297 & 299 [M + 1].

[0080] The synthesis of intermediate 5 follows the same steps as intermediate 4, wherein the first step uses (3a) R 7a S )-7a-methyloctahydro-5 H -Indene-5-one (synthetic reference: Tetrahedron Letters, 35(1), 171-174; 1994) replacing (3a) S 7a R )-7a-methyloctahydro-5 H 1-indole-5-one.

[0081] Intermediate 6 (1 R 6 S )-4-(chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene first step (1 S 6 R )-4-chloro-6-methylbicyclo[4.1.0]hept-3-ene-3-carboxaldehyde Dry N,N Dimethylformamide (4.66 g, 63.83 mmol) was added to dry dichloromethane (160 mL), and phosphorus oxychloride (9.76 g, 63.83 mmol) was slowly added dropwise at room temperature, stirring for 3 hours. (1) R , 6 R 1-Methylbicyclo[4.1.0]hepta-3-one (3.96 g, 31.92 mmol) (synthesis reference: Tetrahedron Letters, 60(11), 785-788; 2019) was dissolved in dry dichloromethane (80 mL) and slowly added dropwise to the reaction solution. The mixture was stirred overnight at room temperature. The reaction was detected by TLC. The reaction solution was then slowly added dropwise to ice-cold saturated ammonium chloride aqueous solution. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (200 mL × 2). The combined organic phases were washed with saturated brine (200 mL × 2), dried with anhydrous sodium sulfate, filtered, and the solvent was removed from the filtrate under reduced pressure to obtain the crude product (1 S 6 R 4-Chloro-6-methylbicyclo[4.1.0]hept-3-ene-3-carboxaldehyde (4.25 g, pale yellow oil). Yield: 28%.

[0082] 1 H NMR (400 MHz, CDCl3) δ 10.18 (s, 1H), 2.9-2.69 (m, 3H), 2.47-2.40 (m, 1H), 1.15 (s, 3H), 1.02-0.97 (m, 1H), 0.43-0.40 (m, 1H), 0.33-0.31 (m,1H).

[0083] Step 2 (1 R 6 S)-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-ene-3-carboxaldehyde (1) S 6 R 4-Chloro-6-methylbicyclo[4.1.0]hept-3-en-3-carboxaldehyde (0.45 g, 2.68 mmol), p-chlorophenylboronic acid (0.50 g, 3.20 mmol), and tetrabutylammonium bromide (1.30 g, 4.00 mmol) were dissolved in water (10 mL), and potassium carbonate (1.10 g, 8.00 mmol) and palladium acetate (0.06 g, 0.27 mmol) were added. The mixture was stirred at 45 °C for 3 hours under nitrogen protection. After cooling to room temperature, the mixture was extracted with ethyl acetate (30 mL × 3), and the combined organic phases were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to give the target product (1). R , 6 S )-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-ene-3-carboxaldehyde (0.23 g, yellow oil).

[0084] MS m / z (ESI): 247 & 249 [M + 1]; 1 H NMR (400 MHz, CDCl3) δ 9.47 (s, 1H), 7.35 (d, J = 8.0 Hz, 2H), 7.12(d, J = 8.0 Hz, 2H), 2.95-2.90 (m, 1H), 2.80-2.66 (m, 2H), 2.50-2.42 (m, 1H), 1.18 (s, 3H), 1.08-1.03 (m, 1H), 0.43-0.36 (m, 2H).

[0085] Step 3 ((1 S 6 R )-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-en-3-ylmethanol (1) R 6 S3.24 g (13.10 mmol) of 4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-ene-3-carboxaldehyde was added to tetrahydrofuran (50 mL) and methanol (10 mL), and sodium borohydride (0.71 g, 19.70 mmol) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour. The mixture was quenched with saturated ammonium chloride solution (100 mL), extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give the crude product ((1 S 6 R 4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-en-3-yl)methanol (3.43 g, yellow oil). MS m / z (ESI): 249 & 251 [M + 1].

[0086] Step 4 (1 R 6 S )-4-(chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene ((1) S 6 R )-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-en-3-yl)methanol (3.43 g, 13.10 mmol) was dissolved in dry dichloromethane (100 mL), and added N,N Dimethylformamide (48 mg, 0.66 mmol). Thionyl chloride (3.12 g, 26.2 mmol) was dissolved in dichloromethane (50 mL) and slowly added dropwise to the reaction solution under ice bath conditions. The mixture was stirred at room temperature for 3 hours. The reaction was confirmed by TLC on a silica gel plate. The solvent was removed under reduced pressure, and the crude product was dissolved in dichloromethane. [The remaining text appears to be incomplete and requires further context.] N,N -Diisopropylethylamine (5 mL), stirred for 10 minutes, concentrated by rotary evaporation to obtain crude product (1 mL). R 6 S )-4-(chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene (3.50 g, yellow oil).

[0087] 1 H NMR (400 MHz, CDCl3) δ 7.36 (d, J = 8.0 Hz, 2H), 7.20 (d, J= 8.0,2H), 3.86 (s, 2H), 2.62-2.60 (m, 1H), 2.56-2.49 (m, 2H), 2.42-2.40 (m, 1H),1.13 (s, 3H), 1.00-0.96 (m, 1H), 0.49-0.47 (m, 1H), 0.34-0.32 (m, 1H).

[0088] The synthesis of intermediate 7 follows the same steps as intermediate 6, wherein the first step uses (1 S 6 S )-1-methylbicyclo[4.1.0]hepta-3-one (synthetic reference: Tetrahedron Letters, 60(11), 785-788; 2019) replacing (1 R 6 R )-1-methylbicyclo[4.1.0]hepta-3-one.

[0089] Intermediate 8 (4a S ,8a R 6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a-octahydronaphthalene first step (4a S ,8a R )-4a-methyloctahydro-1 H -spiro[naphthalene-2,2'-[1,3]dioxolane] Compound (4a) S ,8a R )-4a-methylhexahydro-1 H -spiro[naphthalene-2,2'-[1,3]dioxolane]-5(3 H )-ketone (synthesis reference: Organic Letters, 20(1), 130-133; 2018) (3.36 g, 15.00 mmol), hydrazine hydrate (10.00 g, 0.18 mmol), and potassium hydroxide (8.40 g, 0.15 mmol) were added to diethylene glycol (120 mL), heated to 240 °C and stirred for 2 hours, then cooled to room temperature. The reaction solution was diluted with water (500 mL), and the mixture was extracted with ethyl acetate (150 mL × 3). The organic phases were combined and washed with water (200 mL) and saturated brine (200 mL), respectively. The organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain product (4a).S ,8a R )-4a-methyloctahydro-1 H -spiro[naphthalene-2,2'-[1,3]dioxolane] (2.90 g, colorless oil). Yield: 92%.

[0090] 1 H NMR (400 MHz, CDCl3) δ 3.95-3.91 (m, 4H), 1.75-1.62 (m, 5H), 1.53-1.45 (m, 5H), 1.44-1.33 (m, 5H), 0.99 (s, 3H).

[0091] Step 2 (4a S ,8a R )-4a-methyloctahydronaphthalene-2-(1 H )ketone Compound (4a) S ,8a R )-4a-methyloctahydro-1 H Spiro[naphthalene-2,2'-[1,3]dioxolane] (2.90 g, 13.80 mmol) was dissolved in tetrahydrofuran (20 mL), and concentrated hydrochloric acid (37%, 10 mL) was added. After stirring at room temperature for 1 hour, water (100 mL) was added for dilution, and the tetrahydrofuran was removed by concentration. The aqueous phase was extracted with methyl tert-butyl ether (50 mL × 3), and the organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give product (4a). S ,8a R )-4a-methyloctahydronaphthalene-2-(1 H ) ketone (2.00 g, colorless oil). Yield: 87.2%. MS m / z (ESI): 167 [M +1]; Step 3 (4a R ,8a S 3-Chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carboxaldehyde Phosphorus oxychloride (5.50 g, 36.00 mmol) was added dropwise to an ice bath under nitrogen protection. N , N The dimethylformamide (5.30 g, 72.00 mmol) was added dropwise to a dichloromethane solution (50 mL). After the addition was complete, the mixture was heated to room temperature and stirred for 0.5 hours. The mixture was then cooled to 0 °C, and compound (4a) was added dropwise. S ,8a R)-4a-methyloctahydronaphthalene-2-(1 H Ketone (2.00 g, 12.00 mmol) was stirred overnight at room temperature. 20 mL of 40% sodium acetate aqueous solution was added, and the mixture was stirred at room temperature for 0.5 hours. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with water (30 mL × 2) and saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give product (4a). R ,8a S 3-Chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carboxaldehyde (2.30 g, yellow oil). Yield: 90%. MS m / z (ESI): 213 & 215 [M + 1]; Step 4 (4a R ,8a S )-3-(4-chlorophenyl) - 8a-Methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carboxaldehyde (4a) R ,8a S 3-Chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carboxaldehyde (2.30 g, crude product, 10.80 mmol), p-chlorophenylboronic acid (2.00 g, 13.00 mmol), potassium carbonate (4.50 g, 32.40 mmol), tetrabutylammonium bromide (3.50 g, 10.80 mmol), and palladium acetate (0.49 g, 2.20 mmol) were added to water (60 mL). The system was evacuated and purged three times with nitrogen. The mixture was heated to 50 °C and stirred for 4 hours. After cooling to room temperature, the reaction mixture was extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL). The separated organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0-5% ethyl acetate / petroleum ether) to give (4a... R ,8a S )-3-(4-chlorophenyl) - 8a - methyl - 1,4,4a,5,6,7,8,8a - Octahydronaphthalene-2-carboxaldehyde (0.90 g, pale yellow oil). Yield: 33.1%.

[0092] MS m / z (ESI): 289 & 291 [M + 1]; 1H NMR (400 MHz, CDCl3) δ 9.43 (s, 1H), 7.39-7.32 (m, 2H), 7.18-7.11(m, 2H), 2.61-2.52 (m, 1H), 2.45-2.35 (m, 1H), 2.23-2.13 (m, 1H), 1.86-1.78(m, 1H), 1.78-1.65 (m, 1H), 1.50-1.37 (m, 3H), 1.33-1.04 (m, 5H), 0.89 (s,3H).

[0093] Step 5 (4a R ,8a S )-3-(4-chlorophenyl) - 8a - methyl - 1,4,4a,5,6,7,8,8a - Octahydronaphthalene-2-methanol (4a) R ,8a S )-3-(4-chlorophenyl) - 8a - methyl - 1,4,4a,5,6,7,8,8a - Octahydronaphthalene-2-carboxaldehyde (0.90 g, 3.10 mmol) was dissolved in a tetrahydrofuran / methanol mixture (v / v = 10 / 1, 11 mL), and sodium borohydride (0.24 g, 6.20 mmol) was added. The mixture was stirred at room temperature for 2 hours. The reaction was quenched by adding saturated ammonium chloride solution (30 mL). The reaction solution was extracted with ethyl acetate (30 mL × 2), and the combined organic phases were washed with saturated brine (30 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain (4a R ,8a S )-3-(4-chlorophenyl) - 8a - methyl - 1,4,4a,5,6,7,8,8a - Octahydronaphthalene-2-methanol (0.63 g, yellow oil). Yield: 69.5%. MS m / z (ESI): 273 & 275 [M - 17]; Step 6 (4a S ,8a R 6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a-octahydronaphthalene (4a)R ,8a S )-3-(4-chlorophenyl) - 8a - methyl - 1,4,4a,5,6,7,8,8a - Octahydronaphthalene-2-methanol (0.63 g, 2.17 mmol) was dissolved in dichloromethane (20 mL), and thionyl chloride (0.77 g, 6.51 mmol) was added. The mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain (4a) S ,8a R 6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a-octahydronaphthalene (0.67 g, yellow oil). Crude product.

[0094] 1 H NMR (400 MHz, CDCl3) δ7.34-7.28 (m, 2H), 7.17-7.11 (m, 2H), 4.00-3.84 (m, 2H), 2.36-2.26 (m, 2H), 2.09-1.98 (m, 2H), 1.86-1.80 (m, 1H), 1.73-1.65 (m, 2H), 1.49-1.30 (m, 6H), 0.97 (s, 3H).

[0095] The synthesis of intermediate 9 is similar to that of intermediate 8, wherein the first step uses (4a) R ,8a S )-4a-methylhexahydro-1 H -spiro[naphthalene-2,2'-[1,3]dioxolane]-5(3 H )-ketone (synthetic reference: Organic Letters, 20(1), 130-133;2018) replacing (4a S ,8a R )-4a-methylhexahydro-1 H -spiro[naphthalene-2,2'-[1,3]dioxolane]-5(3 H )-ketone.

[0096] Intermediate 11 5-(chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H -Indigo first step 6-Carbonyloctahydro-1 HMethyl indene-5-carboxylate Dimethyl carbonate (0.45 g, 5.00 mol) and tetrahydrofuran (10 mL) were mixed, and sodium hydride (0.08 g, 2.00 mol, dispersed in 60% mineral oil) was added in portions under argon protection. The mixture was stirred at 50 °C for 5 minutes. Then octahydro-5 H 6-Indene-5-one (synthesis reference: Advanced Synthesis & Catalysis, 360(20), 3924-3929; 2018) (0.14 g, 1.00 mol) was stirred at 70 °C for 2 hours. The mixture was quenched with water (25 mL) and ethyl acetate (25 mL), and the organic phase was separated. The aqueous phase was extracted with ethyl acetate (25 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was desolvated under reduced pressure to obtain the crude product. The crude product was purified by preparative thin-layer chromatography (petroleum ether / ethyl acetate = 30:1) to give the target product 6-carbonyloctahydro-1- H Methyl indene-5-carboxylate (0.07 g, yellow liquid). Yield: 37%.

[0097] 1 H NMR (400 MHz, CDCl3) δ 3.76 (s, 3H), 2.61-2.57 (m, 1H), 2.22-2.18(m, 2H), 2.02-1.97 (m, 2H), 1.76-1.70 (m, 3H), 1.45-1.40 (m, 3H), 1.18-1.12(m, 2H).

[0098] Step 2 6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7,7a-hexahydro-1 H Methyl indene-5-carboxylate The compound 6-carbonyloctahydro-1 H Methyl indene-5-carboxylate (0.07 g, 0.37 mmol), 1,1,1-trifluoro- N -phenyl- N -((trifluoromethyl)sulfonyl)methanesulfonamide (0.21 g, 0.6 mmol), potassium carbonate (0.14 g, 1.0 mmol), N , NDimethylformamide (3 mL) and tetrahydrofuran (3 mL) were mixed and stirred at 45 °C for 15 hours under argon protection. After cooling to room temperature, the mixture was quenched with water (25 mL) and ethyl acetate (25 mL), and the organic phase was separated. The aqueous phase was extracted with ethyl acetate (25 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was desolvated under reduced pressure to obtain the crude product. The crude product was purified by preparative thin-layer chromatography (petroleum ether / ethyl acetate = 30:1) to give the target product 6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7,7a-hexahydro-1 H Methyl indene-5-carboxylate (37 mg, yellow liquid). Yield: 57%. MS m / z (ESI): 329 [M + 1]; Step 3 6-(4-Chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H Methyl indene-5-carboxylate The compound (6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7,7a-hexahydro-1) H Methyl indene-5-carboxylate (37 mg, 0.20 mmol), 4-chlorophenylboronic acid (50 mg, 0.30 mmol), ethylene glycol dimethyl ether (4 mL), and methanol (2 mL) were mixed. Tetra(triphenylphosphine)palladium (20 mg, 0.02 mmol) and cesium fluoride (80 mg, 0.50 mmol) were added under argon protection, and the mixture was reacted at 70 °C for 15 hours under argon protection. The mixture was cooled to room temperature and quenched with water (25 mL) and ethyl acetate (25 mL). The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (25 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 2). The mixture was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was desolventized under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 30:1) to obtain the target product 6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H Methyl indene-5-carboxylate (40 mg, yellow liquid). Yield: 65%.

[0099] MS m / z (ESI): 291 & 293 [M + 1]; 1 H NMR (400 MHz, CDCl3) δ 7.26 (d, J = 8.0 Hz, 2H), 7.06 (d, J= 8.0Hz, 2H), 3.45 (s, 3H), 2.89-2.85 (m, 1H), 2.41-2.38 (m, 1H), 2.27-2.23 (m,1H), 2.16-2.12 (m, 1H), 2.00-1.96 (m, 2H), 1.73-1.68 (m, 3H), 1.47-1.39 (m, 3H).

[0100] Step 4 (6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H -indene-5-yl)methanol The compound 6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H Methyl indene-5-carboxylate (40 mg, 0.17 mmol) and tetrahydrofuran (3 mL) were mixed and cooled to -15 °C in an ice-salt bath. Lithium aluminum hydride (10 mg, 0.30 mmol) was added slowly in portions, and the mixture was allowed to rise naturally to room temperature for 3 hours. The mixture was then quenched with hydrochloric acid (1 M, 0.30 mmol, 0.3 mL). Extraction was performed with dichloromethane (25 mL × 3), and the combined organic phases were washed with saturated brine (25 mL × 3). The mixture was dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was dissolved under reduced pressure to obtain the target product (6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1- H 1,5-indene-5-yl)methanol (40 mg, pale yellow liquid), yield: 98%. MS m / z (ESI): 245 & 247 [M -17].

[0101] Step 5 5-(chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H -Indigo The compound (6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1) H 5-Indene-5-yl)methanol (40 mg, 0.13 mmol), thionyl chloride (0.1 mL), and dichloromethane (3 mL) were mixed and stirred at room temperature for 1 hour. Desolventization under reduced pressure yielded the target product 5-(chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1 H -Indene (40 mg, pale yellow liquid), crude product. MSm / z (ESI): 281, 283 & 285 [M + 1].

[0102] Intermediate 12 ( R)-2-((1 H methyl pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(3-methylpiperazin-1-yl)benzoate ( R )-2-((1 H methyl pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(3-methylpiperazin-1-yl)benzoate Compound 2-((1) H methyl pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-fluorobenzoate (synthesis reference: Journal of Organic Chemistry, 84(8), 4814-4829; 2019) (2.86 g, 10.00 mmol), ( R 2-Methylpiperazine (3.00 g, 30.00 mmol), N , N - Diisopropylethylamine (3.12 g, 24.18 mmol) and dimethyl sulfoxide (20 mL) were mixed. The mixture was stirred at 60 °C for 16 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (60 mL × 2). The combined organic phases were washed with saturated brine (50 mL × 3). The organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure to obtain the crude product. Column chromatography (dichloromethane / methanol = 90:10) was used to purify the target product (…). R )-2-((1 H Methyl pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(3-methylpiperazin-1-yl)benzoate (2.55 g, yellow solid). Yield: 70%.

[0103] MS m / z (ESI): 367 [M + 1]; 1 H NMR (400 MHz, CDCl3) δ 9.92 (s, 1H), 8.19 (d, J = 2.4 Hz, 1H), 7.91(d, J = 9.2 Hz, 1H), 7.52 (d, J = 2.4 Hz, 1H), 7.36-7.35 (m, 1H), 6.66-6.63(m, 1H), 6.43 (d, J = 1.6 Hz, 1H), 6.34 (d, J= 2.4 Hz, 1H), 3.79 (s, 3H), 3.54-3.48 (m, 2H), 3.05-3.02 (m, 1H), 2.93-2.84 (m, 2H), 2.77-2.71 (m, 1H), 2.42-2.37 (m, 1H), 1.08 (d, J = 6.4 Hz, 3H).

[0104] Example 1 2-((1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxo)-4-(4-(((3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -indene-5-yl)methyl)piperazin-1-yl)- N -((3-nitro-4-(((tetrahydro-2) H -pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide first step 2-((1 H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H Methyl inden-5-yl)methyl)piperazin-1-yl)benzoate Compound (3a) R 7a S )-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -Indene (intermediate 4) (1.01 g, 3.44 mmol), 2-((1 H methyl pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(piperazin-1-yl)benzoate (synthesis reference: WO2010138588 A2) (1.21 g, 3.44 mmol). N , N- Diisopropylethylamine (1.80 g, 13.76 mmol) and acetonitrile (30 mL) were mixed and stirred at 75 °C for 16 hours. After cooling to room temperature, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was desolventized under reduced pressure to obtain the crude product. The crude product was purified by rapid column chromatography (petroleum ether / ethyl acetate = 1:1) to give the target product 2-((1 H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H Methyl inden-5-yl)methyl)piperazin-1-yl)benzoate (0.95 g, white solid). Yield: 45%.

[0105] MS m / z (ESI): 611 & 613 [M + 1]; 1 H NMR (400 MHz, CDCl3) δ 9.74 (s, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.88(d, J = 8.9 Hz, 1H), 7.52 (d, J = 2.4 Hz, 1H), 7.39-7.33 (m, 1H), 7.25-7.21(m, 2H), 6.99-6.90 (m, 2H), 6.60-6.58 (m, 1H), 6.45-6.43 (m, 1H), 6.27 (s,1H), 3.78 (s, 3H), 3.24-3.02 (m, 4H), 2.90-2.65 (m, 2H), 2.45-1.88 (m, 6H), 1.83-1.50 (m, 6H), 1.46-1.19 (m, 3H), 1.00 (s, 3H).

[0106] Step 2 (2-((1 H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -inden-5-yl)methyl)piperazin-1-yl)benzoic acid 2-((1) H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H Methyl inden-5-yl)methyl)piperazin-1-yl)benzoate (0.95 g, 1.56 mmol), sodium hydroxide (0.30 g, 7.50 mmol), ethanol (20 mL), and water (1 mL) were mixed and stirred at 80 °C for 1 hour. The solvent was removed under reduced pressure, and the residue was added to water (30 mL) and hydrochloric acid (1 M, 15 mL). Extraction was performed with dichloromethane (30 mL × 3), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The solvent was removed under reduced pressure to give the target product (2-((1)). H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3a) R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H 1,5-indene-5-yl)methyl)piperazin-1-yl)benzoic acid (0.90 g, white solid), yield: 97%. MS m / z (ESI): 597 & 599 [M + 1]; Step 3 2-((1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxo)-4-(4-(((3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -indene-5-yl)methyl)piperazin-1-yl)- N -((3-nitro-4-(((tetrahydro-2) H -pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (2-((1) H -pyrrolo[2,3- b ]pyridin-5-yl)oxo)-4-(4-(((3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H 1-Indene-5-yl)methyl)piperazin-1-yl)benzoic acid (0.50 g, 1.00 mmol), 3-nitro-4-(((tetrahydro-2-yl)benzoic acidH 2-((1-pyran-4-yl)methyl)amino)benzenesulfonamide (0.44 g, 1.40 mmol) (synthesis reference: WO 2018041248 A1), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.38 g, 2.00 mmol), 4-dimethylaminopyridine (0.12 g, 1.00 mmol), triethylamine (0.20 g, 2.00 mmol), and dichloromethane (60 mL) were mixed and stirred at room temperature for 16 hours. The mixture was diluted with water (50 mL), the organic phase was separated, and the aqueous phase was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was dissolved under reduced pressure to obtain the crude product. The residue was purified by rapid column chromatography (dichloromethane / methanol = 97:3) to give the target product 2-((1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxo)-4-(4-(((3a R 7a S )-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1 H -indene-5-yl)methyl)piperazin-1-yl)- N -((3-nitro-4-(((tetrahydro-2) H -pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide 1 (0.27 g, pale yellow solid), yield: 30%.

[0107] MS m / z (ESI): 894 & 896 [M + 1]; 1H NMR (400 MHz, CDCl3) δ 10.35-9.99 (m, 2H), 8.89 (s, 1H), 8.52 (s,1H), 8.31-8.11 (m, 2H), 8.00-7.86 (m, 1H), 7.71 (s, 1H), 7.50 (s, 1H), 7.31-7.14 (m, 2H), 6.99-6.78 (m, 3H), 6.64-6.39 (m, 2H), 5.99 (s, 1H), 4.19-3.91(m, 2H), 3.53-3.37 (m, 2H), 3.33-3.21 (m, 2H), 3.16-3.01 (m, 4H), 2.96-2.65(m, 2H), 2.47-2.13 (m, 5H), 2.10-1.87 (m, 3H), 1.83-1.69 (m, 4H), 1.67-1.49(m, 3H), 1.46-1.21 (m, 5H), 0.98 (s, 3H).

[0108] The synthesis steps of Examples 2 to 31 are the same as those of Example 1.

[0109] Biological experiments Tests of BCL-2 bioactivity inhibition The effect of the compounds of this invention on the bioactivity of BCL-2 was evaluated using fluorescence polarization assays. The experimental methods are summarized as follows: The effect of compounds on BCL-2 bioactivity was assessed using an affinity assay based on fluorescence polarization principles, by detecting the influence of compounds on the binding activity of BCL-2 and leukemia pro-apoptotic protein (BIM). The reaction buffer contained the following components: PBS (pH 7.4, 3 mM Na2HPO4, 155 mM NaCl, 1 mM KH2PO4), 1 mM DTT; recombinant human Bcl-2 protein (catalog number 10195-H08E) was purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd., and diluted to 5 nM with the reaction buffer; FITC-labeled BIM peptide was purchased from Nanjing Genscript Biotech Co., Ltd., and diluted to 5 nM with the reaction buffer.

[0110] The compound was dissolved and diluted in 100% DMSO to 0.1, 1, and 10 μM, and then serially diluted 4-fold with DMSO to the lowest concentrations of 0.0061, 0.061, and 0.61 nM. Each concentration point was then diluted 50-fold with reaction buffer.

[0111] Add 3 μL of the compound solution and 12 μL of BCL-2 solution to a black 384-well plate, mix thoroughly, and incubate at room temperature for 15 minutes. Then add 15 μL of FITC-BIM solution, incubate the reaction mixture at room temperature in the dark for 30 minutes, and immediately detect fluorescence polarization on an Envision microplate reader (Perkin Elmer). The excitation wavelength was 480 nm, and the emission wavelength was 535 nm. In this experiment, the group without BCL-2 protein served as a negative control (100% inhibition), and the group with BCL-2 protein but without the compound served as a positive control (0% inhibition). The percentage of inhibition of BCL-2 affinity by the compound can be calculated using the following formula: Compound IC 50 The values ​​were calculated using the following formula from eight concentration points with XLfit (ID Business Solutions Ltd., UK) software: Y = Bottom+(Top-Bottom) / (1+10^((logIC 50 -X)*slope factor)) Where Y is the inhibition percentage, X is the logarithm of the concentration of the analyte, Bottom is the maximum inhibition percentage, Top is the minimum inhibition percentage, and slope factor is the curve slope coefficient.

[0112] Tests of BCL-XL bioactivity inhibition The effect of the compounds of this invention on the bioactivity of BCL-XL was evaluated using fluorescence polarization assays. The experimental methods are summarized as follows: An affinity assay based on fluorescence polarization was used to assess the effect of compounds on the binding activity of BCL-XL and BIM, thereby evaluating the effect of compounds on the bioactivity of BCL-XL. The reaction buffer contained the following components: PBS (pH 7.4, 3 mM Na2HPO4, 155 mM NaCl, 1 mM KH2PO4), 1 mM DTT; human recombinant Bcl-XL protein (catalog number 10455-H08E) was purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd., and diluted to 10 nM with the reaction buffer; FITC-labeled BIM peptide was purchased from Nanjing Genscript Biotech Co., Ltd., and diluted to 10 nM with the reaction buffer.

[0113] The compound was dissolved and diluted to 1 μM in 100% DMSO, and then serially diluted 4-fold with DMSO to the lowest concentration of 0.061 nM. Each concentration point was then diluted 50-fold with reaction buffer.

[0114] Add 3 μL of the compound solution and 12 μL of BCL-XL solution to a black 384-well plate, mix thoroughly, and incubate at room temperature for 15 minutes. Then add 15 μL of FITC-BIM solution, incubate the reaction mixture at room temperature in the dark for 30 minutes, and immediately detect fluorescence polarization on an Envision microplate reader (Perkin Elmer). The excitation wavelength was 480 nm, and the emission wavelength was 535 nm. In this experiment, the group without BCL-XL protein served as a negative control (100% inhibition), and the group with BCL-XL protein but without the compound served as a positive control (0% inhibition). The percentage of inhibition of BCL-XL affinity by the compound can be calculated using the following formula: Compound IC 50 The values ​​were calculated using the following formula from eight concentration points with XLfit (ID Business Solutions Ltd., UK) software: Y = Bottom+(Top-Bottom) / (1+10^((logIC 50 -X)*slope factor)) Where Y is the inhibition percentage, X is the logarithm of the concentration of the analyte, Bottom is the maximum inhibition percentage, Top is the minimum inhibition percentage, and slope factor is the curve slope coefficient.

[0115] RS4;11 cells (acute lymphoblastic leukemia cells) IC50 50 Measurement The effect of the compounds of the present invention on the proliferation of RS4;11 cells was evaluated using a luminescence cell viability assay.

[0116] The experimental methods are summarized as follows: The CellTilter-Glo (CTG) assay kit was used to detect the key indicator of ATP metabolism in living cells by employing a unique, highly sensitive and stable luciferase. The luminescent signal generated in the experiment was directly proportional to the number of viable cells in the culture medium, thereby detecting the cell proliferation status of RS4;11.

[0117] CellTilter-Glo reagent (Promega, G7572) consists of CellTilter-Glo lyophilized powder and CellTilter-Glo buffer. To use, simply dissolve the lyophilized powder in the buffer.

[0118] RS4;11 cells (ATCC, CRL-1873) were cultured in RPMI 1640 complete medium (Thermofisher, 72400-047) containing 10% FBS (GBICO, 10099-141) and 100 units / ml penicillin-streptomycin mixture (Thermofisher, 15140122). When the cell coverage in the culture vessel reached 80-90%, the cells were digested with 0.25% trypsin (containing EDTA) (Thermofisher, catalog number 25200056), dispersed, and seeded into white 384-well plates (Thermofisher, catalog number 164610). The 384-well plates were then incubated overnight at 37°C in a 5% CO2 incubator. The compound was dissolved and diluted to 5 mM in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.061 μM. Each concentration point was then diluted 50-fold using FBS-free RPMI 1640 medium. If the compound IC50... 50 The value is very low, allowing for a reduction in the initial concentration of the compound. After overnight incubation, 3 μL of the diluted compound in culture medium was added to each well, and the mixture was gently centrifuged to mix. The group without cells served as a negative control (100% inhibition), while the group with 0.2% DMSO served as a positive control (0% inhibition). The 384-well plate was incubated at 37°C with 5% CO2 for 48 hours. After equilibration to room temperature, 15 μL of CTG reagent was added to each well, and the plate was gently shaken on a shaker for 3 minutes to ensure complete cell lysis. The plate was then left to stand for 10 minutes to allow the cryo-light signal to stabilize, and the cryo-light signal was read using an EnVision (Perkin Elmer) scanner.

[0119] The percentage of inhibition of RS4;11 cell proliferation by the compound can be calculated using the following formula: Inhibition percentage = 100 - 100 * (signal compound - negative control) / (signal positive control - negative control) Compound IC 50 The values ​​were calculated from eight concentration points using the XLfit (ID Business Solutions Ltd., UK) software via the following formula: Y = Bottom + (Top- Bottom) / (1+10^((LogIC 50 -X) * slope factor)) Where Y represents the inhibition percentage, Bottom is the bottom plateau of the curve, Top is the top plateau of the curve, and X is the logarithm of the concentration of the analyte.

[0120] The results of the above in vitro BCL-2 and BCL-XL protein activity assays are shown in Table 1 below, and the results of the cell experiments are shown in Table 2.

[0121] Table 1: Results of BCL-2 and BCL-XL protein activity assays Compound numbering <![CDATA[FP BCL-2 IC 50 (nM)]]> <![CDATA[FP BCL-XL IC 50 (nM)]]> 1 6.14 >1000 2 9.13 >1000 3 2.62 >1000 4 4.01 >1000 5 2.46 >1000 6 3.53 >1000 7 1.7 >1000 8 12.15 >1000 9 8.25 >1000 10 14.51 >1000 11 7.37 >1000 12 7.14 >1000 13 3.28 >1000 14 7.03 >1000 15 3.49 >1000 16 2.9 >1000 17 5.39 >1000 18 17.68 >1000 19 9.22 >1000 20 15.04 >1000 21 19.53 >1000 22 7.14 >1000 23 2.11 >1000 24 4.99 >1000 25 10.13 >1000 26 0.58 >1000 27 0.85 >1000 28 3.99 >1000 29 2.89 >1000 30 1.5 >1000 31 1.2 >1000 .

[0122] Table 2: Results of RS4;11 cell viability assay Compound numbering <![CDATA[RS4;11 IC 50 (nM)]]> 1 19.5 3 9.0 5 4.75 6 10.77 7 2.8 8 83 9 29 10 30 11 41 12 37 13 12 14 19 15 37 16 11 17 15 23 14 24 16 25 37 26 4.5 27 16 28 5.8 29 6 30 26 31 13 Venetoclax 5.5 .

[0123] The experimental results above show that the compounds in the embodiments of the present invention can effectively and selectively inhibit the activity of BCL-2, with weaker inhibition of BCL-XL. They can be used to treat various cancers caused by abnormal overexpression of BCL-2 family proteins, especially hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma. Furthermore, they avoid the toxic side effects caused by BCL-XL inhibition, such as thrombocytopenia. Some compounds can also effectively inhibit the proliferation of RS4;11 acute lymphoblastic cells. They have a strong inhibitory effect on hematological malignancies such as acute lymphoblastic leukemia.

[0124] It will be apparent to those skilled in the art that this disclosure is not limited to the illustrative embodiments described above, and that it may be embodied in other specific forms without departing from the essential characteristics of this disclosure. Therefore, it is intended that these embodiments be considered illustrative and non-limiting in all respects, and that reference be made to the appended claims rather than the foregoing embodiments, and that all variations therein within the equivalent meaning and scope of the claims are included therein.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: 。 2. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament used as a BCL-2 inhibitor.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of BCL-2-mediated related diseases, wherein the related diseases are selected from the group consisting of: acute lymphoblastic leukemia, malignant hematologic diseases, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and glioma.

4. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable excipients.

5. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers and / or diluents.

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