WO2022148439A1 - Heterocyclic compound as bcl-2 inhibitor - Google Patents

Abstract

Provided in the present invention are a heterocyclic compound as a BCL-2 inhibitor as represented by formula (I), a pharmaceutical composition containing the heterocyclic compound, and the use of the heterocyclic compound for treating or preventing related diseases and dysfunction mediated by BCL-2, such as tumors. The present invention also relates to a method for preparing the heterocyclic compound.

Description

Heterocyclic compounds as BCL-2 inhibitors technical field

The present invention relates to heterocyclic compounds, pharmaceutical compositions containing them and their use as B-cell lymphoma-2 (BCL-2) inhibitors. More specifically, the present invention provides novel heterocyclic compounds as BCL-2 inhibitors, pharmaceutical compositions containing such heterocyclic compounds, and application of the heterocyclic compounds to treat or prevent BCL-2-mediated related diseases and functional disorders, such as tumors. The present invention also relates to a method for preparing the heterocyclic compound.

Background technique

The BCL-2 protein family is one of the core regulatory mechanisms of apoptosis (also called programmed cell death), which can receive and transmit internal intracellular signals or external environmental stress signals, such as nutritional or hypoxic stress, DNA damage, carcinogenesis Excessive activation of genes, endoplasmic reticulum stress, etc., mainly play a leading role in the intrinsic apoptosis pathway. 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 3 subclasses according to function and sequence analysis. The first subclass is anti-apoptotic, including BCL-XL, BCL-2, BCL-W, MCL -1, BFL-1, they are mainly localized on mitochondria and protect mitochondria from adversity damage; the other two subclasses are apoptosis-promoting, and one subclass is the ultimate executor of mitochondrial damage, including BAX and BAK. The rest belong to the BH3 subclass, which can directly sense various cellular stress signals. A dynamic balance of interactions between proteins that antagonize and promote apoptosis to determine the life and death fate of cells. BCL-2 protein, which antagonizes apoptosis, is closely related to tumors. About 50% of tumors (such as leukemia, rectal cancer, prostate cancer, etc.) have abnormal overexpression of BCL-2 family proteins. widely present in. Multiple signaling pathways, such as JAK-STAT, NFkB, and UPS (ubiquitin-proteasome system), can cause the overexpression of BCL-2 protein that antagonizes apoptosis.

The high expression of BCL-2 family anti-apoptotic proteins is related to the drug resistance of various tumors. For example, the overexpression of BCL-2 anti-apoptotic proteins can enable tumor cells to escape apoptosis caused by anti-tumor drugs, thereby causing drug resistance. medicine. Studies have shown that inhibition of BCL-2 family proteins can inhibit the formation of tumor angiogenesis, thereby inhibiting tumor metastasis (Benjamin, D.; Isaac, J. et al. J. Clin. Oncol. 2008, 26(25), 4180 ). Therefore, targeted inhibition of BCL-2 family anti-apoptotic proteins can inhibit the occurrence, development and drug resistance of tumors.

Although more than 20 small-molecule inhibitors targeting the BCL-2 family have been reported, very few have entered clinical trials, with Teva's Obatoclax being obtained in only 1 out of 26 treated chronic lymphocytic leukemia (CLL) patients. Partial response, with strong neurotoxicity, discontinued development in 2013; AbbVie's Navitoclax (ABT-263), despite its Phase I dose escalation in patients with relapsed or refractory lymphoid malignancies, showed 50% good response rate, but also showed very strong targeted toxicity of BCL-XL: 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), multiple myeloma (MM), etc., through combination therapy with Ibrutinib, etc., the objective response rate (ORR) and The complete remission rate (CR) has been greatly improved (Valentin, R.; Grablow, S. et al. Blood, 2018, 132(12), 1248), but there are still leukopenia and thrombocytopenia, anemia, diarrhea, dizziness , fatigue, susceptibility to infection and other toxic side effects, serious toxic side effects also include pneumonia, anemia, high fever, etc. Therefore, it is necessary to develop selective BCL-2 inhibitors with high activity and low toxicity.

SUMMARY OF THE INVENTION

The present invention relates to compounds of formula (I), isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof,

in:

X ¹ is selected from optionally substituted C3-C6 cycloalkyl or optionally substituted 3-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein said any The selected substituent is selected from hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S; preferably , X ¹ is selected from optionally substituted C5-C6 cycloalkyl or optionally substituted 5-6-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein the Optional substituents are selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, 4- or 5-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S; further preferably , X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, and morpholinyl, wherein the optional Substituents are selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl; most preferably, X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4- base, (S)-1,4-dioxan-2-yl, (R)-1,4-dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxa Cyclobutan-3-yl)piperidin-4-yl, (S)-4-(oxetan-3-yl)morpholin-2-yl;

其中所述亚哌嗪基任选被1个C1-C4烷基取代；再进一步优选地，X ²选自

其中所述亚哌嗪基任选被1个甲基取代；最优选地，X ²选自

X ² is selected from 5-6 membered heterocycloalkylene containing 1 or 2 heteroatoms selected from N, O, S, wherein the heterocycloalkylene is optionally surrounded by 1 or 2 C1-C4 alkyl groups or halogen substitution; preferably, X ² is selected from a 6-membered heterocycloalkylene group containing 2 N atoms, wherein the heterocycloalkylene group is optionally substituted with 1 or 2 C1-C4 alkyl groups; further preferably , X ² is selected from

wherein the piperazinide group is optionally substituted by 1 C1-C4 alkyl group; further preferably, X ² is selected from

wherein the piperazinylene group is optionally substituted with ¹ methyl; most preferably, X is selected from

R ⁰ is selected from hydrogen, halogen; preferably, R ⁰ is selected from hydrogen, fluorine, chlorine; most preferably, R ⁰ is selected from hydrogen, fluorine;

R ¹ and R ² are each independently selected from hydrogen and C1-C6 alkyl; preferably, R ¹ and R ² are each independently selected from hydrogen and C1-C4 alkyl; more preferably, R ¹ and R ² are each independently is selected from hydrogen, methyl, ethyl; most preferably, R ¹ , R ² are each independently selected from hydrogen, methyl;

n is selected from 1-4; preferably, n is selected from 1, 3 or 4.

Preferably, the present invention relates to compounds of formula (I) as hereinbefore described, isomers, prodrugs, solvates, stable isotopic derivatives or pharmaceutically acceptable salts thereof, wherein:

X ¹ is selected from optionally substituted C5-C6 cycloalkyl or optionally substituted 5-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein any of said The selected substituent is selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, 4- or 5-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S;

X ² is selected from a 6-membered heterocycloalkylene group containing 2 N atoms, wherein the heterocycloalkylene group is optionally substituted with 1 or 2 C1-C4 alkyl groups;

R ⁰ is selected from hydrogen, halogen;

R ¹ and R ² are each independently selected from hydrogen, C1-C6 alkyl;

n is selected from 1-4.

Further preferably, the present invention relates to compounds of formula (I) as hereinbefore described, isomers, prodrugs, solvates, stable isotopic derivatives or pharmaceutically acceptable salts thereof, wherein:

X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, morpholinyl, wherein the optionally substituted group is selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl;

其中所述亚哌嗪基任选被1个C1-C4烷基取代； X ² is selected from

wherein the piperazinylene group is optionally substituted by 1 C1-C4 alkyl group;

R ⁰ is selected from hydrogen, halogen;

R ¹ and R ² are each independently selected from hydrogen, C1-C4 alkyl;

n is selected from 1-4.

Still further preferably, the present invention relates to a compound of formula (I) as hereinbefore described, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein:

X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, morpholinyl, wherein the optionally substituted group is selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl;

其中所述亚哌嗪基任选被1个C1-C4烷基取代； X ² is selected from

wherein the piperazinylene group is optionally substituted by 1 C1-C4 alkyl group;

R ⁰ is selected from hydrogen, fluorine, chlorine;

R ¹ and R ² are each independently selected from hydrogen, C1-C4 alkyl;

n is selected from 1, 3 or 4.

More preferably, the present invention relates to a compound of formula (I) as hereinbefore described, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein:

X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4-yl, (S)-1,4-dioxan-2-yl, (R)-1,4 -Dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxetan-3-yl)piperidin-4-yl, (S)-4-(oxetane alk-3-yl)morpholin-2-yl;

其中所述亚哌嗪基任选被1个甲基取代； X ² is selected from

wherein the piperazinylene group is optionally substituted with 1 methyl;

R ⁰ is selected from hydrogen, fluorine;

R ¹ and R ² are each independently selected from hydrogen, methyl, and ethyl;

n is selected from 1, 3 or 4.

Still further preferably, the present invention relates to a compound of formula (I) as hereinbefore described, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein:

X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4-yl, (S)-1,4-dioxan-2-yl, (R)-1,4 -Dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxetan-3-yl)piperidin-4-yl, (S)-4-(oxetane alk-3-yl)morpholin-2-yl;

X ² is selected from

R ⁰ is selected from hydrogen, fluorine;

R ¹ and R ² are each independently selected from hydrogen and methyl;

n is selected from 1, 3 or 4.

Still further preferably, the present invention relates to a compound of formula (I) as described above, its isomers, prodrugs, solvates, stable isotopic derivatives or pharmaceutically acceptable salts thereof, selected from the group consisting of:

The present invention also relates to compounds of formula (I), isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof according to any one of the embodiments of the present invention for use in the preparation of Use of a BCL-2 inhibitor in medicine.

The present invention also relates to a compound of formula (I) or an isomer, prodrug, solvate, stable isotope derivative or a pharmaceutically acceptable salt thereof according to any one of the embodiments of the present invention in the manufacture of a medicament for the treatment or prevention of related diseases mediated by BCL-2, such as tumors selected from hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer , rectal cancer, prostate cancer, pancreatic cancer, glioma.

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

The present invention also relates to the use of the pharmaceutical composition according to the present invention in the preparation of a medicament, wherein the medicament is used for the treatment or prevention of BCL-2-mediated related diseases, such as tumors, which are selected from From hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, glioma.

The present invention also relates to a method of treating or preventing related diseases mediated by BCL-2, comprising administering to a patient in need thereof a therapeutically effective amount of the compound described in any one of the embodiments of the present invention or an isomer, pro- Drugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof; or pharmaceutical compositions of the present invention, said related diseases such as tumors selected from hematological malignancies including acute lymphoblastic leukemia disease, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, glioma.

Another aspect of the present invention pertains to a compound described in any one of the embodiments of the present invention, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, for use in therapy or Prevention of related diseases mediated by BCL-2, such as tumors selected from hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, Glioma.

Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer, prodrug, solvate, stable isotope thereof, as described in any one of the embodiments of the present invention Derivatives or pharmaceutically acceptable salts thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients for use in therapy Or prevent related diseases mediated by BCL-2, such as tumors selected from hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer ,Glioma.

Another aspect of the present invention pertains to a compound of formula (I) or an isomer, prodrug, solvate thereof as described in any one of the embodiments of the present invention for the treatment and/or prophylaxis of BCL-2-mediated related diseases , a stable isotope derivative or a pharmaceutically acceptable salt thereof. Said BCL-2 mediated related disease such as tumor, said related disease such as tumor selected from hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreas Cancer, glioma.

According to the present invention, the drug can be in any pharmaceutical dosage form, including but not limited to tablets, capsules, solutions, freeze-dried preparations, and injections.

The pharmaceutical formulations of the present invention may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit. Such a unit may contain, for example, from 0.5 mg to 1 gram, preferably from 1 mg to 700 mg, particularly preferably from 5 mg to 300 mg, of a compound of the invention, or drug, depending on the condition to be treated, the method of administration and the age, weight and condition of the patient. The formulations can be administered in dosage unit form containing a predetermined quantity of active ingredient per dosage unit. Preferred dosage unit formulations are those containing a daily dose or sub-dose, as indicated above, or a corresponding fraction thereof, of the active ingredient. Furthermore, pharmaceutical formulations of this type can be prepared using methods well known in the pharmaceutical art.

The pharmaceutical formulations of the present invention may be suitable for administration by any desired suitable method, such as oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods of administration. Such formulations can be prepared, for example, by combining the active ingredient with one or more excipients or one or more adjuvants, using all methods known in the art of pharmacy.

Preparation process

The present invention also provides methods for preparing the compounds.

Process 1

The definitions of R ⁰ and X ¹ are as described above;

first step:

Compound (I) was dissolved in chlorosulfonic acid, heated in an oil bath (120-150° C.) and reacted for 10-20 hours, cooled to room temperature, quenched with ice water, extracted with ethyl acetate, and the organic phase was dried and concentrated to obtain the crude product Dissolve in anhydrous tetrahydrofuran, add ammonia water dropwise at low temperature (-80～-60°C), continue stirring for 1～5 hours, and acidify with acid such as hydrochloric acid to obtain compound (II);

Step 2:

Compound (II) and the corresponding amine are dissolved in a solvent (such as acetonitrile), a base (such as triethylamine or diisopropylethylamine, etc.) is added, and under the protection of an inert gas (such as nitrogen or argon), at temperature The mixture was stirred at 25 to 60° C. for 10 to 20 hours to obtain compound (III).

Process 2

R ¹ , R ² and n are as defined above;

first step:

Under nitrogen protection, phosphorus oxychloride was added dropwise to the dichloromethane solution of N,N-dimethylformamide under an ice bath. After the dropwise addition, the mixture was stirred at room temperature for 30 minutes, cooled to 0°C, and compound (IV) was added dropwise. , react at room temperature to 60 °C for 10 to 24 hours to obtain compound (V);

Step 2:

Under nitrogen protection, compound (V), p-chlorobenzeneboronic acid, alkali such as potassium carbonate, phase-transfer catalyst such as tetra-n-butylammonium bromide and catalyst such as palladium acetate are added to a solvent such as water, the system is evacuated and replaced with nitrogen three times, and heated to 40 ℃. React at ~100°C for 2 to 10 hours to obtain compound (VI);

third step:

Dissolve compound (VI) in a solvent such as tetrahydrofuran or methanol, add a reducing agent such as sodium borohydride, and stir at room temperature for 1-5 hours to obtain compound (VII);

the fourth step:

Compound (VII) is dissolved in a solvent such as dichloromethane, a chlorinating reagent such as thionyl chloride is added, and the mixture is stirred at room temperature for 10-24 hours to obtain compound (VIII).

Process 3

R ⁰ , R ¹ , R ² , n and X ¹ are as defined above; R ³ is selected from H or methyl;

first step:

Compound (IX) (synthetic 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) , adding a base such as N,N-diisopropylethylamine at room temperature, stirring at a temperature of 50-100° C. for 12-24 hours to obtain compound (XI);

Step 2:

Compound (XI) and compound (VIII) are dissolved in a solvent such as acetonitrile, a base such as N,N-diisopropylethylamine is added, heated to 50-90° C. and stirred for 8-24 hours to obtain compound (XII);

third step:

Dissolving compound (XII) in a solvent such as water and ethanol, adding a base such as lithium hydroxide, heating to 50-90° C. and stirring for 1-5 hours, and then acidifying with an acid such as hydrochloric acid to obtain compound (XIII);

the fourth step:

Compound (XIII), compound (III), a condensing agent such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, a base such as 4-dimethylaminopyridine are dissolved in a solvent such as In dichloromethane, the mixture is stirred at room temperature for 12 to 36 hours to obtain compound (XIV).

Detailed ways

definition

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings. Groups not specifically defined in the present invention have the meanings generally represented in the art known to those skilled in the art.

The notation "Cx-Cy" used in the present invention represents a range of carbon atoms, where x and y are both integers, for example C3-C8 cycloalkyl represents a cycloalkyl having 3-8 carbon atoms, C0- C2 alkyl refers to an alkyl group having 0-2 carbon atoms, wherein C0 alkyl refers to a chemical single bond.

In the present invention, the term "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 20 carbon atoms, for example, may be 1 to 18 carbon atoms, 1 to 12 carbon atoms , straight and branched chain groups of 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, etc. Alkyl groups can be optionally substituted or unsubstituted.

In the present invention, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 12 ring atoms, such as 3 to 12, 3 to 10, 3 to 8 or 3 to 6 ring atoms, or can be a 3, 4, 5, or 6 membered ring. Non-limiting examples of monocyclic cyclyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Cycloalkyl groups can be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 20 ring atoms, such as 3 to 16, 3 to 12, 3 to 16 10, 3 to 8, or 3 to 6 ring atoms, one or more of which is selected from nitrogen, oxygen, or a heteroatom of S(O)m (where m is an integer from 0 to 2), excluding- The ring moiety of O-O-, -O-S- or -S-S-, the remaining ring atoms being carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably the heterocyclyl ring contains 3 to 10 ring atoms, more preferably 3 to 8 ring atoms, most preferably a 5- or 6-membered ring A ring of which 1-4 are heteroatoms, more preferably 1-3 are heteroatoms, and most preferably 1-2 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include oxetanyl, pyrrolidinyl, piperidinyl, 4-piperidinyl, piperazinyl, 1,4-dioxanyl, morpholinyl, 2-morpholinyl, 4-morpholinyl, thiomorpholinyl, pyranyl, tetrahydropyranyl, 4-tetrahydropyranyl, homopiperazinyl, dioxanyl, 2-dioxanyl Alkyl, tetrahydrofuranyl, etc. Polycyclic heterocyclyls include spiro, fused and bridged heterocyclyls. Heterocyclyl groups can be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclylene" refers to a substituted or unsubstituted heterocyclyl having a core of two terminal monovalent groups obtained by removing one hydrogen atom from each of the two terminal atoms. produced; the heterocyclyl group has the meanings set forth above. Non-limiting examples of "heterocyclylene" include pyrrolidylene, piperidinylene, piperazinylene, morpholinylene, and the like.

In the present invention, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group .

In the present invention, "substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of each other, are substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the person skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.

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

The compounds claimed in the present invention include not only the compounds themselves, but also isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof.

The "pharmaceutical composition" of the present invention refers to containing one or more isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof and other chemical components of the compounds of the present invention mixture. Other components such as pharmaceutically acceptable pharmaceutically acceptable carriers, diluents and excipients. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.

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

The "room temperature" in the present invention refers to 15-30°C.

The "stable isotopic derivatives" of the present invention include: isotopically substituted derivatives obtained by replacing any hydrogen atom in formula (I) with 1-5 deuterium atoms, and any carbon atom in formula (I) by 1-3 Isotopically substituted derivatives obtained by substituting one carbon 14 atom or isotopically substituted derivatives obtained by substituting 1-3 oxygen 18 atoms in the formula (I) for any oxygen atom.

The "pharmaceutically acceptable salts" of the present invention are discussed in Berge, et al., "Pharmaceutically acceptable salts", J. Pharm. It will be apparent that the salts are substantially non-toxic and provide the desired pharmacokinetic properties, palatability, absorption, distribution, metabolism or excretion and the like.

The pharmaceutically acceptable salts of the present invention can be synthesized by general chemical methods.

In general, salts can be prepared by reacting the free base or acid with an equivalent or an excess of acid (inorganic or organic) or base in a suitable solvent or solvent composition.

The "prodrug" in the present invention refers to the conversion of the compound into the original active compound after metabolism in the body. Typically, a prodrug is an inactive substance, or is less active than the active parent compound, but can provide ease of manipulation, administration, or improved metabolic properties.

The "isomers" in the present invention refer to the tautomers, mesomers, racemates, enantiomers, diastereomers, and other isomers of the compound of formula (I) of the present invention. Mixture form, etc. All of these isomers, including stereoisomers and geometric isomers, are included in the present invention. The geometric isomers include cis-trans isomers.

"Solvate" as used herein refers to the association of one or more solvent molecules with a compound of the present invention or a salt thereof. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, ethyl acetate, acetic acid, and the like.

The present invention includes any polymorphs of the compounds or salts thereof, as well as any hydrates or other solvates.

In the present invention, the term "patient" generally refers to mammals, especially humans.

In the present invention, the term "tumor" includes benign tumors and malignant tumors, such as cancer.

In the present invention, the term "cancer" includes various tumors mediated by BCL-2, including but not limited to hematological malignancies including acute lymphoblastic leukemia, lung, breast, ovarian, prostate, rectal, pancreatic Cancer, glioma.

In the present invention, the term "therapeutically effective amount" is meant to include an amount of the compound of the present invention effective to treat or prevent related diseases mediated by BCL-2.

Example

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the described examples. The experimental methods that do not specify specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product description.

The structures of all compounds of the present invention can be identified by nuclear magnetic resonance (1H NMR) and/or mass spectrometry (MS).

¹ H NMR chemical shifts (δ) are reported in PPM (parts per million). NMR was performed on a Bruker AVANCE III-400MHz spectrometer. Suitable solvents are selected from deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), deuterated dimethyl sulfoxide (DMSO-d ⁶ ), etc., with tetramethylsilane as internal standard (TMS).

Low resolution mass spectra (MS) were determined by an Agilent 1260 HPLC/6120 mass spectrometer using an Agilent ZORBAX XDB-C18, 4.6 x 50 mm, 3.5 μm.

Gradient elution condition one: 0-1 minutes: 95% solvent A1 and 5% solvent B1, 1-2 minutes: 5% solvent A1 and 95% solvent B1; 2.01-2.50 minutes: 95% solvent A1 and 5% solvent B1 . Percentage is the volume percent of a solvent in the total solvent volume. Solvent A1: 0.01% formic acid aqueous solution; solvent B1: 0.01% formic acid in acetonitrile solution; the percentage is the volume percentage of the solute in the solution.

Thin-layer silica gel plate is generally selected from Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. Column chromatography generally uses Yantai Yellow Sea 100-200 or 200-300 mesh silica gel as a carrier.

Preparative liquid chromatography (prep-HPLC) uses Waters SQD2 mass spectrometry-oriented high-pressure liquid chromatography, XBridge-C18; 30 × 150mm preparative column, 5 μm;

Method 1: acetonitrile-water (0.2% formic acid), flow rate 25mL/min; Method 2: acetonitrile-water (0.8% ammonium bicarbonate), flow rate 25mL/min;

The known starting materials of the present invention can be synthesized by adopting or according to methods known in the art, or can be purchased from Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Shanghai Bide Medicine, Shanghai Accela Latin Chemical, Shanghai Merrill Chemical, Bailingwei Chemical, Annagy Chemical and other companies.

Unless otherwise specified in the examples, the solvents used in the reaction are all anhydrous solvents. The anhydrous tetrahydrofuran was obtained by using commercially available tetrahydrofuran, using sodium block as water-removing agent and benzophenone as indicator, refluxed to a blue-purple solution under argon protection, collected by distillation, and stored at room temperature under argon protection. Other anhydrous solvents were purchased from Annagy Chemical and Bailingwei Chemical. All the transfer and use of anhydrous solvents should be carried out under the protection of argon unless otherwise specified.

Unless otherwise specified in the examples, the reactions were all carried out in an argon atmosphere or a nitrogen atmosphere.

Argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

Hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1 L.

The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.

Unless otherwise specified in the examples, the reaction temperature is room temperature, and the temperature range is 15°C-30°C.

The monitoring of the reaction progress in the examples adopts thin layer chromatography (TLC), and the systems of the developing solvents used in the reaction are A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound.

The eluent system for column chromatography and the developing solvent system for thin layer chromatography used to purify the compound include A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and can also be adjusted by adding a small amount of triethylamine and an acidic or basic reagent.

Intermediate 1

3-Fluoro-5-nitro-4-(((tetrahydro-2H-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), heated to 150° C. and stirred under reflux for 10 hours. After cooling to room temperature, a saturated aqueous sodium bicarbonate solution was added to the reaction solution under an ice bath to adjust the pH to about 7. Extract with dichloromethane (100 mL×3), wash the organic phase with saturated brine (100 mL×2), dry over anhydrous sodium sulfate, filter and concentrate to obtain the crude product 3,4-difluoro-5-nitrobenzenesulfonyl chloride. In a 1000 mL three-neck flask, isopropanol (200 mL) and ammonia water (5 mL, 37%) were added and stirred at -78°C for 10 minutes. The obtained crude product, 3,4-difluoro-5-nitrobenzenesulfonyl chloride, was dissolved in isopropanol (30 mL), and was slowly added dropwise to the above mixture of isopropanol and ammonia at -78°C. Stir at -78°C for two hours, and add dilute hydrochloric acid (1N) to adjust the pH of the system to about 6. Then the reaction mixture was warmed to room temperature, concentrated under reduced pressure to remove most of the isopropanol solvent, pure water was added thereto, the solid was precipitated, and a crude solid product was obtained by filtration. The crude product was purified by beating with dichloromethane to obtain the target product 3,4-difluoro-5-nitrobenzenesulfonamide (4.90 g, yellow solid). Yield: 33%.

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).

second step

3-Fluoro-5-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide

Compound 3,4-difluoro-5-nitrobenzenesulfonamide (1.55g, 6.51mmol), (tetrahydro-2H-pyran-4-yl)methanamine (0.89g, 7.73mmol), N,N - Diisopropylethylamine (3.91 g, 30.31 mmol) and acetonitrile (20.0 mL) were mixed. Stir at 40°C for 2 hours. The solvent was spun dry, the mixture was quenched with 50 mL of water, and extracted with ethyl acetate (60 mL x 2). The combined organic phases were washed with saturated brine (50 mL×3). The organic phase was separated and dried with anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolvation under reduced pressure. Purification by column chromatography (petroleum ether/ethyl acetate=1:1) to obtain the target product 3-fluoro-5-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino) Benzenesulfonamide (1.66 g, yellow solid). Yield: 76%.

MS m/z(ESI): 334[M+1];

¹ H NMR (400MHz, 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).

The synthetic steps of intermediate 2 refer to intermediate 1, wherein (S)-1,4-dioxane-2-methylamine is used instead of (tetrahydro-2H-pyran-4-yl)methanamine in the second step .

Intermediate 3

3-Fluoro-5-nitro-4-(((1-(3-oxetanyl)piperidin-4-yl)methyl)amino)benzenesulfonamide

first step

4-(((2-Fluoro-6-nitro-4-sulfamoylphenyl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester

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), 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 x 2). The combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, the filtrate was spin-dried, and the residue was purified by column chromatography (petroleum ether/acetic acid=3:2) to obtain the target product 4-(((2-fluoro-6- Nitro-4-sulfamoylphenyl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester (0.49 g, yellow solid). Yield: 89%.

MS m/z(ESI): 433[M+1];

¹ H NMR (400 MHz, CD ₃ OD) 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).

second step

3-Fluoro-5-nitro-4-((piperidin-4-ylmethyl)amino)benzenesulfonamide

Compound 4-(((2-fluoro-6-nitro-4-sulfamoylphenyl)amino)methyl)piperidine-1-carboxylic acid tert-butyl ester (0.49 g, 1.12 mmol) and trifluoroacetic acid ( 3 mL) and dichloromethane (9 mL), and stirred at room temperature for 0.5 hour. The solvent was spin-dried, the mixture was adjusted to neutral pH with triethylamine, and the solvent was spin-dried again to obtain crude 3-fluoro-5-nitro-4-((piperidin-4-ylmethyl)amino)benzenesulfonic acid Amide (0.35 g, yellow oil). This mixture was used directly in the next reaction without purification. MS m/z(ESI): 333[M+1];

third step

3-Fluoro-5-nitro-4-(((1-(3-oxetanyl)piperidin-4-yl)methyl)amino)benzenesulfonamide

Compound 3-fluoro-5-nitro-4-((piperidin-4-ylmethyl)amino)benzenesulfonamide (0.35 g, 1.05 mmol) and oxetan-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 x 2). The organic phases were combined and washed with saturated brine (10 mL). The organic phase was dried with anhydrous sodium sulfate, filtered to remove the desiccant, spin-dried, and the residue was purified by column chromatography (petroleum ether/ethyl acetate=3:1) to obtain the target product 3-fluoro-5-nitro- 4-(((1-(3-oxetanyl)piperidin-4-yl)methyl)amino)benzenesulfonamide (0.32 g, yellow solid). Yield: 78%.

MS m/z(ESI): 389[M+1];

¹ H NMR (400MHz, CD ₃ OD) 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).

Intermediate 4

(3aR,7aS)-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene

first step

(3aR,7aS)-6-Chloro-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carbaldehyde

N,N-dimethylformamide (8.76g, 0.12mol) and anhydrous dichloromethane (300mL) were mixed, the ice bath was lowered to 0°C, and phosphorus oxychloride (13.86g, 0.09mol) was slowly added dropwise. After the dropwise addition, the mixture was stirred at 0 °C for 30 minutes, then returned to room temperature and stirred for 3 hours. The ice bath was lowered to 0 °C, and the compound (3aS, 7aR)-7a-methyloctahydro-5H-inden-5-one was slowly added dropwise. (Synthesis reference: Tetrahedron Letters, 35(1), 171-174; 1994) (9.00 g, 0.06 mol) and anhydrous dichloromethane (50 mL) were added dropwise and returned to room temperature and stirred for 48 hours. The mixture was quenched with saturated ammonium chloride solution (300 mL), the organic phase was separated, desolvated under reduced pressure, 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 over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was desolvated under reduced pressure to obtain the target product ((3aR,7aS)-6-chloro-3a-methyl- 2,3,3a,4,7,7a-hexahydro-1H-indene-5-carbaldehyde (10.30 g, yellow liquid), yield: 88%. MS m/z (ESI): 199&201 [M+1] ;

second step

(3aR,7aS)-6-(4-Chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carbaldehyde

Compound ((3aR,7aS)-6-chloro-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carbaldehyde (10.30 g, 0.05 mol), 4- Chlorophenylboronic acid (7.80g, 0.05mol) and water (300mL) were mixed, and palladium acetate (1.13g, 5.00mmol), potassium carbonate (20.70g, 0.15mol) and tetrabutylammonium bromide ( 16.10 g, 0.05 mol), stirred at 50°C for 6 hours under argon. 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 washed with methyl tert-butyl ether. (150mL × 2) extraction. The combined organic phase was washed with saturated brine (150 mL × 2), dried with anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was removed under reduced pressure to obtain the crude product, which was purified by flash column chromatography (petroleum ether/ethyl acetate=95:5) to obtain the target product (3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro- 1H-Indene-5-carbaldehyde (4.00 g, yellow liquid). Yield: 28%.

MS m/z(ESI): 275&277[M+1];

¹ H NMR (400MHz, CDCl ₃ ) 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).

third step

((3aR,7aS)-6-(4-Chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methanol

The compound (3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carbaldehyde (1.00 g, 3.65 mmol) and tetrahydrofuran (40 mL) were mixed, sodium borohydride (0.28 g, 7.30 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The 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 removed under reduced pressure to obtain the target product ((3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene-5 - base) methanol (0.95 g, pale yellow liquid). Yield: 94%. MS m/z(ESI): 259&261[M-17];

the fourth step

(3aR,7aS)-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene

Compound ((3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methanol (0.95 g, 3.44 mmol), thionyl chloride (3.0 mL) and dichloromethane (30 mL) were mixed and stirred at room temperature for 1 hour. Precipitate under reduced pressure to obtain the target product (3aR,7aS)-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro -1H-Indene (1.01 g, pale yellow liquid). Crude. MS m/z (ESI): 295, 297 & 299 [M+1].

The synthesis of intermediate 5 refers to the synthesis steps of intermediate 4, wherein the first step uses (3aR, 7aS)-7a-methyloctahydro-5H-inden-5-one (synthetic reference: Tetrahedron Letters, 35(1), 171-174; 1994) in place of (3aS,7aR)-7a-methyloctahydro-5H-inden-5-one.

Intermediate 6

(1R,6S)-4-(chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene

first step

(1S,6R)-4-Chloro-6-methylbicyclo[4.1.0]hept-3-ene-3-carbaldehyde

Dry N,N-dimethylformamide (4.66 g, 63.83 mmol) was added to dry dichloromethane (160 mL), phosphorous oxychloride (9.76 g, 63.83 mmol) was slowly added dropwise at room temperature, and stirred for 3 hours . (1R,6R)-1-methylbicyclo[4.1.0]heptan-3-one (3.96 g, 31.92 mmol) (synthetic reference: Tetrahedron Letters, 60(11), 785-788; 2019) was dissolved in dry Dichloromethane (80 mL) was slowly added dropwise to the reaction solution and stirred at room temperature overnight. TLC spot plate detection reaction was completed, the reaction solution was slowly added dropwise to ice-cold saturated aqueous ammonium chloride solution, the organic phase was separated, the aqueous phase was extracted with dichloromethane (200 mL×2), and the organic phases were combined with saturated brine (200 mL). ×2) Wash, dry the organic phase with anhydrous sodium sulfate, filter, and remove the solvent from the filtrate under reduced pressure to obtain the crude product (1S,6R)-4-chloro-6-methylbicyclo[4.1.0]hept-3-ene- 3-Carboxaldehyde (4.25 g, pale yellow oil). Yield: 28%.

¹ H NMR (400MHz, CDCl ₃ ) 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).

second step

(1R,6S)-4-(4-Chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-ene-3-carbaldehyde

(1S,6R)-4-chloro-6-methylbicyclo[4.1.0]hept-3-ene-3-carbaldehyde (0.45 g, 2.68 mmol), p-chlorobenzeneboronic acid (0.50 g, 3.20 mmol), Tetrabutylammonium bromide (1.30 g, 4.00 mmol) was dissolved in water (10 mL), potassium carbonate (1.10 g, 8.00 mmol) and palladium acetate (0.06 g, 0.27 mmol) were added. Under nitrogen protection, it was stirred at 45°C for 3 hours. It was cooled to room temperature, extracted with ethyl acetate (30 mL×3), 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 obtain the target product (1R,6S)-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]heptane -3-ene-3-carbaldehyde (0.23 g, yellow oil).

MS m/z(ESI): 247&249[M+1];

¹ H NMR (400 MHz, CDCl ₃ ) 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).

third step

((1S,6R)-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-en-3-ylmethanol

(1R,6S)-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-ene-3-carbaldehyde (3.24 g, 13.10 mmol) was added to tetrahydrofuran (50 mL) and To methanol (10 mL), sodium borohydride (0.71 g, 19.70 mmol) was added portionwise at 0°C. Stir at room temperature for 1 hour. Saturated ammonium chloride solution (100 mL) was added to quench, extracted with ethyl acetate (50 mL×3), the combined organic phases were washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product ((1S,6R)-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].

the fourth step

(1R,6S)-4-(chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene

((1S,6R)-4-(4-chlorophenyl)-6-methylbicyclo[4.1.0]hept-3-en-3-yl)methanol (3.43 g, 13.10 mmol) was dissolved in dry Chloromethane (100 mL) was added N,N-dimethylformamide (48 mg, 0.66 mmol). Thionyl chloride (3.12 g, 26.2 mmol) was dissolved in dichloromethane (50 mL), slowly added dropwise to the reaction solution under an ice bath, and stirred at room temperature for 3 hours. The completion of the reaction was detected by TLC silica gel plate, the solvent was removed under reduced pressure, the obtained crude product was dissolved in dichloromethane, N,N-diisopropylethylamine (5 mL) was added, stirred for 10 minutes, and concentrated by rotary evaporation to obtain the crude product (1R, 6S) -4-(Chloromethyl)-3-(4-chlorophenyl)-1-methylbicyclo[4.1.0]hept-3-ene (3.50 g, yellow oil).

¹ H NMR (400 MHz, CDCl ₃ ) 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) ).

The synthesis of intermediate 7 refers to the synthesis steps of intermediate 6, wherein the first step uses (1S,6S)-1-methylbicyclo[4.1.0]heptan-3-one (synthetic reference: Tetrahedron Letters, 60(11) , 785-788; 2019) in place of (1R,6R)-1-methylbicyclo[4.1.0]heptan-3-one.

Intermediate 8

(4aS,8aR)-6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a-octahydronaphthalene

first step

(4aS,8aR)-4a-Methyloctahydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]

The compound (4aS,8aR)-4a-methylhexahydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-5(3H)-one (synthetic reference: Organic Letters, 20(1), 130-133; 2018) (3.36g, 15.00mmol), hydrazine hydrate (10.00g, 0.18mmol), potassium hydroxide (8.40g, 0.15mmol), diethylene glycol (120mL) was added, and the temperature was raised to After stirring at 240°C for 2 hours, it was cooled to room temperature. The reaction solution was diluted with water (500 mL), 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 phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was Concentration under reduced pressure gave the product (4aS,8aR)-4a-methyloctahydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (2.90 g, colorless oil). Yield: 92%.

¹ H NMR (400MHz, CDCl ₃ ) 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).

second step

(4aS,8aR)-4a-Methyloctahydronaphthalene-2-(1H)one

Compound (4aS,8aR)-4a-methyloctahydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (2.90 g, 13.80 mmol) was dissolved in tetrahydrofuran (20 mL), Concentrated hydrochloric acid (37%, 10 mL) was added, stirred at room temperature for 1 hour, diluted with water (100 mL), and concentrated to remove tetrahydrofuran. The aqueous phase was extracted with methyl tert-butyl ether (50 mL×3), 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 obtain the product (4aS, 8aR) -4a-Methyloctahydronaphthalen-2-(1H)one (2.00 g, colorless oil). Yield: 87.2%. MS m/z(ESI): 167[M+1];

third step

(4aR,8aS)-3-Chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carbaldehyde

Phosphorus oxychloride (5.50 g, 36.00 mmol) was added dropwise to a dichloromethane solution (50 mL) of N,N-dimethylformamide (5.30 g, 72.00 mmol) in an ice bath under nitrogen protection. , warmed to room temperature and stirred for 0.5 h. It was cooled to 0° C. again, compound (4aS, 8aR)-4a-methyloctahydronaphthalene-2-(1H)one (2.00 g, 12.00 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. A 40% aqueous sodium acetate solution (20 mL) was added, and the mixture was stirred at room temperature for 0.5 hour. The organic phase was separated, 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 , the product (4aR,8aS)-3-chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carbaldehyde (2.30 g, yellow oil) was obtained. Yield: 90%. MS m/z(ESI): 213&215[M+1];

the fourth step

(4aR,8aS)-3-(4-Chlorophenyl)-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carbaldehyde

(4aR,8aS)-3-chloro-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carbaldehyde (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), tetra-n-butylammonium 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 replaced with nitrogen three times, and the temperature was raised to 50°C and stirred for 4 hours. Cooled to room temperature, the reaction solution 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, the filtrate was concentrated under reduced pressure, and the residue was washed with Silica gel column separation and purification (0-5% ethyl acetate/petroleum ether) gave (4aR,8aS)-3-(4-chlorophenyl)-8a-methyl-1,4,4a,5,6,7, 8,8a-Octahydronaphthalene-2-carbaldehyde (0.90 g, pale yellow oil). Yield: 33.1%.

MS m/z(ESI): 289&291[M+1];

¹ H NMR (400MHz, CDCl ₃ ) 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).

the fifth step

(4aR,8aS)-3-(4-Chlorophenyl)-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-methanol

(4aR,8aS)-3-(4-chlorophenyl)-8a-methyl-1,4,4a,5,6,7,8,8a-octahydronaphthalene-2-carbaldehyde (0.90 g, 3.10 mmol) was dissolved in a tetrahydrofuran/methanol mixed solution (v/v=10/1, 11 mL), sodium borohydride (0.24 g, 6.20 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched by the addition of saturated ammonium chloride solution (30 mL). The reaction solution was extracted with ethyl acetate (30 mL×2), the combined organic phases were washed with saturated brine (30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain (4aR,8aS)-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

(4aS,8aR)-6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a-octahydronaphthalene

(4aR,8aS)-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), thionyl chloride (0.77 g, 6.51 mmol) was added and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain (4aS,8aR)-6-(chloromethyl)-7-(4-chlorophenyl)-4a-methyl-1,2,3,4,4a,5,8,8a - Octahydronaphthalene (0.67 g, yellow oil). Crude.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.34-7.28 (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).

The synthesis of intermediate 9 refers to the synthesis of intermediate 8, in which the first step uses (4aR,8aS)-4a-methylhexahydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane ]-5(3H)-one (Synthetic reference: Organic Letters, 20(1), 130-133; 2018) instead of (4aS,8aR)-4a-methylhexahydro-1H-spiro[naphthalene-2,2' -[1,3]dioxolane]-5(3H)-one.

Intermediate 11

5-(Chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene

first step

Methyl 6-carbonyloctahydro-1H-indene-5-carboxylate

The compound dimethyl carbonate (0.45g, 5.00mol) and tetrahydrofuran (10mL) were mixed, under argon protection, sodium hydride (0.08g, 2.00mol, 60% mineral oil dispersion) was added in batches, and stirred at 50 ° C for 5 minutes . Then, octahydro-5H-inden-5-one (synthesis reference: Advanced Synthesis & Catalysis, 360(20), 3924-3929; 2018) (0.14 g, 1.00 mol) was added, and the mixture was stirred at 70° C. for 2 hours. The mixture was 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 x 2). The combined organic phases were washed with saturated brine (50 mL×2). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was de-solubilized 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 obtain the target product, methyl 6-carbonyloctahydro-1H-indene-5-carboxylate (0.07g, yellow liquid). Yield: 37%.

¹ H NMR (400MHz, CDCl ₃ ) δ3.76(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).

second step

Methyl 6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carboxylate

Compound 6-carbonyloctahydro-1H-indene-5-carboxylic acid methyl ester (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,N-dimethylformamide (3 mL) and tetrahydrofuran (3 mL) were mixed and stirred at 45 °C for 15 under argon Hour. Cooled to room temperature, the mixture was 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 x 2). The combined organic phases were washed with saturated brine (50 mL×2). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was de-solubilized 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 obtain the target product 6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7, 7a-Hexahydro-1H-indene-5-carboxylic acid methyl ester (37 mg, yellow liquid). Yield: 57%. MS m/z(ESI): 329[M+1];

third step

6-(4-Chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carboxylic acid methyl ester

Compound (6-(((trifluoromethyl)sulfonyl)oxo)-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carboxylic acid methyl ester (37 mg, 0.20 mmol) , 4-chlorophenylboronic acid (50 mg, 0.30 mmol), ethylene glycol dimethyl ether (4 mL), methanol (2 mL) were mixed, and tetrakis(triphenylphosphine) palladium (20 mg, 0.02 mmol) was added under argon protection, Cesium fluoride (80 mg, 0.50 mmol) was reacted at 70° C. for 15 hours under argon protection. Cooled to room temperature, the mixture was quenched with water (25 mL) and ethyl acetate (25 mL). Separate the organic phase and use ethyl acetate for the aqueous phase. Ester (25mL×2) extraction. Combined organic phases were washed with saturated brine (50mL×2). Dry with anhydrous sodium sulfate, filter to remove desiccant, filtrate was removed under reduced pressure to obtain crude product, which was passed through silica gel column (petroleum ether/ Ethyl acetate=30:1) Purification to obtain the target product 6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carboxylic acid methyl ester (40mg, yellow liquid). Yield: 65%.

MS m/z(ESI): 291&293[M+1];

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.26 (d, J=8.0 Hz, 2H), 7.06 (d, J=8.0 Hz, 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).

the fourth step

(6-(4-Chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methanol

Compound 6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene-5-carboxylic acid methyl ester (40 mg, 0.17 mmol) and tetrahydrofuran (3 mL) were mixed, The temperature was lowered to -15°C in an ice-salt bath, and lithium aluminum hydride (10 mg, 0.30 mmol) was slowly added in batches. After the addition, the mixture was naturally raised to room temperature and stirred for 3 hours. Quench by addition of hydrochloric acid (1 M, 0.30 mmol, 0.3 mL). It was extracted with dichloromethane (25 mL×3), and the combined organic phases were washed with saturated brine (25 mL×3). It was dried with anhydrous sodium sulfate, and the drying agent was removed by filtration. The filtrate was desolvated under reduced pressure to obtain the target product (6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methanol (40mg, pale yellow liquid) , Yield: 98%. MS m/z(ESI): 245&247[M-17].

the fifth step

5-(Chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene

Compound (6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methanol (40 mg, 0.13 mmol), thionyl chloride (0.1 mL) and dichloromethane (3 mL), and stirred at room temperature for 1 hour. Precipitate under reduced pressure to obtain the target product 5-(chloromethyl)-6-(4-chlorophenyl)-2,3,3a,4,7,7a-hexahydro-1H-indene (40mg, pale yellow liquid) ,Crude. MS m/z(ESI): 281, 283 & 285 [M+1].

Intermediate 12

(R)-Methyl 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(3-methylpiperazin-1-yl)benzoate

(R)-Methyl 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(3-methylpiperazin-1-yl)benzoate

Compound 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-fluorobenzoic acid methyl ester (synthetic 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) was mixed. Stir at 60°C for 16 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (60 mL x 2). The combined organic phases were washed with saturated brine (50 mL×3). The organic phase was dried with anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolvation under reduced pressure. Purification by column chromatography (dichloromethane/methanol=90:10) gave the target product (R)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-( Methyl 3-methylpiperazin-1-yl)benzoate (2.55 g, yellow solid). Yield: 70%.

MS m/z(ESI): 367[M+1];

¹ H NMR (400 MHz, CDCl ₃ ) 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=1.6Hz,1H),6.34(d,J=2.4Hz,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).

Example 1

2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a- Methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(( (Tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide

first step

2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a- Methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoic acid methyl ester

The compound (3aR,7aS)-5-(chloromethyl)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-indene ( Intermediate 4) (1.01 g, 3.44 mmol), methyl 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(piperazin-1-yl)benzoate Ester (synthetic reference: WO2010138588A2) (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. It was cooled to room temperature, diluted with water (100 mL), extracted with ethyl acetate (100 mL×2), and the combined organic phases were washed with saturated brine (100 mL). The organic phase was dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the filtrate was removed under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=1:1) to obtain the target product 2-((1H -pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2, Methyl 3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoate (0.95 g, white solid). Yield: 45%.

MS m/z(ESI):611&613[M+1];

¹ H NMR (400 MHz, CDCl ₃ ) 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).

second step

(2-((1H-Pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a -Methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoic acid

2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a -methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoate (0.95 g, 1.56 mmol), hydrogen Sodium oxide (0.30 g, 7.50 mmol), ethanol (20 mL) and water (1 mL) were mixed and stirred at 80°C for 1 hour. The solution was removed under reduced pressure, and the residue was added with water (30 mL) and hydrochloric acid (1 M, 15 mL). It was extracted with dichloromethane (30 mL×3), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the drying agent was removed by filtration. Precipitate under reduced pressure to obtain the target product (2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4 -chlorophenyl)-3a-methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoic acid (0.90 g, white solid), yield: 97%. MS m/z (ESI): 597&599[M+1];

third step

2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a- Methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(( (Tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide

(2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)- 3a-Methyl-2,3,3a,4,7,7a-hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)benzoic acid (0.50 g, 1.00 mmol), 3- Nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzenesulfonamide (0.44 g, 1.40 mmol) (Synthesis reference: WO 2018041248 A1), 1-ethyl-( 3-Dimethylaminopropyl)carbodiimide hydrochloride (0.38g, 2.00mmol), 4-dimethylaminopyridine (0.12g, 1.00mmol), triethylamine (0.20g, 2.00mmol) It was mixed with dichloromethane (60 mL) and stirred at room temperature for 16 hours. Water (50 mL) was added to dilute, the organic phase was separated, the aqueous phase was extracted with dichloromethane (50 mL×2), and the combined organic phases were washed with saturated brine (50 mL). Dry with anhydrous sodium sulfate, filter, and remove the filtrate under reduced pressure to obtain the crude product. The residue is purified by flash column chromatography (dichloromethane/methanol=97:3) to obtain the target product 2-((1H-pyrrolo[2] ,3-b]pyridin-5-yl)oxo)-4-(4-(((3aR,7aS)-6-(4-chlorophenyl)-3a-methyl-2,3,3a,4 ,7,7a-Hexahydro-1H-inden-5-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4 -yl)methyl)amino)phenyl)sulfonyl)benzamide 1 (0.27 g, pale yellow solid), yield: 30%.

MS m/z(ESI):894&896[M+1];

¹ H NMR (400MHz, CDCl ₃ ) 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).

The synthetic procedures of Examples 2 to 31 refer to the procedure of Example 1.

biological experiment

Testing for Inhibition of BCL-2 Biological Activity

Evaluation of the Effects of Compounds of the Invention on BCL-2 Biological Activity Using Fluorescence Polarization Experiments

The experimental method is outlined as follows:

The effect of compounds on the biological activity of BCL-2 was evaluated by detecting the effect of the compounds on the binding activity of BCL-2 and leukemia pro-apoptotic protein (BIM) using an affinity assay method based on the principle of fluorescence polarization. The reaction buffer contains the following components: PBS (pH 7.4, 3 mM Na ₂ HPO ₄ , 155 mM NaCl, 1 mM KH ₂ PO ₄ ), 1 mM DTT; human recombinant Bcl-2 protein (Cat. No. 10195-H08E) was purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd., diluted to 5nM with reaction buffer; FITC-labeled BIM polypeptide was purchased from Nanjing GenScript Biotechnology Co., Ltd. and diluted to 5nM with reaction buffer.

Compounds were dissolved and diluted to 0.1, 1, 10 μM in 100% DMSO, followed by 4-fold serial dilutions with DMSO to a minimum concentration of 0.0061, 0.061, 0.61 nM, each concentration point being diluted 50-fold with reaction buffer.

Add 3 μL of compound solution and 12 μL of BCL-2 solution to the black 384-well assay plate, mix well, and incubate at room temperature for 15 minutes. Subsequently, 15 μL of FITC-BIM solution was added, and the reaction mixture was incubated at room temperature for 30 minutes in the dark, and then the fluorescence polarization was detected on an Envision multi-plate reader (Perkin Elmer) immediately, with 480 nm as the excitation wavelength and 535 nm as the emission wavelength. In this experiment, the group without BCL-2 protein was used as a negative control (100% inhibition), and the group with BCL-2 protein but no compound was added as a positive control (0% inhibition). The percent inhibition of BCL-2 affinity by a compound can be calculated using the following formula:

Compound _IC50 values were calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software by the following formula:

Y=Bottom+(Top-Bottom)/(1+10^((logIC ₅₀ -X)*slope factor))

where Y is the percentage of inhibition, X is the logarithm of the concentration of the compound to be tested, Bottom is the maximum percentage of inhibition, Top is the minimum percentage of inhibition, and slope factor is the slope coefficient of the curve.

Testing for Inhibition of BCL-XL Biological Activity

Evaluation of the Effects of Compounds of the Invention on BCL-XL Biological Activity Using Fluorescence Polarization Experiments

The experimental method is outlined as follows:

The effect of compounds on the biological activity of BCL-XL was evaluated by detecting the effect of the compounds on the binding activity of BCL-XL and BIM using an affinity assay method based on the principle of fluorescence polarization. The reaction buffer contains the following components: PBS (pH 7.4, 3 mM Na ₂ HPO ₄ , 155 mM NaCl, 1 mM KH ₂ PO ₄ ), 1 mM DTT; human recombinant Bcl-XL protein (Cat. No. 10455-H08E) was purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd., diluted to 10 nM with reaction buffer; FITC-labeled BIM polypeptide was purchased from Nanjing GenScript Biotechnology Co., Ltd. and diluted to 10 nM with reaction buffer.

Compounds were dissolved and diluted to 1 [mu]M in 100% DMSO, followed by 4-fold serial dilutions in DMSO to a minimum concentration of 0.061 nM, each concentration point being diluted 50-fold with reaction buffer.

Add 3 μL of compound solution and 12 μL of BCL-XL solution to the black 384-well assay plate, mix well, and incubate at room temperature for 15 minutes. Subsequently, 15 μL of FITC-BIM solution was added, and the reaction mixture was incubated at room temperature for 30 minutes in the dark, and then the fluorescence polarization was detected on an Envision multi-plate reader (Perkin Elmer) immediately, with 480 nm as the excitation wavelength and 535 nm as the emission wavelength. In this experiment, the group without BCL-XL protein was used as a negative control (100% inhibition), and the group with BCL-XL protein but no compound was added as a positive control (0% inhibition). The percent inhibition of BCL-XL affinity by a compound can be calculated using the following formula:

Compound _IC50 values were calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software by the following formula:

Y=Bottom+(Top-Bottom)/(1+10^((logIC ₅₀ -X)*slope factor))

where Y is the percentage of inhibition, X is the logarithm of the concentration of the compound to be tested, Bottom is the maximum percentage of inhibition, Top is the minimum percentage of inhibition, and slope factor is the slope coefficient of the curve.

Determination of the half effective inhibitory concentration _IC50 of RS4;11 cells (acute lymphoblastic leukemia cells)

The effect of the compounds of the present invention on the proliferation of RS4;11 cells was evaluated using a luminescence cell viability assay.

The experimental method is outlined as follows:

Using the CellTilter-Glo (CTG) Assay Kit, by using a unique, highly sensitive and stable luciferase to detect ATP, a key indicator of the metabolism of living cells, the luminescent signal generated in the assay and the viable cells in the medium The number is proportional to the number, so as to detect the cell proliferation status of RS4;11.

CellTilter-Glo reagent (Promega, G7572) is composed of CellTilter-Glo lyophilized powder and CellTilter-Glo buffer. When using, the lyophilized powder can be dissolved in the buffer.

RS4;11 cells (ATCC, CRL-1873) were cultured in RPMI1640 complete medium (Thermofisher, 72400-047) containing 10% FBS (GBICO, 10099-141) and 100 units/ml penicillin-streptomycin (Thermofisher, 15140122) ), when the cells covered 80-90% in the culture vessel, they were digested with 0.25% trypsin (containing EDTA) (Thermofisher, Cat. No. 25200056), and then planted in a white 384-well plate (Thermofisher, Cat. No. 164610), and then The 384-well plate was incubated overnight in a 37°C, 5% _CO2 incubator. Compounds were dissolved and diluted to 5 mM in 100% DMSO, followed by 4-fold serial dilutions with DMSO to a minimum concentration of 0.061 μM, and each concentration point was further diluted 50-fold with FBS-free RPMI1640 medium. If the compound has a very low _IC50 value, the starting concentration of the compound can be reduced. After overnight, 3 μL of the compound diluted in medium was added to each well, and the mixture was gently centrifuged to mix. The group without cells was used as a negative control (100% inhibition), and the group added with 0.2% DMSO was used as a positive control (0% inhibition). The 384-well plate was placed in an incubator at 37°C and 5% CO ₂ for continued cultivation. After 48 hours, it was taken out and equilibrated to room temperature. 15 μL of CTG reagent was added to each well, and it was gently shaken on a shaker for 3 minutes to ensure sufficient cell lysis. The luminescent signal was allowed to stabilize for 10 minutes and then read with EnVision (Perkin Elmer).

The percent inhibition of RS4;11 cell proliferation by a compound can be calculated using the following formula:

Percent inhibition = 100-100*(signal compound-signal negative control)/(signal positive control-signal negative control)

Compound _IC50 values were calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software by the following formula:

Y=Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*slope factor)) where Y is the inhibition percentage, Bottom is the bottom plateau of the curve (the bottom plateau value of the S-shaped curve), Top is the top plateau of the curve (the top plateau value of the S-shaped curve), and X is the logarithm of the concentration of the tested compound.

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

Table 1: Detection results of BCL-2 and BCL-XL protein activity

Compound number FP BCL-2 IC ₅₀ (nM) 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 twenty one 19.53 >1000 twenty two 7.14 >1000 twenty three 2.11 >1000 twenty four 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 .

Table 2: RS4;11 cell viability assay results

Compound number 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

twenty three 14 twenty four 16 25 37 26 4.5 27 16 28 5.8 29 6 30 26 31 13 Venetoclax 5.5 .

From the above experimental results, it can be known that the compounds of the examples of the present invention can effectively and selectively inhibit the activity of BCL-2, and the inhibition of BCL-XL is weak. It 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, brain glial tumor etc. And avoid toxic side effects caused by BCL-XL inhibition, such as thrombocytopenia. Some compounds can also effectively inhibit RS4;11 acute lymphocyte proliferation. It has a strong inhibitory effect on malignant blood diseases such as acute lymphoblastic leukemia.

It will be apparent to those skilled in the art that the present disclosure is not limited to the above-described illustrative embodiments, and that it may be embodied in other specific forms without departing from the essential characteristics of the present disclosure. Therefore, it is intended that these embodiments be considered in all respects to be illustrative and non-restrictive, reference should be made to the appended claims, rather than the above-described embodiments, and therefore within the meaning and scope of equivalency of the claims All changes are included.

Claims (10)

A compound of formula (I), its isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt,

in: X ¹ is selected from optionally substituted C3-C6 cycloalkyl or optionally substituted 3-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein said any The selected substituent is selected from hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, 3-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S; preferably , X ¹ is selected from optionally substituted C5-C6 cycloalkyl or optionally substituted 5-6-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein the Optional substituents are selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, 4- or 5-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S; further preferably , X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, and morpholinyl, wherein the optional Substituents are selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl; most preferably, X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4- base, (S)-1,4-dioxan-2-yl, (R)-1,4-dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxa Cyclobutan-3-yl)piperidin-4-yl, (S)-4-(oxetan-3-yl)morpholin-2-yl; X ² is selected from 5-6 membered heterocycloalkylene containing 1 or 2 heteroatoms selected from N, O, S, wherein the heterocycloalkylene is optionally surrounded by 1 or 2 C1-C4 alkyl groups or halogen substitution; preferably, X ² is selected from a 6-membered heterocycloalkylene group containing 2 N atoms, wherein the heterocycloalkylene group is optionally substituted with 1 or 2 C1-C4 alkyl groups; further preferably , X ² is selected from

wherein the piperazinide group is optionally substituted by 1 C1-C4 alkyl group; further preferably, X ² is selected from

wherein the piperazinylene group is optionally substituted with ¹ methyl; most preferably, X is selected from

R ⁰ is selected from hydrogen, halogen; preferably, R ⁰ is selected from hydrogen, fluorine, chlorine; most preferably, R ⁰ is selected from hydrogen, fluorine; R ¹ and R ² are each independently selected from hydrogen and C1-C6 alkyl; preferably, R ¹ and R ² are each independently selected from hydrogen and C1-C4 alkyl; more preferably, R ¹ and R ² are each independently is selected from hydrogen, methyl, ethyl; most preferably, R ¹ , R ² are each independently selected from hydrogen, methyl; n is selected from 1-4; preferably, n is selected from 1, 3 or 4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein: X ¹ is selected from optionally substituted C5-C6 cycloalkyl or optionally substituted 5-6 membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S, wherein any of said The selected substituent is selected from hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, 4- or 5-membered saturated heterocyclic group containing 1 or 2 heteroatoms selected from N, O, S; X ² is selected from a 6-membered heterocycloalkylene group containing 2 N atoms, wherein the heterocycloalkylene group is optionally substituted with 1 or 2 C1-C4 alkyl groups; R ⁰ is selected from hydrogen, halogen; R ¹ and R ² are each independently selected from hydrogen, C1-C6 alkyl; n is selected from 1-4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein: X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, morpholinyl, wherein the optionally substituted group is selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl; X ² is selected from

wherein the piperazinide group is optionally substituted with 1 C1-C4 alkyl group; R ⁰ is selected from hydrogen, halogen; R ¹ and R ² are each independently selected from hydrogen, C1-C4 alkyl; n is selected from 1-4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein: X ¹ is selected from optionally substituted cyclohexyl or optionally substituted tetrahydropyranyl, 1,4-dioxanyl, piperidinyl, morpholinyl, wherein the optionally substituted group is selected from hydroxyl, C1-C4 alkyl, oxetanyl or tetrahydrofuranyl; X ² is selected from

wherein the piperazinide group is optionally substituted with 1 C1-C4 alkyl group; R ⁰ is selected from hydrogen, fluorine, chlorine; R ¹ and R ² are each independently selected from hydrogen, C1-C4 alkyl; n is selected from 1, 3 or 4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein: X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4-yl, (S)-1,4-dioxan-2-yl, (R)-1,4 -Dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxetan-3-yl)piperidin-4-yl, (S)-4-(oxetane alk-3-yl)morpholin-2-yl; X ² is selected from

wherein the piperazinylene group is optionally substituted with 1 methyl; R ⁰ is selected from hydrogen, fluorine; R ¹ and R ² are each independently selected from hydrogen, methyl, and ethyl; n is selected from 1, 3 or 4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt, wherein: X ¹ is selected from (1r,4r)-1-hydroxy-1-methylcyclohexane-4-yl, (S)-1,4-dioxan-2-yl, (R)-1,4 -Dioxan-2-yl, tetrahydropyran-4-yl, 1-(oxetan-3-yl)piperidin-4-yl, (S)-4-(oxetane alk-3-yl)morpholin-2-yl; X ² is selected from

R ⁰ is selected from hydrogen, fluorine; R ¹ and R ² are each independently selected from hydrogen and methyl; n is selected from 1, 3 or 4. A compound of formula (I) according to claim 1, its isomer, prodrug, solvate, stable isotope derivative or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

Use of a compound according to any one of claims 1 to 7, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use as a BCL-2 inhibitor . A compound according to any one of claims 1 to 7, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, in the manufacture of a compound for the treatment or prophylaxis of BCL-2 mediated Use in the medicament of a related disease such as a tumor selected from the group consisting of hematological malignancies including acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, brain glial tumor. A pharmaceutical composition containing a compound according to any one of claims 1-7 or an isomer, prodrug, solvate, stable isotopic derivative or pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents, and excipients.