Sulfonylbenzamide derivatives and their complexes, their preparation method and applications

Abstract

The present invention provides sulfonylbenzamide derivatives and conjugates thereof, their preparation methods, and their applications. Specifically, the present invention provides sulfonylbenzamide derivatives and conjugates thereof having the structure represented by formula (D), their preparation methods, pharmaceutical compositions containing them, and their use in preparing drugs for treating cancer by receptor modulation. The substituents in formula (D) are as defined in the specification. TIFF2023533735000122.tif8862

Description

[Technical Field]

[0001] The present disclosure relates to sulfonylbenzamide derivatives and their ligand-drug conjugates (ADCs). Specifically, the present disclosure relates to sulfonylbenzamide derivatives having novel structures and antibody-drug conjugates thereof, methods for preparing the same, pharmaceutical compositions containing the conjugates, and uses of the conjugates or pharmaceutical compositions. [Background technology]

[0002] One of the key features that distinguishes tumor cells from normal cells is that inhibition of cell apoptosis provides a greater survival advantage, and BCL-2 family proteins are key regulators of the inhibition of apoptosis.

[0003] BCL-2 family proteins are primarily found in the mitochondrial membrane and can be divided into two types based on their function: anti-apoptotic proteins and pro-apoptotic proteins. Anti-apoptotic proteins include BCL-2, BCL-xL, BCL-w, and MCL-1. Pro-apoptotic proteins include Bax, Bak, and BH3-only proteins. When activated, Bax and Bak form a multimer cavity, which increases the permeability of the mitochondrial membrane and promotes the release of cytochrome C and other proteins into the cytoplasm, leading to cell death. BH3-only proteins contain only a BH3 domain. In living cells, BH3-only proteins (e.g., Bim) bind to anti-apoptotic proteins. When cells are subjected to external pressure, the binding equilibrium is broken, and BH3-only proteins are released and bind to BAX in the mitochondria, promoting BAX / BAK multimerization and promoting the release of cytochrome C and SMAC into the cytoplasm, activating downstream apoptotic pathways.

[0004] Previous preclinical data showed that the BCL2 / BCL-xL dual-targeting inhibitor ABT263 is not only effective against hematological malignancies, but also has excellent inhibitory effects on solid tumors, particularly small cell lung cancer. However, ABT263 showed platelet toxicity in clinical trials, limiting its efficacy and resulting in the termination of clinical trials.

[0005] To address clinical needs, there is still a need to explore dual-targeted BCL2 / BCL-xL inhibitors that can be used as monotherapy, in combination, or in ADCs. Summary of the Invention

[0006] The present disclosure provides a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof: [ka] Among them, R 1 is selected from a deuterium atom, a hydrogen atom, an alkyl group, and a deuterated alkyl group; R 2 is selected from alkyl groups and hydroxyalkyl groups, Or R 1 and R 2 together with the atom to which they are linked form a heterocyclyl group, said heterocyclyl group being optionally further substituted with substituents selected from halogen, hydroxy, alkyl, hydroxyalkyl, alkoxy and cycloalkyl groups; R 3 is selected from a hydrogen atom, a hydroxyalkyl group, an alkyl group, a deuterated alkyl group, a cycloalkyl group, and a cycloalkylalkyl group; R 4 is selected from a hydrogen atom, a halogen, a deuterated alkyl group, and an alkyl group; R 5 is selected from halogen and haloalkyl groups; R 6 is selected from an alkyl group, an amino group, a hydroxy group, and an alkoxy group; R7 is selected from a hydrogen atom, a halogen, an alkyl group, a deuterated alkyl group, a hydroxy group, and an alkoxy group; R 8 , R 9 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 8 and R 9 together with the atom to which they are connected form a cycloalkyl group, R 10 , R 11 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 10 and R 11 together with the atom to which they are connected form a cycloalkyl group, R 12 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 13 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 14 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; r is selected from 0, 1, 2 and 3; As a condition, R 1 and R 2 do not form a heterocyclyl group together with the atoms to which they are attached; 5 is a haloalkyl group.

[0007] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 1 is a hydrogen atom, R 2 is an alkyl group.

[0008] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 1 and R 2 form a heterocyclyl group together with the atom to which it is linked, preferably the heterocyclyl group optionally further containing 1 to 3 heteroatoms selected from nitrogen atoms or oxygen atoms, more preferably the R 1 and R 2 together with the atom(s) linked thereto form a 3- to 6-membered heterocyclyl group, preferably the 3- to 6-membered heterocyclyl group optionally further containing 1 to 3 heteroatoms selected from nitrogen atoms or oxygen atoms, most preferably R 1 and R 2 together with the atoms to which they are attached form a pyrrolidinyl group.

[0009] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by the general formula (DI) or a pharmaceutically acceptable salt thereof, [ka] wherein m is selected from 0, 1, 2, and 3; r, R 3 ~R 14 is as defined in general formula (D).

[0010] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by general formula (D-II) or a pharmaceutically acceptable salt thereof, [ka] wherein m is selected from 0, 1, 2, and 3; r, R 3 ~R 11 is as defined in general formula (D).

[0011] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by general formula (D-III) or a pharmaceutically acceptable salt thereof: [ka] wherein m is selected from 0, 1, 2, and 3; r, R 3 ~R 11 is as defined in general formula (D).

[0012] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by general formula (D-IV) or a pharmaceutically acceptable salt thereof, [ka] wherein m is selected from 0, 1, 2, and 3; R 3 ~R 9 is as defined in general formula (D).

[0013] In some other embodiments of the present disclosure, the compound is represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein m is 2.

[0014] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 5 is a chlorine atom.

[0015] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 5 is a trifluoromethyl group.

[0016] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R3 is selected from a hydrogen atom, a hydroxyalkyl group, and an alkyl group, preferably a hydroxyalkyl group; C 1-6 Hydroxyalkyl groups are more preferred.

[0017] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by the general formula (DV) or a pharmaceutically acceptable salt thereof, [ka] wherein t is selected from 0, 1, 2 and 3, preferably 1; r, R 1 , R 2 , R 4 , R 6 ~R 14 is as defined in general formula (D).

[0018] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by general formula (D-VI) or a pharmaceutically acceptable salt thereof, [ka] wherein t is selected from 0, 1, 2 and 3, preferably 1; r, R 1 , R 2 , R 4 , R 6 ~R 11 is as defined in general formula (D).

[0019] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof is a compound represented by general formula (D-VII) or a pharmaceutically acceptable salt thereof, [ka] wherein t is selected from 0, 1, 2 and 3, preferably 1; r, R 1 ~R 11 is as defined in general formula (D).

[0020] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 4 is selected from a hydrogen atom and an alkyl group, and 1-6 An alkyl group is preferred, and a hydrogen atom is more preferred.

[0021] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 6 is selected from a hydroxy group and an alkoxy group, and C 1-6 An alkoxy group is preferred, and a hydroxy group is more preferred.

[0022] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 7 , R 8 and R 9 is a hydrogen atom.

[0023] In some other embodiments of the present disclosure, there is provided a compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein the above R 10 and R 11 are each independently selected from a hydrogen atom and an alkyl group, and 1-6 An alkyl group is preferred, and a hydrogen atom is more preferred.

[0024] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) or a pharmaceutically acceptable salt thereof, wherein r is 0 or 1, and 0 is preferred.

[0025] In some other embodiments of the present disclosure, the compound represented by the above general formula (D) includes, but is not limited to, the following compounds or pharmaceutically acceptable salts thereof: [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] .

[0026] The present disclosure further provides a compound represented by general formula (DA1) or (DB1) or a pharmaceutically acceptable salt thereof: [ka] Among them, G1 is an amino protecting group, preferably Boc; m is selected from 0, 1, 2 and 3; r, R 4 ~R 11 is as defined in general formula (D).

[0027] In some other embodiments of the present disclosure, a compound represented by the above general formula (DA1) or (DB1) or a pharmaceutically acceptable salt thereof is [ka] It is any one compound selected from:

[0028] The present disclosure further provides a method for preparing a compound represented by general formula (D-III) or a pharmaceutically acceptable salt thereof, the method comprising: [ka] a compound of general formula (DA1) or a pharmaceutically acceptable salt thereof is subjected to a deprotection reaction under acidic conditions to obtain a compound of general formula (D-III) or a pharmaceutically acceptable salt thereof, wherein preferably, the reagent used under the acidic conditions is HCl; Among them, G1 is an amino protecting group, preferably Boc; m is selected from 0, 1, 2 and 3; R 3 is a hydrogen atom, r, R 4 ~R 11 is as defined in general formula (D).

[0029] The present disclosure further provides a compound represented by general formula (DA2) or (DB2) or a pharmaceutically acceptable salt thereof: [ka] Among them, m is selected from 0, 1, 2 and 3; r, R 4 ~R 11 is as defined in general formula (D).

[0030] In some other embodiments of the present disclosure, a compound represented by the above general formula (DA2) or (DB2) or a pharmaceutically acceptable salt thereof is [ka] It is any one compound selected from:

[0031] The present disclosure further provides a method for preparing a compound represented by general formula (D-III) or a pharmaceutically acceptable salt thereof, the method comprising: [ka] a compound of general formula (DA2) or a pharmaceutically acceptable salt thereof is subjected to a substitution reaction under basic conditions to obtain a compound of general formula (D-III) or a pharmaceutically acceptable salt thereof, wherein the reagent used under the basic conditions is preferably triethylamine; Among them, m is selected from 0, 1, 2 and 3; R 3 is a hydroxyalkyl group, r, R 4 ~R 11 is as defined in general formula (D).

[0032] The present disclosure further provides a compound represented by general formula (DA3), (DB3) or (DC3) or a pharmaceutically acceptable salt thereof: [ka] Among them, G2 is a hydroxy protecting group, preferably TBS; t is selected from 0, 1, 2 and 3; r, R 1 , R 2 , R 4 ~R 11 is as defined in general formula (D).

[0033] In some other embodiments of the present disclosure, a compound represented by the above general formula (DA3), (DB3) or (DC3) or a pharmaceutically acceptable salt thereof is [ka] It is any one compound selected from:

[0034] The present disclosure further provides a method for preparing a compound represented by general formula (D-VI) or a pharmaceutically acceptable salt thereof, the method comprising: [ka] a compound of general formula (DA3) or a pharmaceutically acceptable salt thereof is subjected to a deprotection reaction under basic conditions to obtain a compound of general formula (D-VI) or a pharmaceutically acceptable salt thereof, wherein the reagent for the basic conditions is tetrabutylammonium fluoride; Among them, G2 is a hydroxy protecting group, preferably TBS; t is selected from 0, 1, 2 and 3; r, R 1 , R 2 , R 4 ~R 11 is as defined in general formula (D).

[0035] The present disclosure further provides a ligand-drug conjugate comprising a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof: [ka] Among them, R 1 is selected from a deuterium atom, a hydrogen atom, an alkyl group, and a deuterated alkyl group; R 2 is selected from alkyl groups and hydroxyalkyl groups, Or R 1 and R 2together with the atom to which they are linked form a heterocyclyl group, said heterocyclyl group being optionally further substituted with substituents selected from halogen, hydroxy, alkyl, hydroxyalkyl, alkoxy and cycloalkyl groups; R 3a is a bond or -(CH2) t -CH2-O-, wherein the alkyl terminus is connected to the N atom of the drug moiety and the -O- terminus is connected to the linker; R 4 is selected from a hydrogen atom, a halogen, a deuterated alkyl group, and an alkyl group; R 5 is selected from halogen and haloalkyl groups; R 6 is selected from an alkyl group, an amino group, a hydroxy group, and an alkoxy group; R 7 is selected from a hydrogen atom, a halogen, an alkyl group, a deuterated alkyl group, a hydroxy group, and an alkoxy group; R 8 , R 9 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 8 and R 9 together with the atom to which they are connected form a cycloalkyl group, R 10 , R 11 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 10 and R 11 together with the atom to which they are connected form a cycloalkyl group, R 12 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 13 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 14is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; r is selected from 0, 1, 2 and 3; t is selected from 0, 1, 2 and 3; The wavy line indicates a covalent bond to a linker unit or ligand.

[0036] In some other embodiments of the present disclosure, there is provided a ligand-drug conjugate comprising a structure represented by the above formula (-D) or a pharmaceutically acceptable salt thereof, wherein the above formula (-D) is selected from the structures represented by the formulas (-DI) and (-DII), [ka] wherein t is selected from 0, 1, 2, and 3; Wavy line, r, R 1 , R 2 , R 4 ~R 14 is as defined in the general formula (-D).

[0037] In some other embodiments of the present disclosure, there is provided a ligand-drug conjugate comprising a structure represented by formula (-D) above, or a pharmaceutically acceptable salt thereof, wherein formula (-D) is selected from the structures represented by formulas (-DIII), (-DIV) and (-DV), [ka] Among them, t is selected from 0, 1, 2 and 3; m is selected from 0, 1, 2 and 3; Wavy line, r, R 1 , R 2 , R 4 ~R 11 is as defined in general formula (-D), Preferably, in the above formula (-DV), R 1 and R 2 does not form a heterocyclyl group.

[0038] In some other embodiments of the present disclosure, there is provided a ligand-drug conjugate comprising a structure represented by formula (-D) above, or a pharmaceutically acceptable salt thereof, wherein formula (-D) above is selected from structures represented by formulas (-DVI) and (-DVII), [ka] Among them, t is selected from 0, 1, 2 and 3; m is selected from 0, 1, 2 and 3; Wavy line, r, R 5 ~R 9 is as defined in the general formula (-D).

[0039] In some other embodiments of the present disclosure, a ligand-drug conjugate comprising a structure represented by the above formula (-D) or a pharmaceutically acceptable salt thereof, which is a ligand-drug conjugate represented by the general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, [ka] Among them, R 1 is selected from a deuterium atom, a hydrogen atom, an alkyl group, and a deuterated alkyl group; R 2 is selected from alkyl groups and hydroxyalkyl groups, Or R 1 and R 2 together with the atom to which they are linked form a heterocyclyl group, said heterocyclyl group being optionally further substituted with substituents selected from halogen, hydroxy, alkyl, hydroxyalkyl, alkoxy and cycloalkyl groups; R 3a is a bond and -(CH2) t -CH2-O-, R 4 is selected from a hydrogen atom, a halogen, a deuterated alkyl group, and an alkyl group; R 5 is selected from halogen and haloalkyl groups; R 6is selected from an alkyl group, an amino group, a hydroxy group, and an alkoxy group; R 7 is selected from a hydrogen atom, a halogen, an alkyl group, a deuterated alkyl group, a hydroxy group, and an alkoxy group; R 8 , R 9 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 8 and R 9 together with the atom to which they are connected form a cycloalkyl group, R 10 , R 11 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 10 and R 11 together with the atom to which they are connected form a cycloalkyl group, R 12 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 13 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 14 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; r is selected from 0, 1, 2 and 3; t is selected from 0, 1, 2 and 3; n is an integer or decimal number from 1 to 10, Pc is a ligand and L is a linker unit.

[0040] In some other embodiments of the present disclosure, a ligand-drug conjugate comprising a structure represented by the above formula (-D) or a pharmaceutically acceptable salt thereof, which is a ligand-drug conjugate represented by the general formula (Pc-L-DIII), (Pc-L-DIV) and (Pc-L-DV) or a pharmaceutically acceptable salt thereof, [ka] Among them, t is selected from 0, 1, 2 and 3; m is selected from 0, 1, 2 and 3; r, R 1 , R 2 , R 4 ~R 11 , Pc, L, and n are as defined in the general formula (Pc-LD).

[0041] In some other embodiments of the present disclosure, in the above-mentioned ligand-drug conjugate or a pharmaceutically acceptable salt thereof, n is an integer or decimal number of 1 to 8, preferably an integer or decimal number of 2 to 8.

[0042] In some other embodiments of the present disclosure, the ligand-drug conjugate or a pharmaceutically acceptable salt thereof, wherein the Pc is an antibody.

[0043] In some other embodiments of the present disclosure, the ligand-drug conjugate or a pharmaceutically acceptable salt thereof is any one of an anti-HER2 (ErbB2) antibody, an anti-Trop-2 antibody, an anti-CD79b antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD44 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-A33 antibody, an anti-Cripto antibody, an anti-EphA2 antibody, an anti-G250 antibody, an anti-MUCl antibody, an anti-Lewis antibody, an anti-CD105 ... Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-integrin antibody, anti-PSMA antibody, anti-tenascin-C antibody, anti-SLC44A4 antibody and anti-mesothelin antibody, preferably selected from Trastuzumab, Pertuzumab, Nimotuzumab, Enoblituzumab, Emibetuzumab, Inotuzumab, Pinatuzumab, Brentuximab, Gemtuzumab, Bivatuzumab, Lorvotuzumab, cBR96 and Glematumumab.

[0044] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof, wherein the linker unit -L- is -L a -L b -L c - and L a teeth [ka] W is selected from -C 1-6 Alkyl group -, -C 1-6 alkyl-cycloalkyl- and straight-chain heteroalkyl- of 1 to 6 atoms, wherein the heteroalkyl group contains 1 to 3 heteroatoms selected from N, O and S, among which the -C 1-6 Alkyl-, -C 1-6each alkyl-cycloalkyl- or straight-chain-heteroalkyl- of 1 to 6 atoms is independently optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, amino, alkyl, chloroalkyl, deuterated alkyl, alkoxy, and cycloalkyl; L b is a peptide residue or chemical bond consisting of 2 to 7 amino acids, wherein the amino acids are optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, amino, alkyl, chloroalkyl, deuterated alkyl, alkoxy, and cycloalkyl; L c Ha-NR 17 (CR 18 R 19 )t-, -NH-C(R 18 R 19 )-OC(R 20 R 21 )-C(O)-, -NH-R 22 -(CH2) q -OC(O)-, -C(O)NR 17 , -C(O)NR 17 (CH2) q - and a chemical bond, wherein q is an integer from 1 to 6; R 17 is selected from a hydrogen atom, an alkyl group, a haloalkyl group, a deuterated alkyl group, and a hydroxyalkyl group; R 18 and R 19 are the same or different and are each independently selected from a hydrogen atom, a halogen, an alkyl group, a haloalkyl group, a deuterated alkyl group, and a hydroxyalkyl group; R 20 is selected from alkyl groups, cycloalkylalkyl groups, and cycloalkyl groups; R 21 is selected from a hydrogen atom, an alkyl group, and a haloalkyl group; Or R 20 and R 21 together with the carbon atoms connected to them, 3-6 forming a cycloalkyl group, R22 is selected from an aryl group or a heteroaryl group.

[0045] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof is as described above, wherein W is selected from -(CH2)2- and -(CH2)5-.

[0046] In some other embodiments of the present disclosure, the ligand-drug conjugate or a pharmaceutically acceptable salt thereof, wherein L b The peptide residue is an amino acid residue formed by one or more amino acids selected from phenylalanine, glycine, valine, lysine, citrulline, serine, glutamic acid, and aspartic acid, and is preferably a tetrapeptide residue, a dipeptide residue, or a chemical bond, and more preferably a tetrapeptide residue of glycine-glycine-phenylalanine-glycine or a dipeptide residue of valine-citrulline.

[0047] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof, wherein the linker unit L c is -NH-C(R 18 R 19 )-OC(R 20 R 21 )-C(O)-, -NH-R 22 -(CH2) q -OC(O)- or a chemical bond, q is an integer from 1 to 6, and R 22 is selected from aryl groups and heteroaryl groups; Preferably, L c has the following structural formula: [ka] Selected from R 18 and R 19 are the same or different and are each independently selected from a hydrogen atom, a halogen, an alkyl group, a haloalkyl group, a deuterated alkyl group, and a hydroxyalkyl group; R20 is selected from alkyl groups, cycloalkylalkyl groups, and cycloalkyl groups; R 21 is selected from a hydrogen atom, an alkyl group, and a haloalkyl group; Or R 20 and R 21 together with the carbon atoms connected to them, 3-6 Forms a cycloalkyl group.

[0048] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof, wherein the linker unit -L- is -L a - and -L a - is as defined above.

[0049] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof, wherein the linker unit -L- is -L a -L b -L c - and L a teeth [ka] wherein W is selected from -(CH2)2- and -(CH2)5-; L b is selected from a tetrapeptide residue and a dipeptide residue, and is preferably a tetrapeptide residue of glycine-glycine-phenylalanine-glycine or a dipeptide residue of valine-citrulline; L c has the following structural formula: [ka] Selected from R 18 or R 19 are the same or different and are each independently selected from a hydrogen atom, a halogen, an alkyl group, a haloalkyl group, a deuterated alkyl group, and a hydroxyalkyl group; R20 is selected from an alkyl group, a cycloalkylalkyl group, or a cycloalkyl group; R 21 is selected from a hydrogen atom, an alkyl group, or a haloalkyl group; Or R 20 and R 21 together with the carbon atoms connected to them, 3-6 Forms a cycloalkyl group.

[0050] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or pharmaceutically acceptable salt thereof comprises a linking unit -L-, wherein -L- is [ka] Selected from The a-terminus is connected to the ligand Pc, and the b-terminus is connected to the drug terminal R 3a is connected to.

[0051] In some other embodiments of the present disclosure, the Ligand-Drug conjugate or a pharmaceutically acceptable salt thereof, wherein the linker unit -L- is -L a -L b -L c - and L a teeth [ka] wherein W is selected from -(CH2)2- and -(CH2)5-; L b is selected from a tetrapeptide residue and a dipeptide residue, and is preferably a tetrapeptide residue of glycine-glycine-phenylalanine-glycine or a dipeptide residue of valine-citrulline; L c has the following structural formula: [ka] is.

[0052] The present disclosure further provides a compound represented by the general formula (Lu-D) or a pharmaceutically acceptable salt thereof: [ka] Among them, L a’ teeth, [ka] wherein W is selected from -(CH2)2- and -(CH2)5-; L b is a peptide residue or chemical bond consisting of 2 to 7 amino acids, wherein the amino acids are optionally substituted with one or more substituents selected from halogen, hydroxyl, cyano, amino, alkyl, chloroalkyl, deuterated alkyl, alkoxy, and cycloalkyl; L c has the following structural formula: [ka] and R 1 is selected from a deuterium atom, a hydrogen atom, an alkyl group, and a deuterated alkyl group; R 2 is selected from alkyl groups and hydroxyalkyl groups, Or R 1 and R 2 together with the atom to which they are linked form a heterocyclyl group, said heterocyclyl group being optionally further substituted with substituents selected from halogen, hydroxy, alkyl, hydroxyalkyl, alkoxy and cycloalkyl groups; R 3a is a bond and -(CH2) t -CH2-O-, R 4 is selected from a hydrogen atom, a halogen, a deuterated alkyl group, and an alkyl group; R 5 is selected from halogen and haloalkyl groups; R 6is selected from an alkyl group, an amino group, a hydroxy group, and an alkoxy group; R 7 is selected from a hydrogen atom, a halogen, an alkyl group, a deuterated alkyl group, a hydroxy group, and an alkoxy group; R 8 , R 9 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 8 and R 9 together with the atom to which they are connected form a cycloalkyl group, R 10 , R 11 are each independently selected from a hydrogen atom, an alkyl group, a deuterated alkyl group, and a cycloalkyl group, or R 10 and R 11 together with the atom to which they are connected form a cycloalkyl group, R 12 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 13 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; R 14 is selected from a hydrogen atom, a deuterium atom, a deuterated alkyl group, a halogen, a hydroxy group, an alkyl group, a hydroxyalkyl group, an alkoxy group, and a cycloalkyl group; r is selected from 0, 1, 2 and 3; t is selected from 0, 1, 2 and 3.

[0053] In some other embodiments of the present disclosure, the compound represented by the above general formula (Lu-D) or a pharmaceutically acceptable salt thereof is as follows: [ka]

[0054] The present disclosure further provides a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate having a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate represented by general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable vector, diluent, or excipient.

[0055] The present disclosure further provides a use of a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate comprising a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate represented by general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition, in the preparation of a Bcl-2 and / or Bcl-xL inhibitor.

[0056] The present disclosure further provides a use of a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate comprising a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate represented by general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition, in the preparation of a medicament for treating or preventing a tumor.

[0057] The present disclosure further provides a use of a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate comprising a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate represented by general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described above, in the preparation of a medicament for treating and / or preventing cancer, among which the cancers include melanoma, liver cancer, renal cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, nasopharyngeal cancer, Colorectal cancer, colon cancer, rectal cancer, pancreatic cancer, cervical cancer, ovarian cancer, breast cancer, bladder cancer, prostate cancer, leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, acute granulocytic leukemia, mantle cell lymphoma, diffuse large B-cell lymphoma, follicle center lymphoma, non-Hodgkin's lymphoma, T-cell lymphoma, B-cell lymphoma, head and neck squamous cell carcinoma, cervical cancer, thyroid cancer, lymphoma, sarcoma, neuroblastoma, brain tumor, myeloma, astrocytoma and glioma are preferred.

[0058] Another aspect of the present disclosure further relates to a method for treating or preventing cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate comprising a structure represented by formula (-D) or a pharmaceutically acceptable salt thereof, or a ligand-drug conjugate represented by general formula (Pc-LD) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, wherein the cancer is selected from the group consisting of melanoma, liver cancer, renal cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, and melanoma. Preferred are lung cancer, nasopharyngeal cancer, colorectal cancer, colon cancer, rectal cancer, pancreatic cancer, cervical cancer, ovarian cancer, breast cancer, bladder cancer, prostate cancer, leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, acute granulocytic leukemia, mantle cell lymphoma, diffuse large B-cell lymphoma, follicle center lymphoma, non-Hodgkin's lymphoma, T-cell lymphoma, B-cell lymphoma, squamous cell carcinoma of the head and neck, cervical cancer, thyroid cancer, lymphoma, sarcoma, neuroblastoma, brain tumor, myeloma, astrocytoma, and glioma.

[0059] The active compound (e.g., a ligand-drug conjugate described herein, or a pharmaceutically acceptable salt thereof) can be prepared in a form suitable for administration by any suitable route, preferably in a unit dose form or in a form that allows a subject to self-administer a single dose. The unit dose form of the active compound or composition described herein may be a tablet, capsule, cachet, bottled drug solution, drug powder, granules, topical tablet, suppository, reconstituted powder, or liquid formulation.

[0060] The dosage of the active compound or composition used in the therapeutic methods of the present disclosure will generally vary depending on the severity of the disease, the weight of the subject, and the relative potency of the active compound. As a general guide, a suitable unit dose may be from 0.1 mg to 1000 mg.

[0061] The pharmaceutical composition according to the present disclosure may contain one or more additives in addition to the active compound, and the additives may be selected from components such as fillers, diluents, binders, wetting agents, disintegrants, or excipients. The composition may contain 0.1 to 99% by weight of the active compound, depending on the administration route.

[0062] The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as a tablet, a sugar-coated tablet, an external tablet, an aqueous or oily suspension, a dispersible powder or granules, an emulsion, a hard or soft capsule, or a syrup or elixir. Oral compositions can be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain binders, fillers, lubricants, disintegrating agents, or pharmaceutically acceptable wetting agents, and may also contain one or more ingredients selected from sweeteners, flavoring agents, coloring agents, and preservatives to provide a pharmaceutical preparation that is visually and palatably pleasing.

[0063] Aqueous suspensions contain the active substances and excipients suitable for the preparation of aqueous suspensions. Aqueous suspensions may also contain one or more preservatives, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents.

[0064] Oily suspensions may be prepared by suspending the active ingredient in vegetable oil. Oily suspensions may contain thickening agents. In order to provide a palatable preparation, the above-mentioned sweeteners and flavoring agents may be added.

[0065] Pharmaceutical compositions provide the active ingredient in the form of dispersible powders and granules for preparing aqueous suspensions, which may be mixed with one or more dispersing agents, wetting agents, suspending agents, or preservatives by adding water. Other excipients such as sweeteners, flavoring agents, and coloring agents may also be added. These compositions are preserved by adding antioxidants, such as ascorbic acid.

[0066] Pharmaceutical compositions according to the present disclosure may also be in the form of oil-in-water emulsions.

[0067] The pharmaceutical compositions may be in the form of a sterile injectable aqueous solution. Acceptable solvents or vehicles that can be used include water, Ringer's solution, and isotonic sodium chloride solution. The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient may be dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerin and processed to form a microemulsion. The injectable solution or microemulsion can be infused into the subject's bloodstream via local bolus injection. Alternatively, solutions and microemulsions are preferably administered in a manner that maintains a constant, cyclical concentration of the compounds of the present disclosure. To maintain such a constant concentration, a continuous intravenous administration device can be used. An example of such a device is the Deltec CADD-PLUS™ 5400 intravenous pump.

[0068] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension for intramuscular and subcutaneous administration. Such suspensions can be prepared according to known techniques using the above-mentioned suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may be a sterile injectable solution or suspension prepared in a non-toxic parenterally acceptable diluent or solvent. Sterile fixed oils can also be conveniently used as a solvent or suspending medium.

[0069] The compounds of the present disclosure may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid in the rectum, so that it melts in the rectum and releases the drug. Such materials include cocoa butter, glycerol gelatin, hydrogenated vegetable oil, polyethylene glycols of various molecular weights, and mixtures of fatty acid esters of polyethylene glycol.

[0070] As is well known to those skilled in the art, the dosage of a drug depends on many factors, including, but not limited to, the activity of the specific compound used, the age of the subject, the weight of the subject, the health condition of the subject, the behavior of the subject, the diet of the subject, the time of administration, the mode of administration, the excretion rate, the composition of the drug, etc. Furthermore, the optimal treatment method, such as the treatment mode, the daily dose of the general formula compound, and the type of pharmaceutically acceptable salt, can be verified according to conventional treatment plans.

[0071] The present disclosure relates to conjugates for improving the targeting ability of various cancer cells, infectious organisms, and / or treating autoimmune diseases, which conjugates comprise a targeting (binding) moiety and a therapeutic moiety belonging to a drug. The antibody targeting moiety and the compound therapeutic moiety are linked via an intracellularly soluble bond that increases the therapeutic effect.

[0072] For many years, scientists in the field of targeted drug therapy have aimed to utilize monoclonal antibodies (MAbs) to specifically deliver toxic drugs to human cancers. Conjugates of tumor-associated MAbs and suitable toxic drugs have been developed, but these have met with mixed success in treating cancer and have rarely been applied to other diseases, such as infectious diseases and autoimmune disorders. Toxic drugs are the most common chemotherapeutic agents. Currently, there is a need to further develop more effective antibody conjugates with cytosolic linkers for treating cancer, pathogens, and other diseases.

[0073] Detailed Description of the Invention Unless otherwise limited, all technical and scientific terms used herein are consistent with those commonly understood by those skilled in the art. Although any methods and materials similar or equivalent to those described herein can be used to practice or test the present disclosure, the preferred methods and materials are described herein. In describing and claiming the present disclosure, the following terms will be used in accordance with the following definitions.

[0074] When trade names are used in this disclosure, they are intended to include the formulation of the trade name product, the drug and active drug portion of the trade name product.

[0075] The term "ligand" refers to a large molecule compound capable of recognizing and binding to an antigen or receptor associated with a target cell. The function of a ligand is to deliver a drug to a target cell population that is bound by the ligand. These ligands include, but are not limited to, protein hormones, lectins, growth factors, antibodies, or other molecules capable of binding to cells. In an embodiment of the present disclosure, the ligand is represented as Pc, and the ligand can be combined with a linking unit via a heteroatom in the ligand to form a linking bond, and is preferably an antibody or an antigen-binding fragment thereof, and the antibody is selected from a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody, and is preferably a monoclonal antibody.

[0076] The term "drug" refers to a cytotoxic drug or an immunomodulator. Cytotoxic drugs can deliver chemical molecules into tumor cells that are relatively disruptive to their normal growth. In principle, cytotoxic drugs can kill tumor cells if they are present in sufficiently high concentrations. However, due to lack of specificity, they can also cause apoptosis of normal cells while killing tumor cells, resulting in serious side effects. The term also includes, for example, small molecule toxins or enzymatically active toxins derived from bacteria, fungi, plants, or animals, radioactive isotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 and radioactive isotopes of Lu), chemotherapeutic agents, antibiotics, and nucleases. In some embodiments of the present disclosure, the drug is designated D and may be a BCL2 inhibitor, a BCL-xL inhibitor, or a BCL2 / BCL-xL dual-targeted inhibitor.

[0077] The term "linker unit," "linker," or "linking fragment" refers to a chemical structure fragment or bond that is linked at one end to a ligand and at the other end to a drug, and may be linked to another linker and then to the ligand or drug.

[0078] The linker may comprise an extender, a spacer, and an amino acid unit and may be synthesized by methods known in the art, such as those described in US2005-0238649A1. The linker may be a "cleavable linker" that facilitates drug release in cells. For example, an acid-labile linker (e.g., hydrazone), a protease-sensitive (e.g., peptidase-sensitive) linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker (Chari et al., Cancer Research 52:127-131 (1992), U.S. Patent No. 5,208,020) may be used.

[0079] The term "ligand-drug conjugate" refers to a ligand linked to a biologically active drug by a stable linking unit. In the present disclosure, "ligand-drug conjugate" refers to an antibody drug conjugate (ADC), preferably an antibody drug conjugate (ADC), in which a monoclonal antibody or antibody fragment is linked to a biologically active toxic drug by a stable linking unit.

[0080] The three-letter and one-letter codes for amino acids used in this disclosure are as described in J. Biol. Chem, 243, p. 3558 (1968).

[0081] "Antibody" as used in this disclosure is used in the broadest sense herein and encompasses different antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, murine antibodies, chimeric antibodies, humanized antibodies, multispecific antibodies (e.g., bispecific antibodies), and antigen-binding fragments, as long as they exhibit the desired antigen-binding activity and specificity.

[0082] Methods for producing and purifying antibodies and antigen-binding fragments well known in the art are described, for example, in Chapters 5-8 and 15 of Reisenko's Antibody Laboratory Techniques Manual. Antigen-binding fragments can also be prepared by conventional methods. The antibodies or antigen-binding fragments described in this disclosure have one or more human-derived FR regions added to non-human CDR regions by genetic engineering. Human FR germline sequences can be obtained from the ImMunoGeneTics (IMGT) homepage at http: / / imgt.cines.fr by aligning the IMGT human antibody variable region germline gene database with MOE software, or from the Immunoglobulin Journal 2001, ISBN 012441351.

[0083] The term "alkyl group" refers to a saturated aliphatic hydrocarbon group that is a straight-chain or branched-chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms, more preferably an alkyl group containing 1 to 10 carbon atoms, and most preferably an alkyl group containing 1 to 6 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, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl groups, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof.More preferred are lower alkyl groups containing 1 to 6 carbon atoms, and 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, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and 2,3-dimethylbutyl. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available linkage site, and the substituent is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, mercapto groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, and oxo groups.

[0084] The term "heteroalkyl group" refers to an alkyl group containing one or more heteroatoms selected from N, O, or S, wherein alkyl groups are as defined above.

[0085] The term "alkylene group" refers to a saturated, straight-chain or branched-chain aliphatic hydrocarbon group having a residue derived from a parent alkane by removing two hydrogen atoms from the same two carbon atoms or from two different carbon atoms, and is a straight-chain or branched-chain group containing 1 to 20 carbon atoms, preferably 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms, and more preferably 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH-), 1,1-ethylidene (-CH(CH)-), 1,2-ethylidene (-CHCH)-, 1,1-propylidene (-CH(CHCH)-), 1,2-propylidene (-CHCH(CH)-), 1,3-propylidene (-CHCHCHCH-), 1,4-butylidene (-CHCHCHCHCH-), and 1,5-butylidene (-CHCHCHCHCHCH-). The alkylene group may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available linkage site, and preferably the substituents are independently optionally substituted with one or more substituents selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, mercapto groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocyclyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, and oxo groups.

[0086] The term "alkoxy group" refers to -O-(alkyl group) and -O-(unsubstituted cycloalkyl group), where alkyl group and cycloalkyl group are defined above. Non-limiting examples of alkoxy groups include methoxy group, ethoxy group, propoxy group, butoxy group, cyclopropoxy group, cyclobutoxy group, cyclopentyloxy group, and cyclohexyloxy group. An alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably independently one or more groups selected from alkyl group, alkenyl group, alkynyl group, alkoxy group, alkylthio group, alkylamino group, halogen, mercapto group, hydroxy group, nitro group, cyano group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, cycloalkoxy group, heterocycloalkoxy group, cycloalkylthio group, and heterocycloalkylthio group.

[0087] The term "cycloalkyl group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, where the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like, and polycyclic cycloalkyl groups include spirocyclic, fused, and bridged cycloalkyl groups.

[0088] The term "cycloalkylene group" refers to a cycloalkyl group having two residues derived by removing two hydrogen atoms from ring atoms of a parent cycloalkyl group, where the cycloalkyl group is as defined above.

[0089] The term "heterocyclyl group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, of which one or more ring atoms is nitrogen, oxygen, or S(O).m (where m is an integer of 0 to 2), but does not include the -OO-, -OS-, or -SS- ring moieties, with the remaining ring atoms being carbon. Preferably, the cycloalkyl group contains 3 to 12 ring atoms, of which 1 to 4 (e.g., 1, 2, 3, and 4) are heteroatoms. More preferably, the cycloalkyl group contains 3 to 10 ring atoms, more preferably 3 to 8 ring atoms (e.g., 3, 4, 5, 6, 7, and 8), of which 1 to 3 (e.g., 1, 2, and 3) are heteroatoms. Even more preferably, the cycloalkyl group contains 3 to 6 ring atoms, of which 1 to 3 are heteroatoms. Most preferably, the cycloalkyl group contains 5 or 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyl groups include spirocyclic, fused-ring and bridged-ring heterocyclyl groups.

[0090] The term "spiroheterocyclyl group" refers to a 5- to 20-membered polycyclic heterocyclyl group in which the monocyclic rings share one atom (called a spiroatom), in which one or more of the ring atoms is nitrogen, oxygen, or S(O). m (wherein m is an integer of 0 to 2), and the remaining ring atoms are carbon. It may contain one or more double bonds, but none of the rings has a completely conjugated π-electron system. It is preferably 6 to 14-membered, more preferably 7 to 10-membered (e.g., 7, 8, 9, or 10-membered). Depending on the number of spiro atoms shared by the rings, spiroheterocyclyl groups are classified as monospiroheterocyclyl groups, bisspiroheterocyclyl groups, or polyspiroheterocyclyl groups, and preferred are monospiroheterocyclyl groups and bisspiroheterocyclyl groups. More preferred are 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospiroheterocyclyl groups. Non-limiting examples of spiroheterocyclyl groups are: [ka] Includes:

[0091] The term "fused heterocyclyl group" refers to a 5- to 20-membered polycyclic heterocyclyl group in which each ring in the system shares an adjacent pair of atoms with another ring in the system, one or more rings may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system, and one or more ring atoms is nitrogen, oxygen, or S(O) m (wherein m is an integer of 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered (e.g., 7, 8, 9, or 10-membered). Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclyl group, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclyl group. Non-limiting examples of fused heterocyclyl groups are: [ka] Includes:

[0092] The term "bridged heterocyclyl group" refers to a 5- to 14-membered polycyclic heterocyclyl group in which any two rings share two non-directly connected atoms, and may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system, and in which one or more ring atoms is nitrogen, oxygen, or S(O). m (wherein m is an integer of 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered (e.g., 7, 8, 9, or 10-membered). Depending on the number of rings constituting it, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclyl group, preferably a bicyclic, tricyclic, or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclyl groups are: [ka] Includes:

[0093] The heterocyclyl group ring may be fused to an aryl group, heteroaryl group, or cycloalkyl group ring, where the ring connected to the parent structure is a heterocyclyl group, non-limiting examples of which are: [ka] Includes:

[0094] The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably independently one or more groups selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogen atoms, mercapto groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, and oxo groups.

[0095] The term "heterocyclylene group" refers to a heterocyclyl group having two residues derived by removing two hydrogen atoms from the same or different ring atoms of a parent heterocyclyl group, wherein the heterocyclyl group is as defined above.

[0096] The term "aryl group" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π-electron system, preferably 6- to 10-membered, such as phenyl and naphthyl groups, preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, in which the ring connected to the parent structure is an aryl ring, non-limiting examples of which are: [ka] Includes:

[0097] The aryl group may be substituted or unsubstituted. When the aryl group is substituted, the substituents are preferably independently one or more groups selected from an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, an alkylthio group, an alkylamino group, a halogen atom, a mercapto group, a hydroxy group, a nitro group, a cyano group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a cycloalkoxy group, a heterocycloalkoxy group, a cycloalkylthio group, and a heterocycloalkylthio group.

[0098] The term "heteroaryl group" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, where the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5 to 10-membered, more preferably 5 or 6-membered, and examples thereof include furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, and tetrazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, where the ring connected to the parent structure is the heteroaryl ring, and non-limiting examples thereof include: [ka] Includes:

[0099] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably independently one or more groups selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, mercapto groups, hydroxy groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, and heterocycloalkylthio groups.

[0100] The term "amino-protecting group" refers to a group that protects an amino group with an easily removable group so that the amino group is not altered when other sites on the molecule react. Non-limiting examples include 9-fluorenylmethyloxycarbonyl, tert-butoxycarbonyl, acetyl, benzyl, allyl, and p-methoxybenzyl. These groups can be optionally substituted with 1 to 3 substituents selected from halogen, alkoxy, or nitro. The amino-protecting group is preferably 9-fluorenylmethyloxycarbonyl.

[0101] The term "alkenyl group" refers to an alkyl group compound containing a carbon-carbon double bond in the molecule, of which alkyl is as defined above. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from a hydrogen atom, an alkyl group, an alkoxy group, a halogen atom, a haloalkyl group, a haloalkoxy group, a cycloalkyloxy group, a heterocyclyloxy group, a hydroxy group, a hydroxyalkyl group, a cyano group, an amino group, a nitro group, a cycloalkyl group, a heterocyclyl group, an aryl group, and a heteroaryl group.

[0102] The term "alkynyl group" refers to an alkyl group compound containing a carbon-carbon triple bond in the molecule, of which alkyl is defined as above. The alkynyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from a hydrogen atom, an alkyl group, an alkoxy group, a halogen atom, a haloalkyl group, a haloalkoxy group, a cycloalkyloxy group, a heterocyclyloxy group, a hydroxy group, a hydroxyalkyl group, a cyano group, an amino group, a nitro group, a cycloalkyl group, a heterocyclyl group, an aryl group, and a heteroaryl group.

[0103] The term "cycloalkylalkyl group" refers to an alkyl group substituted with one or more cycloalkyl groups, preferably one cycloalkyl group, wherein the alkyl group is as defined above and the cycloalkyl group is as defined above.

[0104] The term "haloalkyl group" refers to an alkyl group substituted with one or more halogens, wherein the alkyl group is as defined above.

[0105] The term "deuterated alkyl group" refers to an alkyl group substituted with one or more deuterium atoms, wherein the alkyl group is as defined above.

[0106] The term "hydroxy" refers to an -OH group.

[0107] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0108] The term "amino group" refers to -NH2.

[0109] The term "nitro group" refers to -NO2.

[0110] The term "amide group" refers to a -C(O)N (alkyl) or (cycloalkyl) group, wherein alkyl and cycloalkyl groups are as defined above.

[0111] The term "carboxylic acid ester group" refers to a -C(O)O(alkyl group) or a -C(O)O(cycloalkyl group), wherein alkyl and cycloalkyl groups are as defined above.

[0112] The abbreviation "Me" in the chemical formula is the methyl group.

[0113] The abbreviation "Ph" in the chemical formula stands for the phenyl group.

[0114] The present disclosure further includes various deuterated forms of the compound of formula (D). Each available hydrogen atom connected to a carbon atom may be independently replaced with a deuterium atom. Those skilled in the art can synthesize the deuterated form of the compound of formula (D) by referring to relevant literature. The deuterated form of the compound of formula (D) may be prepared using commercially available deuterated starting materials or may be synthesized by conventional techniques using deuterated reagents, including, but not limited to, deuterated borane, tritiated borane in tetrahydrofuran, lithium aluminum deuterated hydride, deuterated iodoethane, and deuterated iodomethane.

[0115] "Optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes cases where the event or circumstance occurs and cases where it does not. For example, "a heterocyclyl group optionally substituted with an alkyl group" means that the alkyl group may or may not be present, and the description includes cases where the heterocyclyl group is substituted with an alkyl group and cases where the heterocyclyl group is not substituted with an alkyl group.

[0116] The term "substituted" means that one or more hydrogen atoms in a group, preferably 5 or less, more preferably 1 to 3 hydrogen atoms, are independently replaced with a corresponding number of substituents. Of course, substituents are located only at their chemically feasible positions, and those skilled in the art can determine possible or impossible substitutions (by experiment or theory) without much effort. For example, an amino group or a hydroxy group having free hydrogen may be unstable when bonded to a carbon atom having an unsaturated (e.g., olefinic) bond.

[0117] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein or physiologically / pharmaceutically acceptable salts or prodrugs thereof with other chemical components, and other components such as physiologically / pharmaceutically acceptable vectors and excipients. The pharmaceutical composition is intended to facilitate administration to a living body and contribute to the absorption of the active ingredients to further exert biological activity.

[0118] The term "pharmaceutically acceptable salt" or "medicinal salt" refers to a salt of a ligand-drug conjugate according to the present disclosure or a salt of a compound described herein, which salt is safe and effective when used in a mammalian body and retains the requisite biological activity. The antibody-antibody drug conjugate compounds according to the present disclosure contain at least one amino group and can therefore form salts with acids. Non-limiting examples of medicinal salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogensulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0119] The term "drug loading" refers to the average amount of cytotoxic drugs loaded on each ligand in a conjugate molecule and may be expressed as the ratio of the drug amount to the antibody amount (Drug-to-Antibody Ratio, DAR value). The drug loading ranges from 0 to 12, preferably 1 to 10, cytotoxic drugs (D) linked to each ligand (Pc). In embodiments of the present disclosure, the drug loading is represented by n, which may be, for example, an average value of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, ranging from 0 to 12, preferably 1 to 10, and more preferably 1 to 8, 2 to 8, 2 to 7, 3 to 8, 3 to 7, 3 to 6, 4 to 7, 4 to 6, or 4 to 5. The drug loading of each ADC molecule after the coupling reaction can be characterized and identified by conventional methods, such as UV / visible spectroscopy, mass spectrometry, ELISA, CE-SDS, and HPLC.

[0120] The loading of the ligand-cytotoxic drug conjugate is (1) controlling the molar ratio of the linking reagent to the ligand; (2) controlling the reaction time and temperature; (3) selecting different reaction reagents; The amount of oxygen can be controlled by a number of methods, including but not limited to:

[0121] The preparation of conventional pharmaceutical compositions is set out in the Chinese Pharmacopoeia.

[0122] The term "vector," as used in the present disclosure, refers to a system that can change the drug's entry into and distribution within the human body, control the drug's release rate, and transport the drug to target organs. The drug vector's release and targeting system can reduce drug degradation and loss, lower side effects, and improve bioavailability. For example, polymer surfactants used as vectors can self-assemble to form various types of aggregates due to their unique amphiphilic structure, including micelles, microemulsions, gels, liquid crystals, and vesicles. These aggregates have the ability to encapsulate drug molecules and good membrane permeability, making them effective drug vectors.

[0123] The term "excipient" refers to a substance added to a drug formulation other than the active ingredient, and may also be called an additive. For example, adhesives, fillers, disintegrants, and lubricants in tablets, matrix components in semi-solid preparations, ointments, and creams, preservatives, antioxidants, flavoring agents, fragrances, cosolvents, emulsifiers, solubilizers, osmotic pressure adjusters, and coloring agents in liquid preparations, can all be called excipients.

[0124] The term "diluent," also known as a filler, is primarily used to increase the weight and volume of tablets. The addition of a diluent not only ensures a consistent volume, but also reduces the dosage deviation of the active ingredient and improves the compressibility of the drug. When a tablet contains an oily ingredient, an absorbent must be added to absorb the oily substance in order to maintain a "dry" state and facilitate tablet formulation. Examples include starch, lactose, inorganic calcium salts, and microcrystalline cellulose.

[0125] Reagents that provide basic conditions include organic bases and inorganic bases, the organic bases including, but not limited to, triethylamine, diethylamine, N-methylmorpholine, pyridine, hexahydropyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide, tetrabutylammonium fluoride, or potassium tert-butoxide, and the inorganic bases including, but not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, and potassium hydroxide, preferably N,N-diisopropylethylamine. [Brief explanation of the drawings]

[0126] [Figure 1] 1 shows the inhibitory effect of compounds according to the present disclosure on human acute lymphoblastic leukemia RS4;11 tumors subcutaneously transplanted into mice. [Figure 2] 1 shows the inhibition of body weight change caused by the compound according to the present disclosure in subcutaneously transplanted mice with the human acute lymphoblastic leukemia tumor RS4;11. DETAILED DESCRIPTION OF THE INVENTION

[0127] The present disclosure will be further described below in conjunction with examples, but these examples are not intended to limit the scope of the present disclosure.

[0128] Experimental methods for which specific conditions are not specified in the examples of this disclosure generally follow conventional conditions or conditions recommended by the manufacturers of raw materials or products. Reagents for which specific sources are not specified are commonly available commercially.

[0129] 1. Compound synthesis Example The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The NMR shifts (δ) are 10 -6The concentration is expressed in ppm (ppm). NMR measurements were performed using a Bruker AVANCE NEO 500M nuclear magnetic resonance spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as the measurement solvents, and tetramethylsilane (TMS) as the internal standard.

[0130] For MS measurements, liquid chromatograph mass spectrometers Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS (manufacturer: Agilent, MS model number: 6110 / 6120 Quadrupole MS), waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model number: waters ACQuity Qda Detector / waters SQ Detector), and THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model number: THERMO Q Exactive) were used.

[0131] High performance liquid chromatography (HPLC) analysis was performed using high performance liquid chromatographs Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489.

[0132] For chiral HPLC analysis, a high performance liquid chromatograph Agilent 1260 DAD was used.

[0133] For preparative high performance liquid chromatography, preparative chromatographs Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson GX-281 were used.

[0134] For chiral separation, a preparative chromatograph Shimadzu LC-20AP was used.

[0135] Combiflash Rf200 (TELEDYNE ISCO) was used as the CombiFlash high-speed preparative chromatograph.

[0136] Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used as silica gel plates for thin layer chromatography (TLC). The specifications for silica gel plates used in thin layer chromatography (TLC) are 0.15 mm to 0.2 mm, and the specifications for separating and purifying products by thin layer chromatography are 0.4 mm to 0.5 mm.

[0137] For silica gel column chromatography, 200-300 mesh silica gel manufactured by Yantai Huanghai Silica Gel was generally used as the vector.

[0138] Mean kinase inhibition rate and IC 50 The values ​​were measured using a microplate reader NovoStar (BMG, Germany).

[0139] Known starting materials according to the present disclosure may be synthesized by adopting or according to methods known in the art, or may be purchased commercially from commercial sources such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc.), and Darui Chemical. Unless otherwise stated in the examples, all reactions can be carried out in an argon or nitrogen atmosphere.

[0140] An argon or nitrogen atmosphere refers to an argon or nitrogen balloon with a volume of approximately 1 L connected to the reaction flask.

[0141] A hydrogen atmosphere refers to a hydrogen balloon with a volume of approximately 1 L attached to the reaction flask.

[0142] For the pressurized hydrogenation reaction, a Parr 3916EKX hydrogenator and a Seiran QL-500 hydrogen generator or an HC2-SS hydrogenator were used.

[0143] The hydrogenation reaction was usually carried out by repeating the procedure of evacuating and refilling with hydrogen three times.

[0144] A CEM Discover-S 908860 microwave reactor was used for the microwave reactions.

[0145] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0146] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0147] In the examples, thin layer chromatography (TLC) was used to monitor the progress of the reaction. The developing solvents used in the reaction, the eluent system of column chromatography for purifying the compounds, and the developing solvent system of thin layer chromatography comprised A: dichloromethane / methanol system and B: n-hexane / ethyl acetate system, and the volume ratio of the solvents may be adjusted according to the polarity of the compounds, or by adding small amounts of basic or acidic reagents such as triethylamine and acetic acid.

[0148] Example 2-1 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((R)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 1 [ka] [ka] Step 1 (R,Z)-tert-Butyl 2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)pyrrolidine-1-carboxylate 1c Methyl 2-(((benzyloxy)carbonyl)amino)-2-(dimethoxyphosphoryl)acetate 1b (1 g, 3.02 mmol, Shaoyuan) was dissolved in dichloromethane (10 mL) and cooled to 0 °C in an ice bath. 1,8-diazabicyclo[5.4.0]undec-7-ene (356 mg, 2.34 mmol, supplier: Shaoyuan) was added dropwise and the mixture was stirred at 0 °C for 20 minutes. Then, a 5 mL solution of tert-butyl (R)-2-formylpyrrolidine-1-carboxylate 1a (500 mg, 2.51 mmol, supplier: Yaoshi) in dichloromethane was added dropwise. After the addition was complete, the ice bath was removed, the mixture was warmed to room temperature, and the mixture was allowed to react for 48 hours. The reaction mixture was concentrated under reduced pressure, and the residue obtained using eluent system B was purified by silica gel column chromatography to give the title product 1c (830 mg, yield: 81.8%). MS m / z (ESI): 305.0 [M+H-100].

[0149] Step 2 (R)-2-((R)-2-amino-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylate tert-Butyl ester 1d 1c (520 mg, 1.28 mmol) was dissolved in isopropanol (10 mL), palladium on carbon (100 mg, 10% dry), and the mixture was purged with hydrogen gas three times and stirred at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated to give the title product 1d (300 mg, yield: 85.7%). The product was used in the next step without further purification. MS m / z (ESI): 273.1 [M+1].

[0150] Step 3 (R)-2-((R)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylate tert-Butyl ester 1e Crude product 1d (300 mg, 1.10 mmol) was dissolved in 1,4-dioxane (4 mL), water (1 mL), sodium bicarbonate (278 mg, 3.31 mmol), and 9-fluorenylmethyl chloroformate (285 mg, 1.10 mmol, Shaoyuan) were added, and the mixture was stirred at room temperature for 1.5 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system B to give the title product 1e (396 mg, yield: 72.7%). MS m / z (ESI): 395.1 [M+H-100].

[0151] Step 4 (R)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxypropyl)pyrrolidine-1-carboxylate tert-Butyl ester 1f 1e (200 mg, 0.40 mmol) was dissolved in tetrahydrofuran (10 mL), ethanol (10 mL) was added, and sodium borohydride (92 mg, 2.43 mmol) and potassium chloride (112 mg, 2.64 mmol) were added sequentially. The reaction mixture was stirred at 28 °C for 2.5 h. The reaction mixture was cooled in an ice bath and quenched by the dropwise addition of saturated ammonium chloride solution (10 mL). Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 1f (144 mg, 76.3%). MS m / z (ESI): 367.1 [M+H-100].

[0152] Step 5 (R)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate tert-butyl 1g 1f (144 mg, 0.31 mmol) was dissolved in toluene (5 mL), and diphenyl disulfide (225 mg, 1.03 mmol, adamas) and tributylphosphine (208 mg, 1.03 mmol) were added. The reaction mixture was purged with nitrogen gas three times, then heated to 80°C and stirred for 12 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 1g (168 mg, 97.4%). MS m / z (ESI): 459.1 [M+H-100].

[0153] Step 6 (R)-2-((R)-2-amino-3-(phenylthio)propyl)pyrrolidine-1-carboxylate tert-butyl ester 1h 1g (168 mg, 0.30 mmol) was dissolved in dichloromethane (2 mL), diethylamine (2 mL) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to remove the organic solvent, and the mixture was concentrated under reduced pressure with toluene (5 mL) to give the crude title product 1h (101 mg), which was carried on to the next step without further purification. MS m / z (ESI): 337.1 [M+1].

[0154] Step 7 (R)-2-((R)-3-(phenylthio)-2-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)propyl)pyrrolidine-1-carboxylate tert-Butyl ester 1j The crude product 1h (101 mg, 0.30 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 4-fluoro-3-(trifluoromethylsulfonyl)benzenesulfonamide 1i (102 mg, 0.33 mmol) and N,N-diisopropylethylamine (388 mg, 3.00 mmol) were added. The reaction mixture was purged with nitrogen gas three times, then heated to 50 °C and stirred for 8 h. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 1j (129 mg, 68.9%). MS m / z (ESI): 524.0 [M+H-100].

[0155] Step 8 tert-Butyl (R)-2-((R)-2-((4-(N-(4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate 1l 1j (22 mg, 0.035 mmol) was dissolved in dichloromethane (3 mL) and 1k (16 mg, 0.038 mmol, synthesized by reference to intermediate 40 on page 79 of the patent specification "CN103153954 B") was added. 4-Dimethylaminopyridine (8.6 mg, 0.070 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (20.3 mg, 0.106 mmol) were then added. After addition was complete, the mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. 1k (11 mg, 0.026 mmol) was then added and the mixture was stirred overnight at room temperature. The reaction mixture was washed sequentially with water (5 mL) and saturated sodium chloride solution (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using developing solvent system A to give the title product 1l (28 mg, yield: 77.2%). MS m / z (ESI): 1027.0 [M+1].

[0156] Step 9 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((R)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 1 1L (14 mg, 0.014 mmol) was dissolved in dichloromethane (0.2 mL), and 4 M hydrogen chloride in dioxane (0.5 mL) was added. The mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure to remove the organic solvent. The residue was slurried with ether and filtered. The resulting solid was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 1 (5 mg, yield: 39.6%). MS m / z (ESI): 927.2 [M+1]. 1H NMR (500 MHz, CD3OD) δ 8.25 (s, 1H), 8.06 (d, 1H), 7.82 (d, 2H), 7.62 (d, 1H), 7.48-7.31 (m, 8H), 7.26 (t, 2H), 7.20 (t, 2H), 6.82 (d, 2H), 6.74 (d, 1H), 4.44 (d, 1H), 4.00-3.95 (m, 1H), 3.78 (d, 1H), 3.61 (d, 1H), 3.52-3.45 (m, 2H), 3.26-3.17 (m, 2H), 2.68 (t, 2H), 2.53 (t, 2H), 2.27-1.92 (m, 6H), 1.81-1.59 (m, 4H), 1.27-1.21 (m, 1H), 1.18-1.12 (m, 1H), 1.02-0.90 (m, 2H).

[0157] Example 2-2 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((R)-1-(2-hydroxyethyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 2 [ka] [ka] 1 (13 mg, 0.014 mmol) was placed in a reaction flask, and acetonitrile (5 mL) was added. 2-Bromoethanol (26 mg, 0.210 mmol, adamas) and triethylamine (11 mg, 0.112 mmol) were then added. The reaction mixture was purged with nitrogen gas three times, then heated to 70 °C and stirred for 7 h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 2 (3 mg, yield: 22.0%). MS m / z (ESI): 971.2 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.26 (s, 1H), 8.03 (d, 1H), 7.80 (d, 2H), 7.62 (d, 1H), 7.46-7.36 (m, 5H), 7.35-7.27 (m, 5H), 7.24 (d, 1H), 7.18 (d, 1H), 6.81 (d, 2H), 6.63 (d, 1H), 4.42 (d, 1H), 3.91-3.77 (m, 4H),3.63 (d, 2H), 3.25-3.16 (m, 2H), 2.67 (t, 2H), 2.54 (t, 2H), 2.36-2.29 (m, 2H), 2.23-2.13 (m, 3H), 2.09-1.97 (m, 4H),1.81-1.71 (m, 2H), 1.65-1.59 (m, 2H), 1.27-1.21 (m, 1H), 1.17-1.11 (m, 1H), 1.02-0.90 (m, 2H).

[0158] Example 2-3 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((R)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 3 [ka] [ka]

[0159] Step 1 (R)-4-(4-((4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)benzoate 3b Ethyl (R)-4-(4-((4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)benzoate 3a (60 mg, 0.133 mmol, synthesized with reference to the method according to intermediate 11 on page 63 of the specification of patent "CN103153954 B") was dissolved in 2 mL of a mixed solvent of N,N-dimethylformamide and tetrahydrofuran (V:V=1:1). The mixture was purged with nitrogen gas three times, and iodomethane (190 mg, 1.34 mmol) and 60% sodium hydride (11 mg, 0.275 mmol) were added in that order, followed by stirring at room temperature for 3 hours. The reaction mixture was cooled in an ice bath, saturated ammonium chloride solution (5 mL) was added to quench the reaction, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, the desiccant was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 3b (57 mg, 92.1%). MS m / z (ESI): 464.1 [M+1].

[0160] Step 2 (R)-4-(4-((4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)benzoic acid 3c 3b (57 mg, 0.122 mmol) was dissolved in 3 mL of a mixed solvent of tetrahydrofuran, methanol, and water (V:V:V = 4:1:1), potassium hydroxide monohydrate (26 mg, 0.620 mmol) was added, and the reaction mixture was heated to 50 °C and stirred for 16 hours. The reaction mixture was concentrated under reduced pressure, and the pH was adjusted to 2-3 with 1 M hydrochloric acid. The filtered cake was dissolved in 10 mL of dichloromethane, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title product 3c (53 mg, yield: 98.9%). MS m / z (ESI): 436.1 [M+1].

[0161] Step 3 tert-Butyl (R)-2-((R)-2-((4-(N-(4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate 3d 1j (32 mg, 0.051 mmol) was dissolved in dichloromethane (5 mL) and 3c (24 mg, 0.055 mmol) was added. 4-Dimethylaminopyridine (13 mg, 0.106 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (30 mg, 0.156 mmol) were then added. After the addition was complete, the mixture was stirred at room temperature under a nitrogen atmosphere for 4 h. After adding 3c (22 mg, 0.051 mmol), the mixture was stirred at room temperature overnight. The reaction mixture was washed successively with water (10 mL) and saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using developing solvent system A to give the title product 3d (53 mg, yield: 99.6%). MS m / z (ESI): 1041.1 [M+1].

[0162] Step 4 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((R)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 3 3d (30 mg, 0.028 mmol) was dissolved in dichloromethane (0.4 mL), and 4 M hydrogen chloride in dioxane (1.2 mL) was added. The mixture was stirred at room temperature for 40 minutes. The reaction mixture was concentrated under reduced pressure to remove the organic solvent. The residue was slurried with ether and filtered. The resulting solid was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 3 (10 mg, yield: 36.9%). MS m / z (ESI): 941.1 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.26 (s, 1H), 8.04 (dd, 1H), 7.81 (d, 2H), 7.50 (d, 1H), 7.47-7.41 (m, 3H), 7.40-7.32 (m, 3H), 7.30-7.23 (m, 4H), 7.22-7.16 (m, 2H),6.81 (d, 2H), 6.72 (d, 1H), 4.06 (d, 1H), 3.99-3.93 (m, 1H), 3.77 (d, 1H), 3.66 (d, 1H), 3.55-3.49 (m, 1H), 3.23 (d, 2H), 3.17 (s, 3H), 2.61 (t, 2H), 2.51 (t, 2H), 2.25-1.93 (m, 6H), 1.72-1.58 (m, 4H), 1.27-1.23 (m, 1H), 1.20-1.11 (m, 2H), 0.91 (t, 1H).

[0163] Examples 2-4 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(methoxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((R)-1-(2-hydroxyethyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 4 [ka] [ka] 3 (15 mg, 0.015 mmol) was placed in a reaction flask, and acetonitrile (5 mL) was added. 2-Bromoethanol (29 mg, 0.232 mmol) and triethylamine (16 mg, 0.158 mmol) were then added. The reaction mixture was purged with nitrogen gas three times, then heated to 70 °C and stirred for 16 h. The reaction mixture was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 4 (2.5 mg, yield: 16.5%). MS m / z (ESI): 985.1 [M+1]. 1H NMR (500 MHz, CD3OD) δ 8.27 (s, 1H), 8.03 (dd, 1H), 7.81 (d, 2H), 7.51 (d, 1H), 7.47-7.39 (m, 5H), 7.35 (t, 1H), 7.32-7.21 (m, 5H), 7.18 (d, 1H), 6.82 (d, 2H), 6.63 (d, 1H), 4.06 (d, 1H), 3.90-3.75 (m, 4H), 3.69-3.63 (m, 1H), 3.27-3.20 (m, 2H), 3.17 (s, 3H), 2.62 (t, 2H), 2.52 (t, 2H), 2.36-2.28 (m, 2H), 2.23-2.13 (m, 3H), 2.09-1.94 (m, 4H), 1.78-1.58 (m, 4H), 1.28-1.24 (m, 1H), 1.21-1.13 (m, 2H), 0.91(t, 1H).

[0164] Examples 2-5 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((R)-1-(3-hydroxypropyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 5 [ka] [ka] 1 (10 mg, 0.011 mmol) was placed in a reaction flask, and acetonitrile (3 mL) was added. 3-Bromopropanol (15 mg, 0.108 mmol, supplied by Adamas) and triethylamine (11 mg, 0.108 mmol) were then added. The reaction mixture was purged with nitrogen gas three times, then heated to 70 °C and stirred for 16 h. The reaction mixture was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 5 (2.5 mg, yield: 23.5%). MS m / z (ESI): 985.1 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.26 (s, 1H), 8.03 (d, 1H), 7.81 (d, 2H), 7.61 (d, 1H), 7.46-7.39 (m, 5H), 7.35-7.26 (m, 5H), 7.22 (t, 1H), 7.18 (d, 1H), 6.81 (d, 2H), 6.62 (d, 1H), 4.43 (d, 1H), 3.89-3.84 (m, 1H), 3.79 (d, 1H), 3.67 (t, 2H), 3.63 (d, 2H), 3.25-3.15 (m, 2H), 2.67 (t, 2H), 2.53 (t, 2H), 2.32-2.26 (m, 2H), 2.19 (t, 2H), 2.15-2.11 (m, 1H), 2.08-1.95 (m, 4H), 1.92-1.84 (m, 2H), 1.79-1.67 (m, 2H), 1.65-1.58 (m, 2H), 1.26-1.22 (m, 1H), 1.16-1.12 (m, 1H), 1.02-0.96 (m, 1H), 0.91 (t, 1H).

[0165] Examples 2-6 4-(4-((S)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-4-((2-hydroxyethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 6 [ka] [ka]

[0166] Step 1 tert-Butyl 4-(hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidine-1-carboxylate 6b 2-Bromo-4'-(trifluoromethyl)-1,1'-biphenyl 6a (3.3 g, 10.95 mmol, synthesized by the method disclosed in Example 53 on page 53 of Patent "US2004 / 171833, 2004, A1") was placed in a flask, tetrahydrofuran (50 mL) was added, and the temperature was lowered to -78 ° C under a nitrogen atmosphere. n-Butyllithium (1.40 g, 21.92 mmol) was added dropwise to the reaction flask, and the internal temperature was maintained below -65 ° C. After the addition was completed, the mixture was stirred at -78 ° C for 40 minutes. A solution of tert-butyl formylpiperidine-1-carboxylate (4.67 g, 21.92 mmol) in tetrahydrofuran (20 mL) was added, and after the addition was completed, the mixture was heated to 0 ° C. and reacted for 3 hours. The mixture was quenched by adding saturated aqueous ammonium chloride solution dropwise, concentrated under reduced pressure, and extracted with ethyl acetate (40 mL × 3). The organic phases were combined and washed with water (40 The resulting mixture was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 6b (5.5 g, yield: >100%). MS m / z (ESI): 434.1 [M-1].

[0167] Step 2 Piperidin-4-yl(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methanol 6c 6b (5.5 g, 12.63 mmol) was placed in a reaction flask, and 4 M dioxane hydrochloric acid solution (20 mL) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react for 2 hours with stirring. The reaction mixture was concentrated under reduced pressure, and toluene (10 mL) was added to the residue. The mixture was then concentrated under reduced pressure and dried by suction using an oil pump to obtain the crude title product 6c (4.23 g, yield: 100%). The product was directly used in the next step without further purification. MS m / z (ESI): 336.0 [M+1].

[0168] Step 3 (R)-4-(4-(hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzoate 6d Ethyl (S)-4-(4-(hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzoate 6e Crude product 6c (270 mg, 0.805 mmol) was placed in a reaction flask, and dimethyl sulfoxide (3 mL) was added. Ethyl p-fluorobenzoate (270 mg, 1.61 mmol, Shaoyuan) and N,N-diisopropylethylamine (1.04 g, 8.05 mmol) were added. After the addition was complete, the mixture was heated to 120 °C under a nitrogen atmosphere and reacted for 24 h. 5 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (4 mL × 5). The combined organic phases were washed with water (4 mL), washed with saturated brine (4 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to obtain 360 mg of a mixture, which was then sent to chiral preparative separation (preparative conditions: column: CHIRALPAK IG, 0.50 cm ID * 25 cm L, 0.1 μm, mobile phase: MeOH = 100%, detection wavelength: UV 214 nm, temperature: 38 ° C). Two components were obtained: Peak 1 with a relatively short retention time was 6d (73 mg), and Peak 2 with a relatively long retention time was 6e (72 mg). MS m / z (ESI): 484.1 [M+1]. Peak 1: Chiral HPLC analysis: retention time 4.008 minutes, chiral purity: 100% (column: CHIRALPAK IG 150*4.6 mm, 5 μm (with guard column), mobile phase: n-hexane / ethanol = 70 / 30 (v / v)). Peak 2: Chiral HPLC analysis: retention time 6.289 minutes, chiral purity: 100% (column: CHIRALPAK IG 150*4.6 mm, 5 μm (with guard column), mobile phase: n-hexane / ethanol = 70 / 30 (v / v)).

[0169] Step 4 (R)-4-(4-(hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzoic acid 6f 6e (70 mg, 0.145 mmol) was placed in a flask, and tetrahydrofuran (2 mL), water (0.5 mL), and methanol (0.5 mL) were added. Potassium hydroxide monohydrate (30.3 mg, 0.723 mmol) was then added, and the mixture was heated to 50°C under a nitrogen atmosphere and reacted for 16 hours. The reaction mixture was concentrated under reduced pressure, and water (5 mL) was added to the residue. The mixture was cooled in an ice bath, adjusted to pH 4 with 1N hydrochloric acid solution, and filtered. The filter cake was rinsed with n-hexane (5 mL) and dried by suction using an oil pump to give the title product 6f (63 mg, yield: 95.54%). MS m / z (ESI): 456.0 [M+1].

[0170] Step 5 N-((4-(((R)-4-((2-((tert-butyldimethylsilyl)oxy)ethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)-4-(4-((S)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzamide 6h 6g (30 mg, 0.040 mmol, synthesized by referring to the method of intermediate 67 on page 90 of the specification of patent "CN103153954 B") was dissolved in dichloromethane (3 mL), 6f (25 mg, 0.055 mmol) was added, and then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (17.5 mg, 0.091 mmol) and 4-dimethylaminopyridine (11.2 mg, 0.091 mmol) were added in that order. After the addition was completed, the mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. 6f (16.6 mg, 0.037 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (10.5 mg, 0.055 mmol), and 4-dimethylaminopyridine (6.7 mg, 0.055 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction mixture was washed with water (5 mL), saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using elution system A to give the title product 6h (40 mg, yield: 79.9%). MS m / z (ESI): 1093.4 [M+1].

[0171] Step 6 4-(4-((S)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-4-((2-hydroxyethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 6 6h (40 mg, 0.036 mmol) was dissolved in tetrahydrofuran (2 mL), tetrabutylammonium fluoride (16.4 mg, 0.073 mmol) was added, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (8 mL) and washed with saturated ammonium chloride solution (10 mL × 2). The organic phase was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (8 mL), washed with saturated ammonium chloride solution (10 mL × 6), washed with water (10 mL), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (HPLC) (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 6 (17 mg, yield: 47.5%). MS m / z (ESI): 979.2 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.28 (d, 1H), 8.09-8.03 (m, 1H), 7.80 (d, 2H), 7.74 (d, 2H), 7.66 (d, 1H), 7.55 (d, 2H), 7.48 (t, 1H), 7.42-7.34 (m, 2H), 7.29-7.16 (m, 3H), 6.81 (dd, 2H), 4.41 (d, 1H), 4.02-3.97 (m, 1H), 3.84-3.75 (m, 2H), 3.64 (d, 1H), 3.37 (s, 2H), 3.30-3.18 (m, 3H), 3.12-3.06 (m, 2H), 2.75 (s, 2H), 2.72-2.63 (m, 1H), 2.59-2.50 (m, 1H), 2.29-2.19 (m, 2H), 2.08-2.02 (m, 2H), 1.82-1.72 (m, 1H), 1.66-1.58 (m, 1H), 1.39-1.31 (m, 4H), 1.27-1.12 (m, 2H), 1.03-0.89 (m, 2H).

[0172] Examples 2-7 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((3R)-1-((2-hydroxyethyl)(methyl)amino)-4-(phenylthio)pentan-3-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 7 [ka] [ka] [ka]

[0173] Step 1 (R)-3-(((benzyloxy)carbonyl)amino)-4-oxobutyric acid methyl ester 7b Oxalyl chloride (772 mg, 6.08 mmol) was dissolved in dichloromethane (5 mL), the mixture was purged with nitrogen gas three times, and the mixture was cooled to -78°C. A dichloromethane solution (5 mL) of dimethyl sulfoxide (950 mg, 12.16 mmol) was added dropwise. After the addition was completed, the mixture was stirred at -78°C for 30 minutes. A dichloromethane solution (5 mL) of (R)-3-(((benzyloxy)carbonyl)amino)-4-hydroxybutyrate methyl 7a (650 mg, 2.43 mmol, synthesized with reference to the method according to intermediate 4 on page 59 of the specification of patent "CN103153954 B") was added dropwise. After the addition was completed, the mixture was stirred at -78°C for 1 hour. N,N-diisopropylethylamine (3.14 g, 24.29 mmol) was added dropwise. After the addition was complete, the reaction mixture was warmed to -60°C and stirred for 30 minutes, then warmed to 0°C and stirred for 30 minutes. The reaction was quenched by the addition of ice-cold 1 M hydrochloric acid (20 mL) at 0°C, and the layers were separated. The organic phase was washed with pH 7 phosphate buffer solution (30 mL) and saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7b (540 mg, yield: 83.7%). MS m / z (ESI): 266.2 [M+1].

[0174] Step 2 (3R)-3-(((benzyloxy)carbonyl)amino)-4-hydroxypentanoic acid methyl ester 7c 7b (490 mg, 1.95 mmol) was dissolved in ether (15 mL), purged with nitrogen gas three times, cooled to -78 °C, and a 1 M solution of methylmagnesium bromide in tetrahydrofuran (2 mL, 2.03 mmol) was added dropwise. After the addition was complete, the mixture was allowed to warm to 0 °C and stirred for 3 h. The reaction was quenched by the addition of saturated ammonium chloride solution (20 mL) at 0 °C, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7c (188 mg, yield: 36.2%). MS m / z (ESI): 282.1 [M+1].

[0175] Step 3 (3R)-3-(((benzyloxy)carbonyl)amino)-4-((methylsulfonyl)oxy)pentanoic acid methyl ester 7d 7c (220 mg, 0.782 mmol) was dissolved in dichloromethane (8 mL), pyridine (93 mg, 1.176 mmol) and 4-dimethylaminopyridine (9.5 mg, 0.078 mmol) were added, and the mixture was purged with nitrogen three times. The mixture was cooled to 0 °C, and methanesulfonic anhydride (164 mg, 0.941 mmol) was slowly added. After the addition was complete, the reaction mixture was warmed to room temperature and stirred for 1.5 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (8 mL x 3). The combined organic phase was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7d (216 mg, yield: 76.8%). MS m / z (ESI): 360.0 [M+1].

[0176] Step 4 (3R)-3-(((benzyloxy)carbonyl)amino)-4-(phenylthio)pentanoic acid methyl ester 7e 7d (216 mg, 0.601 mmol) was dissolved in toluene (8 mL), sodium thiophenol (397 mg, 3.004 mmol) was added, and the mixture was purged with nitrogen gas three times. The mixture was heated to 105°C and stirred for 16 hours. After cooling to room temperature, water (20 mL) was added and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7e (120 mg, yield: 53.5%). MS m / z (ESI): 374.1 [M+1].

[0177] Step 5 (3R)-4-(phenylthio)-3-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)pentanoic acid methyl ester 7f 7e (112 mg, 0.300 mmol) was dissolved in tetrahydrofuran (1.5 mL), trifluoroacetic acid (5 mL) was added, and the reaction mixture was heated to 72°C and stirred for 1.5 hours. The mixture was concentrated under reduced pressure, concentrated under reduced pressure with dichloromethane (20 mL), and dried in vacuo for 2 hours. The residue was dissolved in N,N-dimethylformamide (4 mL), and 1i (93 mg, 0.302 mmol) and N,N-diisopropylethylamine (388 mg, 3.00 mmol) were added. The reaction mixture was purged with nitrogen gas three times, then heated to 50°C and stirred for 24 hours. Water (20 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, the desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7f (119 mg, 75.3%). MS m / z (ESI): 524.9 [M-1].

[0178] Step 6 (3R)-4-(phenylthio)-3-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)pentanoic acid 7g 7f (119 mg, 0.226 mmol) was dissolved in 5 mL of a mixed solvent of tetrahydrofuran, methanol, and water (V:V:V = 3:1:1). Potassium hydroxide monohydrate (29 mg, 0.691 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and the pH was adjusted to 3-4 with 1 M hydrochloric acid. The mixture was extracted with dichloromethane (8 mL x 3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and the desiccant was removed by filtration. The filtrate was concentrated under reduced pressure to give the title product 7g (115 mg, yield: 99.3%). MS m / z (ESI): 511.0 [M-1].

[0179] Step 7 (3R)-N-(2-((tert-butyldimethylsilyl)oxy)ethyl)-N-methyl-4-(phenylthio)-3-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)pentanamide 7h 7g (115 mg, 0.224 mmol) was dissolved in tetrahydrofuran (3 mL), and 2-((tert-butyldimethylsilyl)oxy)-N-methylethan-1-amine (43 mg, 0.227 mmol, Shaoyuan), 3-(diethoxy-o-acyloxy)-1,2,3-benzotriazin-4-one (134 mg, 0.448 mmol), and N,N-diisopropylethylamine (58 mg, 0.448 mmol) were added in that order. After the addition was complete, the reaction was carried out under nitrogen atmosphere at room temperature with stirring for 3 hours. The reaction mixture was concentrated, and the residue was dissolved in ethyl acetate (10 mL). The solution was washed successively with saturated aqueous ammonium chloride (20 mL), saturated sodium bicarbonate (20 mL), and saturated sodium chloride (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system B to give the title product 7h (107 mg, yield: 69.7%). MS m / z (ESI): 684.0 [M+1].

[0180] Step 8 4-(((3R)-1-((2-((tert-butyldimethylsilyl)oxy)ethyl)(methyl)amino)-4-(phenylthio)pentan-3-yl)amino)-3-((trifluoromethyl)sulfonyl)benzenesulfonamide 7i 7h (107 mg, 0.156 mmol) was placed in a reaction flask, and 1 M borane-tetrahydrofuran complex solution (5 mL) was added. After the addition was complete, the mixture was heated to 60°C and stirred for 24 hours. The reaction mixture was cooled to 0°C and quenched by the dropwise addition of 4 mL of methanol. The mixture was concentrated under reduced pressure. To the residue was added 7 M ammonia in methanol (10 mL). The mixture was sealed and stirred at room temperature for 48 hours. The mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 7i (10 mg, yield: 9.5%). MS m / z (ESI): 670.1 [M+1].

[0181] Step 9 (R)-4-(4-(hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzoic acid 7j 6d (70 mg, 0.145 mmol) was placed in a reaction flask, and tetrahydrofuran (2 mL), water (0.5 mL), and methanol (0.5 mL) were added. Potassium hydroxide monohydrate (30 mg, 0.72 mmol) was then added, and the mixture was heated to 50°C under a nitrogen atmosphere and reacted for 16 hours. The reaction mixture was concentrated under reduced pressure, and water (5 mL) was added to the residue. The mixture was cooled in an ice bath, adjusted to pH 4 with 1 M hydrochloric acid solution, and filtered. The filter cake was rinsed with n-hexane (5 mL) and dried by oil pump suction to give the title product 7j (63 mg, yield: 95.5%). MS m / z (ESI): 456.0 [M+1].

[0182] Step 10 N-((4-(((3R)-1-((2-((tert-butyldimethylsilyl)oxy)ethyl)(methyl)amino)-4-(phenylthio)pentan-3-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)-4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzamide 7k 7i (16 mg, 0.024 mmol) was dissolved in dichloromethane (4 mL), 7j (13 mg, 0.028 mmol) was added, and 4-dimethylaminopyridine (6 mg, 0.049 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (14 mg, 0.073 mmol) were added. The reaction mixture was purged with nitrogen gas three times and stirred at room temperature for 4 h. After the addition of 7j (13 mg, 0.028 mmol), the mixture was stirred at room temperature for 16 h. The reaction mixture was washed successively with water (8 mL) and saturated sodium chloride solution (8 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by thin-layer chromatography using developing solvent system A to give the title product 7k (14 mg, yield: 52.9%). MS m / z (ESI): 1107.2 [M+1].

[0183] Step 11 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((3R)-1-((2-hydroxyethyl)(methyl)amino)-4-(phenylthio)pentan-3-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 7 7k (14 mg, 0.013 mmol) was dissolved in tetrahydrofuran (3 mL), tetrabutylammonium fluoride (6 mg, 0.027 mmol) was added, and the mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (5 mL) and washed with saturated ammonium chloride solution (5 mL × 2). The organic phase was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (8 mL) and washed sequentially with saturated ammonium chloride solution (5 mL × 6), water (10 mL), and saturated sodium chloride solution (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (HPLC) (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 7 (4.5 mg, yield: 35.8%). MS m / z (ESI): 993.2 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.33 (s, 1H), 8.02 (d, 1H), 7.76 (dd, 4H), 7.65 (d, 1H), 7.56 (dd, 4H), 7.48 (t, 1H), 7.44-7.33 (m, 4H), 7.21 (d, 1H), 6.81 (d, 2H), 6.60 (d, 1H), 4.41 (d, 1H), 3.97-3.92 (m, 1H), 3.84-3.72 (m, 3H), 3.69-3.56 (m, 3H), 2.76-2.62 (m, 4H), 2.55 (t, 2H), 2.32-2.17 (m, 3H), 2.10-1.87 (m, 4H), 1.82-1.72 (m, 2H), 1.64-1.58 (m, 2H), 1.27-1.22 (m, 1H), 1.21-1.12 (m, 2H), 1.03-0.96 (m, 1H), 0.92 (t, 1H).

[0184] Examples 2-8 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((S)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 8 [ka] [ka]

[0185] Step 1 (S)-tert-Butyl 2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxoprop-1-en-1-yl)pyrrolidine-1-carboxylate 8b 1b (2.49 g, 7.53 mmol, Shaoyuan) was dissolved in tetrahydrofuran (20 mL), cooled to 0°C in an ice bath, N,N-lithium diisopropylamide (661 mg, 6.17 mmol) was added dropwise, and the reaction was allowed to proceed with stirring at 0°C for 1 hour. The reaction mixture was then cooled to -78°C, and 20 mL of a tetrahydrofuran solution of (S)-2-formylpyrrolidine-1-carboxylate tert-butyl 8a (1.00 g, 5.02 mmol, Yakushi) was added dropwise. After the addition was complete, the dry ice acetone bath was removed, the mixture was warmed to room temperature, and the reaction was continued for 12 hours. The reaction mixture was quenched by adding saturated ammonium chloride (20 mL) and extracted with ethyl acetate (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title product 8b (1.90 g, yield: 94.1%). MS m / z (ESI):305.0 [M+H-100].

[0186] Step 2 (2S)-2-(2-amino-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylate tert-Butyl ester 8c 8b (1.90 g, 4.7 mmol) was dissolved in isopropanol (50 mL), palladium on carbon (380 mg, 10% dry content) was added, and the mixture was purged with hydrogen gas three times and stirred at room temperature for 8 hours. The reaction mixture was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate (20 mL) and methanol (20 mL). The filtrate was concentrated to give the crude title product 8c (1.28 g), which was used directly in the next step without further purification.

[0187] Step 3 (S)-2-((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylate tert-Butyl ester 8d (S)-2-((R)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylate tert-Butyl ester 8e Crude product 8c (1.28 g, 4.7 mmol) was dissolved in 1,4-dioxane (40 mL), water (10 mL), sodium bicarbonate (1.17 g, 14.1 mmol), and 9-fluorenylmethyl chloroformate (1.21 g, 4.7 mmol) were added, and the mixture was stirred at room temperature for 3 h. The mixture was quenched with water (30 mL), extracted with ethyl acetate (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography using eluent system A to give the title products 8d (1.03 g, 44.2%, peak 1) and 8e (513 mg, 22.1%, peak 2). MS m / z (ESI): 495.2 [M+1]. Peak 1: UPLC analysis: retention time 2.77 min (Column: ACQUITY UPLC BEHC18 50*2.1 mm, 1.7 μm, Mobile phase: Gradient acetonitrile / water / formic acid = 10 / 90 / 0.1 (v / v / v) to 95 / 5 / 0.1 (v / v / v) rinse). Peak 2: UPLC analysis: retention time 2.69 min (Column: ACQUITY UPLC BEHC18 50*2.1 mm, 1.7 μm, Mobile phase: Gradient acetonitrile / water / formic acid = 50 / 50 / 0.1 (v / v / v) to 95 / 5 / 0.1 (v / v / v) rinse).

[0188] Step 4 (S)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxypropyl)pyrrolidine-1-carboxylate tert-Butyl ester 8f 8e (169 mg, 0.34 mmol) was dissolved in tetrahydrofuran (8 mL), ethanol (8 mL), sodium borohydride (77 mg, 2.05 mmol), and potassium chloride (94 mg, 2.22 mmol) were added sequentially, and the mixture was stirred at 28 °C for 2.5 h. The reaction was quenched by the dropwise addition of saturated ammonium chloride (5 mL). The solvent was concentrated to dryness, and the residue was dissolved in ethyl acetate (10 mL), washed with water (10 mL), and then washed with saturated sodium chloride (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system A to give the title product 8f (125 mg, 78.4%). MS m / z (ESI): 467.2 [M+1].

[0189] Step 5 (S)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate tert-butyl ester 8g 8f (120 mg, 0.26 mmol) was dissolved in toluene (5 mL), diphenyl disulfide (168 mg, 0.77 mmol) and tributylphosphine (156 mg, 0.77 mmol) were added, and the reaction mixture was heated to 80°C under a nitrogen atmosphere and stirred for 12 h. The reaction mixture was concentrated to dryness, and the resulting residue was purified by silica gel column chromatography using eluent system A to give the title product 8g (135 mg, 93.9%). MS m / z (ESI): 559.2 [M+1].

[0190] Step 6 (S)-2-((R)-2-amino-3-(phenylthio)propyl)pyrrolidine-1-carboxylate tert-butyl ester 8h 8g (135 mg, 0.24 mmol) was dissolved in dichloromethane (4 mL), diethylamine (4 mL) was added, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to remove the organic solvent, affording the title product 8h (81.6 mg), which was directly used in the next step without further purification.

[0191] Step 7 (S)-2-((R)-4-(phenylthio)-3-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)butyl)pyrrolidine-1-carboxylate tert-Butyl ester 8i Crude product 8h (81.6 mg, 0.24 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 1i (82 mg, 0.27 mmol) and N,N-diisopropylethylamine (314 mg, 2.4 mmol) were added. The reaction mixture was heated to 50 °C under a nitrogen atmosphere and stirred for 12 h. Water (10 mL) was added to quench the reaction, followed by extraction with ethyl acetate (5 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 8i (105 mg, yield: 69.3%). MS m / z (ESI): 622.1 [M-1].

[0192] Step 8 tert-Butyl (S)-2-((R)-2-((4-(N-(4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate 8j 8i (30 mg, 0.048 mmol) was dissolved in dichloromethane (3 mL), and 1k (24 mg, 0.058 mmol) was added. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19 mg, 0.096 mmol) and 4-dimethylaminopyridine (12 mg, 0.096 mmol) were then added sequentially. After the addition was complete, the mixture was stirred at room temperature under a nitrogen atmosphere for 4 h. 1k (12 mg, 0.038 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19 mg, 0.096 mmol), and 4-dimethylaminopyridine (12 mg, 0.096 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction mixture was washed with water (5 mL), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using developing solvent system A to give the title product 8j (30 mg, yield: 60.7%). MS m / z (ESI): 1027.2 [M+1].

[0193] Step 9 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((S)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 8 8j (30 mg, 0.029 mmol) was dissolved in dichloromethane (2 mL), 4 M hydrogen chloride in 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 20 minutes. The reaction mixture was then concentrated under reduced pressure to remove the organic solvent. The residue was dissolved in dichloromethane and concentrated under reduced pressure to dryness. This was repeated three times. The resulting residue was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 8 (25 mg, yield: 92.3%). MS m / z (ESI): 927.1 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.24 (d, 1H), 8.05-8.01 (m, 1H), 7.82 (d, 2H), 7.60 (d, 1H), 7.45-7.39 (m, 3H), 7.37 (d, 2H), 7.34-7.28 (m, 3H), 7.23 (t, 2H), 7.20-7.14 (m, 2H), 6.80 (d, 2H), 6.72 (d, 1H), 4.42 (d, 1H), 3.97 (br, 1H), 3.84-3.74 (m, 1H), 3.27-3.21 (m, 2H), 3.19-3.07 (m, 3H), 3.06-2.97 (m, 1H), 2.69-2.61 (m, 1H), 2.55-2.48 (m, 1H), 2.16-2.00 (m, 5H), 1.98-1.89 (m, 2H), 1.88-1.80 (m, 1H), 1.78-1.69 (m, 1H), 1.63-1.57 (m, 2H), 1.54-1.49 (m, 1H), 1.26-1.20 (m, 1H), 1.15-1.04 (m, 1H), 1.01-0.94 (m, 1H).

[0194] Examples 2-9 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(2-hydroxyethyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 9 [ka] [ka] 8 (15 mg, 0.016 mmol) was dissolved in acetonitrile (2 mL), triethylamine (16 mg, 0.16 mmol) and bromoethanol (20 mg, 0.16 mmol) were added, and the reaction mixture was heated to 70 °C under a nitrogen atmosphere for 12 h. The mixture was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile), and the corresponding components were collected and concentrated under reduced pressure to give the title product 9 (10 mg, yield: 63.4%). MS m / z (ESI): 971.2 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.25 (d, 1H), 8.06 (dd, 1H), 7.80 (d, 2H), 7.61 (dd, 1H), 7.49-7.35 (m, 5H), 7.35-7.28 (m, 3H), 7.24 (dd, 2H), 7.20-7.14 (m, 2H), 6.79 (t, 3H), 4.42 (d, 1H), 4.02 (t, 1H), 3.84-3.70 (m, 3H), 3.61 (d, 2H), 3.28-3.14 (m, 2H), 3.06 (br, 1H), 2.96 (br, 1H), 2.71-2.59 (m, 1H), 2.56-2.45 (m, 1H), 2.45-2.35 (m, 1H), 2.32-2.22 (m, 1H), 2.10-1.97 (m, 4H), 1.96-1.86 (m, 1H), 1.85-1.69 (m, 2H), 1.60 (t, 2H), 1.26-1.18 (m, 1H), 1.06-1.08 (m, 1H), 1.01-0.92 (m, 1H), 0.92-1.86 (m, 1H).

[0195] Example 2-10 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-4-((2-hydroxyethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 10 [ka] [ka]

[0196] Step 1 N-((4-(((R)-4-((2-((tert-butyldimethylsilyl)oxy)ethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)-4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzamide 10a 6g (30 mg, 0.040 mmol) was dissolved in dichloromethane (3 mL), and 7j (25 mg, 0.055 mmol) was added. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (17.5 mg, 0.091 mmol) and 4-dimethylaminopyridine (11.2 mg, 0.091 mmol) were then added in that order. After the addition was complete, the mixture was stirred at room temperature for 4 hours under a nitrogen atmosphere. 7j (16.6 mg, 0.037 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (10.5 mg, 0.055 mmol), and 4-dimethylaminopyridine (6.7 mg, 0.055 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction mixture was washed with water (5 mL), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using developing solvent system A to give the title product 10a (40 mg, yield: 79.9%). MS m / z (ESI): 1093.4 [M+1].

[0197] Step 2 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-4-((2-hydroxyethyl)(methyl)amino)-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 10 10a (40 mg, 0.036 mmol) was dissolved in tetrahydrofuran (2 mL), tetrabutylammonium fluoride (16 mg, 0.073 mmol) was added, and the mixture was stirred at room temperature for 4 h. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (8 mL) and washed with saturated ammonium chloride solution (10 mL × 2). The organic phase was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (8 mL) and washed with saturated ammonium chloride solution (10 mL × 6), water (10 mL), and saturated brine (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (HPLC) (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 10 (17 mg, yield: 47.5%). MS m / z (ESI): 979.2 [M+1]. 1H NMR (500 MHz, CD3OD) δ 8.26 (d, 1H), 8.04 (dd, 1H), 7.79 (d, 2H), 7.72 (d, 2H), 7.64 (d, 1H), 7.53 (d, 2H), 7.46 (t, 1H), 7.40-7.31 (m, 3H), 7.29-7.12 (m, 4H), 6.86-6.71 (m, 3H), 4.39 (d, 1H), 3.98 (br, 1H), 3.84-3.71 (m, 3H), 3.62 (d, 1H), 3.26-3.16 (m, 3H), 3.14-2.96 (m, 2H), 2.80-2.60 (m, 4H), 2.56-2.49 (m, 1H), 2.19 (t, 1H), 2.11-1.93 (m, 3H), 1.76 (d, 1H), 1.27-1.18 (m, 1H), 1.17-1.10 (m, 1H), 1.03-0.93 (m, 1H), 0.94-0.83 (m, 1H).

[0198] Example 2-11 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(3-hydroxypropyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 11 [ka] [ka] 8 (20 mg, 0.021 mmol) was dissolved in acetonitrile (2 mL), and triethylamine (22 mg, 0.22 mmol) and bromoethanol (30 mg, 0.22 mmol) were added, followed by the reaction mixture at 70 °C for 12 h under a nitrogen atmosphere. The mixture was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile), and the corresponding components were collected and concentrated under reduced pressure to give the title product 11 (13 mg, yield: 61.1%). MS m / z (ESI): 985.1 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.28 (d, 1H), 8.09 (dd, 1H), 7.86-7.78 (m, 2H), 7.63 (dd, 1H), 7.48-7.38 (m, 5H), 7.37-7.31 (m, 3H), 7.27 (dd, 2H), 7.23-7.17 (m, 2H), 6.81 (t, 3H), 4.44 (d, 1H), 4.08-4.00 (m, 1H), 3.81 (d, 1H), 3.72-3.55 (m, 4H), 3.25 (dd, 2H), 3.05 (s, 1H), 2.94 (s, 1H), 2.73-2.63 (m, 1H), 2.59-2.50 (m, 1H), 2.48-2.39 (m, 1H), 2.29 (s, 1H), 2.21 (t, 1H), 2.10-2.01 (m, 4H), 1.97-1.70 (m, 5H), 1.62 (s, 1H), 1.31-1.20 (m, 3H), 1.15 (d, 1H), 1.04-0.95 (m, 1H), 0.93 (d, 1H).

[0199] Example 2-12 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((S)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 12 [ka] [ka]

[0200] Step 1 tert-Butyl (S)-2-((R)-2-((4-(N-(4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylate 12a 8i (80 mg, 0.13 mmol) was dissolved in dichloromethane (3 mL), and 7j (70 mg, 0.15 mmol) was added. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (49 mg, 0.26 mmol) and 4-dimethylaminopyridine (32 mg, 0.26 mmol) were then added sequentially. After the addition was complete, the mixture was stirred at room temperature under a nitrogen atmosphere for 4 h. 7j (47 mg, 0.10 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (49 mg, 0.26 mmol), and 4-dimethylaminopyridine (32 mg, 0.26 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction mixture was washed with water (5 mL), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by thin-layer chromatography using developing solvent system A to give the title product 12a (80 mg, yield: 58.8%). MS m / z (ESI): 1059.0 [M-1].

[0201] Step 2 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((S)-pyrrolidin-2-yl)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 12 12a (50 mg, 0.047 mmol) was dissolved in dichloromethane (2 mL), 4 M hydrogen chloride in 1,4-dioxane (2 mL) was added, and the mixture was stirred at room temperature for 20 minutes. The reaction mixture was then concentrated under reduced pressure to remove the organic solvent. The residue was dissolved in dichloromethane and concentrated under reduced pressure three times. The resulting residue was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 12 (6 mg, yield: 66.2%). MS m / z (ESI): 961.2 [M+1]. 1H NMR (500 MHz, CD3OD) δ 8.23 ​​(s, 1H), 8.03 (d, 1H), 7.80 (d, 2H), 7.72 (d, 2H), 7.64 (d, 1H), 7.53 (d, 2H), 7.47 (t, 1H), 7.35 (t, 3H), 7.27-7.14 (m, 4H), 6.79 (d, 2H), 6.73 (d, 1H), 4.39 (d, 1H), 3.98 (s, 1H), 3.81-3.73 (m, 1H), 3.63-3.58 (m, 1H), 3.56-3.50 (m, 1H), 3.21-3.13 (m, 2H), 2.64 (t, 1H), 2.50 (t, 1H), 2.33-2.25 (m, 1H), 2.25-2.20 (m, 1H), 2.11-1.91 (m, 6H), 1.80-1.65 (m, 2H), 1.64-1.54 (m, 2H), 1.25-1.17 (m, 2H), 1.16-1.10 (m, 1H), 1.00-0.93 (m, 1H).

[0202] Example 2-13 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(2-hydroxyethyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 13 [ka] [ka] 12 (30 mg, 0.031 mmol) was dissolved in acetonitrile (4 mL), and triethylamine (32 mg, 0.31 mmol) and bromoethanol (39 mg, 0.31 mmol) were added, followed by the reaction mixture at 70°C for 12 hours under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (isolation conditions: column: Sharpsil-T Prep C18, mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding components were collected and concentrated under reduced pressure to give the title product 13 (25 mg, yield: 56.9%). MS m / z (ESI): 1005.1[M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.27 (d, 1H), 8.08 (dd, 1H), 7.85-7.79 (m, 2H), 7.74 (d, 2H), 7.68-7.63 (m, 1H), 7.55 (d, 2H), 7.50-7.46 (m, 1H), 7.42-7.35 (m, 3H), 7.29-7.18 (m, 4H), 6.87-6.69 (m, 3H), 4.41 (d, 1H), 4.03 (br, 1H), 3.83-3.73 (m, 3H), 3.67-3.56 (m, 2H), 3.25-3.18 (m, 2H), 3.14-2.86 (m, 2H), 2.72-2.59 (m, 1H), 2.58-2.47 (m, 1H), 2.46-2.36 (m, 1H), 2.33-2.23 (m, 1H), 2.10-1.97 (m, 4H), 1.96-1.87 (m, 1H), 1.84-1.72 (m, 2H), 1.66-1.55 (m, 1H), 1.28-1.19 (m, 1H), 1.16 (d, 1H), 1.03-0.95 (m, 1H), 0.95-0.89 (m, 1H).

[0203] Example 2-14 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((2R)-4-((2-hydroxyethyl)(methyl)amino)-3-methyl-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 14 [ka]

[0204] Example 2-15 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-5-((2-hydroxyethyl)(methyl)amino)-1-(phenylthio)pentan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 15 [ka]

[0205] Example 2-16 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-((R)-1-(2-hydroxyethyl)pyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 16 [ka]

[0206] Example 2-17 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((R)-1-methylpyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 17 [ka]

[0207] Example 2-18 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-methylpyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 18 [ka]

[0208] Example 2-19 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-methylpyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 19 [ka]

[0209] Example 2-20 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(2-hydroxyethyl)-2-methylpyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 20 [ka]

[0210] Example 2-21 4-(4-((R)-hydroxy(4'-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(2-hydroxyethyl)-2-methylpyrrolidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 21 [ka]

[0211] Example 2-22 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((2R)-1-(1-(2-(hydroxyethyl)azetidin-2-yl)-3-(phenylthio)propan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide 22 [ka]

[0212] Example 2-23 [ka]

[0213] Example 2-24 AZD4320 [ka] It was prepared with reference to Example 3 (positive control compound AZD4320) on pages 160-161 of Patent WO2012017251.

[0214] Example 3-1 LD-1 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino)-3-methylbutyrylamino)-5-ureidopentanoylamino)benzyl (R)-2-((R)-2-((4-(N-(4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylic acid ester LD-1 [ka] [ka] 1 (3 mg, 0.003 mmol) was dissolved in 0.5 mL of N,N-dimethylformamide, and 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino)-3-methylbutyrylamino)-5-ureidopentanoylamino)benzyl (4-nitrophenyl) carbonate LD-1a (3.1 mg, 0.004 mmol, Hyo-yuan) was added. 0.1 mL of pyridine was added, and the mixture was purged with argon gas three times. 1-Hydroxybenzotriazole (2 mg, 0.014 mmol) and N,N-diisopropylethylamine (2 mg, 0.015 mmol) were added, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was purified by high-performance liquid chromatography (isolation conditions: column: XBridge Prep C18, mobile phase: ammonium acetate, water, acetonitrile), and the corresponding components were collected and concentrated under reduced pressure to give the title product LD-1 (2 mg, yield: 40.5%). MS m / z (ESI): 1525.0 [M+1]. 1 H NMR (500 MHz, CD3OD) δ 8.17 (s, 1H), 7.92-7.80 (m, 3H), 7.64-7.51 (m, 3H), 7.46-7.09 (m, 12H), 6.84-6.74 (m, 3H), 6.48-6.40 (m, 2H), 5.35 (t, 4H), 4.41 (d, 1H), 4.15 (d, 1H), 4.00-3.71 (m, 6H), 3.66-3.54 (m, 2H), 3.44 (t, 2H), 3.22-3.05 (m, 6H), 2.77 (t, 1H), 2.72-2.62 (m, 2H), 2.53 (t, 2H), 2.23 (t, 2H), 2.18 (t, 4H), 2.11-2.06 (m, 2H), 1.96-1.82 (m, 4H), 1.80-1.71 (m, 2H), 1.65-1.51 (m, 8H), 1.17-1.09 (m, 2H), 1.00-0.89 (m, 9H).

[0215] Biological evaluation Test Example 1: Measurement of the binding activity of compounds according to the present disclosure to BCL-xL In vitro BCL-xL binding activity was tested by the following method.

[0216] Reagents used in this experiment: His-tagged BCL-xL protein (R&D, product number 894-BX-050), biotin-labeled BIM protein (R&D, product number 3526 / 1), binding buffer (cisbio, product number 62DLBDDF), detection buffer (cisbio, product number 62DB1FDG), europium cryptate-labeled anti-6His antibody (cisbio, product number 61HI2KLA), and streptavidin XL665 (cisbio, product number 611SAXLA).

[0217] The following in vitro binding assay can measure the binding activity of a test compound to BCL-xL.

[0218] Test compounds were dissolved in dimethyl sulfoxide and diluted to various concentrations as needed for the experiment. Each concentration of compound prepared in dimethyl sulfoxide was further diluted 100-fold in binding buffer. BCL-xL was diluted to the corresponding concentration in binding buffer and added to a white 384-well plate at 4 μL / well. Further diluted test compounds were added to the white 384-well plate at 2 μL / well, mixed uniformly, and incubated at 25°C for 1 hour. Biotin-labeled BIM protein was diluted to the corresponding concentration in binding buffer and added to a white 384-well plate at 4 μL / well, mixed uniformly, and incubated at 25°C for 2 hours. Europium cryptate-labeled anti-6His antibody and streptavidin XL665 were diluted to the corresponding concentration in detection buffer and added to a white 384-well plate at 5 μL / well, mixed uniformly, and incubated at 25°C for 2-4 hours. The signal values ​​were read using an HTRF program on a microplate reader (BMG Labtech, model number PHERAstar FS).

[0219] The test results are shown in Table 1. [Table 2] Conclusion: The compounds of the present disclosure have high binding activity to BCL-xL.

[0220] Test Example 2: Measurement of the binding activity of compounds according to the present disclosure to BCL-2 In vitro BCL-2 binding activity was tested by the following method.

[0221] Reagents used in this experiment: His-tagged BCL-2 protein (R&D Systems, product number 827-BC-050), biotin-labeled BIM protein (R&D, product number 3526 / 1), binding buffer (cisbio, product number 62DLBDDF), detection buffer (cisbio, product number 62DB1FDG), europium cryptate-labeled anti-6His antibody (cisbio, product number 61HI2KLA), and streptavidin XL665 (cisbio, product number 611SAXLA).

[0222] The in vitro binding assay described below can measure the binding activity of a test compound to BCL-2.

[0223] Test compounds were dissolved in dimethyl sulfoxide and diluted to various concentrations as needed for the experiment. Each concentration of compound prepared in dimethyl sulfoxide was further diluted 100-fold in binding buffer. BCL-2 was diluted to the corresponding concentration in binding buffer and added at 4 μL / well to a white 384-well plate. The diluted test compound was then added at 2 μL / well to the white 384-well plate, mixed uniformly, and incubated at 25°C for 1 hour. Biotin-labeled BIM protein was diluted to the corresponding concentration in binding buffer and added at 4 μL / well to the white 384-well plate, mixed uniformly, and incubated at 25°C for 2 hours. Europium cryptate-labeled anti-6His antibody and streptavidin XL665 were diluted to the corresponding concentration in detection buffer and added at 5 μL / well to the white 384-well plate, mixed uniformly, and incubated at 25°C for 2-4 hours. The signal values ​​were read using an HTRF program on a microplate reader (BMG Labtech, model number PHERAstar FS).

[0224] The test results are shown in Table 2. [Table 3] Conclusion: The compounds of the present disclosure have high binding activity to BCL-2.

[0225] Test Example 3: Measurement of the growth inhibitory activity of compounds according to the present disclosure on RS4;11 cells and NCI-H146 cells The in vitro inhibitory activity against RS4;11 cells and NCI-H146 cells was tested by the following method.

[0226] Reagents used in this experiment: RPMI-1640 medium (HyCLone, product number SH30243018), fetal bovine serum (Gibco, product number 10099-141), pancreatin (Gibco, product number 25200-072), and Cell Titer-Glo (Promega, product number G7571).

[0227] Materials used: RS4;11 cells (Nanjing Kebei Biotechnology Co., Ltd., product number CBP60641), NCI-H146 cells (ATCC, product number HTB-173).

[0228] The following in vitro cell proliferation experiment can measure the growth inhibitory activity of test compounds on RS4;11 cells and NCI-H146 cells.

[0229] RS4;11 cells (or NCI-H146 cells) were cultured in RPMI-1640 medium containing 10% FBS and passaged two to three times a week at a passage ratio of 1:3 or 1:5. For passage, the cells were transferred to a centrifuge tube and centrifuged at 1200 rpm for 3 minutes. The remaining supernatant medium was discarded, and fresh medium was added to resuspend the cells. 90 μL of the cell suspension was placed in a 96-well cell culture plate, and the density of RS4;11 cells and NCI-H146 cells was 2.22 × 10 5 The cells / mL were added to the periphery of the 96-well plate with 100 μL of complete medium (RPMI-1640 medium containing 10% FBS). The culture plate was cultured in an incubator (37°C, 5% CO2) for 24 hours.

[0230] Depending on the experimental needs, samples to be assayed were diluted to an appropriate concentration gradient with dimethyl sulfoxide. 5 μL of the compound solution to be assayed was added to 95 μL of fresh medium and mixed uniformly. Then, 10 μL of the compound-containing medium solution was added to a cell culture plate. The culture plate was incubated in an incubator (37°C, 5% CO2) for 3 days. 50 μL of CellTiter-Glo reagent was added to each well of a 96-well cell culture plate and left at room temperature in the dark for 5–10 minutes. Chemiluminescence signal values ​​were read using a microplate reader (BMG Labtech, model PHERAstar FS), and the data were processed using GraphPad software. The results are shown in Table 3. [Table 4] Conclusion: The compounds of the present disclosure have high growth inhibitory activity against RS4;11 and NCI-H146 cells.

[0231] Test Example 4: Toxicity test of repeated administration to SD rat intravenous for 7 days 1. Purpose of the experiment The toxicity of Compound 9 and AZD4320 was evaluated in SD rats by repeated intravenous administration for 7 days.

[0232] 2. Experimental drugs Storage conditions: Stored in a dark place in a desiccator at room temperature. Compound 9 has a molecular weight of 976.1 when assumed to be 100% pure. The compound AZD4320 has a molecular weight of 945.5 when assumed to be 100% pure.

[0233] 3. Experimental design and methods 3.1 Experimental animals and breeding conditions Thirty SD rats, weighing approximately 180–200 g and aged 6–7 weeks, were purchased, half male and half female, from Zhejiang Weitong Lihua Experimental Animal Co., Ltd. and were SPF-grade. Animal certification numbers were 200917Aazz0619000298 and 20200917Aazz0619000104. Production license number was SCXK(Zhejiang)2019-0001. Housing conditions: Animals were housed in SPF-grade transparent plastic rat cages, with 2-3 animals per cage. The room temperature was 20-26°C, and the relative humidity was 40-70%. The conditions were controlled with a 12-hour light / 12-hour dark alternating cycle. Mouse pellet chow was available ad libitum.

[0234] 3.2. Animal Grouping The animals were randomly divided into three groups: a solvent control group, compound 9-1 mg / kg, and compound AZD4320-1 mg / kg, with six animals per group, half of which were female and half were male.

[0235] 3.3 Experimental Method The dose of the test substance in this 7-day toxicity test on rats was 1 mg / kg (qd), administered for 7 consecutive days. The administration route was intravenous, the administration volume was 10 mL / kg, and the administration frequency and duration were daily morning administration for 7 consecutive days. Solvent: 1.5% DMA + 98.5% (10% HS-15), where DMA is N,N-dimethylacetamide, Shanghai Taitan Technology Co., Ltd., product number 01013630, and HS-15 is polyoxyethylene 15 stearate (Solutol HS15), Shanghai Xietai Chemical Co., Ltd., product number 50253404.

[0236] 3.4. Experimental equipment Hitachi 7100 fully automatic biochemical analyzer Siemens advia 2120i fully automated hematology analyzer Sysmex ca1500 fully automatic blood coagulation analyzer Detection of toxicokinetic data: AB 4000 liquid chromatograph mass spectrometer.

[0237] 3.5 Data representation and statistical processing The test results are entered into Excel and statistically processed. Clinical symptoms, food intake and dissection were roughly observed visually without any statistical calculations, and indices such as body weight, organ weight and clinical examination were statistically analyzed using the T-test.

[0238] 4, results SD rats were intravenously administered 1 mg / kg of compound 9 and AZD4320 at 5 mL / kg repeatedly for 7 days: During the administration period, no clinically significant abnormalities were observed in the animals in the compound 9 administration group. Compared with the vehicle control group, no significant differences were observed in body weight and body weight gain rate (p>0.05) in the animals in each dose group, and no significant changes were observed in food intake.

[0239] Rats treated with Compound 9 had a slight reduction in platelets, but these were within the normal range, whereas all AZD4320 rats showed a significant reduction in platelets (p<0.01). The results are shown in Table 4. [Table 5]

[0240] 5. Conclusion Under the experimental conditions, Compound 9 has the advantage of being safer and causing less platelet reduction than the solvent control group and the positive control AZD4320 group.

[0241] Test Example 5: Pharmacokinetic evaluation of compounds according to the present disclosure in mice 1. Summary Mice were used as test animals, and compound 10 and AZD4320 were administered intravenously (iv) to the mice, and the plasma drug concentrations were measured at different time points using LC / MS / MS. The pharmacokinetic behavior of the compound according to the present disclosure in mice was studied, and its pharmacokinetic characteristics were evaluated.

[0242] 2. Test plan 2.1 Study Drug compound 10

[0243] 2.2 Test animals Eighteen female C57 mice were randomly divided into two groups and purchased from Zhejiang Weitong Lihua Laboratory Animal Co., Ltd., with the animal production license number: SCXK(Hu)2017-0005.

[0244] 2.3 Drug Preparation A certain amount of compound 10 was weighed, dissolved in 5% DMSO and 5% Tween 80 (Shanghai Taitan Technology Co., Ltd.), and then added with 90% saline to prepare a 0.1 mg / mL clear solution. Compound AZD4320 was prepared in the same manner as compound 10.

[0245] 2.4 Administration Intravenous administration: Compound 10 and AZD4320 were each intravenously administered to mice, with the dose of each being 2.0 mg / kg and the administration volume being 20.0 mL / kg.

[0246] 3, operation Compound 10 was administered intravenously to mice, and 0.1 mL of blood was collected before and 5 minutes after administration, and at 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 11.0, and 24.0 hours. The collected blood was placed in an EDTA-K2 anticoagulant tube and centrifuged at 10,000 rpm for 1 minute (4°C). Plasma was separated within 1 hour and stored at -20°C for measurement. The entire process, from blood collection to centrifugation, was carried out under ice bath conditions.

[0247] Mice were intravenously administered the compound AZD4320, and 0.1 mL of blood was collected before and 5 minutes after administration, and at 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 11.0, and 24.0 hours. The collected blood was placed in an EDTA-K2 anticoagulant tube and centrifuged at 10,000 rpm for 1 minute (4°C). Plasma was separated within 1 hour and stored at -20°C for measurement. The entire process, from blood collection to centrifugation, was carried out under ice bath conditions.

[0248] The amount of compound to be measured in the plasma of mice administered different concentrations of drugs was measured as follows: 10 μL of mouse plasma was taken at each post-administration time point, and 200 μL of methanol (containing the internal standard solution of camptothecin (100 ng / mL)) was added. The mixture was vortex-mixed for 1 minute and centrifuged for 10 minutes (18,000 rpm). 3 μL of the supernatant was then taken from the plasma sample and analyzed by LC / MS / MS.

[0249] 4. Pharmacokinetic parameter results [Table 6] Conclusion: Compound 10 of the present disclosure exhibits slow clearance in mice and a longer clearance phase half-life than the positive control AZD4320, demonstrating pharmacokinetic advantages.

[0250] Test Example 6: Therapeutic effect on human acute lymphoblastic leukemia RS4;11 tumors subcutaneously transplanted into mice This experiment was conducted to evaluate and compare the therapeutic effects of compounds according to the present disclosure on subcutaneously transplanted human acute lymphoblastic leukemia RS4;11 tumors in mice.

[0251] 1. Experimental animals and breeding conditions Five-week-old female NOD-Scid mice were purchased from Beijing Huafukang Biotechnology Co., Ltd. Production license number: SCXK(Kyoto)2019-0008, Animal Qualification Certificate number: 110322211100992176. The breeding environment was SPF grade. There were six mice in the vehicle group and eight mice in the compound group.

[0252] II. Test Compound Compound 9, compound 10 and solvent group. Five mg of compound was placed in 250 μL of DMSO and dissolved by vortexing to obtain a 20 mg / mL solution, which was then diluted to the desired concentration with 30% hydroxypropyl-β-cyclodextrin (HP-β-CD, Yinusi Biotechnology Nantong Co., Ltd.) solution.

[0253] III. Experimental Design and Methods Human acute lymphoblastic leukemia RS4:11 was purchased from the American Type Culture Collection. RS4:11 cells were cultured in 10-cm culture dishes in RPMI 1640 medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), penicillin, and streptomycin at 37°C in a 5% CO2 incubator. Cells were passaged twice weekly, and when in exponential growth phase, they were harvested, counted, and inoculated. Inject 1 x 10 subcutaneously into each mouse. 7 RS4;11 cells were inoculated and tumors were grown to 100–200 mm 3 Once the tumors reached the tumor size, they were divided into groups based on tumor volume and injected with the drug intravenously (IV) once a week (QW) for a total of four times, with an injection volume of 0.1 mL / 10 g body weight and a dose of 10 mg / kg.

[0254] 4. Results The experimental endpoints are used to examine the effects of drugs on tumor growth, and the specific endpoints are T / C% or tumor growth inhibition rate (TGI%). Tumor diameters were measured twice a week with a vernier caliper, and tumor volume (V) was calculated as follows: V = 1 / 2 × a × b 2 Among them, a and b represent length and width respectively. T / C(%) = (T - T0) / (C - C0) × 100. Among them, T and C are the tumor volumes at the end of the experiment, and T0 and C0 are the tumor volumes at the start of the experiment. Tumor growth inhibition rate (TGI)(%) = 100 - T / C(%). When tumor regression appears, tumor growth inhibition rate (TGI)(%) = 100 - (T - T0) / T0 × 100. When the tumor becomes smaller than the initial volume, that is, when T < T0 or C < C0, it is defined as partial response (PR), and when the tumor completely disappears, it is defined as complete response (CR).

Table 7

Claims

1. A compound represented by general formula (D) or a pharmaceutically acceptable salt thereof, 【Chemistry 1】 Among them, R １ is a hydrogen atom, R ２ is a C 1-6 alkyl group, Or, R １ and R ２ together with the atoms to which they are attached form a 3- to 6-membered heterocyclyl group; R ３ is selected from a hydrogen atom, a C 1-6 hydroxyalkyl group, and a C 1-6 alkyl group; R ４ is selected from a hydrogen atom and a C 1-6 alkyl group, R ５ is selected from halogen and C 1-6 haloalkyl groups; R ６ is selected from a hydroxy group and a C 1-6 alkoxy group; R ７ is a hydrogen atom, R ８ and R ９ are each independently selected from a hydrogen atom and a C 1-6 alkyl group; R １０ and R １１ are each independently selected from a hydrogen atom and a C 1-6 alkyl group; R １２ is a hydrogen atom, R １３ is a hydrogen atom, R １４ is a hydrogen atom, r is selected from 0, 1, 2 and 3; As a condition, R １ and R ２ When R does not form a 3- to 6-membered heterocyclyl group together with the atom to which it is attached, ５ is a C 1-6 haloalkyl group; A compound represented by the general formula (D) or a pharmaceutically acceptable salt thereof:

2. The R １ and R ２ and R 1 and R 2 together with the atom(s) linked thereto form a pyrrolidinyl group, or a pharmaceutically acceptable salt thereof.

3. A compound represented by general formula (DI) or a pharmaceutically acceptable salt thereof, 【Chemistry 2】 wherein m is selected from 0, 1, 2 and 3; r, and R ３ ~R １４ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

4. A compound represented by general formula (D-II) or a pharmaceutically acceptable salt thereof, 【Transformation 3】 wherein m is selected from 0, 1, 2 and 3; r, and R ３ ~R １１ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

5. A compound represented by general formula (D-III) or a pharmaceutically acceptable salt thereof, 【Chemistry 4】 wherein m is selected from 0, 1, 2 and 3; r, and R ３ ~R １１ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

6. A compound represented by general formula (D-IV) or a pharmaceutically acceptable salt thereof, 【Transformation 5】 Among them, m is selected from 0, 1, 2 and 3; R ３ ~R ９ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

7. 4. The compound represented by formula (D) according to claim 3, or a pharmaceutically acceptable salt thereof, wherein m is 2.

8. The R ５ The compound represented by formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is a chlorine atom.

9. The R ５ The compound represented by formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is a trifluoromethyl group.

10. The R ３ is a C 1-6 hydroxyalkyl group, or a pharmaceutically acceptable salt thereof.

11. A compound represented by general formula (D-V) or a pharmaceutically acceptable salt thereof, 【Transformation 6】 wherein t is selected from 0, 1, 2, and 3; r, R １ , R ２ , R ４ , and R ６ ~R １４ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

12. A compound represented by general formula (D-VI) or a pharmaceutically acceptable salt thereof, 【Transformation 7】 wherein t is selected from 0, 1, 2, and 3; r, R １ , R ２ , R ４ , and R ６ ~R １１ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

13. A compound represented by general formula (D) according to claim 12 or a pharmaceutically acceptable salt thereof, which is represented by general formula (D-VI) or a pharmaceutically acceptable salt thereof, wherein t is 1.

14. A compound represented by general formula (D-VII) or a pharmaceutically acceptable salt thereof, 【Transformation 8】 Among them, r and R １ ~R １１ is as defined in claim 1 A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

15. The R ４ The compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein is a hydrogen atom.

16. The R ６ The compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: is a hydroxy group.

17. The R ７ , R ８ , R ９ 2. The compound represented by formula (D) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R 10 and R 11 are hydrogen atoms.

18. The compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein r is 0 or 1.

19. The compound represented by general formula (D) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein r is 0. 【Request Item 20】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 or a pharmaceutically acceptable salt thereof, A compound represented by the general formula (D) according to claim 1 or a pharmaceutically acceptable salt thereof.

21. A method for preparing a compound represented by general formula (D-III) or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 12】 a compound of general formula (DA2) or a pharmaceutically acceptable salt thereof is subjected to a substitution reaction under basic conditions to obtain a compound of general formula (D-III) or a pharmaceutically acceptable salt thereof, wherein the reagent used under the basic conditions is triethylamine; Among them, m is selected from 0, 1, 2 and 3; R ３ is a C 1-6 hydroxyalkyl group, r, and R ４ ~R １１ is as defined in claim 1 method.

22. A pharmaceutical composition comprising a therapeutically effective amount of a compound represented by general formula (D) according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable vector, diluent or excipient.

23. 23. The pharmaceutical composition of claim 22 for use in inhibiting Bcl-2 and / or Bcl-xL.

24. The pharmaceutical composition of claim 22 for use in treating cancer or tumors, wherein the cancer is selected from the group consisting of melanoma, liver cancer, renal cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, nasopharyngeal cancer, colorectal cancer, colon cancer, rectal cancer, pancreatic cancer, cervical cancer, ovarian cancer, breast cancer, bladder cancer, prostate cancer, leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, acute lymphocytic leukemia, acute granulocytic leukemia, mantle cell lymphoma, diffuse large B-cell lymphoma, follicle center lymphoma, non-Hodgkin's lymphoma, T-cell lymphoma, B-cell lymphoma, squamous cell carcinoma of the head and neck, cervical cancer, thyroid cancer, lymphoma, sarcoma, neuroblastoma, brain tumor, myeloma, astrocytoma, and glioma.

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