Use of eribulin derivative drug conjugate in treatment of tumor

Abstract

The present invention provides a use of an Eribulin derivative drug conjugate in the treatment of a tumor. Particularly, the present invention provides a use of a compound as represented by formula IA or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a tumor disease resistant to an anti-HER2 antibody-drug conjugate.

Description

Use of eribulin derivative drug conjugates for treating tumors Technical Field

[0001] The present disclosure relates to the field of biomedicine, and in particular to the use of eribulin derivative drug conjugates for preparing tumor treatment. Background Art

[0002] Antibody drug conjugates (ADCs) link monoclonal antibodies or antibody fragments to biologically active drugs via stable chemical linker compounds. They leverage the specificity of antibodies for binding to surface antigens on both normal and tumor cells and the high efficacy of drugs, while avoiding the drawbacks of the former, such as low efficacy, and the latter's excessive toxic side effects. This means that compared to traditional chemotherapy drugs, ADCs can precisely bind to tumor cells while minimizing their effects on normal cells.

[0003] Microtubules are powerful, filamentous cytoskeletal proteins associated with a variety of cellular functions, including intracellular migration and transport, cell signaling, and maintaining cell shape. Microtubules also play a key role in mitotic cell division by forming the mitotic spindle, which is required for the division of chromosomes into two daughter cells. The biological functions of microtubules in all cells are largely regulated by their polymerization dynamics, which is achieved by the reversible, non-covalent addition of α and β tubulin dimers to the ends of the microtubules. This dynamic behavior and the resulting control of microtubule length are indispensable for the proper function of the mitotic spindle. Even slight changes in microtubule dynamics can involve the axis checkpoint, inhibiting cell cycle progression during mitosis and subsequently causing cell death. Because cancer cells divide rapidly, they are generally more sensitive to compounds that bind to tubulin and disrupt its normal functions compared to normal cells. Therefore, tubulin inhibitors and their antibody-drug conjugates are expected to become a promising class of drugs for treating cancer.

[0004] Currently, clinically available ADCs targeting HER2 (such as DS-8201) have shown promising clinical efficacy in patients with HER2-expressing or mutated cancers. However, most of them ultimately lead to disease progression due to different mechanisms. Summary of the Invention

[0005] The present disclosure provides a use of a compound represented by Formula IA or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating tumor diseases resistant to anti-HER2 antibody-drug conjugates.

[0006] in:

[0007] Ab is Pertuzumab;

[0008] n is 1 to 10.

[0009] In some embodiments, the range of drug loading (n) can be the average number of cytotoxic drugs bound to each pertuzumab antibody, non-limiting examples include the average number of cytotoxic drugs bound to each antibody is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and any range between these points. For example, it can be 2-8, 2-7, 2-6, 2-5, 2-4, 3-4, 3-5, 3.5 to 4.7, 5-6, 5-7, 5-8 and 6-8. Exemplary, the drug loading (n) can be the average of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. n is a decimal or an integer. In some embodiments, n is 1 to 8, or 3 to 5.

[0010] The anti-HER2 antibody-drug conjugate-resistant tumor described in the present disclosure refers to a tumor disease that becomes resistant after treatment with an anti-HER2 antibody-drug conjugate. The tumor disease may be breast cancer, gastric cancer, colorectal cancer, urothelial carcinoma, bile duct cancer, lung cancer (e.g., non-small cell lung cancer), etc.

[0011] In some embodiments, the tumor disease can be a HER2-positive, HER2-underexpressing, HER2-overexpressing, or HER2-mutant tumor disease.

[0012] In some embodiments, the anti-HER2 antibody drug conjugate is selected from the compound represented by Formula II or a pharmaceutically acceptable salt thereof, Ab-(LD) k (II)

[0013] in,

[0014] -D represents the residue of exitecan or its derivatives;

[0015] L is a linker;

[0016] Ab is anti-HER2 antibody;

[0017] k is 1 to 10. In some embodiments, k is 1 to 8, or 3 to 5.

[0018] In some embodiments, -D is:

[0019] in,

[0020] A is -C(O)NH- or -O-;

[0021] Every R 1a 、R 1b Each independently selected from a hydrogen atom, C 1-6alkyl, 3-6 membered cycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the alkyl, cycloalkyl, aryl or heteroaryl is optionally substituted with one or more substituents selected from C1-C6 alkyl, halogen, hydroxy, amino, oxo, 3-6 membered cycloalkyl, 6-10 membered aryl or C1-C6 alkoxy, or R 1a and R 1b Together with the carbon atom to which it is attached, it forms a 3-6 membered cycloalkyl group which is optionally substituted by one or more substituents selected from C1-C6 alkyl, halogen, hydroxy, amino, oxo or C1-C6 alkoxy;

[0022] R 1c Selected from hydrogen atoms, deuterium atoms, C 1-6 Alkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the alkyl, aryl and heteroaryl are optionally substituted with one or more substituents selected from C1-C6 alkyl, halogen, hydroxyl, amino and oxo;

[0023] n1 is an integer from 0 to 10, for example, an integer from 0 to 6.

[0024] In some embodiments, -D is:

[0025] In some embodiments, the linker is stable outside the cell, such that the ADC remains intact when present in the extracellular environment, but is capable of cleavage upon internalization in a cell, such as a cancer cell. In some embodiments, when the ADC enters a cell expressing an antigen specific for the antibody portion of the ADC, the drug portion is cleaved from the antibody portion, and the cleavage releases the unmodified form of the drug.

[0026] In some embodiments, the cleavable portion in the linker is a cleavable peptide portion. In some embodiments, relative to ADCs comprising other cleavable portions, ADCs comprising cleavable peptide portions show lower aggregation levels, improved antibody to drug ratios, increased targeted killing of cancer cells, reduced off-target killing of non-cancerous cells, and / or higher drug load (p). In some embodiments, relative to non-cleavable linkers, adding a cleavable portion increases cytotoxicity and / or efficacy. In some embodiments, the increased efficacy and / or cytotoxicity is increased efficacy and / or cytotoxicity in cancers that express moderate levels of the antigen targeted by the antibody portion of the ADC (e.g., moderate FRA expression). In some embodiments, the cleavable peptide portion is capable of being cleaved by an enzyme, and the linker is a linker that the enzyme is capable of cleaving. In some embodiments, the enzyme is a cathepsin, and the linker is a linker that the enzyme is capable of cleaving. In certain embodiments, compared to other cleavage mechanisms, linkers that the enzyme is capable of cleaving (e.g., linkers that the cathepsin is capable of cleaving) show one or more of the above-mentioned improved properties.

[0027] In some embodiments, the linker comprises an amino acid unit, which preferably comprises a peptide residue consisting of 2 to 7 amino acids selected from phenylalanine, glycine, valine, lysine, citrulline, serine, glutamic acid, and aspartic acid, more preferably valine-citrulline (Val-Cit), alanine-alanine-asparagine (Ala-Ala-Asn), glycine-glycine-lysine (Gly-Gly-lys), valine-lysine (Val-lys), valine-alanine (Val-Ala), valine-phenylalanine (Val-Phe) or glycine-glycine-phenylalanine-glycine (Gly-Gly-Phe-Gly).

[0028] In some embodiments, the present disclosure comprises a linker of an amino acid unit selected from the group consisting of:

[0029] In some embodiments, the amino acid unit comprises valine-citrulline (Val-Cit). In some embodiments, ADCs comprising Val-Cit exhibit increased stability, reduced off-target cell killing, increased on-target cell killing, lower aggregation levels, and / or higher drug loading relative to ADCs comprising other amino acid units or other cleavable moieties.

[0030] In another aspect, some embodiments provide a linker comprising a cleavable sulfonamide moiety, wherein the linker is cleavable under reducing conditions.

[0031] In some embodiments, the linker comprises a cleavable disulfide moiety, and the linker is capable of being cleaved under reducing conditions.

[0032] In another aspect, the linker in the disclosed antibody conjugates comprises at least one spacer unit that connects the drug D to the cleavable moiety. In some embodiments, the linker comprises a spacer unit connected to D.

[0033] In some embodiments, the spacer unit comprises p-aminobenzyloxycarbonyl (PAB),

[0034] In some embodiments, the Spacer unit comprises a p-aminobenzoyl group,

[0035] In some embodiments, the spacer unit comprises:

[0036] Wherein, Z1 to Z5 are each independently selected from a carbon atom or a nitrogen atom; R 14R is selected from alkyl, cycloalkyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally substituted by one or more substituents selected from alkyl, alkoxy, halogen, deuterium, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl; 11 and R 12 Each independently selected from hydrogen, deuterium, C 1-6 Alkyl, C 3-6 Cycloalkyl, preferably hydrogen; or, R 11 With R 12 Together with the carbon atom to which it is attached, it forms a C3-6 cycloalkyl group; X is selected from -O- or -NH-; L is selected from an integer between 1 and 4;

[0037] Q is VE-, which provides a glycosidic bond that can be cleaved by intracellular glycosidases, and E is selected from -O-, -S-, or -NR 13 -, R 13 is selected from hydrogen or methyl, further, V is selected from where R 15 In some embodiments, V is selected from -COOH or CH2OH.

[0038] In some embodiments, the spacer unit comprises:

[0039] Z1, Z3, Z4, X, Q, R 11 、R 12 、R 14 As mentioned above.

[0040] In some embodiments, the Spacer unit comprises a moiety selected from:

[0041] -(CR a R b ) m1 -O(CR a R b ) m2 -CR 8 R 9 -C(O)-,

[0042] -(CR a R b ) m1 NH-(CR a R b ) m2 -CR 8 R 9 -C(O)-,

[0043] -(CR a R b )m1 O-CR 8 R 9 (CR a R b ) m2 -,

[0044] -(CR a R b ) m1 OCR 8 R 9 -C(O)-,

[0045] -(CR a R b ) m1 -O-(CR a R b ) m2 C(O)-or-(CR a R b ) m1 -S-(CR a R b ) m2 -CR 8 R 9 -C(O)-,

[0046] where R a and R b are the same or different and are each independently selected from hydrogen, a deuterium atom, a halogen or an alkyl group; R 8 Selected from hydrogen, C 3-6 Cycloalkylalkyl or C 3-6 Cycloalkyl; R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen; or, R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl; m1 and m2 are each independently selected from 0, 1, 2 or 3.

[0047] In some embodiments, the Spacer unit comprises a moiety selected from:

[0048] -(CH2)3-C(O)-, -CH2-O-CH2-C(O)-, -(CH2)2-O-CH2-C(O)-,

[0049] In another aspect, in the disclosed antibody conjugate (ADC), LD is a chemical moiety represented by the following formula:

[0050] -Str-(Pep)-Sp-D

[0051] Str is a stretching unit covalently linked to Ab.

[0052] Sp is a spacer unit,

[0053] Pep is selected from an amino acid unit, a disulfide moiety, a sulfonamide moiety, or the following non-peptide chemical moieties:

[0054] Wherein W is -NH-heterocycloalkyl- or heterocycloalkyl; Y is heteroaryl, aryl, -C(O)C 1-6 Alkylene, C 2-6 Alkenylene, C 1-6 Alkylene or -C 1-6 Alkylene-NH-;

[0055] Each R 2 Independently selected from C 1-10 Alkyl, C 2-10 Alkenyl, C 1-6 Alkylene-NH2, -(C 1-10 Alkylene) NHC(NH)NH2 or -(C 1-10 Alkylene)NHC(O)NH2;

[0056] R 3 and R 4 Each is independently H, C 1-10 Alkyl, C 2-10 alkenyl, arylalkyl, heteroarylalkyl, or R 3 and R 4 Together they can form C 3-7 Cycloalkyl;

[0057] R 5 and R 6 Each is independent of C 1-10 Alkyl, C 2-10 Alkenyl, arylalkyl, heteroarylalkyl, (C 1-10 Alkylene) OCH2-, or R 5 and R 6 Together they can form C 3-7 Cycloalkyl ring.

[0058] In some embodiments, Y in the antibody-drug conjugate (ADC) is selected from the following moieties:

[0059] In another aspect, in the antibody-drug conjugate (ADC), Str is selected from a chemical moiety represented by the following formula:

[0060] where R 7 Selected from -W1-C(O)-, -C(O)-W1-C(O)-, -(CH2CH2O)p1 C(O)-、-(CH2CH2O) p1 CH2C(O)-、-(CH2CH2O) p1 CH2CH2C(O)-, wherein W1 is selected from C 1-8 Alkylene, C 1-8 Alkylene-cycloalkyl or straight chain heteroalkyl of 1 to 8 atoms, said heteroalkyl containing 1 to 3 heteroatoms selected from N, O or S, wherein said C 1-8 Alkylene, cycloalkyl and straight-chain heteroalkyl are each independently optionally further substituted with one or more substituents selected from halogen, deuterium, hydroxy, cyano, amino, alkyl, haloalkyl, deuterated alkyl, alkoxy and cycloalkyl;

[0061] L 1 Selected from-NR 10 (CH2CH2O) p1 CH2CH2C(O)-、-NR 10 (CH2CH2O) p1 CH2C(O)-、-S(CH2) p1 C(O)-、-(CH2) p1 C(O)- or chemical bond, preferably chemical bond; wherein p1 is an integer from 1 to 20, R 10 is selected from the group consisting of a hydrogen atom, an alkyl group, a halogenated alkyl group, a deuterated alkyl group, and a hydroxyalkyl group.

[0062] In some embodiments, C 1-8 Alkylene-cycloalkyl is selected from methylene-cyclohexyl Ethylene-cyclohexyl Methylene-cyclopentyl

[0063] In some embodiments, the linker can comprise at least one polyethylene glycol (PEG) moiety. The PEG moiety can, for example, comprise -(PEG) p1 -, Where p1 is an integer from 1 to 20, for example (PEG)2; (PEG)4; (PEG)5.

[0064] In some embodiments, the stretcher in the linker comprises (PEG) 2. In some embodiments, despite the shorter linker length, ADCs comprising shorter stretchers (e.g., (PEG) 2) exhibit lower aggregation levels and / or higher drug loading relative to ADCs comprising longer stretchers (e.g., (PEG) 8).

[0065] In some embodiments, Str of the antibody-drug conjugate is Medium R 7 Selected from C 1-6 Alkylene C(O)-, -(CH2-CH2O)2C(O)-, -(CH2-CH2O)2CH2C(O)-, -(CH2-CH2O)2CH2CH2C(O)-, -(CH2-CH2O)2CH2CH2C(O)-, -(CH2-CH2O)3C(O)- and -(CH2-CH2O)4C(O)-.

[0066] In some embodiments, Str of the antibody-drug conjugate is Medium R 7 Selected from -C 1-8 Alkylene-cycloalkyl-C(O)-, -(CH2-CH2O)4CH2C(O)- and -(CH2-CH2O)6CH2C(O)-.

[0067] In some embodiments, the linker L in the antibody-drug conjugate comprises: maleimide-(PEG)2-Val-Cit, maleimide-(PEG)6-Val-Cit, maleimide-(PEG)8-Val-Cit, maleimide-(PEG)4-CH2CH2C(O)-Val-lys, maleimide-(CH2)5-Val-Cit, maleimide-(CH2 )5-Val-lys, Maleimide-(CH2)5-Gly-Gly-Phe-Gly, Maleimide-(PEG)2-Ala-Ala-Asn, Maleimide-(PEG)6-Ala-Ala-Asn, Maleimide-(PEG)8-Ala-Ala-Asn, Maleimide-(PEG)4-triazole-(PEG)3-sulfonamide, Maleimide-(PEG) 2- CH2CH2C(O)-Val-lys, maleimide-(PEG)4-triazole-(PEG)3-sulfonamide or Mal-(PEG)4-triazole-(PEG)3-disulfide.

[0068] In some embodiments, the linker L in the antibody-drug conjugate comprises: maleimide-(PEG)4-CH2C(O)-Gly-Gly-Phe-Gly, maleimide-(PEG)2-CH2CH2 C(O)-Gly-Gly-Phe-Gly, maleimide-(PEG)6-CH2C(O)-Gly-Gly-Phe-Gly-, maleimide-(CH2)5C(O)-Gly-Gly-Phe-Gly-, maleimide-C1-8 Alkylene-cycloalkyl-C(O)-NH(CH2CH2O)4CH2C(O)-Gly-Gly-Phe-Gly-, maleimide-(PEG)2-CH2C(O)-Gly-Gly-Phe-Gly-, maleimide-(PEG)2-CH2CH2C(O)-Val-Cit-, maleimide-(PEG)2-Gly-Gly-Phe-Gly-, maleimide-(PEG)2-CH2C(O)-Val-Cit-, maleimide-(PEG)4-CH2C(O)-Val-Cit- and maleimide-(PEG)6-CH2C(O)-Val-Cit-.

[0069] On the other hand, some embodiments provide the antibody-drug conjugate wherein Str is selected from a chemical moiety represented by the following formula:

[0070] where R 8 Selected from C 1-10 Alkylene, C 2-10 Alkenylene, (C 1-10 Alkylene)O-, N(R d )-(C 2-6 Alkylene)-N(R d ) and N(R d )-(C 2-6 alkylene); and each R d are independently H or C1-C6 alkyl.

[0071] In some embodiments, the antibody-drug conjugate is represented by a formula selected from the group consisting of:

[0072] where R 2 It is C 1-6 Alkylene, (C 1-6 Alkyl) NHC(NH)NH2 or (C 1-6 Alkylene)NHC(O)NH2;

[0073] where R 2 It is C 1-6 Alkyl, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 Alkylene)NHC(O)NH2;

[0074] where R 2 It is C 1-6 Alkyl, C 2-6 Alkenylene, (C 1-6Alkylene) NHC(NH)NH2 or (C 1-6 Alkylene)NHC(O)NH2;

[0075] where R 2 It is C 1-6 Alkyl, C 2-6 Alkenylene, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 Alkylene)NHC(O)NH2;

[0076] where R 2 It is C 1-6 Alkyl, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 alkylene)NHC(O)NH2, and R 5 and R 6 Together they form C 3-7 cycloalkyl ring;

[0077] where R 2 It is C 1-6 Alkyl, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 alkylene)NHC(O)NH2, and R 5 and R 6 Together they form C 3-7 cycloalkyl ring;

[0078] W, Str, and D are as described above.

[0079] In other embodiments, the present invention discloses an antibody-drug conjugate (ADC) represented by the following formula:

[0080] where R 2 Selected from C 1-6 Alkylene-NH2, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 Alkylene) NHC(O)NH2, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0081] where R 2 Selected from C 1-6 Alkylene-NH2, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6Alkylene) NHC(O)NH2, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0082] where R 2 It is C 1-6 Alkylene-NH2, (C 1- 6 alkylene) NHC(NH)NH2 or (C 1-6 alkylene)NHC(O)NH2, and R 5 and R 6 Formation C 3-7 Cycloalkyl ring, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0083] where R 2 It is C 1-6 Alkylene-NH2, (C 1-6 Alkylene) NHC(NH)NH2 or (C 1-6 alkylene)NHC(O)NH2, and R 5 and R 6 Formation C 3-7 Cycloalkyl ring, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0084] Y, R 3 、R 4 , Ab, and D are as described above.

[0085] On the other hand, some embodiments provide an antibody-drug conjugate (ADC) represented by the following formula:

[0086] where R 8 Selected from hydrogen, C 3-6 Cycloalkylalkyl or C 3-6 Cycloalkyl, preferably hydrogen; R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0087] where R 8 Selected from hydrogen, C 3-6 Cycloalkylalkyl or C 3-6 Cycloalkyl, preferably hydrogen; R 9 Selected from hydrogen, haloalkyl or C 3-6Cycloalkyl, preferably hydrogen; or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl; k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; p3 is selected from 0, 1 or 2;

[0088] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from an integer between 2 and 6;

[0089] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from an integer between 2 and 6;

[0090] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; p3 is selected from 0, 1 or 2;

[0091] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; and p3 is selected from 0, 1 or 2; k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8;

[0092] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; p3 is selected from 0, 1 or 2;

[0093] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8;

[0094] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; and p3 is selected from 0, 1 or 2;

[0095] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from 2, 4, 6 or 8;

[0096] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from 2, 4, 6 or 8;

[0097] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from 2, 4, 6 or 8;

[0098] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from 2, 4, 6 or 8;

[0099] k is selected from 1 to 10 and can be an integer or a decimal; p2 is selected from 2, 4, 6 or 8;

[0100] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; and p3 is selected from 0, 1 or 2;

[0101] k is selected from 1 to 10 and can be an integer or a decimal; p1 is selected from 2, 4, 6 or 8; and p3 is selected from 0, 1 or 2;

[0102] where R 8 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, k is selected from 1 to 10, which can be an integer or a decimal, and p2 is selected from an integer between 2 and 6;

[0103] where R 8 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl; k is selected from 1 to 10, which can be an integer or a decimal; p2 is selected from an integer between 2 and 6;

[0104] where R 8 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, k is selected from 1 to 10, which can be an integer or a decimal, p1 is selected from 2, 4, 6 or 8, and p3 is selected from 0, 1 or 2;

[0105] where R 8 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, R 9 Selected from hydrogen, haloalkyl or C 3-6 Cycloalkyl, preferably hydrogen, or R 8 and R 9 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, k is selected from 1 to 10 and can be an integer or a decimal, p1 is selected from 2, 4, 6 or 8, and p3 is selected from 0, 1 or 2.

[0106] In some embodiments, the anti-HER2 antibody drug conjugate is

[0107] k1 is 3 to 8, k2 is 3 to 8, and k1 and k2 are decimals or integers.

[0108] In some embodiments, the dosage of the compound of Formula IA or a pharmaceutically acceptable salt thereof is 0.01 mg / kg to 10.0 mg / kg. In alternative embodiments, the dosage of the compound of Formula IA or a pharmaceutically acceptable salt thereof is 0.01 mg / kg, 0.05 mg / kg, 0.08 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.3 mg / kg, 0.35 mg / kg, 0.4 mg / kg, 0.45 mg / kg, 0.5 mg / kg, 0.55 mg / kg, 0.6 mg / kg, 0.75 mg / kg, 0.85 mg / kg, 0.95 mg / kg, 100 mg / kg, 150 ... .6mg / kg, 0.65mg / kg, 0.7mg / kg, 0.75mg / kg, 0.8mg / kg, 0.85mg / kg, 0.9mg / kg, 0.95mg / kg, 1.0mg / kg, 1.25mg / kg, 1.5mg / kg, 1.75mg / kg, 2.0mg / kg, 2.2mg / kg, 2.4mg / kg, 2.6mg / kg, 2.8mg / kg, 3.0 mg / kg, 3.2mg / kg, 3.4mg / kg, 3.6mg / kg, 3.8mg / kg, 4.0mg / kg, 4.2mg / kg, 4.4mg / kg, 4.6mg / kg, 4. 8mg / kg, 5.0mg / kg, 5.2mg / kg, 5.4mg / kg, 5.6mg / kg, 5.8mg / kg, 6.0mg / kg, 6.2mg / kg, 6.4mg / kg, 6. In an optional embodiment, the dosage of the compound of formula IA or a pharmaceutically acceptable salt thereof is 0.08 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg or 1.5 mg / kg.

[0109] In some embodiments, the compound of Formula IA or a pharmaceutically acceptable salt thereof is administered once a week, once every two weeks, once every three weeks, or once every four weeks. In alternative embodiments, the administration frequency is once every two weeks or once every three weeks.

[0110] In an optional embodiment, the dosage of the compound of formula IA or a pharmaceutically acceptable salt thereof according to the present disclosure is 0.08 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg or 1.5 mg / kg, and the administration frequency is once every two weeks or once every three weeks.

[0111] The present disclosure also provides a method for treating anti-HER2 antibody-drug conjugate-resistant tumors, comprising administering to a subject a therapeutically effective amount of a compound of Formula IA or a pharmaceutically acceptable salt thereof.

[0112] The following are the variable region sequences of the heavy and light chains of Pertuzumab:

[0113] Light chain variable region

[0114] SEQ ID NO: 1

[0115] Heavy chain variable region

[0116] SEQ ID NO: 2

[0117] The following is the sequence of Pertuzumab:

[0118] light chain

[0119] SEQ ID NO.3

[0120] Heavy chain

[0121] SEQ ID NO.4

[0122] definition

[0123] In order to make the present disclosure more easily understood, certain technologies and sciences are specifically defined below. Unless otherwise explicitly defined in the present disclosure, all other technologies and sciences used in the present disclosure have the meanings commonly understood by those skilled in the art in the art to which the present disclosure belongs.

[0124] Unless the context clearly requires otherwise, throughout the specification and claims, the words "comprising," "having," "including," etc. should be construed to have an inclusive sense rather than an exclusive or exhaustive sense; that is, in the sense of "including but not limited to."

[0125] Antibody drug conjugates (ADCs) are antibodies that are linked to biologically active cytotoxins or small molecule drugs with cell-killing activity through a linker.

[0126] "Drug loading" or "drug loading" is also called the drug-to-antibody ratio (DAR), which is the average number of drugs conjugated to each antibody in the ADC. It can be, for example, in the range of 1 to 10 drugs conjugated to each antibody, and in certain embodiments, in the range of 1 to 8 drugs conjugated to each antibody, preferably 2-8, 2-7, 2-6, 2-5, 2-4, 3-4, 3-5, 5-6, 5-7, 5-8 and 6-8. Exemplary, the drug loading can be an average of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. The ADC formula of the present disclosure includes a collection of antibodies conjugated to drugs within the aforementioned ranges. In embodiments of the present disclosure, the drug loading can be represented as n. Drug loading can be determined by conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays and HPLC.

[0127] The term "linker unit" or "connection fragment" or "connection unit" refers to a chemical structure fragment or bond that is connected to an antibody or its antigen-binding fragment at one end and to a drug at the other end, and can also be connected to other linkers before being connected to the drug.

[0128] Linkers, including extenders, spacers, and amino acid units, can be synthesized by methods known in the art, such as those described in US20050238649A1. The linker can be a "cleavable linker" that facilitates release of the drug in the cell. For example, an acid-labile linker (e.g., a hydrazone), a protease-sensitive (e.g., a peptidase-sensitive) linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker can be used (Chari et al., Cancer Research 52:127-131 (1992); U.S. Patent No. 5,208,020).

[0129] The term "drug linker fragment" or "drug-linker fragment" refers to a fragment formed by linking a drug to a linker unit, which can be linked to an antibody through the other end of the linker unit.

[0130] Cytotoxic drug loading can be controlled by the following non-limiting methods, including:

[0131] (1) Control the molar ratio of the drug linker fragment and the monoclonal antibody,

[0132] (2) Control reaction time and temperature,

[0133] (3) Select different reaction reagents.

[0134] The three letter and one letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0135] The term "antibody" as used herein is used in the broadest sense and encompasses various antibody structures, including but not limited to full-length antibodies and antibody fragments (or antigen-binding fragments, or antigen-binding portions), as long as they exhibit the desired antigen-binding activity. Typically, a natural intact antibody consists of a tetrapeptide chain structure consisting of two identical heavy chains and two identical light chains connected by interchain disulfide bonds.

[0136] The engineered antibodies or antigen-binding fragments disclosed herein can be prepared and purified using conventional methods. For example, cDNA sequences encoding heavy and light chains can be cloned and recombined into GS expression vectors. The recombinant immunoglobulin expression vector can be stably transfected into CHO cells. As a more preferred existing technology, mammalian expression systems result in glycosylation of antibodies, particularly at the highly conserved N-terminal site in the Fc region. Positive clones are expanded in serum-free culture medium in a bioreactor to produce antibodies. The culture fluid that secretes antibodies can be purified using conventional techniques. For example, purification can be performed using an A or G Sepharose FF column containing an adjusted buffer. Non-specifically bound components are washed away. The bound antibodies are then eluted using a pH gradient method, and the antibody fragments are detected by SDS-PAGE and collected. The antibodies can be filtered and concentrated using conventional methods. Soluble mixtures and polymers can also be removed using conventional methods, such as molecular sieves and ion exchange. The resulting product should be immediately frozen, such as at -70°C, or freeze-dried.

[0137] As used herein, "Pertuzumab" refers to an antibody comprising the light chain and heavy chain variable region amino acid sequences in SEQ ID Nos. 1 and 2, respectively. If Pertuzumab is a complete antibody, it preferably comprises the light chain and heavy chain amino acid sequences in SEQ ID Nos. 3 and 4, respectively.

[0138] The numerical values ​​in this disclosure are instrumental measurements or calculated values ​​after instrumental measurement, and are subject to a certain degree of error. Generally speaking, a value within a reasonable error range of plus or minus 10% is within the reasonable error range. Of course, the context in which the numerical value is used must be considered. For example, the total impurity content, which is a value with an error variation of no more than plus or minus 10% after measurement, can be plus or minus 9%, plus or minus 8%, plus or minus 7%, plus or minus 6%, plus or minus 5%, plus or minus 4%, plus or minus 3%, plus or minus 2%, or plus or minus 1%, preferably plus or minus 5%.

[0139] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 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, 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, non-limiting examples of which 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, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0140] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic radical, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0141] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0142] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls. "Carbocycle" refers to the ring system within a cycloalkyl group.

[0143] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 6 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably piperidinyl and pyrrolidinyl. Polycyclic heterocyclic groups include spirocyclic, fused ring and bridged heterocyclic groups. "Heterocycle" refers to the ring system in the heterocyclic group.

[0144] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring. "Aromatic ring" refers to the ring system in an aryl group. Non-limiting examples of aryl groups include:

[0145] The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate, preferably phenyl.

[0146] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 12 members, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, pyrazolyl, pyrimidinyl or thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. "Heteroaromatic ring" refers to the ring system in a heteroaryl group. Non-limiting examples of heteroaryl groups include:

[0147] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0148] An "effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a subject may vary depending on factors such as the condition to be treated, the subject's overall health, the route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosage regimen that avoids significant side effects or toxic effects. The subject of the present disclosure may be an animal or a human subject.

[0149] The term "pharmaceutical composition" refers to a mixture containing one or more active ingredients described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0150] The term "pharmaceutically acceptable excipient" or "pharmaceutically acceptable vehicle" includes any material that, when combined with an active ingredient, allows the ingredient to retain biological activity and does not react with the subject's immune system. Examples include, but are not limited to, any standard pharmaceutical carrier, such as phosphate-buffered saline solution, water, emulsions such as oil / water emulsions, and various types of wetting agents. In some embodiments, the diluent for aerosol or parenteral administration is phosphate-buffered saline (PBS) or physiological (0.9%) saline. Compositions containing such carriers are formulated by well-known conventional methods (see, for example, Remington's Pharmaceutical Sciences, 18th edition, A. Gennaro, ed., Mack Publishing Co., Easton, PA, 1990; and R Remington, The Science and Practice of Pharmacy 20th edition Mack Publishing, 2000).

[0151] "Cancer," "cancerous," "proliferative disorder," and "tumor" as referred to in this disclosure are not mutually exclusive.

[0152] The terms "administer," "apply," and "treat" as applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with an animal, human, subject, cell, tissue, organ, or biological fluid, for example, for therapeutic, pharmacokinetics, diagnosis, research, and experimental procedures. Treatment of cells includes contact of an agent with a cell, as well as contact of an agent with a fluid, wherein the fluid is in contact with the cell. "Administer," "apply," and "treat" also mean the in vitro and ex vivo treatment of, for example, a cell, by an agent, a diagnostic, a binding composition, or by another cell. When applied to humans, veterinary medicine, or research subjects, they refer to therapeutic treatment, prophylactic or preventative measures, research, and diagnostic applications.

[0153] The term "treatment" means administering a therapeutic agent, such as a fusion protein or insulin analog comprising any of the present disclosure, to a subject who has, is suspected of having, or is predisposed to having one or more diabetes or hyperglycemia-related diseases or symptoms thereof, and for which the therapeutic agent is known to have a therapeutic effect. Typically, a therapeutic agent is administered in an amount effective to alleviate one or more disease symptoms in a treated subject or population, by preventing or delaying the onset of symptoms or complications, alleviating symptoms or complications, or eliminating the disease, condition, or disorder to any clinically measurable extent. The amount of a therapeutic agent effective to alleviate any specific disease symptom (also referred to as a "therapeutically effective amount") can vary according to a variety of factors, such as the disease state, age, and weight of the subject, and the ability of the drug to produce the desired therapeutic effect in the subject. Whether the disease symptoms have been alleviated can be evaluated by any clinical test method commonly used by a physician or other health care professional to evaluate the severity or progression of the symptoms. Although the embodiments of the present disclosure (e.g., methods of treatment or articles of manufacture) may not be effective in alleviating the symptoms of the target disease in a subject, they should alleviate the symptoms of the target disease in a statistically significant number of subjects as determined by any statistical test known in the art, such as Student's t-test, chi-square test, U test according to Mann and Whitney, Kruskal-Wallis test (H test), Jonckheere-Terpstra test, and Wilcoxon test. The patient to be treated is a mammal, and preferably a human.

[0154] The terms "prevent," "prevent," and "prevent" refer to reducing the risk or incidence of, or eliminating or slowing the progression of, one or more conditions, symptoms, complications, or disorders.

[0155] The terms "subject" and "patient" refer to mammals, particularly primates, and especially humans.

[0156] Overall survival (OS) is defined as the period from randomization to death from any cause. For subjects alive at the last follow-up, their OS was censored based on the last follow-up visit. For subjects lost to follow-up, their OS was censored based on the last confirmed survival time before loss to follow-up. OS for censored patients was defined as the time from randomization to censoring.

[0157] The objective response rate (ORR) refers to the proportion of patients whose tumor shrinkage reaches a certain level and is maintained for a certain period of time, including both complete response (CR) and partial response (PR). Objective response is assessed using the RECIST 1.1 criteria. Subjects must have measurable tumor lesions at baseline. Response assessment is based on RECIST 1.1 criteria, with complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD).

[0158] Disease control rate (DCR) refers to the percentage of confirmed complete remission, partial remission and stable disease (≥8 weeks) among patients who can be evaluated for efficacy.

[0159] Complete remission (CR): All target lesions disappear and the short diameter of all pathological lymph nodes (including target nodules and non-target nodules) must be reduced to <10mm.

[0160] Partial response (PR): The sum of the target lesion diameters decreased by at least 30% compared with the baseline level.

[0161] Disease progression (PD): The minimum value of the sum of all target lesion diameters measured during the entire experimental study is used as a reference, and the relative increase in the diameter sum is at least 20% (if the baseline measurement value is the minimum, the baseline value is used as a reference); in addition, the absolute value of the diameter sum must increase by at least 5 mm (the appearance of one or more new lesions is also considered as disease progression).

[0162] Stable disease (SD): The target lesion has not decreased to the level of PR, nor increased to the level of PD, but is somewhere in between. The minimum sum of the diameters can be used as a reference for research. BRIEF DESCRIPTION OF THE DRAWINGS

[0163] Figure 1 shows the effects of the test substances on the body weight of animals in the JIMT-1 nude mouse xenograft tumor model of breast cancer that is dual-resistant to Compound A and Compound B;

[0164] Figure 2 shows the effects of the test substances on the tumor volume of JIMT-1 breast cancer xenograft tumors in nude mice that are dual-resistant to Compound A and Compound B;

[0165] Figure 3 shows the effects of the test substances on the body weight of animals in the JIMT-1 nude mouse xenograft tumor model resistant to Compound B;

[0166] FIG4 shows the effects of the test substances on the tumor volume of compound B-resistant breast cancer JIMT-1 xenograft tumors in nude mice. DETAILED DESCRIPTION

[0167] The present disclosure is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present disclosure.

[0168] Example 1 Efficacy of Compound I in the JIMT-1 nude mouse xenograft tumor model of breast cancer dual-resistant to Compound A and Compound B

[0169] 1. Experimental Drugs

[0170] (1) Compound I, wherein n=4.1, Ab is Pertuzumab, and the preparation method can refer to PCT / CN2024 / 073229.

[0171] (2) Compound A, wherein k1=7.2, the preparation method can refer to WO2015115091.

[0172] (3) Compound B, wherein k2=6.0, the preparation method can refer to WO2020063676.

[0173] 2. Experimental animals: Female Balb / c Nude mice

[0174] 3. Test methods and steps

[0175] 1) Drug preparation

[0176] The compound A solution was stored at 4°C. Before use, 0.1 mL of the 20 mg / mL stock solution was aspirated, 1.9 mL of PBS was added, and the mixture was pipetted and mixed to obtain a homogeneous transparent liquid, thereby obtaining a working solution with a concentration of 1 mg / mL.

[0177] The compound B solution was stored at 4°C. Before use, 0.1 mL of the 20 mg / mL stock solution was aspirated and added to 1.9 mL of PBS. The mixture was pipetted and mixed to obtain a homogeneous transparent liquid. This was the working solution with a concentration of 1 mg / mL.

[0178] Compound I solution was stored at 4°C. Before use:

[0179] Pipette 0.025 mL of the 20 mg / mL stock solution, add 1.97 mL of PBS, and mix thoroughly with a pipette to obtain a uniform, transparent liquid. This is the working solution with a concentration of 0.25 mg / mL.

[0180] Pipette 0.05 mL of the 20 mg / mL stock solution, add 1.94 mL of PBS, and mix thoroughly with a pipette to obtain a homogeneous transparent liquid. This is the working solution with a concentration of 0.5 mg / mL.

[0181] Pipette 0.1 mL of the 20 mg / mL stock solution, add 1.9 mL of PBS, and mix thoroughly with a pipette to obtain a uniform transparent liquid, which is the working solution with a concentration of 1 mg / mL.

[0182] 2) Construction of mouse subcutaneous transplant tumor model

[0183] Tumor recovery: The JIMT-1 breast cancer tumor mass induced by compound A and compound B with dual drug resistance was removed from liquid nitrogen, quickly placed in a 37°C water bath for recovery, and cut into uniform tumor masses for later use.

[0184] Tumor inoculation: 70 female mice were inoculated with 10 mm tumor subcutaneously in the right upper limb of each experimental mouse. 3 Tumor tissue block. Regularly monitor tumor growth.

[0185] 3) Grouped medication

[0186] 35 days after the inoculation of JIMT-1 breast cancer cells with dual resistance to Compound A and Compound B, 42 tumors with a volume of 69.50-203.79 mm were selected. 3 Mice were randomly divided into six groups of seven animals each using the randomized block method. The day of grouping was designated D0. Dosing was performed twice according to the protocol in Table 1, on D0 and D14. Dissection was performed on D42. Tumor size and animal body weight were measured twice weekly during the experiment. Clinical symptoms were observed and recorded daily. All animals were euthanized after the final dose.

[0187] Table 1. Dosage regimen Note: ip intraperitoneal injection; “ / ” indicates not applicable

[0188] 4) Calculation

[0189] The calculation formula of tumor volume (TV) is: V = L × W 2 / 2. Where L and W represent the measured length and width of the tumor, respectively.

[0190] The relative tumor volume (RTV) was calculated as follows: RTV = Vt / V0, where V0 is the tumor volume measured at the time of dosing, and Vt is the tumor volume at the end of the treatment.

[0191] Relative tumor growth rate T / C (%) = (RTV of drug-treated group / RTV of vehicle group) × 100%.

[0192] Relative tumor change rate ΔT / ΔC (%) = (tumor volume at the end of the drug administration group - tumor volume at the beginning of the drug administration group) / (tumor volume at the end of the vehicle group - tumor volume at the beginning of the vehicle group) × 100%

[0193] Tumor growth inhibition rate TGI% = (1-ΔT / ΔC) × 100%

[0194] Body Weight (BW): The mice were weighed and monitored twice a week. The percentage body weight change (%BWC) was calculated as follows: %BWC = (BWc – BWi) / BWi × 100%, where c represents the current value and i represents the starting value.

[0195] 5) Test results

[0196] i. Effects of the test substance on the body weight of the JIMT-1 nude mouse xenograft tumor model with dual resistance to Compound A and Compound B

[0197] Compared to D0, the average body weight of mice in each group increased on D42 (the day of autopsy). The body weight changes in the control group, Compound A 10 mg / kg, Compound B 10 mg / kg, and Compound I 2.5 mg / kg, 5 mg / kg, and 10 mg / kg groups were 9.39%, 7.53%, 11.68%, 8.10%, 6.48%, and 3.76%, respectively. This indicates that the test drugs were safe. Detailed results are shown in Table 2 and Figure 1.

[0198] Table 2.

[0199] ii. Effects of the test substance on the tumor volume of the JIMT-1 nude mouse xenograft tumor model with dual resistance to Compound B and Compound A

[0200] At D42, the relative tumor growth inhibition rate (TGI%) of 10 mg / kg Compound A was 17.11%, and the TGI% of 10 mg / kg Compound B was -7.03%, indicating that the model was dual-resistant to Compound B and Compound A. Compound I at 2.5, 5, and 10 mg / kg significantly inhibited the growth of transplanted tumors (P < 0.01), with TGI% of 84.96%, 101.64%, and 113.11%, respectively.

[0201] The above results show that Compound I can significantly inhibit the growth of JIMT-1 breast cancer tumors that are dual-resistant to Compound B and Compound A in a dose-dependent manner. Detailed results are shown in Table 3 and Figure 2.

[0202] Table 3.

[0203] iii. Effect of the test substance on the tumor weight of JIMT-1 breast cancer xenograft tumors in nude mice with dual resistance to Compound B and Compound A

[0204] At D42, the relative tumor growth rate T / C% of 10 mg / kg compound A was 92.88%, and the relative tumor growth rate T / C% of 10 mg / kg compound B was 103.51%; the relative tumor growth rates T / C% of 2.5, 5, and 10 mg / kg compound I were 32.50%, 12.24%, and 3.92%, respectively.

[0205] These results demonstrate that the tumor inhibition rate calculated based on tumor weight on the day of dissection is consistent with that calculated based on tumor volume. Compound I significantly and dose-dependently inhibits the growth of JIMT-1 breast cancer xenografts in nude mice that are dual-resistant to both Compound B and Compound A. Detailed results are shown in Table 4.

[0206] Table 4.

[0207] 6) Experimental Conclusion

[0208] The results of this experiment showed that in JIMT-1 breast cancer cells that are dual-resistant to both Compound B and Compound A, Compound I can significantly inhibit the growth of JIMT-1 tumors that are dual-resistant to both Compound B and Compound A in a dose-dependent manner.

[0209] Example 2: Efficacy of Compound I and Compound A in the Compound B-resistant JIMT-1 nude mouse xenograft tumor model

[0210] 1. Experimental Drugs

[0211] (1) Compound I, wherein n=4.1.

[0212] (2) Compound A, wherein k1 = 7.2.

[0213] (3) Compound B, wherein k2 = 6.0.

[0214] 2. Experimental animals: Female Balb / c Nude mice

[0215] 3. Test methods and steps

[0216] 1) Drug preparation

[0217] The compound A solution was stored at 4°C. Before use, 0.1 mL of the 20 mg / mL stock solution was aspirated, 1.9 mL of PBS was added, and the mixture was pipetted and mixed to obtain a homogeneous transparent liquid, thereby obtaining a working solution with a concentration of 1 mg / mL.

[0218] The compound B solution was stored at 4°C. Before use, 0.1 mL of the 20 mg / mL stock solution was aspirated and added to 1.9 mL of PBS. The mixture was pipetted and mixed to obtain a homogeneous transparent liquid. This was the working solution with a concentration of 1 mg / mL.

[0219] Compound I solution was stored at 4°C. Before use:

[0220] Pipette 0.03 mL of the 20 mg / mL stock solution, add 1.97 mL of PBS, and mix thoroughly with a pipette to obtain a uniform, transparent liquid. This is the working solution with a concentration of 0.3 mg / mL.

[0221] Pipette 0.06 mL of the 20 mg / mL stock solution, add 1.94 mL of PBS, and mix thoroughly with a pipette to obtain a uniform, transparent liquid. This is the working solution with a concentration of 0.6 mg / mL.

[0222] Pipette 0.1 mL of the 20 mg / mL stock solution, add 1.9 mL of PBS, and mix thoroughly with a pipette to obtain a uniform transparent liquid, which is the working solution with a concentration of 1 mg / mL.

[0223] 2) Construction of mouse subcutaneous transplant tumor model

[0224] Tumor recovery: The JIMT-1 breast cancer tumor mass that had been induced to resist drug by two generations of compound B was removed from liquid nitrogen, quickly placed in a 37°C water bath for recovery, and cut into uniform tumor masses for later use.

[0225] Tumor inoculation: 100 female mice were inoculated subcutaneously with 10 mm tumor tissue in the right upper limb of each experimental mouse. 3 Tumor tissue block. Regularly monitor tumor growth.

[0226] 3) Grouped medication

[0227] 35 days after the inoculation of compound B-induced drug-resistant breast cancer JIMT-1, 42 tumors with a volume of 70.3-189.9 mm were selected. 3 Mice were randomly divided into 6 groups of 7 animals each using the randomized block method. The day of grouping was designated as D0. Dosing was performed twice as described in Table 5 on D0 and D14. Dissection was performed on D42. Tumor size and animal body weight were measured twice weekly during the experiment. Clinical symptoms were observed and recorded daily. All animals were euthanized after the final dose.

[0228] Table 5. Dosage regimen Note: ip intraperitoneal injection; “ / ” indicates not applicable

[0229] 4) Calculation

[0230] The calculation formula of tumor volume (TV) is: V = L × W 2 / 2. Where L and W represent the measured length and width of the tumor, respectively.

[0231] The relative tumor volume (RTV) was calculated as follows: RTV = Vt / V0, where V0 is the tumor volume measured at the time of dosing, and Vt is the tumor volume at the end of the treatment.

[0232] Relative tumor growth rate T / C (%) = (RTV of drug-treated group / RTV of vehicle group) × 100%.

[0233] Relative tumor change rate ΔT / ΔC (%) = (tumor volume at the end of the drug administration group - tumor volume at the beginning of the drug administration group) / (tumor volume at the end of the vehicle group - tumor volume at the beginning of the vehicle group) × 100%

[0234] Tumor growth inhibition rate TGI% = (1-ΔT / ΔC) × 100%

[0235] Body Weight (BW): The mice were weighed and monitored twice a week. The percentage body weight change (%BWC) was calculated as follows: %BWC = (BWc – BWi) / BWi × 100%, where c represents the current value and i represents the starting value.

[0236] 5) Test results

[0237] i. Effects of the test substance on the body weight of the JIMT-1 nude mouse xenograft tumor model induced by compound B

[0238] Compared to D0, the average body weight of mice in each group increased on D42 (the day of autopsy). The body weight changes in the control group, Compound B 10 mg / kg, Compound A 10 mg / kg, Compound I 3 mg / kg, 6 mg / kg, and 10 mg / kg groups were 7.06%, 7.68%, 8.46%, 7.84%, 3.74%, and 1.84%, respectively, indicating the good safety of the test drugs. Detailed results are shown in Table 6 and Figure 3.

[0239] Table 6

[0240] ii. Effects of the test substance on the tumor volume of the JIMT-1 nude mouse xenograft tumor model induced by compound B

[0241] At D42, 10 mg / kg of compound B had no inhibitory effect on the tumor (TGI% was 7.62%), indicating that the tumor had developed resistance to compound B; the TGI% of 10 mg / kg of compound A was 30.58%; 3, 6, and 10 mg / kg of compound I could significantly inhibit the growth of transplanted tumors (P<0.001), with TGI% of 109.13%, 115.70%, and 116.01%, respectively, and the efficacy was stronger than 10 mg / kg of compound A.

[0242] The above results show that Compound I can significantly inhibit the growth of JIMT-1 breast cancer xenograft tumors in nude mice that are resistant to Compound B. Detailed results are shown in Table 7 and Figure 4.

[0243] Table 7.

[0244] iii. Effects of the test substance on the weight of JIMT-1 breast cancer xenograft tumors in nude mice induced by compound B

[0245] At D42, the relative tumor growth rate T / C% of 10 mg / kg compound B was 93.53%; the relative tumor growth rate T / C% of 10 mg / kg compound A was 69.24%; the relative tumor growth rates T / C% of 3, 6, and 10 mg / kg compound I were 5.61%, 0.57%, and 0.16%, respectively.

[0246] These results demonstrate that the tumor inhibition rate calculated based on tumor weight on the day of dissection is consistent with that calculated based on tumor volume, demonstrating that Compound I significantly inhibits the growth of JIMT-1 breast cancer xenografts in nude mice that are resistant to Compound B. Detailed results are shown in Table 8.

[0247] Table 8

[0248] 6) Experimental Conclusion

[0249] The results of this experiment showed that in the JIMT-1 nude mouse xenograft tumor model of breast cancer that is resistant to Compound B, Compound I can inhibit tumor growth in a dose-dependent manner, and its anti-tumor activity is significantly better than that of Compound A. Tumor-bearing mice tolerated the test drugs well.

[0250] Example 3 A multicenter, open-label, phase I clinical study of the safety, tolerability, pharmacokinetics, and efficacy of Compound I in patients with unresectable or metastatic solid tumors expressing or mutating HER2

[0251] 1. Experimental Drugs

[0252] (1) Compound I, wherein n=4.1, lyophilized powder for injection, specification: 40 mg / bottle.

[0253] 2. Enrolled subjects

[0254] 1). Age 18-75 years old (inclusive):

[0255] 2) ECOG performance status score of 0 or 1;

[0256] 3) Dose escalation and PK expansion phase:

[0257] Patients with pathologically confirmed unresectable or metastatic solid tumors that express HER2 (IHC 1+ / 2+ / 3+) or are mutated and have not responded to standard treatment or have no effective standard treatment options.

[0258] Efficacy expansion stage:

[0259] Patients with pathologically confirmed, unresectable recurrent or metastatic solid tumors who have failed or have no standard treatment options. HER2 expression must be confirmed by the participating research center:

[0260] Cohort A: HER2-positive (IHC 3+ or ISH+) breast cancer: previously treated with trastuzumab and taxanes;

[0261] Cohort B: breast cancer with low HER2 expression (IHC 1+ and ISH-, or IHC 1+ and ISH undetectable, or IHC 2+ and ISH-);

[0262] Cohort C: Gastric cancer or gastroesophageal junction adenocarcinoma with HER2 overexpression (IHC 3+, or IHC 2+ and ISH+); previously received platinum-containing chemotherapy;

[0263] Cohort D: solid tumors other than the above tumor types (such as colorectal cancer, urothelial carcinoma, cholangiocarcinoma, non-small cell lung cancer, etc.) with HER2 expression (IHC 1+ / 2+ / 3+) or mutation (determined by second-generation sequencing or other appropriate analytical techniques).

[0264] 3. Dosage method

[0265] Eligible subjects were administered Compound I by intravenous infusion once every three weeks (Q3W), with a treatment cycle of 21 days. The doses were 0.08 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg, or 1.5 mg / kg.

[0266] Although specific embodiments of the present disclosure have been described above, those skilled in the art will appreciate that these are merely illustrative and that various changes or modifications may be made to these embodiments without departing from the principles and essence of the present disclosure. Therefore, the scope of protection of the present disclosure is defined by the appended claims.

Claims

1. Use of a compound of formula IA or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating tumor diseases resistant to anti-HER2 antibody-drug conjugates, in, Ab is Pertuzumab; n is 1 to 10, preferably 1 to 8, more preferably 3 to 5; The anti-HER2 antibody drug conjugate is selected from the compound represented by Formula II or a pharmaceutically acceptable salt thereof, Ab-(LD) k (II) wherein -D represents the residue of exitecan or its derivatives; L is a linker; Ab is anti-HER2 antibody; k is 1 to 10.

2. The method according to claim 1, wherein the tumor disease is selected from breast cancer, gastric cancer, colorectal cancer, urothelial carcinoma, bile duct cancer and lung cancer.

3. The use according to claim 1 or 2, wherein the tumor disease is selected from HER2-positive, HER2-low-expressing, HER2-overexpressing and HER2-mutated tumor diseases.

4. The method according to any one of claims 1 to 3, wherein -D is: in, A is -C(O)NH- or -O-; Every R 1a 、R 1b Each independently selected from hydrogen atom, C 1-6 alkyl, 3-6 membered cycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the alkyl, cycloalkyl, aryl or heteroaryl is optionally substituted with one or more substituents selected from C1-C6 alkyl, halogen, hydroxy, amino, oxo, 3-6 membered cycloalkyl, 6-10 membered aryl or C1-C6 alkoxy, or R 1a and R 1b Together with the carbon atom to which it is attached, it forms a 3-6 membered cycloalkyl group which is optionally substituted by one or more substituents selected from C1-C6 alkyl, halogen, hydroxy, amino, oxo or C1-C6 alkoxy; R 1c Selected from hydrogen atoms, deuterium atoms, C 1-6 Alkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the alkyl, aryl and heteroaryl are optionally substituted with one or more substituents selected from C1-C6 alkyl, halogen, hydroxyl, amino and oxo; n1 is an integer from 0 to 10.

5. The use according to any one of claims 1 to 4, wherein -D is:

6. The use according to any one of claims 1 to 5, wherein the anti-HER2 antibody drug conjugate is k1 is 3 to 8, and k2 is 3 to 8.

7. The use according to any one of claims 1 to 6, wherein the dosage of the compound represented by formula IA or a pharmaceutically acceptable salt thereof is 0.01 mg / kg to 10.0 mg / kg.

8. The use according to any one of claims 1 to 7, wherein the compound of formula IA or a pharmaceutically acceptable salt thereof is administered once a week, once every two weeks, once every three weeks or once every four weeks.

9. The use according to any one of claims 1 to 8, wherein the compound of formula IA or a pharmaceutically acceptable salt thereof is administered at a dosage of 0.08 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg or 1.5 mg / kg, and the administration frequency is once every two weeks or once every three weeks.

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