Methods of treating solid tumors

Abstract

Disclosed herein are methods of treating a solid tumor comprising administering to a patient having a solid tumor an effective amount of an antibody drug conjugate targeting TROP2.

Description

Methods for treating solid tumors Technical Field

[0001] The present invention relates to the field of drug treatment methods, and in particular to a method for using antibody-drug conjugates to treat solid tumors. Background Art

[0002] Targeted therapy for cancer, immunodeficiency, and infectious diseases is currently the focus of precision medicine. Over the years, many literatures have reported the use of cell surface receptor binding molecules as drug delivery vehicles to form conjugates with cytotoxic molecules for targeted delivery of cytotoxic molecules to attack various pathogenic cells (Allen, TM and Cullis, PR, 2004 Science, 303(5665), 1818-22, Hu, QY, et al. (2016), Chem Soc Rev 45(6): 1691-1719.).

[0003] Antibody-drug conjugates (ADCs) consist of three components: an antibody, a cytotoxic molecule, and a linker that connects the two (Thomas, A., et al. (2016), Lancet Oncol 17(6):e254-e262). Each of the three components has unique functions: the antibody needs to bind specifically to tumor cells, the cytotoxic molecule needs to have sufficient activity and broad spectrum against tumor cells, and the linker needs to have unique functionality, be stable in the blood circulation, and effectively release the cytotoxic molecule after reaching tumor cells (Chari, RV (2008), Acc Chem Res 41(1):98-107). Only when the three components are properly constructed can good clinical effects be achieved (Singh, SK, et al. (2015), Pharm Res 32(11):3541-3571; Hamilton, GS (2015), Biologicals 43(5):318-332).

[0004] TROP2 (Trophoblast cell surface antigen 2), also known as EGP-1, M1S1, or GA733-1, is a member of the TACSTD family, along with the epithelial cell adhesion molecule (EpCAM). It regulates downstream signaling through phosphorylation. TROP2 directly regulates the expression of the PARP1 enzyme, impairing the DNA repair function of normal cells. It also induces EMT, causing cancer cells to lose polarity and reduce adhesion between adjacent epithelial cells, ultimately leading to the metastasis and spread of cancer cells. TROP2 is overexpressed in a variety of cancer cells, including gastric cancer, colon cancer, papillary thyroid carcinoma, urothelial carcinoma, prostate cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, oral squamous cell carcinoma, ovarian epithelial carcinoma, and cervical cancer.

[0005] Summary of the Invention

[0006] The present invention provides a method for treating solid tumors, comprising administering an effective amount of an antibody-drug conjugate to a patient having a solid tumor; or using an antibody-drug conjugate in preparing a pharmaceutical composition for treating a solid tumor; or using an antibody-drug conjugate in treating a solid tumor; wherein the effective amount of the antibody-drug conjugate is administered approximately once every 1-8 weeks, with each administration dose being approximately 0.1-30 mg / kg, and the antibody-drug conjugate has a structure as shown in Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0007] in,

[0008] Abu is an antibody or antigen-binding unit thereof that binds to TROP2;

[0009] D is the drug;

[0010] M is Wherein * connects to Abu, ** connects to B, and R is selected from: -(CH2) r -、-(CHR m ) r -, C3-C8 carbocyclic group, -O-(CH2) r -, arylene, -(CH2) r -Arylene-, -Arylene-(CH2)r-, -(CH2) r -(C3-C8 carbocyclyl)-, -(C3-C8 carbocyclyl)-(CH2) r -, C3-C8 heterocyclic group, -(CH2) r -(C3-C8 heterocyclyl)-, -(C3-C8 heterocyclyl)-(CH2) r -、-(CH2) r C(O)NR m (CH2) r -、-(CH2CH2O) r -、-(CH2CH2O) r -CH2-, -(CH2) r C(O)NR m (CH2CH2O) r -、-(CH2) r C(O)NR m (CH2CH2O) r -CH2-, -(CH2CH2O) r C(O)NR m (CH2CH2O) r -、-(CH2CH2O)r C(O)NR m (CH2CH2O) r -CH2- and -(CH2CH2O) r C(O)NR m (CH2) r -; each R m R is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each r is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0011] B is or Where * connects to M, ** connects to L, and *** connects to G;

[0012] L is -(AA) i -(FF) f -, wherein AA is an amino acid or a polypeptide, and i is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0013] Each FF is independent Among them, each R F are independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; z is 0, 1, 2, 3 or 4, wherein * connects to AA and ** connects to D;

[0014] G is Where n is 1-24;

[0015] p is 1-10.

[0016] In one or more embodiments, the drug is an anticancer drug, a cytotoxic drug, a cell differentiation factor, a stem cell trophic factor, a steroid drug, a drug for treating an autoimmune disease, an anti-inflammatory drug, or a drug for treating an infectious disease.

[0017] In one or more embodiments, the drug is a tubulin inhibitor, a DNA damaging agent, or a DNA topoisomerase inhibitor.

[0018] In one or more embodiments, the tubulin inhibitor is selected from dolastatin, auristatin, and maytansine.

[0019] In one or more embodiments, the drug is an auristatin selected from monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF) or auristatin (AF).

[0020] In one or more embodiments, the drug is a DNA damaging agent, such as calicheamicins, duocarmycins, and anthramycin derivatives such as PBD (pyrrolobenzodiazepine).

[0021] In one or more embodiments, the drug is a DNA topoisomerase inhibitor or a salt thereof, such as irinotecan, irinotecan hydrochloride, isotecan derivatives, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, camptothecin derivatives SN-38, 22-hydroxyecliptazone, topotecan, lertotecan, belotecan, isotecan, homosilatecan, 6,8-dibromo-2 -methyl-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-(2E)-2-acrylamide, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(3-hydroxyphenylpropyl)-(E)-2-acrylamide, 12-β-D-glucopyranosyl-12,13-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide dihydrochloride, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide.

[0022] In one or more embodiments, the DNA topoisomerase inhibitor is camptothecin, 10-hydroxycamptothecin, topotecan, belotecan, irinotecan, 22-hydroxyelicholine or exitecan, or a salt thereof.

[0023] In one or more embodiments, the drug is

[0024] in

[0025] X 1 and X 2 Each independently is:

[0026] H,

[0027] hydroxyl groups,

[0028] C1-C6 alkyl,

[0029] C1-C6 alkyl substituted by one or more hydroxyl, halogen, nitro or cyano groups,

[0030] C2-C6 alkenyl,

[0031] C2-C6 alkynyl,

[0032] C1-C6 alkoxy,

[0033] C1-C6 aminoalkoxy,

[0034] halogen,

[0035] Nitro,

[0036] cyano group,

[0037] Thiol,

[0038] Alkylthio,

[0039] amino, amino substituted by an amino protecting group, C1-C6 aminoalkyl optionally substituted by an amino protecting group or a C1-C6 alkyl group at the amino moiety,

[0040] C1-C6 aminoalkylamino optionally substituted with an amino protecting group or a C1-C6 alkyl group at the amino moiety,

[0041] C1-C6 alkyl group connected to a heterocyclic ring, said heterocyclic ring being optionally substituted with one or more C1-C6 alkyl groups, C1-C6 alkoxy groups, amino groups, halogen groups, nitro groups or cyano groups,

[0042] C1-C6 alkylamino group connected to a heterocyclic ring, the heterocyclic ring is optionally substituted by C1-C6 alkyl, C1-C6 alkoxy, the amino group is optionally substituted by an amino protecting group, halogen, nitro, cyano or a protecting group,

[0043] An amino-substituted heterocyclic group, which is optionally substituted by a protecting group or one or more C1-C6 alkyl groups at the nitrogen atom of the heterocyclic ring or the amino group,

[0044] Heterocyclic amino group, which is optionally substituted by a protecting group or a C1-C6 alkyl group at the nitrogen atom of the heterocyclic ring portion or the amino portion,

[0045] a carbamoyl group optionally substituted with a carbamoyl protecting group or a C1-C6 alkyl group,

[0046] Morpholin-1-yl, or

[0047] Piperidin-1-yl;

[0048] X 3 is a C1-C6 alkyl group;

[0049] X 4 H, -(CH2) q -CH3、-(CHR n ) q-CH3, C3-C8 carbocyclyl, -O-(CH2) q -CH3, arylene-CH3, -(CH2) q -Arylene-CH3, -Arylene-(CH2) q -CH3, -(CH2) q -(C3-C8 carbocyclyl)-CH3, -(C3-C8 carbocyclyl)-(CH2) q -CH3, C3-C8 heterocyclic group, -(CH2) q -(C3-C8 heterocyclyl)-CH3, -(C3-C8 heterocyclyl)-(CH2) q -CH3, -(CH2) q C(O)NR n (CH2) q -CH3, -(CH2CH2O) q -CH3, -(CH2CH2O) q -CH2-CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH2-CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH2-CH3 or -(CH2CH2O) q C(O)NR n (CH2) q -CH3; each R n is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0050] **Connect with L;

[0051] y is 0, 1, or 2;

[0052] Y is O, S or CR 1 R 2 , where R 1 and R 2 Each independently is H or C1-C6 alkyl;

[0053] s and t are each independently 0, 1 or 2, but not both 0.

[0054] In one or more embodiments, X 4 is H or C1-C6 alkyl.

[0055] In one or more embodiments, the heterocycle is azetidine, niverazine, morpholine, pyrrolidine, piperidine, imidazole, thiazole, oxazole, or pyridine.

[0056] In one or more embodiments, the amino protecting group is formyl, acetyl, trityl, tert-butoxycarbonyl, benzyl or p-methoxybenzyloxycarbonyl.

[0057] In one or more embodiments, the drug is

[0058] where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; ** is connected to L.

[0059] In one or more embodiments, the drug is

[0060] where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; ** is connected to L.

[0061] In one or more embodiments, X 1 and X 2 Each is -CH3.

[0062] In one or more embodiments, X 1 and X 2 are each independently F, Cl, Br or I.

[0063] In one or more embodiments, X 1 and X 2 Each is F.

[0064] In one or more embodiments, X 1 and X 2 Each is independently -CH3, F or -OH.

[0065] In one or more embodiments, X 1 and X 2 Each is independently F or -CH3.

[0066] In one or more embodiments, X 1 -CH3 and X 2 For F.

[0067] In one or more embodiments, R is -(CH2)r-.

[0068] In one or more embodiments, R is -(CH2) r -, r is 1 or 5.

[0069] In one or more embodiments, R is -(CH2)-.

[0070] In one or more embodiments, each AA is independently selected from the following amino acid or peptide sequences: Val-Cit, Val-Lys, Phe-Lys, Lys-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp, Cit, Phe-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Leu-Ala-Leu, Ile-Ala-Leu, Val-Ala-Val, Ala-Leu-Ala-Leu, β-Ala-Leu-Ala-Leu, and Gly-Phe-Leu-Gly.

[0071] In one or more embodiments, i is 1.

[0072] In one or more embodiments, AA is Val-Cit, and i is 1.

[0073] In one or more embodiments, each FF is independently Among them, each R F are independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; wherein * connects to AA and ** connects to D.

[0074] In one or more embodiments, the halogen is F.

[0075] In one or more embodiments, each R F independently -CH3, F, -NO2 or -OCH3.

[0076] In one or more embodiments, z is 0.

[0077] In one or more embodiments, z is 1 or 2.

[0078] In one or more embodiments, f is 1.

[0079] In one or more embodiments, each FF is independently Where * connects AA and ** connects D.

[0080] In one or more embodiments, f is 1.

[0081] In one or more embodiments, FF is f is 1; * connects AA and ** connects D.

[0082] In one or more embodiments, L is Where * connects to B and ** connects to D.

[0083] In one or more embodiments, L is Where * connects to B and ** connects to D.

[0084] In one or more embodiments, G is n is 4-12.

[0085] In one or more embodiments, n is 4-8.

[0086] In one or more embodiments, n is 4.

[0087] In one or more embodiments, n is 8.

[0088] In one or more embodiments, p is 2-8.

[0089] In one or more embodiments, p is 4-8.

[0090] In one or more embodiments, p is 4-5.

[0091] In one or more embodiments, p is 6-8.

[0092] In one or more embodiments, p is 7-8.

[0093] In one or more embodiments, the Abu is the hRS9 antibody.

[0094] In one or more embodiments, the antibody drug conjugate has a structure as shown in Formula I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10 or I-11, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0095] in

[0096] Abu is an antibody or antigen-binding unit thereof that binds to TROP2;

[0097] R is selected from: -(CH2) r -、-(CHR m ) r -, C3-C8 carbocyclic group, -O-(CH2) r -, arylene, -(CH2) r -Arylene-, -Arylene-(CH2)r-, -(CH2) r -(C3-C8 carbocyclyl)-, -(C3-C8 carbocyclyl)-(CH2) r -, C3-C8 heterocyclic group, -(CH2) r -(C3-C8 heterocyclyl)-, -(C3-C8 heterocyclyl)-(CH2) r -、-(CH2) r C(O)NR m (CH2) r -、-(CH2CH2O) r -、-(CH2CH2O) r -CH2-, -(CH2) r C(O)NR m (CH2CH2O) r -、-(CH2) r C(O)NR m (CH2CH2O) r -CH2-, -(CH2CH2O) r C(O)NR m (CH2CH2O) r -、-(CH2CH2O) r C(O)NR m (CH2CH2O) r -CH2- and -(CH2CH2O) r C(O)NR m (CH2) r -; each R m R is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each r is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; or R is -(CH2) r -, or r is 1 or 5;

[0098] D is the drug;

[0099] n is an integer from 1 to 24; or n is 4 to 12;

[0100] p is 1-10, for example 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0101] In one or more embodiments, the drug is an anticancer drug, a cytotoxic drug, a cell differentiation factor, a stem cell trophic factor, a steroid drug, a drug for treating an autoimmune disease, an anti-inflammatory drug, or a drug for treating an infectious disease.

[0102] In one or more embodiments, the drug is a tubulin inhibitor, a DNA damaging agent, or a DNA topoisomerase inhibitor.

[0103] In one or more embodiments, the tubulin inhibitor is selected from dolastatin, auristatins, and maytansines.

[0104] In one or more embodiments, the drug is an auristatin, such as MMAE, MMAF, or AF.

[0105] In one or more embodiments, the drug is a DNA damaging agent, such as calicheamicins, duocarmycins, and anthramycin derivatives such as PBD (pyrrolobenzodiazepine).

[0106] In one or more embodiments, the drug is a DNA topoisomerase inhibitor or a salt thereof, such as irinotecan, irinotecan hydrochloride, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, camptothecin derivatives SN-38, 22-hydroxyecliptin, topotecan, lertotecan, belotecan, exitecan, homosilatecan, 6,8-dibromo-2-methyl-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-1,2-diol, 2-hydroxy ...2-diol, 2-hydroxy-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-1,2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2-diol, 2-hydroxy-2

[0015] The following are the esters of 1,2-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione: 1,2-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione, ...

[0107] In one or more embodiments, the DNA topoisomerase inhibitor is camptothecin, 10-hydroxycamptothecin, topotecan, belotecan, irinotecan, 22-hydroxyelicholine or exitecan, or a salt thereof.

[0108] In one or more embodiments, the drug is a tubulysin, a taxane derivative, a leptomycine derivative, CC-1065 and its analogs, an amatoxin, a spliceosome inhibitor, a benzodiazepine (PBD) dimer, doxorubicin, methotrexate, vincristine, vinblastine, daunorubicin, mitomycin C, melphalan, or a chlorambucil derivative.

[0109] In one or more embodiments, the drug is

[0110] in

[0111] X 1 and X 2 Each independently is:

[0112] H,

[0113] hydroxyl groups,

[0114] C1-C6 alkyl,

[0115] C1-C6 alkyl substituted by one or more hydroxyl, halogen, nitro or cyano groups,

[0116] C2-C6 alkenyl,

[0117] C2-C6 alkynyl,

[0118] C1-C6 alkoxy,

[0119] C1-C6 aminoalkoxy,

[0120] halogen,

[0121] Nitro,

[0122] cyano group,

[0123] Thiol,

[0124] Alkylthio,

[0125] amino, amino substituted by an amino protecting group, C1-C6 aminoalkyl optionally substituted by an amino protecting group or a C1-C6 alkyl group at the amino moiety,

[0126] C1-C6 aminoalkylamino optionally substituted with an amino protecting group or a C1-C6 alkyl group at the amino moiety,

[0127] C1-C6 alkyl group connected to a heterocyclic ring, said heterocyclic ring being optionally substituted with one or more C1-C6 alkyl groups, C1-C6 alkoxy groups, amino groups, halogen groups, nitro groups or cyano groups,

[0128] C1-C6 alkylamino group connected to a heterocyclic ring, the heterocyclic ring is optionally substituted by C1-C6 alkyl, C1-C6 alkoxy, the amino group is optionally substituted by an amino protecting group, halogen, nitro, cyano or a protecting group,

[0129] An amino-substituted heterocyclic group, which is optionally substituted by a protecting group or one or more C1-C6 alkyl groups at the nitrogen atom of the heterocyclic ring or the amino group,

[0130] Heterocyclic amino group, which is optionally substituted by a protecting group or a C1-C6 alkyl group at the nitrogen atom of the heterocyclic ring portion or the amino portion,

[0131] a carbamoyl group optionally substituted with a carbamoyl protecting group or a C1-C6 alkyl group,

[0132] Morpholin-1-yl, or

[0133] Piperidin-1-yl;

[0134] X 3 is a C1-C6 alkyl group;

[0135] X 4 H, -(CH2) q -CH3、-(CHR n ) q -CH3, C3-C8 carbocyclyl, -O-(CH2) q -CH3, arylene-CH3, -(CH2) q -Arylene-CH3, -Arylene-(CH2) q -CH3, -(CH2) q -(C3-C8 carbocyclyl)-CH3, -(C3-C8 carbocyclyl)-(CH2) q -CH3, C3-C8 heterocyclic group, -(CH2) q -(C3-C8 heterocyclyl)-CH3, -(C3-C8 heterocyclyl)-(CH2) q -CH3, -(CH2) q C(O)NR n (CH2) q -CH3, -(CH2CH2O) q -CH3, -(CH2CH2O) q -CH2-CH3, -(CH2) q C(O)NR n (CH2CH2O) q-CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH2-CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH2-CH3, or -(CH2CH2O) q C(O)NR n (CH2) q -CH3; each R n is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0136] ** is the connection point;

[0137] y is 0, 1, or 2;

[0138] Y is O, S or CR 1 R 2 , where R 1 and R 2 Each independently is H or C1-C6 alkyl;

[0139] s and t are each independently 0, 1 or 2, but not both 0.

[0140] In one or more embodiments, X 4 is H or C1-C6 alkyl.

[0141] In one or more embodiments, the heterocycle is azetidine, acetaldehyde, morpholine, pyrrolidine, piperidine, imidazole, thiazole, oxazole, or pyridine.

[0142] In one or more embodiments, the amino protecting group is formyl, acetyl, trityl, tert-butoxycarbonyl, benzyl or p-methoxybenzyloxycarbonyl.

[0143] In one or more embodiments, the drug is

[0144] where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; ** is the point of attachment.

[0145] In one or more embodiments, the drug is

[0146] where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; ** is the point of attachment.

[0147] In one or more embodiments, the C1-C6 alkyl group is -CH3.

[0148] In one or more embodiments, the halogen is F.

[0149] In one or more embodiments, X 1 and X 2 Each is independently -CH3, F or -OH.

[0150] In one or more embodiments, X 1 and X 2 Each is -CH3.

[0151] In one or more embodiments, X 1 and X 2 are each independently F, Cl, Br or I.

[0152] In one or more embodiments, X 1 and X 2 Each is F.

[0153] In one or more embodiments, X 1 and X 2 Each is independently F or -CH3.

[0154] In one or more embodiments, X 1 -CH3 and X 2 For F.

[0155] In one or more embodiments, R is -(CH2)r-.

[0156] In one or more embodiments, R is -(CH2)r-, and r is 1 or 5.

[0157] In one or more embodiments, R is -(CH2)-.

[0158] In one or more embodiments, n is 4-12.

[0159] In one or more embodiments, n is 4-8.

[0160] In one or more embodiments, n is 4.

[0161] In one or more embodiments, n is 8.

[0162] In one or more embodiments, p is 2-8.

[0163] In one or more embodiments, p is 4-8.

[0164] In one or more embodiments, p is 4-5.

[0165] In one or more embodiments, p is 6-8.

[0166] In one or more embodiments, p is 7-8.

[0167] In one or more embodiments, the antibody drug conjugate has a structure as shown in Formula I-12, I-13, I-14, I-15, I-16, I-17, I-18, I-19, I-20, I-21, I-22, I-23, I-24, or I-25, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0168] in

[0169] Abu is an antibody or antigen-binding unit thereof that binds to TROP2;

[0170] p is 1-10, for example 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0171] In one or more embodiments, p is 2-8.

[0172] In one or more embodiments, p is 4-8.

[0173] In one or more embodiments, p is 4-5.

[0174] In one or more embodiments, p is 6-8.

[0175] In one or more embodiments, p is 7-8.

[0176] In one or more embodiments, the Abu is an anti-Trop2 antibody.

[0177] In one or more embodiments, the anti-Trop2 antibody can specifically act on the Trop2 protein (trophoblast cell surface glycoprotein antigen 2).

[0178] In one or more embodiments, the anti-Trop2 antibody is a fully human monoclonal antibody.

[0179] In one or more embodiments, the anti-Trop2 antibody is a humanized monoclonal antibody.

[0180] In one or more embodiments, the anti-Trop2 antibody is an antibody disclosed in the following patent documents: WO2021147993A1 (such as PD3), CN101264325B (such as RS7, hRS7), CN105849126B (such as hTINA1-H1L1, hTINA1-H2L1, hTINA1-H2L2, hTINA1-H3L3), CN110903395A (such as M1, M2, M3), C N113896796A (such as 4D3, 7F11), US20130089872 (such as K5-70, K5-107, K5-116-2-1, T6-16, T5-86), US5840854A (such as BR110), US20130122020 (such as 3E9, 6G11, 7E6, 15E2, 18B1), US20120237518 (such as 77220, KM4097, KM4590).

[0181] In one or more embodiments, the anti-Trop2 antibodies are commercially available, including LS-C126418, LS-C178765, LS-C126416, LS-C126417 (LifeSpan BioSciences, Inc., Seattle, WA); 10428-MM01, 10428-MM02, 10428-R001, 10428-R030 (Sino Biological Inc., Beijing, China); MR54 (eBioscience, San Diego, CA); sc-376181, sc-376746, Santa Cruz Biotechnology (Santa Cruz, CA); MM0588-49D6 (Novus Biologicals, Littleton, CO); ab79976 and ab89928 (Cambridge, MA).

[0182] In one or more embodiments, the anti-Trop2 is the anti-Trop-2 antibodies 162-25.3 and 162-46.2 disclosed by Lipinski et al. (1981, Proc Natl. Acad Sci USA, 78: 5147-50) or the Pr1E11 anti-Trop-2 antibody disclosed by Ikeda et al. (2015, Biochem Biophys Res Comm 458: 877-82), which recognize a unique epitope on Trop-2.

[0183] In one or more embodiments, the anti-Trop2 antibody is hRS9 antibody, which comprises a heavy chain and a light chain having the sequences of SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

[0184] The amino acid sequences corresponding to the sequence numbers are shown in Table 1.

[0185] Table 1

[0186] In one or more embodiments, Abu is an antibody or an antigen-binding unit thereof that binds to TROP2, wherein the antibody or the antigen-binding unit thereof comprises a heavy chain shown in SEQ ID NO: 1 and a light chain shown in SEQ ID NO: 2.

[0187] In one or more embodiments, the ADC is ADC1 or a pharmaceutically acceptable salt or solvate thereof.

[0188] wherein p represents the average drug-antibody coupling ratio, which is about 3 to about 6. In one or more embodiments, p is about 4 to about 5. In one or more embodiments, the ADC is ADC2 or a pharmaceutically acceptable salt or solvate thereof.

[0189] wherein p represents the average drug-antibody conjugation ratio and is from about 3 to about 6. In one or more embodiments, p is from about 4 to about 5.

[0190] The present invention provides a pharmaceutical composition comprising an effective amount of the antibody-drug conjugate of the present invention and a pharmaceutically acceptable carrier.

[0191] Use of the antibody-drug conjugate or the pharmaceutical composition of the present invention in preparing a drug for treating solid tumors, wherein the drug comprises an effective amount of the antibody-drug conjugate of the present invention.

[0192] Examples of solid tumors include, but are not limited to, urothelial carcinoma, triple-negative breast cancer, ovarian cancer, uterine cancer, endometrial cancer, fallopian tube cancer, breast cancer, cervical cancer, gastric cancer, colorectal cancer, squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, liver cancer, bladder cancer, urethral cancer, hepatoma, breast cancer, colon cancer, rectal cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, glioblastoma, medulloblastoma, head and neck cancer, T-cell lymphoma, melanoma, liver cancer, Kaposi's sarcoma, hepatobiliary cancer, colorectal cancer, and large intestine cancer.

[0193] In one or more embodiments, the solid tumor is urothelial carcinoma, triple-negative breast cancer, gastric adenocarcinoma, esophageal cancer, hepatocellular carcinoma, non-small cell lung cancer, small cell lung cancer, epithelial ovarian cancer, cervical cancer, follicular thyroid carcinoma, glioblastoma multiforme, endometrial cancer, prostate cancer, head and neck squamous cell carcinoma, or renal clear cell carcinoma.

[0194] In one or more embodiments, the solid tumor is triple-negative breast cancer, non-small cell lung cancer, urothelial carcinoma, small cell lung cancer, or cervical cancer.

[0195] In one or more embodiments, the solid tumor is a TROP2-positive solid tumor.

[0196] In one or more embodiments, the solid tumor is a TROP2-positive advanced solid tumor.

[0197] In one or more embodiments, the dosage of each administration of the antibody drug conjugate is about 0.1-10 mg / kg. In one or more embodiments, the dosage of each administration of the antibody drug conjugate is about 0.1 mg / kg, about 0.8 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or a range (including endpoints) between any two of these values ​​or any value therein.

[0198] In one or more embodiments, the dosage of the antibody drug conjugate is about 5-3000 mg per administration. In one or more embodiments, the dosage of the antibody drug conjugate is about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg per administration. In one or more embodiments, the dose of the antibody drug conjugate administered per administration is about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints) or any value therein.

[0199] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose ranging from about 0.8 mg / kg to about 3.6 mg / kg.

[0200] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

[0201] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 2.7 mg / kg to about 3.3 mg / kg.

[0202] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 0.8 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg or about 3.6 mg / kg.

[0203] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg or about 3.6 mg / kg.

[0204] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 3.0 mg / kg.

[0205] In one or more embodiments, a treatment cycle is at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks or at least 7 weeks. In one or more embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks or 7 weeks, or the range (including endpoints) between any two values ​​of these numerical values ​​or any value therein. In one or more embodiments, approximately once a week, or approximately every 2 weeks, approximately every 3 weeks, approximately every 4 weeks, approximately every 5 weeks, approximately every 6 weeks, approximately every 7 weeks is administered 1 time. In one or more embodiments, approximately every 2 weeks is administered 1 time.

[0206] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2, 3, or 4 weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

[0207] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks.

[0208] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks.

[0209] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

[0210] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

[0211] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose ranging from about 2.7 mg / kg to about 3.3 mg / kg.

[0212] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose ranging from about 2.7 mg / kg to about 3.3 mg / kg.

[0213] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose being about 2.7 mg / kg.

[0214] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose being about 2.7 mg / kg.

[0215] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose being about 3 mg / kg.

[0216] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose being about 3 mg / kg.

[0217] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose being about 3.3 mg / kg.

[0218] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose being about 3.3 mg / kg.

[0219] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose being about 3.6 mg / kg.

[0220] In one or more embodiments, the effective amount of the antibody drug conjugate is administered once every three weeks, with each dose being about 3.6 mg / kg.

[0221] In one or more embodiments, the antibody drug conjugate is administered approximately once a week, and each dose is about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once a week, and each dose is about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once a week, and each dose is about 0.1 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or the range (including endpoints) between any two of these values ​​or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once a week, and each dose is about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once a week at a dose of about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once a week at a dose of about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, or about 2000 mg, or a range (including endpoints) between any two of these values ​​or any value therein.

[0222] In one or more embodiments, the antibody drug conjugate is administered approximately once every two weeks at a dose of about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every two weeks at a dose of about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every two weeks at a dose of about 0.5-4 mg / kg. In one or more embodiments, the antibody drug conjugate is administered about once every two weeks, with each dose being about 0.1 mg / kg, about 0.5 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or a range between any two of these values ​​(including the endpoints), or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once every two weeks at a dose of about 0.8 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, or about 7.2 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every two weeks at a dose of about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered about once every two weeks, with each dose being about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered about once every 2 weeks, with each dose being about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints), or any value therein.

[0223] In one or more embodiments, the antibody drug conjugate is administered approximately once every 3 weeks at a dose of about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 3 weeks at a dose of about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 3 weeks at a dose of about 0.5-4 mg / kg. In one or more embodiments, the antibody drug conjugate is administered about once every three weeks, with each dose being about 0.1 mg / kg, about 0.5 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or a range between any two of these values ​​(including the endpoints), or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once every three weeks at a dose of about 0.8 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, or about 7.2 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every three weeks at a dose of about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once every three weeks at a dose of about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered about once every three weeks, with each dose being about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints) or any value therein.

[0224] In one or more embodiments, the antibody drug conjugate is administered approximately once every 4 weeks at a dosage of about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 4 weeks at a dosage of about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 4 weeks at a dosage of about 0.1 mg / kg, about 0.5 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or any range (including endpoints) between any two of these values ​​or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once every four weeks at a dose of about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once every four weeks at a dose of about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered about once every four weeks, with each dose being about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, or about 1000 mg, or about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints), or any value therein.

[0225] In one or more embodiments, the antibody drug conjugate is administered approximately once every 5 weeks at a dosage of about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 5 weeks at a dosage of about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 5 weeks at a dosage of about 0.1 mg / kg, about 0.5 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or any range (including endpoints) between any two of these values ​​or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once every 5 weeks at a dose of about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 5 weeks at a dose of about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered about once every 5 weeks, with each dose being about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints), or any value therein.

[0226] In one or more embodiments, the antibody drug conjugate is administered approximately once every 6 weeks at a dosage of about 0.1-30 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 6 weeks at a dosage of about 0.1-10 mg / kg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 6 weeks at a dosage of about 0.1 mg / kg, about 0.5 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg, about 3.6 mg / kg, about 4.8 mg / kg, about 6.0 mg / kg, about 7.2 mg / kg, about 8.4 mg / kg, or about 10 mg / kg, or any range (including endpoints) between any two of these values ​​or any value therein. In one or more embodiments, the antibody drug conjugate is administered approximately once every 6 weeks at a dose of about 5-3000 mg. In one or more embodiments, the antibody drug conjugate is administered approximately once every 6 weeks at a dose of about 5-2000 mg, 5-1500 mg, 5-1000 mg, 5-800 mg, 5-600 mg, 48-432 mg, 48-216 mg, 72-504 mg, 100-2000 mg, 200-1000 mg, 300-800 mg, or 400-700 mg. In one or more embodiments, the antibody drug conjugate is administered about once every 6 weeks, with each dose being about 5 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, 500 mg, about 600 mg, about 700 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2500 mg, or about 3000 mg, or a range between any two of these values ​​(including the endpoints), or any value therein.

[0227] In one or more embodiments, the patient receives one treatment cycle. In one or more embodiments, the patient receives multiple (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26, or a range (including endpoints) between any two of these values ​​or any value therein) treatment cycles. In one or more embodiments, the patient receives treatment until the condition is alleviated and no longer requires treatment.

[0228] In one or more embodiments, in the methods for treating solid tumors provided by the present invention, the antibody drug conjugate can be administered by any convenient route, such as by infusion or bolus injection, absorbed through epithelial or skin mucosa (e.g., oral mucosa, rectal and intestinal mucosa, etc.). Thus, the pharmaceutical composition can be administered intravenously, subcutaneously, orally, rectally, parenterally, intracerebrally, intravaginally, intraperitoneally, topically (e.g., by powder, ointment, drops or transdermal patch), orally, or by oral or nasal spray. In one or more embodiments, the route of administration is intravenous infusion or subcutaneous injection.

[0229] In one or more embodiments, the antibody drug conjugate can be formulated into a pharmaceutical composition containing a single dose of the antibody drug conjugate as described above, such as a sterile isotonic aqueous solution, and administered to the patient in a form suitable for the selected route of administration, such as parenteral, intravenous (iv), intramuscular, topical or subcutaneous (sc). In one or more embodiments, the route of administration is intravenous infusion or subcutaneous injection.

[0230] In one or more embodiments, the methods or uses provided herein for treating solid tumors include administering an antibody drug conjugate in combination with one or more other therapies. Suitable other therapies include existing drugs (secondary drugs) for specific applications (e.g., cancer) and / or surgical therapies. For example, the antibody drug conjugate is administered in combination with one or more other chemotherapeutic or anti-tumor agents. Alternatively, the other therapeutic agent is radiation therapy. In one or more embodiments, the chemotherapeutic agent is a cell death inducer.

[0231] In one or more embodiments, the antibody drug conjugate and other agents are prepared as a single therapeutic composition, and the antibody drug conjugate and other agents are administered simultaneously. Alternatively, the antibody drug conjugate and other agents are independent of each other, for example, each prepared as an independent therapeutic composition, and the antibody drug conjugate and other agents are administered simultaneously, or the antibody drug conjugate and other agents are administered at different times during the treatment regimen. For example, the antibody drug conjugate is administered before the other agents, after the other agents, or in an alternating regimen. Herein, the antibody drug conjugate and other agents are administered in a single dose or multiple doses.

[0232] In one or more embodiments, in the methods for treating solid tumors provided by the present invention, the antibody drug conjugate and other agents can be administered by any convenient route, such as by infusion or bolus injection, absorbed by epithelial or mucocutaneous membranes (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be co-administered with other bioactive agents. Thus, the pharmaceutical composition can be administered intravenously, subcutaneously, orally, rectally, parenterally, intracerebrally, intravaginally, intraperitoneally, topically (e.g., by powder, ointment, drops or transdermal patch), orally, or by oral or nasal spray. In one or more embodiments, the route of administration is intravenous infusion or subcutaneous injection.

[0233] In one or more embodiments, the antibody drug conjugate and / or other co-administered pharmaceutical agent can be formulated into a pharmaceutical composition and administered to a patient in a form suitable for a selected route of administration, such as parenteral, intravenous (iv), intramuscular, topical, or subcutaneous (sc). In one or more embodiments, the route of administration is intravenous infusion or subcutaneous injection.

[0234] In one or more embodiments, the antibody drug conjugate, or the pharmaceutical composition comprising the antibody drug conjugate is administered by intravenous (iv) infusion (i.e., intravenous infusion). In one or more embodiments, the duration of intravenous infusion of the antibody drug conjugate, or the pharmaceutical composition comprising the antibody drug conjugate is about 15 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70 minutes, about 75 minutes, about 80 minutes, about 85 minutes, about 90 minutes, about 95 minutes, about 100 minutes, about 105 minutes, about 110 minutes, about 105 minutes, about 120 minutes, about 135 minutes, about 150 minutes, about 165 minutes, about 180 minutes, about 195 minutes, about 210 minutes, about 225 minutes, or about 240 minutes, or a range (including endpoints) between any two of these values, or any value therein. In one or more embodiments, the first intravenous infusion time of the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is ≥ 3 hours, and each subsequent intravenous infusion time is 1-2 hours.

[0235] In one or more embodiments, the intravenous infusion rate of the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is ≤2000 mg / hr. In one or more embodiments, the intravenous infusion rate of the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is ≤1000 mg / hr. In one or more embodiments, the intravenous infusion rate of the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is ≤500 mg / hr. In one or more embodiments, the rate of the first intravenous infusion of the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is ≤500 mg / hr, and the rate of each subsequent intravenous infusion is ≤1000 mg / hr. In one or more embodiments, when the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is intravenously infused, the rate from 0 to 15 minutes (initial rate) is ≤200 mg / hr, and starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 50-200 mg / hr. In one or more embodiments, when the antibody drug conjugate or the pharmaceutical composition comprising the antibody drug conjugate is first intravenously infused, the rate from 0 to 15 minutes (initial rate) is ≤50 mg / hr. Starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 50 mg / hr. For each subsequent injection, the rate from 0 to 15 minutes (initial rate) is 100-200 mg / hr. Starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 100-200 mg / hr. BRIEF DESCRIPTION OF THE DRAWINGS

[0236] Figure 1 shows the bystander effect of ADC1 and ADC2;

[0237] Figure 2 shows the changes in plasma concentration of ADC1 in rats;

[0238] Figure 3 shows the effect of ADC1 in inhibiting tumor growth;

[0239] FIG4 shows the tumor growth inhibitory effects of ADC1 and ADC3. DETAILED DESCRIPTION

[0240] "About" refers to the normal error range of the corresponding numerical value that is easily known to those skilled in the relevant art. In some embodiments, "about" mentioned herein refers to the described numerical value and its ±10%, ±5% or ±1% range.

[0241] "Treatment" refers to both therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent, slow, ameliorate or halt an undesirable physiological change or disorder, such as the progression of a disease, including but not limited to the following results, whether detectable or undetectable, relief of symptoms, reduction in the severity of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement, alleviation, reduction or elimination of the disease state (whether partial or complete), prolongation of life expectancy compared to that expected in the absence of treatment, etc. Patients in need of treatment include those already suffering from the disease or disorder, those susceptible to the disease or disorder, or those in need of prevention of the disease or disorder, and those who can or are expected to benefit from the administration of the antibodies or pharmaceutical compositions disclosed herein for detection, diagnostic procedures and / or treatment.

[0242] The term "TROP2-positive solid tumor" includes cancer cells with higher levels of TROP2 than normal tissue. Examples of TROP2-positive include, but are not limited to, TROP2-positive triple-negative breast cancer, TROP2-positive pancreatic cancer, urothelial carcinoma, non-small cell lung cancer, small cell lung cancer, glioblastoma, medulloblastoma, breast cancer, head and neck cancer, kidney cancer, ovarian cancer, gastric cancer, Kaposi's sarcoma, lung cancer, cervical cancer, colorectal cancer, esophageal cancer, oral squamous cell carcinoma, prostate cancer, thyroid cancer, bladder cancer, glioma, hepatobiliary tract cancer, colorectal cancer, T-cell lymphoma, colorectal cancer, prostate cancer, melanoma, and liver cancer.

[0243] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a drug, such as an antibody or ADC, that is sufficient to reduce or improve the severity and / or duration of a condition (e.g., cancer) or one or more symptoms thereof; prevent the progression of the condition; cause the condition to regress; prevent the recurrence, development, onset, or progression of one or more symptoms associated with the condition; detect the condition; or enhance or improve the preventive or therapeutic effect of another therapy (e.g., a prophylactic or therapeutic agent). For example, an effective amount of an antibody can inhibit tumor growth (e.g., inhibit an increase in tumor volume); reduce tumor growth (e.g., reduce tumor volume); reduce the number of cancer cells; and / or alleviate one or more symptoms associated with cancer to some extent. For example, an effective amount can improve progression-free survival (PFS), improve overall survival (OS), or reduce the likelihood of recurrence.

[0244] The terms "patient" and "subject" are used interchangeably and refer to any mammal in need of diagnosis, prognosis or treatment, including but not limited to humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cows, etc., particularly animals suffering from one or more conditions of solid tumors such as TROP2-positive solid tumors. In one or more embodiments, the patient is a human.

[0245] "Survival" means that the patient remains alive, including disease-free survival (DFS), progression-free survival (PFS) and overall survival (OS). Survival can be assessed by the Kaplan-Meier method, and any differences in survival can be calculated using the stratified log-rank test.

[0246] "Disease-free survival" refers to the time a patient has lived without their cancer returning since the start of treatment or since initial diagnosis. Events used in DFS analyses can include local, regional, and distant cancer recurrences, the development of secondary cancers, and death from any cause in patients without prior events.

[0247] "Prolonged survival" means PFS, DFS and / or OS in patients receiving increased treatment relative to untreated patients or relative to a control treatment regimen.

[0248] "Chemotherapy" or "chemotherapy" is the use of chemotherapeutic agents useful in the treatment of cancer.

[0249] “IC 50 ” stands for 50% inhibitory concentration, which is the concentration of drug or inhibitor required to inhibit a specified biological process by half.

[0250] As used herein, the term "administering" refers to administering a substance to achieve a therapeutic purpose (eg, treating a tumor).

[0251] As used herein, the term "tumor therapeutic drug" refers to an agent having the functional property of inhibiting the development or progression of a tumor in the human body, especially a malignant (cancerous) lesion such as carcinoma, sarcoma, lymphoma or leukemia. Inhibiting metastasis is a property of anti-tumor drugs in many cases.

[0252] MTD definition: The MTD (maximum tolerated dose) is defined as the highest dose level in a dose group at which DLT is observed in ≤ 1 / 6 subjects during the DLT evaluation period.

[0253] The PK (pharmacokinetic) parameters used in this article are defined as follows:

[0254] Single-dose PK parameter: C max 、T max 、T 1 / 2 , CL, Vd, Ke, MRT, AUC (0-τ) , AUC (0-∞) :

[0255] Multiple-dose PK parameters: C max,ss 、C avg,ss 、C min,ss , AUC (0-τ)ss , AUC (0-∞)ss 、Tmax,ss 、T 1 / 2,ss , CL, V ss , Ke, MRT, accumulation index (Rac), volatility index DF:

[0256] Objective response rate (ORR) refers to the proportion of patients whose tumor shrinks to a certain amount and remains for a certain period of time, including cases of CR (complete remission) and PR (partial remission).

[0257] Duration of response (DOR): DOR is defined as the time from the first objective response of the tumor to the first PD (disease progression) or death from any cause before PD, reflecting the duration of ORR.

[0258] Disease control rate (DCR): The proportion of patients whose tumors shrink or stabilize and remain stable for a certain period of time, including cases of CR, PR and SD (stable disease).

[0259] Progression-free survival (PFS): The time from the first dose to the occurrence of objective tumor progression or all-cause death (whichever occurs first).

[0260] Overall survival (OS) was calculated from the date of first dose to death from any cause. The cutoff date for subjects who were still alive at the time of analysis was the date of their last contact.

[0261] The term "antibody drug conjugate" or "ADC" is used interchangeably and refers to a binding protein (such as an antibody or its antigen binding unit) connected to one or more chemical drugs, which can optionally be a therapeutic agent or a cytotoxic agent. In some embodiments, the ADC includes an antibody, a drug (such as a cytotoxic drug) and a linker that enables the drug to be attached or coupled to the antibody. Non-limiting examples of drugs that can be included in an ADC are mitotic inhibitors, antitumor antibiotics, immunomodulators, vectors for gene therapy, alkylating agents, anti-angiogenic agents, antimetabolites, boron-containing agents, chemoprotectants, hormones, antihormonal agents, corticosteroids, photoactive therapeutic agents, oligonucleotides, radionuclide agents, topoisomerase inhibitors, kinase inhibitors (e.g., TEC-family kinase inhibitors and serine / threonine kinase inhibitors) and radiosensitizers.

[0262] The term "drug-antibody coupling ratio" or "DAR" refers to the number of drugs (e.g., exotecan) attached to an antibody of an ADC. The DAR of an ADC can range from 1 to 10, but higher loadings (e.g., 20) are possible depending on the number of attachment sites on the antibody. The term DAR can be used when referring to the number of drugs loaded onto a single antibody, or alternatively, when referring to the average or mean DAR of a group of ADCs. In some embodiments, the value is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. When considering the average number of drug-antibody bindings in a composition, that is, the average number of drug bindings for the antibody, or the average drug-antibody coupling ratio, the value is selected from about 0 to about 10, or about 2 to about 8. In some embodiments, the average drug-antibody coupling ratio is about 3 to about 6. In some embodiments, the average drug-antibody coupling ratio is about 4 to about 5. In other embodiments, the average drug-antibody coupling ratio is about 6 to about 8, or about 7 to about 8. DAR values ​​can be represented herein by p. The DAR value of an ADC can be measured using ultraviolet-visible absorption spectroscopy (UV-Vis), high-performance liquid chromatography-hydrophobic chromatography (HPLC-HIC), high-performance liquid chromatography-reverse phase chromatography (RP-HPLC), liquid chromatography-mass spectrometry (LC-MS), etc. These techniques are described in Ouyang, J. Methods Mol Biol, 2013, 1045: p. 275-83.

[0263] "Antibody" and "antigen-binding fragment" refer to polypeptides or polypeptide complexes that specifically recognize and bind to an antigen. An antibody can be a complete antibody, any antigen-binding fragment thereof, or a single chain thereof. Therefore, the term "antibody" includes any protein or peptide that contains at least a portion of an immunoglobulin molecule that has the biological activity of binding to an antigen. Antibodies and antigen-binding fragments include, but are not limited to, the complementarity determining regions (CDRs) of a heavy chain or light chain or its ligand-binding portion, a heavy chain variable region (VH), a light chain variable region (VL), a heavy chain constant region (CH), a light chain constant region (CL), a framework region (FR), or any portion thereof, or at least a portion of a binding protein. CDR regions include the CDR regions of the light chain variable region (VL CDR1-3) and the CDR regions of the heavy chain variable region (VH CDR1-3). An antibody or its antigen-binding unit can specifically recognize and bind to a polypeptide or polypeptide complex of one or more (e.g., two) antigens.

[0264] The term "antibody fragment" or "antigen-binding fragment" refers to a portion of an antibody. Antibody fragments of the present invention may be structured in forms similar to monospecific antibody fragments such as F(ab')2, F(ab)2, Fab', Fab, Fv, and scFv. Regardless of their structure, antibody fragments bind to the same antigen recognized by the intact antibody. The term "antibody fragment" includes aptamers, Spiegelmers, and diabodies. The term "antigen-binding fragment" also includes any synthetic or genetically engineered protein that functions as an antibody by binding to a specific antigen to form a complex.

[0265] "Polypeptide" refers to a molecule composed of two or more amino acid monomers linearly linked by an amide bond (also called a peptide bond), and may include dipeptides, tripeptides, tetrapeptides, oligopeptides, and the like.

[0266] "Amino acid" refers to an organic compound containing both an amino group and a carboxyl group, such as α-amino acids and β-amino acids. Amino acids include alanine (three-letter code: Ala, one-letter code: A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine ​​(Cys, C), glutamine (Gln, Q), glutamic acid (Glu, E), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), valine (Val, V), sarcosine (Sar), theanine (Thea), hydroxyproline (Hypro), hydroxylysine (Hylys), β-aminoisobutyric acid (β-AiBA), citrulline (Cit) and β-alanine (β-Ala), etc.

[0267] Those skilled in the art understand that when an amino acid or polypeptide is a component of a molecule (such as an antibody or ADC), the amino acid or polypeptide refers to an amino acid residue or a polypeptide residue (whether or not specified), that is, the remaining portion after some of its groups (such as a hydrogen atom of its amino group and / or a hydroxyl group of its carboxyl group) are lost due to the formation of a covalent bond (such as an amide bond) with the other parts of the molecule when it is combined with the other parts of the molecule.

[0268] Various substituents are defined as follows. In some cases, the number of carbon atoms in a substituent (for example, alkyl, alkenyl, alkynyl, alkoxy, aminoalkoxy, aminoalkyl, aminoalkylamino, alkylamino, heterocyclic radical, heterocyclic amino and aryl) is indicated by prefix "Cx-Cy" or "Cx-y", where x is the minimum value of carbon atoms and y is the maximum value of carbon atoms. Therefore, for example, "C1-C6 alkyl" refers to an alkyl group containing from 1 to 6 carbon atoms. If a substituent is described as "substituted by ... ", the hydrogen atoms on carbon or nitrogen are replaced by non-hydrogen groups. For example, a substituted alkyl substituent is an alkyl substituent in which at least one hydrogen atom on the alkyl group is replaced by a non-hydrogen group. For illustration, monofluoroalkyl is an alkyl group substituted by a fluoro group, and difluoroalkyl is an alkyl group substituted by two fluoro groups. It should be appreciated that if there is more than one substitution on a substituent, each substitution can be identical or different (unless otherwise indicated). If a substituent is described as being "optionally substituted with...", the substituent may be (1) unsubstituted or (2) substituted. Possible substituents include, but are not limited to, hydroxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 aminoalkoxy, halogen, nitro, cyano, mercapto, alkylthio, amino, C1-C6 aminoalkyl, C1-C6 aminoalkylamino, C1-C6 alkyl attached to a heterocycle, C1-C6 alkylamino attached to a heterocycle, heterocyclyl, amino-substituted heterocyclyl, heterocyclamino, carbamoyl, morpholin-1-yl, piperidin-1-yl, -(CH2) q -CH3、-(CHR n ) q -CH3, C3-C8 carbocyclyl, -O-(CH2) q -CH3, arylene-CH3, -(CH2) q -Arylene-CH3, -Arylene-(CH2) q -CH3, -(CH2) q -(C3-C8 carbocyclyl)-CH3, -(C3-C8 carbocyclyl)-(CH2) q -CH3, C3-C8 heterocyclic group, -(CH2) q -(C3-C8 heterocyclyl)-CH3, -(C3-C8 heterocyclyl)-(CH2) q -CH3, -(CH2) q C(O)NR n (CH2) q -CH3, -(CH2CH2O) q -CH3, -(CH2CH2O) q -CH2-CH3, -(CH2) q C(O)NR n(CH2CH2O) q -CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH2-CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH2-CH3, or -(CH2CH2O) q C(O)NR n (CH2) q -CH3; each R n is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0269] "Alkyl" refers to a saturated aliphatic hydrocarbon group, and the term includes straight and branched hydrocarbon groups. For example, C1-C20 alkyl, such as C1-C6 alkyl. C1-C20 alkyl refers to an alkyl group with 1 to 20 carbon atoms, for example, an alkyl group with 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms, 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms or 20 carbon atoms. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl etc. The alkyl group may be unsubstituted or substituted with one or more substituents, including but not limited to alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxyl, aryl, heteroaryl, amine, halogen, sulfonyl, sulfinyl, phosphonyl, and the like.

[0270] The term "alkenyl" by itself or as part of another substituent refers to an unsaturated branched, straight chain or cyclic alkyl group having at least one carbon-carbon double bond derived by the removal of a hydrogen atom from a single carbon atom of a parent alkene. Typical alkenyl groups include, but are not limited to, vinyl; propenyl (e.g., prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl, prop-2-en-2-yl, cycloprop-1-en-1-yl); cycloprop-2-en-1-yl; butenyl (e.g., but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, but-1,3-dien-1-yl, but-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobut-1,3-dien-1-yl, etc.), and the like.

[0271] The term "alkynyl" by itself or as part of another substituent refers to an unsaturated branched, straight-chain or cyclic alkyl group having at least one carbon-carbon triple bond, the triple bond being derived by removing a hydrogen atom from a single carbon atom of the parent alkyne. Typical alkynyl groups include, but are not limited to, ethynyl; propynyl (e.g., prop-1-yn-1-yl, prop-2-yn-1-yl, etc.); butynyl (e.g., but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.); and the like.

[0272] "Carbocyclyl" refers to a stable, non-aromatic, monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from 3 to 15 carbon atoms, for example, from 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) carbon atoms, and which is saturated or unsaturated and attached to the remainder of the molecule by a single bond. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decahydronaphthyl, etc. When specified, a carbocyclyl may be optionally substituted by one or more substituents independently selected from the group consisting of alkyl, halogen, haloalkyl, cyano, nitro, oxo, aryl, aralkyl, carbocyclyl, carbocyclylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, and heteroarylalkyl.

[0273] "Aryl" refers to an all-carbon monocyclic or all-carbon fused ring having a completely conjugated π electron system, typically having 5-14 carbon atoms, for example, 6, 10, 12, or 14 carbon atoms. An aryl group can be unsubstituted or substituted with one or more substituents including, but not limited to, alkyl, alkoxy, cyano, hydroxy, carboxyl, aryl, aralkyl, amine, halogen, sulfonyl, sulfinyl, and phosphonyl. Examples of unsubstituted aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl.

[0274] "Heterocyclyl" refers to a stable 3- to 18-membered aromatic or non-aromatic ring substituent consisting of 2 to 8 (e.g., 2, 3, 4, 5, 6, 7, or 8) carbon atoms and 1 to 6 (1, 2, 3, 4, 5, or 6) heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically stated, a heterocyclyl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocyclyl group can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl group can be partially or fully saturated. Examples of such heterocyclic groups include, but are not limited to, dioxolanyl, dioxinyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, piperrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, 1,2,4-thiadiazol-5(4H)-yl, tetrahydrofuranyl, trioxanyl, trithianyl, triazinanyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. When specifically stated in the specification, heterocyclyl can be optionally substituted by one or more substituents selected from the group consisting of alkyl, alkenyl, halogen, haloalkyl, cyano, oxo, thioxo, nitro, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl.

[0275] "Alkoxy" refers to the formula -O-(alkyl), where alkyl is an alkyl group as defined herein. Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy. Alkoxy groups may be substituted or unsubstituted.

[0276] "Halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0277] "Amino" refers to -NH2.

[0278] "Cyano" refers to -CN.

[0279] "Nitro" refers to -NO2.

[0280] "Hydroxyl" refers to -OH.

[0281] "Carboxyl" refers to -COOH.

[0282] "Mercapto" refers to -SH.

[0283] "Carbonyl" refers to C=O.

[0284] "Stereoisomers" refer to isomeric compounds that have the same order of attachment of atoms but different arrangements of atoms in space. Stereoisomers can have one or more stereocenters and each center can exist as R or S. Stereoisomers can also be cis-trans isomers. Stereoisomers of the compounds provided herein include any one of all diastereomeric, enantiomeric, and cis-trans isomeric forms thereof or suitable mixtures thereof.

[0285] Pharmaceutically acceptable salts include pharmaceutically acceptable salts of the compounds with a wide variety of organic and inorganic counterions well known in the art. Exemplary salts include, when the molecule contains an acidic functional group, organic or inorganic salts such as lithium, sodium, potassium, calcium, magnesium, ammonium, isopropylamine, trimethylamine, diethylamino, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine piperazine, piperidine, N-ethyl, piperidine, polyamine resins, and tetraalkylammonium salts; and when the molecule contains a basic functional group, organic or inorganic acid salts such as hydrochloride, hydrobromide, tartrate, methanesulfonate, acetate, maleate, and oxalate. Other non-limiting examples of acids include sulfuric acid, nitric acid, phosphoric acid, propionic acid, glycolic acid, pyruvic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid salicylic acid, etc. These salts can generally be prepared by conventional methods by reacting, for example, an appropriate acid or base with the compound. Solvates include hydrates.

[0286] Other chemical terms are used herein according to conventional usage in the art, as described in The McGraw-Hill Dictionary of Chemical Terms (Parker, S., ed., McGraw-Hill, San Francisco (1985)).

[0287] All publications and patents cited herein are incorporated by reference in their entirety for all purposes.

[0288] The antibody-drug conjugate of the present invention is a TROP2 antibody-drug conjugate (TROP2-ADC) formed by coupling an antibody or its antigen-binding unit that binds to TROP2 to a drug. Since the antibody-drug conjugate (ADC) can selectively deliver one or more drugs to the target tissue (e.g., a tumor expressing TROP2), the antibody-drug conjugate (ADC) can increase the therapeutic efficacy of the antibody in treating diseases (e.g., cancer). In one or more embodiments, the TROP2-ADC of the present invention has properties including but not limited to binding to TROP2 (e.g., human TROP2) in vitro, binding to cells expressing TROP2, high affinity, strong internalization ability, and reducing or inhibiting the growth of cancer cells or tumors.

[0289] In one or more embodiments, the TROP2-ADC is a structure as shown in Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt or solvate thereof.

[0290] wherein Abu, M, B, G, L, D, and p are as defined herein.

[0291] In one or more embodiments, Abu is the hRS9 antibody.

[0292] In one or more embodiments, the antibody-drug conjugates of the present invention have a significant bystander effect.

[0293] The present invention also provides methods for preparing intermediates and antibody-drug conjugates. The intermediates and antibody-drug conjugates of the present invention can be prepared using known formulations and methods. In one or more embodiments, the preparation methods are as follows.

[0294] Preparation method of the compound shown in formula II

[0295] in

[0296] M' is Where * connects B, R is selected from: -(CH2) r -、-(CHR m ) r -, C3-C8 carbocyclic group, -O-(CH2) r -, arylene, -(CH2) r -Arylene-, -Arylene-(CH2)r-, -(CH2) r -(C3-C8 carbocyclyl)-, -(C3-C8 carbocyclyl)-(CH2) r -, C3-C8 heterocyclic group, -(CH2) r -(C3-C8 heterocyclyl)-, -(C3-C8 heterocyclyl)-(CH2) r-、-(CH2) r C(O)NR m (CH2) r -、-(CH2CH2O) r -、-(CH2CH2O) r -CH2-, -(CH2) r C(O)NR m (CH2CH2O) r -、-(CH2) r C(O)NR m (CH2CH2O) r -CH2-, -(CH2CH2O) r C(O)NR m (CH2CH2O) r -、-(CH2CH2O) r C(O)NR m (CH2CH2O) r -CH2- and -(CH2CH2O) r C(O)NR m (CH2) r -; each R m R is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each r is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0297] L' is -(AA) i -(FF') f , wherein AA, i, and f are as described in Formula I; each FF' is independently Among them, each R F is independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; z is 0, 1, 2, 3 or 4; wherein * connects AA;

[0298] B and G are as described in Formula I.

[0299] Step 1: reacting a compound of formula 1 with a compound of formula 1' under alkaline conditions to obtain a compound of formula 2;

[0300] Step 2: Compounds of Formula 2 and Formula (AA) i -(FF 1 ) f In the presence of a condensing agent, react under alkaline conditions to obtain a compound of formula 3;

[0301] Step 3: removing the amino protecting group W1 of the compound of formula 3 to obtain the compound of formula 4;

[0302] Step 4: reacting the compound of formula 4 with the compound of formula 5 under alkaline conditions to obtain the compound of formula 6;

[0303] Step 5: The compound of formula 6 reacts with di(p-nitrobenzene) carbonate under alkaline conditions to obtain the compound of formula 7.

[0304] in

[0305] W1 is an amino protecting group, such as 9-fluorenylmethoxycarbonyl; W2 is a carboxylic acid active ester, such as succinimide ester.

[0306] Wherein, n, AA, R, i, and f are as described in Formula II above, and each FF 1 Independent Among them, each R F are independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; z is 0, 1, 2, 3 or 4; wherein * connects AA; each FF 2 Independent Among them, each R F is independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; z is 0, 1, 2, 3 or 4; wherein * connects AA.

[0307] In one or more embodiments, R F For F.

[0308] In one or more embodiments, z is 0.

[0309] In one or more embodiments, z is 1 or 2.

[0310] The above-mentioned alkaline conditions can be provided by reagents, which include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, diethylamine, N-methylmorpholine, pyridine, hexahydropyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide or potassium tert-butoxide; the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and lithium hydroxide.

[0311] The above condensing agent can be selected from N,N,N',N'-tetramethyl-O-(7-azabenzotriazole-1-yl) urea hexafluorophosphate, 4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholinium chloride, 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide, O-benzotriazole- N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate.

[0312] The preparation method of the intermediate shown in formula III

[0313] Wherein, B, G, L, and D are as described in Formula I, and M' is as described in Formula II.

[0314] Among them, n, AA, R, i, f, FF 2 , FF, and D are as described in this article.

[0315] The compound of formula 7 reacts with D in the presence of a condensing agent under alkaline conditions to obtain the compound of formula 8.

[0316] The above-mentioned alkaline conditions can be provided by reagents, which include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, diethylamine, N-methylmorpholine, pyridine, hexahydropyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide or potassium tert-butoxide; the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and lithium hydroxide.

[0317] The above condensing agent can be selected from N,N,N',N'-tetramethyl-O-(7-azabenzotriazole-1-yl) urea hexafluorophosphate, 4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholinium chloride, 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide, O-benzotriazole- N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate. Preparation method

[0318] wherein n, AA, R, i, f, FF, D, and Abu are as described herein.

[0319] The compound of formula 8 is coupled with Abu under weakly acidic conditions to obtain the compound of formula 9.

[0320] The above-mentioned weakly acidic conditions can be provided by reagents, which include organic acids and inorganic acids. The organic acids include but are not limited to acetic acid, benzoic acid, tartaric acid, oxalic acid, malic acid, citric acid, ascorbic acid, citric acid, citric acid, salicylic acid, caffeic acid, sorbic acid, quinic acid, oleanolic acid, succinic acid, chlorogenic acid, formic acid, and propionic acid. The inorganic acids include but are not limited to carbonic acid, nitrous acid, acetic acid, hypochlorous acid, hydrofluoric acid, sulfurous acid, hydrosulfuric acid, silicic acid, metasilicic acid, phosphoric acid, metaphosphoric acid, sodium bicarbonate, and sodium bisulfite.

[0321] In one or more embodiments, the ADCs of the present invention may be used in conjunction with other treatment or prevention regimens, including one or more ADCs of the present invention and one or more other therapeutic agents or methods. For combination therapy, the ADCs and other therapeutic agents may be administered simultaneously or separately. When administered separately, the ADCs of the present invention may be administered before or after administration of the other therapeutic agent.

[0322] Example

[0323] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources or known methods.

[0324] Example 1: Antibody Preparation

[0325] The following antibodies were prepared according to conventional methods. After constructing the vector, they were transiently transfected into eukaryotic cells such as HEK293 cells (Life Technologies Cat. No. 11625019), or stably transfected into CHO cells. Stable cell lines were selected and purified for expression.

[0326] Preparation and purification of hRS9 antibody: Anti-Trop2-specific monoclonal antibodies were produced in CHO cells according to the methods described in Wood et al., J Immunol. 145:3011 (1990). Expression vectors containing the antibody genes were constructed using conventional molecular biology methods. The amino acid sequences of the heavy and light chains of the recombinant humanized anti-Trop2 monoclonal antibody hRS9 are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

[0327] Example 2: Synthesis of Compound (1S,9S)-1-amino-9-ethyl-4,5-difluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione hydrochloride (D-1)

[0328] 1. Synthesis of N,N'-(3,4-difluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl)diethylamide

[0329] Under nitrogen, add potassium tert-butoxide solution (42 mL, 1 M) in tetrahydrofuran to a dry reaction flask with stirring and cool to 0-5°C. Slowly add N-(3,4-difluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (CAS No. 143655-49-6, 5 g, 21 mmol) dissolved in tetrahydrofuran (25 mL) dropwise to the reaction flask, followed by tert-butyl nitrite (4.32 g, 2 eq) dropwise (temperature controlled at 0-5°C). Heat to 15-20°C and stir for 2 hours. After the reaction is complete, cool to 0-5°C and add acetic acid (25 mL) and acetic anhydride (25 mL) dropwise (temperature controlled at no more than 10°C). Stir for 20 minutes. Maintain 5-10°C and add zinc powder (8 eq) according to the amount of N-(3,4-difluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide, and stir at 20-25°C for 1 hour. Filter, rinse the solid with ethyl acetate (50 mL), cool to 0-5°C, add 15% NaCO3 aqueous solution (50 mL) dropwise and wash three times, extract with ethyl acetate (25 mL), combine the organic phases, and wash with saturated NaCl aqueous solution. Add ethyl acetate (10 mL), stir at 40°C for 30 minutes, slowly reduce to 0-5°C and stir for 2 hours. Filter, wash the solid with ethyl acetate / petroleum ether (1 / 2, 10 mL). Dry in vacuo to obtain a gray powder (2.1 g, 33.7%). LC-MS: [M+H] + =297.

[0330] 2. Synthesis of N-(8-amino-5,6-difluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide

[0331] Add N,N'-(3,4-difluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl)diethylamide (500 mg, 1.68 mmol) to a 2M hydrochloric acid-ethanol solution (5 mL) and stir at 50°C for 4 h. After the reaction is complete, add water (7.5 mL), cool to 0-5°C, and add triethylamine (1.03 g) dropwise, stirring for 3 h. Filter and wash with 40% cold ethanol (3 mL) and water (3 mL), respectively. Dry in vacuo to obtain a gray powder (320 mg, 74.6%). LC-MS: [M+H] + =255.

[0332] 3. Synthesis of N-((9S)-9-ethyl-4,5-difluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3';4':6,7]indolizino[1,2-b]quinolin-1-yl)acetamide

[0333] Under nitrogen atmosphere, N-(8-amino-5,6-difluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (1.1 g, 1 eq), (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (1.15 g, 1 eq), and toluene (50 mL) were added to a reaction flask. The temperature was raised to reflux, stirred for 1 h, and then 4-methylbenzenesulfonic acid pyridine (100 mg) was added. Stirring and reflux were continued for 20 h. The temperature was lowered to room temperature and stirred for 1 h. The mixture was filtered and the solid was washed with acetone (10 mL) and cold ethanol (5 mL). The mixture was dried under vacuum to obtain a gray-brown powder (1.1 g, 53%). LC-MS: [M+H] + =482.

[0334] 4. Synthesis of (1S,9S)-1-amino-9-ethyl-4,5-difluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione hydrochloride

[0335] N-((9S)-9-ethyl-4,5-difluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)acetamide (1.1 g, 2.28 mmol) and 6M aqueous hydrochloric acid (44 mL) were added to a reaction flask and refluxed under nitrogen for 4 h. The solvent was removed by concentration and the product was purified by HPLC to obtain a white powder (200 mg, 18%). LC-MS: [M+H] + =440.

[0336] Example 3: Synthesis of 4-((30S,33S,36S)-30-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxo-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxo-27,32,35-triazaheptatriacontane-37-amino)benzyl(4-nitrophenyl)carbonate (CB07)

[0337] 1) Synthesis of (S)-30-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-26-oxo-2,5,8,11,14,17,20,23-octaoxa-27-aza-triacontane-31-oic acid (CB01).

[0338] Under nitrogen protection at 0-5°C, 8.14 g of N2-fluorenylmethoxycarbonyl-L-2,4-diaminobutyric acid (CB00-2) was dissolved in 40 mL of dimethylformamide (DMF), and 10 g of 4,7,10,13,16,19,22,25-octaoxacohexacanoic acid-N-succinimide ester (CB00-3) and 10 mL of DMF were added. 6.5 mL of DIPEA was added dropwise at 0-5°C. After 1 hour of addition, the mixture was stirred at room temperature for 4 hours. After the reaction was completed, DMF was removed under reduced pressure, and 14.2 g of CB01 was obtained as a light yellow oily liquid by silica gel column chromatography (dichloromethane and methanol in a volume ratio of 20:1 as the elution solvent).

[0339] 2) Synthesis of (9H-fluoren-9-yl)methyl [(S)-1-[[(S)-1-[[4-(hydroxymethyl)phenyl]amino]-1-oxo-5-ureidopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamate (CB02).

[0340] Under nitrogen at room temperature, 11 g of (S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutyrylamino)-5-ureidopentanic acid (CB00-4) and 5.5 g of p-aminobenzyl alcohol (CB00-5) were dissolved in 400 mL of dichloromethane and 200 mL of methanol. Under mechanical stirring, 17 g of 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) was added portionwise. The mixture was reacted in the dark for 15 h. After completion of the reaction, the solvent was removed under reduced pressure to obtain a paste-like solid. Silica gel column chromatography (dichloromethane:methanol, volume ratio 20:1) was used as the eluting solvent to obtain 11.9 g of an off-white solid.

[0341] 3) Synthesis of (S)-2-((S)-2-amino-3-methylbutyramido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide (CB03).

[0342] Under nitrogen protection at room temperature, 300 mL of acetonitrile was added to 11.9 g of [(S)-1-[[(S)-1-[[4-(hydroxymethyl)phenyl]amino]-1-oxo-5-ureidopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamic acid (9H-fluorene-9-yl)methyl ester (CB02), and 18 mL of piperidine was added dropwise with stirring. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 h. After completion of the reaction, the solvent and piperidine were removed by distillation under reduced pressure, and 7.5 g of CB03 was obtained as a white solid by silica gel column chromatography (dichloromethane and methanol volume ratio 20:1 as elution solvent).

[0343] 4) Synthesis of (9H-fluoren-9-yl)methyl ((30S,33S,36S)-41-amino-36-((4-(hydroxymethyl)phenyl)carbamoyl)-33-isopropyl-26,31,34,41-tetraoxo-2,5,8,11,14,17,20,23-octaoxa-27,32,35,40-tetraaza-30-yl)carbamate (CB04).

[0344] Under a nitrogen atmosphere at 0°C, 14.2 g of (S)-30-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-26-oxo-2,5,8,11,14,17,20,23-octaoxa-27-aza-triacontane-31-oic acid (CB01) was dissolved in 100 mL of DMF, and 11 g of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (HATU) was added portionwise. After stirring for 30 min, 7.5 g of (S)-2-((S)-2-amino-3-methylbutanamido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide (CB03) was added, and the reaction was maintained at 0°C for 2.5 h. After the reaction was completed, the solvent was distilled off under reduced pressure, and 9.66 g of solid CB04 was obtained by silica gel column chromatography (dichloromethane and methanol in a volume ratio of 10:1 as elution solvent).

[0345] 5) Synthesis of N-((S)-3-amino-4(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxyl)-2,5,8,11,14,17,20,23-octaoxahexacosa-26-amide (CB05).

[0346] Under nitrogen protection at room temperature, 9.66g of (9H-fluoren-9-yl)methyl ((30S,33S,36S)-41-amino-36-((4-(hydroxymethyl)phenyl)carbamoyl)-33-isopropyl-26,31,34,41-tetrahydro-2,5,8,11,14,17,20,23-octaoxa-27,32,35,40-tetraaza-30-yl)carbamate (CB04) was dissolved in 50mL DMF, 12mL diethylamine was added, and the reaction was stirred for 1.5h. After the reaction was completed, the solvent was distilled off under reduced pressure, and 7.7g CB05 was obtained as a light yellow solid by silica gel column chromatography (dichloromethane and methanol volume ratio 7.5:1 as elution solvent).

[0347] 6) Synthesis of N-((S)-3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-4-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-butanone-2-yl)amino)-4-oxo)-2,5,8,11,14,17,20,23-octaoxahexacosa-26-amide (CB06).

[0348] 7.7 g of N-((S)-3-amino-4(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxyl)-2,5,8,11,14,17,20,23-octaoxahexacosa-26-amide (CB05) was dissolved in 40 mL of DMF. Maleimidoacetic acid succinimidyl ester (CB00-1) was added portionwise under a nitrogen atmosphere at 0-5° C., and the reaction was maintained at 0-5° C. for 4 h. After completion of the reaction, the solvent was distilled off under reduced pressure, and 9.5 g of CB06 was obtained as a light yellow solid by silica gel column chromatography (dichloromethane and methanol in a volume ratio of 10:1 as the elution solvent).

[0349] 7) Synthesis of 4-((30S,33S,36S)-30-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxo-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxo-27,32,35-triazaheptatriacontane-37-amino)benzyl(4-nitrophenyl)carbonate (CB07).

[0350] 9.5 g of N-((S)-3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-4-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-butanone-2-yl)amino)-4-oxo)-2,5,8,11,14,17,20,23-octaoxahexacosanetan-26-carboxamide (CB06) was dissolved in 50 mL of DMF. Under a nitrogen atmosphere at 0°C, 14.0 g of di(p-nitrobenzene) carbonate ((PNP)2CO) was added. After dissolution, 8.2 mL of N,N-diisopropylethylamine (DIPEA) was added, and the reaction was maintained at 0°C for 4 h. After the reaction was completed, the solvent was distilled off under reduced pressure, and 2.6 g of CB07 was obtained as a white solid by silica gel column chromatography (dichloromethane and methanol in a volume ratio of 8:1 as the elution solvent).

[0351] Example 4: Synthesis of 4-((18S,21S,24S)-18-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-21-isopropyl-14,19,22-trioxo-24-(3-ureidopropyl)-2,5,8,11-tetraoxo-15,20,23-triazapentacosane-25-amino)benzyl(4-nitrophenyl)carbonate (CB14)

[0352] CB14 was prepared by referring to the synthesis of CB07 in Example 5, except that 4,7,10,13,16,19,22,25-octaoxahexacosadecanoic acid-N-succinimidyl ester was replaced with 4,7,10,13-tetraoxatetradecanoic acid-N-succinimidyl ester to obtain CB14 as a white solid.

[0353] Example 5: Synthesis of Intermediate (CB07-Exatecan)

[0354] Synthesis of 4-(30S,33S,36S)-30-(2-(2,5-dioxa-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxa-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxa-27,32,35-triazaheptatriaminoaconit-37-yl)benzoyl(1s,9s)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-carbamate.

[0355] 4-((30S,33S,36S)-30-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxo-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxo-27,32,35-triazatriacontane-37-amino)benzyl(4-nitrophenyl)carbonate (CB07) (2.6 g, 2.21 mmol) and N,N-dimethylformamide (23 mL) were added to reaction flask R1, stirred under nitrogen protection and cooled to 0-5°C. Meanwhile, (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione mesylate (Ixitecan mesylate, 0.98 g, 1.84 mmol, Advanced ChemBlocks) and N,N-dimethylformamide (5 mL) were added to another reaction flask R2. Triethylamine (230 mg, 2.27 mmol) was added dropwise at 0-5°C and stirred until completely dissolved. The solution in reaction flask R2 was added dropwise to reaction flask R1. After washing reaction flask R2 with N,N-dimethylformamide (2 mL), the washing solution was added to reaction flask R1. 1-Hydroxybenzotriazole (497 mg, 3.68 mmol) and pyridine (1.45 g, 18.4 mmol) were weighed and added to reaction flask R1. Stir at 0-5°C for 10 min, then warm to room temperature and stir for 5.5 h. After the reaction was complete, the solvent was removed by concentrating under reduced pressure at 35°C. Purification by preparative high-performance liquid chromatography (prep-HPLC) and lyophilization yielded a white powder (1.6 g, 59%). LC-MS: [1 / 2M+H] + =737.

[0356] Example 6: Synthesis of intermediate (CB07-D-1)

[0357] Synthesis of 4-(30S,33S,36S)-30-(2-(2,5-dioxa-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxa-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxa-27,32,35-triazaheptatriaminoaconit-37-yl)benzoyl (1S,9S)-9-ethyl-4,5-difluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-carbamate.

[0358] 4-((30S,33S,36S)-30-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-33-isopropyl-26,31,34-trioxo-36-(3-ureidopropyl)-2,5,8,11,14,17,20,23-octaoxo-27,32,35-triazaheptatriacontane-37-amino)benzyl(4-nitrophenyl)carbonate (CB07) (220 mg, 0.189 mmol) and N,N-dimethylformamide (5 ml) were added to reaction flask R1, stirred under nitrogen protection and cooled to 0-5°C. Meanwhile, (1S,9S)-1-amino-9-ethyl-4,5-difluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13Hbenzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione hydrochloride (D-1) (90 mg, 0.189 mmol) and N,N-dimethylformamide (5 mL) were added to another reaction flask R2. Three drops of triethylamine were added dropwise at 0-5°C and stirred until completely dissolved. The solution in reaction flask R2 was added dropwise to reaction flask R1. 1-Hydroxybenzotriazole (60 mg, 0.44 mmol) and pyridine (0.5 mL) were then weighed and added to reaction flask R1. Stirring was continued at 0-5°C for 10 minutes. After the mixture was heated to room temperature and stirred for 3 hours, the solvent was removed under reduced pressure. Purify by preparative high performance liquid chromatography (prep-HPLC) and freeze-dry to obtain a white powder (55 mg, 20%). LC-MS: [1 / 2M+H] + =739.

[0359] Example 7: Synthesis of Intermediate (CB14-Exatecan)

[0360] Synthesis of 4-(18S,21S,24S)-18-(2-(2,5-dioxane-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-21-isopropyl-14,19,22-trioxo-24-(3-ureidopropyl)-2,5,8,11-tetraoxo-15,20,23-triazapentacosan-25-amino)-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-carbamate.

[0361] Similarly, 4-((18S,21S,24S)-18-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)-21-isopropyl-14,19,22-trioxo-24-(3-ureidopropyl)-2,5,8,11-tetraoxo-15,20,23-triazapentacosan-25-amino)benzyl(4-nitrophenyl)carbonate (190 mg, 0.19 mmol) and N,N-dimethylformamide (23 mL) were added to reaction flask R1, and the mixture was stirred under nitrogen and cooled to 0-5°C. Meanwhile, (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione mesylate (101 mg, 0.19 mmol; Advanced ChemBlocks) and N,N-dimethylformamide (5 mL) were added to another reaction flask R2. Three drops of triethylamine were added dropwise at 0-5°C and stirred until completely dissolved. The solution in reaction flask R2 was added dropwise to reaction flask R1. After washing reaction flask R2 with N,N-dimethylformamide (1 mL), the washing solution was added to reaction flask R1. 1-Hydroxybenzotriazole (60 mg, 0.44 mmol) and pyridine (0.5 mL) were weighed and added to reaction flask R1. Stir at 0-5°C for 10 min, then warm to room temperature and stir for 5.5 h. After the reaction was complete, the solvent was removed by concentrating under reduced pressure at 35°C. Purification was performed by preparative high-performance liquid chromatography (prep-HPLC) and lyophilization was performed to obtain a white powder.

[0362] Example 8: Synthesis of ADC1

[0363] Based on the amount of antibody, 2.7 molar equivalents of TCEP were added to the hRS9 antibody, and the pH was adjusted to 7 with 1 M Tris base. The solution was incubated at 25°C for 2 hours to reduce the concentration. TCEP was removed by ultrafiltration using 10 mM succinic acid. The thiol concentration of the antibody was determined by measuring the absorbance at 412 nm of the reaction product of the thiol group with DTNB (5,5'-dithiobis(2-nitrobenzoic acid), Aldrich).

[0364] During the coupling reaction, 6 molar equivalents of CB07-Exatecan were added based on the amount of antibody. After stirring at 25°C for 2 hours, 0.1M acetylcysteine ​​was added to a final concentration of 2 mM and the reaction was terminated by stirring for 15 minutes. The reaction mixture was filtered through a 0.22 μm filter and eluted with 10 mM succinic acid through Sephadex G-25 resin.

[0365] The protein concentration of ADC1 was determined to be 8.46 mg / mL, and the DAR, i.e., p, was 4.2 as detected by RP-HPLC.

[0366] Example 9: Synthesis of ADC2

[0367] Based on the amount of antibody, 2.7 molar equivalents of TCEP were added to the hRS9 antibody, and the pH was adjusted to 7 with 1 M Tris base. The solution was incubated at 25°C for 2 hours to reduce the concentration. TCEP was removed by ultrafiltration using 10 mM succinic acid. The thiol concentration of the antibody was determined by measuring the absorbance at 412 nm of the reaction product of the thiol group with DTNB (5,5'-dithiobis(2-nitrobenzoic acid), Aldrich).

[0368] During the coupling reaction, 6 molar equivalents of CB07-D-1 were added based on the amount of antibody. After stirring at 25°C for 2 hours, 0.1M acetylcysteine ​​was added to a final concentration of 2 mM and the reaction was terminated by stirring for 15 minutes. The reaction mixture was filtered through a 0.22 μm filter and eluted with 10 mM succinic acid through Sephadex G-25 resin.

[0369] The protein concentration of ADC2 was determined to be 6.5 mg / mL, and the DAR, i.e., p, was 4.8 as detected by RP-HPLC.

[0370] Example 10: Synthesis of ADC3

[0371] Based on the amount of antibody, 2.7 molar equivalents of TCEP were added to the hRS9 antibody, and the pH was adjusted to 7 with 1 M Tris base. The solution was incubated at 25°C for 2 hours to reduce the concentration. TCEP was removed by ultrafiltration using 10 mM succinic acid. The thiol concentration of the antibody was determined by measuring the absorbance at 412 nm of the reaction product of the thiol group with DTNB (5,5'-dithiobis(2-nitrobenzoic acid), Aldrich).

[0372] During the coupling reaction, 6 molar equivalents of MC-GGFG-Dxd (Deruxtecan, MCE, Cat.# HY-13631E / CS-0045125) were added based on the amount of antibody. After stirring at 25°C for 2 hours, 0.1M acetylcysteine ​​was added to a final concentration of 2 mM and the reaction was terminated by stirring for 15 minutes. The reaction mixture was filtered through a 0.22 μm filter and eluted with 10 mM succinic acid through Sephadex G-25 resin.

[0373] The protein concentration of ADC3 was determined to be 2.83 mg / mL, and the DAR, i.e., p, was 4.6 as detected by RP-HPLC.

[0374] Example 11: Synthesis of ADC4

[0375] Based on the amount of antibody, 4.5 molar equivalents of TCEP were added to the hRS9 antibody, and the pH was adjusted to 8 with 1 M Tris base. The solution was incubated at 25°C for 2 hours to reduce the concentration. TCEP was removed by ultrafiltration using 10 mM succinic acid. The thiol concentration of the antibody was determined by measuring the absorbance at 412 nm of the reaction product of the thiol group with DTNB (5,5'-dithiobis(2-nitrobenzoic acid), Aldrich).

[0376] During the coupling reaction, 12 molar equivalents of CL2A-SN38 (see US2018161440A1 for details) were added based on the amount of antibody. After stirring at 25°C for 2 hours, 0.1M acetylcysteine ​​was added to a final concentration of 2 mM and the reaction was terminated by stirring for 15 minutes. The reaction mixture was filtered through a 0.22 μm filter and eluted with 10 mM succinic acid on Sephadex G-25 resin.

[0377] The protein concentration of ADC4 was determined to be 6.79 mg / mL, and the DAR, i.e., p, was approximately 8.3 as detected by RP-HPLC.

[0378] Example 12: In vitro biological activity of ADC1

[0379] We evaluated the growth inhibition of ADC1 using the Trop2-positive breast tumor cell lines MDA-MB-453, MDA-MB-468, and MX-1, as well as the Trop2-negative cell line HGC27 (purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences). Briefly, cells were grown to the logarithmic growth phase, trypsinized, detached, and suspended in 100 μL of complete culture medium. 4,000–8,000 cells were seeded in 96-well plates and cultured at 37°C for 3–5 hours or overnight. Then, 100 μL of culture medium containing varying concentrations of ADC1 was added. After 120 hours, the culture medium was removed from the culture dishes, and relative cell proliferation was analyzed using the Cell Counting Kit-8 (CCK-8, Dojindo Chemical). The results showed that ADC1 had a growth inhibitory effect on all Trop2-positive cells, with an EC50 of >15,000 ng / mL for the Trop2-negative HGC27 cell line.

[0380] Table 2

[0381] Example 13: Bystander Effect of ADC1 and ADC2

[0382] Trop2-positive MDA-MB-468 cells and Trop2-negative HGC27 cells were seeded into 6-well plates at a density of 3:1 (150,000 + 50,000). After culturing for 3-5 hours, ADC1 and ADC2, as well as the control drug ADC3, were added at final concentrations of 0.1 nM, 5 nM, and 50 nM, respectively. After further incubation at 37°C for 120 hours, the cells were trypsinized and washed once with PBS. The cells were counted and then labeled with FITC-labeled anti-TROP2 antibody at 4°C for 30 minutes to 1 hour. The ratio of different cell types was determined by flow cytometry, and the ratio of each cell type in the culture dish after different treatments was calculated, as shown in Figure 1. Bystander effects were observed for ADC1, ADC2, and ADC3, with ADC1 having a stronger bystander effect than ADC2 and the control drug ADC3.

[0383] Example 14: Pharmacokinetics in rats

[0384] Healthy adult Sprague Dawley rats, aged 6-8 weeks, were administered a tail vein bolus injection of ADC1 (5 mL / kg, 20 mg / kg) for approximately 1 minute ± 10 seconds. Blood was collected at 0.083, 1, 2, 8, 24, 48, 72, 96, 120, 144, and 168 hours after administration. Serum was separated by centrifugation within 30-120 minutes. Total antibody (Tab) and ADC concentrations in the blood samples were determined by conventional ELISA.

[0385] The total antibody detection method is briefly described as follows: Trop2-His was coated overnight at 4°C at a concentration of 0.75 μg / mL, and the plate was blocked with 5% skim milk powder at 37°C for 2 h. The standard curve and quality control point samples were incubated for 2 h. The detection range of the standard curve was 64 ng / mL-0.5 ng / mL, starting at 64 ng / mL, and the serial dilution was 2-fold. The quality control concentration points were set at 60 ng / mL, 6 ng / mL, and 0.6 ng / mL. The recovery rate of the quality control point needed to be between 80% and 120%. Subsequently, anti-human κ light chain peroxidase antibody produced in goat (manufacturer: Sigma, product number: A7164-1ML) was added at a dilution of 1:8000. The secondary antibody was incubated for 1 h, washed 8 times with PBST, and TMB was added for color development. The plate was terminated with 0.1 M sulfuric acid. The plate was read at OD450 on an enzyme-linked microplate reader. The blood sample concentration at different time points was calculated using the enzyme-linked microplate reader analysis software SoftMax Pro.

[0386] The ADC detection method is briefly described as follows: Trop2-His was coated overnight at 4°C at a concentration of 0.75 μg / mL, and blocked with 5% skim milk powder at 37°C for 2 hours. The standard curve and quality control point samples were added and incubated for 2 hours. The standard curve detection range was 64 ng / mL-0.5 ng / mL, starting at 64 ng / mL, and 2-fold serial dilution. The quality control concentration points were set at 60 ng / mL, 6 ng / mL, and 0.6 ng / mL. The quality control point recovery rate needed to be between 80% and 120%. Subsequently, a rabbit polyclonal antibody (obtained from GenScript Immunofluorescence) was added (diluted at 1:5000), incubated for 1 hour, and Fc fragment-specific peroxidase affinity pure goat anti-rabbit IgG (Jackson immunofluorescence) was added at a dilution of 1:8000. The plate was incubated for 1 hour and washed 8 times with PBST. 100 μL of single-component TMB colorimetric solution (Huzhou Yingchuang, TMB-S-001) was added for color development and terminated with 0.1 M sulfuric acid. The plate was read on an OD450 microplate reader. The concentrations of blood samples at different time points were calculated using the SoftMax Pro analysis software for the microplate reader. The curve of drug concentration in the blood was drawn using ELISA, as shown in Figure 2. The ADC1 concentration was basically consistent with the total antibody concentration, with extremely low shedding, and the plate was very stable in the blood.

[0387] Example 15: In vivo efficacy of ADC1 in Capan-1

[0388] This study evaluated the pharmacodynamics of ADC1 in a female BALB / c nude mouse model harboring subcutaneous xenografts of the human pancreatic cancer Capan-1 cell line. Ten mice were included in each group. The experimental design was as follows: ADC1 was the test drug, and ADC3 and ADC4 were the control drugs.

[0389] Table 3. Administration routes, doses, and regimens in the Capan-1 subcutaneous human pancreatic cancer xenograft model Note: 1. The day of grouping is defined as Day 0, and drug administration starts from Day 0.

[0390] All drugs were diluted with PBS to a given concentration before administration. The experimental animals were 7-9 week old BALB / c nude female mice (Beijing Ankai Yibo Biotechnology Co., Ltd.).

[0391] Capan-1 cells were cultured in IMDM medium containing 20% ​​fetal bovine serum. Capan-1 cells were collected during the exponential growth phase and resuspended in PBS to a suitable concentration for subcutaneous tumor inoculation in nude mice. 5×106 Capan-1 cells were subcutaneously inoculated at the right anterior scapula of the experimental mice. The cells were resuspended in PBS mixed with Matrigel at a ratio of 1:1 (0.1 mL / mouse). Tumor growth was observed regularly. When the tumors grew to an average volume of 159.77 mm3, they were randomly divided into groups and dosed according to tumor size and mouse weight. The day of grouping was set as day 0, and dosing began on day 0.

[0392] After tumor inoculation, routine monitoring included tumor growth and the effects of treatment on the animals' normal behavior. Specifically, the animals' activity, food and water intake, weight gain or loss (weight was measured twice weekly), and eye, coat, and other abnormalities were recorded in the raw data. Tumor volume was calculated using the formula: tumor volume (mm³) = 1 / 2 × (a × b²) (where a represents the major diameter and b represents the minor diameter). Data were collected using StudyDirector™ (version 3.1.399.19, Studylog System, Inc., San Francisco, CA, USA).

[0393] Table 4. Efficacy analysis of each group in Capan-1 xenograft model Note: a. Data are expressed as "mean ± standard error";

[0394] b. TGI% = [1-(Ti-T0) / (Ci-C0)] × 100, where T0 and C0 are the average tumor volumes of the drug-treated group and the vehicle control group on the day of grouping (day 0), and Ti and Ci are the average tumor volumes of the drug-treated group and the vehicle control group on day 25, respectively;

[0395] c. *P<0.05, **P<0.01 and ***P<0.001 compared with the tumor volume in the vehicle control group.

[0396] As shown in Figure 3, at a dose of 5 mg / kg, ADC1 was significantly more potent than the control drug ADC4 and ADC3. At a concentration of 5 mg / kg, ADC4 had essentially no inhibitory effect on tumor growth in this model. After a single dose of ADC1, the TGI of the tumor was as high as 114% on day 25.

[0397] Example 16: ADC1 and ADC3 in human triple-negative breast cancer MX-1 cell line subcutaneously xenografted female

[0398] Pharmacodynamics in BALB / c nude mouse model

[0399] Test drug: ADC1 (2.5 mg / kg and 5 mg / kg)

[0400] ADC3 (2.5 mg / kg and 5 mg / kg)

[0401] Preparation method: dilute with PBS

[0402] Experimental animals: BALB / c nude, 6 animals per group (Beijing Ankai Yibo Biotechnology Co., Ltd.).

[0403] Experimental Methods: MX-1 cells were cultured in RPM1640 medium supplemented with 10% fetal bovine serum. MX-1 cells were collected during the exponential growth phase and resuspended in PBS to a suitable concentration for subcutaneous tumor inoculation in nude mice. Mice were subcutaneously inoculated with 5×106 MX-1 cells on the right side of their backs. The cells were resuspended in PBS (0.1 mL / mouse). Tumor growth was regularly observed. When tumors grew to an average volume of 149.03 mm3, mice were randomly divided into groups for dosing based on tumor size and body weight. Grouping was designated Day 0, and dosing began on Day 0.

[0404] After tumor inoculation, routine monitoring included tumor growth and the effects of treatment on the animals' normal behavior.

[0405] Experimental Results: Tumor volume changes are shown in Table 5 and Figure 4. The results show that ADC1 and ADC3 had similar efficacy at a single dose, with TGIs of 109.48% and 108.84%, respectively, at a 5 mg / kg dose. No significant abnormalities or weight loss were observed in any of the mice groups during the experiment.

[0406] Table 5 Note: a. Data are expressed as "mean ± standard error";

[0407] b. TGI% = [1-(Ti-T0) / (Ci-C0)] × 100, where T0 and C0 are the average tumor volumes of the drug-treated group and the vehicle control group on the day of grouping (day 0), and Ti and Ci are the average tumor volumes of the drug-treated group and the vehicle control group on day 21, respectively;

[0408] c. *P<0.05, **P<0.01 and ***P<0.001 compared with the tumor volume in the vehicle control group.

[0409] Example 17: Preclinical Pharmacokinetic Study of ADC1

[0410] This study used 18 Cynomolgus monkeys were randomly divided into three groups, half male and half female. Groups 1-3 received ADC1 at doses of 1 mg / kg, 3 mg / kg, and 10 mg / kg, respectively, via a single intravenous infusion over 30 minutes. Blood samples were collected 43 days after administration (1008 hours). Exatecan concentrations in plasma samples were analyzed by LC-MS / MS, and ADC1 and total antibodies in serum samples were analyzed by ELISA.

[0411] After a single intravenous injection of 1 mg / kg, 3 mg / kg, and 10 mg / kg (dose ratio of 1:3:10) of ADC1 in cynomolgus monkeys, the Cmax and AUC of total antibody in the serum of male animals were (0-t) The ratios of ADC in male animals were 1:2:9 and 1:3:11 respectively, and those in female animals were 1:3:9 and 1:4:13 respectively; the C max and AUC (0-t) The ratios of Exatecan in male and female animals were 1:2:8 and 1:2:11, respectively, and those in female animals were 1:2:10 and 1:3:13, respectively. max and AUC (0-t) The intergroup ratios of male and female AUC were 1:2:6 and 1:70:768 respectively. (0-t) The ratio ranged from 0.85 to 1.48. The results showed that after a single intravenous injection of 1 mg / kg, 3 mg / kg, and 10 mg / kg of ADC1 in cynomolgus monkeys, total antibody and ADC systemic exposure in serum increased with increasing dose, with the increase ratio being roughly consistent with the dose increase ratio.

[0412] The pharmacokinetic study results of ADC1 administered as a single intravenous infusion to cynomolgus monkeys showed that there was no significant gender difference in exposure between males and females in each dose group.

[0413] Example 18: Human clinical study of ADC1

[0414] This study is a multicenter, open-label Phase I clinical study of the safety, tolerability, pharmacokinetic (PK) characteristics and preliminary efficacy of injectable ADC1 in patients with advanced solid tumors.

[0415] The trial patients are patients with advanced or metastatic epithelial solid tumors confirmed by pathological histology or cytology, who have failed standard treatment or have no standard treatment, are intolerant to standard treatment or refuse standard treatment, including but not limited to: urothelial carcinoma, triple-negative breast cancer, gastric adenocarcinoma, esophageal cancer, hepatocellular carcinoma, non-small cell lung cancer, small cell lung cancer, epithelial ovarian cancer, cervical cancer, follicular thyroid carcinoma, glioblastoma multiforme, endometrial cancer, prostate cancer, head and neck squamous cell carcinoma, and renal clear cell carcinoma.

[0416] The clinical study of ADC1 is divided into two phases. The first phase (dose-escalation tolerance study) plans to use accelerated titration and a "3+3" dose-escalation rule to explore the safety and tolerability of the study drug. Seven dose groups will be set up, namely A (0.8mg / kg), B (1.2mg / kg), C (2.4mg / kg), D (2.7mg / kg), E (3.0mg / kg), F (3.3mg / kg), and G (3.6mg / kg). The 0.8mg / kg group will use an accelerated titration method for dose escalation, and the remaining dose groups will use the standard "3+3" rule for dose escalation studies. The second phase (expansion cohort study) will select appropriate doses and tumor types for expansion studies based on the preliminary safety and efficacy results of the previous phase to further explore the safety and clinical efficacy of ADC1 and provide a basis for subsequent clinical studies.

[0417] The dose escalation rules of the "3+3" rule are as follows:

[0418] The first dose group adopts an accelerated titration dose escalation scheme, that is, the cohort size = 1. First, one subject in Group A is included. If no ≥ grade 2 toxicity related to the study drug is observed within 21 days, the subject can directly enter the next group for dose escalation. By analogy with this rule ("1+1"), the dose escalation is accelerated to the standard 3+3 group. If, during the accelerated escalation process, a ≥ grade 2 toxicity related to the study drug is observed in a certain group, the accelerated titration dose escalation is stopped, and two more subjects are required to be included in the dose group to switch to the standard "3+3" dose escalation (i.e., cohort size = 3) mode until the MTD or MAD is reached. Starting from Group B, the dose escalation mode is adjusted to the standard "3+3" rule. The escalation mode is shown in the following table:

[0419] When conducting tolerance exploration according to the "3+3" dose escalation rule, all subjects in the previous dose group must complete a 21-day DLT observation period before the next dose group cohort is enrolled. During the dose escalation study, the first dose of the previous and next subjects in each dose group must be at least 72 hours apart.

[0420] After the completion of the above-mentioned escalation study, after a comprehensive assessment of factors such as safety, it may be considered whether to add a higher dose level group to further explore tolerance; or, based on specific circumstances and discussions with researchers, a new dose may be selected between the dose at which the escalation is stopped and the previous lower dose group, and the test may be conducted according to the above rules to determine a more accurate maximum tolerated dose.

[0421] AEs were graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) version 5.0. DLT was defined as any AE of grade 3 or higher related to the trial drug occurring within 21 days of the first dose of the subject and meeting the following definitions:

[0422] Grade 5 toxicity.

[0423] Definition of hepatotoxicity DLT:

[0424] Grade 4: elevated AST (aspartate aminotransferase) or ALT (alanine aminotransferase);

[0425] Subjects with hepatocellular carcinoma or liver metastasis, if AST or ALT is ≤ 3 times ULN (upper limit of normal) at baseline, and AST or ALT increases to > 5 times ULN and persists for > 5 days during the DLT evaluation period;

[0426] Subjects with hepatocellular carcinoma or liver metastasis, if AST or ALT is greater than 3 times ULN at baseline, and AST or ALT increases to greater than 8 times ULN and persists for greater than 5 days during the DLT evaluation period;

[0427] In subjects without liver metastasis, AST or ALT elevation >5 times ULN persists for >5 days;

[0428] AST or ALT > 5 times the upper limit of normal range (ULN), accompanied by ≥ grade 2 elevated blood bilirubin.

[0429] Definition of hematologic toxicity DLT:

[0430] Grade 4 neutropenia lasting for >7 days;

[0431] ≥Grade 3 neutropenia with fever (single body temperature > 38.3 or continuous body temperature ≥ 38°C for more than 1 hour);

[0432] Grade 4 anemia;

[0433] Grade 4 thrombocytopenia;

[0434] ≥Grade 3 thrombocytopenia lasting for >7 days;

[0435] ≥Grade 3 thrombocytopenia with bleeding.

[0436] Definition of non-hepatotoxicity and non-hematologic toxicity DLTs:

[0437] Other grade ≥3 non-hepatotoxicity and non-hematological toxicity.

[0438] The following TEAEs were not considered DLTs:

[0439] Grade 3 nausea, vomiting, diarrhea, anorexia, and fatigue are controlled with standard supportive care and resolve to ≤ Grade 2 within 3 days;

[0440] Grade 3 skin and mucosal toxicity was controlled by standard treatment and resolved to ≤ Grade 2 within 7 days

[0441] Laboratory abnormalities without clinically relevant symptoms or signs, including elevated ALP, elevated uric acid, elevated blood amylase, elevated lipase, or lymphocytopenia to Grade 3 or 4, or Grade 1 hyponatremia at baseline upgraded to Grade 3 but lasting for less than 72 hours;

[0442] Grade 3 infusion reactions can be relieved within 6 hours after treatment.

[0443] Dosage regimen: Intravenous infusion, once every 2 weeks (Q2W) or once every 3 weeks (Q3W), administered on the first day of each cycle. The infusion should be started slowly. If vital signs are stable and no infusion reaction is found, the infusion rate can be gradually increased according to the following recommendations:

[0444] In the event of an infusion reaction or abnormal vital signs, the investigator may decide to slow, interrupt, or terminate the study drug infusion based on the specific clinical situation. It is recommended that the infusion be controlled for ≥3 hours for the first dose. If the initial dose is well tolerated, subsequent doses can be controlled within 1-2 hours. Closely monitor vital signs and for infusion reactions during and within 30 minutes after dosing. After the infusion, the infusion line should be slowly flushed with normal saline.

[0445] Tolerability and safety evaluation:

[0446] Tolerability evaluation indicators: dose-limiting toxicity (DLT) events and their incidence.

[0447] Safety evaluation indicators: vital signs, physical examination, laboratory tests (blood routine, blood biochemistry, cardiac function, thyroid function, coagulation test, urine routine, stool routine, pregnancy test), ECOG score, electrocardiogram, cardiac ultrasound, adverse events, etc. Assess the nature, frequency, severity, and time of occurrence of adverse events; evaluate changes in clinical laboratory test results before, during, and after treatment.

[0448] Pharmacokinetic evaluation:

[0449] Blood samples will be collected from subjects in all dose groups at defined time points during treatment (for the first six treatment cycles) to investigate pharmacokinetic characteristics. Serum concentrations of ADC1, total anti-Trop2 antibody, and Exatecan will be measured at the protocol-specified time points. Approximately 4.0 mL of blood will be collected at each time point. Unplanned PK sampling is recommended in the event of unexpected events (e.g., infusion reactions).

[0450] The following PK parameters (pharmacokinetics) were calculated using the actual dose, actual sampling time, and non-compartmental model: For single administration, the parameters included C max 、T max 、T 1 / 2 , CL, Vd, Ke, MRT, AUC (0-τ) , AUC (0-∞) ; For multiple doses, parameters include C max,ss 、C avg,ss 、C min,ss , AUC (0-τ),ss , AUC (0-∞),ss 、T max,ss 、T 1 / 2,ss 、C L 、V ss , Ke, MRT, accumulation index (Rac), volatility index DF.

[0451] Effectiveness evaluation indicators:

[0452] Anti-tumor efficacy indicators include: objective response rate (ORR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS) and overall survival (OS) based on the RECIST evaluation criteria and imaging evaluation using CT / MRI as the main evaluation method.

Claims

1. A method for treating solid tumors, characterized in that: The method comprises administering an effective amount of an antibody-drug conjugate to a patient with a solid tumor; wherein the effective amount of the antibody-drug conjugate is administered once every 1-8 weeks, with a dose of 0.1-30 mg / kg per administration, and the antibody-drug conjugate has a structure as shown in Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt or solvate thereof: in, Abu is an antibody or antigen-binding unit thereof that binds to TROP2; D is the drug; M is Wherein * connects to Abu, ** connects to B, and R is selected from: -(CH2) r -、-(CHR m ) r -, C3-C8 carbocyclic group, -O-(CH2) r -, arylene, -(CH2) r -Arylene-, -Arylene-(CH2)r-, -(CH2) r -(C3-C8 carbocyclyl)-, -(C3-C8 carbocyclyl)-(CH2) r -, C3-C8 heterocyclic group, -(CH2) r -(C3-C8 heterocyclyl)-, -(C3-C8 heterocyclyl)-(CH2) r -、-(CH2) r C(O)NR m (CH2) r -、-(CH2CH2O) r -、-(CH2CH2O) r -CH2-, -(CH2) r C(O)NR m (CH2CH2O) r -、-(CH2) r C(O)NR m (CH2CH2O) r -CH2-, -(CH2CH2O) r C(O)NR m (CH2CH2O) r -、-(CH2CH2O) r C(O)NR m (CH2CH2O) r -CH2- and -(CH2CH2O) r C(O)NR m (CH2) r -; each R m R is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each r is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; B is Where * connects to M, ** connects to L, and *** connects to G; L is -(AA) i -(FF) f -, wherein each AA is independently an amino acid or a polypeptide, i is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20; each FF is independently Among them, each R F is independently C1-C6 alkyl, C1-C6 alkoxy, -NO2 or halogen; z is 0, 1, 2, 3 or 4, wherein * is connected to AA and ** is connected to D; f is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; G is Where n is 1-24; p is 1-10.

2. The method according to claim 1, wherein The B is Among them, * connects to M, ** connects to L, and *** connects to G.

3. The method according to claim 1 or 2, wherein: Each AA is independently selected from the following amino acid or peptide sequences: Val-Cit, Val-Lys, Phe-Lys, Lys-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp, Cit, Phe-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Leu-Ala-Leu, Ile-Ala-Leu, Val-Ala-Val, Ala-Leu-Ala-Leu, β-Ala-Leu-Ala-Leu, and Gly-Phe-Leu-Gly.

4. The method according to claim 3, wherein The AA is Val-Cit.

5. The method according to any one of claims 1 to 4, characterized in that i is 1.

6. The method according to any one of claims 1 to 5, wherein: FF is independent Where * connects AA and ** connects D.

7. The method according to claim 6, wherein FF is Where * connects AA and ** connects D.

8. The method according to any one of claims 1 to 7, wherein: f is 1.

9. The method according to claim 8, wherein L is Where * connects to B and ** connects to D.

10. A method for treating solid tumors, characterized in that: The method comprises administering an effective amount of an antibody drug conjugate to a patient with a solid tumor; wherein the effective amount of the antibody drug conjugate is administered once every 1-8 weeks, and the dose of each administration is 0.1-30 mg / kg, and the antibody drug conjugate has the structure shown in Formula I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10 or I-11, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein The formula I-1 is: The formula I-2 is: The formula I-3 is: The formula I-4 is: The formula I-5 is: The formula I-6 is: The formula I-7 is: The formula I-8 is: The formula I-9 is: The formula I-10 is: The formula I-11 is: in Abu is an antibody or antigen-binding unit thereof that binds to TROP2; R is selected from: -(CH2) r -、-(CHR m ) r -, C3-C8 carbocyclic group, -O-(CH2) r -, arylene, -(CH2) r -Arylene-, -Arylene-(CH2)r-, -(CH2) r -(C3-C8 carbocyclyl)-, -(C3-C8 carbocyclyl)-(CH2) r -, C3-C8 heterocyclic group, -(CH2) r -(C3-C8 heterocyclyl)-, -(C3-C8 heterocyclyl)-(CH2) r -、-(CH2) r C(O)NR m (CH2) r -、-(CH2CH2O) r -、-(CH2CH2O) r -CH2-, -(CH2) r C(O)NR m (CH2CH2O) r -、-(CH2) r C(O)NR m (CH2CH2O) r -CH2-, -(CH2CH2O) r C(O)NR m (CH2CH2O) r -、-(CH2CH2O) r C(O)NR m (CH2CH2O) r -CH2- and -(CH2CH2O) r C(O)NR m (CH2) r -; each R m R is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each r is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; D is the drug; n is an integer from 1 to 24; p is 1-10.

11. The method according to any one of claims 1 to 10, wherein: The R is -(CH2)r-, and the r is 1 or 5.

12. The method according to any one of claims 1 to 11, wherein: The drug is an anticancer drug, a cytotoxic drug, a cell differentiation factor, a stem cell trophic factor, a steroid drug, a drug for treating autoimmune diseases, an anti-inflammatory drug or a drug for treating infectious diseases; or the drug is an anticancer drug; or the drug is a microtubule inhibitor, a DNA damaging agent or a DNA topoisomerase inhibitor; or the microtubule inhibitor is selected from dolastatin, auristatins, and maytansines; or the drug is an auristatin selected from MMAE, MMAF or AF; or the drug The drug is a DNA damaging agent selected from calicheamicins, duocarmycins, and anthramycin derivatives PBD; or the drug is a DNA topoisomerase inhibitor or a salt thereof selected from irinotecan, irinotecan hydrochloride, isotecan derivatives, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, camptothecin derivatives SN-38, 22-hydroxyecliptin, topotecan, lertotecan, belotecan, isotecan, silyl homocamptothecin, 6 ,8-dibromo-2-methyl-3-[2-(D-xylopyranosylamino)phenyl]-4(3H)-quinazolinone, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-(2E)-2-acrylamide, 2-cyano-3-(3,4-dihydroxyphenyl)-N-(3-hydroxyphenylpropyl)-(E)-2-acrylamide, 12-β-D-glucopyranosyl-12,13-dihydro-2,10-dihydroxy-6-[[2-hydroxy-1-(hydroxymethyl) [3,4-c]carbazole-5,7(6H)-dione, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide dihydrochloride, N-[2-(dimethylamino)ethyl]-4-acridinecarboxamide; or the DNA topoisomerase inhibitor is camptothecin, 10-hydroxycamptothecin, topotecan, belotecan, irinotecan, 22-hydroxyecilimine or exitecan; or a pharmaceutically acceptable salt or solvate thereof.

13. The method according to any one of claims 1 to 11, characterized in that The drug is in X 1 and X 2 Each independently is: H， hydroxyl groups, C1-C6 alkyl, C1-C6 alkyl substituted by one or more hydroxyl, halogen, nitro or cyano groups, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 aminoalkoxy, halogen, Nitro, cyano group, Thiol, Alkylthio, amino, amino substituted by an amino protecting group, C1-C6 aminoalkyl optionally substituted by an amino protecting group or a C1-C6 alkyl group at the amino moiety, C1-C6 aminoalkylamino optionally substituted with an amino protecting group or a C1-C6 alkyl group at the amino moiety, C1-C6 alkyl group connected to a heterocyclic ring, said heterocyclic ring being optionally substituted with one or more C1-C6 alkyl groups, C1-C6 alkoxy groups, amino groups, halogen groups, nitro groups or cyano groups, C1-C6 alkylamino group connected to a heterocyclic ring, the heterocyclic ring is optionally substituted by C1-C6 alkyl, C1-C6 alkoxy, the amino group is optionally substituted by an amino protecting group, halogen, nitro, cyano or a protecting group, An amino-substituted heterocyclic group, which is optionally substituted by a protecting group or one or more C1-C6 alkyl groups at the nitrogen atom of the heterocyclic ring or the amino group, Heterocyclic amino group, which is optionally substituted by a protecting group or a C1-C6 alkyl group at the nitrogen atom of the heterocyclic ring portion or the amino portion, a carbamoyl group optionally substituted with a carbamoyl protecting group or a C1-C6 alkyl group, Morpholin-1-yl, or Piperidin-1-yl; X 3 is a C1-C6 alkyl group; X 4 H, -(CH2) q -CH3、-(CHR n ) q -CH3, C3-C8 carbocyclyl, -O-(CH2) q -CH3, arylene-CH3, -(CH2) q -Arylene-CH3, -Arylene-(CH2) q -CH3, -(CH2) q -(C3-C8 carbocyclyl)-CH3, -(C3-C8 carbocyclyl)-(CH2) q -CH3, C3-C8 heterocyclic group, -(CH2) q -(C3-C8 heterocyclyl)-CH3, -(C3-C8 heterocyclyl)-(CH2) q -CH3, -(CH2) q C(O)NR n (CH2) q -CH3, -(CH2CH2O) q -CH3, -(CH2CH2O) q -CH2-CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH3, -(CH2) q C(O)NR n (CH2CH2O) q -CH2-CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH3, -(CH2CH2O) q C(O)NR n (CH2CH2O) q -CH2-CH3, or -(CH2CH2O) q C(O)NR n (CH2) q -CH3; each R n is independently H, C1-C6 alkyl, C3-C8 carbocyclyl, phenyl or benzyl; and each q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; or, X 4 is H or C1-C6 alkyl; ** is the connection point; y is 0, 1, or 2; Y is O, S or CR 1 R 2 , where R 1 and R 2 Each independently is H or C1-C6 alkyl; s and t are each independently 0, 1, or 2, but not both 0; Or the drug is where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; or the C1-C6 alkyl is -CH3; or the halogen is F; ** is the point of attachment; Or the drug is where X 1 and X 2 Each is independently C1-C6 alkyl, halogen or -OH; or the C1-C6 alkyl is -CH3; or the halogen is F; ** is the point of attachment.

14. The method according to any one of claims 1 to 13, wherein: n is 4-12; or n is 4-8; or n is 4 or 8.

15. A method for treating solid tumors, characterized in that: The method comprises administering an effective amount of an antibody drug conjugate to a patient having a solid tumor; wherein the effective amount of the antibody drug conjugate is administered once every 1-8 weeks, with a dose of 0.1-30 mg / kg each time, and the antibody drug conjugate has a structure as shown in Formula I-12, I-13, I-14, I-15, I-16, I-17, I-18, I-19, I-20, I-21, I-22, I-23, I-24 or I-25, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the Formula I-12, I-13, I-14, I-15, I-16, I-17, I-18, I-19, I-20, I-21, I-22, I-23, I-24 or I-25 is: in Abu is an antibody or antigen-binding unit thereof that binds to TROP2; p is 1-10.

16. The method according to any one of claims 1 to 15, wherein: p is 2-8; or p is 4-8; or p is 6-8; or p is 7-8.

17. The method according to any one of claims 1 to 16, characterized in that Abu is the hRS9 antibody.

18. The method according to any one of claims 1 to 16, characterized in that Abu is an antibody or an antigen-binding unit thereof that binds to TROP2, wherein the antibody or the antigen-binding unit thereof comprises a heavy chain shown in SEQ ID NO: 1 and a light chain shown in SEQ ID NO:

2.

19. The method according to any one of claims 1 to 18, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 0.1-30 mg / kg.

20. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose ranging from about 0.8 mg / kg to about 3.6 mg / kg.

21. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 2.4 mg / kg to about 3.6 mg / kg.

22. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 2.7 mg / kg to about 3.3 mg / kg.

23. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 0.8 mg / kg, about 1.2 mg / kg, about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg or about 3.6 mg / kg.

24. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 2.4 mg / kg, about 2.7 mg / kg, about 3.0 mg / kg, about 3.3 mg / kg or about 3.6 mg / kg.

25. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being about 3.0 mg / kg.

26. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-3000 mg.

27. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-2000 mg.

28. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-1500 mg.

29. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-1000 mg.

30. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-800 mg.

31. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 1, 2, 3, 4, 5, 6 or 7 weeks, with each dose being 5-600 mg.

32. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 2, 3 or 4 weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

33. The method according to any one of claims 1 to 32, characterized in that The effective amount of the antibody drug conjugate is administered once every 2 weeks.

34. The method according to any one of claims 1 to 32, wherein: The effective amount of the antibody drug conjugate is administered once every 3 weeks.

35. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

36. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every three weeks, with each dose ranging from about 2.4 mg / kg to about 3.6 mg / kg.

37. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every 2 weeks, with each dose ranging from about 2.7 mg / kg to about 3.3 mg / kg.

38. The method according to any one of claims 1 to 19, characterized in that The effective amount of the antibody drug conjugate is administered once every three weeks, with each dose ranging from about 2.7 mg / kg to about 3.3 mg / kg.

39. The method according to any one of claims 1 to 38, characterized in that The antibody drug conjugate is administered by injection; or by intravenous injection, subcutaneous injection or intraperitoneal injection; or by intravenous infusion.

40. The method according to claim 39, wherein The antibody drug conjugate is intravenously infused at a rate of ≤1000 mg / hr; or, the antibody drug conjugate is first intravenously infused at a rate of ≤500 mg / hr, and each subsequent intravenous infusion rate is ≤1000 mg / hr; or, when the antibody drug conjugate is intravenously infused, the rate from 0 to 15 minutes is ≤200 mg / hr, and starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 50-200 mg / hr; or, when the antibody drug conjugate is first intravenously infused, the rate from 0 to 15 minutes is ≤50 mg / hr, and starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 50 mg / hr, and for each subsequent injection, the rate from 0 to 15 minutes is 100-200 mg / hr, and starting from the 15th minute, the rate can be increased every 15 to 30 minutes, with a gradual increase of 100-200 mg / hr.

41. The method according to any one of claims 1 to 40, characterized in that The solid tumor is a TROP2-positive solid tumor or an advanced solid tumor, and the solid tumor includes but is not limited to urothelial carcinoma, triple-negative breast cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, glioblastoma, medulloblastoma, breast cancer, head and neck cancer, kidney cancer, ovarian cancer, gastric cancer, Kaposi's sarcoma, lung cancer, cervical cancer, colorectal cancer, esophageal cancer, oral squamous cell carcinoma, prostate cancer, thyroid cancer, bladder cancer, glioma, hepatobiliary carcinoma, colorectal cancer, T cell lymphoma, colorectal cancer, prostate cancer, melanoma, and liver cancer.

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