Use of Anti-trop2 antibody-drug conjugate in combination with Anti-PD-l1 / CD47 bispecific antibody for treating tumors

Abstract

The present invention relates to a method of using or a use of an anti-TROP2 antibody-drug conjugate in combination with an anti-PD-L1 / CD47 bispecific antibody or an antigen-binding fragment for treating tumors.

Description

Use of anti-TROP2 antibody-drug conjugates and anti-PD-L1 / CD47 bispecific antibodies in combination therapy for tumors. Technical Field

[0001] This invention relates to the field of drug therapy, specifically to the use of anti-TROP2 antibody-drug conjugates and anti-PD-L1 / CD47 bispecific antibodies in combination for the treatment of tumors. Background Technology

[0002] Cancer is a common and fatal disease. Chemotherapy, as one of the standard treatments for advanced cancer, still plays an important role in clinical treatment. However, its poor selectivity and strong toxic side effects lead some patients to refuse it or be unable to tolerate it. Currently, there is a huge unmet clinical need for cancer treatment and prognosis. Exploring drug therapies or combination therapies targeting various mechanisms of action to benefit more patients is an urgent clinical problem to be solved. Summary of the Invention

[0003] TROP2 (trophoblast cell surface glycoprotein antigen 2) is overexpressed in various cancer cells. This invention provides a method or use of an anti-TROP2 antibody-drug conjugate and an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment for the treatment of tumors, comprising administering an effective amount of the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment to a patient in need. In one aspect, this invention provides the use of the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment in the preparation of a medicament for the treatment of tumors. In another aspect, this invention provides the use of the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment in combination for the treatment of tumors. In yet another aspect, this invention provides a pharmaceutical composition of an anti-TROP2 antibody-drug conjugate and an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment, and its use in the treatment of tumors. On one hand, this invention provides the use of an anti-TROP2 antibody-drug conjugate in the preparation of a medicament for the treatment of tumors in combination with an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment. On another hand, this invention provides the use of an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment in the preparation of a medicament for the treatment of tumors in combination with an anti-TROP2 antibody-drug conjugate. On yet another hand, this invention provides the use of an anti-TROP2 antibody-drug conjugate in the treatment of tumors in combination with an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment. On yet another hand, this invention provides the use of an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment in the treatment of tumors in combination with an anti-TROP2 antibody-drug conjugate.

[0004] In some embodiments, the anti-TROP2 antibody-drug conjugate has a structure as shown in Formula I-1 or Formula I-2, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0005] in

[0006] Abu is an anti-TROP2 antibody or an antigen-binding fragment;

[0007] D is a DNA topoisomerase I inhibitor or a salt thereof. For example, the DNA topoisomerase I inhibitor is selected from irinotecan, irinotecan hydrochloride, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, camptothecin derivative SN-38, 22-hydroxyeclipticine, topotecan, letopotecan, belotecone, homosilatecan, 6,8-dibromo-2-methyl-3-[2-(D-pyranoxylamino)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-acrylamide, N-[2-(dimethylamino)ethyl]-4-acrylamide, eczetidine or eczetidine derivatives;

[0008] n is an integer from 1 to 24, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24;

[0009] p is approximately 1-10, for example approximately 1, approximately 2, approximately 3, approximately 4, approximately 5, approximately 6, approximately 7, approximately 8, approximately 9, or approximately 10.

[0010] In some implementations, D is ixotecan or an ixotecan derivative.

[0011] In some implementations, D is

[0012] Where X 1 and X 2 Each is independently a C1-C6 alkyl, halogen, or -OH; ** represents a connection point.

[0013] In some embodiments, the C1-C6 alkyl group is -CH3.

[0014] In some implementations, the halogen is F.

[0015] In some implementation schemes, X 1 and X 2 Each can be independently -CH3, F, or -OH.

[0016] In some implementation schemes, X 1 and X 2 Each is -CH3.

[0017] In some implementation schemes, X 1 and X 2 Each can be F, Cl, Br, or I independently.

[0018] In some implementation schemes, X 1 and X 2 Each is F.

[0019] In some implementation schemes, X 1 and X 2 Each can be either F or -CH3.

[0020] In some implementation schemes, X 1 For -CH3 and X 2 It is F.

[0021] In some implementations, n is an integer from 4 to 12. In some implementations, n is an integer from 4 to 8. In some implementations, n is 4. In some implementations, n is 8.

[0022] In some embodiments, p is about 2-8. In some embodiments, p is about 4-8. In some embodiments, p is about 4-5. In some embodiments, p is about 6-8. In some embodiments, p is about 7-8. In some embodiments, p is about 4-6. In some embodiments, p is about 5-7. In some embodiments, p is about 5.5-6.5. In some embodiments, p is about 4. In some embodiments, p is about 6.

[0023] In some embodiments, the anti-TROP2 antibody-drug conjugate has a structure as shown in formula I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12, I-13, I-14, I-15, I-16, I-17, or I-18, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof.

[0024] in,

[0025] Abu is an anti-TROP2 antibody or an antigen-binding fragment;

[0026] n is an integer from 1 to 24, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24;

[0027] p is approximately 1-10, for example approximately 1, approximately 2, approximately 3, approximately 4, approximately 5, approximately 6, approximately 7, approximately 8, approximately 9, or approximately 10.

[0028] In some implementations, n is an integer from 4 to 12. In some implementations, n is an integer from 4 to 8. In some implementations, n is 4. In some implementations, n is 8.

[0029] In some embodiments, p is about 2-8. In some embodiments, p is about 4-8. In some embodiments, p is about 4-5. In some embodiments, p is about 6-8. In some embodiments, p is about 7-8. In some embodiments, p is about 4-6. In some embodiments, p is about 5-7. In some embodiments, p is about 5.5-6.5. In some embodiments, p is about 4. In some embodiments, p is about 6.

[0030] In some implementations, Abu is an anti-TROP2 antibody.

[0031] In some implementations, the anti-TROP2 antibody can specifically target the Trop2 protein.

[0032] In some implementations, the anti-TROP2 antibody is a fully human monoclonal antibody.

[0033] In some implementations, the anti-TROP2 antibody is a humanized monoclonal antibody.

[0034] In some embodiments, the anti-TROP2 antibody is an antibody disclosed in the following patent documents: WO2021147993A1 (e.g., PD3), CN101264325B (e.g., RS7, hRS7), CN105849126B (e.g., hTINA1-H1L1, hTINA1-H2L1, hTINA1-H2L2, hTINA1-H3L3), CN110903395A (e.g., M1, M2, M3), CN1 13896796A (e.g., 4D3, 7F11), US20130089872 (e.g., K5-70, K5-107, K5-116-2-1, T6-16, T5-86), US5840854A (e.g., BR110), US20130122020 (e.g., 3E9, 6G11, 7E6, 15E2, 18B1), US20120237518 (e.g., 77220, KM4097, KM4590).

[0035] In some embodiments, the anti-TROP2 antibody is 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).

[0036] In some embodiments, the anti-TROP2 antibody is anti-TROP2 antibodies 162-25.3 and 162-46.2 disclosed by Lipinski et al. (1981, Proc. Natl. Acad. Sci. USA, 78: 5147-50) or Pr1E11 anti-TROP2 antibody disclosed by Ikeda et al. (2015, Biochem Biophys Res Comm 458: 877-82), which recognize a unique epitope on TROP2.

[0037] In some embodiments, the anti-TROP2 antibody comprises a heavy chain as shown in SEQ ID NO:19 and a light chain as shown in SEQ ID NO:20.

[0038] In some implementations, the anti-TROP2 antibody is antibody hRS9.

[0039] In some embodiments, the anti-TROP2 antibody-drug conjugate is ADC1 or its stereoisomer or a pharmaceutically acceptable salt or solvate thereof, wherein ADC1 has the structure shown in Formula II:

[0040] Where p is approximately 4-8.

[0041] In some embodiments, p is about 4-6. In some embodiments, p is about 5-7. In some embodiments, p is about 5.5-6.5. In some embodiments, p is about 4, about 5, about 6, about 7, or about 8. In some embodiments, p is about 6.

[0042] In some embodiments, the anti-TROP2 antibody-drug conjugate is ADC2 or its stereoisomer or a pharmaceutically acceptable salt or solvate thereof, wherein ADC2 has the structure shown in Formula III:

[0043] Where p is approximately 4-8.

[0044] In some embodiments, p is about 4-6. In some embodiments, p is about 5-7. In some embodiments, p is about 5.5-6.5. In some embodiments, p is about 4, about 5, about 6, about 7, or about 8. In some embodiments, p is about 6.

[0045] In some embodiments, the anti-TROP2 antibody-drug conjugate is the anti-TROP2 antibody-drug conjugate disclosed in WO2022 / 253284.

[0046] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment includes a variable region a that specifically binds to PD-L1; wherein the variable region a comprises one or more amino acid sequences from (a) to (f):

[0047] (a) VHa CDR1, which contains the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:1;

[0048] (b) VHa CDR2, which contains the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:2;

[0049] (c) VHa CDR3, which contains the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:3;

[0050] (d)VLa CDR1, which contains the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:4;

[0051] (e)VLa CDR2, which contains the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:5;

[0052] (f) VLa CDR3, which contains the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:6.

[0053] In some embodiments, the variable region a comprises a heavy chain variable region a (VHa), which comprises VHa CDR1, VHa CDR2, and VHa CDR3; wherein VHa CDR1 comprises the amino acid sequence shown in SEQ ID NO:1; VHa CDR2 comprises the amino acid sequence shown in SEQ ID NO:2; and VHa CDR3 comprises the amino acid sequence shown in SEQ ID NO:3.

[0054] In some embodiments, the variable region a comprises a light chain variable region a (VLa), which comprises VLa CDR1, VLa CDR2, and VLa CDR3; wherein VLa CDR1 comprises the amino acid sequence shown in SEQ ID NO:4; VLa CDR2 comprises the amino acid sequence shown in SEQ ID NO:5; and VLa CDR3 comprises the amino acid sequence shown in SEQ ID NO:6.

[0055] In some embodiments, the variable region a comprises VHa and VLa; VHa comprises VHa CDR1, VHa CDR2, and VHa CDR3; VLa comprises VLa CDR1, VLa CDR2, and VLa CDR3; wherein VHa CDR1 comprises the amino acid sequence shown in SEQ ID NO:1; VHa CDR2 comprises the amino acid sequence shown in SEQ ID NO:2; VHa CDR3 comprises the amino acid sequence shown in SEQ ID NO:3; VLa CDR1 comprises the amino acid sequence shown in SEQ ID NO:4; VLa CDR2 comprises the amino acid sequence shown in SEQ ID NO:5; and VLa CDR3 comprises the amino acid sequence shown in SEQ ID NO:6.

[0056] In some embodiments, the variable region a comprises VHa and VLa; VHa comprises an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:10; VLa comprises an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:12.

[0057] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment further comprises a light chain constant region a (CLa) and a heavy chain constant region a (CHa).

[0058] In some embodiments, the CHa comprises an amino acid sequence as shown in SEQ ID NO:13, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:13, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:13.

[0059] In some embodiments, the CLa comprises an amino acid sequence as shown in SEQ ID NO:14, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:14, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:14.

[0060] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody comprises a heavy chain a and a light chain a; the heavy chain a and the light chain a pair to form a PD-L1 antigen binding site.

[0061] In some embodiments, the heavy chain a comprises an amino acid sequence as shown in SEQ ID NO:16, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:16, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:16.

[0062] In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤10 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤1 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤0.5 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤0.2 nM.

[0063] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment comprises a variable region b that specifically binds to CD47; wherein the variable region b comprises one or more amino acid sequences in (g)-(l):

[0064] (g)VHb CDR1, which contains the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:7;

[0065] (h)VHb CDR2, which contains the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:8;

[0066] (i) VHb CDR3, which contains the amino acid sequence shown in SEQ ID NO:9, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:9;

[0067] (j)VLb CDR1, which contains the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:4;

[0068] (k)VLb CDR2, which contains the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:5;

[0069] (l)VLb CDR3, which contains the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:6.

[0070] In some embodiments, the variable region b comprises a heavy chain variable region b (VHb), which includes VHb CDR1, VHb CDR2, and VHb CDR3; wherein VHb CDR1 contains the amino acid sequence shown in SEQ ID NO:7; VHb CDR2 contains the amino acid sequence shown in SEQ ID NO:8; and VHb CDR3 contains the amino acid sequence shown in SEQ ID NO:9.

[0071] In some embodiments, the variable region b comprises a light chain variable region b (VLb), which includes VLb CDR1, VLb CDR2, and VLb CDR3; wherein VLb CDR1 contains the amino acid sequence shown in SEQ ID NO:4; VLb CDR2 contains the amino acid sequence shown in SEQ ID NO:5; and VLb CDR3 contains the amino acid sequence shown in SEQ ID NO:6.

[0072] In some embodiments, the variable region b comprises VHb and VLb; the VHb comprises VHb CDR1, VHb CDR2, and VHb CDR3; the VLb comprises VLb CDR1, VLb CDR2, and VLb CDR3; wherein, VHb CDR1 comprises the amino acid sequence shown in SEQ ID NO:7; VHb CDR2 comprises the amino acid sequence shown in SEQ ID NO:8; VHb CDR3 comprises the amino acid sequence shown in SEQ ID NO:9; VLb CDR1 comprises the amino acid sequence shown in SEQ ID NO:4; VLb CDR2 comprises the amino acid sequence shown in SEQ ID NO:5; and VLb CDR3 comprises the amino acid sequence shown in SEQ ID NO:6.

[0073] In some embodiments, the variable region b comprises VHb and VLb; the VHb comprises an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:11, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:11; the VLb comprises an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:12.

[0074] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment further comprises a light chain constant region b (CLb) and a heavy chain constant region b (CHb).

[0075] In some embodiments, the CHb comprises an amino acid sequence as shown in SEQ ID NO:15, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:15, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:15.

[0076] In some embodiments, the CLb comprises an amino acid sequence as shown in SEQ ID NO:14, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:14, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:14.

[0077] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody comprises a heavy chain b and a light chain b; the heavy chain b and the light chain b pair to form a CD47 antigen binding site.

[0078] In some embodiments, the heavy chain b comprises an amino acid sequence as shown in SEQ ID NO:17, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:17.

[0079] In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with CD47 is ≤10 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with CD47 is ≤5 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with CD47 is ≤4 nM.

[0080] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment does not cause erythrocyte agglutination.

[0081] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment includes a variable region a that specifically binds to PD-L1 and a variable region b that specifically binds to CD47.

[0082] Wherein, the variable region a contains one or more amino acid sequences from (a) to (f):

[0083] (a) VHa CDR1, which contains the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:1;

[0084] (b) VHa CDR2, which contains the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:2;

[0085] (c) VHa CDR3, which contains the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:3;

[0086] (d)VLa CDR1, which contains the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:4;

[0087] (e)VLa CDR2, which contains the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:5;

[0088] (f) VLa CDR3, which contains the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:6;

[0089] The variable region b contains one or more amino acid sequences from (g)-(l):

[0090] (g)VHb CDR1, which contains the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:7;

[0091] (h)VHb CDR2, which contains the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:8;

[0092] (i) VHb CDR3, which contains the amino acid sequence shown in SEQ ID NO:9, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:9;

[0093] (j)VLb CDR1, which contains the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:4;

[0094] (k)VLb CDR2, which contains the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:5;

[0095] (l)VLb CDR3, which contains the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:6.

[0096] In some embodiments, the variable region a comprises a heavy chain variable region a (VHa), which includes VHa CDR1, VHa CDR2, and VHa CDR3; wherein VHa CDR1 includes the amino acid sequence shown in SEQ ID NO:1; VHa CDR2 includes the amino acid sequence shown in SEQ ID NO:2; and VHa CDR3 includes the amino acid sequence shown in SEQ ID NO:3; the variable region b comprises a heavy chain variable region b (VHb), which includes VHb CDR1, VHb CDR2, and VHb CDR3; wherein VHb CDR1 includes the amino acid sequence shown in SEQ ID NO:7; VHb CDR2 includes the amino acid sequence shown in SEQ ID NO:8; and VHb CDR3 includes the amino acid sequence shown in SEQ ID NO:9.

[0097] In some embodiments, the variable region a comprises a light chain variable region a (VLa), which includes VLa CDR1, VLa CDR2, and VLa CDR3; wherein VLa CDR1 includes the amino acid sequence shown in SEQ ID NO:4; VLa CDR2 includes the amino acid sequence shown in SEQ ID NO:5; and VLa CDR3 includes the amino acid sequence shown in SEQ ID NO:6; the variable region b comprises a light chain variable region b (VLb), which includes VLb CDR1, VLb CDR2, and VLb CDR3; wherein VLb CDR1 includes the amino acid sequence shown in SEQ ID NO:4; VLb CDR2 includes the amino acid sequence shown in SEQ ID NO:5; and VLb CDR3 includes the amino acid sequence shown in SEQ ID NO:6.

[0098] In some embodiments, the variable region a comprises VHa and VLa; the variable region b comprises VHb and VLb; the VHa comprises VHa CDR1, VHa CDR2, and VHa CDR3; the VLa comprises VLa CDR1, VLa CDR2, and VLa CDR3; wherein, VHa CDR1 comprises the amino acid sequence shown in SEQ ID NO:1; VHa CDR2 comprises the amino acid sequence shown in SEQ ID NO:2; VHa CDR3 comprises the amino acid sequence shown in SEQ ID NO:3; the VHb comprises VHb CDR1, VHb CDR2, and VHb CDR3; the VLb comprises VLb CDR1, VLb CDR2, and VLb CDR3; wherein, VHb CDR1 comprises the amino acid sequence shown in SEQ ID NO:7; VHb CDR2 comprises the amino acid sequence shown in SEQ ID NO:8; VHb CDR3 comprises the amino acid sequence shown in SEQ ID NO:9; VLa CDR1 and / or VLb CDR1 comprises the amino acid sequence shown in SEQ ID NO:9. The amino acid sequence shown in NO:4; VLa CDR2 and / or VLb CDR2 contain the amino acid sequence shown in SEQ ID NO:5; VLa CDR3 and / or VLb CDR3 contain the amino acid sequence shown in SEQ ID NO:6.

[0099] In some embodiments, the VLa CDR1 contains the same amino acid sequence as the VLb CDR1.

[0100] In some embodiments, the VLa CDR2 contains the same amino acid sequence as the VLb CDR2.

[0101] In some embodiments, the VLa CDR3 contains the same amino acid sequence as the VLb CDR3.

[0102] In some embodiments, the VLa CDR1 and the VLb CDR1 contain the same amino acid sequence, the VLa CDR2 and the VLb CDR2 contain the same amino acid sequence, and the VLa CDR3 and the VLb CDR3 contain the same amino acid sequence.

[0103] In some embodiments, the variable region a comprises a heavy chain variable region VHa and a light chain variable region VLa; wherein, VHa comprises an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:10; and / or VLa comprises an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:12.

[0104] In some embodiments, the variable region b comprises a heavy chain variable region VHb and a light chain variable region VLb; wherein, the VHb comprises an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:11; and / or the VLb comprises an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:12.

[0105] In some embodiments, the VHa comprises an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:10; the VHb comprises an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:11, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:11.

[0106] In some embodiments, the VLa or VLb comprises an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:12.

[0107] In some embodiments, the variable region a comprises VHa and VLa; the variable region b comprises VHb and VLb; VHa comprises the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:10; VHb comprises the amino acid sequence shown in SEQ ID NO:11, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:11, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:11; VLa and / or VLb comprises the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:12.

[0108] In some implementations, the VLa and the VLb contain the same amino acid sequence.

[0109] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment further comprises a light chain constant region a (CLa), a heavy chain constant region a (CHa), a light chain constant region b (CLb), and a heavy chain constant region b (CHb).

[0110] In some embodiments, VHa is connected to CHa to form heavy chain a, and VHb is connected to CHb to form heavy chain b. In some embodiments, VLa is connected to CLa to form light chain a, and VLb is connected to CLa to form light chain b.

[0111] In some embodiments, CHa is the IgG1 subtype. In some embodiments, CHb is the IgG1 subtype.

[0112] In some embodiments, CHa and / or CHb comprise one or more of the following amino acid mutations of IgG1: Y349C, S354C, T366W, T366S, L368A, and Y407V, wherein the amino acid position is Eu numbered.

[0113] In some embodiments, CHa and / or CHb comprise the IgG1 subtype with the following amino acid mutation: N297A, wherein the amino acid position is Eu numbered.

[0114] In some embodiments, one of the heavy chain constant regions of CHa and CHb contains one or more of the following amino acid mutations: N297A, Y349C, T366S, L368A, and Y407V.

[0115] In some embodiments, another heavy chain constant region in CHa and CHb contains one or more amino acid mutations, namely N297A, S354C, and T366W.

[0116] In some embodiments, CHa comprises an amino acid sequence as shown in SEQ ID NO:13, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:13, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:13; CHb comprises an amino acid sequence as shown in SEQ ID NO:15, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:15, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:15.

[0117] In some embodiments, CHa comprises an amino acid sequence as shown in SEQ ID NO:13, and CHb comprises an amino acid sequence as shown in SEQ ID NO:15.

[0118] In some embodiments, the CLa comprises an amino acid sequence as shown in SEQ ID NO:14, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:14, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:14; the CLb comprises an amino acid sequence as shown in SEQ ID NO:14, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:14, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:14.

[0119] In some embodiments, heavy chain a comprises the amino acid sequence shown in SEQ ID NO:16, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:16, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:16; heavy chain b comprises the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:17. In some embodiments, light chain a and light chain b comprise the same amino acid sequence. In some embodiments, both light chain a and light chain b comprise the amino acid sequence shown in SEQ ID NO:18, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:18, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:18.

[0120] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody is antibody BsAb-71-N297A, which comprises heavy chain a, heavy chain b, light chain a, and light chain b. Heavy chain a comprises the amino acid sequence shown in SEQ ID NO:16, heavy chain b comprises the amino acid sequence shown in SEQ ID NO:17, and light chains a and b comprise the amino acid sequence shown in SEQ ID NO:18.

[0121] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody is the anti-PD-L1 / CD47 bispecific antibody disclosed in CN115702931A.

[0122] Table 1. Amino acid sequence of anti-PD-L1 / CD47 bispecific antibody

[0123] In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤10 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤1 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤0.5 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with PD-L1 is ≤0.2 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with CD47 is ≤10 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment with CD47 is ≤5 nM. In some embodiments, the affinity index (KD) of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment for CD47 is ≤4 nM. In some embodiments, the affinity of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment for PD-L1 is greater than its affinity for CD47.

[0124] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment has one or more of the following properties:

[0125] (a) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody can bind to both PD-L1 and CD47 simultaneously and maintain the affinity constant of the parent antibody, thereby blocking the PD1 / PD-L1 signaling pathway and blocking the SIRPα / CD47 signaling pathway.

[0126] (b) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody can bind to PD-L1 with ultra-high affinity (e.g., KD = 0.174 nM) and to CD47 with high affinity (e.g., KD = 3.78 nM); the specific binding of the anti-PD-L1 / CD47 bispecific antibody to PD-L1 on tumor cells promotes selective binding of the anti-PD-L1 / CD47 bispecific antibody to tumor cells, avoids binding to CD47 expressed in many normal tissues, and reduces side effects; the effective dose range of the anti-PD-L1 / CD47 bispecific antibody can be significantly expanded when the affinity for CD47 is much lower than that for PD-L1.

[0127] (c) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody has a stable four-chain antibody structure and amino acid residues that facilitate proper coupling or pairing between the chains. In some embodiments, the Fc domains of the anti-PD-L1 / CD47 bispecific antibodies respectively contain Y349C and S354C or S354C and Y349C respectively; the Fc domains of the PD-L1 / CD47 bispecific antibodies respectively contain protrusions (“knobs”) or holes (“holes”), and the protrusions or holes in the Fc domain of the first polypeptide chain can be placed in the holes or protrusions in the Fc domain of the third polypeptide chain, thereby forming a stable “knob-in-hole” association between the first polypeptide chain and the third polypeptide chain; wherein, the first polypeptide chain contains the structure VHa-CHa, and the first polypeptide chain and the light chain can specifically recognize and bind PD-L1; the third polypeptide chain contains the structure VHb-CHb, and the third polypeptide chain and the light chain can specifically recognize and bind CD47.

[0128] (d) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody has the advantages of being easy to express stably in cultured cells in vitro, having good thermal stability, high antibody yield and purity, and not requiring complex production processes.

[0129] (e) The anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment binds to cells expressing human PD-L1, in some embodiments, for example, at an EC50 concentration of less than or equal to about 2 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM. In some embodiments, the binding is measured by flow cytometry (e.g., FACS). In some embodiments, the cells expressing human PD-L1 are CHO-S cells expressing human PD-L1.

[0130] (f) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment blocks the relevant PD-L1 / PD-1 activity, for example, with an EC50 of less than or equal to about 0.3 μg / ml, 0.2 μg / ml, or 0.1 μg / ml, or with an EC50 of about 0.1-3 μg / ml, 0.1-0.4 μg / ml, or 0.1-0.3 μg / ml. In some embodiments, the relevant PD-L1 / PD-1 activity refers to the activation of inhibitory signals in the intracellular region of PD1 after PD-L1 binds to PD-1. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment inhibits the binding of PD-L1 to PD-1 in an MOA assay at an EC50 of less than or equal to, about less than or equal to, about 0.3 μg / ml, 0.2 μg / ml, or 0.1 μg / ml, or about 0.1-3 μg / ml, 0.1-0.4 μg / ml, or 0.1-0.3 μg / ml. In some embodiments, the cells are CHO cells overexpressing human PD-L1.

[0131] (g) The anti-PD-L1 / CD47 bispecific antibody does not cause significant cell agglutination; for example, the anti-PD-L1 / CD47 bispecific antibody does not cause significant erythrocyte hemagglutination. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody is considered not to cause significant agglutination if the agglutination level in the presence of the anti-PD-L1 / CD47 bispecific antibody decreases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% compared to the agglutination level in the presence of the anti-CD47 positive control antibody Hu5F9-G4 (Magrolimab). Preferably, the anti-PD-L1 / CD47 bispecific antibody does not cause significant cell agglutination when the antibody concentration is between 400 pM and 800 nM.

[0132] (h) The anti-PD-L1 / CD47 bispecific antibody does not bind to human erythrocytes. In one embodiment, if the erythrocyte binding level of the anti-PD-L1 / CD47 bispecific antibody decreases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% compared to the erythrocyte binding level in the presence of the anti-CD47 positive control antibody Hu5F9-G4, then the anti-PD-L1 / CD47 bispecific antibody does not significantly bind to human erythrocytes. Preferably, the anti-PD-L1 / CD47 bispecific antibody does not significantly bind to human erythrocytes when the antibody concentration is between 200 pM and 100 nM.

[0133] (i) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody is a blocking antibody that blocks the binding of CD47 to SIRPα. In some embodiments, the ability of macrophages to phagocytose tumor cells is increased by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% in the presence of the anti-PD-L1 / CD47 bispecific antibody.

[0134] (j) In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is capable of inducing antibody-dependent cell-mediated cytotoxicity (ADCC). In one embodiment, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is not capable of inducing antibody-dependent cell-mediated cytotoxicity (ADCC).

[0135] (k) The anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment synergistically inhibits one or more activities of PD-L1 and CD47.

[0136] In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 0.1-10 mg / kg. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1-5 mg / kg. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1.5-3 mg / kg. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 2-3 mg / kg. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 0.1 mg / kg, about 0.8 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 1.5 mg / kg, about 2.1 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 4.0 mg / kg, about 4.8 mg / kg, about 5.0 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) of any two of these values, or any value thereof. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered as a single dose. In some embodiments, it is administered every 1-8 weeks. In some embodiments, it is administered every 1-4 weeks. In some implementations, the drug is administered once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some implementations, the drug is administered once weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, or every 7 weeks. In some implementations, the drug is administered once every 2 weeks. In some implementations, the drug is administered once every 2 weeks after the initial 3 weeks.

[0137] In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 5-3000 mg per administration. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 10-500 mg per administration. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 5 mg, about 10 mg, about 50 mg, about 100 mg, about 126 mg, about 144 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 (including endpoints) of any two of these values, or any value thereof. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered as a single dose. In some embodiments, it is administered every 1-8 weeks. In some embodiments, it is administered every 1-4 weeks. In some embodiments, it is administered approximately every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, it is administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, or every 7 weeks. In some embodiments, it is administered every 2 weeks. In some embodiments, it is administered every 2 weeks after the initial 3-week dosing.

[0138] In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 0.1-10 mg / kg every 1-4 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1-5 mg / kg every 1-4 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1.5-3 mg / kg every 1-4 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 2-3 mg / kg every 1-4 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 1-4 weeks at approximately 0.1 mg / kg, approximately 0.8 mg / kg, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.5 mg / kg, approximately 2.1 mg / kg, approximately 2.4 mg / kg, approximately 2.7 mg / kg, approximately 3.0 mg / kg, approximately 3.3 mg / kg, approximately 3.6 mg / kg, approximately 4.0 mg / kg, approximately 4.8 mg / kg, approximately 5.0 mg / kg, approximately 6.0 mg / kg, approximately 7.2 mg / kg, approximately 8.4 mg / kg, or approximately 10 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0139] In some embodiments, approximately 5-3000 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 1-4 weeks. In some embodiments, approximately 10-500 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 1-4 weeks. In some embodiments, approximately 50-300 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 1-4 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered once every 1-4 weeks in amounts of about 5 mg, about 10 mg, about 50 mg, about 100 mg, about 126 mg, about 144 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 thereof.

[0140] In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 0.1-10 mg / kg every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1-5 mg / kg every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 1.5-3 mg / kg every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered at a dose of about 2-3 mg / kg every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered once every 2 weeks at approximately 0.1 mg / kg, approximately 0.8 mg / kg, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.5 mg / kg, approximately 2.1 mg / kg, approximately 2.4 mg / kg, approximately 2.7 mg / kg, approximately 3.0 mg / kg, approximately 3.3 mg / kg, approximately 3.6 mg / kg, approximately 4.0 mg / kg, approximately 4.8 mg / kg, approximately 5.0 mg / kg, approximately 6.0 mg / kg, approximately 7.2 mg / kg, approximately 8.4 mg / kg, or approximately 10 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0141] In some embodiments, approximately 5-3000 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, approximately 10-500 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, approximately 50-300 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered once every 2 weeks in amounts of about 5 mg, about 10 mg, about 50 mg, about 100 mg, about 126 mg, about 144 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 thereof.

[0142] In some embodiments, after the initial 3-week administration, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks at a dose of about 0.1-10 mg / kg. In some embodiments, after the initial 3-week administration, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks at a dose of about 1-5 mg / kg. In some embodiments, after the initial 3-week administration, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks at a dose of about 1.5-3 mg / kg. In some embodiments, after the initial 3-week administration, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks at a dose of about 2-3 mg / kg. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks after the initial 3-week dosing at approximately 0.1 mg / kg, approximately 0.8 mg / kg, approximately 1 mg / kg, approximately 1.2 mg / kg, approximately 1.5 mg / kg, approximately 2.1 mg / kg, approximately 2.4 mg / kg, approximately 2.7 mg / kg, approximately 3.0 mg / kg, approximately 3.3 mg / kg, approximately 3.6 mg / kg, approximately 4.0 mg / kg, approximately 4.8 mg / kg, approximately 5.0 mg / kg, approximately 6.0 mg / kg, approximately 7.2 mg / kg, approximately 8.4 mg / kg, or approximately 10 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0143] In some embodiments, after the initial 3-week administration, approximately 5-3000 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, after the initial 3-week administration, approximately 10-500 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, after the initial 3-week administration, approximately 50-300 mg of the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks. In some embodiments, the anti-TROP2 antibody-drug conjugate (e.g., ADC1) is administered every 2 weeks after the initial 3-week dosing in amounts of about 5 mg, about 10 mg, about 50 mg, about 100 mg, about 126 mg, about 144 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 thereof.

[0144] In some embodiments, ADC1 is administered at a dose of approximately 1.5-3 mg / kg every 1-4 weeks. In some embodiments, ADC1 is administered at a dose of approximately 2-3 mg / kg every 1-4 weeks. In some embodiments, ADC1 is administered at a dose of approximately 2.1 mg / kg or approximately 2.4 mg / kg every 2 weeks after the initial dose. In some embodiments, ADC1 is administered at a dose of approximately 2.1 mg / kg or approximately 2.4 mg / kg every 2 weeks after the initial dose, which is administered 3 weeks prior.

[0145] In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 1-150 mg / kg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 5-120 mg / kg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 10-100 mg / kg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 20-60 mg / kg per administration. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at a dose of about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 100 mg / kg, about 120 mg / kg, or about 150 mg / kg, or a range (including endpoints) of any two of these values, or any value thereof. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered as a single dose. In some embodiments, it is administered every 1-8 weeks. In some embodiments, it is administered every 1-4 weeks. In some embodiments, it is administered every about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or about 8 weeks. In some implementations, the drug is administered once weekly, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, or once every seven weeks. In some implementations, the drug is administered once every two weeks. In some implementations, the drug is administered once every two weeks after the initial three-week course.

[0146] In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 500-9000 mg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 1000-6000 mg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 1500-3500 mg per administration. In some embodiments, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is about 2000-2800 mg per administration. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered in doses of about 500 mg, about 800 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2400 mg, about 2800 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, about 5000 mg, about 6000 mg, or about 9000 mg, or a range (including endpoints) of any two of these values, or any value thereof. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered as a single dose. In some embodiments, it is administered every 1-8 weeks. In some embodiments, it is administered every 1-4 weeks. In some implementations, the drug is administered once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some implementations, the drug is administered once weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, or every 7 weeks. In some implementations, the drug is administered once every 2 weeks. In some implementations, the drug is administered once every 2 weeks after the initial 3 weeks.

[0147] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 1-150 mg / kg every 1-4 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 5-120 mg / kg every 1-4 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 10-100 mg / kg every 1-4 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 20-60 mg / kg every 1-4 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered once every 1-4 weeks at approximately 1 mg / kg, approximately 5 mg / kg, approximately 10 mg / kg, approximately 20 mg / kg, approximately 30 mg / kg, approximately 40 mg / kg, approximately 50 mg / kg, approximately 60 mg / kg, approximately 100 mg / kg, approximately 120 mg / kg, or approximately 150 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0148] In some embodiments, approximately 500-9000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 1-4 weeks. In some embodiments, approximately 1000-6000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 1-4 weeks. In some embodiments, approximately 1500-3500 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 1-4 weeks. In some embodiments, approximately 2000-2800 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 1-4 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered once every 1-4 weeks in amounts of about 500 mg, about 800 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2400 mg, about 2800 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, about 5000 mg, about 6000 mg, or about 9000 mg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0149] In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 1-150 mg / kg every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 5-120 mg / kg every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 10-100 mg / kg every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered at approximately 20-60 mg / kg every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered once every 2 weeks at approximately 1 mg / kg, approximately 5 mg / kg, approximately 10 mg / kg, approximately 20 mg / kg, approximately 30 mg / kg, approximately 40 mg / kg, approximately 50 mg / kg, approximately 60 mg / kg, approximately 100 mg / kg, approximately 120 mg / kg, or approximately 150 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0150] In some embodiments, approximately 500-9000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, approximately 1000-6000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, approximately 1500-3500 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, approximately 2000-2800 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered once every 2 weeks in amounts of about 500 mg, about 800 mg, about 1000 mg, about 1500 mg, about 2000 mg, about 2400 mg, about 2800 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, about 5000 mg, about 6000 mg, or about 9000 mg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0151] In some embodiments, following the initial 3-week administration, approximately 1-150 mg / kg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 5-120 mg / kg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 10-100 mg / kg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 20-60 mg / kg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks at approximately 1 mg / kg, approximately 5 mg / kg, approximately 10 mg / kg, approximately 20 mg / kg, approximately 30 mg / kg, approximately 40 mg / kg, approximately 50 mg / kg, approximately 60 mg / kg, approximately 100 mg / kg, approximately 120 mg / kg, or approximately 150 mg / kg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0152] In some embodiments, following the initial 3-week administration, approximately 500-9000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 1000-6000 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 1500-3500 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, following the initial 3-week administration, approximately 2000-2800 mg of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks. In some embodiments, the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment (e.g., antibody BsAb-71-N297A) is administered every 2 weeks at approximately 500 mg, approximately 800 mg, approximately 1000 mg, approximately 1500 mg, approximately 2000 mg, approximately 2400 mg, approximately 2800 mg, approximately 3000 mg, approximately 3500 mg, approximately 4000 mg, approximately 4500 mg, approximately 5000 mg, approximately 6000 mg, or approximately 9000 mg, or a range between any two of these values ​​(including the endpoints) or any value thereof.

[0153] In some implementations, approximately 20-60 mg / kg of antibody BsAb-71-N297A is administered every 1-4 weeks. In some implementations, approximately 40 mg / kg of antibody BsAb-71-N297A is administered every 2 weeks. In some implementations, approximately 40 mg / kg of antibody BsAb-71-N297A is administered every 2 weeks after the initial 3-week course.

[0154] In some implementations, approximately 1.5-3 mg / kg of ADC1 and approximately 20-60 mg / kg of antibody BsAb-71-N297A are administered every 1-4 weeks. In some implementations, approximately 2.1 mg / kg of ADC1 and approximately 40 mg / kg of antibody BsAb-71-N297A are administered every 2 weeks. In some implementations, approximately 2.4 mg / kg of ADC1 and approximately 40 mg / kg of antibody BsAb-71-N297A are administered every 2 weeks. In some implementations, after the initial 3-week course of treatment, approximately 2.1 mg / kg of ADC1 and approximately 40 mg / kg of antibody BsAb-71-N297A are administered every 2 weeks. In some implementations, after the initial administration for 3 weeks, ADC1 is administered every 2 weeks at approximately 2.4 mg / kg; and after the initial administration for 3 weeks, antibody BsAb-71-N297A is administered every 2 weeks at approximately 40 mg / kg.

[0155] The term "combination" as used herein refers to a route of administration that includes various situations in which two or more drugs are administered sequentially or simultaneously. In some embodiments, the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment are prepared as a single pharmaceutical composition (e.g., a combination formulation) and administered simultaneously to a patient in need. In some embodiments, the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment are prepared as separate pharmaceutical compositions and administered simultaneously to a patient in need, or administered to a patient in need at different times during a treatment regimen. For example, the anti-TROP2 antibody-drug conjugate may be administered before, after, or alternately with the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment. In some embodiments, the anti-TROP2 antibody-drug conjugate is administered after administration of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment. In this document, the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment are administered to patients in need in a single dose or multiple doses.

[0156] In some implementations, the ADC1 is administered after the antibody BsAb-71-N297A is administered.

[0157] In some implementations, the patient receives one treatment cycle. In some implementations, the patient receives multiple treatment cycles (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 cycles, or a range between any two of these values ​​(including endpoints) or any value thereof). In some implementations, the patient receives treatment until the symptoms are relieved and treatment is no longer required. In some implementations, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, or 7 weeks, or a range between any two of these values ​​(including endpoints) or any value thereof.

[0158] In some embodiments, in the methods and uses for treating tumors provided by the present invention, the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment can be administered via any convenient route, such as by infusion or bolus injection, absorption through epithelial or mucosal membranes (e.g., oral mucosa, rectal and intestinal mucosa), and can be co-administered with other bioactive agents. Therefore, the pharmaceutical composition of the anti-TROP2 antibody-drug conjugate and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment can be administered intravenously, subcutaneously, orally, rectally, parenterally, intracerebrally, vaginally, intraperitoneally, topically (e.g., by powder, ointment, drops, or transdermal patch), orally, orally or via nasal spray. In some embodiments, the route of administration is intravenous (iv) infusion (i.e., intravenous infusion). In some embodiments, an anti-TROP2 antibody-drug conjugate and / or an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment can be formulated into a pharmaceutical composition and administered to a patient in a form suitable for a chosen route of administration, such as parenteral, intravenous (iv), intramuscular, local, or subcutaneous (sc). In some embodiments, the route of administration is intravenous infusion.

[0159] In some embodiments, the anti-TROP2 antibody-drug conjugate, anti-PD-L1 / CD47 bispecific antibody, or antigen-binding fragment is administered via intravenous infusion. In some embodiments, the duration of intravenous infusion is approximately 15 minutes, approximately 30 minutes, approximately 35 minutes, approximately 40 minutes, approximately 45 minutes, approximately 50 minutes, approximately 55 minutes, approximately 60 minutes, approximately 65 minutes, approximately 70 minutes, approximately 75 minutes, approximately 80 minutes, approximately 85 minutes, approximately 90 minutes, approximately 95 minutes, approximately 100 minutes, approximately 105 minutes, approximately 110 minutes, approximately 105 minutes, approximately 120 minutes, approximately 135 minutes, approximately 150 minutes, approximately 165 minutes, approximately 180 minutes, approximately 195 minutes, approximately 210 minutes, approximately 225 minutes, or approximately 240 minutes, or a range (including endpoints) of any two of these values, or any value thereof. In some embodiments, the duration of intravenous infusion is ≥30 minutes. In some implementations, the intravenous infusion duration is ≥60 minutes. In other implementations, the intravenous infusion duration is 30 minutes to 2 hours.

[0160] Examples of tumors include, but are not limited to, triple-negative breast cancer, hormone receptor-positive and HER2-negative (HR+ / HER2-) breast cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), gastric cancer (e.g., gastric adenocarcinoma), esophageal cancer, thymic carcinoma, head and neck tumors (e.g., head and neck squamous cell carcinoma), urothelial carcinoma, ovarian cancer, uterine cancer, endometrial cancer, cervical cancer, fallopian tube cancer, breast cancer, squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), peritoneal cancer, pancreatic cancer, glioblastoma, liver cancer, bladder cancer, urethral cancer, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, colorectal cancer, large intestine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, glioblastoma, medulloblastoma, T-cell lymphoma, melanoma, Kaposi's sarcoma, and hepatobiliary cholangiocarcinoma. In some embodiments, the tumor is an epithelial-derived solid tumor. In some embodiments, the tumor is an advanced solid tumor. In some embodiments, the tumor is a locally advanced or metastatic solid tumor. In some embodiments, the tumor is an advanced solid tumor of epithelial origin. In some embodiments, the tumor is selected from non-small cell lung cancer, gastric adenocarcinoma, esophageal cancer, small cell lung cancer, cervical cancer, or head and neck squamous cell carcinoma. In some embodiments, the tumor is non-squamous non-small cell lung cancer. In some embodiments, the tumor is locally advanced or metastatic non-squamous non-small cell lung cancer.

[0161] In some embodiments, the patient is a patient with a pathologically or cytologically confirmed advanced or metastatic epithelial-derived solid tumor who has failed or is ineligible for standard treatment, or who is intolerant to or refuses standard treatment. In some embodiments, the patient has histologically or cytologically confirmed locally advanced or metastatic non-squamous non-small cell lung cancer that is incurable by radical resection or radiation therapy. In some embodiments, the tumor is driver gene negative and has previously undergone at least one line of systemic chemotherapy. In some embodiments, the tumor is driver gene positive and has previously failed EGFR-TKI therapy for locally advanced or metastatic disease.

[0162] In some implementations, the standard treatment refers to the standard treatment regimen for the tumor recommended by the NCCN guidelines and CSCO guidelines.

[0163] In some implementation schemes, the RECIST v1.1 evaluation criteria are primarily used to assess the antitumor efficacy of ADC1 in combination with or without anti-PD-L1 / CD47 bispecific antibodies or antigen-binding fragments.

[0164] In some embodiments, the combination of the anti-TROP2 antibody-drug conjugate of the present invention and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment exhibits good safety and tolerability. In some embodiments, the combination of the anti-TROP2 antibody-drug conjugate of the present invention and the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment results in improvements selected from at least one of the following: objective response rate (ORR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS).

[0165] In some embodiments, the pharmaceutical composition further includes pharmaceutically acceptable excipients. In some embodiments, pharmaceutically acceptable excipients may comprise antibacterial and / or antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In some embodiments, pharmaceutically acceptable excipients may comprise isotonic agents, such as sugars, polyols (such as mannitol, sorbitol), sodium chloride. In some embodiments, the pharmaceutical composition comprises at least 0.1% of an anti-TROP2 antibody-drug conjugate and / or an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment. The percentage can vary, for example, from about 2% to 90% by weight of a given dosage form. Detailed Implementation

[0166] "Approximately" refers to a typical error range for the corresponding value that is readily known to those skilled in the art. In some embodiments, "approximately" as used herein refers to the described value and its range of ±10%, ±5%, or ±1%.

[0167] "Treatment" refers to therapeutic treatments and preventative or preventative measures aimed at preventing, mitigating, improving, or stopping adverse physiological changes or disorders, such as disease progression, including but not limited to the following, whether detectable or undetectable: symptom relief, reduction of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement, mitigation, reduction, or disappearance of the disease state (whether partial or complete), and prolongation of expected survival without treatment. Patients requiring treatment include those already suffering from the condition or disorder, those susceptible to the condition or disorder, or those needing prevention of the condition or disorder, as well as those who can or are expected to benefit from the application of the antibody or pharmaceutical composition disclosed in this invention for detection, diagnostic procedures, and / or treatment.

[0168] As used herein, the term "effective dose" or "therapeutic effective dose" refers to a dose of a drug, such as an antibody or ADC, sufficient to reduce or improve the severity and / or duration of a condition (e.g., cancer) or one or more of its symptoms; prevent disease progression; induce disease remission; prevent 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 dose of an antibody may 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 dose may improve progression-free survival (PFS), improve overall survival (OS), or reduce the likelihood of recurrence.

[0169] The terms "patient" and "subject" are used interchangeably and refer to any mammal requiring diagnosis, prognosis, or treatment, including but not limited to humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, and especially animals with one or more conditions involving solid tumors. In some implementations, the patient is a human.

[0170] The efficacy indicators for antitumor therapy include objective response rate (ORR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS) as defined by the RECIST 1.1 criteria for evaluating response to solid tumors (Eisenhauer, EA et al., Eur J Cancer. 2009; 45(2):228-247).

[0171] Objective response rate (ORR): refers to the proportion of patients whose tumors shrink to a certain extent and remain so for a certain period of time, including cases of CR and PR.

[0172] Duration of Response (DOR): DOR is defined as the time from the first assessment of objective response to the first assessment of disease progression (PD) or death from any cause prior to PD, reflecting the duration of ORR.

[0173] Disease control rate (DCR): The proportion of patients whose tumors shrink or stabilize for a certain period of time, including cases of CR, PR and SD, also known as clinical benefit rate (CBR).

[0174] Progression-free survival (PFS): the time from the first dose to the occurrence of objective tumor progression or all-cause death (whichever comes first).

[0175] Overall survival (OS): The time from the date of first administration to death from any cause. For patients still alive at the time of analysis, the cutoff date is the date of their last contact.

[0176] Maximum tolerated dose (MTD): MTD is defined as the highest dose level at which DLT is observed in ≤1 / 6 of patients during the DLT assessment period.

[0177] The Eastern Cooperative Oncology Group (ECOG) has developed a simplified activity status scoring system that classifies patients' activity status into 6 levels, from 0 to 5. The ECOG activity status scoring system uses scores of 0, 1, 2, 3, 4, and 5.

[0178] Single-dose pharmacokinetic parameters: C max T max T 1 / 2 CL, Vd, Ke, MRT, AUC (0-τ) AUC (0-∞)。

[0179] Multiple-dose pharmacokinetic parameters: C max,ss C avg,ss C min,ss AUC (0-τ)ss AUC (0-∞)ss T max,ss T 1 / 2,ss CL, V ss Ke, MRT, Accumulation Index (R) ac ), Volatility Index (DF).

[0180] The terms "antibody-drug conjugate" or "ADC" are used interchangeably and refer to a binding protein (such as an antibody or its antigen-binding unit) linked to one or more drugs, which may optionally be a therapeutic agent or a cytotoxic agent. In some embodiments, an ADC includes an antibody, a drug (e.g., a cytotoxic drug), and a connector capable of attaching or conjugating the drug to the antibody. Non-limiting examples of drugs that may be included in an ADC include mitotic inhibitors, antitumor antibiotics, immunomodulators, vectors for gene therapy, alkylating agents, anti-angiogenic agents, antimetabolites, boron-containing agents, chemoprotective agents, hormones, anti-hormonal agents, corticosteroids, photoactive therapeutic agents, oligonucleotides, radionuclides, topoisomerase inhibitors, kinase inhibitors (e.g., TEC-family kinase inhibitors and serine / threonine kinase inhibitors), and radiosensitizers.

[0181] The term "drug-antibody conjugation ratio" or "DAR" refers to the number of drugs (e.g., eczemac) attached to one antibody in 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 binding sites on the antibody. The term DAR may 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, its 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 the composition, i.e., the average number of drug bindings to the antibody, or the average drug-antibody conjugation ratio, its value is selected from about 0 to about 10, or about 2 to about 8. In some embodiments, the average drug-antibody conjugation ratio is about 4 to about 7. In some embodiments, the average drug-antibody conjugation ratio is about 4 to about 5. In other embodiments, the average drug-antibody conjugation ratio is about 6 to about 8, or about 7 to about 8. In some embodiments, the average drug-antibody conjugation ratio is about 6. The DAR value can be represented by p in this paper. The DAR value of ADC can be determined using ultraviolet-visible absorption spectroscopy (UV-Vis), high-performance liquid chromatography-hydrophobic chromatography (HPLC-HIC), high-performance liquid chromatography-reversed-phase chromatography (RP-HPLC), and liquid chromatography-mass spectrometry (LC-MS). These techniques are described in Ouyang, J. Methods Mol Biol, 2013, 1045: pp. 275-83.

[0182] "Antibody" or "antigen-binding fragment" refers to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen. An antibody can be a complete antibody, any antigen-binding fragment, or a single chain thereof. Therefore, the term "antibody" includes any protein or peptide containing at least a portion of an immunoglobulin molecule that has biological activity of binding to an antigen. Antibody and antigen-binding fragments include, but are not limited to, the complementarity-determining region (CDR), heavy chain variable region (VH), light chain variable region (VL), heavy chain constant region (CH), light chain constant region (CL), framework region (FR), or any portion thereof of the heavy chain or light chain or its ligand-binding moiety, or at least a portion of the 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.

[0183] Various substituents are defined as follows.

[0184] "Alkyl" refers to a saturated aliphatic hydrocarbon group; this term includes both straight-chain and branched hydrocarbon groups. C1-C6 alkyl refers to an alkyl group having 1 to 6 carbon atoms, such as alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. "Halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0185] "Stereoisomers" refer to isomers of compounds that have the same atomic bond sequence but different spatial arrangements of atoms. Stereoisomers may have one or more stereocenters, and each center may be present as R or S. Stereoisomers may also be cis-trans isomers. The stereoisomers of compounds provided herein include any one or suitable mixtures of all their diastereomeric, enantiomeric, and cis-trans isomeric forms.

[0186] Pharmaceutically acceptable salts include those produced by compounds with a wide variety of organic and inorganic counterions well known in the art. Exemplary salts include, when the molecule contains acidic functional groups, 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, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethyl, polyamine resins, and tetraalkylammonium salts; and when the molecule contains basic functional groups, 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, and salicylic acid. These salts can usually be prepared by conventional methods by reacting the compound with, for example, a suitable acid or base. Solvates include hydrates.

[0187] Other chemical terms used in this article are used in accordance with the usual usage in the field, such as in The McGraw-Hill Dictionary of Chemical Terms (edited by Parker, S., McGraw-Hill, San Francisco (1985)).

[0188] All publications and patents cited in this document are incorporated herein by reference for all purposes.

[0189] Example

[0190] The following specific embodiments further illustrate the technical solution of the present invention. These specific embodiments do not represent a limitation on the scope of protection of the present invention. Non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the scope of protection of the present invention.

[0191] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0192] The intermediate CB07-Exatecan was prepared according to the preparation method described in patent application WO2022 / 253284. The structure of CB07-Exatecan is shown in Formula IV.

[0193] Antibody BsAb-71-N297A is a bispecific antibody against PD-L1 / CD47, comprising a heavy chain a against PD-L1, a heavy chain b against CD47, a light chain a, and a light chain b. The heavy chain a contains the amino acid sequence shown in SEQ ID NO:16, the heavy chain b contains the amino acid sequence shown in SEQ ID NO:17, and the light chains a and b contain the amino acid sequence shown in SEQ ID NO:18. Its preparation method is described in Example 1 of patent application CN115702931A.

[0194] The antibody hRS9 is an anti-TROP2 antibody, containing a heavy chain as shown in SEQ ID NO:19 and a light chain as shown in SEQ ID NO:20. Its preparation method refers to the preparation and purification of hRS9 antibody in patent application WO2022 / 253284.

[0195] Example 1: Synthesis of ADC1

[0196] Based on the amount of antibody, 3.1 molar equivalents of TCEP (tricarboxyethylphosphine) were added to antibody hRS9, and the pH of the system was adjusted to 7.5 with 1M Tris base. The system was incubated at 25°C for 1 hour for reduction. TCEP was removed by ultrafiltration with 10 mM succinic acid. The thiol antibody value was determined by absorbance measurement. The equivalent amount of free thiol groups was determined by measuring the absorbance at 412 nm of the reaction product of thio groups and DTNB (5,5'-dithiobis(2-nitrobenzoic acid), Aldrich).

[0197] During the coupling reaction, 7 molar equivalents of CB07-Exatecan were added based on the amount of antibody and the content of free sulfhydryl groups. After stirring at 25°C for 1 hour, 0.1M acetylcysteine ​​was added to a final concentration of 2mM, and the reaction was terminated by stirring for another 15 minutes. The mixture was then purified using Capto S ImpAct (cation exchange packing material).

[0198] The DAR value (p) of ADC1 was determined to be approximately 6.

[0199] Example 2: A multicenter, open-label phase IB-II clinical study of the safety, tolerability, pharmacokinetic characteristics, and preliminary efficacy of ADC1 combined with antibody BsAb-71-N297A in patients with advanced solid tumors.

[0200] The research can be broadly divided into two phases:

[0201] The first phase is the dose escalation phase: two dose groups were set up, Group A: 2.1 mg / kg ADC1 + 40 mg / kg antibody BsAb-71-N297A, and Group B: 2.4 mg / kg ADC1 + 40 mg / kg antibody BsAb-71-N297A.

[0202] The second phase is the extended research phase.

[0203] Dosage regimen:

[0204] The order of administration was: 1) antibody BsAb-71-N297A, 2) ADC1.

[0205] ADC1 is administered via intravenous infusion. The first dosing cycle is 21 days (3 weeks), followed by dosing every 2 weeks (Q2W), starting on the first day of each cycle. The infusion should take at least 30 minutes.

[0206] The antibody BsAb-71-N297A is administered via intravenous infusion. The first dosing cycle is 21 days, followed by dosing every 2 weeks (Q2W), starting on the first day of each cycle. An infusion duration of ≥60 minutes is recommended. If no infusion-related reactions are observed, subsequent infusion times can be adjusted to 30 minutes to 2 hours based on the actual clinical situation.

[0207] The baseline weight is the body weight before administration on the first day of the first cycle (C1D1). If the body weight changes by more than 10% from the baseline within 3 days before each administration, the dose is recalculated.

[0208] Selection criteria:

[0209] 1. Age ≥ 18 years old, gender not limited;

[0210] 2. Patients with advanced or metastatic epithelial-derived solid tumors who have been pathologically or cytologically diagnosed, have failed standard treatment or have no standard treatment available, are intolerant to standard treatment, or refuse standard treatment.

[0211] Extended research phase: (Tumor types in the extended cohort can be added or removed based on real-time research results)

[0212] 1) Subjects diagnosed with locally advanced or metastatic non-squamous non-small cell lung cancer by histological or cytological examination, and who are ineligible for radical resection or radiotherapy. They must also meet the following criteria: a. Driver gene-negative patients have previously received at least one line of systemic chemotherapy; b. Driver gene-positive patients have previously received EGFR-TKI therapy for locally advanced or metastatic disease and have failed the treatment.

[0213] 2) For patients with advanced or metastatic epithelial-derived solid tumors who are intolerant to or refuse standard treatment, the following types are preferred: gastric adenocarcinoma, esophageal cancer, small cell lung cancer, cervical cancer, and head and neck squamous cell carcinoma.

[0214] 3. According to RECIST 1.1 criteria, there must be an evaluable tumor lesion during the dose escalation phase and at least one measurable tumor lesion during the dose expansion phase.

[0215] 4. The Eastern Cooperative Oncology Group (ECOG) performance status score requirement is 0 or 1.

[0216] 5. The investigator's assessment of expected survival is ≥12 weeks;

[0217] 6. Possess sufficient organ and bone marrow reserves;

[0218] 7. Female patients of childbearing potential must have a negative pregnancy test within 7 days prior to the first dose and agree to use effective contraception during the study period and for 6 months after the last dose. Male patients must agree to use effective contraception during the study period and for 6 months after the last dose; postmenopausal women must have been without menstruation for at least 12 months to be considered infertile.

[0219] 8. Willing to provide previously archived or fresh tumor tissue samples (this requirement may be waived if there are no previously archived tumor tissue samples and the investigator assesses that re-collecting primary or metastatic tumor tissue samples carries a high risk). Pre-enrollment confirmation of PD-L1 and Trop2 expression levels via immunohistochemistry or other methods is not required; however, the sponsor requires the collection of biopsy or archived tumor tissue samples to determine PD-L1 and Trop2 expression levels, and previous PD-L1 test results are acceptable.

[0220] Exclusion criteria:

[0221] Patients meeting any of the following exclusion criteria will be excluded from the study:

[0222] 1. Within 4 weeks prior to the first administration of the investigational drug, the individual had received treatment with an experimental drug or participated in a clinical trial of a medical device;

[0223] 2. 1) Patients who have received chemotherapy, radical radiotherapy (palliative radiotherapy must be completed within 2 weeks before the first dose), biotherapy, endocrine therapy, immunotherapy, or other anti-tumor treatments within 4 weeks prior to the first dose; 2) For fluorouracil and small molecule targeted drugs, the first dose must be within 5 half-lives of the investigational drug; 3) For nitrosoureas or mitomycin C, the first dose must be within 42 days prior to the first dose of the investigational drug.

[0224] 3. Patients who have received traditional Chinese medicine, proprietary Chinese medicine, or immunomodulatory drugs (including thymopeptides, interferon, interleukin, etc.) with antitumor indications within 2 weeks prior to the first administration of the study drug;

[0225] 4. Prior to the first administration of the study drug, if there are still AEs (CTCAE 5.0) > Grade 1 caused by previous antitumor treatment, excluding toxicities that the investigator judges to pose no safety risk, such as hair loss, Grade 2 peripheral neurotoxicity, etc.

[0226] 5. If a major surgery (excluding diagnostic procedures) is required within 4 weeks prior to the first administration of the investigational drug, or if a major surgery is expected to be required during the study period;

[0227] 6. Individuals who have previously received Trop2-ADC and whose small molecule toxin is a topoisomerase I inhibitor;

[0228] 7. Individuals with a history of allogeneic cell or solid organ transplantation surgery;

[0229] 8. Patients with primary central nervous system tumors or symptomatic central nervous system metastases, those with past or present meningeal metastases, or those with a history of epilepsy. If the brain metastases are stable after treatment, and the lesions have been stable for at least 4 weeks prior to drug administration with no new lesions, or if an asymptomatic lesion is first discovered within 4 weeks and the investigator determines the condition is stable, or if the investigator determines that corticosteroid treatment was discontinued 7 days before the first administration of the study drug, then this is permitted.

[0230] 9. Having other active malignant tumors within 5 years prior to the first dose. This excludes locally cured tumors (e.g., basal cell carcinoma of the skin, squamous cell carcinoma of the skin, superficial bladder cancer, or breast carcinoma in situ, etc.).

[0231] 10. The following cardiovascular diseases occurred within 6 months prior to the first use of medication: symptomatic heart failure of NYHA class 2 or higher, unstable arrhythmia or unstable angina, myocardial infarction requiring treatment, pulmonary embolism, uncontrolled hypertension (defined in this regimen as systolic blood pressure >160 mmHg and / or diastolic blood pressure >100 mmHg after treatment, even with optimal antihypertensive therapy, and clinically significant as assessed by the investigator); QTcF >480 ms as obtained from a 12-lead electrocardiogram (QTcF is calculated using the Fridricia correction formula);

[0232] 11. Subjects with any other serious underlying diseases (e.g., Gilbert's syndrome, uncontrolled diabetes, uncontrolled hypertension, active gastric ulcer, uncontrolled seizures, cerebrovascular events within 3 months prior to first administration, gastrointestinal bleeding, coagulation disorders with severe symptoms or signs) that, in the investigator's opinion, may affect the subject's participation in the study, treatment and follow-up, affect subject compliance, or may lead to complications related to the study drug;

[0233] 12. Received systemic glucocorticoids (prednisone >10 mg / day or equivalent dose of the same drug) or other immunosuppressants within 14 days prior to the first use of the study drug; excluding the following: use of topical, ocular, intra-articular, intranasal and inhaled glucocorticoids, short-term use of glucocorticoids for prophylactic treatment (e.g., prevention of contrast agent allergy);

[0234] 13. Subjects with tuberculosis who are either untreated or currently undergoing treatment, including but not limited to pulmonary tuberculosis; those who have undergone standard anti-tuberculosis treatment and have been confirmed cured by the investigator may be included;

[0235] 14. A serious infection occurred within 4 weeks prior to the first use of medication, or an active infection occurred within 2 weeks prior to the first use of medication;

[0236] 15. Individuals with the following infections: Human Immunodeficiency Virus (HIV) infection; Active Hepatitis B Virus (HBV) infection [HBsAg positive, and HBV-DNA levels >200 IU / ml or 10...]. 3 [Copies / ml]; Hepatitis C virus infected individuals [HCV antibody and viral ribonucleic acid (HCV-RNA) test results positive]; Treponema pallidum antibody positive and RPR / TRUST positive;

[0237] 16. Newly diagnosed thromboembolic events requiring treatment within 6 months (patients with stable lower extremity deep vein thrombosis and patients with port-of-care thrombosis are allowed to be included);

[0238] 17. Uncontrollable pleural effusion, pericardial effusion, or ascites requiring repeated drainage procedures (once a month or more);

[0239] 18. It is known that there is a hypersensitivity reaction or delayed-type hypersensitivity reaction to any component of the research drug;

[0240] 19. Individuals who have a known history of grade ≥3 allergic reactions to macromolecular protein preparations / monoclonal antibodies;

[0241] 20. Known history of mental illness, drug abuse, alcoholism, or drug use that may affect the test results;

[0242] 21. Pregnant or breastfeeding women, or women or men planning to give birth;

[0243] 22. Other circumstances where the researchers deem it unsuitable for participation in this trial.

[0244] Research Objectives and Endpoints

[0245] Part 1: Dosage Elevation Phase

[0246] Research Objective

[0247] Main purpose:

[0248] To evaluate the safety and tolerability of ADC1 combined with antibody BsAb-71-N297A in patients with advanced solid tumors, determine the maximum tolerated dose (MTD), and provide recommended doses for subsequent clinical trials.

[0249] Secondary objective:

[0250] To evaluate the pharmacokinetic (PK) characteristics of ADC1 combined with antibody BsAb-71-N297A in patients with advanced solid tumors under single-dose and multiple-dose administration.

[0251] Evaluation of the immunogenicity of ADC1 combined with antibody BsAb-71-N297A;

[0252] Preliminary evaluation of the antitumor efficacy of ADC1 combined with antibody BsAb-71-N297A;

[0253] To evaluate the relationship between the antitumor efficacy of ADC1 combined with the antibody BsAb-71-N297A and the expression levels of Trop2 and PD-L1 in tumor tissue.

[0254] Study endpoints

[0255] Primary endpoint:

[0256] Tolerability and safety endpoints: vital signs, physical examination, laboratory tests, electrocardiogram, echocardiography, adverse events, dose-limiting toxicity (DLT) events and their incidence, etc.

[0257] Secondary endpoint:

[0258] Pharmacokinetic parameters for single and multiple administrations (mainly including: C60 for a single administration) max T max T 1 / 2 CL, Vd, Ke, MRT, AUC (0-τ) AUC (0-∞) ; Multiple doses of C max,ss C avg,ss C min,ss AUC (0-τ)ss AUC(0-∞)ss T max,ss T 1 / 2,ss CL, V ss Ke, MRT, Accumulation Index (R) ac (Volatility Index DF).

[0259] Immunogenicity assessment indicators: Anti-drug antibody (ADA) / neutralizing antibody (NAb);

[0260] Clinical efficacy indicators: objective response rate (ORR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS);

[0261] Biomarkers: Expression levels of PD-L1 and Trop2 in tumor tissue.

[0262] Part Two: Extended Research Phase

[0263] Research Objective

[0264] Main purpose:

[0265] Further evaluation of the safety and tolerability of ADC1 combined with antibody BsAb-71-N297A in patients with advanced solid tumors;

[0266] The preliminary evaluation of the antitumor efficacy of ADC1 combined with antibody BsAb-71-N297A provides a recommended dose for subsequent clinical trials.

[0267] Secondary objective:

[0268] To evaluate the pharmacokinetic (PK) characteristics of ADC1 combined with antibody BsAb-71-N297A in patients with advanced solid tumors under single-dose and multiple-dose administration.

[0269] Evaluation of the immunogenicity of ADC1 combined with antibody BsAb-71-N297A;

[0270] To evaluate the relationship between the antitumor efficacy of ADC1 combined with antibody BsAb-71-N297A and the expression levels of PD-L1 and Trop2 in tumor tissue.

[0271] Study endpoints

[0272] Primary endpoint:

[0273] Tolerability and safety endpoints: vital signs, physical examination, laboratory tests, electrocardiogram, echocardiography, adverse events, dose-limiting toxicity (DLT) events and their incidence, etc.

[0274] Clinical efficacy indicators: objective response rate (ORR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS).

[0275] Secondary endpoint:

[0276] Pharmacokinetic parameters for single and multiple administrations (mainly including: C60 for a single administration) max T max T 1 / 2 CL, Vd, Ke, MRT, AUC (0-τ) AUC (0-∞) ; Multiple doses of C max,ss C avg,ss C min,ss AUC (0-τ)ss AUC (0-∞)ss T max,ss T 1 / 2,ss CL, V ss Ke, MRT, Accumulation Index (R) ac (Volatility Index DF).

[0277] Immunogenicity assessment indicators: Anti-drug antibody (ADA) / neutralizing antibody (NAb);

[0278] Biomarkers: Expression levels of PD-L1 and Trop2 in tumor tissue.

Claims

1. A method for treating tumors, characterized in that, The method includes administering an effective amount of an anti-TROP2 antibody-drug conjugate and an anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment to a patient in need; the anti-TROP2 antibody-drug conjugate having a structure as shown in Formula I-1 or Formula I-2, or a stereoisomer thereof, or a pharmaceutically acceptable salt or solvate thereof: The formula I-1 is: The formula I-2 is: in Abu is an anti-TROP2 antibody or an antigen-binding fragment; D is a DNA topoisomerase I inhibitor or a salt thereof. For example, the DNA topoisomerase I inhibitor is selected from irinotecan, irinotecan hydrochloride, camptothecin, 9-aminocamptothecin, 9-nitrocamptothecin, 10-hydroxycamptothecin, 9-chloro-10-hydroxycamptothecin, camptothecin derivative SN-38, 22-hydroxyeclipticine, topotecan, letopotecan, belotecone, homosilatecan, 6,8-dibromo-2-methyl-3-[2-(D-pyranoxylamino)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-acrylamide, N-[2-(dimethylamino)ethyl]-4-acrylamide, eczetidine or eczetidine derivatives; n is an integer from 1 to 24; p is approximately 1-10.

2. The method of claim 1, wherein, The D is wherein X 1 and X 2 each independently is C1-C6alkyl, halogen, or -OH; or said C1-C6alkyl is -CH3; or said halogen is F;**is a point of attachment.

3. A method for treating tumors, characterized in that, The method comprises administering to a patient in need thereof an effective amount of an anti-TROP2 antibody drug conjugate and an anti-PD-L1 / CD47 bispecific antibody or antigen binding fragment; the anti-TROP2 antibody drug conjugate has a structure as shown in Formula I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12, I-13, I-14, I-15, I-16, I-17, or I-18, or a stereoisomer thereof or a pharmaceutically acceptable salt or solvate thereof, wherein the Formula I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12, I-13, I-14, I-15, I-16, I-17, and I-18 are: in Abu is an anti-TROP2 antibody or an antigen-binding fragment; n is an integer from 1 to 24; p is approximately 1-10.

4. The method according to any one of claims 1-3, characterized in that, n is an integer between 4 and 12, or an integer between 4 and 8, or n is 4, or n is 8.

5. The method according to any one of claims 1-4, characterized in that, p is approximately 2-8, or p is approximately 4-8, or p is approximately 4-5, or p is approximately 6-8, or p is approximately 7-8, or p is approximately 4-6, or p is approximately 5-7, or p is approximately 5.5-6.5, or p is approximately 4, or p is approximately 6.

6. The method according to any one of claims 1-5, characterized in that, The anti-TROP2 antibody or antigen-binding fragment comprises a heavy chain as shown in SEQ ID NO:19 and a light chain as shown in SEQ ID NO:

20.

7. The method according to any one of claims 1-6, characterized in that, The anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment comprises a variable region a that specifically binds to PD-L1 and a variable region b that specifically binds to CD47; the variable region a comprises at least one or more of VHa CDR1 as shown in SEQ ID NO:1, VHa CDR2 as shown in SEQ ID NO:2, VHa CDR3 as shown in SEQ ID NO:3, VLa CDR1 as shown in SEQ ID NO:4, VLa CDR2 as shown in SEQ ID NO:5, and VLa CDR3 as shown in SEQ ID NO:6; the variable region b comprises at least one or more of VHb CDR1 as shown in SEQ ID NO:7, VHb CDR2 as shown in SEQ ID NO:8, VHb CDR3 as shown in SEQ ID NO:9, VLb CDR1 as shown in SEQ ID NO:4, VLb CDR2 as shown in SEQ ID NO:5, and VLb CDR3 as shown in SEQ ID NO:

6.

8. The method according to any one of claims 1-7, characterized in that, The anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment has a greater affinity for PD-L1 than for CD47.

9. The method according to any one of claims 1-8, characterized in that, The variable region a includes VHa CDR1 as shown in SEQ ID NO:1, VHa CDR2 as shown in SEQ ID NO:2, VHa CDR3 as shown in SEQ ID NO:3, VLa CDR1 as shown in SEQ ID NO:4, VLa CDR2 as shown in SEQ ID NO:5, and VLa CDR3 as shown in SEQ ID NO:6; the variable region b includes VHb CDR1 as shown in SEQ ID NO:7, VHb CDR2 as shown in SEQ ID NO:8, VHb CDR3 as shown in SEQ ID NO:9, VLb CDR1 as shown in SEQ ID NO:4, VLb CDR2 as shown in SEQ ID NO:5, and VLb CDR3 as shown in SEQ ID NO:

6.

10. The method according to any one of claims 7-8, characterized in that, The variable region a comprises a heavy chain variable region VHa and a light chain variable region VLa; wherein... The VHa comprises an amino acid sequence as shown in SEQ ID NO:10, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:10, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:10; and / or The VLa contains an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:

12.

11. The method according to any one of claims 7-9, characterized in that, The variable region b comprises the heavy chain variable region VHb and the light chain variable region VLb; wherein... The VHb comprises an amino acid sequence as shown in SEQ ID NO:11, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:11, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:11; and / or The VLb contains an amino acid sequence as shown in SEQ ID NO:12, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:

12.

12. The method according to any one of claims 7-10, characterized in that, The anti-PD-L1 / CD47 bispecific antibody further comprises a heavy chain constant region CHa and a heavy chain constant region CHb, wherein the heavy chain constant region CHa is connected to the heavy chain variable region VHa to form heavy chain a, and the heavy chain constant region CHb is connected to the heavy chain variable region VHb to form heavy chain b.

13. The method according to any one of claims 7-10, wherein the heavy chain constant region CHa and / or the heavy chain constant region CHb contains an amino acid mutation of N297A, wherein the amino acid position is Eu numbered.

14. The method of any one of claims 7-10, wherein the heavy chain constant region CHa comprises an amino acid sequence as shown in SEQ ID NO:13, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:13, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:13; and / or The heavy chain constant region CHb contains an amino acid sequence as shown in SEQ ID NO:15, or an amino acid sequence that is at least 80% or at least 90% identical to the amino acid sequence shown in SEQ ID NO:15, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:

15.

15. The method according to any one of claims 7-10, wherein the anti-PD-L1 / CD47 bispecific antibody comprises heavy chain a, heavy chain b, light chain a, and light chain b; wherein, The heavy chain a comprises an amino acid sequence as shown in SEQ ID NO:16, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:16, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:16; and / or The heavy chain b comprises an amino acid sequence as shown in SEQ ID NO:17, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:17, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:17; and / or Light chains a and b contain the same amino acid sequence.

16. The method according to any one of claims 7-15, wherein the light chains a and b comprise an amino acid sequence as shown in SEQ ID NO:18, or an amino acid sequence having at least 80% or at least 90% identity with the amino acid sequence shown in SEQ ID NO:18, or an amino acid sequence having one or more conserved amino acid substitutions compared to the amino acid sequence shown in SEQ ID NO:

18.

17. The method according to any one of claims 1-16, characterized in that, The dosage of the anti-TROP2 antibody-drug conjugate is approximately 0.1-10 mg / kg; or, the dosage of the anti-TROP2 antibody-drug conjugate is approximately 1-5 mg / kg; or, the dosage of the anti-TROP2 antibody-drug conjugate is approximately 1.5-3 mg / kg; or, the dosage of the anti-TROP2 antibody-drug conjugate is approximately 2-3 mg / kg; or, the dosage of the anti-TROP2 antibody-drug conjugate is approximately 2.1 mg / kg or approximately 2.4 mg / kg.

18. The method according to any one of claims 1-16, wherein the dose of the anti-TROP2 antibody-drug conjugate is about 5-3000 mg per administration; or, the dose of the anti-TROP2 antibody-drug conjugate is about 10-500 mg per administration; or, the dose of the anti-TROP2 antibody-drug conjugate is about 50-300 mg per administration.

19. The method according to any one of claims 1-18, characterized in that, The dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is approximately 1-150 mg / kg per administration; or, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is approximately 10-100 mg / kg per administration; or, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is approximately 20-60 mg / kg per administration; or, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is approximately 40 mg / kg per administration.

20. The method according to any one of claims 1-19, wherein the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is about 500-9000 mg per administration; or, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is about 1500-3500 mg per administration; or, the dose of the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is about 2000-2800 mg per administration.

21. The method according to any one of claims 1-20, characterized in that, The anti-TROP2 antibody-drug conjugate and / or anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment may be administered once every 1-8 weeks, or once every 1-4 weeks; for example, once every 2 weeks, or once every 2 weeks after the first administration 3 weeks later.

22. The method according to any one of claims 1-21, characterized in that, The anti-TROP2 antibody-drug conjugate is administered at a dose of approximately 1.5-3 mg / kg every 1-4 weeks, for example, every 2 weeks, or every 2 weeks after the initial 3 weeks of administration; the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is administered at a dose of approximately 20-60 mg / kg every 1-4 weeks, for example, every 2 weeks, or every 2 weeks after the initial 3 weeks of administration.

23. The method according to any one of claims 1-22, characterized in that, The anti-TROP2 antibody-drug conjugate is administered at a dose of approximately 2.1 mg / kg or approximately 2.4 mg / kg every 2 weeks; the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is administered at a dose of approximately 40 mg / kg every 2 weeks.

24. The method according to any one of claims 1-23, characterized in that, The anti-TROP2 antibody-drug conjugate is administered at a dose of approximately 2.1 mg / kg or approximately 2.4 mg / kg, with the first administration followed by administration every 2 weeks for 3 weeks; the anti-PD-L1 / CD47 bispecific antibody or antigen-binding fragment is administered at a dose of approximately 40 mg / kg, with the first administration followed by administration every 2 weeks for 3 weeks.

25. The method according to any one of claims 1-24, characterized in that, The tumors are selected from triple-negative breast cancer, hormone receptor-positive and HER2-negative (HR+ / HER2-) breast cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), gastric cancer (e.g., gastric adenocarcinoma), esophageal cancer, thymic carcinoma, head and neck tumors (e.g., head and neck squamous cell carcinoma), urothelial carcinoma, ovarian cancer, uterine cancer, endometrial cancer, cervical cancer, fallopian tube cancer, breast cancer, squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), peritoneal cancer, pancreatic cancer, glioblastoma, liver cancer, bladder cancer, urethral cancer, hepatoma, colon cancer, rectal cancer, colorectal cancer, colorectal cancer, large intestine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, glioblastoma, medulloblastoma, T-cell lymphoma, melanoma, Kaposi's sarcoma, and hepatobiliary carcinoma.

26. The method according to any one of claims 1-25, characterized in that, The tumor is a solid tumor of epithelial origin.

27. The method according to any one of claims 1-26, characterized in that, The tumor was an advanced solid tumor.

28. The method according to any one of claims 1-27, characterized in that, The tumor is a solid tumor of epithelial origin.

29. The method according to any one of claims 1-28, characterized in that, The tumors were selected from non-small cell lung cancer, gastric adenocarcinoma, esophageal cancer, small cell lung cancer, cervical cancer, and head and neck squamous cell carcinoma.

30. The method according to any one of claims 1-29, characterized in that, The tumor is non-squamous non-small cell lung cancer.

31. The method according to any one of claims 1-30, characterized in that, The tumor is locally advanced or metastatic non-squamous non-small cell lung cancer.

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