Pharmaceutical composition containing Anti-nectin4 antibody-drug conjugate and Anti-PD-1 antibody and use thereof

Abstract

A pharmaceutical composition containing an anti-Nectin4 antibody-drug conjugate and an anti-PD-1 antibody and a use thereof in preventing or treating cancer. The pharmaceutical composition has good safety, and has a stronger anti-tumor effect than using a drug alone. The pharmaceutical composition shows a synergistic tumor inhibitory effect. The present invention also relates to a kit of parts containing the pharmaceutical composition.

Description

Drug combination of anti-nectin4 antibody drug conjugate and anti-PD-1 antibody and use thereof Technical Field

[0001] The present invention relates to the field of medicine. Specifically, the present invention relates to a drug combination comprising an anti-nectin4 antibody-drug conjugate and an anti-programmed death-1 (PD-1) antibody or an antigen-binding fragment thereof that targets PD-1, wherein the drug combination is used to prevent or treat cancer. Background Art

[0002] Nectin-4 (also known as "poliovirus receptor-like molecule 4 (PVRL4)") is a type I transmembrane glycoprotein with a molecular weight of approximately 66 kD. It is a member of the nectin family in the immunoglobulin superfamily. Its extracellular domain is composed of three Ig-like domains (VCC type) and participates in the formation and maintenance of adherens junctions together with cadherins.

[0003] Nectin-4 is present at high levels in normal embryonic and fetal tissues, decreases in adulthood, and has a limited distribution in healthy tissues. It is highly expressed in a variety of tumor cells, such as colon, ovarian, cervical, urothelial, breast, lung, gastric, and head and neck cancers, primarily promoting tumor cell proliferation, differentiation, migration, and invasion by activating the PI3K / Akt pathway. Therefore, targeting nectin-4 is becoming an effective strategy for treating cancers expressing nectin-4.

[0004] Anti-Nectin4 antibody-drug conjugate (anti-Nectin4-ADC) is composed of a monoclonal antibody targeting the specific antigen Nectin-4 and a small molecule cytotoxic drug connected by a linker (also called a "connector"). It combines the powerful killing effect of traditional small molecule chemotherapy with the tumor targeting of antibody drugs. It consists of three main parts: an antibody responsible for selectively recognizing the cancer cell surface antigen Nectin-4, a drug payload responsible for killing cancer cells, and a linker connecting the antibody and the payload.

[0005] Although antibody-drug conjugates (ADCs) have become a popular drug class for the treatment of hematological malignancies and solid tumors, extensive preclinical and clinical studies have shown that the duration of objective responses or clinical spillover produced by ADCs as monotherapy in tumor treatment using ADCs is still limited. Therefore, the combination of ADCs with other anticancer drugs has become an important direction for ADC drug development.

[0006] PD-1 is a key immune checkpoint receptor expressed by activated T and B cells and mediates immune suppression (Yao S, Zhu Y and Chen L., Advances in targeting cell surface signaling molecules for immune modulation. Nat Rev Drug Discov, 2013, 12(2): 130-146). Two cell surface glycoprotein ligands for PD-1 have been identified, namely programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2). They are expressed on antigen-presenting cells and many human cancers. Existing research results have shown that binding to PD-1 can lead to T cell apoptosis, immune anergy, T cell "exhaustion" and IL-10 secretion. Studies have shown that blocking the binding of PD-1 to its ligand can restore T cell function in cancer patients. Monoclonal antibodies targeting PD-1 have been described, for example, toripalimab, pembrolizumab or pidilizumab. After binding to PD-1 on T lymphocytes, the anti-PD-1 monoclonal antibody can inhibit the binding of PD-1 to its ligand, thereby promoting T lymphocyte activation, proliferation and production of immune-activating cytokines such as IL-2, and relieving PD-1's inhibition of T lymphocyte immune surveillance with anti-tumor activity.

[0007] The biological behavior of most tumors is not dominated by a single signal transduction pathway, but by multiple signal transduction pathways working together. In some cases, drugs with different mechanisms of action can be used in combination. However, any combination of drugs that have different mechanisms of action but work in similar fields does not necessarily produce a combination with beneficial effects. Therefore, although there is indeed a demand for combination drug regimens and products for different signal transduction pathways in the prior art, in view of the complexity of the tumorigenesis mechanism, the unpredictability of the interactions between different drugs and other factors, it is still a major challenge in the medical field to find feasible and combination drug regimens and products that can bring better effects than single drugs (reducing the dose of a single drug, improving the incidence and / or severity of adverse events (AEs) during treatment, and / or working in a synergistic manner, etc.).

[0008] Summary of the Invention

[0009] The present inventors surprisingly found that the drug combination of the present application can prevent and / or treat cancer in a manner of synergistic effect and / or improving the incidence and / or severity of adverse events (AEs) during treatment.

[0010] The present invention provides a drug combination comprising an anti-nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof and an anti-PD-1 antibody or an antigen-binding fragment thereof, and use of the combination for preventing or treating cancer.

[0011] Specifically, the present invention relates to the following aspects.

[0012] In a first aspect, the present invention provides a pharmaceutical combination comprising

[0013] (i) an anti-Nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof; and

[0014] (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0015] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof in the pharmaceutical combination of the present invention comprises

[0016] (i) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, and HCDR3 shown in SEQ ID NO: 13; and the light chain variable region comprises LCDR1 shown in SEQ ID NO: 14, LCDR2 shown in SEQ ID NO: 15, and LCDR3 shown in SEQ ID NO: 16; or

[0017] (ii) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1 of SEQ ID NO: 21, HCDR2 of SEQ ID NO: 22, and HCDR3 of SEQ ID NO: 23; and the light chain variable region comprises LCDR1 of SEQ ID NO: 24, LCDR2 of SEQ ID NO: 25, and LCDR3 of SEQ ID NO: 26;

[0018] Preferably, the anti-PD-1 antibody or antigen-binding fragment thereof comprises

[0019] (i) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the sequence of SEQ ID NO: 17, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and the light chain variable region comprises the sequence of SEQ ID NO: 18, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto; or

[0020] (ii) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the sequence of SEQ ID NO:27, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and the light chain variable region comprises the sequence of SEQ ID NO:28, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto;

[0021] More preferably, the anti-PD-1 antibody comprises

[0022] (i) SEQ ID NO: 19 or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO: 20 or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto; or

[0023] (ii) SEQ ID NO: 29 or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO: 30 or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto.

[0024] In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment in the pharmaceutical combination of the present invention is an IgG antibody; preferably, it is a human IgG antibody; more preferably, it is a human IgG1 or human IgG4 antibody; wherein the antigen-binding fragment is Fab, Fab', F(ab')2, Fv, single-chain Fv, single-chain Fab.

[0025] In some embodiments, the anti-nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof in the drug combination of the present invention has a molecular formula Ab-[L-CTD]m, wherein Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof, L represents a linker, CTD represents a drug, and m represents the average number of drug connections relative to each molecule of Ab; preferably, CTD is a cytotoxic drug, preferably, CTD is one or more selected from the following: microtubule inhibitors MMAE, DM1, DM4, Tublysin, amanitin, calicheamicin, eribulin and derivatives of the drugs; topoisomerase inhibitors SN38, exitecan and derivatives of the drugs; DNA binders PBD, doxorubicin and derivatives of the drugs; preferably, m is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, further preferably 3.8-4.2, further preferably 3.9-4.1, and particularly preferably 4.0.

[0026] In some embodiments, the anti-nectin-4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof in the pharmaceutical combination of the present invention has the molecular formula Ab-[L-CTD]m, wherein Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof, and the Ab comprises a heavy chain variable region and a light chain variable region, and the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 1, HCDR2 shown in SEQ ID NO: 2, and HCDR3 shown in SEQ ID NO: 3; and the light chain variable region comprises LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6;

[0027] Preferably, the Ab comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the sequence shown in SEQ ID NO: 7, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereof, and the light chain variable region comprises the sequence shown in SEQ ID NO: 8, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereof;

[0028] For example, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8 or SEQ ID NO: 10;

[0029] More preferably, the Ab comprises:

[0030] (i) the heavy chain variable region amino acid sequence shown in SEQ ID NO: 7; and the light chain variable region amino acid sequence shown in SEQ ID NO: 8; or

[0031] (ii) the heavy chain variable region amino acid sequence shown in SEQ ID NO: 9; and the light chain variable region amino acid sequence shown in SEQ ID NO: 10;

[0032] Preferably, the Ab is an IgG antibody; more preferably, it is a human IgG antibody; most preferably, it is a human IgG1 or human IgG4 antibody; wherein the antigen-binding fragment is Fab, Fab', F(ab')2, Fv, single-chain Fv, single-chain Fab.

[0033] In some embodiments, the anti-nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof in the pharmaceutical combination of the present invention has a structure as shown in Formula Ia and / or Ib below: [0033.1] [Corrected 08.04.2025 according to Rule 26]

[0034] Wherein: Ab is an anti-nectin-4 antibody or an antigen-binding fragment thereof;

[0035] Ar' is any one selected from the following: a substituted or unsubstituted C6-C10 arylene group and a substituted or unsubstituted 5-12 membered heteroarylene group, wherein the substitution refers to the replacement of hydrogen atoms on the group by one or more substituents, and the substituents are selected from any one of the following: halogen (F, Cl, Br or I), haloalkyl (e.g., haloC1-C6 alkyl, preferably haloC1-C4 alkyl, such as trifluoromethyl) and alkoxy (e.g., C1-C6 alkoxy, preferably C1-C4 alkoxy, such as methoxy);

[0036] L1 is -O(CH2CH2O)n- connected to the Ar' group, wherein n is selected from any integer in the range of 1-24, preferably any integer in the range of 1-10, more preferably any integer in the range of 3-5;

[0037] L2 is an enzymatically cleavable fragment, such as a dipeptide, tripeptide, or tetrapeptide, or a combination thereof with a self-releasing structural fragment (i.e., a polypeptide fragment consisting of 2-4 amino acids, or a combination thereof with a self-releasing structural fragment), such as Val-Ala, Val-Ala-PAB, Val-Cit, Val-Cit-PAB, Phe-Lys-PAB, Ala-Ala-Ala, Gly-Gly-Phe-Gly (GGFG), and MAC glucuronide phenol.

[0038] In some embodiments, L2-CTD is VcMMAE, GGFG-Dxd or VC-seco-DUBA; preferably, when Ar' is a substituted or unsubstituted 5-12 membered heteroarylene group, the heteroatom is N; preferably, Ar' is a substituted or unsubstituted C6 arylene group or a substituted or unsubstituted 6 membered heteroarylene group; more preferably, the antibody drug conjugate or a pharmaceutically acceptable salt thereof has the following structure:

[0039] Conjugate ADC-1:

[0040] Conjugate ADC-2:

[0041] Conjugate ADC-3:

[0042] Conjugate ADC-4:

[0043] Conjugate ADC-5:

[0044] Conjugate ADC-6:

[0045] Conjugate ADC-7:

[0046] In some embodiments, a pharmaceutical combination of the present invention is provided, wherein:

[0047] (i) The anti-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof is represented by the following formula:

[0048] Wherein, the anti-nectin-4 antibody is as described above,

[0049] N represents the average number of drug linkages per anti-nectin-4 antibody molecule, which is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, further preferably 3.8-4.2, further preferably 3.9-4.1, and particularly preferably 4.0; and

[0050] (ii) The anti-PD-1 antibody or antigen-binding fragment thereof is an anti-PD-1 antibody or antigen-binding fragment thereof selected from toripalimab or pembrolizumab.

[0051] In a second aspect, the present invention provides a use of the drug combination described in the first aspect for preparing a drug for preventing or treating cancer. Preferably, the cancer is any one selected from the following: urothelial carcinoma, bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, gastric cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute lymphocytic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, prostate cancer, colorectal cancer, colon cancer, glioma, mesothelioma.

[0052] In a third aspect, the present invention provides a kit comprising the pharmaceutical combination according to the first aspect of the present invention, preferably the kit is in the form of a pharmaceutical dosage unit.

[0053] Other aspects and embodiments of the present invention will become apparent by reference to the detailed description which follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The preferred embodiments of the present invention described in detail below will be better understood when read in conjunction with the following drawings. For the purpose of illustrating the present invention, the drawings show presently preferred embodiments. However, it should be understood that the present invention is not limited to the precise arrangements and means of the embodiments shown in the drawings.

[0055] Figure 1: Shows the effect of anti-Nectin4-ADC combined with PD-1 antibody pembrolizumab or administered alone on tumor volume in tumor-bearing mice.

[0056] Figure 2: Shows the effect of anti-Nectin4-ADC combined with PD-1 antibody pembrolizumab or administered alone on the body weight of tumor-bearing mice.

[0057] Figure 3: Shows the effect of anti-Nectin4-ADC in combination with PD-1 antibody Toripalimab or administration alone on tumor volume in tumor-bearing mice.

[0058] Figure 4 shows the effect of anti-Nectin4-ADC combined with PD-1 antibody Toripalimab or administered alone on the body weight of tumor-bearing mice. DETAILED DESCRIPTION

[0059] Before describing the present invention in detail, it should be understood that the present invention is not limited to the specific methods and experimental conditions in this specification, because the methods and conditions can be varied. In addition, the terminology used herein is only for describing specific embodiments and is not intended to be limiting.

[0060] I. Definition

[0061] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For the purposes of the present invention, the following terms are defined below.

[0062] The term "about" when used in conjunction with a numerical value is meant to encompass the numerical value within a range having a lower limit that is 10% less than the specified numerical value and an upper limit that is 10% greater than the specified numerical value.

[0063] The term "and / or" when used to link two or more alternatives should be understood to mean any one of the alternatives or any two or more of the alternatives.

[0064] As used herein, the terms "comprising" or "including" are intended to include the recited elements, integers, or steps, but do not exclude any other elements, integers, or steps. In this document, when the terms "comprising" or "including" are used, unless otherwise indicated, the context of consisting of the recited elements, integers, or steps is also encompassed. For example, when reference is made to an antibody variable region "comprising" a specific sequence, it is intended to encompass an antibody variable region consisting of that specific sequence.

[0065] The term "antibody" is used in the broadest sense to refer to a protein that contains an antigen binding site and encompasses natural antibodies and artificial antibodies of various structures, including but not limited to intact antibodies and antigen-binding fragments of antibodies.

[0066] The terms "whole antibody," "full-length antibody," "complete antibody," and "intact antibody" are used interchangeably herein to refer to a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions can be further divided into hypervariable regions (complementarity determining regions (CDRs) interspersed with more conserved regions (framework regions (FRs)). Each VH and VL is composed of three CDRs and four FRs, arranged from amino terminus to carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The constant region is not directly involved in the binding of the antibody to the antigen, but exhibits various effector functions. In a given VH or VL amino acid sequence, the precise amino acid sequence boundaries of each CDR can be determined using any one or a combination of many well-known schemes, including, for example, the Chothia numbering scheme (Chothia et al., "Canonical structures for the hypervariable regions of immunoglobulins", Journal of Molecular Biology, 196, 901-917 (1987)); the Kabat numbering scheme (Kabat et al., Sequences of Proteins of Immunological Interest, 4th Edition, US Department of Health and Human Services, National Institutes of Health, 1994); Institutes of Health (1987)), AbM (University of Bath) and Contact (University College London); North numbering scheme (North et al., A New Clustering of Antibody CDR Loop Conformations", Journal of Molecular Biology, 406, 228-256 (2011)). The CDRs of anti-nectin4 in the anti-nectin4 antibody drug conjugate of the present invention and the CDRs of the anti-PD-1 antibody used in combination with the anti-nectin4 antibody drug conjugate can be determined according to any scheme or combination thereof in the art and human evaluation to determine the boundaries. In one embodiment, the CDRs of the antibody of the present invention are CDR sequences defined according to the Kabat numbering scheme.

[0067] Although CDR is different between antibodies, only a limited number of amino acid positions in CDR are directly involved in antigen binding. Using at least two of the Kabat, Chothia, AbM and Contact methods, the minimum overlapping region can be determined, thereby providing a "minimum binding unit" for antigen binding. The minimum binding unit can be a sub-portion of a CDR. As those skilled in the art will appreciate, the residues of the remainder of the CDR sequence can be determined by the structure and protein folding of the antibody. Therefore, the present invention also contemplates variants of any CDR provided herein. For example, in a variant of a CDR, the amino acid residues of the minimum binding unit can remain unchanged, while the remaining CDR residues defined according to Kabat or Chothia can be replaced by conservative amino acid residues.

[0068] The term "antigen-binding fragment" is a portion or segment of an intact or complete antibody that has fewer amino acid residues than the intact or complete antibody and that is capable of binding to an antigen or competing with the intact antibody (i.e., the intact antibody from which the antigen-binding fragment is derived) for antigen binding. Antigen-binding fragments can be prepared by recombinant DNA technology, or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, single-chain Fv, diabodies, and single-domain antibodies (sdAbs). The Fab fragment is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains. For example, Fab fragments can be obtained by digesting an intact antibody with papain. In addition, digesting an intact antibody with pepsin below the disulfide bonds in the hinge region produces F(ab')2, a dimer of Fab', a divalent antibody fragment. F(ab')2 can be reduced under neutral conditions by breaking the disulfide bonds in the hinge region, thereby converting the F(ab')2 dimer into a Fab' monomer. The Fab' monomer is essentially a Fab fragment with a hinge region (for a more detailed description of other antibody fragments, see: Fundamental Immunology, edited by W.E. Paul, Raven Press, NY (1993)). The Fv fragment consists of the VL and VH domains of a single antibody arm. In addition, although the two domains of the Fv fragment, VL and VH, are encoded by independent genes, recombinant methods can be used to connect them via a synthetic linker peptide that enables the two domains to be produced as a single protein chain, in which the VL and VH regions are paired to form a single-chain Fv. The antibody fragment can be obtained by chemical methods, recombinant DNA methods, or protease digestion.

[0069] The term "monoclonal antibody" refers to polypeptides, including antibodies and antigen-binding fragments, that have substantially identical amino acid sequences or are derived from the same genetic source. The term also includes preparations of antibody molecules having a single molecular composition. A monoclonal antibody exhibits a single binding specificity and affinity for a particular epitope.

[0070] The term "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human CDRs and amino acid residues from human FRs. In some embodiments, the humanized antibody comprises all or substantially all CDRs corresponding to those of non-human antibodies and all or substantially all FR regions corresponding to those of human antibodies. The humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody (e.g., a non-human antibody) refers to an antibody that has undergone humanization.

[0071] The term "specific binding" or the like means that an antibody forms a complex with an antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, surface plasmon resonance assays, MSD assays (Estep, P. et al., High throughput solution-based measurement of antibody-antigen affinity and epitope binning, MAbs, 2013. 5(2): p. 270-278), and the like.

[0072] In the context of two or more nucleic acid sequences or peptide sequences, the term "% identity" refers to the percent identity between sequences. If two sequences have identical amino acid sequences or nucleotide sequences in the compared region, they are "identical." When comparing and alignment within a comparison window or a specified region to seek the maximum correspondence measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection, if the two sequences have an amino acid residue or nucleotide with a specified percent identity (i.e., 60% identity, optionally 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% identity in the specified region or over the entire sequence when there is no specification), the two sequences are "substantially identical." The calculation of sequence identity between sequences is as follows.

[0073] To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of the first and second amino acid or nucleic acid sequences for optimal alignment or non-homologous sequences can be discarded for comparison purposes). In a preferred embodiment, for comparison purposes, the length of the reference sequence being aligned is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide at the corresponding position in the second sequence, then the molecules are identical at that position.

[0074] Mathematical algorithms can be used to compare sequences and calculate percent identity between two sequences. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needlema and Wunsch ((1970) J. Mol. Biol. 48:444-453) algorithm (available at http: / / www.gcg.com), which has been integrated into the GAP program in the GCG software package. The Blossum 62 matrix or the PAM250 matrix is ​​used, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 is used, and a length weight of 1, 2, 3, 4, 5, or 6 is used. In another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at http: / / www.gcg.com). A particularly preferred parameter set (and the one that should be used unless otherwise stated) is the Blossum 62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5.

[0075] The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of E. Meyers and W. Miller, (1989) CABIOS, 4: 11-17), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weighted remainder table, a gap length penalty of 12, and a gap penalty of 4.

[0076] Additionally or alternatively, one can further use the nucleic acid sequences and protein sequences described herein as a "query sequence" to perform searches against public databases to, for example, identify other family member sequences or related sequences.

[0077] The term "immune checkpoint" refers to a class of inhibitory signaling molecules present in the immune system that prevent tissue damage by regulating the persistence and intensity of immune responses in peripheral tissues and participate in maintaining tolerance to self-antigens (Pardoll DM., The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer, 2012, 12(4): 252-264). Studies have found that one of the reasons why tumor cells can evade the immune system in the body and proliferate uncontrollably is that they utilize the inhibitory signaling pathway of immune checkpoints, thereby inhibiting the activity of T lymphocytes, making it impossible for T lymphocytes to effectively exert their killing effect on tumors (Yao S, Zhu Y and Chen L., Advances in targeting cell surface signaling molecules for immune modulation. Nat Rev Drug Discov, 2013, 12(2): 130-146). Immune checkpoint molecules include but are not limited to programmed death 1 (PD-1), PD-L1, PD-L2, cytotoxic T lymphocyte antigen 4 (CTLA-4), LAG-3, TIM-3, and proteins with T cell immunoglobulin and ITIM domains (TIGIT).

[0078] The term "drug combination" refers to a non-fixed combination product or a fixed combination product, including but not limited to a kit and a pharmaceutical composition. The term "non-fixed combination" means that the active ingredients (e.g., (i) an anti-Nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof, and (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof) are administered to a patient as separate entities simultaneously, without specific time restrictions, or sequentially at equal or different time intervals, wherein such administration provides preventively or therapeutically effective levels of the two active agents in the patient's body. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment and the anti-Nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof used in the drug combination are administered at levels no greater than when they are used alone. The term "fixed combination" means that the two active agents are administered to the patient simultaneously in the form of a single entity. The dosage and / or time interval of the two active agents are preferably selected so that the combined use of the parts can produce an effect greater than that achieved by using either component alone when treating a disease or condition. Each component can be in the form of a separate formulation, and the formulation forms can be the same or different.

[0079] The terms "administer" and "administer" are used interchangeably and refer to the physical introduction of each active ingredient in the drug combination of the present invention into an individual using any of a variety of methods and delivery systems known to those skilled in the art. The route of administration of each active ingredient in the drug combination of the present invention includes oral, intravenous (e.g., infusion (also known as drip) or injection), intramuscular, subcutaneous, intraperitoneal, spinal, local or other parenteral administration routes. The term "parenteral administration" refers to the mode of administration beyond gastrointestinal and topical administration, typically by intravenous, and includes, but is not limited to, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardial, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. Accordingly, each active ingredient in the drug combination of the present invention can be formulated into capsules, tablets, injections (including infusions or injections), syrups, sprays, lozenges, liposomes or suppositories etc.

[0080] The term "dose" refers to the amount of a drug required to elicit a therapeutic effect. Unless otherwise indicated, the dose is relative to the amount of the drug in free form. If the drug is in the form of a pharmaceutically acceptable salt, the amount of drug is proportionally increased compared to the amount of the drug in free form. For example, the dose will be stated on the product packaging or product information sheet.

[0081] The term "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0082] The term "cancer" refers to a disease characterized by the abnormal cell proliferation of rapid and uncontrolled growth. Cancer cells can spread to other parts of the body locally or through the bloodstream and lymphatic system. Cancer includes but is not limited to solid tumors and hematological malignancies, preferably solid tumors. The example of various cancers includes but is not limited to urothelial carcinoma, bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, gastric cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute lymphocytic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, prostate cancer, colorectal cancer, colon cancer, glioma, mesothelioma. The cancer is preferably advanced cancer, recurring and / or refractory cancer or a cancer resistant to chemotherapy, more preferably advanced solid tumors, such as (diagnosed through histology or cytology) inoperable or metastatic advanced solid tumors.

[0083] The term "inhibit" refers to a decrease in a certain parameter (e.g., nectin-4 activity and / or PD-1 activity) caused by a given molecule (e.g., (i) an anti-nectin-4 antibody drug conjugate or a pharmaceutically acceptable salt thereof and / or (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof). For example, the term includes inhibition of activity by at least 5%, 10%, 20%, 30%, 40% or more. Thus, inhibition need not be 100%.

[0084] The term "treating" includes administering the pharmaceutical combination of the present invention to a subject in need thereof for the purpose of curing the disease or having an effect on the regression of the disease or delaying the progression of the disease. When referring to a disease, the term "treating" means alleviating the disease (i.e., slowing or arresting or reducing the development of the disease or at least one of its clinical symptoms), preventing or delaying the onset or development or progression of the disease.

[0085] The term "preventing" includes inhibiting or delaying the onset or frequency of a disease or disorder or its symptoms and generally refers to the administration of a drug before signs or symptoms occur, particularly in an at-risk individual.

[0086] The term "subject" refers to both mammals and non-mammals. Mammals refer to any member of the class mammals, including but not limited to humans; non-human primates; cattle, horses, sheep, pigs, rabbits, dogs, and cats. The term "subject" does not limit the subject to a particular age or sex. In some embodiments, the subject is a human.

[0087] The term "adverse event" (AE) is any unfavorable and generally unexpected or unwanted sign (including abnormal laboratory findings), symptom, or disease associated with the use of a medical treatment. For example, an adverse event may be associated with activation of the immune system in response to the treatment or expansion of immune system cells (e.g., T cells) in response to the treatment. A medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity.

[0088] The term "overall survival" or "OS" is the time from the first dose of the study drug to the patient's death from any cause.

[0089] The term "progression-free survival" or "PFS" is the time from the first dose of the study drug to the time of disease progression or death from any cause.

[0090] All numerical ranges herein should be understood to disclose every value and subset of values ​​within the range, regardless of whether they are specifically disclosed otherwise. For example, when any numerical range is mentioned, it should be considered to mention every value within the numerical range, such as every integer within the numerical range. The present invention relates to all values ​​falling within these ranges, all smaller ranges, and the upper or lower limits of the numerical ranges.

[0091] Technical and scientific terms used herein without specific definition have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0092] II. Pharmaceutical Combinations of the Present Invention

[0093] The pharmaceutical combination of the present invention comprises (i) an anti-nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof; and (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0094] (i) Anti-Nectin4 Antibody Drug Conjugate or Pharmaceutically Acceptable Salt thereof

[0095] Antibody-drug conjugates (ADCs) are a technology that uses the specific recognition ability of antibodies for specific antigens on the surface of tumor cells to accurately deliver anti-tumor drugs (such as cytotoxic agents, cytostatics, small molecule chemotherapeutics, etc.) to tumor target cells, causing them to accumulate and release intracellularly, thereby precisely killing tumors. Antibody-drug conjugates generally consist of three parts: an antibody or antibody-like ligand, a small molecule drug, and a linker (connector) that couples the antibody or antibody-like ligand to the drug. Due to their appropriate molecular weight, high stability, strong targeting, and minimal toxic side effects, antibody-drug conjugates have been considered the most promising anti-tumor drugs.

[0096] The anti-nectin 4 antibody-drug conjugate in the drug combination of the present invention consists of three parts: an antibody or an antigen-binding fragment thereof that specifically binds to nectin-4, a small molecule drug, and a linker (connector) that couples the antibody and the small molecule drug.

[0097] In some embodiments, the anti-nectin4 antibody drug conjugate has the molecular formula Ab-[L-CTD]m, wherein

[0098] Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof,

[0099] L represents a connector,

[0100] CTD means drug,

[0101] m represents the average number of drug attachments per Ab molecule.

[0102] In some embodiments, Ab in the molecular formula Ab-[L-CTD]m comprises a heavy chain variable region and a light chain variable region, and the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 1, HCDR2 shown in SEQ ID NO: 2, and HCDR3 shown in SEQ ID NO: 3; and the light chain variable region comprises LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6;

[0103] Preferably, the Ab comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the sequence shown in SEQ ID NO: 7, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereof, and the light chain variable region comprises the sequence shown in SEQ ID NO: 8, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereof;

[0104] For example, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7 or SEQ ID NO: 9, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8 or SEQ ID NO: 10;

[0105] More preferably, the Ab comprises:

[0106] - the heavy chain variable region amino acid sequence shown in SEQ ID NO: 7; and the light chain variable region amino acid sequence shown in SEQ ID NO: 8; or

[0107] - the heavy chain variable region amino acid sequence shown in SEQ ID NO: 9; and the light chain variable region amino acid sequence shown in SEQ ID NO: 10;

[0108] Preferably, the Ab is an IgG antibody; more preferably, it is a human IgG antibody; most preferably, it is a human IgG1 or human IgG4 antibody; wherein the antigen-binding fragment is Fab, Fab', F(ab')2, Fv, single-chain Fv, single-chain Fab.

[0109] In some embodiments, L in the molecular formula Ab-[L-CTD]m is any chemical moiety capable of connecting CTD to Ab via a covalent bond. A cross-linking reagent is a bifunctional or multifunctional reagent that can be used to connect CTD and Ab to form an antibody drug conjugate. Antibody drug conjugates can be prepared using a cross-linking reagent having a reactive functional group that can bind to both CTD and Ab. For example, cysteine, thiol or amine, such as the N-terminus of an antibody or an amino acid side chain such as lysine, can form a bond with the functional group of the cross-linking reagent. Alternatively, the antibody drug conjugate can be prepared by pre-forming a drug-containing linker (i.e., a linker-drug conjugate) and reacting the drug-containing linker with the antibody. In one embodiment, L is a cleavable linker. In another embodiment, L is a non-cleavable linker. In some embodiments, L is an acid-labile linker, a photolabile linker, a peptidase-cleavable linker, an esterase-cleavable linker, a disulfide-cleavable linker, a hydrophilic linker, a precharged linker, a glycosidase-cleavable linker, a phosphodiesterase-cleavable linker, a phosphatase-cleavable linker, or a dicarboxylic acid-based linker.

[0110] In some embodiments, the CTD in the molecular formula Ab-[L-CTD]m is a cytotoxic drug. In preferred embodiments, the CTD is one or more selected from the following: microtubule inhibitors MMAE, DM1, DM4, Tublysin, amanitin, calicheamicin, eribulin, and derivatives thereof; topoisomerase inhibitors SN38, exitecan, and derivatives thereof; and DNA binders PBD, doxorubicin, and derivatives thereof. m is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, even more preferably 3.8-4.2, still more preferably 3.9-4.1, and particularly preferably 4.0.

[0111] [Corrected 08.04.2025 according to Rule 26] In some embodiments, the anti-nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof has the structure shown in Formula Ia and / or Ib below:

[0112] wherein: Ab is an anti-nectin-4 antibody or an antigen-binding fragment thereof as defined above;

[0113] Ar' is any one selected from the following: a substituted or unsubstituted C6-C10 arylene group and a substituted or unsubstituted 5-12 membered heteroarylene group, wherein the substitution refers to the replacement of hydrogen atoms on the group by one or more substituents, and the substituents are selected from any one of the following: halogen (F, Cl, Br or I), haloalkyl (e.g., haloC1-C6 alkyl, preferably haloC1-C4 alkyl, such as trifluoromethyl) and alkoxy (e.g., C1-C6 alkoxy, preferably C1-C4 alkoxy, such as methoxy);

[0114] L1 is -O(CH2CH2O)n- connected to the Ar' group, wherein n is selected from any integer in the range of 1-24, preferably any integer in the range of 1-10, more preferably any integer in the range of 3-5;

[0115] L2 is an enzymatically cleavable fragment, such as a dipeptide, tripeptide, or tetrapeptide, or a combination thereof with a self-releasing structural fragment (i.e., a polypeptide fragment consisting of 2-4 amino acids, or a combination thereof with a self-releasing structural fragment), such as Val-Ala, Val-Ala-PAB, Val-Cit, Val-Cit-PAB, Phe-Lys-PAB, Ala-Ala-Ala, Gly-Gly-Phe-Gly (GGFG), and MAC glucuronide phenol.

[0116] In some embodiments, L2-CTD is VcMMAE, GGFG-Dxd or VC-seco-DUBA; preferably, when Ar' is a substituted or unsubstituted 5-12 membered heteroarylene group, the heteroatom is N; preferably, Ar' is a substituted or unsubstituted C6 arylene group or a substituted or unsubstituted 6 membered heteroarylene group; more preferably, the antibody drug conjugate or a pharmaceutically acceptable salt thereof has the structure of the above-mentioned conjugate ADC-1, conjugate ADC-2, conjugate ADC-3, conjugate ADC-4, conjugate ADC-5, conjugate ADC-6, or conjugate ADC-7.

[0117] In some embodiments, the anti-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof is represented by the following formula:

[0118] Wherein, the anti-nectin-4 antibody is as described above,

[0119] N represents the average number of drug linkages per anti-nectin-4 antibody molecule, and is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, further preferably 3.8-4.2, further preferably 3.9-4.1, and particularly preferably 4.0;

[0120] In another embodiment, the anti-Nectin4 antibody drug conjugate is PADCEV TM(Also known as enfortumab vedotin-ejfv; Chinese name: Vitin-enfortumab), which targets the Nectin-4 protein that is highly expressed in urothelial carcinoma, has been approved by the FDA for the treatment of locally advanced or metastatic urothelial carcinoma. This ADC drug is composed of the human IgG1 monoclonal antibody enfortumab targeting the Nectin-4 protein coupled with the cytotoxic agent MMAE (monomethyl auristatin E, a microtubule disruptor) (Pia M. Challita-Eid et al. (2016) Cancer Res. 76(10):3003-13). However, PADCEVTM can cause serious skin adverse reactions, including Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) (2019 FDA announcement).

[0121] (ii) anti-PD-1 antibodies or antigen-binding fragments thereof

[0122] The anti-PD-1 antibody or antigen-binding fragment thereof in the drug combination of the present invention binds to PD-1 expressed on the surface of T lymphocytes, thereby blocking the binding of PD-1 to its ligand, promoting T lymphocyte activation, proliferation and production of immune-activating cytokines such as IL-2, thereby exerting an anti-tumor effect.

[0123] In one embodiment, the anti-PD-1 antibody in the pharmaceutical combination of the present invention comprises 6 CDRs, wherein HCDR1, HCDR2, and HCDR3 consist of the amino acid sequences shown in SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, respectively, and wherein LCDR1, LCDR2, and LCDR3 consist of the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively. The CDR sequences are CDR sequences defined according to the Kabat numbering scheme.

[0124] Preferably, the anti-PD-1 antibody comprises a heavy chain variable region VH and a light chain variable region VL, wherein the heavy chain variable region comprises the sequence of SEQ ID NO: 17 or a sequence having at least 90%, 95%, 98% or 99% identity thereto, and the light chain variable region comprises the sequence of SEQ ID NO: 18 or a sequence having at least 90%, 95%, 98% or 99% identity thereto.

[0125] Preferably, the anti-PD-1 antibody comprises SEQ ID NO: 19 or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO: 20 or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto.

[0126] Preferably, the anti-PD-1 antibody is toripalimab.

[0127] In another embodiment, the anti-PD-1 antibody in the pharmaceutical combination of the present invention comprises 6 CDRs, wherein HCDR1, HCDR2, and HCDR3 consist of the amino acid sequences shown in SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23, respectively, and wherein LCDR1, LCDR2, and LCDR3 consist of the amino acid sequences shown in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, respectively. The CDR sequences are defined according to the Kabat numbering scheme.

[0128] Preferably, the anti-PD-1 antibody comprises a heavy chain variable region VH and a light chain variable region VL, wherein the heavy chain variable region comprises the sequence of SEQ ID NO: 27 or a sequence having at least 90%, 95%, 98% or 99% identity thereto, and the light chain variable region comprises the sequence of SEQ ID NO: 28 or a sequence having at least 90%, 95%, 98% or 99% identity thereto.

[0129] Preferably, the anti-PD-1 antibody comprises SEQ ID NO: 29 or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO: 30 or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto.

[0130] Preferably, the anti-PD-1 antibody is pembrolizumab.

[0131] Other anti-PD-1 antibodies contemplated for use in the drug combinations of the present invention include MEDI0680 (U.S. Patent No. 8,609,089), BGB-A317 (U.S. Patent Publication No. 2015 / 0079109), INCSHR1210 (SHR-1210) (WO2015 / 085847), REGN-2810 (WO2015 / 112800), PDR001 (WO2015 / 112900), TSR-042 (ANB011) (WO2014 / 179664), and STI-1110 (WO2014 / 194302).

[0132] III. Uses of the pharmaceutical combination of the present invention and methods of treatment using the pharmaceutical combination of the present invention

[0133] The present invention provides the aforementioned pharmaceutical combination of the present invention for use in preventing and / or treating the severity of at least one symptom or indication of cancer in an individual or inhibiting the growth of cancer cells.

[0134] The present invention provides a method for preventing or treating cancer, comprising administering an effective amount of the pharmaceutical combination of the present invention to an individual in need thereof. The effective amount includes a preventive effective amount and a therapeutic effective amount.

[0135] The present invention provides use of the aforementioned pharmaceutical combination of the present invention in preparing a medicament for preventing or treating cancer.

[0136] Cancers of the present invention include solid tumors and hematological malignancies, such as urothelial carcinoma, bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, gastric cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute lymphocytic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, prostate cancer, colorectal cancer, colon cancer, glioma, mesothelioma. The cancer is preferably advanced cancer, refractory cancer and / or cancer resistant to chemotherapy, more preferably advanced solid tumors, (confirmed by histology or cytology) cannot be surgically removed or metastatic advanced solid tumors. The cancer is preferably intestinal cancer (including colorectal cancer, colon cancer), lung cancer (including non-small cell lung cancer). The cancer is preferably advanced recurrent or metastatic cancer (advanced malignant tumor).

[0137] The pharmaceutical combinations of the present invention may be administered to individuals who have been treated with one or more prior therapies but who have subsequently relapsed or metastasized.

[0138] The pharmaceutical combination of the present invention is administered to an individual who exhibits elevated levels of one or more cancer-associated biomarkers [e.g., nectin-4, programmed death ligand 1 (PD-L1)]. For example, a prophylactically effective amount or a therapeutically effective amount of the pharmaceutical combination of the present invention is administered to an individual who exhibits elevated levels of nectin-4 and / or elevated levels of PD-L1.

[0139] The anti-nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof in the pharmaceutical combination of the present invention can be administered to an individual in need thereof in one or more doses. For example, when multiple doses are administered, the next dose is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the previous dose.

[0140] The anti-PD-1 antibody or antigen-binding fragment thereof in the pharmaceutical combination of the present invention can be administered to a subject in need thereof in one or more doses. For example, in the case of administering multiple doses, the next dose is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the previous dose.

[0141] The pharmaceutical combination of the present invention can be in any dosage form known to those skilled in the art, such as tablets, capsules, granules, syrups, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, creams, and injections. The anti-nectin 4 antibody-drug conjugate or its pharmaceutically acceptable salt and the anti-PD-1 antibody or its antigen-binding fragment can each be in a separate dosage form, which can be different or the same. The anti-nectin 4 antibody-drug conjugate or its pharmaceutically acceptable salt can be in an intravenous dosage form or an intraperitoneal injection. The anti-PD-1 antibody or its antigen-binding fragment can be in an intravenous dosage form or an intraperitoneal injection.

[0142] The pharmaceutical combination of the present invention may be a pharmaceutical dosage unit, such as a single pharmaceutical dosage unit.

[0143] The anti-nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof and the anti-PD-1 antibody or an antigen-binding fragment thereof in the pharmaceutical combination of the present invention can be administered separately, simultaneously or sequentially.

[0144] In some embodiments, administration of at least one cycle of the drug combination of the present invention results in an increase, preferably a synergistic increase, in the patient's progression-free survival (PFS) or overall survival (OS), compared to patients administered an anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof as a monotherapy or with patients administered an anti-PD-1 antibody or its antigen-binding fragment as a monotherapy. In some embodiments, administration of at least one cycle of the drug combination of the present invention results in an increase in the patient's PFS by at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 2 years or more, compared to patients administered an anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof as a monotherapy or with patients administered an anti-PD-1 antibody or its antigen-binding fragment as a monotherapy. In some embodiments, administration of at least one cycle of the drug combination of the present invention results in an increase in the patient's OS by at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 2 years, or longer, compared to patients administered monotherapy with an anti-nectin 4 antibody drug conjugate, or a pharmaceutically acceptable salt thereof, or with monotherapy with an anti-PD-1 antibody, or an antigen-binding fragment thereof.

[0145] Compared to monotherapy with anti-Nectin4 antibody drug conjugates or pharmaceutically acceptable salts thereof or monotherapy with anti-PD-1 antibodies or antigen-binding fragments thereof, the drug combination of the present invention results in an increase, preferably a synergistic increase, in the inhibition of tumor growth. In some embodiments, compared to monotherapy with anti-Nectin4 antibody drug conjugates or pharmaceutically acceptable salts thereof or monotherapy with anti-PD-1 antibodies or antigen-binding fragments thereof, the drug combination of the present invention results in tumor growth being inhibited by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70% or about 80%. In some embodiments, administration of the drug combination of the present invention results in increased tumor regression, tumor shrinkage and / or disappearance. In some embodiments, the drug combination of the present invention prevents tumor recurrence and / or increases survival duration in individuals compared to untreated individuals or monotherapy with administration of an anti-Nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof or monotherapy with administration of an anti-PD-1 antibody or its antigen-binding fragment, for example, by increasing survival duration by more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 12 months, more than 18 months, more than 24 months, more than 36 months, or more than 48 months. In some embodiments, the drug combination of the present invention can increase progression-free survival or overall survival.

[0146] In some embodiments, administration of the drug combination of the present invention to an individual with cancer results in complete disappearance of the tumor (a "complete response"). In some embodiments, administration of the drug combination of the present invention to an individual with cancer results in a reduction of tumor cells or tumor size by at least 30% or more (a "partial response"). The reduction of the tumor can be measured by any method known in the art, such as X-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0147] In some embodiments, the drug combination of the present invention can reduce adverse events caused by the administration of anti-PD-1 antibodies or antigen-binding fragments thereof and / or anti-nectin4 antibody drug conjugates or pharmaceutically acceptable salts thereof, such as hematological toxic reactions, non-hematological toxic reactions or other toxic reactions, such as pneumonia, diarrhea, renal insufficiency, rash, endocrine diseases, etc.

[0148] IV. Kits of the Present Invention

[0149] Another object of the present invention is to provide a kit comprising the pharmaceutical combination of the present invention, preferably in the form of a pharmaceutical dosage unit, whereby dosage units can be provided according to a dosing regimen or a drug administration interval.

[0150] In one embodiment, the kit of parts of the present invention comprises in the same package:

[0151] - a first container containing a pharmaceutical composition for administration, wherein the pharmaceutical composition comprises an anti-nectin 4 antibody drug conjugate or a pharmaceutically acceptable salt thereof;

[0152] - A second container containing a pharmaceutical composition for administration, the pharmaceutical composition comprising an anti-PD-1 antibody or antigen-binding fragment thereof.

[0153] The various embodiments / technical solutions described in the present invention and the features in each embodiment / technical solution should be understood to be able to be combined with each other arbitrarily, and the various solutions obtained by these combinations are included in the scope of the present invention, just as the solutions obtained by these combinations are specifically and one by one listed in this document, unless the context clearly shows otherwise.

[0154] Example

[0155] The following examples are presented to provide those of ordinary skill in the art with a complete disclosure and description of how to make the compositions of the present invention and methods of use thereof and are not intended to limit the scope of what the inventors regard as their invention.

[0156] Example 1 Preparation of anti-Nectin4 antibody-drug conjugate

[0157] According to the method described in Chinese patent application CN202210475286.3, hH2L1 (an anti-nectin4 antibody) and a drug-containing linker (i.e., a linker-drug conjugate) were used to prepare the anti-nectin4 antibody drug conjugate (also referred to as "anti-nectin4-ADC") shown below. The DAR was calculated by hydrophobic interaction chromatography (HIC): N = 4.0

[0158] Example 2: In vivo efficacy of anti-nectin4-ADC combined with anti-PD-1 antibody in tumor-bearing mice

[0159] In this example, PD-1 tumor-bearing HuGEMM-C57BL / 6J mice were used as test animals to evaluate and compare the anti-tumor effects of anti-Nectin4-ADC alone, anti-PD-1 antibody alone, or anti-Nectin4-ADC combined with anti-PD-1 antibody on subcutaneous xenograft tumors in mice.

[0160] 2.1 Test drugs and materials

[0161] 2.1.1 Test drugs

[0162] (Lot number: W004692, purchased from MSD Ireland (Carlow)), which is the anti-PD-1 antibody pembrolizumab injection;

[0163] Anti-Nectin4-ADC: prepared according to Example 1.

[0164] 2.1.2 Test animals

[0165] Mice: SPF (Specific Pathogen Free) grade 6-8 week old female PD-1 HuGEMM-C57BL / 6J mice (purchased from Shanghai Dishi Biotechnology Co., Ltd.)

[0166] 2.2 Experimental methods

[0167] Mouse colon cancer cells MC38-hNECTIN4 were used to inoculate PD-1HuGEMM-C57BL / 6J mice, 6-8 weeks old, female (purchased from Shanghai Dishi Biotechnology Co., Ltd.), to prepare tumor-bearing PD-1HuGEMM-C57BL / 6J mice.

[0168] Specifically, a tumor-bearing mouse model was established using mouse colon cancer cells MC38-hNECTIN4 (purchased from Shanghai South Model Organisms Technology Co., Ltd.). MC38-hNECTIN4 cells were cultured in DMEM / High medium (containing 10% inactivated fetal bovine serum) (in an incubator at 37°C and 5% CO2). MC38-hNECTIN4 cells in the logarithmic growth phase were taken and resuspended in PBS to a suitable concentration for subcutaneous tumor inoculation in mice; the cell suspension was inoculated subcutaneously in the right hindquarters of PD-1HuGEMM-C57BL / 6J mice using a 1 mL syringe, 100 μL / mouse, and each mouse was inoculated with approximately 5.0×10 6 The PD-1 tumor-bearing HuGEMM-C57BL / 6J mouse model was established by inoculating MC38-hNECTIN4 tumor cells.

[0169] This experiment used anti-Nectin4-ADC (prepared according to Example 1) and anti-PD-1 antibody (Batch number: W004692, purchased from MSD Ireland (Carlow)) for group comparison studies (see Table 1). Grouping was defined as day 0, and mice were given the first injection of 10 μL / g of the drug on the day of grouping; the solvent group was given the same volume of solvent (normal saline); the specific dosage and dosing schedule are shown in Table 1. Tumor volume was measured twice a week, and the mice were weighed and the data were recorded. The experiment was completed, the experimental endpoint was reached, or the tumor volume reached 3000 mm 3The animals were killed by CO2 anesthesia, and the tumors were removed by dissection and photographed.

[0170] Table 1. Dosage and administration schedule

[0171] 2.3 Experimental Results and Conclusions

[0172] The results are shown in Figure 1 and Table 2. There were no significant differences in the single-drug groups compared to the saline control group. However, the two combination treatment groups, the anti-Nectin4-ADC 2 mg / kg + Keytruda 3 mg / kg group and the anti-Nectin4-ADC 5 mg / kg + Keytruda 3 mg / kg group, showed significant differences compared to the saline control group (p<0.05 and p<0.001, respectively).

[0173] The anti-tumor effect of the anti-Nectin4-ADC 2mg / kg + Keytruda 3mg / kg combination group was better than that of the Keytruda 3mg / kg single-drug group, and there was also a significant difference compared with the anti-Nectin4-ADC 5mg / kg single-drug group and the anti-Nectin4-ADC 10mg / kg single-drug group (p2 value <0.01).

[0174] The anti-nectin4-ADC 5 mg / kg + Keytruda 3 mg / kg combination demonstrated superior anti-tumor efficacy compared to both the anti-nectin4-ADC 10 mg / kg and Keytruda 3 mg / kg monotherapy groups, with significant differences compared to either monotherapy group (p<0.01). Although the total dose of the combination (anti-nectin4-ADC 5 mg / kg + Keytruda 3 mg / kg) was lower than that of the monotherapy group (anti-nectin4-ADC 10 mg / kg), it demonstrated significantly superior efficacy compared to the monotherapy groups. This suggests that the combination of anti-nectin4-ADC and Keytruda demonstrates a significant synergistic therapeutic effect.

[0175] In this experiment, mice in the saline control group, anti-Nectin4-ADC 5 and 10 mg / kg, Keytruda 3 mg / kg, anti-Nectin4-ADC 2 mg / kg + Keytruda 3 mg / kg, and anti-Nectin4-ADC 5 mg / kg + Keytruda 3 mg / kg groups showed no significant weight loss (Figure 2). One mouse in Group 2 and Group 4 died due to tumor rupture on day 19 and day 15, respectively.

[0176] Table 2. Pharmacological efficacy analysis of MC38-hNectin4 in subcutaneous mouse colon cancer model according to tumor volume of each group of mice Note: a. Tumor volume data are expressed as “mean ± standard error” (Mean ± SEM);

[0177] bT / C%=T / C×100%; TGI%=(1-T / C)×100%, where T and C are the absolute tumor volumes of each model treatment group and the saline control group on the 19th day of the experiment (Vt-V0).

[0178] Example 3 In vivo efficacy of anti-Nectin4-ADC combined with anti-PD-1 antibody in tumor-bearing mice (Part 2)

[0179] In this example, PD-1-HU tumor-bearing mice were used as test animals to evaluate and compare the anti-tumor effects of anti-Nectin4-ADC alone, anti-PD-1 antibody alone, or anti-Nectin4-ADC combined with anti-PD-1 antibody on subcutaneous xenograft tumors in mice.

[0180] 3.1 Test drugs and materials

[0181] 3.1.1 Test drugs

[0182] (Purchased from Junshi Biosciences), the main ingredient of this product is Toripalimab, an anti-PD-1 antibody.

[0183] Anti-Nectin4-ADC: prepared according to Example 1.

[0184] (Enfortumab Vedotin): White lyophilized powder, 30 mg / vial, batch number 102374, store at 2-8°C in the dark. It is an ADC drug that is a nectin-4 targeting antibody and microtubule inhibitor.

[0185] 3.1.2 Test animals

[0186] Mice: SPF-grade 6-week-old female PD-1-HU mice (purchased from Shanghai Nanmo Biotechnology Co., Ltd.).

[0187] 3.2 Experimental methods

[0188] PD-1-HU mice, 6 weeks old, female (purchased from Shanghai Nanmo Biotechnology Co., Ltd.), were inoculated with mouse colon cancer cells MC38 / Nectin4 hPD-L1 to prepare tumor-bearing PD-1-HU mice.

[0189] Specifically, a tumor-bearing mouse model was established using mouse colon cancer cells MC38 / Nectin4 hPD-L1 (Shanghai South Model Organisms Science Co., Ltd.). MC38 / Nectin4hPD-L1 cells were cultured in DMEM / High medium (containing 10% inactivated fetal bovine serum) (in an incubator at 37°C and 5% CO2). MC38 / Nectin4 hPD-L1 cells in the logarithmic growth phase were taken and resuspended in PBS to a suitable concentration for subcutaneous tumor inoculation in PD-1-HU mice; the cell suspension was inoculated into the right hindquarters of PD-1-HU mice using a 1 mL syringe, 100 μL / mouse, and each mouse was inoculated with approximately 5.0×10 5 MC38 / Nectin4 hPD-L1 tumor cells were used to establish a tumor-bearing PD-1-HU mouse model.

[0190] This study used anti-Nectin4-ADC and anti-PD-1 antibody Tuoyi (purchased from Junshi Biosciences) for group comparison (see Table 3). 3 Afterwards, the animals were randomly divided into groups according to tumor volume, with the day of grouping designated as day 0. The solvent group was given the same volume of solvent (normal saline); the specific dosage and dosing schedule are shown in Table 3. Tumor volume was measured twice a week, and the mice were weighed and the data were recorded. The experiment was completed, the experimental endpoint was reached, or the tumor volume reached 3000 mm 3 The animals were killed by CO2 anesthesia, and the tumors were removed by dissection and photographed.

[0191] Table 3. Dosage and administration schedule Note: IP-intraperitoneal injection, administration started on D0; BIW: twice a week.

[0192] The results are shown in Figures 3 and 4 and Table 4. Compared with the vehicle group, the anti-nectin4-ADC 3 mg / kg monotherapy group, the Tuoyi 5 mg / kg monotherapy group, and the two combination therapy groups (anti-nectin4-ADC 3 mg / kg + Tuoyi 5 mg / kg group and PADCEV 3 mg / kg + Tuoyi 5 mg / kg group) all showed some efficacy against MC38 / Nectin4 hPD-L1 subcutaneous xenografts. Compared with the Tuoyi monotherapy group, the two combination therapy groups (anti-nectin4-ADC 3 mg / kg + Tuoyi 5 mg / kg group and PADCEV 3 mg / kg + Tuoyi 5 mg / kg group) showed superior anti-tumor efficacy. The anti-nectin4-ADC 3 mg / kg + Tuoyi 5 mg / kg combination therapy group showed superior anti-tumor efficacy compared with the PADCEV 3 mg / kg + Tuoyi 5 mg / kg group. The anti-nectin4-ADC 3mg / kg plus TuoYi 5mg / kg group showed superior anti-tumor efficacy compared to the anti-nectin4-ADC 3mg / kg monotherapy group. Tumor-bearing mice tolerated all of these drugs well, with no significant weight loss or other symptoms, and no animals died during the experiment.

[0193] Table 4. Efficacy against MC38 / Nectin4 hPD-L1 subcutaneous xenografts in mice Note: TV: tumor volume, mm 3 .

[0194] Example 4 Treatment of urothelial carcinoma with anti-Nectin4-ADC combined with anti-PD-1 antibody

[0195] This Phase Ib / II, open-label, single-arm, multicenter study in patients with locally advanced or metastatic urothelial carcinoma consists of a screening phase, a treatment phase of up to 2 years, and a safety / efficacy follow-up phase.

[0196] 1. Clinical research objectives:

[0197] The primary objective is to evaluate the safety and tolerability of an anti-nectin4-ADC combined with toripalimab in patients with locally advanced or metastatic urothelial carcinoma. Secondary objectives include the following:

[0198] 1) Evaluate the preliminary efficacy of an anti-nectin4 ADC combined with toripalimab in patients with locally advanced or metastatic urothelial carcinoma;

[0199] 2) Evaluate the pharmacokinetic (PK) characteristics of anti-nectin4-ADC;

[0200] 3) evaluate the immunogenicity of anti-Nectin4-ADC;

[0201] 4) Evaluate the relationship between the biomarkers Nectin-4 and PD-L1 in tumor tissue and efficacy or prognosis.

[0202] The study protocol and all amendments were approved by the institutional ethics committees of all participating centers.

[0203] 2. Test drug:

[0204] The active components of the test drug described in this example are: anti-Nectin4-ADC and anti-PD-1 antibody.

[0205] Anti-Nectin4-ADC: prepared according to Example 1, with a specification of 20 mg / bottle, is a conjugate of anti-Nectin-4 antibody and MMAE;

[0206] The anti-PD-1 antibody is toripalimab injection, manufactured by Shanghai Junshi Biotechnology Co., Ltd.

[0207] 3. Patient inclusion criteria

[0208] 1) All subjects or guardians must personally sign a written informed consent form approved by the ethics committee before commencing any screening procedure;

[0209] 2) Age between 18 and 80 years old (inclusive) when signing the informed consent form, regardless of gender;

[0210] 3) Eastern Cooperative Oncology Group (ECOG) score of 0–1;

[0211] 4) Patients with locally advanced or metastatic urothelial carcinoma (including bladder, ureter, renal pelvis and urethra) confirmed by histopathology, with squamous differentiation, glandular differentiation or mixed types of urothelial carcinoma also meet the requirements and cannot undergo radical surgical resection;

[0212] 5) Previous treatment for locally advanced or metastatic urothelial carcinoma: failure of previous standard treatment or no previous systemic treatment;

[0213] 6) Must provide archived tumor tissue samples or undergo a new tumor tissue biopsy;

[0214] 7) The expected survival period is not less than 12 weeks;

[0215] 8) According to the RECIST v1.1 criteria, at least one measurable lesion was present during the screening period; for patients with a clear history of radiotherapy, the measurable lesion showed clear progression outside the radiation field or after completion of radiotherapy;

[0216] 9) The subject's organ function must meet the following criteria. If the subject has recently received a blood transfusion or growth factor therapy, a hematological examination must be performed ≥7 days after any growth factor therapy or ≥14 days after any blood transfusion:

[0217] Absolute neutrophil count (ANC) ≥1.5 × 10 9 / L;

[0218] Platelet count (PLT) ≥100 × 10 9 / L;

[0219] Hemoglobin (Hb) ≥ 90 g / L;

[0220] Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ≤ 3 times the upper limit of normal (ULN);

[0221] Total bilirubin ≤ 1.5 times ULN (total bilirubin should be < 3 times ULN in subjects with Gilbert's syndrome);

[0222] International Normalized Ratio (INR) ≤ 1.5 times ULN;

[0223] Creatinine clearance ≥ 50 mL / min (calculated according to the Cockcroft-Gault formula), or serum creatinine ≤ 1.5 times the ULN.

[0224] 10) Male or female subjects must meet any of the following conditions:

[0225] Women of infertile potential, specifically those who have undergone hysterectomy, bilateral oophorectomy, or bilateral tubal ligation or are postmenopausal;

[0226] Women of childbearing age with a negative serum pregnancy test within 7 days before the first dose should use effective contraceptive measures throughout the study and for 180 days after the end of the treatment, such as double-barrier contraceptive methods, condoms, oral or injectable contraceptives, and intrauterine devices;

[0227] Male subjects who have undergone vasectomy or agree to use contraception during the study treatment phase and within 180 days of the last dose of study drug.

[0228] 11) Able to understand and perform planned visits, treatments, laboratory tests, and other study procedures.

[0229] Exclusion criteria:

[0230] 1) Received chemotherapy, radiotherapy or other anti-tumor treatment within 21 days before the first study drug treatment;

[0231] 2) Patients who have undergone major surgery within 28 days before the first study drug treatment (defined as requiring general anesthesia and hospitalization for >24 hours), except for minor surgeries (such as tooth extraction, puncture, etc.) that the investigator judges will not affect participation in the trial;

[0232] 3) (Cohort expansion phase only) Previous use of PD-1, PD-L1, or PD-L2 inhibitors for locally advanced or metastatic urothelial carcinoma, including but not limited to pembrolizumab, atezolizumab, and tislelizumab;

[0233] 4) Patients who have received MMAE-conjugated antibody-drug conjugate therapy before, such as Enfortumab vedotin;

[0234] 5) The subject continues to have clinically significant toxicity (Grade 2 or above, except for alopecia and pigmentation) related to previous treatment (including systemic therapy, radiotherapy or surgery);

[0235] 6) Subjects have grade 2 or higher peripheral neuropathy;

[0236] 7) Subjects with poor blood sugar control;

[0237] 8) Investigators assess subjects for conditions that may increase the risk of corneal disease before the first dose;

[0238] 9) Subjects with clinically significant cardiovascular and cerebrovascular diseases within 6 months before the first study drug use, including but not limited to: ① acute myocardial infarction; ② unstable angina; ③ cerebrovascular accident (excluding lacunar infarction that is asymptomatic and does not require clinical intervention); ④ history of clinically significant ventricular arrhythmias (such as sustained ventricular tachycardia, ventricular fibrillation, torsades de pointes); ⑤ New York Heart Association (NYHA) class III or IV congestive heart failure; ⑥ QTcF ≥ 470ms (female) or ≥ 450ms (male) or history or family history of congenital long QT syndrome; ⑦ baseline left ventricular ejection fraction (LVEF) < 50% or severe ventricular wall motion disorder detected by echocardiography (ECHO); ⑧ subjects with hypertension poorly controlled by drug therapy; ⑨ other arrhythmias determined by the investigator to be unsuitable for inclusion in this study;

[0239] 10) Active infection, including but not limited to: hepatitis B virus infection (HBsAg positive and HBV-DNA>500IU / mL), hepatitis C virus infection (HCV-Ab positive and HCV-RNA positive), human immunodeficiency virus infection (HIV-Ab positive), active Mycobacterium tuberculosis infection, or other active infection requiring systemic treatment within 14 days before the first study drug (excluding routine preventive anti-infection treatment);

[0240] 11) Subjects have other severe or uncontrolled diseases that the investigator considers unsuitable for participation in this clinical trial or that may affect the subject's compliance with the study protocol, including but not limited to: ① Severe respiratory diseases, such as severe interstitial lung disease, severe asthma, etc.; ② Severe arteriovenous thrombotic events, such as pulmonary embolism, deep vein thrombosis (except for asymptomatic intermuscular vein thrombosis that does not require special treatment); ③ Large amounts of pleural effusion or ascites accompanied by clinical symptoms or requiring symptomatic treatment;

[0241] 12) Subjects with central nervous system (CNS) metastases and / or carcinomatous meningitis. Subjects who have received treatment for brain metastases may be considered for participation in this study, provided that their condition has been stable for at least 3 months, no disease progression has occurred as determined by imaging examination within 4 weeks before the first dose of the study, all neurological symptoms have fully recovered, there is no evidence of new or enlarged brain metastases, and radiation, surgery, or glucocorticoid treatment has been stopped for at least 28 days before the first dose of the study treatment. Carcinomatous meningitis should be excluded regardless of whether their clinical condition is stable;

[0242] 13) The subject has previously received allogeneic hematopoietic stem cell transplantation or solid organ transplantation;

[0243] 14) The subject has a history of drug abuse or suffers from mental illness, which may affect the trial compliance;

[0244] 15) Patients with known allergy or intolerance to the study drug or any of its components;

[0245] 16) Subjects who have used P-glycoprotein (P-gp) inhibitors or inducers, or strong CYP3A4 inhibitors or inducers within 14 days before the first study medication;

[0246] 17) Subjects who have used high-dose glucocorticoids (>10 mg / day prednisone or equivalent dose of hormones) or other immunosuppressive drugs within 14 days before the first study medication;

[0247] 18) The subject has received a live vaccine within 28 days before the first study medication or plans to receive any live vaccine during the study;

[0248] 19) The subject has received / used other clinical research drugs or experimental medical devices within 28 days before the first study medication;

[0249] 20) Active autoimmune disease requiring systemic treatment within 2 years before the first study medication;

[0250] 21) Subjects who have had other malignant tumors within 3 years before the first study drug use, excluding cancers that have been radically treated and are expected to be cured, such as basal skin cancer or squamous cell skin cancer, localized low-risk prostate cancer, papillary thyroid carcinoma, or any type of carcinoma in situ that has been radically resected, such as cervical carcinoma in situ, breast ductal carcinoma in situ, etc.;

[0251] 22) Other circumstances that the researcher judges to be unsuitable for inclusion in this study.

[0252] Patients who met the above criteria were enrolled and received anti-Nectin4-ADC combined with toripalimab treatment.

[0253] 4. Clinical dosing regimen

[0254] Enrolled subjects received anti-Nectin4-ADC via intravenous infusion on day 1 (D1) and day 8 (D8) of each cycle, and toripalimab via intravenous infusion on D1 of each cycle, with each cycle consisting of 21 days. The doses of anti-Nectin4-ADC in the dose-finding phase were 1.0 mg / kg and 1.25 mg / kg, respectively, and in the cohort expansion phase was 1.25 mg / kg. The dose of toripalimab was 240 mg. All subjects received anti-Nectin4-ADC combined with toripalimab until disease progression, intolerable toxicity, death, withdrawal of informed consent, loss to follow-up, or completion of 2 years of combined treatment, whichever occurred first.

[0255] 5. Results and Summary

[0256] Antitumor activity

[0257] The following clinical results have been obtained for the combination therapy: A total of 15 subjects completed at least one post-baseline tumor assessment, and all subjects experienced varying degrees of target lesion reduction. All patients with a PR response were treated in the same group. Among these patients, four evaluable patients who had previously received platinum-based chemotherapy and PD-1 / L1 therapy and received the anti-nectin4 ADC (1.25 mg / kg) combination therapy in this study achieved an ORR of 75% (3 / 4).

[0258] In a published study (NCT05216965 / CTR20220106) of anti-Nectin4-ADC monotherapy (1.25 mg / kg) for the treatment of patients with locally advanced or metastatic urothelial carcinoma who had previously received platinum-containing chemotherapy and PD-1 / L1 therapy, 37 patients were evaluable and the ORR was 62% (23 / 37).

[0259] In a study of toripalimab for the treatment of patients with locally advanced or metastatic urothelial carcinoma who had previously received platinum-containing chemotherapy (POLARIS-03, NCT03113266 / CTR20170347), the ORR of 151 patients was 25.8% (39 / 151).

[0260] Safety Assessment

[0261] In the anti-nectin4-ADC (1.25 mg / kg) combination treatment group, common TRAEs (all grades ≥10%) were rash, anemia, increased aspartate aminotransferase, increased blood creatinine, increased alanine aminotransferase, nausea, decreased white blood cell count, hypercholesterolemia, alopecia, decreased neutrophil count, constipation, hypertriglyceridemia, pruritus, decreased platelet count, proteinuria, fatigue, decreased appetite, and decreased weight.

[0262] Only one patient experienced a dose reduction of the anti-nectin4-ADC due to an adverse event. No TRAEs resulted in death. These results suggest that the combination of an anti-nectin4-ADC and an anti-PD-1 antibody has promising anti-urothelial cancer efficacy.

[0263] Although some representative embodiments and details have been shown for the purpose of illustrating the present invention, it will be apparent to those skilled in the art that various changes and modifications can be made thereto without departing from the scope of the subject invention. In this regard, the scope of the present invention is limited only by the following claims.

[0264] Exemplary sequences

Claims

1. Drug combination, including (i) an anti-Nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof; and (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof.

2. The pharmaceutical combination according to claim 1, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises (i) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 11, HCDR2 shown in SEQ ID NO: 12, and HCDR3 shown in SEQ ID NO: 13; and the light chain variable region comprises LCDR1 shown in SEQ ID NO: 14, LCDR2 shown in SEQ ID NO: 15, and LCDR3 shown in SEQ ID NO: 16; or (ii) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1 shown in SEQ ID NO:21, HCDR2 shown in SEQ ID NO:22, and HCDR3 shown in SEQ ID NO:23; and the light chain variable region comprises LCDR1 shown in SEQ ID NO:24, LCDR2 shown in SEQ ID NO:25, and LCDR3 shown in SEQ ID NO:26; Preferably, the anti-PD-1 antibody or antigen-binding fragment thereof comprises (i) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a sequence as set forth in SEQ ID NO:17, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical thereto, and the light chain variable region comprises a sequence as set forth in SEQ ID NO:18, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical thereto; or (ii) a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a sequence as set forth in SEQ ID NO:27, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and the light chain variable region comprises a sequence as set forth in SEQ ID NO:28, or a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto; More preferably, the anti-PD-1 antibody comprises (i) SEQ ID NO:19 or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO:20 or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto; or (ii) SEQ ID NO:29, or a heavy chain sequence at least 90%, 95%, 98% or 99% identical thereto and SEQ ID NO:30, or a light chain sequence at least 90%, 95%, 98% or 99% identical thereto.

3. The drug combination according to claim 1 or 2, wherein the anti-PD-1 antibody or its antigen-binding fragment is an IgG antibody; preferably, it is a human IgG antibody; more preferably, it is a human IgG1 or human IgG4 antibody; wherein the antigen-binding fragment is Fab, Fab', F(ab')2, Fv, single-chain Fv, single-chain Fab.

4. The drug combination according to any one of claims 1 to 3, wherein the anti-nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof has a molecular formula Ab-[L-CTD]m, wherein Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof, L represents a linker, CTD represents a drug, and m represents the average number of drug connections relative to each molecule of Ab; preferably, CTD is a cytotoxic drug, preferably, CTD is one or more selected from the following: microtubule inhibitors MMAE, DM1, DM4, Tublysin, amanita chalcone, calicheamicin, eribulin and derivatives of the drugs; topoisomerase inhibitors SN38, exitecan and derivatives of the drugs; DNA binders PBD, doxorubicin and derivatives of the drugs; preferably, m is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, further preferably 3.8-4.2, further preferably 3.9-4.1, and particularly preferably 4.

0.

5. The drug combination according to claim 4, wherein the anti-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof has a molecular formula Ab-[L-CTD]m, wherein Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof, and the Ab comprises a heavy chain variable region and a light chain variable region, and the heavy chain variable region comprises HCDR1 shown in SEQ ID NO: 1, HCDR2 shown in SEQ ID NO: 2, and HCDR3 shown in SEQ ID NO: 3; and the light chain variable region comprises LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6; Preferably, the Ab comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the sequence shown in SEQ ID NO:7, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereto, and the light chain variable region comprises the sequence shown in SEQ ID NO:8, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity thereto; For example, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:7 or SEQ ID NO:9, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:8 or SEQ ID NO:10; More preferably, the Ab comprises: (i) the heavy chain variable region amino acid sequence shown in SEQ ID NO:7; and the light chain variable region amino acid sequence shown in SEQ ID NO:8; or (ii) the heavy chain variable region amino acid sequence shown in SEQ ID NO:9; and the light chain variable region amino acid sequence shown in SEQ ID NO:10; Preferably, the Ab is an IgG antibody; more preferably, it is a human IgG antibody; most preferably, it is a human IgG1 or human IgG4 antibody; wherein the antigen-binding fragment is Fab, Fab', F(ab')2, Fv, single-chain Fv, single-chain Fab.

6. The pharmaceutical combination according to any one of claims 1 to 5, wherein the anti-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof has a structure as shown in the following formula Ia and / or Ib: in: Ab is an anti-nectin-4 antibody or an antigen-binding fragment thereof; Ar' is any one selected from the following: substituted or unsubstituted C6-C10 arylene and substituted or unsubstituted 5-12 membered heteroarylene, wherein the substitution refers to the replacement of hydrogen atoms on the group by one or more substituents, and the substituents are selected from the following Any of: halogen (F, Cl, Br or I), haloalkyl (e.g. haloC1-C6 alkyl, preferably haloC1-C4 alkyl, such as trifluoromethyl) and alkoxy (e.g. C1-C6 alkoxy, preferably C1-C4 alkoxy, such as methoxy); L1 is -O(CH2CH2O)n- connected to the Ar' group, wherein n is selected from any integer in the range of 1-24, preferably any integer in the range of 1-10, more preferably any integer in the range of 3-5; L2 is an enzyme cleavage fragment, such as a dipeptide, a tripeptide, or a tetrapeptide, or a combination thereof with a self-releasing structural fragment (i.e., a polypeptide fragment consisting of 2-4 amino acids, or a combination thereof with a self-releasing structural fragment), such as Val-Ala, Val-Ala-PAB, Val-Cit, Val-Cit-PAB, Phe-Lys-PAB, Ala-Ala-Ala, Gly-Gly-Phe-Gly (GGFG), and MAC glucuronide phenol.

7. The pharmaceutical combination according to claim 6, wherein L2-CTD is VcMMAE, GGFG-Dxd or VC-seco-DUBA; preferably, when Ar' is a substituted or unsubstituted 5-12-membered heteroarylene group, the heteroatom is N; preferably, Ar' is a substituted or unsubstituted C6 arylene group or a substituted or unsubstituted 6-membered heteroarylene group; more preferably, the antibody drug conjugate or a pharmaceutically acceptable salt thereof has the following structure: Conjugate ADC-1: Conjugate ADC-2: Conjugate ADC-3: Conjugate ADC-4: Conjugate ADC-5: Conjugate ADC-6: Conjugate ADC-7:

8. The pharmaceutical combination according to claim 7, wherein (i) The anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof is represented by the following formula: Wherein, the anti-nectin-4 antibody is as described in claim 5, N represents the average number of drug connections per anti-nectin-4 antibody molecule, which is 1.0-5.0, preferably 3.0-4.2, more preferably 3.5-4.5, further preferably 3.8-4.2, further preferably 3.9-4.1, and particularly preferably 4.0; and (ii) the anti-PD-1 antibody or antigen-binding fragment thereof is an anti-PD-1 antibody or antigen-binding fragment thereof selected from toripalizumab or pembrolizumab.

9. Use of the drug combination according to any one of claims 1 to 8 for preparing a drug for preventing or treating cancer, preferably, the cancer is any one selected from the following: urothelial carcinoma, bladder cancer, breast cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma, gastric cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, acute lymphocytic leukemia, anaplastic large cell lymphoma, multiple myeloma, prostate cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma, squamous cell carcinoma, glioblastoma, renal cell carcinoma, gastrointestinal tumors, prostate cancer, colorectal cancer, colon cancer, glioma, mesothelioma.

10. A kit of parts comprising the pharmaceutical combination according to any one of claims 1 to 8, preferably the kit is in the form of pharmaceutical dosage units.

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