Pharmaceutical combinations of anti-Nectin4 antibody drug conjugates and anti-PD-1 antibodies and uses thereof

Abstract

The present invention relates to a pharmaceutical combination of an anti-Nectin4 antibody drug conjugate and an anti-PD-1 antibody and its use for the prevention or treatment of cancer. The pharmaceutical composition disclosed by the invention is good in safety, and has a stronger anti-tumor effect compared with a single drug. The pharmaceutical composition disclosed by the invention shows a synergistic tumor inhibition effect. The invention also relates to a kit containing the pharmaceutical composition.

Description

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-PD-1 antibody or an antigen-binding fragment thereof targeting programmed death-1 (PD-1), wherein the drug combination is used for preventing or treating 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 about 66KD. 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 adhesion junctions together with cadherins.

[0003] Nectin-4 is present in high levels in normal embryonic and fetal tissues, decreases in adulthood, and has limited distribution in healthy tissues. It is highly expressed in a variety of tumor cells, such as colon cancer, ovarian cancer, cervical cancer, urothelial cancer, breast cancer, lung cancer, gastric cancer, and head and neck cancer, and mainly promotes tumor cell proliferation, differentiation, migration, and invasion by activating the PI3K / Akt pathway. Therefore, using Nectin-4 as a therapeutic target is becoming an effective strategy for treating cancers that express 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 and 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 response or clinical spillover produced by ADCs as a monotherapy in tumor treatment using antibody-drug conjugates 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 immunosuppression (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 of 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, and existing research results have shown that they can cause T cell apoptosis, immune anergy, T cell "exhaustion" and secretion of IL-10 after binding to PD-1. Studies have shown that blocking the binding of PD-1 to its ligand can restore T cell function in cancer patients. Monoclonal antibodies against PD-1 have been documented, 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 the inhibition of PD-1 on the immune surveillance of T lymphocytes with anti-tumor activity.

[0007] The biological behavior of most tumors is not dominated by a single signal transduction pathway, but multiple signal transduction pathways work together. In some cases, drugs with different mechanisms of action can be used in combination. However, any combination of drugs with different mechanisms of action but working 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 tumor occurrence mechanism, the unpredictability of the interaction between different drugs and other factors, it is still a major challenge in the medical field to find a feasible and combination drug regimen and product that can bring better effects than single drugs (reducing the single drug dose, improving the incidence and / or severity of adverse events (AE) during treatment, and / or acting in a synergistic manner, etc.). Summary of the invention

[0008] The present inventors surprisingly found that the drug combination of the present application can prevent and / or treat cancer in a synergistic manner and / or improve the incidence and / or severity of adverse events (AEs) during treatment.

[0009] 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.

[0010] Specifically, the present invention relates to the following aspects.

[0011] In a first aspect, the present invention provides a pharmaceutical combination comprising

[0012] (i) an anti-Nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof; and

[0013] (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0014] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof in the pharmaceutical combination of the present invention comprises

[0015] (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

[0016] (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;

[0017] Preferably, the anti-PD-1 antibody or antigen-binding fragment thereof comprises

[0018] (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

[0019] (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;

[0020] More preferably, the anti-PD-1 antibody comprises

[0021] (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

[0022] (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.

[0023] In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment in the drug 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.

[0024] 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, amanita chalcone, calicheamicin, eribulin and derivatives of the drugs; topoisomerase inhibitors SN38, exotecan 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.

[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, 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;

[0026] 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;

[0027] 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;

[0028] More preferably, the Ab comprises:

[0029] (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

[0030] (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;

[0031] 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.

[0032] In some embodiments, the anti-Nectin4 antibody-drug conjugate or a pharmaceutically acceptable salt thereof in the pharmaceutical combination of the present invention has a structure as shown in the following formula Ia and / or Ib:

[0033]

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

[0035] 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 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 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 glucuronidephenol.

[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 heteroaryl group, the heteroatom is N; preferably, Ar' is a substituted or unsubstituted C6 aryl group or a substituted or unsubstituted 6-membered heteroaryl group; more preferably, the antibody drug conjugate or a pharmaceutically acceptable salt thereof has the following structure:

[0039] Conjugate ADC-1:

[0040]

[0041] Conjugate ADC-2:

[0042] Conjugate ADC-3:

[0043]

[0044] Conjugate ADC-4:

[0045]

[0046] Conjugate ADC-5:

[0047]

[0048] Conjugate ADC-6:

[0049]

[0050] Conjugate ADC-7:

[0051]

[0052] In some embodiments, a pharmaceutical combination of the present invention is provided, wherein:

[0053] (i) The anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof is represented by the following formula:

[0054]

[0055] Wherein, the anti-nectin-4 antibody is as described above,

[0056] 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

[0057] (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.

[0058] In a second aspect, the present invention provides the 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.

[0059] In a third aspect, the present invention provides a kit of parts, comprising the drug combination according to the first aspect of the present invention, preferably the kit is in the form of drug dosage units.

[0060] Other aspects and embodiments of the present invention will become apparent by reference to the detailed description which follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Combined with the following Figure 1 When reading together, the preferred embodiments of the present invention described in detail below will be better understood. For the purpose of illustrating the present invention, the drawings show the currently preferred embodiments. However, it should be understood that the present invention is not limited to the precise arrangement and means of the embodiments shown in the drawings.

[0062] 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.

[0063] 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.

[0064] Figure 3 : Shows the effect of anti-Nectin4-ADC combined with PD-1 antibody Teplizumab or administered alone on tumor volume in tumor-bearing mice.

[0065] Figure 4 : Shows the effect of anti-Nectin4-ADC combined with PD-1 antibody Teplizumab or administered alone on the body weight of tumor-bearing mice. DETAILED DESCRIPTION

[0066] 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 changed. In addition, the terminology used herein is only for the purpose of describing specific embodiments and is not intended to be limiting.

[0067] I. Definitions

[0068] 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.

[0069] 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.

[0070] The term “and / or”, when used to connect two or more optional items, should be understood to mean any one of the optional items or any two or more of the optional items.

[0071] As used herein, the term "comprising" or "including" means including the stated elements, integers or steps, but does not exclude any other elements, integers or steps. In this article, when the term "comprising" or "including" is used, unless otherwise indicated, it also covers the situation consisting of the stated elements, integers or steps. For example, when referring to an antibody variable region "comprising" a specific sequence, it is also intended to cover the antibody variable region consisting of the specific sequence.

[0072] The term "antibody" is used in the broadest sense to refer to a protein that contains an antigen binding site, covering natural antibodies and artificial antibodies of various structures, including but not limited to complete antibodies and antigen-binding fragments of antibodies.

[0073] 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 subdivided into hypervariable regions (complementarity determining regions (CDRs) interspersed with more conserved regions (framework regions (FRs)). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from amino terminus to carboxyl terminus: 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 a variety of 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: Chothia numbering scheme (Chothia et al., Canonical structures for the hypervariable regions of immunoglobulins", Journal of Molecular Biology, 196, 901-917 (1987)); 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 (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 in the art or a combination thereof and human evaluation to determine the boundaries. In one embodiment, the CDR of the antibody of the present invention is a CDR sequence defined according to the Kabat numbering scheme.

[0074] Although CDRs vary from antibody to antibody, only a limited number of amino acid positions within a 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 given 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.

[0075] The term "antigen binding fragment" is a part or a section of an intact or complete antibody having fewer amino acid residues than an intact or complete antibody, which can bind to an antigen or compete with an intact antibody (i.e., an intact antibody from which the antigen binding fragment is derived) for binding to an antigen. 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, diabody, and single-domain antibody (sdAb). The Fab fragment is a monovalent fragment consisting of VL, VH, CL, and CH1 domains, for example, Fab fragments can be obtained by digesting a complete antibody with papain. In addition, digesting a complete antibody with pepsin below the disulfide bonds in the hinge region produces F(ab')2, which is a dimer of Fab' and is a divalent antibody fragment. F(ab')2 can be reduced under neutral conditions by destroying the disulfide bonds in the hinge region, thereby converting the F(ab')2 dimer into a Fab' monomer. Fab' monomers are basically Fab fragments with hinge regions (for a more detailed description of other antibody fragments, see: Fundamental Immunology, WE 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 VL and VH of the Fv fragment are encoded by independent genes, they can be connected by a synthetic linker peptide that enables the two domains to be produced as a single protein chain using a recombinant method, in which the VL region and the VH region are paired to form a single-chain Fv. The antibody fragment can be obtained by chemical methods, recombinant DNA methods, or protease digestion methods.

[0076] The term "monoclonal antibody" refers to polypeptides having substantially identical amino acid sequences or derived from the same genetic source, including antibodies and antigen-binding fragments, etc. This term also includes preparations of antibody molecules having a single molecular composition. Monoclonal antibodies exhibit a single binding specificity and affinity for a particular epitope.

[0077] 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.

[0078] The term "specific binding" and the like means that the 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.

[0079] In the context of two or more nucleic acid sequences or polypeptide sequences, the term "% identity" refers to the percent identity between sequences. If two sequences have the same amino acid sequence or nucleotide sequence in the region being compared, they are "identical". When comparing and aligning in 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 of a specified percent identity (i.e., 60% identity, optionally 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% identity in the specified region or in the entire sequence when there is no specification), then the two sequences are "substantially identical". The calculation of sequence identity between sequences is performed as follows.

[0080] 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 may be introduced in one or both of the first and second amino acid sequences or nucleic acid sequences for optimal alignment or non-homologous sequences may 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 this position.

[0081] Mathematical algorithms can be used to compare sequences and calculate percent identity between two sequences. In a preferred embodiment, the Needlema and Wunsch ((1970) J. Mol. Biol. 48: 444-453) algorithm (available at http: / / www.gcg.com) that has been integrated into the GAP program of the GCG software package is used, using a Blossum 62 matrix or a PAM250 matrix and a gap weight of 16, 14, 12, 10, 8, 6 or 4 and a length weight of 1, 2, 3, 4, 5 or 6 to determine the percent identity between two amino acid sequences. In another preferred embodiment, the GAP program in the GCG software package is used (available at http: / / www.gcg.com) to determine the percent identity between two nucleotide sequences using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70 or 80 and a length weight of 1, 2, 3, 4, 5 or 6. A particularly preferred parameter set (and the one that should be used unless otherwise stated) is the Blossum62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5.

[0082] 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, a gap penalty of 4).

[0083] Additionally or alternatively, one can further use the nucleic acid sequences and protein sequences described herein as a "query sequence" to perform a search against public databases to, for example, identify other family member sequences or related sequences.

[0084] The term "immune checkpoint" refers to a class of inhibitory signaling molecules present in the immune system, which avoid 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 escape 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, so that T lymphocytes cannot effectively play a 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).

[0085] The term "drug combination" refers to a non-fixed combination product or a fixed combination product, including but not limited to a kit, a pharmaceutical composition. The term "non-fixed combination" means that the active ingredients (e.g., (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) are administered to a patient as separate entities simultaneously, without specific time limits, or at the same or different time intervals, sequentially, wherein such administration provides preventive or therapeutically effective levels of the two active agents in the patient. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment and the anti-Nectin4 antibody drug conjugate or its pharmaceutically acceptable salt used in the drug combination are administered at levels not exceeding those 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. It is preferred that the dose and / or time interval of the two active agents be selected so that the combined use of the parts can produce an effect greater than that achieved by using any one component alone when treating a disease or condition. Each component may be in the form of a separate preparation, and the preparation forms may be the same or different.

[0086] 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 outside the gastrointestinal and local administration, usually by intravenous, and includes, but is not limited to, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, 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.

[0087] The term "dose" is the amount of a drug that elicits a therapeutic effect. Unless otherwise stated, the dose relates to the amount of the drug in free form. If the drug is in the form of a pharmaceutically acceptable salt, the amount of the drug is increased proportionally compared to the amount of the drug in free form. For example, the dose will be stated on the product packaging or in a product information leaflet.

[0088] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0089] The term "cancer" refers to a disease characterized by the proliferation of abnormal cells with 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. Examples of various cancers include, but are 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, recurrent and / or refractory cancer or cancer resistant to chemotherapy, more preferably advanced solid tumors, such as (confirmed by histology or cytology) inoperable or metastatic advanced solid tumors.

[0090] The term "inhibit" refers to a given molecule (e.g., (i) an anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof and / or (ii) an anti-PD-1 antibody or an antigen-binding fragment thereof) causing a decrease in certain parameters (e.g., Nectin-4 activity and / or PD-1 activity). For example, the term includes inhibition of at least 5%, 10%, 20%, 30%, 40% or more of the activity. Thus, inhibition need not be 100%.

[0091] The term "treatment" includes administering the drug combination of the present invention to an individual in need thereof to achieve 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 "treatment" refers to alleviating the disease (i.e., slowing down or preventing 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.

[0092] The term "preventing" includes the inhibition or delay of 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 before signs or symptoms occur in an at-risk individual.

[0093] The term "subject" refers to mammals and non-mammals. Mammals refer to any member of the mammal class, including but not limited to humans; non-human primates, cows, horses, sheep, pigs, rabbits, dogs and cats, etc. The term "subject" does not limit a particular age or gender. In some embodiments, the subject is a human.

[0094] The term "adverse event" (AE) is any unfavorable and usually 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 a treatment or expansion of immune system cells (e.g., T cells) in response to a treatment. A medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity.

[0095] 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.

[0096] 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.

[0097] All numerical ranges herein should be understood to disclose each value and numerical subset within the range, regardless of whether it is specifically disclosed in addition. For example, when any numerical range is mentioned, it should be deemed to mention each numerical value within the numerical range, such as each 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 range of numerical values.

[0098] 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.

[0099] II. Pharmaceutical Combinations of the Present Invention

[0100] The drug 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.

[0101] (i) Anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof

[0102] Antibody-drug conjugate (ADC) is 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 accurately killing tumors. Antibody-drug conjugates generally consist of three parts: antibodies or antibody-like ligands, small molecule drugs, and linkers (joints) that couple antibodies or antibody-like ligands to drugs. Due to their appropriate molecular weight, high stability, strong targeting, and low toxic side effects, antibody-drug conjugates have been considered to be the most promising anti-tumor drugs.

[0103] The anti-Nectin4 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 (joint) that couples the antibody to the small molecule drug.

[0104] In some embodiments, the anti-nectin4 antibody drug conjugate has a molecular formula of Ab-[L-CTD]m, wherein

[0105] Ab represents an anti-nectin-4 antibody or an antigen-binding fragment thereof,

[0106] L represents a connector.

[0107] CTD stands for drug,

[0108] m represents the average number of drug linkages per Ab molecule.

[0109] 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;

[0110] 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;

[0111] 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;

[0112] More preferably, the Ab comprises:

[0113] - 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

[0114] - 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;

[0115] 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.

[0116] In some embodiments, L in the molecular formula Ab-[L-CTD]m is any chemical part that can connect CTD to Ab by a covalent bond. A cross-linking agent is a bifunctional or multifunctional agent 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 agent with 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 agent. Alternatively, antibody drug conjugates 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.

[0117] In some embodiments, the CTD in the molecular formula Ab-[L-CTD]m is a cytotoxic drug. In a preferred embodiment, CTD is selected from one or more of the following: microtubule inhibitors MMAE, DM1, DM4, Tublysin, amanitin, calicheamicin, eribulin and derivatives thereof; topoisomerase inhibitors SN38, exitecan and derivatives thereof; DNA binders PBD, doxorubicin and derivatives thereof. The 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.

[0118] In some embodiments, the anti-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof has the structure shown in the following Formula Ia and / or Ib:

[0119]

[0120] Wherein: Ab is an anti-nectin-4 antibody or an antigen-binding fragment thereof as defined above;

[0121] 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 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);

[0122] 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;

[0123] 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 glucuronidephenol.

[0124] In some embodiments, L2-CTD is VcMMAE, GGFG-Dxd or VC-seco-DUBA; preferably, when Ar' is a substituted or unsubstituted 5-12-membered heteroaryl group, the heteroatom is N; preferably, Ar' is a substituted or unsubstituted C6 aryl group or a substituted or unsubstituted 6-membered heteroaryl 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.

[0125] In some embodiments, the anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof is represented by the following formula:

[0126]

[0127] Wherein, the anti-Nectin-4 antibody is as described above,

[0128] 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;

[0129] In another embodiment, the anti-Nectin4 antibody drug conjugate is PADCEV TM (Alias: enfortumabvedotin-ejfv; Chinese name: Vitin-Enfortumab), which targets the Nectin-4 protein highly expressed in urothelial carcinoma, has been approved by the FDA for the treatment of locally advanced or metastatic urothelial carcinoma. The ADC drug is composed of the human IgG1 monoclonal antibody enfortumab targeting the Nectin-4 protein and the cytotoxic agent MMAE (monomethyl auristatin E, a microtubule disruptor) (Pia M. Challita-Eid et al. (2016) Cancer Res. 76 (10): 3003-13). But PADCEV TM Can cause serious adverse skin reactions, including Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) (2019 FDA announcement).

[0130] (ii) Anti-PD-1 Antibodies or Antigen-binding Fragments thereof

[0131] The anti-PD-1 antibody or its antigen-binding fragment 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.

[0132] In one embodiment, the anti-PD-1 antibody in the pharmaceutical combination of the present invention comprises 6 CDRs, wherein HCDR1, HCDR2 and HCDR3 are composed 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 are composed 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.

[0133] 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.

[0134] 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.

[0135] Preferably, the anti-PD-1 antibody is toripalimab.

[0136] In another embodiment, the anti-PD-1 antibody in the pharmaceutical combination of the present invention comprises 6 CDRs, wherein HCDR1, HCDR2 and HCDR3 are composed 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 are composed 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 CDR sequences defined according to the Kabat numbering scheme.

[0137] 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.

[0138] 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.

[0139] Preferably, the anti-PD-1 antibody is pembrolizumab.

[0140] Other anti-PD-1 antibodies contemplated for use in the drug combinations of the present invention include MEDI0680 (U.S. Pat. No. 8,609,089), BGB-A317 (U.S. Pat. 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).

[0141] III. Uses of the pharmaceutical combination of the present invention and methods of treatment using the pharmaceutical combination of the present invention

[0142] 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.

[0143] The present invention provides a method for preventing or treating cancer, which comprises 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.

[0144] The present invention provides use of the aforementioned drug combination of the present invention in preparing a drug for preventing or treating cancer.

[0145] 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) unresectable 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).

[0146] The pharmaceutical combinations of the invention may be administered to individuals who have been treated with one or more prior therapies but have subsequently relapsed or metastasized.

[0147] The pharmaceutical combination of the present invention is administered to an individual showing elevated levels of one or more cancer-associated biomarkers [e.g., Nectin-4, programmed death ligand 1 (PD-L1)]. For example, a preventive effective amount or a therapeutic effective amount of the pharmaceutical combination of the present invention is administered to an individual with elevated levels of Nectin-4 and / or elevated levels of PD-L1.

[0148] The anti-Nectin4 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, 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.

[0149] The anti-PD-1 antibody or antigen-binding fragment thereof in the pharmaceutical combination of the present invention can be administered to an individual in need 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.

[0150] The pharmaceutical combination of the present invention can be 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, etc. Among them, the anti-Nectin4 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, and their dosage forms can be different or the same, and the anti-Nectin4 antibody drug conjugate or its pharmaceutically acceptable salt can be an intravenous dosage form or an intraperitoneal injection. The anti-PD-1 antibody or its antigen-binding fragment can be an intravenous dosage form or an intraperitoneal injection.

[0151] The pharmaceutical combination of the present invention may be a pharmaceutical dosage unit, such as a single pharmaceutical dosage unit.

[0152] 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.

[0153] 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 a patient administered a monotherapy of an anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof or a monotherapy of an anti-PD-1 antibody or an antigen-binding fragment thereof. 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 a patient administered a monotherapy of an anti-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof or a monotherapy of an anti-PD-1 antibody or an antigen-binding fragment thereof. 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-nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof, or with monotherapy with an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0154] Compared with monotherapy with anti-Nectin4 antibody drug conjugate or its pharmaceutically acceptable salt or monotherapy with anti-PD-1 antibody or its antigen-binding fragment, the drug combination of the present invention results in an increase, preferably a synergistic increase in the inhibitory effect on tumor growth. In some embodiments, compared with monotherapy with anti-Nectin4 antibody drug conjugate or its pharmaceutically acceptable salt or monotherapy with anti-PD-1 antibody or its antigen-binding fragment, 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, the 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-Nectin4 antibody drug conjugate or a pharmaceutically acceptable salt thereof or monotherapy with administration of an anti-PD-1 antibody or an antigen-binding fragment thereof, for example, 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.

[0155] 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.

[0156] 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.

[0157] IV. Kits of the Invention

[0158] 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, so that the dosage unit can be provided according to the dosage regimen or the interval of drug administration.

[0159] In one embodiment, the kit of parts of the present invention comprises in the same package:

[0160] - 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;

[0161] - A second container containing a pharmaceutical composition for administration, the pharmaceutical composition comprising an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0162] 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.

[0163] Example

[0164] 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 encompassed by the invention.

[0165] Example 1 Preparation of anti-nectin4 antibody drug conjugate

[0166] 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, and the DAR was calculated by the hydrophobic interaction chromatography (HIC) method: N = 4.0

[0167]

[0168] Example 2 In vivo efficacy of anti-Nectin4-ADC combined with anti-PD-1 antibody on tumor-bearing mice

[0169] 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 transplanted tumors in mice.

[0170] 2.1 Test drugs and materials

[0171] 2.1.1 Test drugs

[0172] (Lot number: W004692, purchased from MSD Ireland (Carlow)), which is the anti-PD-1 antibody pembrolizumab injection;

[0173] Anti-Nectin4-ADC: prepared according to Example 1.

[0174] 2.1.2 Experimental animals

[0175] Mice: SPF (Specific Pathogen Free) grade 6-8 week old female PD-1HuGEMM-C57BL / 6J mice (purchased from Shanghai Dishi Biotechnology Co., Ltd.)

[0176] 2.2 Experimental methods

[0177] Mouse colon cancer cells MC38-hNECTIN4 were used to inoculate tumors in PD-1HuGEMM-C57BL / 6J mice, 6-8 weeks old, female (purchased from Shanghai Dishi Biotechnology Co., Ltd.), thereby preparing tumor-bearing PD-1HuGEMM-C57BL / 6J mice.

[0178] 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 into the right hind subcutaneous part of PD-1HuGEMM-C57BL / 6J mice with a 1mL syringe, 100 μL / mouse, and each mouse was inoculated with approximately 5.0×10 6 The MC38-hNECTIN4 tumor cells were used to establish the PD-1 tumor-bearing HuGEMM-C57BL / 6J mouse model.

[0179] This experiment used anti-Nectin4-ADC (prepared with reference to Example 1) and anti-PD-1 antibody (Batch number: W004692, purchased from MSD Ireland (Carlow)) for group comparison study (as shown in Table 1). The day of grouping was day 0, and the mice were given the first injection of drug on the day of grouping, with a volume of 10 μL / g; the solvent group was given the same volume of solvent (normal saline); the specific dosage and dosing schedule are shown in Table 1. The tumor volume was measured twice a week, the mice were weighed, and the data were recorded. The experiment was terminated, the experimental endpoint was reached, or the tumor volume reached 3000mm 3The animals were killed by CO2 anesthesia, and the tumors were subsequently dissected and photographed.

[0180] Table 1. Dosage and dosing schedule

[0181]

[0182] 2.3 Experimental Results and Conclusions

[0183] Results Figure 1 and Table 2. There was no significant difference in each single drug group compared with the saline control group. However, compared with the saline control group, the two combined drug groups, anti-Nectin4-ADC 2mg / kg+Keytruda 3mg / kg group and anti-Nectin4-ADC 5mg / kg+Keytruda 3mg / kg group, showed significant differences (p1 value <0.05 and p1 value <0.001, respectively).

[0184] The anti-tumor effect of the anti-Nectin4-ADC 2mg / kg + Keytruda 3mg / kg combined administration 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).

[0185] The anti-tumor effect of the anti-Nectin4-ADC 5mg / kg+Keytruda 3mg / kg combined administration group was better than that of the anti-Nectin4-ADC 10mg / kg single-drug group and the Keytruda 3mg / kg single-drug group, and there were significant differences compared with each single-drug group (p3 values ​​<0.01). Although the total dose of the combined drug group (anti-Nectin4-ADC 5mg / kg+Keytruda 3mg / kg) was less than that of the single-drug group (anti-Nectin4-ADC 10mg / kg), it showed a significantly better efficacy than the single-drug group. It can be seen that the combination of anti-Nectin4-ADC and Keytruda showed a significant synergistic therapeutic effect.

[0186] In this experiment, there was no significant weight loss in the mice in the saline control group, anti-Nectin4-ADC 5, 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 ( Figure 2 ). One mouse in group 2 and one mouse in group 4 died on the 19th day and the 15th day due to tumor rupture, respectively.

[0187] Table 2. Efficacy analysis based on tumor volume of each group of mice in the subcutaneous mouse colon cancer model MC38-hNectin4

[0188]

[0189] Note: a. The data of tumor volume are expressed as “mean ± standard error” (Mean ± SEM);

[0190] bT / C%=T / C×100%; TGI%=(1-T / C)×100%, where T and C are the absolute tumor volumes (Vt-V0) of each model treatment group and the saline control group on the 19th day of the experiment.

[0191] Example 3 In vivo efficacy of anti-Nectin4-ADC combined with anti-PD-1 antibody on tumor-bearing mice (II)

[0192] 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 transplanted tumors in mice.

[0193] 3.1 Test drugs and materials

[0194] 3.1.1 Test drugs

[0195] (Purchased from Junshi Biosciences), the main ingredient of this product is Toripalimab, an anti-PD-1 antibody.

[0196] Anti-Nectin4-ADC: prepared according to Example 1.

[0197] (Enfortumab Vedotin): White lyophilized powder, 30 mg / bottle, batch number 102374, stored at 2-8°C away from light. It is an ADC drug that is a nectin-4 targeting antibody and a microtubule inhibitor.

[0198] 3.1.2 Experimental animals

[0199] Mice: SPF 6-week-old female PD-1-HU mice (purchased from Shanghai Nanmo Biotech Co., Ltd.).

[0200] 3.2 Experimental methods

[0201] Tumor-bearing PD-1-HU mice were prepared by inoculating tumors with mouse colon cancer cells MC38 / Nectin4 hPD-L1, 6 weeks old, female (purchased from Shanghai Nanmo Biotechnology Co., Ltd.).

[0202] Specifically, a tumor-bearing mouse model was established using mouse colon cancer cells MC38 / Nectin4 hPD-L1 (Shanghai South Model Organisms Technology Co., Ltd.). MC38 / Nectin4hPD-L1 cells were cultured with 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 subcutaneously in the right hindquarters of PD-1-HU mice with a 1mL syringe, 100 μL / mouse, and each mouse was inoculated with approximately 5.0×10 5 MC38 / Nectin4hPD-L1 tumor cells were used to establish a tumor-bearing PD-1-HU mouse model.

[0203] In this experiment, anti-Nectin4-ADC and anti-PD-1 antibody TuoYi (purchased from Junshi Biosciences) were used for group comparison (as shown in Table 3). 3 Afterwards, the animals were randomly divided into groups according to the tumor volume, and the day of grouping was 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. The tumor volume was measured twice a week, the mice were weighed, and the data were recorded. The experiment was completed, the experimental endpoint was reached, or the tumor volume reached 3000mm 3 The animals were killed by CO2 anesthesia, and the tumors were subsequently dissected and photographed.

[0204] Table 3. Dosage and dosing schedule

[0205]

[0206] Note: IP- intraperitoneal injection, administration started at D0; BIW: twice a week.

[0207] Results Figure 3 , Figure 4And Table 4. Compared with the solvent group, the anti-Nectin4-ADC 3mg / kg monotherapy group, the Tuoyi 5mg / kg monotherapy group, the two combined administration groups of anti-Nectin4-ADC 3mg / kg + Tuoyi 5mg / kg group and PADCEV 3mg / kg + Tuoyi 5mg / kg group all had certain therapeutic effects on MC38 / Nectin4 hPD-L1 subcutaneous transplanted tumors. Compared with the Tuoyi monotherapy group, the two combined administration groups of anti-Nectin4-ADC 3mg / kg + Tuoyi 5mg / kg group and PADCEV 3mg / kg + Tuoyi 5mg / kg group had better anti-tumor effects. The anti-tumor effect of the combined administration group of anti-Nectin4-ADC 3mg / kg + Tuoyi 5mg / kg group was better than that of PADCEV 3mg / kg + Tuoyi 5mg / kg group. The anti-tumor effect of the anti-Nectin4-ADC 3mg / kg + Tuoyi 5mg / kg group was better than that of the anti-Nectin4-ADC 3mg / kg monotherapy group. The tumor-bearing mice tolerated the above drugs well, without obvious symptoms such as weight loss, and no animals died during the experiment.

[0208] Table 4. Efficacy on mouse colon cancer MC38 / Nectin4 hPD-L1 subcutaneous transplanted tumors

[0209]

[0210] Note: TV: tumor volume, mm 3 .

[0211] Example 4 Treatment of urothelial carcinoma with anti-Nectin4-ADC combined with anti-PD-1 antibody

[0212] This study is a phase Ib / II open-label, single-arm, multicenter study in patients with locally advanced or metastatic urothelial carcinoma. The study includes a screening phase for patient enrollment, a treatment phase of up to 2 years, and a safety / efficacy follow-up phase.

[0213] 1. Clinical research objectives:

[0214] The primary objective is to evaluate the safety and tolerability of anti-nectin4-ADC combined with toripalimab in patients with locally advanced or metastatic urothelial carcinoma. Secondary objectives include the following:

[0215] 1) To evaluate the preliminary efficacy of anti-Nectin4-ADC combined with toripalimab in patients with locally advanced or metastatic urothelial carcinoma;

[0216] 2) Evaluate the pharmacokinetic (PK) characteristics of anti-Nectin4-ADC;

[0217] 3) evaluate the immunogenicity of anti-Nectin4-ADC;

[0218] 4) Evaluate the relationship between biomarkers Nectin-4 and PD-L1 in tumor tissue and efficacy or prognosis.

[0219] The study protocol and all amendments were approved by the institutional ethics committees of all participating centers.

[0220] 2. Test drug:

[0221] The active components of the test drug described in this example are: anti-Nectin4-ADC and anti-PD-1 antibody.

[0222] Anti-Nectin4-ADC: prepared with reference to Example 1, specification is 20 mg / bottle, and is a conjugate of anti-Nectin-4 antibody and MMAE;

[0223] The anti-PD-1 antibody is Teplizumab injection, produced by Shanghai Junshi Biotechnology Co., Ltd.

[0224] 3. Patient inclusion criteria

[0225] 1) All subjects or guardians must personally sign a written informed consent form approved by the ethics committee before starting any screening procedure;

[0226] 2) Age between 18 and 80 years old (including the cutoff value) when signing the informed consent form, regardless of gender;

[0227] 3) Eastern Cooperative Oncology Group (ECOG) score of 0 to 1;

[0228] 4) Locally advanced or metastatic urothelial carcinoma (including bladder, ureter, renal pelvis and urethra) confirmed by histopathology, with squamous differentiation, glandular differentiation or mixed type of urothelial carcinoma also meets the requirements, and radical surgical resection is not possible;

[0229] 5) Previous treatment for locally advanced or metastatic urothelial carcinoma: previous standard treatment failure or no previous systemic treatment;

[0230] 6) Must provide archived tumor tissue samples or undergo new tumor tissue biopsy;

[0231] 7) The expected survival period is not less than 12 weeks;

[0232] 8) According to the RECIST v1.1 standard, there is at least one measurable lesion during the screening period; for patients with a clear history of radiotherapy, the measurable lesion has shown clear progression outside the radiation field or after completion of radiotherapy;

[0233] 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 blood test must be performed ≥7 days after any growth factor therapy or ≥14 days after any blood transfusion:

[0234] Absolute neutrophil count (ANC) ≥ 1.5 × 10 9 / L;

[0235] Platelet count (PLT) ≥100×10 9 / L;

[0236] Hemoglobin (Hb) ≥ 90 g / L;

[0237] Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ≤ 3 times the upper limit of normal (ULN);

[0238] Total bilirubin ≤ 1.5 times ULN (total bilirubin should be < 3 times ULN for subjects with Gilbert's syndrome);

[0239] International Normalized Ratio (INR) ≤ 1.5 times ULN;

[0240] Creatinine clearance ≥50 mL / min (calculated according to the Cockcroft-Gault formula), or serum creatinine ≤1.5 times ULN.

[0241] 10) Male or female subjects must meet any of the following conditions:

[0242] Women of infertile potential, especially those who have undergone hysterectomy, bilateral oophorectomy, bilateral tubal ligation or are postmenopausal;

[0243] 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 within 180 days after the end of medication, such as double barrier contraceptive methods, condoms, oral or injectable contraceptives, intrauterine devices, etc.;

[0244] Male subjects who have undergone vasectomy or agree to use contraceptive measures during the study treatment phase and within 180 days of the last dose of study drug.

[0245] 11) Able to understand and perform planned visits, treatments, laboratory tests, and other study procedures.

[0246] Exclusion criteria:

[0247] 1) Received chemotherapy, radiotherapy or other anti-tumor treatment within 21 days before the first study drug treatment;

[0248] 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 that the investigator judged would not affect participation in the trial (such as tooth extraction, puncture, etc.);

[0249] 3) (Cohort expansion phase only) Patients who have previously used PD-1, PD-L1, or PD-L2 inhibitors for locally advanced or metastatic urothelial carcinoma, including but not limited to pembrolizumab, atezolizumab, and tislelizumab;

[0250] 4) Patients who have received MMAE-conjugated antibody-drug therapy before, such as Enfortumab vedotin;

[0251] 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);

[0252] 6) Subjects have grade 2 or higher peripheral neuropathy;

[0253] 7) Subjects with poor blood sugar control;

[0254] 8) Investigators assess subjects who may have an increased risk of corneal disease before the first dose;

[0255] 9) Subjects who had clinically significant cardiovascular and cerebrovascular diseases within 6 months before the first study medication, including but not limited to: ① acute myocardial infarction; ② unstable angina pectoris; ③ cerebrovascular accident (except 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, torsade de pointes); ⑤ New York Heart Association (NYHA) 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 found by echocardiography (ECHO); ⑧ hypertensive subjects who are not well controlled by drug treatment; ⑨ other arrhythmias that are judged by the researchers to be unsuitable for inclusion in this study;

[0256] 10) Active infection, including but not limited to: hepatitis B virus infection (HBsAg must be positive and HBV-DNA>500IU / mL), hepatitis C virus infection (HCV-Ab must be positive and HCV-RNA must be positive), human immunodeficiency virus infection (HIV-Ab positive), active tuberculosis infection, or other active infections requiring systemic treatment within 14 days before the first study medication (except for routine preventive anti-infection treatment);

[0257] 11) Subjects have other severe or uncontrolled diseases that the investigators believe are not suitable for participation in this clinical trial or may affect the subjects' 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;

[0258] 12) Subjects have 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, imaging examinations have confirmed no disease progression within 4 weeks before the first dose of the study, and 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 at least 28 days before the first dose of study treatment. Carcinomatous meningitis should be excluded regardless of whether their clinical condition is stable;

[0259] 13) The subject has previously received allogeneic hematopoietic stem cell transplantation or solid organ transplantation;

[0260] 14) The subject has a history of drug abuse or suffers from mental illness, which affects the trial compliance;

[0261] 15) Those who are known to be allergic or intolerant to the study drug or any of its components;

[0262] 16) Subjects who have used P-glycoprotein (P-gp) inhibitors or inducers, strong CYP3A4 inhibitors or inducers within 14 days before the first study medication;

[0263] 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;

[0264] 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;

[0265] 19) The subject has received / used other clinical research drugs or experimental medical devices within 28 days before the first study medication;

[0266] 20) Active autoimmune diseases requiring systemic treatment within 2 years before the first study medication;

[0267] 21) Subjects who have other malignant tumors within 3 years before the first study drug use, except for 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 removed, such as cervical carcinoma in situ, breast ductal carcinoma in situ, etc.;

[0268] 22) Other situations that the researcher judges are not suitable for this study.

[0269] Patients who met the above criteria were enrolled and received anti-Nectin4-ADC combined with toripalimab treatment.

[0270] 4. Clinical dosing regimen

[0271] The enrolled subjects received anti-Nectin4-ADC by intravenous infusion on day 1 (D1) and day 8 (D8) of each cycle, and Teplizumab was administered by intravenous infusion on D1 of each cycle, with each cycle consisting of 21 days. The doses of anti-Nectin4-ADC in the dose exploration phase were 1.0 mg / kg and 1.25 mg / kg, respectively, and in the cohort expansion phase, the dose was 1.25 mg / kg. The dose of Teplizumab was 240 mg. All subjects received anti-Nectin4-ADC combined with Teplizumab 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.

[0272] 5. Results and Summary

[0273] Antitumor activity

[0274] For the combination therapy, the following clinical results have been obtained. A total of 15 subjects completed at least one post-baseline tumor assessment. All subjects' target lesions shrank to varying degrees, and all patients with PR efficacy were treated in the same group. Among them, 4 evaluable patients who had previously received platinum-containing chemotherapy and PD-1 / L1 treatment and received anti-Nectin4-ADC (1.25 mg / kg) combination therapy in this study had an ORR of 75% (3 / 4).

[0275] 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 treatment, 37 patients were evaluable and the ORR was 62% (23 / 37).

[0276] 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).

[0277] Safety Assessment

[0278] In the anti-Nectin4-ADC (1.25 mg / kg) combination treatment group, common TRAEs (all grades with an incidence ≥ 10%) were rash, anemia, increased aspartate aminotransferase, increased serum 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.

[0279] Only one subject among all the subjects had to reduce the dose of anti-Nectin4-ADC due to adverse events. No TRAEs leading to death occurred. These results show that the combination of anti-Nectin4-ADC and anti-PD-1 antibody has good anti-urothelial cancer efficacy.

[0280] 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 may 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.

[0281] Exemplary sequences

[0282]

[0283]

[0284]

[0285]

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: and / or 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 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); 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: and / or 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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