Combination therapy of an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent for treating cancer

Abstract

The present disclosure provides a combination therapy comprising (i) an active agent that binds to PD-1 (e.g., an anti-PD-1 antibody), (ii) an active agent that binds to TIM-3 (e.g., an anti-TIM-3 antibody), and / or (iii) an active agent that binds to LAG-3 (e.g., an anti-LAG-3 antibody). The present disclosure provides its pharmaceutical composition, its use, and a treatment method comprising administration of the combination therapy to a subject, including a method of treating cancer.

Description

Technical Field

[0001] (Related Applications) This application claims priority to U.S. Provisional Patent Application No. 63 / 349,876, filed on June 7, 2022, and U.S. Provisional Patent Application No. 63 / 405,243, filed on September 9, 2022. The entire contents of these applications are incorporated herein by reference.

[0002] (Sequence Listing) This application includes a sequence listing that was electronically submitted in XML format, the entire contents of which are incorporated herein by reference. The XML copy created on June 6, 2023, is named 12092-0051-00304_SL.XML and is 57,780 bytes in size.

[0003] (Field of Disclosure) The present disclosure provides combination therapies comprising (i) an active agent that binds to PD-1 (e.g., an anti-PD-1 antibody), (ii) an active agent that binds to TIM-3 (e.g., an anti-TIM-3 antibody), and / or (iii) an active agent that binds to LAG-3 (e.g., an anti-LAG-3 antibody). The present disclosure provides pharmaceutical compositions, uses thereof, kits thereof, and methods of treatment comprising administration of the combination therapy to a subject, including methods of treating cancer.

[0004] (Disclosure) Considerable focus has been placed on understanding the activity of the immune system within the tumor microenvironment and identifying the mechanisms of tumor resistance to immune surveillance. Loss of CD8+ T cell effector function is one such proposed mechanism. This loss of function, or T cell exhaustion, has been reported to be due in part to the accumulation and increased diversity of inhibitory receptors, or checkpoints, on the cell surface of CD8+ T cells (Wherry et al., Nat Immunol (2011); 12:492-499).

[0005] Antibodies directed against the PD-1 / PDL-1 axis have improved cancer treatment, but the majority of clinical patients still do not respond to these agents or have lost their response. Patients who relapse or are resistant to therapy with PD-1 inhibitors represent a high unmet need in oncology. Early clinical evidence of the effectiveness of combination therapies in patients who have failed anti-PDL-1 therapy is accumulating. Combining blockade of the PDL-1 axis with blockade of either LAG-3 or TIM-3 has resulted in only modest responses in patients who did not respond to anti-PD-1 therapy alone (Ascierto et al., Ann Oncol (2017; 28(suppl 5):v605-v649; Davar et al., Society for Immunotherapy of Cancer (2018); November 7-11, 2018; Washington, DC. Abstract O21).

[0006] Therefore, it remains difficult to intervene in and disrupt the process of T cell exhaustion by using inhibition by the multiplexing of checkpoints along the exhaustion hierarchy. Potentially, multiple checkpoint inhibitors are required to restore anti-tumor T cell effector function in situ. Thus, there is a need for improved combination therapies for the treatment of PD-1-related diseases such as cancer. SUMMARY OF THE INVENTION

[0007] Details of various embodiments of the present disclosure are set forth in the following description. In certain embodiments, the present disclosure provides a combination therapy comprising at least two (e.g., three) of a therapeutically effective amount of an anti-PD-1 active agent, a therapeutically effective amount of an anti-TIM-3 active agent, and a therapeutically effective amount of an anti-LAG-3 active agent. In certain embodiments, the combination therapy comprises at least two (e.g., three) of: (i) an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; (ii) an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; and (iii) an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the combination therapy comprises (a) an anti-PD-1 active agent and an anti-TIM-3 active agent, (b) an anti-PD-1 active agent and an anti-LAG-3 active agent, (c) an anti-TIM-3 active agent and an anti-LAG-3 active agent, or (d) an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent. In certain embodiments, the combination therapy comprises an anti-PD-1 active agent and an anti-TIM-3 active agent. In certain embodiments, the combination therapy comprises an anti-PD-1 active agent and an anti-LAG-3 active agent. In certain embodiments, the combination therapy comprises an anti-TIM-3 active agent and an anti-LAG-3 active agent. In certain embodiments, the combination therapy comprises an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent.

[0008] In certain embodiments, the combination therapy is for use in treating cancer in a human subject in need thereof. In certain embodiments, the combination therapy is for use in a method of treating cancer in a human subject in need thereof. In certain embodiments, the combination therapy is for use in the manufacture / production of a medicament for use in treating cancer in a human subject in need thereof.

[0009] In certain embodiments, the present disclosure provides an anti-PD-1 active agent for use in the treatment of cancer in a human subject in need thereof, the anti-PD-1 active agent being administered in a therapeutically effective amount, simultaneously or sequentially with at least one (e.g., both) of a therapeutically effective amount of an anti-TIM-3 active agent and a therapeutically effective amount of an anti-LAG-3 agent. In certain embodiments, the anti-PD-1 active agent is administered at a dose of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, (i) with an anti-TIM-3 active agent at a dose of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, and (ii) with at least one (e.g., both) of an anti-LAG-3 active agent at a dose of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, simultaneously or sequentially. In certain embodiments, the anti-PD-1 active agent is administered at a dose of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, once every three weeks or once every four weeks, (i) with an anti-TIM-3 active agent administered once every two weeks, once every three weeks or once every four weeks at a dose of about 300 mg to about 1000 mg, in some embodiments about 400 to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, and (ii) with at least one (e.g., both) of an anti-LAG-3 active agent administered once every two weeks, once every three weeks or once every four weeks at a dose of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg.In certain embodiments, the anti-PD-1 agent is administered simultaneously or sequentially with (a) an anti-TIM-3 agent, (b) an anti-LAG-3 agent, or (c) both an anti-TIM-3 agent and an anti-LAG-3 agent. In certain embodiments, the anti-PD-1 agent is administered simultaneously or continuously with the anti-TIM-3 agent. In certain embodiments, the anti-PD-1 agent is administered simultaneously or sequentially with the anti-LAG-3 agent. In certain embodiments, the anti-PD-1 agent is administered simultaneously or sequentially with both an anti-TIM-3 agent and an anti-LAG-3 agent.

[0010] In certain embodiments, the present disclosure provides an anti-PD-1 agonist, an anti-TIM-3 agonist, and an anti-LAG-3 agent for use in the treatment of cancer in a human subject in need thereof, wherein the anti-PD-1 agonist is administered in a therapeutically effective amount, the anti-TIM-3 agonist is administered in a therapeutically effective amount, and the anti-LAG-3 agonist is administered in a therapeutically effective amount. In certain embodiments, (i) the anti-PD-1 agonist is administered at a dose of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; (ii) the anti-TIM-3 agonist is administered at a dose of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; (iii) the anti-LAG-3 agonist is administered at a dose of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg.In certain embodiments, (i) the anti-PD-1 agent is administered at a dose of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, once every 3 to 5 weeks, and in some embodiments once every 3 weeks or once every 4 weeks; (ii) the anti-TIM-3 agent is administered at a dose of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks; (iii) the anti-LAG-3 agent is administered at a dose of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks. In certain embodiments, the present disclosure provides for the use of an anti-PD-1 agent, an anti-TIM-3 agent, and an anti-LAG-3 agent in the treatment of cancer in a human subject in need thereof. In certain embodiments, the present disclosure provides for the use of an anti-PD-1 agent, an anti-TIM-3 agent, and an anti-LAG-3 agent in the manufacture / production of a medicament for use in the treatment of cancer in a human subject in need thereof.

[0011] In certain embodiments, the present disclosure provides a method of treating cancer in a human subject in need thereof, the method comprising administering to the subject at least two (e.g., three) of (i) a therapeutically effective amount of an anti-PD-1 agent, (ii) a therapeutically effective amount of an anti-TIM-3 agent, and (iii) a therapeutically effective amount of an anti-LAG-3 agent. In certain embodiments, the method comprises administering to the subject a combination therapy agent comprising at least two (e.g., three) of (i) from about 300 mg to about 600 mg, in some embodiments from about 375 mg to about 550 mg, in some embodiments from about 375 mg to about 500 mg, in some embodiments from about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, of an anti-PD-1 agent; (ii) from about 300 to about 1000 mg, in some embodiments from about 400 to about 1000 mg, in some embodiments from about 350 to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, of an anti-TIM-3 agent; and (iii) from about 300 mg to about 800 mg, in some embodiments from about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, of an anti-LAG-3 agent.In certain embodiments, the method comprises administering to a subject at least two (e.g., three) of: (i) an anti-PD-1 active agent at a dose of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg once every 3 to 5 weeks; and in some embodiments once every 3 weeks or once every 4 weeks; (ii) an anti-TIM-3 active agent at a dose of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg, or about 1000 mg once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks; and (iii) an anti-LAG-3 active agent at a dose of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg, or about 750 mg once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks. In certain embodiments, the method comprises: (a) administering an anti-PD-1 active agent and an anti-TIM-3 active agent to the subject; (b) administering an anti-PD-1 active agent and an anti-LAG-3 active agent to the subject; (c) administering an anti-TIM-3 active agent and an anti-LAG-3 active agent to the subject; or (d) administering an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent to the subject. In certain embodiments, the method comprises administering an anti-PD-1 active agent and an anti-TIM-3 active agent to the subject. In certain embodiments, the method comprises administering an anti-PD-1 active agent and an anti-LAG-3 active agent to the subject. In certain embodiments, the method comprises administering an anti-TIM-3 active agent and an anti-LAG-3 active agent to the subject. In certain embodiments, the method comprises administering an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent to the subject.

[0012] In certain embodiments, the present disclosure provides a kit for treating cancer in a human subject in need thereof, the kit comprising at least two (e.g., three): (i) an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; (ii) an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; and (iii) an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, for co-administration. In certain embodiments, the kit further comprises instructions for administering the anti-PD-1 active agent (e.g., once every 3 weeks or once every 4 weeks), instructions for administering the anti-TIM-3 active agent (e.g., once every 2 weeks, once every 3 weeks or once every 4 weeks), and / or instructions for administering the anti-LAG-3 active agent (e.g., once every 2 weeks, once every 3 weeks or once every 4 weeks).

[0013] In certain embodiments, the methods, co-therapies, uses, or kits of the present disclosure comprise an anti-PD-1 active agent. In certain embodiments, the anti-PD-1 active agent comprises an anti-PD-1 antibody or a PD-1 binding fragment thereof. In certain embodiments, the anti-PD-1 antibody is retifanlimab (also known as MGA012 and INCMGA00012, CAS Reg No. 2079108-44-2). In certain embodiments, the anti-PD-1 active agent is administered intravenously. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every 3 weeks or once every 4 weeks. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every 3 weeks. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every 4 weeks.

[0014] In certain embodiments, the anti-PD-1 agent comprises an anti-PD-1 antibody or a PD-1 binding fragment thereof, which comprises (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain. In certain embodiments, the CDRH1 domain has the amino acid sequence SYWMN (SEQ ID NO: 1). In certain embodiments, the CDRH2 domain has the amino acid sequence VIHPSDSETWLDQKFK (SEQ ID NO: 2). In certain embodiments, the CDRH3 domain has the amino acid sequence EHYGTSPFAY (SEQ ID NO: 3). In certain embodiments, the CDRH1 domain has the amino acid sequence SYWMN (SEQ ID NO: 1), the CDRH2 domain has the amino acid sequence VIHPSDSETWLDQKFK (SEQ ID NO: 2), and the CDRH3 domain has the amino acid sequence EHYGTSPFAY (SEQ ID NO: 3). In certain embodiments, the CDRL1 domain has the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 4). In certain embodiments, the CDRL2 domain has the amino acid sequence AASNQGS (SEQ ID NO: 5). In certain embodiments, the CDRL3 domain has the amino acid sequence QQSKEVPYT (SEQ ID NO: 6). In certain embodiments, the CDRL1 domain has the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 4), the CDRL2 domain has the amino acid sequence AASNQGS (SEQ ID NO: 5), and the CDRL3 domain has the amino acid sequence QQSKEVPYT (SEQ ID NO: 6). In certain embodiments, the CDRH1 domain has the amino acid sequence SYWMN (SEQ ID NO: 1), the CDRH2 domain has the amino acid sequence VIHPSDSETWLDQKFK (SEQ ID NO: 2), the CDRH3 domain has the amino acid sequence EHYGTSPFAY (SEQ ID NO: 3), the CDRL1 domain has the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 4), the CDRL2 domain has the amino acid sequence AASNQGS (SEQ ID NO: 5), and the CDRL3 domain has the amino acid sequence QQSKEVPYT (SEQ ID NO: 6).

[0015] In certain embodiments, the heavy chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 7). In certain embodiments, the light chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIK (SEQ ID NO: 8). In certain embodiments, the heavy chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 7), and the light chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIK (SEQ ID NO: 8).

[0016] In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 9). In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 9), and the anti-PD-1 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0017] In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, or IgG4 isotype. In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises an Fc region of the IgG4 isotype. In certain embodiments, the antibody comprises a hinge domain of the IgG4 isotype that contains a stabilizing mutation. In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises (A) one or more amino acid modifications that decrease the affinity of the variant Fc region for FcγR, wherein the one or more modifications that decrease the affinity of the variant Fc region for FcγR include the substitutions L234A, L235A, or L234A+L235A, where the numbering is that of the EU index as in Kabat, and / or (B) one or more amino acid modifications that enhance the serum half-life of the variant Fc region, wherein the one or more modifications that enhance the serum half-life of the variant Fc region include the substitutions M252Y, M252Y+S254T, M252Y+T256E, M252Y+S254T+T256E, or K288D+H435K, where the numbering is that of the EU index in Kabat.

[0018] In certain embodiments, the method, combination therapy, use, or kit of the present disclosure comprises an anti-TIM-3 active agent. In certain embodiments, the anti-TIM-3 active agent comprises an anti-TIM-3 antibody or a TIM-3 binding fragment thereof. In certain embodiments, the anti-TIM-3 antibody is Antibody A described herein. In certain embodiments, the anti-TIM-3 active agent is administered intravenously. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every two weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every three weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every four weeks.

[0019] In certain embodiments, the anti-TIM-3 active agent comprises an anti-TIM-3 antibody or a TIM-3 binding fragment thereof comprising (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain. In certain embodiments, the CDRH1 domain has the amino acid sequence RQNAWS (SEQ ID NO: 11). In certain embodiments, the CDRH2 domain has the amino acid sequence WVSAISGSGGSTY (SEQ ID NO: 12). In certain embodiments, the CDRH3 domain has the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 13). In certain embodiments, the CDRH1 domain has the amino acid sequence RQNAWS (SEQ ID NO: 11), the CDRH2 domain has the amino acid sequence WVSAISGSGGSTY (SEQ ID NO: 12), and the CDRH3 domain has the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 13). In certain embodiments, the CDRL1 domain has the amino acid sequence RASQSVSSYLA (SEQ ID NO: 14). In certain embodiments, the CDRL2 domain has the amino acid sequence DASNRAT (SEQ ID NO: 15). In certain embodiments, the CDRL3 domain has the amino acid sequence QQYGSSPLT (SEQ ID NO: 16). In certain embodiments, the CDRL1 domain has the amino acid sequence RASQSVSSYLA (SEQ ID NO: 14), the CDRL2 domain has the amino acid sequence DASNRAT (SEQ ID NO: 15), and the CDRL3 domain has the amino acid sequence QQYGSSPLT (SEQ ID NO: 16). In certain embodiments, the CDRH1 domain has the amino acid sequence RQNAWS (SEQ ID NO: 11), the CDRH2 domain has the amino acid sequence WVSAISGSGGSTY (SEQ ID NO: 12), the CDRH3 domain has the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 13), the CDRL1 domain has the amino acid sequence RASQSVSSYLA (SEQ ID NO: 14), the CDRL2 domain has the amino acid sequence DASNRAT (SEQ ID NO: 15), and the CDRL3 domain has the amino acid sequence QQYGSSPLT (SEQ ID NO: 16).

[0020] In certain embodiments, the heavy chain variable domain of the anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSS (SEQ ID NO: 17). In certain embodiments, the light chain variable domain of the anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK (SEQ ID NO: 18). In certain embodiments, the heavy chain variable domain of the anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSS (SEQ ID NO: 17), and the light chain variable domain of the anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK (SEQ ID NO: 18).

[0021] In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 19). In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 19), and the anti-TIM-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).

[0022] In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of an IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of human IgG1 isotype, and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of human IgG4 isotype, and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-TIM-3 antibody or fragment thereof is antagonist to human TIM-3. In certain embodiments, the anti-TIM-3 antibody or fragment thereof inactivates, reduces, or inhibits the activity of human TIM-3. In certain embodiments, the anti-TIM-3 antibody or fragment thereof inhibits the binding of human TIM-3 to phosphatidylserine.

[0023] In certain embodiments, the method, combination therapy, use, or kit of the present disclosure comprises an anti-LAG-3 active agent. In certain embodiments, the anti-LAG-3 active agent comprises an anti-LAG-3 antibody or a LAG-3 binding fragment thereof. In certain embodiments, the anti-LAG-3 antibody is Antibody B (described herein). In certain embodiments, the anti-LAG-3 active agent is administered intravenously. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every two weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every three weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every four weeks.

[0024] In certain embodiments, the anti-LAG-3 agent comprises an anti-LAG-3 antibody or a LAG-3 binding fragment thereof, which comprises (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain. In certain embodiments, the CDRH1 domain has the amino acid sequence DTYIH (SEQ ID NO: 21). In certain embodiments, the CDRH2 domain has the amino acid sequence EIDPANDNTKYDPKFQG (SEQ ID NO: 22). In certain embodiments, the CDRH3 domain has the amino acid sequence YYYKYDVGGFDY (SEQ ID NO: 23). In certain embodiments, the CDRH1 domain has the amino acid sequence DTYIH (SEQ ID NO: 21), the CDRH2 domain has the amino acid sequence EIDPANDNTKYDPKFQG (SEQ ID NO: 22), and the CDRH3 domain has the amino acid sequence YYYKYDVGGFDY (SEQ ID NO: 23). In certain embodiments, the CDRL1 domain has the amino acid sequence SVSSSISSSNLH (SEQ ID NO: 24). In certain embodiments, the CDRL2 domain has the amino acid sequence GTSNLAS (SEQ ID NO: 25). In certain embodiments, the CDRL3 domain has the amino acid sequence QQWSSYPFT (SEQ ID NO: 26). In certain embodiments, the CDRL1 domain has the amino acid sequence SVSSSISSSNLH (SEQ ID NO: 24), the CDRL2 domain has the amino acid sequence GTSNLAS (SEQ ID NO: 25), and the CDRL3 domain has the amino acid sequence QQWSSYPFT (SEQ ID NO: 26). In certain embodiments, the CDRH1 domain has the amino acid sequence DTYIH (SEQ ID NO: 21), the CDRH2 domain has the amino acid sequence EIDPANDNTKYDPKFQG (SEQ ID NO: 22), the CDRH3 domain has the amino acid sequence YYYKYDVGGFDY (SEQ ID NO: 23), the CDRL1 domain has the amino acid sequence SVSSSISSSNLH (SEQ ID NO: 24), the CDRL2 domain has the amino acid sequence GTSNLAS (SEQ ID NO: 25), and the CDRL3 domain has the amino acid sequence QQWSSYPFT (SEQ ID NO: 26).

[0025] In certain embodiments, the heavy chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSS (SEQ ID NO: 27). In certain embodiments, the light chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIK (SEQ ID NO: 28). In certain embodiments, the heavy chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSS (SEQ ID NO: 27), and the light chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIK (SEQ ID NO: 28).

[0026] In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 29). In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 29), and the anti-LAG-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).

[0027] In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the human IgG1 isotype, and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the human IgG4 isotype, and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-LAG-3 antibody or fragment thereof is antagonist to human LAG-3. In certain embodiments, the anti-LAG-3 antibody or fragment thereof inactivates, reduces, or inhibits the activity of human LAG-3. In certain embodiments, the anti-LAG-3 antibody or fragment thereof inhibits the binding of human LAG-3 to MHC class II.

[0028] In certain embodiments, the anti-PD-1 agent is administered prior to the anti-TIM-3 agent and the anti-LAG-3 agent. In certain embodiments, the anti-TIM-3 agent is administered prior to the anti-LAG-3 agent. In certain embodiments, the anti-LAG-3 agent is administered prior to the anti-TIM-3 agent. In certain embodiments, the anti-TIM-3 agent and the anti-LAG-3 agent are administered simultaneously.

[0029] In certain embodiments, the anti-PD-1 active agent is in an acetate, sucrose, polysorbate 80 (“PS80”), water, and has a pH of from about 4.0 to about 6.5, and optionally, the concentration of the anti-PD-1 agent in the pharmaceutical composition is from about 10 mg / mL to about 100 mg / mL. In certain embodiments, the anti-TIM-3 active agent is in a sodium citrate, sucrose, arginine, polysorbate 80, and has a pH of 6.0, and optionally, the concentration of the anti-TIM-3 agent in the pharmaceutical composition is about 50 mg / mL. In certain embodiments, the anti-LAG-3 active agent is administered in a pharmaceutical composition comprising sodium acetate, trehalose, polysorbate 80, has a pH of 5.5, and optionally, the concentration of the anti-LAG-3 agent in the pharmaceutical composition is about 50 mg / mL.

[0030] In certain embodiments, the present disclosure provides a method, combination therapy, use, or kit for the treatment of cancer in a subject. In certain embodiments, the cancer comprises a tumor. In certain embodiments, the cancer comprises a locally advanced tumor, a metastatic solid tumor, or a combination thereof. In certain embodiments, the cancer comprises a tumor for which a PD-1 inhibitor has been shown.

[0031] In certain embodiments, the subject has received prior treatment with at least one anti-PD-1 / anti-PDL-1 therapy. In certain embodiments, the subject has cancer that has failed prior PD-1 / PDL-1 inhibitor therapy. In certain embodiments, the subject has cancer that progressed during prior PD-1 / PDL-1 inhibitor therapy. In certain embodiments, the cancer includes tumors having acquired resistance to anti-PD-1 therapy. In certain embodiments, the cancer includes tumors having innate resistance to anti-PD-1 therapy. In certain embodiments, the cancer includes tumors having resistance to anti-PD-1 therapy and acquired resistance to anti-PD-1 therapy. In certain embodiments, the tumor has a minimum LAG-3 expression of 5% or more of LAG-3 positive immune cells (e.g., lymphocytes and macrophages) compared to all nucleated cells within the tumor region, as shown by an appropriate immunohistochemistry assay. The tumor region includes tumor cells, intratumoral stroma, and peritumoral stroma and does not include normal and / or adjacent uninvolved tissue. Appropriate LAG-3 immunohistochemistry assays are found in Wojcik et al., Consistent Measurement of LAG-3 expression Across Multiple Staining Platforms with the 17B4 Antibody Clone, bioRxiv (February 22, 2022), doi.org / 10.1101 / 2022.02.21.481075, and Johnson et al., Development of a LAG-3 Immunohistochemistry Assay for Melanoma, bioRxiv (February 26, 2022), doi.org / 10.1101 / 2022.02.25.481964.

[0032] In certain embodiments, the cancer is melanoma. In certain embodiments, the cancer is unresectable and / or metastatic melanoma. In certain embodiments, the cancer is squamous cell carcinoma of the head and neck (SCCHN). In certain embodiments, the cancer is recurrent or metastatic PD-L1+ SCCHN. In certain embodiments, the cancer is endometrial cancer. In certain embodiments, the cancer is MSI-H advanced endometrial cancer. In certain embodiments, the cancer is metastatic endometrial cancer. In certain embodiments, the cancer is endometrial cancer having evidence of disease progression before and after platinum-based chemotherapy and prior PD-L1 inhibitory therapy.

[0033] In certain embodiments, the administration or treatment of the present disclosure produces at least one therapeutic effect. In certain embodiments, the therapeutic effect is a decrease in tumor size. In certain embodiments, the therapeutic effect is a decrease in the number of metastatic lesions over time. In certain embodiments, the therapeutic effect is complete response (e.g., disappearance of all target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is partial response (e.g., a decrease of ≧30% in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is stable disease (e.g., a decrease of ≦30% and an increase of ≦20% in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is progressive disease (e.g., an increase of 20% or more in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is evaluated according to RECIST v1.1 criteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0034]

Figure 1

[0035] I. Active Agents and Compositions Antigen-Binding Proteins and Binding Domains In certain embodiments, the active agents of the present disclosure (e.g., anti-PD-1 active agent, anti-TIM-3 active agent, anti-LAG-3 active agent) include antigen-binding proteins such as antibodies or antigen-binding fragments of antibodies. In certain embodiments, the active agents of the present disclosure (e.g., anti-PD-1 active agent, anti-TIM-3 active agent, anti-LAG-3 active agent) include antibodies or antigen-binding fragments of antibodies.

[0036] In certain embodiments, the antibodies of the present disclosure are monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain molecules and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain - antibody heavy chain pairs, intracellular antibodies, heteroconjugate antibodies, antibody - drug conjugates, single domain antibodies, monovalent antibodies, single chain antibodies or single chain Fv (scFv), camelized antibodies, affibodies, Fab fragments, F(ab’)2 fragments, disulfide - linked Fv (sdFv), anti - idiotypic (anti - Id) antibodies (e.g., anti - anti - Id antibodies), and antigen - binding fragments of any of the above.

[0037] In certain embodiments, the antibody or antibody fragment includes an immunoglobulin molecule or an immunoreactive fragment of an immunoglobulin molecule, i.e., a molecule containing an antigen - binding site. The immunoglobulin molecule can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass (e.g., IgG2a or IgG2b). In certain embodiments, the antibody or antibody fragment is used as a therapeutic agent.

[0038] In certain embodiments, the antibodies of the present disclosure include monoclonal antibodies (mAbs). Monoclonal antibodies generally refer to homogeneous antibodies designed to be directed against a single epitope target or antigenic site, which are generally produced from the immortalization and singular cloning of plasma B cells. In certain embodiments, the monoclonal antibodies and monoclonal antibody variants of the present disclosure include intact monoclonal antibodies, full-length monoclonal antibodies, antibody fragments (such as Fab, Fab’), single-chain variants (scFv), mutant antibodies, fusion proteins containing a portion of a monoclonal antibody, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified configurations of immunoglobulin molecules that include the ability to bind to an antigen recognition site and a target antigen.

[0039] In certain embodiments, the antibody or fragment thereof is produced or sourced by any method known to those of skill in the art (e.g., hybridoma, phage selection, recombinant expression, transgenic animals). In certain embodiments, the antibody is developed in a mouse, rat, or rabbit. In certain embodiments, the antibody is produced by immunizing an animal with an immunogenic collection of cells, cell extracts, or protein preparations containing the desired epitope. Examples of immunogens include, but are not limited to, primary cells, cultured cell lines, cancer cells, proteins, peptides, nucleic acids, or tissues. In certain embodiments, the immunogenic cells can be used as immunogens by themselves or in combination with a non-denaturing adjuvant.

[0040] In certain embodiments, the antibody or fragment thereof is produced by sequencing an existing antibody (or an equivalent antibody fragment or variant that targets the desired epitope) and then recombinantly producing the antibody by any means known in the art. In certain embodiments, the antibody is sequenced and then the polynucleotide sequence is cloned into a vector for expression or propagation. In certain embodiments, the sequence encoding the antibody or antibody fragment of interest is maintained in a vector in a host cell, where the host cell can be grown, frozen, and thawed as needed for future production and use.

[0041] Antibodies (such as IgG antibodies) generally are composed as tetramers having two light chains complexed with two heavy chains. Each light chain contains a light chain variable domain (VL) and a light chain constant domain (CL). Each heavy chain contains a heavy chain variable domain (VH), three heavy chain constant domains (CH1, CH2, and CH3), and a hinge domain located between the CH1 and CH2 domains. The amino-terminal (“N-terminal”) portion of each chain contains the variable domain of amino acids (i.e., the variable region) that is primarily involved in antigen recognition. The carboxy-terminal (“C-terminal”) portion of each chain defines the constant region, with the light chain having a single constant domain and the heavy chain generally having three constant domains and a hinge domain. Generally, the structure of the light chain of an IgG molecule is (N-terminal)-VL-CL-(C-terminal). The light chain variable domain (VL) generally consists of 90 to 115 N-terminal amino acids in the mature light chain sequence. Generally, the structure of the heavy chain of an IgG molecule is (N-terminal)-VH-CH1-hinge-CH2-CH3-(C-terminal). The heavy chain variable domain (VH) generally consists of 110 to 125 N-terminal amino acids in the mature heavy chain sequence.

[0042] The variable domains of antibody molecules generally contain (i) complementarity-determining regions (CDRs) that target and contact residues of an epitope, and (ii) non-CDR framework segments (FRs) that maintain the structure of the domain and determine the positioning of the CDR loops (i.e., enable the CDR loops to contact the target). The VL domain generally has the structure (N-terminus)-FR1-CDRL1-FR2-CDRL2-FR3-CDRL3-FR4-(C-terminus). The VH domain generally has the structure (N-terminus)-FR1-CDRH1-FR2-CDRH2-FR3-CDRH3-FR4-(C-terminus). The terms CDRL1 domain, CDRL2 domain, CDRL3 domain, CDRH1 domain, CDRH2 domain, and CDRH3 domain generally refer to polypeptides that can be incorporated into a protein (including antibodies, antibody fragments, or antibody variants such as single-chain binding molecules or other types of proteins) that binds to an epitope. In certain embodiments, the antibody or fragment thereof comprises an epitope-binding site. In certain embodiments, the epitope-binding site may contain one, two, three, four, five, or all six of the CDR domains of such an antibody.

[0043] In certain embodiments, the antibody or fragment thereof is humanized (i.e., retains the sequence of the antigen-binding portion of a non-human antibody and mutates the non-human remainder of the antibody with human antibody sequences). Generally, a humanized antibody refers to a molecule having (i) an antigen-binding site of an immunoglobulin from a non-human species and (ii) a residual immunoglobulin structure based on the structure and / or sequence of a human immunoglobulin (generally prepared using recombinant techniques known in the art). Without being bound by theory, the general principle of humanizing an antibody involves retaining the basic sequence of the antigen-binding portion of the antibody and then replacing the remaining portion of the non-human antibody with human antibody sequences. General procedures for humanizing monoclonal antibodies are well known in the art, including U.S. Patent Nos. 4,816,567, 5,807,715, 5,866,692, and 6,331,415. In certain embodiments, the humanized antibody retains all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs from a mouse antibody). In certain embodiments, the humanized antibody has one or more (one, two, three, four, five, or six) CDRs that differ in sequence compared to the original antibody.

[0044] Programmed Death-1 (PD-1) Active Agent Programmed death-1 protein (PD-1, also known as PD1 or CD279) is a ~31kD type I membrane protein member of the extended CD28 / CTLA-4 family of T cell regulators. PD-1 is expressed on activated T cells, B cells, monocytes, and (at low levels) natural killer (NK) T cells. The extracellular region of PD-1 contains a single immunoglobulin (Ig) V domain (having approximately 23% identity with the equivalent domain of CTLA-4), followed by a transmembrane region and an intracellular tail containing two phosphorylation sites located in an immunoreceptor tyrosine-based inhibitory motif and an immunoreceptor tyrosine-based switch motif.

[0045] PD-1 has been shown to broadly negatively regulate immune responses, including mediating immune system inhibition by binding to B7-H1 and B7-DC (see, e.g., U.S. Patent Application Publication No. 2005 / 0059051, U.S. Patent Application Publication No. 2007 / 0202100, U.S. Patent Application Publication No. 2008 / 0311117, U.S. Patent Application Publication No. 2009 / 00110667, U.S. Patent Application Publication No. 2009 / 0274666, U.S. Patent Application Publication No. 2009 / 0313687, U.S. Patent Application Publication No. 2009 / 0055944, U.S. Patent No. 10,577,422). The roles of B7-H1 and PD-1 in inhibiting T cell activation and proliferation have led to research on the roles these biomolecules can play as therapeutic targets for the treatment of infection, inflammation, tumors, and cancer (see, e.g., U.S. Patent Application Publication No. 2004 / 0241745, U.S. Patent Application Publication No. 2008 / 0311117, U.S. Patent Application Publication No. 2009 / 0217401, U.S. Patent Application Publication No. 2010 / 0028330, U.S. Patent Application Publication No. 2010 / 0040614, U.S. Patent No. 10,577,422). Antibodies capable of binding to PD-1 have been studied and tested for their therapeutic effects in the treatment of these indications. PD-1 targeting agents such as retilfanlimab have also been shown to effectively maintain or restore the effector functions of PD-1 expressing cells, including T cells, by blocking checkpoint inhibitory interactions between PD-1 and its two major ligands, PD-L1 and PD-L2. Retilfanlimab has also been shown to disrupt the PD-1 / PD-L1 inhibitory axis, enhance IFN-γ secretion in Staphylococcus enterotoxin B (SEB)-stimulated human peripheral blood mononuclear cells, and have activity equivalent to that of replicas of pembrolizumab and nivolumab.

[0046] More than 30 clinical trials have been initiated or completed to study combination therapies that include anti-PD-1 antibodies and anti-PD-1 agents. However, despite the ongoing research on PD-1-based compositions and therapies, there remains a need for improved compositions and treatment regimens that can effectively antagonize and / or block PD-1 / PDL-1 activation, proliferation, and inhibition of T cell activation, which can provide improved therapeutic effects to patients suffering from cancer or the corresponding diseases and conditions.

[0047] The present disclosure provides methods, combination therapies, uses, and kits comprising a therapeutically effective amount of an anti-PD-1 active agent, a therapeutically effective amount of an anti-TIM-3 active agent, and a therapeutically effective amount of an anti-LAG-3 active agent.

[0048] In certain embodiments, the methods, combination therapies, uses, or kits of the present disclosure comprise an anti-PD-1 active agent. In certain embodiments, the anti-PD-1 active agent comprises an anti-PD-1 antibody or a PD-1 binding fragment thereof.

[0049] In certain embodiments, the anti-PD-1 active agent can bind to a continuous or discontinuous (e.g., conformational) portion (epitope) of human PD-1 (i.e., CD279). In certain embodiments, the anti-PD-1 active agent binds to PD-1 molecules of one or more non-human species, including primate species such as cynomolgus monkeys. A representative human PD-1 polypeptide (NCBI sequence NP_005009.2) is as follows: MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL (SEQ ID NO: 31).

[0050] In certain embodiments, the anti-PD-1 antibody is INCMGA00012 (i.e., retifanlimab, MGA012).

[0051] In certain embodiments, the anti-PD-1 agent comprises an anti-PD-1 antibody, a PD-1 binding molecule, or a PD-1 binding fragment described in U.S. Patent No. 10,577,422, which is incorporated herein by reference in connection with anti-PD-1 agents, anti-PD-1 antibodies, PD-1 binding molecules, and PD-1 binding fragments for use in the treatment of cancer, and their corresponding methods, uses, pharmaceutical compositions, treatments, and production methods.

[0052] In certain embodiments, the anti-PD-1 antibody is nivolumab, also known as BMS-936558 or MDX1106 (Bristol-Myers Squibb). In certain embodiments, the anti-PD-1 antibody is pembrolizumab, also known as lambrolizumab or MK-3475 (Merck & Co). In certain embodiments, the anti-PD-1 antibody is pidilizumab, also known as CT-011 (CureTech). In certain embodiments, the anti-PD-1 antibody is MEDI0680, also known as AMP-514 (Medimmune). In certain embodiments, the anti-PD-1 antibody is PDR001 (Novartis). In certain embodiments, the anti-PD-1 antibody is REGN2810 (Regeneron). In certain embodiments, the anti-PD-1 antibody is PF-06801591 (Pfizer). In certain embodiments, the anti-PD-1 antibody is BGB-A317 (BeiGene). In certain embodiments, the anti-PD-1 antibody is TSR-042 (AnaptysBio). In certain embodiments, the anti-PD-1 antibody is SHR-1210 (Hengrui).

[0053] In certain embodiments, the anti-PD-1 active agent comprises a PD-1 binding fragment of an anti-PD-1 antibody, including, but not limited to, immune complexes, diabodies, BiTEs, bispecific antibodies, antibody binding fragments, and similar molecules. In certain embodiments, the anti-PD-1 active agent is as described in U.S. Patent No. 6,808,710, U.S. Patent No. 7,332,582, U.S. Patent No. 7,488,802, U.S. Patent No. 8,008,449, U.S. Patent No. 8,114,845, U.S. Patent No. 8,168,757, U.S. Patent No. 8,354,509, U.S. Patent No. 8,686,119, U.S. Patent No. 8,735,553, U.S. Patent No. 8,747,847, U.S. Patent No. 8,779,105, U.S. Patent No. 8,927,697, U.S. Patent No. 8,920,075, U.S. Patent No. 8,993,731, U.S. Patent No. 9,102,727, U.S. Patent No. 9,205,148, U.S. Patent No. 9,815,897, U.S. Patent No. 9,982,053, U.S. Patent No. 10,414,821, U.S. Patent No. 10,066,013, U.S. Patent No. 10,077,305, U.S. Patent No. 10,160,806, U.S. Patent Application Publication No. 2013 / 0202623 A1, U.S. Patent Application Publication No. 2013 / 0291136 A1, U.S. Patent Application Publication No. 2014 / 0044738 A1, U.S. Patent Application Publication No. 2014 / 0356363 A1, U.S. Patent Application Publication No. 2016 / 0145355 A1, U.S. Patent Application Publication No. 2016 / 0075783 A1, and U.S. Patent Application Publication No. 2019 / 0309069 A1, each of which is incorporated herein by reference as related to anti-PD-1 active agents for use in the treatment of diseases such as cancer.

[0054] In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, or IgG4 isotype. In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises an Fc region of the IgG4 isotype. In certain embodiments, the antibody comprises a hinge domain of the IgG4 isotype that contains a stabilizing mutation. In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises (A) one or more amino acid modifications that reduce the affinity of the variant Fc region for FcγR, and the one or more modifications that reduce the affinity of the variant Fc region for FcγR include the substitutions L234A, L235A, or L234A+L235A, where the numbering is that of the EU index as in Kabat, and / or (B) one or more amino acid modifications that enhance the serum half-life of the variant Fc region, and the one or more modifications that enhance the serum half-life of the variant Fc region include the substitutions M252Y, M252Y+S254T, M252Y+T256E, M252Y+S254T+T256E, or K288D+H435K, where the numbering is that of the EU index in Kabat.

[0055] In certain embodiments, the anti-PD-1 agent is characterized by one or more of the following criteria: (1) binds to human PD-1 endogenously expressed on the surface of activated human T cells; (2) binds to human PD-1 with an equilibrium binding constant (K D ) of 40 nM or less; (3) binds to human PD-1 with an equilibrium binding constant (K D ) of 5 nM or less; (4) binds to human PD-1 with a -1 1.5×104 M -1 min -1 or greater; (5) binds to human PD-1 with an on-rate (k -1 ) of 90.0×104 M a min 4 or greater; (6) binds to human PD-1 with an off-rate (kd) of 7×10 -1 min 4 or less; (7) binds to human PD-1 with an off-rate (k -1 ) of 2×10 d) binds to human PD-1; (8) binds to non-human primate PD-1 (e.g., cynomolgus monkey PD-1), (9) inhibits the binding of PD-1 ligand (PDL-1 / PDL-2) to PD-1 (i.e., blocks or interferes with inhibitory activity), (10) stimulates an immune response, and / or (11) synergizes with an anti-human LAG-3 antibody and / or an anti-human TIM-3 antibody to stimulate an antigen-specific T cell response.

[0056] In certain embodiments, the anti-PD-1 active agent is administered intravenously. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every three weeks or once every four weeks. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every three weeks. In certain embodiments, the anti-PD-1 active agent is administered intravenously once every four weeks.

[0057] PD-1 CDR In certain embodiments, the anti-PD-1 active agent comprises an anti-PD-1 antibody comprising (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain, or a PD-1 binding fragment thereof.

[0058] In certain embodiments, the CDRH1 domain has the amino acid sequence SYWMN (SEQ ID NO: 1).

[0059] In certain embodiments, the CDRH2 domain has the amino acid sequence VIHPSDSETWLDQKFK (SEQ ID NO: 2).

[0060] In certain embodiments, the CDRH3 domain has the amino acid sequence EHYGTSPFAY (SEQ ID NO: 3).

[0061] In certain embodiments, the CDRL1 domain has the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 4).

[0062] In certain embodiments, the CDRL2 domain has the amino acid sequence AASNQGS (SEQ ID NO: 5). In certain embodiments, the CDRL2 domain has the amino acid sequence AASNRGS (SEQ ID NO: 32).

[0063] In certain embodiments, the CDRL3 domain has the amino acid sequence QQSKEVPYT (SEQ ID NO: 6).

[0064] PD-1 VH and VL In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLQQPGAELVRPGASVKLSCKASGYSFTSYWMNWVKQRPGQGLEWIGVIHPSDSETWLDQKFKDKATLTVDKSSTTAYMQLISPTSEDSAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 33). In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to gaggtccaactgcagcagcctggggctgaactggtgaggcctggagcttcagtgaagctgtcctgcaaggcttctggctactccttcaccagctactggatgaactgggtgaagcagaggcctggacaaggccttgagtggattggcgtgattcatccttccgatagtgaaacttggttagatcagaagttcaaggacaaggccacattgactgtagacaaatcctccaccacagcctacatgcaactcatcagcccgacatctgaggactctgcggtctattactgtgcaagggagcactacggtactagcccgtttgcttactggggccaagggactctggtcactgtgtcttcc (SEQ ID NO: 34).

[0065] In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or fragment thereof comprises a humanized amino acid sequence (e.g., hPD-1 mAb).

[0066] In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 7). In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to caagttcaattggtacagagcggggcagaggtgaagaaacccggcgccagtgttaaggtgtcctgcaaagccagcggttacagctttacaagctattggatgaattgggtgcgtcaagcaccagggcagggtctggaatggattggggtgatacatccttctgacagcgaaacatggttggaccagaaatttaaagatcgtgtgacaattacagtcgataagtccacaagcactgcttacatggaactctccagcttgcggtccgaggacaccgctgtgtattattgcgccagagagcactacggcacatcaccttttgcatactggggccagggaactctcgtaaccgtatcctcc (SEQ ID NO: 35).

[0067] In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or a fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWAGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 36). In certain embodiments, the heavy chain variable domain (VH) of the anti-PD-1 antibody or a fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to caagttcaattggtacagagcggggcagaggtgaagaaacccggcgccagtgttaaggtgtcctgcaaagccagcggttacagctttacaagctattggatgaattgggtgcgtcaagcaccagggcagggtctggaatgggctggggtgatacatccttctgacagcgaaacatggttggaccagaaatttaaagatcgtgtgacaattacagtcgataagtccacaagcactgcttacatggaactctccagcttgcggtccgaggacaccgctgtgtattattgcgccagagagcactacggcacatcaccttttgcatactggggccagggaactctcgtaaccgtatcctcc (SEQ ID NO: 37).

[0068] In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to DIVLTQSPASLAVSLGQRATISCRANESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPARFSGSGFGTDFSLNIHPMEEDDAAMYFCQQSKEVPYTFGGGTKLEIK (SEQ ID NO: 38). In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to gacattgtgctgacccaatctccagcttctttggctgtgtctctagggcagagggccaccatctcctgcagagccaacgaaagtgttgataattatggcatgagttttatgaactggttccaacagaaaccaggacagccacccaaactcctcatccatgctgcatccaaccaaggatccggggtccctgccaggtttagtggcagtgggtttgggacagacttcagcctcaacatccatcctatggaggaggatgatgctgcaatgtatttctgtcagcaaagtaaggaggttccgtacacgttcggaggggggaccaagctggaaataaaa (SEQ ID NO: 39).

[0069] In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof comprises a humanized amino acid sequence (e.g., hPD-1 mAb).

[0070] In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIK (SEQ ID NO: 8). In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to gaaatcgtactcacccagtcacctgcaaccctttctctgagccccggtgaacgtgccactctcagctgcagagcaagtgagagtgtggacaattacggcatgtccttcatgaactggtttcagcagaagcctgggcagccacctaagctgctcatccacgccgcctctaaccagggatctggggtgccttcacgtttttctggatcaggaagtggcactgacttcacccttacaatcagctctctggagccagaggactttgccgtctatttctgccagcaatctaaagaggtgccctatacttttggtggcgggaccaaggttgagatcaaa (SEQ ID NO: 40).

[0071] In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRANESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIK (SEQ ID NO: 41). In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to gaaatcgtactcacccagtcacctgcaaccctttctctgagccccggtgaacgtgccactctcagctgcagagcaaatgagagtgtggacaattacggcatgtccttcatgaactggtttcagcagaagcctgggcagccacctaagctgctcatccacgccgcctctaaccagggatctggggtgccttcacgtttttctggatcaggaagtggcactgacttcacccttacaatcagctctctggagccagaggactttgccgtctatttctgccagcaatctaaagaggtgccctatacttttggtggcgggaccaaggttgagatcaaa (SEQ ID NO: 42).

[0072] In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNRGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIK (SEQ ID NO: 43). In certain embodiments, the light chain variable domain (VL) of the anti-PD-1 antibody or fragment thereof is encoded by a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to gaaatcgtactcacccagtcacctgcaaccctttctctgagccccggtgaacgtgccactctcagctgcagagcaagtgagagtgtggacaattacggcatgtccttcatgaactggtttcagcagaagcctgggcagccacctaagctgctcatccacgccgcctctaaccgcggatctggggtgccttcacgtttttctggatcaggaagtggcactgacttcacccttacaatcagctctctggagccagaggactttgccgtctatttctgccagcaatctaaagaggtgccctatacttttggtggcgggaccaaggttgagatcaaa (SEQ ID NO: 44).

[0073] PD-1 HC and LC In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to: QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYCAREHYGTSPFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 9).

[0074] In certain embodiments, the anti-PD-1 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to: EIVLTQSPATLSLSPGERATLSCRASESVDNYGMSFMNWFQQKPGQPPKLLIHAASNQGSGVPSRFSGSGSGTDFTLTISSLEPEDFAVYFCQQSKEVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0075] T Cell Immunoglobulin and Mucin Domain-3 (TIM-3) Active Agent T cell immunoglobulin and mucin domain-3 (TIM-3, also known as TIM3) is a type I membrane protein in the immunoglobulin (Ig) superfamily. TIM-3 is expressed on activated T helper 1 (Th1) and CD8+ T (Tc1) lymphocytes, macrophages, activated natural killer (NK) cells, and IL-17-producing Th17 cells. The extracellular region of TIM-3 contains an extracellular Ig variable-like (IgV) domain, an extracellular mucin-like domain, and a cytoplasmic domain with six conserved tyrosine residues.

[0076] TIM-3 has been shown to function in the inhibition of T cell-, myeloid cell-, and NK cell-mediated responses, as well as in the promotion of immune tolerance. For example, the binding of a neutralizing immunoglobulin domain to the TIM-3 IgV peptide ligand caused Th1 cell hyperproliferation and Th1 cytokine release in immunized mice. Upregulation of TIM-3 expression in CD8+ T cells has also been found in cancer patients. For example, approximately 30% of NY-ESO-1-specific CD8+ T cells in patients with advanced melanoma showed upregulation of TIM-3 expression (see Fourcade et al., J Exp Med 207:2175-86 (2010)).

[0077] The correlation of TIM-3 with T cell-related activity and proliferation has led to research on the role these biomolecules can play as therapeutic targets for the treatment of related diseases including cancer. Specifically, antibodies that can bind to TIM-3 are being studied and tested for their therapeutic efficacy in the treatment of these indications. However, despite ongoing research on TIM-3-based compositions and therapies, there remains a need for improved compositions and treatment regimens that can effectively antagonize and / or block TIM-3 activation, proliferation, and regulation of T cell activation, and provide improved therapeutic effects to patients suffering from cancer or the corresponding diseases and conditions.

[0078] The present disclosure provides methods, combination therapies, uses, and kits comprising a therapeutically effective amount of an anti-PD-1 activating agent, a therapeutically effective amount of an anti-TIM-3 activating agent, and a therapeutically effective amount of an anti-LAG-3 activating agent.

[0079] In certain embodiments, the methods, combination therapies, uses, or kits of the present disclosure comprise an anti-TIM-3 activating agent. In certain embodiments, the anti-TIM-3 activating agent comprises an anti-TIM-3 antibody or a TIM-3 binding fragment thereof.

[0080] In certain embodiments, the anti-TIM-3 activating agent can bind to a continuous or discontinuous (e.g., conformational) portion (epitope) of human TIM-3. In certain embodiments, the anti-TIM-3 activating agent binds to TIM-3 molecules of one or more non-human species, including primate species such as cynomolgus monkeys.

[0081] A representative human TIM-3 polypeptide (Swiss-Prot accession number Q8TDQ0-1) is as follows. SEVEYRAEVGQNAYLPCFYTPAAPGNLVPVCWGKGACPVFECGNVVLRTDERDVNYWTSRYWLNGDFRKGDVSLTIENVTLADSGIYCCRIQIPGIMNDEKFNLKLVIKPAKVTPAPTRQRDFTAAFPRMLTTRGHGPAETQTLGSLPDINLTQISTLANELRDSRLANDLRDSGATIRIGIYIGAGICAGLALALIFGALIFKWYSHSKEKIQNLSLISLANLPPSGLANAVAEGIRSEENIYTIEENVYEVEEPNEYYCYVSSRQQPSQPLGCRFAMP (SEQ ID NO: 45).

[0082] In certain embodiments, the anti-TIM-3 antibody is Antibody A. As used herein, Antibody A refers to an antibody comprising the following heavy chain (HC) and light chain (LC) sequences. The heavy chain has the following amino acid sequence. EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 19), and the light chain has the following amino acid sequence. EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).

[0083] In certain embodiments, the anti-TIM-3 active agent comprises an anti-TIM-3 antibody, TIM-3 binding molecule, or TIM-3 binding fragment described in U.S. Patent No. 10,639,368, which is hereby incorporated by reference in its entirety in connection with anti-TIM-3 active agents, anti-TIM-3 antibodies, TIM-3 binding molecules, and TIM-3 binding fragments for use in the treatment of cancer, as well as their corresponding methods, uses, pharmaceutical compositions, treatments, and methods of production.

[0084] In certain embodiments, the anti-TIM-3 active agent comprises a TIM-3 binding fragment of an anti-TIM-3 antibody, including, but not limited to, immune complexes, diabodies, BiTEs, bispecific antibodies, antibody binding fragments, and similar molecules.

[0085] In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of the IgG1 isotype. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of the IgG1 isotype, and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of the IgG4 isotype. In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises an Fc region of the IgG4 isotype, and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system.

[0086] In certain embodiments, the anti-TIM-3 active agent is administered intravenously. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every two weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every three weeks. In certain embodiments, the anti-TIM-3 active agent is administered intravenously once every four weeks.

[0087] TIM-3 CDR In certain embodiments, the anti-TIM-3 active agent comprises an anti-TIM-3 antibody or a TIM-3 binding fragment thereof comprising (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain.

[0088] In certain embodiments, the CDRH1 domain has the amino acid sequence RQNAWS (SEQ ID NO: 11). In certain embodiments, the CDRH1 domain has the amino acid sequence SSYAMS (SEQ ID NO: 46).

[0089] In certain embodiments, the CDRH2 domain has the amino acid sequence WVSAISGSGGSTY (SEQ ID NO: 12).

[0090] In certain embodiments, the CDRH3 domain has the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 13).

[0091] In certain embodiments, the CDRL1 domain has the amino acid sequence X1ASQSVSSSYLA (SEQ ID NO: 47, wherein X1 is R or G). In certain embodiments, the CDRL1 domain has the amino acid sequence X1ASQSVSSYLA (SEQ ID NO: 48), where X1 is R or G. In certain embodiments, the CDRL1 domain has the amino acid sequence RASQSVSSYLA (SEQ ID NO: 14). In certain embodiments, the CDRL1 domain has the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 49).

[0092] In certain embodiments, the CDRL2 domain has the amino acid sequence X1ASX2RAT (SEQ ID NO: 50), where X1 is D or G and X2 is N, S, or T. In certain embodiments, the CDRL2 domain has the amino acid sequence DASNRAT (SEQ ID NO: 15). In certain embodiments, the CDRL2 domain has the amino acid sequence DASSRAT (SEQ ID NO: 51).

[0093] In certain embodiments, the CDRL3 domain has the amino acid sequence QQYGSSPJT (SEQ ID NO: 52), where J is L or I. In certain embodiments, the CDRL3 domain has the amino acid sequence QQYGSSPLT (SEQ ID NO: 16). In certain embodiments, the CDRL3 domain has the amino acid sequence QQYGSSPIT (SEQ ID NO: 53).

[0094] TIM-3 VH and VL In certain embodiments, the heavy chain variable domain (VH) of the anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSS (SEQ ID NO: 17).

[0095] In certain embodiments, the light chain variable domain (VL) of an anti-TIM-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK (SEQ ID NO: 18).

[0096] TIM-3 HC and LC In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 19).

[0097] In certain embodiments, the anti-TIM-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPASFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).

[0098] Lymphocyte Activation Gene 3 (LAG-3) Active Agent Lymphocyte activation gene 3 (LAG-3, also known as LAG3 and CD223) is a type I membrane protein of the immunoglobulin (Ig) superfamily. LAG-3 is expressed on activated effector T lymphocytes (Teff), activated regulatory T lymphocytes (Treg), activated B lymphocytes, resting natural killer (NK) cells, and resting plasmacytoid dendritic cells (PDC). The extracellular region of LAG-3 contains four extracellular Ig domains and a cytoplasmic domain containing a conserved repeat EP motif and a single conserved KIEELE motif.

[0099] LAG-3 has been shown to function in the negative regulation of activated T cells, particularly by targeting MHC class II ligands expressed on antigen-presenting cells (APC) and activated T cells. The interaction between LAG-3 and MHC class II has been shown to inhibit the proliferation and cytokine secretion of CD4+ and CD8+ Teff cells.

[0100] The correlation between the immune response of LAG-3 and the corresponding regulation of T cell-related activities and proliferation has led to research on the role that these biomolecules can function as therapeutic targets for the treatment of related diseases including cancer. Specifically, antibodies that can bind to LAG-3 have been studied and validated for their therapeutic effects in the treatment of these indications. However, despite the ongoing research on LAG-3-based compositions and therapies, there remains a need for improved compositions and treatment regimens that can effectively antagonize and / or block LAG-3 activation, proliferation, and regulation of T cell activation, which can provide improved therapeutic effects to patients suffering from cancer or the corresponding diseases and conditions.

[0101] The present disclosure provides methods, combination therapies, uses, and kits comprising a therapeutically effective amount of an anti-PD-1 active agent, a therapeutically effective amount of an anti-TIM-3 active agent, and a therapeutically effective amount of an anti-LAG-3 active agent.

[0102] In certain embodiments, the methods, combination therapies, uses, or kits of the present disclosure comprise an anti-LAG-3 active agent. In certain embodiments, the anti-LAG-3 active agent comprises an anti-LAG-3 antibody or a LAG-3 binding fragment thereof.

[0103] In certain embodiments, the anti-LAG-3 active agent can bind to a continuous or discontinuous (e.g., conformational) portion (epitope) of human LAG-3. In certain embodiments, the anti-LAG-3 active agent binds to LAG-3 molecules of one or more non-human species, including primate species such as cynomolgus monkeys.

[0104] The representative human LAG-3 polypeptide (GenBank accession number NM_002286.5) is known as VPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLILGVLSLLLLVTGAFGFHLWRRQWRPRRFSALEQGIHPPQAQSKIEELEQEPEPEPEPEPEPEPEPEPEQL (SEQ ID NO: 54).

[0105] In certain embodiments, the anti-LAG-3 antibody is Antibody B. As used herein, Antibody B describes an antibody comprising the following heavy chain (HC) and light chain (LC) sequences. The heavy chain has the following amino acid sequence: QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 29). The light chain has the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).

[0106] In certain embodiments, the anti-LAG-3 active agent comprises an anti-LAG-3 antibody, a LAG-LAG-3 binding molecule, or a LAG-3 binding fragment as described in U.S. Patent No. 10,844,119, which is hereby incorporated by reference in its entirety for anti-LAG-3 active agents, anti-LAG-3 antibodies, LAG-3 binding molecules, and LAG-3 binding fragments for use in the treatment of cancer, and their corresponding methods, uses, pharmaceutical compositions, treatments, and methods of production.

[0107] In certain embodiments, the anti-LAG-3 active agent comprises a LAG-3 binding fragment of an anti-LAG-3 antibody, including (but not limited to) the same molecules including immune complexes, diabodies, BiTEs, bispecific antibodies, antibody binding fragments, and similar molecules.

[0108] In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG1 isotype. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG1 isotype and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG4 isotype. In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises an Fc region of the IgG4 isotype and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system.

[0109] In certain embodiments, the anti-LAG-3 active agent is administered intravenously. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every two weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every three weeks. In certain embodiments, the anti-LAG-3 active agent is administered intravenously once every four weeks.

[0110] LAG-3 CDR In certain embodiments, the anti-LAG-3 active agent comprises an anti-LAG-3 antibody or a LAG-3 binding fragment thereof, comprising (i) a heavy chain variable domain (VH) comprising a CDRH1 domain, a CDRH2 domain, and a CDRH3 domain, and (ii) a light chain variable domain (VL) comprising a CDRL1 domain, a CDRL2 domain, and a CDRL3 domain.

[0111] In certain embodiments, the CDRH1 domain has the amino acid sequence DTYIH (SEQ ID NO: 21).

[0112] In certain embodiments, the CDRH2 domain has the amino acid sequence EIDPANDNTKYDPKFQG (SEQ ID NO: 22).

[0113] In certain embodiments, the CDRH3 domain has the amino acid sequence YYYX1YX2VGGFDY (SEQ ID NO: 55), wherein X1 is K or R and X2 is D or E. In certain embodiments, the CDRH3 domain has the amino acid sequence YYYKYDVGGFDY (SEQ ID NO: 23).

[0114] In certain embodiments, the CDRL1 domain has the amino acid sequence SVSSSISSSNLH (SEQ ID NO: 24).

[0115] In certain embodiments, the CDRL2 domain has the amino acid sequence GTSNLAS (SEQ ID NO: 25).

[0116] In certain embodiments, the CDRL3 domain has the amino acid sequence QQWSSYPFT (SEQ ID NO: 26).

[0117] LAG-3 VH and VL In certain embodiments, the heavy chain variable domain (VH) of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSS (SEQ ID NO: 27).

[0118] In certain embodiments, the light chain variable domain (VL) of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIK (SEQ ID NO: 28).

[0119] LAG-3 HC and LC In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRSEDTAVYYCATYYYKYDVGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 29).

[0120] In certain embodiments, the anti-LAG-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).

[0121] II. Administration and Treatment Pharmaceutical Compositions In certain embodiments, the active agents of the present disclosure are formulated as, or included in, a pharmaceutical composition. In certain embodiments, such compositions comprise one or more active agents of the present disclosure (e.g., an anti-PD-1 active agent, an anti-LAG-3 active agent, an anti-TIM-3 active agent), and one or more pharmaceutically acceptable excipients (e.g., a carrier, solvent, or delivery vehicle). In certain embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of an anti-PD-1 active agent. In certain embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of an anti-LAG-3 active agent. In certain embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of an anti-TIM-3 active agent.

[0122] In certain embodiments, the pharmaceutical composition of the present disclosure is an aqueous composition (i.e., a composition containing water). In certain embodiments, the pharmaceutical composition of the present disclosure comprises water, sterile or disinfected water, or water for injection (WFI).

[0123] In certain embodiments, the pharmaceutical composition of the present disclosure comprises one or more of a pH buffer solution (e.g., acetate buffer, phosphate buffered saline (PBS), HEPES, TES, MOPS), isotonic saline, Ringer's solution, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), alginic acid, ethyl alcohol, an emulsifier (e.g., polysorbate such as polysorbate 80), and pharmaceutically acceptable mixtures thereof. In certain embodiments, the pharmaceutical composition of the present disclosure comprises phosphate buffered saline (PBS) or an acetate component (e.g., acetic acid, acetate, and / or acetate buffer). In certain embodiments, the pharmaceutical composition of the present disclosure comprises thickening and solubilizing agents such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0124] In certain embodiments, the pharmaceutical composition of the present disclosure includes a carrier. In certain embodiments, the carrier is one or more sterile liquids such as water and oil. In certain embodiments, the carrier is of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and similar oils. In certain embodiments, the pharmaceutical composition includes physiological saline, aqueous dextrose, and / or glycerol solutions (especially for injection solutions). Other suitable pharmaceutical carriers include, but are not limited to, starch, glucose, lactose, sucrose, trehalose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrin, water, ethanol, and the like. In certain embodiments, the pharmaceutical composition includes a wetting agent or emulsifier, or a pH buffering agent. In some embodiments, the emulsifier includes polysorbate (e.g., polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80).

[0125] In certain embodiments, the pharmaceutical compositions of the present disclosure include an adjuvant. Non-limiting examples of suitable adjuvants include complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA), RIBI adjuvant system (RAS), TiterMax, muramyl peptides, Syntex adjuvant formulation (SAF), alum (aluminum hydroxide and / or aluminum phosphate), aluminum salt adjuvants, Gerbu adjuvant (GERBU Biochemicals GmbH), nitrocellulose-absorbed antigen, encapsulated or entrapped antigen, 3 De-O-acylated monophosphoryl lipid A (3 D-MPL), immunostimulatory oligonucleotides, toll-like receptor (TLR) ligands, mannose-binding lectin (MBL) ligands, stimulator of interferon genes (STING) agonists, saponins, Quil A, QS-21, QS-7, immunostimulatory complexes such as ISCOMATRIX, but are not limited thereto. Other adjuvants include CpG oligonucleotides, as well as double-stranded RNA molecules such as poly(A) and poly(U).

[0126] In certain embodiments, the pharmaceutical compositions of the present disclosure include one or more pharmaceutically acceptable excipients. Excipients include, for example, anti-adhesives, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavoring agents, fragrances, lubricants (glidants), lubricants, preservatives, printing inks, absorbents, suspending or dispersing agents, sweeteners, and water of hydration. Exemplary excipients include acetic acid, aluminum stearate, butylated hydroxytoluene (BHT), calcium carbonate, calcium chloride, calcium phosphate (dibasic), calcium stearate, carboxymethyl cellulose, croscarmellose, crospovidone, cysteine, ethyl cellulose, gelatin, glucose, glucuronic acid, gluconic acid, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxybutanedioic acid, inositol, lactose, magnesium chloride, magnesium stearate, maltitol, mannitol, methionine, methyl cellulose, methylparaben, microcrystalline cellulose, phosphoric acid, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, sucrose, shellac, silicon dioxide, sodium acetate, sodium carbonate, sodium bicarbonate, sodium carboxymethyl cellulose, sodium chloride, sodium citrate, sodium hydroxide, sodium phosphate, sodium glycolate starch, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, xylitol, zinc stearate, and combinations thereof, but are not limited thereto.

[0127] In certain embodiments, the pharmaceutical compositions of the present disclosure include an aqueous vehicle, a non-aqueous vehicle, an antibacterial agent, an isotonic agent, a buffer, an antioxidant, a suspending and dispersing agent, an emulsifying agent, a sequestering or chelating agent, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose, and lactated Ringer's injection. Examples of non-aqueous vehicles include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil, and peanut oil. Examples of antibacterial agents include phenol or cresol, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Examples of isotonic agents include sodium chloride and dextrose. Examples of buffers include acetates, phosphates, citrates, and other organic acids. Examples of antioxidants include tocopherol (e.g., vitamin E), sodium bisulfate, ascorbic acid, and methionine. Examples of suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. An example of an emulsifying agent is polysorbate 80 (TWEEN® 80). An example of a sequestering or chelating agent is EDTA. Other examples of pharmaceutical vehicles include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles, and sodium hydroxide, hydrochloric acid, citric acid, acetic acid, or lactic acid for pH adjustment.

[0128] In certain embodiments, the pharmaceutical compositions of the present disclosure include octadecyl dimethyl benzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol, low molecular weight (less than about 10 residues) polypeptides, proteins, gelatin such as serum albumin, or immunoglobulins, hydrophilic polymers such as polyvinyl pyrrolidone, and amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine, monosaccharides, disaccharides, other carbohydrates including glucose, and chelating agents such as mannitol, or dextrin, EDTA, sugars such as sucrose, mannitol, trehalose or sorbitol, salt-forming counterions such as sodium, metal complexes (e.g., Zn protein complexes), and / or nonionic surfactant agents such as TWEEN™, preservatives such as PLURONIC™ or polyethylene glycol (PEG).

[0129] In certain embodiments, the components of the compositions of the present disclosure are supplied separately or mixed together in unit dosage forms. In certain embodiments where the composition is administered by infusion, it can be dispensed in infusion bottles containing sterile pharmaceutical grade water or saline. In certain embodiments where the composition is administered by injection, ampoules of sterile water for injection or saline can be provided and the components can be mixed prior to administration.

[0130] Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc., but are not limited thereto. In certain embodiments, the pharmaceutical composition takes the form of a solution, suspension, emulsion, powder, subcutaneous delivery formulation, sustained release composition, etc.

[0131] Therapeutic Uses and Administration In certain embodiments, the active agents and compositions of the present disclosure are used as therapeutic agents for preventing and / or treating diseases in a subject (e.g., cancer). In certain embodiments, the active agents and compositions of the present disclosure are used as therapeutic agents for modulating (e.g., negatively or positively modulating) the proliferation, function, and homeostasis of T cells in a subject.

[0132] In certain embodiments, the present disclosure provides a method for treating a disease (e.g., cancer) in a human subject in need thereof. In certain embodiments, the present disclosure provides a method for doing so in a human subject in need of modulating (e.g., negatively or positively modulating) T cell proliferation, function, and homeostasis. In certain embodiments, the method comprises administering to the subject (i) a therapeutically effective amount of an anti-PD-1 agent (e.g., retifanlimab), (ii) a therapeutically effective amount of an anti-TIM-3 agent (e.g., Antibody A), and (iii) a therapeutically effective amount of an anti-LAG-3 agent (e.g., Antibody B). In certain embodiments, the method comprises administering to the subject a combination therapy agent of (i) an anti-PD-1 agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; (ii) an anti-TIM-3 agent at about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; and (iii) an anti-LAG-3 agent at about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg.In certain embodiments, the method comprises administering to the subject (i) an anti-PD-1 active agent of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, once every 3 to 5 weeks, and in some embodiments once every 3 weeks or once every 4 weeks, (ii) an anti-TIM-3 active agent of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks, (iii) an anti-LAG-3 active agent of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks.

[0133] In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 500 mg, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 400 mg, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 350 mg. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 375 mg, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 500 mg, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 450 mg. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 375 mg, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 1000 mg, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 750 mg. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 500 mg once every four weeks, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 400 mg once every two weeks, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 350 mg once every two weeks. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 375 mg once every three weeks, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 500 mg once every three weeks, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 450 mg once every three weeks. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 375 mg once every three weeks, (ii) an anti-TIM-3 agent (e.g., antibody A) of about 1000 mg once every three weeks, and (iii) an anti-LAG-3 agent (e.g., antibody B) of about 750 mg once every three weeks.

[0134] In certain embodiments, the method comprises administering to a subject (i) a therapeutically effective amount of an anti-PD-1 agent (e.g., retifanlimab), and (ii) a therapeutically effective amount of an anti-TIM-3 agent (e.g., Antibody A). In certain embodiments, the method comprises administering to the subject a combination therapy agent of (i) an anti-PD-1 agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, and (ii) an anti-TIM-3 agent at about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg. In certain embodiments, the method comprises administering to the subject (i) an anti-PD-1 agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg once every 3 to 5 weeks, and in some embodiments once every 3 weeks or once every 4 weeks, and (ii) an anti-TIM-3 agent at about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks. In certain embodiments, the method comprises administering to the subject (i) an anti-PD-1 agent at about 500 mg (e.g., retifanlimab) once every 4 weeks, and (ii) an anti-TIM-3 agent at about 400 mg (e.g., Antibody A) once every 2 weeks. In certain embodiments, the method comprises administering to the subject (i) an anti-PD-1 agent at about 375 mg (e.g., retifanlimab) once every 3 weeks, and (ii) an anti-TIM-3 agent at about 500 mg (e.g., Antibody A) once every 3 weeks.In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 agent (e.g., retifanlimab) of about 375 mg once every three weeks, and (ii) an anti-TIM-3 agent (e.g., antibody A) of about 1000 mg once every three weeks.

[0135] In certain embodiments, the method comprises administering to a subject (i) a therapeutically effective amount of an anti-PD-1 active agent (e.g., retifanlimab), and (ii) a therapeutically effective amount of an anti-LAG-3 active agent (e.g., Antibody B). In certain embodiments, the method comprises administering to a subject a combination therapy agent of (i) an anti-PD-1 active agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, and (ii) an anti-LAG-3 active agent at about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 active agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, once every 3 to 5 weeks in some embodiments, and once every 3 weeks or once every 4 weeks in some embodiments, and (ii) an anti-LAG-3 active agent at about 300 to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg, once every 1 to 4 weeks, once every 2 weeks, once every 3 weeks, or once every 4 weeks in some embodiments. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 active agent at about 500 mg (e.g., retifanlimab) once every 4 weeks, and (ii) an anti-LAG-3 active agent at about 400 mg (e.g., Antibody B) once every 2 weeks. In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 active agent at about 375 mg (e.g., retifanlimab) once every 3 weeks, and (ii) an anti-LAG-3 active agent at about 450 mg (e.g., Antibody B) once every 3 weeks.In certain embodiments, the method comprises administering to a subject (i) an anti-PD-1 active agent (e.g., retifanlimab) at about 375 mg once every three weeks, and (ii) an anti-LAG-3 active agent (e.g., antibody B) at about 750 mg once every three weeks.

[0136] In certain embodiments, the method comprises administering to a subject (i) a therapeutically effective amount of an anti-TIM-3 active agent (e.g., antibody A), and (ii) a therapeutically effective amount of an anti-LAG-3 active agent (e.g., antibody B). In certain embodiments, the method comprises administering to the subject a combination therapy agent of (i) an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg, and (ii) an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the method comprises administering to the subject (i) an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks, and (ii) an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg once every 1 to 4 weeks, in some embodiments once every 2 weeks, once every 3 weeks, or once every 4 weeks. In certain embodiments, the method comprises administering to the subject (i) an anti-TIM-3 active agent (e.g., antibody A) in an amount of about 400 mg once every 2 weeks, and (ii) an anti-LAG-3 active agent (e.g., antibody B) in an amount of about 350 mg once every 2 weeks. In certain embodiments, the method comprises administering to the subject (i) an anti-TIM-3 active agent (e.g., antibody A) in an amount of about 500 mg once every 3 weeks, and (ii) an anti-LAG-3 active agent (e.g., antibody B) in an amount of about 450 mg once every 3 weeks. In certain embodiments, the method comprises administering to the subject (i) an anti-TIM-3 active agent (e.g., antibody A) in an amount of about 1000 mg once every 3 weeks, and (ii) an anti-LAG-3 active agent (e.g., antibody B) in an amount of about 750 mg once every 3 weeks.

[0137] In certain embodiments, the present disclosure provides a combination therapy comprising a therapeutically effective amount (e.g., from about 300 mg to about 600 mg, in some embodiments from about 375 mg to about 550 mg, in some embodiments from about 375 mg to about 500 mg, in some embodiments from about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg) of an anti-PD-1 agent, a therapeutically effective amount (e.g., from about 300 mg to about 1000 mg, in some embodiments from about 400 mg to about 1000 mg, in some embodiments from about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg) of an anti-TIM-3 agent, and a therapeutically effective amount (e.g., from about 300 mg to about 800 mg, in some embodiments from about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg) of an anti-LAG-3 agent.

[0138] In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 500 mg, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 400 mg, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 350 mg. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 375 mg, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 500 mg, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 450 mg. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 375 mg, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 1000 mg, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 750 mg. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 500 mg formulated for administration once every four weeks, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 400 mg formulated for administration once every two weeks, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 350 mg formulated for administration once every two weeks. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 375 mg formulated for administration once every three weeks, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 500 mg formulated for administration once every three weeks, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 450 mg formulated for administration once every three weeks. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 active agent (e.g., retifanlimab) of about 375 mg formulated for administration once every three weeks, (ii) an anti-TIM-3 active agent (e.g., antibody A) of about 1000 mg formulated for administration once every three weeks, and (iii) an anti-LAG-3 active agent (e.g., antibody B) of about 750 mg formulated for administration once every three weeks.

[0139] In certain embodiments, the combination therapy comprises: (i) a therapeutically effective amount of an anti-PD-1 agent (e.g., retifanlimab), and (ii) a therapeutically effective amount of an anti-TIM-3 agent (e.g., Antibody A). In certain embodiments, the combination therapy comprises administering a combination therapeutic agent of: (i) an anti-PD-1 agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, and (ii) an anti-TIM-3 agent at about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg. In certain embodiments, the combination therapy comprises: (i) an anti-PD-1 agent at about 500 mg (e.g., retifanlimab), and (ii) an anti-TIM-3 agent at about 400 mg (e.g., Antibody A). In certain embodiments, the combination therapy comprises: (i) an anti-PD-1 agent at about 375 mg (e.g., retifanlimab), and (ii) an anti-TIM-3 agent at about 500 mg (e.g., Antibody A). In certain embodiments, the combination therapy comprises: (i) an anti-PD-1 agent at about 375 mg (e.g., retifanlimab), and (ii) an anti-TIM-3 agent at about 1000 mg (e.g., Antibody A).

[0140] In certain embodiments, the combination therapy comprises (i) a therapeutically effective amount of an anti-PD-1 agent (e.g., retifanlimab), and (ii) a therapeutically effective amount of an anti-LAG-3 agent (e.g., antibody B). In certain embodiments, the combination therapy comprises administering a combination therapy agent of (i) an anti-PD-1 agent at about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg, and (ii) an anti-LAG-3 agent at about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the combination therapy comprises (i) an anti-PD-1 agent at about 500 mg (e.g., retifanlimab), and (ii) an anti-LAG-3 agent at about 400 mg (e.g., antibody B). In certain embodiments, the combination therapy comprises (i) an anti-PD-1 agent at about 375 mg (e.g., retifanlimab), and (ii) an anti-LAG-3 agent at about 450 mg (e.g., antibody B). In certain embodiments, the combination therapy comprises (i) an anti-PD-1 agent at about 375 mg (e.g., retifanlimab), and (ii) an anti-LAG-3 agent at about 750 mg (e.g., antibody B).

[0141] In certain embodiments, the combination therapy comprises (i) a therapeutically effective amount of an anti-TIM-3 active agent (e.g., Antibody A) and (ii) a therapeutically effective amount of an anti-LAG-3 active agent (e.g., Antibody B). In certain embodiments, the combination therapy comprises (i) from about 300 mg to about 1000 mg, in some embodiments from about 400 mg to about 1000 mg, in some embodiments from about 350 mg to about 500 mg, and in some embodiments about 400 mg, about 500 mg, or about 1000 mg of the anti-TIM-3 active agent, and (ii) from about 300 mg to about 800 mg, in some embodiments from about 350 mg to about 750 mg, and in some embodiments about 350 mg, about 450 mg, or about 750 mg of the anti-LAG-3 active agent. In certain embodiments, the combination therapy comprises (i) about 400 mg of the anti-TIM-3 active agent (e.g., Antibody A) and (ii) about 350 mg of the anti-LAG-3 active agent (e.g., Antibody B). In certain embodiments, the combination therapy comprises (i) about 500 mg of the anti-TIM-3 active agent (e.g., Antibody A) and (ii) about 450 mg of the anti-LAG-3 active agent (e.g., Antibody B). In certain embodiments, the combination therapy comprises (i) about 1000 mg of the anti-TIM-3 active agent (e.g., Antibody A) and (ii) about 750 mg of the anti-LAG-3 active agent (e.g., Antibody B).

[0142] In certain embodiments, the combination therapy is for use in the treatment of cancer in a human subject in need thereof. In certain embodiments, the combination therapy of the present disclosure is for use in a method for treating a disease in a human subject (e.g., cancer) in need thereof. In certain embodiments, the combination therapy of the present disclosure is for use in performing it in a human subject in need of modulating (e.g., negatively or positively regulating) T cell proliferation, function, and homeostasis. In certain embodiments, the combination therapy of the present disclosure is for use in the manufacture / production of a medicament for use in the treatment of a disease in a human subject (e.g., cancer) in need thereof. In certain embodiments, the combination therapy of the present disclosure is for use in the manufacture / production of a medicament for use in a human subject in need thereof for modulating (e.g., negatively or positively regulating) T cell proliferation, function, and homeostasis.

[0143] In certain embodiments, the present disclosure provides a kit for treating a disease (e.g., cancer) in a human subject in need thereof. In certain embodiments, the present disclosure provides a kit for doing so in a human subject in need of modulating (e.g., negatively or positively regulating) T cell proliferation, function, and homeostasis. In certain embodiments, the kit comprises an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; an anti-TIM-3 active agent in an amount of about 300 to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; and an anti-LAG-3 active agent in an amount of about 300 to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the kit comprises instructions for administering the anti-PD-1 active agent (e.g., once every three weeks or once every four weeks), and instructions for administering the anti-TIM-3 and anti-LAG-3 active agents (e.g., once every two weeks, once every three weeks, or once every four weeks). In certain embodiments, the kit comprises an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg; and an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg.In certain embodiments, the kit includes instructions for administering an anti-PD-1 active agent (e.g., once every three weeks or once every four weeks), and instructions for administering an anti-TIM-3 and anti-LAG-3 active agent (e.g., once every two weeks, once every three weeks, or once every four weeks).

[0144] In certain embodiments, the kit includes (i) an anti-PD-1 active agent of about 500 mg (e.g., retifanlimab), (ii) an anti-TIM-3 active agent of about 400 mg (e.g., antibody A), and (iii) an anti-LAG-3 active agent of about 350 mg (e.g., antibody B), and also includes instructions for administering the anti-PD-1 active agent once every four weeks and the anti-TIM-3 and anti-LAG-3 active agents once every two weeks.

[0145] In certain embodiments, the kit includes (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab), (ii) an anti-TIM-3 active agent of about 500 mg (e.g., antibody A), and (iii) an anti-LAG-3 active agent of about 450 mg (e.g., antibody B), and also includes instructions for administering the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent once every three weeks.

[0146] In certain embodiments, the kit includes (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab), (ii) an anti-TIM-3 active agent of about 1000 mg (e.g., antibody A), and (iii) an anti-LAG-3 active agent of about 750 mg (e.g., antibody B), and also includes instructions for administering the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent once every three weeks.

[0147] In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; and (ii) an anti-TIM-3 active agent in an amount of about 300 mg to about 1000 mg, in some embodiments about 400 mg to about 1000 mg, in some embodiments about 350 mg to about 500 mg, in some embodiments about 400 mg, about 500 mg or about 1000 mg. In certain embodiments, the kit includes instructions for administering the anti-PD-1 active agent (e.g., once every 3 weeks or once every 4 weeks), and instructions for administering the anti-TIM-3 active agent (e.g., once every 2 weeks, once every 3 weeks, or once every 4 weeks). In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 500 mg (e.g., retifanlimab); and (ii) an anti-TIM-3 active agent of about 400 mg (e.g., antibody A), and includes instructions for administering the anti-PD-1 active agent once every 4 weeks and the anti-TIM-3 active agent once every 2 weeks. In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab); and (ii) an anti-TIM-3 active agent of about 500 mg (e.g., antibody A), and includes instructions for administering the anti-PD-1 active agent and the anti-TIM-3 active agent once every 3 weeks. In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab); and (ii) an anti-TIM-3 active agent of about 1000 mg (e.g., antibody A), and includes instructions for administering the anti-PD-1 active agent and the anti-TIM-3 active agent once every 3 weeks.

[0148] In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent in an amount of about 300 mg to about 600 mg, in some embodiments about 375 mg to about 550 mg, in some embodiments about 375 mg to about 500 mg, in some embodiments about 450 mg to about 550 mg, in some embodiments about 375 mg or about 500 mg; and (ii) an anti-LAG-3 active agent in an amount of about 300 mg to about 800 mg, in some embodiments about 350 mg to about 750 mg, in some embodiments about 350 mg, about 450 mg or about 750 mg. In certain embodiments, the kit includes instructions for administering the anti-PD-1 active agent (e.g., once every 3 weeks or once every 4 weeks) and instructions for administering the anti-LAG-3 active agent (e.g., once every 2 weeks, once every 3 weeks, or once every 4 weeks). In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 500 mg (e.g., retifanlimab); and (ii) an anti-LAG-3 active agent of about 350 mg (e.g., Antibody B), and includes instructions for administering the anti-PD-1 active agent once every 4 weeks and the anti-LAG-3 active agent once every 2 weeks. In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab); and (ii) an anti-LAG-3 active agent of about 450 mg (e.g., Antibody B), and includes instructions for administering the anti-PD-1 active agent and the anti-LAG-3 active agent once every 3 weeks. In certain embodiments, the kit comprises: (i) an anti-PD-1 active agent of about 375 mg (e.g., retifanlimab); and (ii) an anti-LAG-3 active agent of about 750 mg (e.g., Antibody B), and also includes instructions for administering the anti-PD-1 active agent and the anti-LAG-3 active agent once every 3 weeks.

[0149] In certain embodiments, the kit comprises (i) an anti-TIM-3 active agent of from about 300 mg to about 1000 mg, in some embodiments from about 400 mg to about 1000 mg, in some embodiments from about 350 mg to about 500 mg, and in some embodiments about 400 mg, about 500 mg, or about 1000 mg, and (ii) an anti-LAG-3 active agent of from about 300 mg to about 800 mg, in some embodiments from about 350 mg to about 750 mg, and in some embodiments about 350 mg, about 450 mg, or about 750 mg. In certain embodiments, the kit comprises instructions for administering the anti-TIM-3 and anti-LAG-3 active agents (e.g., once every two weeks, once every three weeks, or once every four weeks). In certain embodiments, the kit comprises (i) an anti-TIM-3 active agent of about 400 mg (e.g., antibody A), and (ii) an anti-LAG-3 active agent of about 350 mg (e.g., antibody B), and instructions for administering the anti-TIM-3 and anti-LAG-3 active agents once every two weeks. In certain embodiments, the kit comprises (i) an anti-TIM-3 active agent of about 500 mg (e.g., antibody A), and (ii) an anti-LAG-3 active agent of about 450 mg (e.g., antibody B), and instructions for administering the anti-TIM-3 active agent and the anti-LAG-3 active agent once every three weeks. In certain embodiments, the kit comprises (i) an anti-TIM-3 active agent of about 1000 mg (e.g., antibody A), and (ii) an anti-LAG-3 active agent of about 750 mg (e.g., antibody B), and instructions for administering the anti-TIM-3 active agent and the anti-LAG-3 active agent once every three weeks.

[0150] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used for the treatment of cancer.Examples of cancers that can be treated include adrenal tumors, AIDS-related cancers, alveolar soft part sarcoma, anal cancer, squamous cell anal canal cancer (SCAC), astrocytic tumors, bladder cancer, bone cancer, brain and spinal cord cancer, metastatic brain tumors, breast cancer, HER2+ breast cancer, triple-negative breast cancer (TNBC), carotid body tumors, cervical cancer, HPV-related cervical cancer, chondrosarcoma, chondroma, chromophobe renal cell cancer, clear cell cancer, colon cancer, colorectal cancer, cutaneous benign fibrous histiocytoma, histiocytic small round cell tumor, epithelioma, endometrial cancer, metastatic endometrial cancer, unselected endometrial cancer, MSI-high endometrial cancer, dMMR endometrial cancer, DNA polymerase ε (POLE) exonuclease domain mutation-positive endometrial cancer, Ewing tumor, Ewing sarcoma, extraskeletal myxoid chondrosarcoma, osteogenesis imperfecta, fibrous dysplasia of bone, gallbladder cancer or cholangiocarcinoma, cholangiocarcinoma, gastric cancer, gastroesophageal junction (GEJ) cancer, gestational trophoblastic disease, germ cell tumors, glioma, glioblastoma, head and neck cancer, head and neck squamous cell cancer (SCCHN) including recurrent or metastatic PD-L1+ SCCHN, hematologic malignancies, hepatocellular cancer, islet cell tumors, Kaposi sarcoma, kidney cancer, renal cell cancer (RCC), clear cell RRC, papillary RCC, and chromophobe RCC, leukemia, acute myeloid leukemia, lipoma / benign lipomatous tumor, liposarcoma / malignant lipomatous tumor, liver cancer, hepatocellular liver cancer (HCC), lymphoma, diffuse large B-cell lymphoma (DLBCL), non-Hodgkin lymphoma (NHL), lung cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), medulloblastoma, melanoma (unresectable and / or metastatic), uveal melanoma, meningioma, mesothelioma, mesothelial pharyngeal cancer, multiple endocrine neoplasia, multiple myeloma, myelodysplastic syndromes, neuroblastoma, neuroendocrine tumors, ovarian cancer, pancreatic cancer, papillary thyroid cancer, parathyroid tumors, pediatric cancer, peripheral nerve sheath tumors, pharyngeal cancer, chromophobe cell tumor, pituitary tumors, prostate cancer, metastatic castration-resistant prostate cancer (mCRPC), posterior uveal melanoma, rare blood disorders, renal metastatic cancer, rhabdoid tumor, rhabdomyosarcoma, sarcoma, skin cancer, Merkel cell cancer, small round blue cell tumor, neuroblastoma, rhabdomyosarcoma, soft tissue sarcoma, squamous cell cancer, gastric cancer, synovial sarcoma, testicular cancer, thymic cancer, thymoma, thyroid metastatic cancer, urothelial cancer, and uterine cancer, but are not limited to these.

[0151] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used for the treatment of melanoma (unresectable and / or metastatic), colorectal cancer, hepatocellular carcinoma, glioma, kidney cancer, breast cancer, multiple myeloma, bladder cancer, neuroblastoma, sarcoma, non-Hodgkin lymphoma, non-small cell lung cancer, ovarian cancer, pancreatic cancer, and rectal cancer. In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used for the treatment of (unresectable and / or metastatic) melanoma. In certain embodiments, the cancer is melanoma. In certain embodiments, the cancer is unresectable and / or metastatic melanoma.

[0152] In certain embodiments, the cancer being treated is characterized by solid tumors. In certain embodiments, the cancer being treated is characterized by metastatic lesions. In one embodiment, the cancer is a solid tumor. Examples of solid tumors include malignant tumors (e.g., sarcomas and cancers) of various organs such as the lung, breast, ovary, lymph, gastrointestinal tract (e.g., colon), genitourinary tract (e.g., kidney, urinary tract, bladder cells, prostate), pharynx, CNS (e.g., brain, nerve or glial cells), head and neck, skin (e.g., melanoma), pancreas, colon, rectum, renal cell carcinoma, liver, small intestine, and those affecting the esophagus. In certain embodiments, the cancer is early, intermediate, advanced, or metastatic cancer. In certain embodiments, the cancer is associated with an increase in PD-1 activity (e.g., an increase in PD-1 expression).

[0153] In certain embodiments, the cancer is a blood cancer, such as leukemia, lymphoma, or myeloma. In certain embodiments, the cancer is leukemia, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), chronic lymphocytic leukemia (CLL), or hairy cell leukemia. In certain embodiments, the cancer is a lymphoma, such as B cell lymphoma, diffuse large B cell lymphoma (DLBCL), activated B cell-like (ABC) diffuse large B cell lymphoma, germinal center B cell (GCB) diffuse large B cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, or extranodal marginal zone lymphoma. In certain embodiments, the cancer is myeloma, such as multiple myeloma.

[0154] In certain embodiments, the cancer is melanoma, such as advanced melanoma. In certain embodiments, the cancer is progressive or inoperable melanoma that does not respond to other therapies. In certain embodiments, the cancer is melanoma having a BRAF mutation (e.g., BRAF V600 mutation).

[0155] In certain embodiments, the cancer is squamous cell carcinoma of the head and neck (SCCHN). In certain embodiments, the cancer is recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN) including recurrent or metastatic PD-L1+ SCCHN.

[0156] In certain embodiments, the cancer includes a tumor. In certain embodiments, the cancer includes a locally advanced tumor, a metastatic solid tumor, or a combination thereof. In certain embodiments, the cancer includes a tumor for which a PD-1 inhibitor has been shown.

[0157] In certain embodiments, the subject has received prior treatment with at least one anti-PD-1 / anti-PDL-1 therapy. In certain embodiments, the subject has cancer that has failed prior PD-1 / PDL-1 inhibitor therapy. In certain embodiments, the subject has cancer that progresses during prior PD-1 / PDL-1 inhibitor therapy. In certain embodiments, the cancer includes tumors having acquired resistance to anti-PD-1 therapy. In certain embodiments, the cancer includes tumors having innate resistance to anti-PD-1 therapy. In certain embodiments, the cancer includes tumors having resistance to anti-PD-1 therapy and acquired resistance to anti-PD-1 therapy. In certain embodiments, the subject has not received anti-PDL-1 therapy. In certain embodiments, the tumor has a minimum LAG-3 expression of 5% or more of LAG-3 positive immune cells (e.g., lymphocytes and macrophages) compared to all nucleated cells within the tumor region, as shown by an appropriate immunohistochemistry assay. The tumor region includes tumor cells, intratumoral stroma, and peritumoral stroma and does not include normal and / or adjacent uninvolved tissue. Appropriate LAG-3 immunohistochemistry assays are found in Wojcik et al., Consistent Measurement of LAG-3 expression Across Multiple Staining Platforms with the 17B4 Antibody Clone, bioRxiv (February 22, 2022), doi.org / 10.1101 / 2022.02.21.481075, and Johnson et al., Development of a LAG-3 Immunohistochemistry Assay for Melanoma, bioRxiv (February 26, 2022), doi.org / 10.1101 / 2022.02.25.481964.

[0158] In certain embodiments, administration or treatment of the present disclosure produces at least one therapeutic effect. In certain embodiments, the therapeutic effect is a decrease in tumor size. In certain embodiments, the therapeutic effect is a decrease in the number of metastatic lesions over time. In certain embodiments, the therapeutic effect is complete response (e.g., disappearance of all target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is partial response (e.g., a decrease of ≧30% in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is stable disease (e.g., a decrease of ≦30% and an increase of ≦20% in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is progressive disease (e.g., an increase of 20% or more in the sum of the diameters of target lesions according to RECIST v1.1 criteria). In certain embodiments, the therapeutic effect is evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) criteria. In certain embodiments, the therapeutic effect is evaluated according to RECIST v1.1 criteria.

[0159] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used in combination with other therapeutic agents (e.g., anti-cancer agents) that include other molecules (e.g., antibodies) that bind to a target antigen (e.g., a cancer-related antigen). In certain embodiments, the additional therapeutic agent is a chemotherapeutic agent, a radiation therapy agent, or a checkpoint targeting agent. In certain embodiments, the chemotherapeutic agent is a hypomethylating agent (e.g., azacitidine). In certain embodiments, the checkpoint targeting agent is selected from an anti-CTLA-4 antibody, an antagonist anti-PDL-1 antibody, an antagonist anti-PDL-2 antibody, an antagonist anti-PD-1 antibody, an antagonist anti-TIM-3 antibody, an antagonist anti-LAG-3 antibody, an antagonist anti-CEACAM1 antibody, an agonist anti-CD137 antibody, an antagonist anti-CD73 antibody, an antagonist anti-CD96 antibody, an antagonist anti-TIGIT antibody, an antagonist anti-VISTA antibody, an agonist anti-GITR antibody, and an agonist anti-OX40 antibody. In certain embodiments, the additional therapeutic agent is a molecule that binds to 5T4, B7H3, CD19, CD20, CD51, CD123, DR5, EGFR, EpCam, GD2, gpA33, HER2, ROR-1, TAG-72, VEGF-A antibody, and / or VEGFR2.

[0160] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used in combination with one or more immunogenic agents (e.g., tumor vaccines), such as those that target, for example, MUC1, HPV, CEA, and transcription factors (e.g., Twist and lobules). Examples of tumor vaccines include purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), autologous tumor cells, and / or allogeneic tumor cells.

[0161] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used in combination with one or more immunostimulants (e.g., antibodies that activate host immunoreactivity to increase the level of T cell activation). Examples of immunostimulants include anti-PD-1 antibodies, anti-PDL-1 antibodies, and / or anti-CTLA4 antibodies, antibodies against molecules on the surface of dendritic cells (DCs) that activate DC function and antigen presentation, anti-CD40 antibodies that can replace T cell helper activity, activating antibodies against T cell costimulatory molecules (e.g., PDL-1, CTLA-4, OX-40, 4-1BB, ICOS), and / or stimulatory chimeric antigen receptors (CARs).

[0162] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used in combination with one or more compounds that target immunomodulatory enzymes such as IDO (indoleamine-(2,3)-dioxygenase) and / or TDO (tryptophan 2,3-dioxygenase). Examples of compounds that target immunomodulatory enzymes (such as IDO and / or TDO) include epacadostat (Incyte Corp), F001287 (Flexus Biosciences / BMS), indoximod (NewLink Genetics), and NLG919 (NewLink Genetics).

[0163] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are used in combination with one or more antibody-cytokine fusion proteins, also known as immunocytokines (see, e.g., Runbeck et al., Immunocytokines for Cancer, Antibodies 10(10), 2021). Examples of immunocytokines include lymphotoxin, interferon, TNF, TGFβRII, granulocyte macrophage colony-stimulating factor (GM-CSF), IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-12, or NHS-IL12, IL-13, IL-15 or IL-15 superagonist, IL-17, CXCR1 / 2 inhibitor, granulocyte colony-stimulating factor (G-CSF), IL-1β, vascular endothelial growth factor (VEGF), C-C motif chemokine ligand 21 (CCL21), and immunocytokines containing 4-1BBL.

[0164] In certain embodiments, the combination therapies, methods, and kits of the present disclosure are delivered using delivery systems known in the art. Examples of delivery systems include liposomes, microparticles, microcapsules, encapsulation into recombinant cells (e.g., antibodies or antibody fragments) capable of expressing the active pharmaceutical agent, and / or receptor-mediated endocytosis.

[0165] In certain embodiments, the active pharmaceutical agent, combination therapy, or composition of the present disclosure is administered to a subject using one or more methods of administration. Methods of administration of the present disclosure include, but are not limited to, parenteral administration (e.g., intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous), epidural, and mucosal (e.g., intranasal and oral routes). In certain embodiments, the active pharmaceutical agent, combination therapy, or composition is administered intramuscularly, intravenously, or subcutaneously. In certain embodiments, the active pharmaceutical agent, combination therapy, or composition is administered by infusion, bolus injection, or absorption through the epithelium or mucosal lining (e.g., oral mucosa and / or intestinal mucosa). In certain embodiments, the active pharmaceutical agent, combination therapy, or composition is administered together with other bioactive agents. Administration can be systemic or local.

[0166] In certain embodiments, the subject is administered a therapeutically effective amount of an active agent, combination therapy, or composition of the present disclosure. The effective amount can be a single administration or multiple administrations. In some embodiments, the therapeutically effective amount of the active agent, combination therapy, or composition of the present disclosure is administered simultaneously or sequentially.

[0167] In certain embodiments, the anti-PD-1 active agent is administered simultaneously with the anti-TIM-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered simultaneously with the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered simultaneously with the anti-TIM-3 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered prior to the anti-TIM-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered prior to the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered prior to the anti-TIM-3 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered after the anti-TIM-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered after the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered after the anti-TIM-3 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered before the anti-TIM-3 active agent and after the anti-LAG-3 active agent. In certain embodiments, the anti-PD-1 active agent is administered before the anti-LAG-3 active agent and after the anti-TIM-3 active agent.

[0168] In certain embodiments, the anti-TIM-3 active agent is administered concurrently with the anti-PD-1 active agent. In certain embodiments, the anti-TIM-3 active agent is administered concurrently with the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered concurrently with the anti-PD-1 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered prior to the anti-PD-1 active agent. In certain embodiments, the anti-TIM-3 active agent is administered prior to the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered prior to the anti-PD-1 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered subsequent to the anti-PD-1 active agent. In certain embodiments, the anti-TIM-3 active agent is administered subsequent to the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered subsequent to the anti-PD-1 active agent and the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered prior to the anti-PD-1 active agent and subsequent to the anti-LAG-3 active agent. In certain embodiments, the anti-TIM-3 active agent is administered prior to the anti-LAG-3 active agent and subsequent to the anti-PD-1 active agent.

[0169] In certain embodiments, the anti-LAG-3 agent is administered concurrently with the anti-TIM-3 agent. In certain embodiments, the anti-LAG-3 agent is administered concurrently with the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered concurrently with the anti-TIM-3 agent and the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered prior to the anti-TIM-3 agent. In certain embodiments, the anti-LAG-3 agent is administered prior to the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered prior to the anti-TIM-3 agent and the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered after the anti-TIM-3 agent. In certain embodiments, the anti-LAG-3 agent is administered after the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered after the anti-TIM-3 agent and the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered before the anti-TIM-3 agent and after the anti-PD-1 agent. In certain embodiments, the anti-LAG-3 agent is administered before the anti-PD-1 agent and after the anti-TIM-3 agent.

[0170] Anti-PD-1 Administration In certain embodiments, the combination therapies, methods, and kits of the disclosure comprise a therapeutically effective amount of an anti-PD-1 agent.

[0171] In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 1 mg, at least about 5 mg, at least about 10 mg, at least about 20 mg, at least about 50 mg, at least about 100 mg, at least about 150 mg, at least about 200 mg, at least about 250 mg, at least about 300 mg, at least about 350 mg, at least about 400 mg, at least about 450 mg, at least about 500 mg, at least about 550 mg, at least about 600 mg, at least about 650 mg, at least about 700 mg, at least about 750 mg, at least about 800 mg, at least about 850 mg, at least about 900 mg, at least about 950 mg, or at least about 1000 mg of at least one anti-PD-1 agent, and comprises.

[0172] In certain embodiments, an anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 1 to about 5 mg, about 5 to about 10 mg, about 10 to about 20 mg, about 20 to about 50 mg, about 50 to about 100 mg, about 100 to about 150 mg, about 150 to about 200 mg, about 175 to about 225 mg, about 200 to about 250 mg, about 225 to about 275 mg, about 250 to about 300 mg, about 275 to about 325 mg, about 300 to about 350 mg, about 325 to about 375 mg, about 350 to about 400 mg, about 375 to about 425 mg, about 400 to about 450 mg, about 425 to about 475 mg, about 450 to about 500 mg, about 475 to about 525 mg, about 500 to about 550 mg, about 525 to about 575 mg, about 550 to about 600 mg, about 575 to about 625 mg, about 600 to about 650 mg, about 625 to about 675 mg, about 650 to about 700 mg, about 675 to about 725 mg, about 700 to about 750 mg, about 725 to about 775 mg, about 750 to about 800 mg, about 775 to about 825 mg, about 800 to about 850 mg, about 825 to about 875 mg, about 850 to about 900 mg, about 875 to about 925 mg, about 900 to about 950 mg, about 925 to about 975 mg, about 950 to about 1000 mg, about 1000 to about 1050 mg, or about 1000 to about 1100 mg of at least one anti-PD-1 agent. In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 300 to about 600 mg. In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 375 to about 550 mg. In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 450 to about 550 mg. In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 375 to about 500 mg. In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 375 mg.In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 500 mg.

[0173] In certain embodiments, the anti-PD-1 agent (e.g., an anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.2 mg / kg, at least about 0.5 mg / kg, at least about 1 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 5 mg / kg, at least about 10 mg / kg, at least about 20 mg / kg, at least about 30 mg / kg, at least about 50 mg / kg, at least about 75 mg / kg, at least about 100 mg / kg, at least about 125 mg / kg, at least about 150 mg / kg, or greater than 150 mg / kg, based on the weight of the subject.

[0174] In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) as a single dose. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) in two or more doses. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) in one or more doses on a repeated basis. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) three times a day, twice a day, once a day, once every two days, once every three days, once a week, twice a week, three times a week, four times a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once a month, twice a month, three times a month, four times a month, once every two months, or once every three months. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) once every three weeks or once every four weeks. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) once every three weeks. In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) once every four weeks.

[0175] In certain embodiments, the anti-PD-1 agent (e.g., anti-PD-1 antibody or antibody fragment) is administered (or formulated for administration) intravenously.

[0176] Suitable pharmaceutical compositions comprising an anti-PD-1 active agent relate to pharmaceutical compositions comprising an anti-PD-1 active agent, and in certain embodiments disclosed in U.S. Patent Application No. 63 / 220,006, the disclosure of which is incorporated herein by reference (and subsequent patent applications claiming priority to U.S. Provisional Patent Application No. 63 / 220,006 as well), the anti-PD-1 active agent (e.g., an anti-PD-1 antibody or antibody fragment, e.g., retifanlimab) is administered (or formulated for administration) in a pharmaceutical composition comprising acetate, sucrose, polysorbate 80 ("PS80"), and water. In certain embodiments, the anti-PD-1 active agent (e.g., an anti-PD-1 antibody or antibody fragment, e.g., retifanlimab) is administered (or formulated for administration) in a pharmaceutical composition comprising acetate, sucrose, polysorbate 80 (PS80), and water and has a pH of from about 4.0 to about 6.5. In certain embodiments, the concentration of the anti-PD-1 agent in the pharmaceutical composition is from about 10 mg / mL to about 100 mg / mL.

[0177] In certain embodiments, the acetate is present at a concentration of from about 5 mM to about 30 mM. In certain embodiments, the acetate comprises sodium acetate, or glacial acetic acid and sodium acetate.

[0178] In certain embodiments of such pharmaceutical compositions, retifanlimab is present at a concentration of from about 10 mg / mL to about 100 mg / mL, from about 20 mg / mL to about 30 mg / mL, or about 25 mg / mL. In certain embodiments, retifanlimab is present at a concentration of from about 10 mg / mL to about 100 mg / mL. In certain embodiments, retifanlimab is present at a concentration of from about 20 mg / mL to about 30 mg / mL. In certain embodiments, retifanlimab is present at a concentration of about 25 mg / mL.

[0179] In certain embodiments of such pharmaceutical compositions, the acetate comprises glacial acetic acid at a concentration of about 0.05 mg / mL to about 0.35 mg / mL and sodium acetate trihydrate at a concentration of about 0.80 mg / mL to about 2.0 mg / mL. In certain embodiments, the acetate comprises glacial acetic acid at a concentration of about 0.18 mg / mL and sodium acetate trihydrate at a concentration of about 0.95 mg / mL.

[0180] In certain embodiments of such pharmaceutical compositions, sucrose is present at a concentration of about 80 mg / mL to about 100 mg / mL, or about 90 mg / mL.

[0181] In certain embodiments of such pharmaceutical compositions, PS80 is present at a concentration of about 0.08 mg / mL to about 0.15 mg / mL, or about 0.1 mg / mL.

[0182] In certain embodiments of such pharmaceutical compositions, the pharmaceutical composition has a pH of about 4.5 to about 5.7, or about 5.1.

[0183] In other embodiments, for an anti-PD-1 active agent, such as retifanlimab, the pharmaceutical composition a) comprises about 5 mM to about 30 mM acetate, about 50 mg / mL to about 130 mg / mL sucrose, about 0.02 mg / mL to about 0.6 mg / mL PS80, and water, and the composition has a pH of about 4.0 to about 6.5, or b) comprises about 7.5 mM to about 20 mM acetate, about 50 mg / mL to about 130 mg / mL sucrose, about 0.05 mg / mL to about 0.6 mg / mL PS80, and water, and the composition has a pH of about 4.0 to about 6.5, or c) comprises about 9 mM to about 11 mM acetate, about 76 mg / mL to about 104 mg / mL sucrose, about 0.08 mg / mL to about 0.53 mg / mL PS80, and water, and the composition has a pH of about 4.5 to about 5.7, or d) comprises about 9 mM to about 11 mM acetate, about 80 mg / mL to about 100 mg / mL sucrose, about 0.08 mg / mL to about 0.15 mg / mL PS80, and water, and the composition has a pH of about 4.5 to about 5.7.

[0184] In other embodiments, the anti-PD-1 active agent, for example, retifanlimab, the pharmaceutical composition comprises about 25 mg / mL of retifanlimab, about 0.18 mg / mL of glacial acetic acid, about 0.95 mg / mL of sodium acetate trihydrate, about 90 mg / mL of sucrose, about 0.1 mg / mL of PS80, and water, and the composition has a pH of about 4.8 to about 5.4. Anti-TIM-3 Administration

[0185] In certain embodiments, the combination therapies, methods, and kits of the present disclosure comprise a therapeutically effective amount of an anti-TIM-3 active agent.

[0186] In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 1 mg, at least about 5 mg, at least about 10 mg, at least about 20 mg, at least about 50 mg, at least about 100 mg, at least about 150 mg, at least about 200 mg, at least about 250 mg, at least about 300 mg, at least about 350 mg, at least about 400 mg, at least about 450 mg, at least about 500 mg, at least about 550 mg, at least about 600 mg, at least about 650 mg, at least about 700 mg, at least about 750 mg, at least about 800 mg, at least about 850 mg, at least about 900 mg, at least about 950 mg, or at least about 1000 mg of at least one anti-TIM-3 active agent.

[0187] In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least one anti-TIM-3 agent of about 1 to about 5 mg, about 5 to about 10 mg, about 10 to about 20 mg, about 20 to about 50 mg, about 50 to about 100 mg, about 100 to about 150 mg, about 150 to about 200 mg, about 175 to about 225 mg, about 200 to about 250 mg, about 225 to about 275 mg, about 250 to about 300 mg, about 275 to about 325 mg, about 300 to about 350 mg, about 325 to about 375 mg, about 350 to about 400 mg, about 375 to about 425 mg, about 400 to about 450 mg, about 425 to about 475 mg, about 450 to about 500 mg, about 475 to about 525 mg, about 500 to about 550 mg, about 525 to about 575 mg, about 550 to about 600 mg, about 575 to about 625 mg, about 600 to about 650 mg, about 625 to about 675 mg, about 650 to about 700 mg, about 675 to about 725 mg, about 700 to about 750 mg, about 725 to about 775 mg, about 750 to about 800 mg, about 775 to about 825 mg, about 800 to about 850 mg, about 825 to about 875 mg, about 850 to about 900 mg, about 875 to about 925 mg, about 900 to about 950 mg, about 925 to about 975 mg, about 950 to about 1000 mg, about 1000 to about 1050 mg, or about 1000 to about 1100 mg. In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 300 to about 1000 mg. In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 350 to about 500 mg. In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 350 to about 450 mg. In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 400 mg, about 500 mg, or about 1000 mg. In certain embodiments, the anti-TIM-3 agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 400 mg.In certain embodiments, an anti-TIM-3 agonist agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 500 mg. In certain embodiments, an anti-TIM-3 agonist agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 1000 mg.

[0188] In certain embodiments, an anti-TIM-3 agonist agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.2 mg / kg, at least about 0.5 mg / kg, at least about 1 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 5 mg / kg, at least about 10 mg / kg, at least about 20 mg / kg, at least about 30 mg / kg, at least about 50 mg / kg, at least about 75 mg / kg, at least about 100 mg / kg, at least about 125 mg / kg, at least about 150 mg / kg, or greater than 150 mg / kg, based on the body weight of the subject.

[0189] In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) as a single dose. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) in two or more doses. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) in one or more doses on a repeated basis. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) three times a day, twice a day, once a day, once every two days, once every three days, once a week, twice a week, three times a week, four times a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once a month, twice a month, three times a month, four times a month, once every two months, or once every three months. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) once a week, once every two weeks, or once every three weeks. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) once every two weeks or once every three weeks. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) once every two weeks. In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) once every three weeks.

[0190] In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) intravenously.

[0191] In certain embodiments, the anti-TIM-3 active agent (e.g., an anti-TIM-3 antibody or antibody fragment) is administered (or formulated for administration) in a pharmaceutical composition comprising sodium citrate, sucrose, arginine, and polysorbate 80 and has a pH of 6.0. In certain embodiments, the concentration of the anti-TIM-3 agent in the pharmaceutical composition is from about 45 to about 55 mg / mL. In certain embodiments, the concentration of the anti-TIM-3 agent in the pharmaceutical composition is about 50 mg / mL.

[0192] Anti-LAG-3 Administration In certain embodiments, the combination therapies, methods, and kits of the present disclosure comprise a therapeutically effective amount of an anti-LAG-3 active agent.

[0193] In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 1 mg and comprises at least one anti-LAG-3 active agent of at least about 5 mg, at least about 10 mg, at least about 20 mg, at least about 50 mg, at least about 100 mg, at least about 150 mg, at least about 200 mg, at least about 250 mg, at least about 300 mg, at least about 350 mg, at least about 400 mg, at least about 450 mg, at least about 500 mg, at least about 550 mg, at least about 600 mg, at least about 650 mg, at least about 700 mg, at least about 750 mg, at least about 800 mg, at least about 850 mg, at least about 900 mg, at least about 950 mg, or at least about 1000 mg.

[0194] In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least one anti-LAG-3 active agent of about 1 to about 5 mg, about 5 to about 10 mg, about 10 to about 20 mg, about 20 to about 50 mg, about 50 to about 100 mg, about 100 to about 150 mg, about 150 to about 200 mg, about 175 to about 225 mg, about 200 to about 250 mg, about 225 to about 275 mg, about 250 to about 300 mg, about 275 to about 325 mg, about 300 to about 350 mg, about 325 to about 375 mg, about 350 to about 400 mg, about 375 to about 425 mg, about 400 to about 450 mg, about 425 to about 475 mg, about 450 to about 500 mg, about 475 to about 525 mg, about 500 to about 550 mg, about 525 to about 575 mg, about 550 to about 600 mg, about 575 to about 625 mg, about 600 to about 650 mg, about 625 to about 675 mg, about 650 to about 700 mg, about 675 to about 725 mg, about 700 to about 750 mg, about 725 to about 775 mg, about 750 to about 800 mg, about 775 to about 825 mg, about 800 to about 850 mg, about 825 to about 875 mg, about 850 to about 900 mg, about 875 to about 925 mg, about 900 to about 950 mg, about 925 to about 975 mg, about 950 to about 1000 mg, about 1000 to about 1050 mg, or about 1000 to about 1100 mg. In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 300 to about 800 mg. In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 350 to about 750 mg. In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 300 to about 400 mg. In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 350 mg, about 450 mg, or about 750 mg. In certain embodiments, an anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 350 mg.In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 450 mg. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of about 750 mg.

[0195] In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) at a dose of at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.2 mg / kg, at least about 0.5 mg / kg, at least about 1 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 5 mg / kg, at least about 10 mg / kg, at least about 20 mg / kg, at least about 30 mg / kg, at least about 50 mg / kg, at least about 75 mg / kg, at least about 100 mg / kg, at least about 125 mg / kg, at least about 150 mg / kg, or more than 150 mg / kg based on the weight of the subject.

[0196] In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) as a single dose. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) in more than two doses. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) in one or more doses on a repeated basis. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) three times a day, twice a day, once a day, once every two days, once every three days, once a week, twice a week, three times a week, four times a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once a month, twice a month, three times a month, four times a month, once every two months, or once every three months. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) once a week, once every two weeks, or once every three weeks. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) once every two weeks or once every three weeks. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) once every two weeks. In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) once every three weeks.

[0197] In certain embodiments, the anti-LAG-3 active agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) intravenously.

[0198] In certain embodiments, an anti-LAG-3 agent (e.g., an anti-LAG-3 antibody or antibody fragment) is administered (or formulated for administration) in a pharmaceutical composition comprising sodium acetate, trehalose, and polysorbate 80 and has a pH of 5.5. In certain embodiments, the concentration of the anti-LAG-3 agent in the pharmaceutical composition is from about 45 to about 55 mg / mL. In certain embodiments, the concentration of the anti-LAG-3 agent in the pharmaceutical composition is about 50 mg / mL.

[0199] III. Definitions Throughout the present disclosure, substituents or properties of the compounds of the present disclosure are disclosed in groups or ranges. The present disclosure includes each and every individual or partial combination of the members of such groups and ranges, and such groups or ranges are intended to include the endpoints. By way of non-limiting example, if a group or range is from about 1 to about 10, the group or range includes both the value of about 1 and the value of about 10.

[0200] Unless otherwise indicated, the following terms and phrases have the meanings set forth below. The definitions are not intended to be limiting in nature, but rather serve to provide a clearer understanding of certain aspects of the present disclosure.

[0201] Administration: As used herein, the term "administration" refers to providing a composition to a subject.

[0202] Agonist: As used herein, the term "agonist" refers to a ligand, including but not limited to an antigen-binding protein such as an antibody or antibody fragment, that when contacted with a receptor causes one or more of the following: (1) stimulates or activates the receptor, (2) enhances the activity, function, or presence of the receptor, induces or prolongs potentiation or upregulation, (3) mimics one or more functions of a natural ligand or molecule that targets or interacts with the receptor, initiating one or more signal transduction events through the receptor, mimicking one or more functions of the natural ligand, or initiating one or more partial or complete conformational changes seen in known functions or signal transduction through the receptor, and / or (4) enhances, promotes, or induces the expression of the receptor. Agonist activity can be measured in vitro by various assays known in the art, including but not limited to measurement of cell signaling, cell proliferation, immune cell activation markers, or cytokine production. Agonist activity can also be measured in vivo by various assays that measure surrogate endpoints, including but not limited to measurement of T cell proliferation or cytokine production.

[0203] Antagonist: As used herein, the term "antagonist" refers to a ligand, including but not limited to an antigen-binding protein such as an antibody or antibody fragment, that when contacted with a receptor causes at least one of the following: (1) attenuates, blocks, or inactivates the receptor and / or blocks activation of the receptor by its natural ligand, (2) reduces, diminishes, or shortens the activity, function, or presence of the receptor, and / or (3) reduces, diminishes, or abrogates the expression of the receptor. Antagonist activity can be measured in vitro by various assays known in the art, including but not limited to measurement of increases or decreases in cell signaling, cell proliferation, immune cell activation markers, or cytokine production. Antagonist activity can also be measured in vivo by various assays that measure surrogate endpoints, including but not limited to measurement of T cell proliferation or cytokine production.

[0204] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In certain embodiments, "animal" refers to a human at any stage of development. In certain embodiments, "animal" refers to a non-human animal at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, non-human primate, or pig). In certain embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and parasites. In certain embodiments, the animal is a transgenic animal, a genetically engineered animal, or a clone.

[0205] Approximately / About: As used herein, the terms "approximately" and "about" are used interchangeably herein and refer to a value within + / - 10% of the recited value applied to one or more values of interest. In certain embodiments, this term refers to a range of values within + / - 10%, + / - 9%, + / - 8%, + / - 7%, + / - 6%, + / - 5%, + / - 4%, + / - 3%, + / - 2%, + / - 1%, or less of the recited reference value, unless otherwise explicitly stated or otherwise apparent from the context.

[0206] Binding affinity: As used herein, the term "binding affinity" describes the total strength of non-covalent interactions between a binding site of a molecule (e.g., an antibody) and a binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (K D )(e.g., "lower affinity" referring to a larger K D ). Affinity can be measured and / or expressed in many ways known in the art, (i) the equilibrium dissociation constant (K D ) calculated from the quotient of k-off / k-on, and (ii) the equilibrium association constant (K A) include, but are not limited to. "k-on" refers to, for example, the association rate constant of an antibody to an antigen, and "k-off" refers to, for example, the dissociation rate constant of an antibody from an antigen. k-on and k-off can be determined by techniques known to those skilled in the art, such as Biacore assays or KinExA.

[0207] Complete response: As used herein, the term "complete response" or "CR" refers to the removal of all or substantially all target lesions at the target. In certain embodiments, CR refers to a reduction of about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% in the total diameter of the target lesions (i.e., loss of lesions) compared to the baseline total diameter. In certain embodiments, CR refers to about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the total lesion diameter remaining after treatment compared to the baseline total lesion diameter. In certain embodiments, complete response is evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines.

[0208] Epitope: As used herein, the term "epitope" refers to a local region of an antigen to which an antibody (or a fragment thereof) can bind. An epitope can be, for example, a continuous amino acid of a polypeptide (i.e., a linear or continuous epitope), or can also include a plurality of non-continuous amino acids of a polypeptide chain that together form an epitope region from at least two non-continuous regions of the polypeptide.

[0209] Epitope-binding fragment: As used herein, the term "epitope-binding fragment" describes a portion or fragment of an antibody that can bind to an epitope.

[0210] Epitope-binding site: As used herein, the term "epitope-binding site" refers to a part of a molecule that includes an epitope-binding fragment involved in binding to a target epitope.

[0211] Function: As used herein, "function" refers to a characteristic, property, or distinctive element of an identified object.

[0212] Identity: As used herein, the terms "identity" and "percent identity" refer to the overall relatedness between the sequences of polymeric molecules, e.g., polynucleotide molecules or polypeptide molecules. The calculation of the percent identity between two sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced into one or both of the sequences for optimal alignment), and then comparing the aligned sequences. The percent identity between two sequences is a function of the number of identical positions shared by the sequences relative to the total length of the sequences, taking into account the number and length of any gaps introduced for optimal alignment of the sequences. In certain embodiments, the comparison of sequences and determination of the percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two sequences can be determined using the methods described in Karlin et al., PNAS (1990) 87: 2264-2268, Karlin et al., PNAS (1993) 90: 5873-5877, Carillo et al., Applied Math. (1988) 48:1073, Meyers et al., CABIOS (1989) 4:11-17 (incorporated into the ALIGN program), Lesk et al., Computational Molecular Biology, Oxford University Press, New York, 1988, Smith et al., Biocomputing: Informatics and Genome Projects, Academic Press, New York, 1993; Von Heinje et al., Sequence Analysis in Molecular Biology, Academic Press, 1987; Griffin et al., Computer Analysis of Sequence Data, Part I, Humana Press, New Jelon, 1994; and Gribskov et al., Sequence Analysis Primer, Stockton Press, New York, 1991.As an example, the percent identity between two sequences can be determined using the GAP program in the GCG software package using the NWSgapdna.CMP matrix. Techniques and tools for determining percent identity are also standardized by publicly available computer programs including, but not limited to, the GCG program package, FASTA, P-BLAST, N-BLAST, and X-BLAST (e.g., National Center for Biotechnology Information (NCBI) on the world wide web, ncbi.nlm.nih.gov).

[0213] Partial response: As used herein, the term "partial response" or "PR" refers to a decrease in tumor progression in a subject, as indicated by a decrease in the sum of the diameters of the target lesions relative to the baseline sum diameter. In certain embodiments, PR refers to at least a 30% decrease in the sum of the diameters of the target lesions as compared to the baseline sum diameter. In certain embodiments, PR refers to at least about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or at least about 90% decrease in the sum of the diameters of the target lesions relative to the baseline sum diameter. In certain embodiments, partial response is evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines.

[0214] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" or "therapeutically acceptable" is used to describe a compound, material, composition, and / or dosage form that is suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0215] Pharmaceutically acceptable excipient: As used herein, the term "pharmaceutically acceptable excipient" or "therapeutically acceptable excipient" refers to a component in a composition other than the active agent described herein (e.g., a vehicle capable of suspending, supporting, or encapsulating the active agent), and has the property of being substantially non-toxic and non-inflammatory in a subject.

[0216] Progression: As used herein, the term "progressive disease" or "PD" refers to an increase in tumor progression in a subject, as indicated by an increase in the total diameter of target lesions relative to the total baseline diameter. In certain embodiments, PD refers to an increase of at least 20% in the total diameter of target lesions relative to the total baseline diameter. In certain embodiments, PR refers to an increase of at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or at least about 90% in the total diameter of target lesions compared to the baseline total diameter. In certain embodiments, PD refers to the appearance of one or more new lesions. In certain embodiments, progressive disease is evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines.

[0217] Stable: As used herein, the term "stable disease" or "SD" refers to tumor progression that has neither a reduction sufficient to be considered a partial response (PR) nor an increase sufficient to be considered progressive disease (PD). In certain embodiments, SD refers to a decrease of less than 30% in the total diameter of target lesions compared to the total baseline diameter. In certain embodiments, SD refers to a decrease of less than about 30%, 25%, 20%, 15%, 10%, or 5% in the sum of the diameters of target lesions relative to the baseline sum of diameters, or a 0% decrease. In certain embodiments, SD refers to an increase of less than 20% in the total diameter of target lesions compared to the total baseline diameter. In certain embodiments, SD refers to an increase of less than about 20%, 15%, 10%, or 5% in the sum of the diameters of target lesions relative to the baseline sum of diameters, or a 0% increase. In certain embodiments, stable disease is evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines.

[0218] Subject: As used herein, the term "subject" refers to any organism to which a composition according to the present disclosure can be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (such as mammals like mice, rats, rabbits, non-human primates, and humans) and / or plants. A subject or patient may be seeking treatment, in need of treatment, requiring treatment, receiving treatment, or undergoing care by a trained professional for a particular disease or condition.

[0219] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" of a drug is an amount sufficient to produce a beneficial or desired result (such as a biological, medical, or clinical result). Thus, the effective amount depends on the circumstances to which it is applied (such as the route of administration, the severity of the condition, biochemistry, and the medical history of the subject, etc.) and can be determined by standard clinical techniques by those skilled in the art (such as extrapolated from a dose-response curve derived from testing). For example, in the context of administering a drug to treat cancer, the effective amount of the drug can be an amount sufficient to achieve treatment of the cancer as compared to the response obtained without administration of the drug.

[0220] Treatment: As used herein, the term "treatment" refers to, for example, the partial or complete alleviation, amelioration, improvement, remission, prevention, delay in onset, inhibition of progression, reduction in severity, and / or reduction in incidence of one or more symptoms or features of a particular disease, disorder, and / or condition, such as cancer. Treatment may be administered to a subject that does not exhibit signs of a disease, disorder, and / or condition and / or to a subject that exhibits early signs of a disease, disorder, and / or condition for the purpose of reducing the risk of developing the disease, disorder, and / or condition-related symptoms or pathology, for example, reducing tumor size, reducing the presence of circulating tumor cells, reducing or preventing metastasis, slowing or halting tumor growth, and / or preventing or delaying tumor recurrence or relapse. Treatment may also include improvement of one or more features related to a disease, disorder, and / or condition beyond a baseline, including improvement relative to measurements or observations made prior to initiation of treatment by the methods of the present disclosure.

[0221] General Considerations One of ordinary skill in the art will recognize, or be able to ascertain using no more than routine experimentation, one or more equivalents to the specific embodiments of the disclosure described herein. The scope of the disclosure is not intended to be limited to the above description.

[0222] The articles "a", "an", and "the" can mean one or more than one, unless indicated to the contrary or not apparent from the context. A claim or description that includes "or" among one or more members of a group means that one, two, or all of the group members are present in, employed in, or otherwise related to a given product or process and are considered satisfied, unless indicated to the contrary or not apparent from the context. This disclosure can include embodiments where exactly one member of a group is present in, used in, or otherwise related to a given product or process. This disclosure can also include embodiments where two or more, or all members of a group are present in, used in, or otherwise related to a given product or process.

[0223] The term "comprising" is intended to be open-ended and permissive and does not require the inclusion of additional elements or steps. When the term "comprising" is used herein, the terms "consisting of" and "consisting essentially of" are also included and disclosed.

[0224] Abbreviations, e.g., are derived from exemplary Latin phrases and are used herein to provide non-limiting examples. Thus, the abbreviation "e.g." is synonymous with the term "for example". The abbreviation "i.e." is derived from the Latin id est and is used herein to provide non-limiting rephrasing or clarification. Thus, the abbreviation "i.e." is synonymous with the term "that is".

[0225] Any particular embodiment of this disclosure that is included in the prior art can be explicitly excluded from any one or more of the claims. Any particular embodiment of the agents, combination therapies, methods, and / or compositions of this disclosure can be excluded from any one or more of the claims for any reason, whether or not related to the existence of the prior art.

[0226] This specification incorporates by reference publications, patent applications, patents, and other references mentioned herein, and prevails in case of conflict with this specification.

[0227] The headings of sections, materials, methods, and examples are illustrative only and are not intended to be limiting.

Examples

[0228] Example 1 Combination Therapy Trial of Anti-PD-1, Anti-TIM-3, and Anti-LAG-3 Active Agents in Urothelial Cancer Trials have been conducted to investigate the biological and therapeutic outcomes of selected combination therapies that include combinations of anti-PD-1, anti-TIM-3, and anti-LAG-3 agents. This trial is also being conducted to evaluate the clinical and biological effects of blocking LAG-3 and TIM-3, additional important checkpoint pathways involved in the lack of / loss of response to PD-1 inhibitors.

[0229] A cohort of 18 patients was selected who had muscle-invasive urothelial carcinoma of the bladder who had undergone (or had previously undergone) radical cystectomy and who were either cisplatin-ineligible or had refused cisplatin therapy.

[0230] The anti-PD-1 antibody in this trial is INCMGA00012 (i.e., retifanlimab). The anti-TIM-3 antibody for the trial is Antibody A. The anti-LAG-3 antibody for the trial is Antibody B. The dosing sequence of the investigational agents is (1) INCMGA00012, (2) Antibody B, followed by (3) Antibody A.

[0231] a. Selection of the Dose of INCMGA00012 On Day 1 of INCMGA00012, their drug (i.e., retifanlimab) is administered by IV infusion over approximately 30 minutes at 500 mg once every 4 weeks (i.e., Q4W). The dosing formulation contains 25 mg / mL in a 500 mg dose liquid formulation.

[0232] This dose was selected based on modeling of clinical PK data from the first monotherapy trial in humans where 37 patients were administered doses of 1 mg / kg Q2W, 3 mg / kg Q2W, 3 mg / kg Q4W, 10 mg / kg Q2W, and 10 mg / kg Q4W. Pharmacokinetic data were obtained from 15 patients who received INCMGA00012 500 mg Q4W. The AUC 0-∞ observed at 500 mg Q4W approximates the steady-state AUC 0-t based on population PK analysis of body weight-adjusted doses, and the estimated clearance is similar. The probability that plasma trough concentration at steady state will be 21 μg / mL or higher (resulting in maximum target engagement and highest probability of efficacy) with administration of 500 mg Q4W is approximately 58%. Based on the above observations, 500 mg Q4W was selected as the dosing regimen.

[0233] b. Selection of the Dose of Antibody B Antibody B is administered by IV infusion over approximately 30 minutes on Day 1 of each 14-day cycle at 350 mg, once every two weeks (i.e., Q2W). The dosing formulation contains 50 mg / mL in a liquid formulation of 350 mg dose.

[0234] This dose selection is based on modeling of clinical PK data from human monotherapy trials, where 22 participants were treated in an open-label, non-randomized, dose-escalation, and cohort expansion trial with IV doses of 25, 75, 250, 350, and 750 mg Q2W. Tolerability was good at all doses. Antibody B PK was linear after the first dose and showed moderate to high inter-individual variability. At a dose of 350 mg Q2W (N = 3), the steady-state AUC was 30,200 μg / mL·h. The 350 mg Q2W dose was selected based on data indicating that (i) the receptor on the cell surface was fully occupied at trough concentration after 350 mg Q2W administration and (ii) the dose was pharmacologically active as measured by increased peripheral T cell activation observed at doses above 250 mg.

[0235] c. Selection of the Dose of Antibody A Antibody A is administered by IV infusion over approximately 30 minutes on Day 1 of each 14-day cycle, once every two weeks (i.e., Q2W) at a dose of 400 mg. The Q2W dosing formulation contains 50 mg / mL in a liquid formulation for the 400 mg dose.

[0236] This dose selection is based on modeling of clinical PK data from a human monotherapy trial in which 38 participants were treated with IV doses of 10, 30, 100, 200, 400, 800, and 1600 mg Q2W in an open-label, non-randomized, dose-escalation, and cohort expansion trial. Tolerability was good at all doses. Antibody A demonstrated supra-proportional PK after the first dose of 30 mg to 800 mg Q2W, and the inter-individual variability in PK exposure was low. At the 400 mg Q2W dose, the mean steady-state AUC determined in 4 participants was 43,400 μg / mL·h. The 400 mg dose was selected based on data showing that the TIM-3 receptor was fully occupied on the surface of circulating monocytes at trough concentrations after 400 mg Q2W dosing. Furthermore, changes in pharmacodynamic markers suggest that this dose is pharmacologically active.

[0237] d. Selection of Patients Tumor tissue from each patient was tested to determine the PD-L1 CPS score. Participants were randomized to treatment groups, with participants having a PD-L1 CPS of less than 10 or a PD-L1 CPS of 10 or greater being equally randomized to the treatment groups.

[0238] The inclusion criteria for patients were as follows: (i) male or female aged 18 years or older, (ii) participants with histologically confirmed urothelial carcinoma of the transitional cell - having mixed histology had to have a predominant (i.e., at least 50%) transitional cell pattern, (iii) clinical stage T2 - T3b, N0, M0 muscle - invasive urothelial carcinoma by CT (or MRI) (stages II - IIIA according to AJCC 2018) - participants with concurrent upper tract tumors were excluded. A previous surgical history of upper tract tumors was allowed, but non - invasive pT (i.e., pT < 2N0M0 stage), (iv) ineligibility for cisplatin therapy, ECOG PS 2 participants were excluded, according to the modified Galsky criteria (see Appendix D), (v) refusal of cisplatin - based therapy, (vi) eligible for radical cystectomy, (vii) 0 or 1 ECOG PS, (viii) pretreatment tumor biopsy had to be a tumor block from a biopsy of the primary tumor containing at least 20% of the tumor or 20 unstained slides.

[0239] Exclusion criteria for patients include the following: (i) Participation in another trial that received investigational drug or device within 28 days or within 5 half-lives (whichever is longer) before the first dose of this trial; (ii) Prior receipt of systemic therapy for bladder cancer or prior treatment with checkpoint inhibitors (such as anti-PD-1, anti-PDL-1, anti-PDL-2, or anti-CTLA4); (iii) Evidence of measurable nodular or metastatic disease; (iv) Concurrent anti-cancer therapy (e.g., chemotherapy, radiotherapy, surgery, immunotherapy, biologic therapy, hormonal therapy, investigational drug, intravesical therapy, or tumor embolization); (v) Undergoing major surgery within 4 weeks prior to registration (C1D1); (vi) Having a known additional malignancy other than miUBC that is progressive or requires active treatment, or having a history of another malignancy within 2 years from trial registration, except for healed basal cell or squamous cell carcinoma of the skin, superficial bladder cancer, prostatic intraepithelial neoplasia, cervical intraepithelial neoplasia, or non-invasive or indolent malignancies, or cancers for which the participant was disease-free for less than 1 year after treatment with intent to cure; (vii) Having an active autoimmune disease that requires systemic immunosuppression with corticosteroids (prednisone or equivalent daily dose > 10 mg) or immunosuppressive agents within 2 years of day 1 of the trial treatment; (viii) Patients with an active autoimmune disease that requires systemic immunosuppression with corticosteroids (> 10 mg / day of prednisone or equivalent) beyond physiological maintenance dose; (ix) Having known active hepatitis B or C or HIV, co-infected with HBV, HCV, or hepatitis D virus, and known to have cancerous meningitis; (x) Having an active infection that requires systemic antibiotics within 14 days from the first dose of the investigational drug; (xi) Known or suspected COVID-19 infection; (xii) Use of probiotics within 28 days from the first dose of the investigational drug; (xiii) Prohibited drugs (e.g., other anti-cancer therapies, immunosuppressive drugs or corticosteroids, systemic steroid drugs, live vaccines, UGT1A9 inhibitors, warfarin, and systemic antibiotics); (xiv) Not having recovered to grade 1 or less from the toxic effects of previous therapy and / or complications from previous surgical intervention before starting the trial therapy.(xv) History of or presence of abnormalities, clinically significant ECG, (xvi) Gastrointestinal conditions that may affect the absorption of oral medications (e.g., inflammatory bowel disease, Crohn's disease, ulcerative colitis), (xvii) Administration of live vaccines within 30 days prior to the planned start of the test therapy, (xvii) Cardiac dysfunction or clinically significant heart disease, (xviii) Previous allogeneic tissue / solid organ transplantation, (xix) Participants who failed the clinical examination screening and had (a) platelets < 100×10, 9 / L, (b) hemoglobin < 9 g / dL, (c) ANC < 1.5×109 / L, (d) ALT > 2×ULN, (e) AST > 2×ULN, (f) CrCl < 30 mL / min, calculated by the Cockcroft - Gault equation, (g) INR or PT > 1.5×ULN (except for therapeutic anticoagulants), (h) aPTT > 1.5×ULN, (xx) Evidence of interstitial lung disease or active non - infectious pneumonia, (xxi) Any test drug, or excipients containing mannitol or another monoclonal antibody that cannot be controlled by standard means (e.g., anti - histamines and corticosteroids), and a known allergy to them, and (xxii) Any patient with immune - related toxicity grade 2 or above and who has previously received immunotherapy were included.

[0240] e. Objectives and Evaluation Items of the Trial The primary study endpoint for each treatment group is the change from baseline in CD8+ lymphocytes within the resected tumor.

[0241] Secondary evaluation items include (i) Safety and tolerability evaluated by monitoring the frequency and severity of adverse events (AEs) including delay of cystectomy due to AEs, (ii) The pCR rate of patients defined as the proportion of participants with ypT0N0 in each treatment group, and (iii) The major pathologic response defined as residual ypT0 / 1 / a / isN0M0.

[0242] The exploratory evaluation items include the following: (i) changes in tumor, tumor stroma, and tumor-associated immune cell gene expression profiles from baseline for each visit, where the variables are measured and associated with treatment outcomes; (ii) changes in tumor, tumor stroma, and tumor-associated immune cell protein and metabolite marker levels and spatial distributions from baseline for each visit, where the variables are measured and associated with treatment outcomes; (iii) changes in plasma analytes including cytokines and other known markers of inflammation and immune status, tumor markers, and markers related to metabolic and nutritional status from baseline for each visit, where the variables are measured and associated with treatment outcomes; (iv) the biological response rate of patients with a CPS of less than 10 versus 10 or more in each treatment group; (v) a descriptive analysis between the combination / consecutive treatment group and the monotherapy treatment group; (vi) pathologic downstaging (i.e., ypT<2N0M0) in each treatment group; (vii) the rate of pathologic downstaging in each treatment group is calculated and its 80% CI is estimated using the Clopper Pearson method (Voskuilen et al., 2019).

[0243] The efficacy analysis of the pCR rate and pathologic downstaging assessment includes all participants who meet the eligibility criteria, receive at least one cycle of investigational treatment, undergo cystectomy, or discontinue due to disease progression. pCR is defined as pT0 and carcinoma in situ based on histological evaluation of TURBT and cystectomy specimens by local institutional analysis. The major pathologic response is defined as residual ypT0 / 1 / a / isN0M0 based on histological evaluation of TURBT and cystectomy specimens. Pathologic downstaging is defined as ypT < 2N0M0 and is performed based on evaluation of TURBT and cystectomy specimens by local institutional analysis using the AJCC staging system 8th edition, 2017.

[0244] Example 2 Combination Therapy Trial of Anti-PD-1, Anti-TIM-3, and Anti-LAG-3 Active Agents in Melanoma Trials are being conducted to investigate the biological and therapeutic outcomes of selected combination therapies that include combinations of anti-PD-1 agents, anti-TIM-3 agents, and anti-LAG-3 agents. This trial is also conducted to evaluate the efficacy of combinations of anti-PD-1 agents, anti-TIM-3 agents, and anti-LAG-3 agents as the latest line of therapy in participants with melanoma that has progressed or recurred on or after anti-PDL-1 therapy.

[0245] Fifteen to thirty patients with progressive malignancies (e.g., unresectable, metastatic, or adjuvant-treated melanoma) are selected as a cohort, who have previously received treatment targeting PD-1 and / or PDL-1 and have deteriorated or recurred after the previous anti-PDL-1 therapy.

[0246] This trial has two phases. The first phase has the following four parts:

[0247] (1a) In the first phase, using the PART 1 - BOIN design, in patients with tumors to which a PD-1 inhibitor is applicable and who have failed PD-1-directed therapy, determine whether the doses selected for the trial are safe when used in combination with antibody B (350 mg Q2W) and antibody A (400 mg Q2W) (n ≤ 9). If the safety of the tested doses is not established, the dose levels are reduced (i.e., for antibody B, 250 mg Q2W; for antibody A, 200 mg Q2W).

[0248] (1b) After confirming the safety of the combination of antibody B and antibody A in the first phase, part 2, in patients with tumors to which a PD-1 inhibitor is applicable and who have failed PD-1-directed therapy, test the safety of the combination of antibody B (350 mg Q2W) + antibody A (400 mg Q2W) + INCMGA00012 (500 mg Q4W) (n ≤ 9). If the safety of the tested doses is not established, the dose levels of antibody B and antibody A are reduced (i.e., for antibody B, 250 mg Q2W; for antibody A, 200 mg Q2W).

[0249] (1c) Safety trial on the following combination therapies in patients with tumors to which the first-phase, part 3 - PD-1 inhibitor is applicable and who have failed PD-1-directed therapy: (1c.i) Antibody B (450 mg Q3W) + INCMGA00012 (375 mg Q3W) (n~6), patients first receive a 28-day cycle of 450 mg of Antibody B and then receive the doublet combination for subsequent cycles, (1c.ii) Antibody B (750 mg Q3W) + INCMGA00012 (375 mg Q3W) (n~6), patients first receive a 28-day cycle of 750 mg of Antibody B and then receive the doublet combination for subsequent cycles, (1c.iii) Antibody A (500 mg Q3W) + INCMGA00012 (375 mg Q3W) (n~6), patients are first administered 500 mg of Antibody A in a 28-day cycle and then receive the doublet combination for subsequent cycles, (1c.iv) Antibody A (1000 mg Q3W) + INCMGA00012 (375 mg Q3W) (n~6), patients first receive a 28-day cycle of 1000 mg of Antibody A and then receive the doublet combination for subsequent cycles.

[0250] (1d) Safety trial on the following combination therapy in patients with tumors to which the first-phase part 4 - PD-1 inhibitor is applicable and who have failed PD-1-directed therapy: Antibody B (Q4W) + Antibody A (Q4W) + 500 mg (Q4W) INCMGA00012. The Q4W dosage of Antibody B and Antibody B is based on pharmacokinetic (PK) and receptor occupancy (RO) data from the Q3W monotherapy of Antibody B and Antibody B in the above first-phase part 3.

[0251] (2) The second phase determines the preliminary efficacy of the combination of Antibody B + Antibody A + INCMGA00012 and is completed in two cohorts.

[0252] (2a) A population of patients (n = 15) with unresectable / metastatic or adjuvant-treated melanoma who progressed or relapsed before and after prior anti-PDL-1 therapy in Cohort A - Phase 2 is given a combination of 350 mg (Q2W) of Antibody B + 400 mg (Q2W) of Antibody A + 500 mg (Q4W) of INCMGA00012. If a sufficient number of positive responses are seen (≥1 / 15 responses), the population is expanded to include additional participants (+n = 15) in Simon's two-stage expansion to further evaluate safety and efficacy.

[0253] (2b) A population of patients with untreated advanced melanoma (1L) in Cohort B - adjuvant (N~34) who may have failed CPI is randomized and administered one of the following combination therapies: (2b.i) (N~17) 450 mg (Q3W) Antibody B + 500 mg (Q3W) Antibody A + 375 mg (Q3W) INCMGA00012, or (2b.ii) (N~17) 750 mg (Q3W) Antibody B + 1000 mg (Q3W) Antibody A + 375 mg (Q3W) INCMGA00012. If a sufficient number of positive responses are seen, the population is expanded to include additional participants (+n = 18 for each cohort) in the expansion to further evaluate safety and efficacy.

[0254] The anti-PD-1 antibody in this trial is INCMGA00012 (i.e., relatlimab). The anti-TIM-3 antibody for the trial is Antibody A. The anti-LAG-3 antibody for the trial is Antibody B. The administration order of the investigational agents is (1) Antibody B, followed by (2) Antibody A, followed by (3) INCMGA00012.

[0255] a. Selection of the Dose of INCMGA00012 INCMGA00012 (i.e., relatlimab) is administered at 500 mg once every 4 weeks (i.e., Q4W) by IV infusion over approximately 30 minutes on Day 1 of each 28-day cycle. The dosage form for administration contains 25 mg / mL in a liquid formulation of 500 mg dose.

[0256] This dose was selected based on modeling of clinical PK data from the first monotherapy trial in humans where 37 patients were administered each of the doses of 1 mg / kg Q2W, 3 mg / kg Q2W, 3 mg / kg Q4W, 10 mg / kg Q2W, and 10 mg / kg Q4W. Pharmacokinetic data were obtained from 15 patients who received INCMGA00012 500 mg Q4W. The AUC 0-∞ observed at 500 mg Q4W approximates the steady-state AUC 0-t based on population PK analysis of body weight-adjusted doses, and the estimated clearance is similar. The probability that plasma trough concentrations at steady state will be 21 μg / mL or greater (resulting in maximum target binding and the highest probability of demonstrating efficacy) with a 500 mg Q4W dose is approximately 58%. Based on these observations, 500 mg Q4W was selected as the dosing regimen.

[0257] Furthermore, 375 mg, once every three weeks (i.e., Q3W), by IV infusion, INCMGA00012 those trials are being conducted. While not wishing to be bound by any theory, this dose was selected because a 375 mg Q3W dose is predicted to achieve the same steady-state trough concentration as a 500 mg Q4W dose.

[0258] b. Selection of the Dose of Antibody B Antibody B is administered at 350 mg, once every two weeks (i.e., Q2W), by IV infusion over approximately 30 minutes on day 1 of each 14-day cycle. The dosing formulation contains 50 mg / mL in a liquid formulation at the 350 mg dose. Antibody B is in a liquid form of sodium acetate, trehalose, polysorbate 80, pH 5.5 and has a target protein concentration of 50 mg / mL for use in IV infusion.

[0259] This dose selection is based on modeling of clinical PK data from human monotherapy trials. Twenty-two participants were treated in an open-label, non-randomized, dose-escalating, and cohort expansion trial with IV doses of 25, 75, 250, 350, and 750 mg Q2W. Tolerability was good at all doses. Antibody B PK was linear after the first dose and showed moderate to high inter-individual variability. At the 350 mg Q2W (N = 3) dose, the steady-state AUC was 30,200 μg / mL·h. The 350 mg Q2W dose was selected based on data indicating that (i) receptors on the cell surface were fully occupied at trough concentrations after 350 mg Q2W dosing and (ii) the dose was pharmacologically active as measured by an increase in peripheral T cell activation observed at doses above 250 mg.

[0260] If the safety of the doses tested in combination therapy trials is not established, the dose level of Antibody B will be decreased by one level from the doses tested in monotherapy trials to 250 mg Q2W.

[0261] Furthermore, Antibody B is also being studied by IV infusion at 450 mg and 750 mg once every three weeks (i.e., Q3W).

[0262] c. Selection of the Dose of Antibody A Antibody A is administered by IV infusion at 400 mg once every two weeks (i.e., Q2W) over approximately 30 minutes on day 1 of each 14-day cycle. The dosage formulation contains 50 mg / mL in a liquid formulation for the 400 mg dose. Antibody A is in a liquid form of sodium citrate, sucrose, arginine, polysorbate 80, pH 6.0 at a target protein concentration of 50 mg / mL for use in IV infusion.

[0263] This dose selection is based on modeling of clinical PK data from human monotherapy trials. Forty participants were treated with Q2W IV doses of 10, 30, 100, 200, 400, 800, and 1600 mg in an open-label, non-randomized, dose-escalating, and cohort-expansion trial. Tolerability was good at all doses. Antibody A showed supra-proportional PK after the first dose of 30 mg to 800 mg Q2W, and the inter-individual variability in PK exposure was low. At a dose of 400 mg Q2W, the mean steady-state AUC determined in four participants was 43,400 μg / mL·h. The 400 mg dose was selected based on data indicating that the TIM-3 receptor was fully occupied at trough concentrations on circulating monocytes after Q2W administration of 400 mg.

[0264] If the safety of the doses tested in combination therapy trials is not established, the dose level of Antibody A will be decreased by one level from the doses tested in monotherapy trials to 200 mg Q2W.

[0265] Furthermore, Antibody A is being studied by IV infusion at 500 mg and 1000 mg once every three weeks (i.e., Q3W).

[0266] d. Selection of Patients Inclusion criteria for patients are as follows: (i) being male or female aged 18 years or older; (ii) participants with locally advanced or metastatic solid tumors for which first-phase PD-1 inhibitors are indicated and who have failed PD-1 / PD-L1 inhibitor therapy as defined below (locally advanced disease should not be able to be resected with curative intent): The subject should have received previous treatment with anti-PD-1 or anti-PD-L1 therapy (alone or as part of combination therapy) in a progressive or metastatic setting, and (ii.a) have PD as the best response to treatment confirmed at least 4 weeks (28 days or more) later, (ii.b) have received at least 2 administrations of anti-PD-1 or anti-PD-L1 agents, (ii.c) progressive disease should also be at least 12 weeks from the first dose of anti-PD-1 or anti-PD-L1 therapy and confirmed 4 weeks (28 days or more) later, and (ii.d) PD should be based on imaging performed at least 4 weeks apart; (iii) second-phase - (iii.a) participants with histologically confirmed unresectable / metastatic melanoma, the disease having previously failed anti-PD-L1 therapy (alone or as part of combination therapy), and the following criteria: (iii.a1) participants who have failed previous adjuvant anti-PD-L1 therapy for resectable melanoma should have received previous anti-PD-L1 for 6 weeks or more and experienced disease progression while receiving active adjuvant therapy containing anti-PD-L1, or participants in whom early recurrence occurred within 24 weeks after the end of adjuvant anti-PD-L1 therapy [progressive disease should be confirmed by a confirmatory biopsy taken at baseline], or (iii.a2) participants in whom unresectable / metastatic disease has progressed within 24 weeks after completion of anti-PD-L1 as the latest therapy for unresectable / metastatic melanoma [progressive disease should be confirmed by imaging ≧4 weeks after the first evidence of disease progression], (iii.b) Participants must have received no more than two prior treatments for melanoma, and the most recent prior treatment must have contained no anti-PD-L1 therapy (either alone or as part of a combination therapy) in either the adjuvant and / or metastatic / progressive setting [Subjects with tumors that are BRAF variants may have received up to two prior treatments for melanoma in addition to previous BRAF-directed therapy] [Participants may have received anti-PD-L1 therapy in both the adjuvant and metastatic / progressive settings], (iii.c) For participants with BRAF-mutant tumors, treatment with a targeted therapy is required, and for eligibility, the participant must be progressing or intolerant to the targeted therapy, (iii.d) The participant must have a fresh biopsy available after completion of the previous PD-(L)1 therapy or be willing and able to undergo a pre-treatment tumor biopsy (core or excisional), (iii.e) The participant must have measurable disease as defined by RECIST v1.1 [Lesions used for response assessment must not be the same lesions used for biopsy, (iii.e1) must not be in a field that has been previously irradiated, or (iii.e2) must meet one of the following: show clear evidence of radiographic progression after completion of previous radiotherapy or biopsy and before study enrollment], (iv) ECOG performance status is 0 or 1, are included.

[0267] Exclusion criteria for patients include the following: (i) Participants who failed clinical screening, (i.a) Platelets - <100×10 9 / L, (i.b) hemoglobin < 9 g / dL, (i.c) ANC < 1.5 × 10⁹ / L, (i.d) ALT > 1.5 × ULN (Phase 1) or > 2.5 × ULN (Phase 2) (i.e., AST > 1.5 × ULN (Phase 1) or > 2.5 × ULN (Phase 2)), (i.f) total bilirubin ≥ 1.5 × ULN, conjugated bilirubin ≤ ULN (conjugated bilirubin needs to be tested only if total bilirubin exceeds ULN). In the absence of a facility ULN, direct bilirubin must be < 40% of total bilirubin, and in any case, total bilirubin cannot exceed 3 × ULN, (i.g) lactate dehydrogenase > 3 × ULN in the absence of hemolysis, (i.h) calculated creatinine clearance < 50 mL / min [CrCl calculated by the Cockcroft - Gault equation], (i.i) international normalized ratio or prothrombin time > 1.5 × ULN, and (i.j) activated partial thromboplastin time > 1.5 × ULN; (ii) having a known allergy or severe reaction to any component of the test agent or even to the formulation components of INCMGA00012 or antibody B or antibody A; (iii) Phase 1 - having received an anticancer agent or test drug within the following intervals prior to the first administration of the test treatment: (iii.a) within 14 days for chemotherapy, targeted small - molecule therapy, or radiotherapy [participants do not require long - term use of corticosteroids and must not have radiation pneumonia as a result of treatment. With the approval of the medical monitor, it is acceptable to wash out palliative radiotherapy for non - CNS diseases for 1 week. Bisphosphonates and denosumab are allowed as concomitant medications], (iii.b) within 14 days and the disappearance of all toxicities related to the persistence of previous immunotherapy or active cell therapy (i.e., chimeric antigen receptor T - cell therapy), (iii.c) less than 14 days for previous PD - 1 pathway - targeted agents, (iii.d) within 28 days for previous mAbs used in anticancer therapy, excluding PD - 1 pathway - targeted agents and denosumab, (iii.e) For immunosuppression-based treatment for any reason within 7 days [use of inhaled or topical steroids or corticosteroids for radiation treatment is permitted], use of physiological corticosteroid replacement therapy may be approved after consultation with the medical monitor. (iii.f) Within 28 days or 5 half-lives (whichever is longer) before the first dose of any other investigational drug or device [for long half-lives (e.g., > 5 days), enrollment before the 5th half-life requires approval from the medical monitor], (iii.g) Administration by colony-stimulating factors (including granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, or recombinant erythropoietin) within 14 days before study day 1, (iii.h) Has not recovered to ≤ grade 1 from the toxic effects of previous therapy (including previous immunotherapy), and / or complications from previous surgical interventions before the start of therapy [(participants with stable chronic AEs (≤ grade 2) from which recovery is not expected, such as neuropathy and alopecia are exceptions and may be enrolled. Participants with a history of immune-related ocular AEs of any grade are excluded, and participants with a history of grade 3 or higher AEs from previous immunotherapy are excluded from the dose escalation (part 1) of the trial, and participants with a history of grade 3 or higher non-interest AEs may be permitted with approval from the medical monitor)], (iv) Phase 2 - (iv.a) Participants who discontinued anti-PD-L1 therapy for toxicity or other reasons unrelated to toxicity and have since experienced disease progression, (iv.b) Participants who have experienced an objective response (PR / CR) and stopped anti-PD-L1 therapy for maximum benefit, (iv.c) Have achieved a mixed tumor response to previous anti-PD-L1 therapy (such as PR / CR or isolated progressive lesions against SD for other lesions), or have multiple metastases with overall disease progression based on only a single new lesion, (iv.d) Have a disease eligible for potentially curative treatment such as surgical resection [single resection of metastatic lesions against disease progression with previous therapy is not considered curative], (iv.e) Administration of anti-cancer therapy (immunotherapy, chemotherapy, targeted therapy, or hormonal therapy) within 21 days from the first dose of the study treatment, excluding local radiation therapy, (iv.f) Palliative radiotherapy administered within 1 week from the first dose of the investigational treatment, or radiotherapy in the chest area exceeding 30 Gy within 6 months from the first dose of the investigational treatment [Participants must have recovered from all radiation-related toxicities, not require corticosteroids for this purpose, and not have radiation pneumonitis], (iv.g) If the participant has undergone major surgery, they must have recovered appropriately from the toxicity and / or complications due to the intervention before starting the investigational treatment, (iv.h) Treatment-related toxicities related to previous therapy that have not recovered to grade 1 or less (excluding alopecia and anemia not requiring blood transfusion support), except if approved by the medical monitor, (iv.(i) (in accordance with product labels or consent guidelines) a history of immune-related toxicity during previous checkpoint inhibitor therapy, or immune-related toxicity requiring intensive or prolonged immunosuppression to manage (excluding endocrine disorders well controlled with replacement hormones at stable doses, such as hypothyroidism or adrenal insufficiency, or grade 3 rash that resolved with local therapy or asymptomatic lipase elevation that did not require treatment interruption, or uveitis that resolved with steroid drops), (v) having an active autoimmune disease requiring systemic immunosuppression with corticosteroids (>10 mg / day or equivalent) or immunosuppressive drugs within 14 days prior to the first dose of investigational treatment, (vi) chronic systemic corticosteroids (>10 mg / day prednisone or equivalent) [physiological corticosteroid replacement therapy at a dose of prednisone or equivalent >10 mg / day is permitted for adrenal or pituitary insufficiency and in the absence of active autoimmune disease, participants with conditions requiring intermittent use of bronchodilators, inhaled steroids, or local administration of steroid injections may be permitted (e.g., exacerbation of asthma or chronic obstructive pulmonary disease), participants using topical agents may have eye, intra-articular, or intranasal steroids (minimal systemic absorption) and may be hospitalized. Simple courses of corticosteroids for prophylaxis (e.g., contrast dye allergy) or standard premedication related to investigational treatment are permitted], (vii) active infections requiring systemic antibiotics, or antifungal or antiviral treatment within 7 days prior to the first dose of investigational treatment [if the screening test results for SARS-CoV2 infection are positive until test normalization and clinical recovery, the participant should be excluded], (viii) a history of organ transplantation, including allogeneic stem cell transplantation, (ix) evidence of interstitial lung disease or active non-infectious pneumonia, (x) the following known active HBV or HCV infections, or a risk of HBV or HCV reactivation: (x.a) Active hepatitis B infection is defined by positive HBsAg and positive total anti-HBc results. When HBsAg is negative and HBcAb and / or HBsAb are positive, HBV-DNA should be measured. If HBV-DNA is negative, this participant can be enrolled with close monitoring of MTX activity. (x.b) Active hepatitis C is defined by a positive hepatitis C antibody result and a quantitative HCV-RNA result greater than the lower limit of detection of the assay [Participants with positive HCV antibodies are eligible if HCV-RNA is negative. If HCV RNA is undetectable, participants who have received definitive treatment for HCV are allowed]. (xi) Participants known to be HIV positive. (xii) Known active brain or CNS metastases, including carcinomatous meningitis [Previously treated and clinically stable brain or CNS metastases (participants without evidence of progression by imaging and with any neurological symptoms returned to baseline for at least 4 weeks before the first dose of the investigational drug), no evidence of new or enlarging brain metastases or CNS edema, and not requiring steroids for at least 7 days during the study treatment period]. (xiii) Known additional malignancies that are progressive or require active treatment, or other malignancies within 2 years of study enrollment, excluding treated basal cell or squamous cell carcinomas of the skin, superficial bladder cancer, cervical intraepithelial neoplasia, or other non-invasive or indolent malignancies after treatment with curative intent. (xiv) Participants with cardiac dysfunction or clinically significant heart disease: (xiv.a) New York Heart Association class III or IV heart disease, including existing clinically significant ventricular arrhythmias, congestive heart failure, or cardiomyopathy. (xiv.b) Unstable angina. Acute myocardial infarction ≦6 months before study participation. (xiv.c) Other clinically significant heart diseases (e.g., hypertension of grade ≥ 3), (xv) history or presence of abnormal ECG, clinically significant as per the opinion of the principal investigator of the clinical trial [mean QTc interval > 460 milliseconds is excluded (corrected by Fridericia or Bazett formula)], (xvi) women who are pregnant or breastfeeding, (xvii) receiving live vaccines within 30 days from the scheduled start of the test treatment [examples of live vaccines include measles, mumps, rubella, varicella / zoster, yellow fever, rabies, Bacillus Calmette-Guérin, and typhoid vaccine; injectable seasonal influenza vaccines are generally inactivated virus vaccines and are acceptable, while intranasal influenza vaccines are live attenuated vaccines and are not acceptable], (xviii) as determined by the principal investigator of the clinical trial, those that prevent full participation in the test, including administration of the test treatment and attendance at the required test visits, pose significant risks to the participants, or interfere with the interpretation of the test data, are included.

[0268] e. Objectives and Evaluation Items of the Trial The primary, secondary, and exploratory objectives and evaluation items in Phase 1 and Phase 2 include the following (listed in Table 1):

Table 1-1

Table 1-2

Table 1-3

[0269] f. Trial Results The combination regimens were generally well tolerated and consistent with the safety profile of the immune checkpoint inhibitor class. No new toxicities were observed. Immune-related TEAE were reported in two patients on the triplet regimen (one had grade 3 myocarditis and pericardial effusion, and one had an infusion-related reaction), not reported in patients on the doublet regimen, and no grade ≥ 3 infusion-related adverse events were reported in either regimen.

[0270] The overall anti-tumor activity response (by RECIST v1.1) is summarized in Table 2 [disease controls were defined as patients having a complete response, partial response, or stable disease on or after Day 42]:

Table 2

[0271] The objective response rate was 4.8%. The best overall effect in Part 1 was stable disease, and in Part 2, a confirmed and durable partial response was seen in one patient with checkpoint-refractory melanoma (both pembrolizumab and ipilimumab and nivolumab had failed in the first-line setting). This patient completed three-drug therapy for two years and continued to have a durable response at the data cut-off of the first follow-up assessment.

[0272] The best change rate from baseline of the target lesion size (total diameter) of individual patients (n = 21) evaluable for response is shown in Figure 1. The upper limit line indicates the PD (lesion size increased by 20% or more) criterion, and the lower limit line indicates the PR (lesion size decreased by 30% or more) criterion.

[0273] Example 3 Combination Therapy Trial of Anti-PD-1, Anti-TIM-3, and Anti-LAG-3 Active Agents in Head and Neck Squamous Cell Carcinoma (SCCHN) A study is being conducted to evaluate the efficacy of combinations of anti-PD-1, anti-TIM-3, and / or anti-LAG-3 agents in participants with head and neck squamous cell carcinoma (SCCHN). A study population of approximately 164 patients has been selected with treatment-naive recurrent or metastatic PD-L1+ SCCHN.

[0274] This study included three patient cohorts of approximately 54 patients each, randomized 1:1:1 to receive one of three treatments intravenously for up to two years. (Group 1) – INCMGA00012 monotherapy at 500 mg once every four weeks (q4w), +2 placebo control patients; (Group 2) INCMGA00012 at 500 mg q4w, and antibody B at 350 mg once every two weeks (q2w) +1 placebo control patient; and (Group 3) INCMGA00012 at 500 mg q4w, antibody B at 350 mg (q2w), and antibody A at 400 mg (q2w). Patient randomization was stratified by LAG-3 expression status (positive [≥5%] or negative [<5%] tumor proportion score), PD-L1 expression status (combined positive score 1%–19% vs ≥20%), and human papillomavirus p16 status (positive vs negative; oropharyngeal tumors only).

[0275] Patient selection criteria were as follows: (i) being 18 years of age or older, (ii) having an ECOG performance status of 0 or 1, (iii) having recurrent or metastatic SCCHN that: (iii.a) is histologically or cytologically confirmed, (iii.b) has a primary tumor in the oropharynx, oral cavity, hypopharynx, or larynx, and (iii.c) is not suitable for potentially curative surgery or radiotherapy, (iv) having a PD-L1 positive tumor status with a combined positive score of ≥1% as determined by the central laboratory, (v) having ≥1 measurable tumor lesion according to RECIST v1.1, which is a tumor for measurement either (a) that need not have been biopsied or irradiated, or (b) that must show clear evidence of progression from biopsy / radiotherapy.

[0276] The exclusion criteria for patients include the following: (i) previous systemic therapy for metastatic or recurrent SCCHN, (ii) progressive or recurrent disease within 6 months of the end of the last systemic treatment for locally advanced SCCHN, (iii) SCCHN primary tumors of the nasopharynx, paranasal sinuses, or salivary glands, (iv) previous PD-(L)1, LAG-3, TIM-3, or other ICI therapy in any treatment setting, (v) treatment with any other therapy or participation in a clinical trial within 21 days prior to the first study dose, (vi) tumor invasion of major blood vessels with active bleeding, (vii) life expectancy of less than 3 months, (viii) active autoimmune diseases requiring systemic immunosuppression with corticosteroids (>10 mg / day of prednisone or equivalent) or immunosuppressive drugs within 2 years from the first administration of the study treatment, (ix) less than 4 weeks from radiotherapy for CNS or known active CNS metastases and / or carcinomatous meningitis, (x) clinical laboratory values indicating cytopenia, renal / liver impairment, or hypercoagulable state.

[0277] The primary, secondary, and safety objectives and evaluation items for Phase 1 and Phase 2 include the following. Primary efficacy - PFS (from randomization date to the earliest date of disease progression or death), secondary efficacy - (i) objective response (PR+CR), (ii) duration of response (from the earliest date of PR or CR to the earliest date of disease progression or death), (iii) disease control (CR+PR+SD†), and (iv) OS (from randomization date to death from any cause), safety - AE (physical examination, changes in vital signs, blood chemistry) and AEs, reductions, or discontinuations leading to study discontinuation. Radiation oncology evaluations (tomography or magnetic resonance imaging) are performed every 8 weeks.

[0278] Example 4 Combination Therapy Trial of Anti-PD-1, Anti-TIM-3, and Anti-LAG-3 Active Agents in MSI-H Advanced or Metastatic Endometrial Cancer A study is underway to evaluate the efficacy of combination therapy with an anti-PD-1 agent, an anti-TIM-3 agent, and / or an anti-LAG-3 agent in patients with MSI-H advanced or metastatic endometrial cancer who have disease progression during or after platinum-based chemotherapy and have a history of PD-L1 inhibitor therapy. A study population of up to approximately 40 female participants (at least 18 years old) has been selected.

[0279] This trial includes each patient receiving the following treatments intravenously (over 30 minutes) for up to 2 years: INCMGA00012 at 500 mg q4w, antibody B at 350 mg (q2w), and antibody A at 400 mg (q2w).

[0280] The patient selection criteria include the following: (i) female, 18 years of age or older; (ii) histologically confirmed diagnosis of advanced or metastatic endometrial cancer with disease progression during or after treatment with at least one platinum-containing regimen for advanced or metastatic disease; (iii) radiological evidence of disease progression before and after previous PD-(L)1 therapy - (iii.a) participants must have received at least two doses of previous PD-(L)1 therapy; (iii.b) participants may have received PD-(L)1 therapy alone or in combination; (iii.c) disease progression must occur at the time of the first on-treatment scan or the first post-treatment scan and be confirmed by imaging more than 4 weeks after the first evidence of disease progression; note: the baseline scan within the trial may function as the confirmatory scan for progressive disease; (iii.d) participants must have achieved an objective response (CR or PR) to previous PD-L1 therapy and subsequently had disease recurrence; (iv) tumor tissue tested centrally or locally as MSI-H using a PCR assay; (v) must have at least one measurable tumor lesion according to RECIST v1.1; and (vi) ECOG performance status of 0 to 1.

[0281] The exclusion criteria for patients are as follows: (i) a histologically confirmed diagnosis of uterine sarcoma; (ii) having a disease eligible for potentially curative treatment using standard chemotherapy, surgical resection, or chemoradiation therapy; (iii) receiving anticancer therapy within 28 days from the first administration of the investigational treatment, except for local radiation therapy; (iv) toxicity of previous therapy that has not recovered to grade 1 or less (excluding alopecia and anemia not requiring blood transfusion support), except when approved by the medical monitor; (v) managing immune-related toxicity during previous checkpoint inhibitor therapy or severe immune-related toxicity requiring intensive management (e.g., use of infliximab) or prolonged immunosuppression (e.g., less than 6 weeks) (excluding endocrine disorders well controlled with replacement hormones) as recommended (in accordance with the product label or consent guidelines); (vi) previous treatment using a therapy targeting LAG-3 or TIM-3 or lenvatinib; (vii) having achieved a mixed tumor response to previous anti-PD-(L)1 therapy (such as PR / CR or isolated progressive lesions on the background of SD for other lesions), or having multiple metastases achieving overall disease progression based on only a single new lesion; (viii) exclusion criteria values - platelets [<100×109 / L], hemoglobin [<9 g / dL], ANC [<1.5×109 / L], ALT [>2×ULN or >5×ULN for participants with liver metastases], AST [>2×ULN or >5×ULN for participants with liver metastases], total bilirubin [≥1.5×ULN unless conjugated bilirubin ≤ULN (conjugated bilirubin needs to be tested only when total bilirubin exceeds ULN), and when there is no ULN in the medical institution, direct bilirubin must be <40% of total bilirubin], CrCl [<30 mL / min calculated by the Cockcroft-Gault equation (glomerular filtration rate can also be used instead of CrCl)], INR or PT [>1.5×ULN unless on therapeutic anticoagulation], aPTT [>1.(ix) Having an active autoimmune disease that requires systemic immunosuppression with corticosteroids (> 10 mg / day or equivalent) or immunosuppressive drugs within 14 days before the first administration of the investigational treatment; (x) Receiving chronic systemic corticosteroid (> 10 mg / day prednisone or equivalent) administration; (xi) Having an active infection that requires systemic antibiotics, or antifungal or antiviral treatment; (xii) History of organ transplantation, including allogeneic stem cell transplantation; (xiii) Receiving administration of probiotics as the first dose of the investigational drug; (xiv) Known active CNS metastases and / or carcinomatous meningitis; (xv) Known additional malignancy that is progressing or requires active treatment, or history of other malignancy within 2 years from the trial registration, excluding healed basal cell or squamous cell carcinoma of the skin, superficial bladder cancer, cervical intraepithelial neoplasia, or non-invasive or indolent malignancy, or cancer for which the participant has been disease-free for less than 1 year after treatment with intent to cure; (xvi) Known hepatitis B or C (total result of HBsAg positive and anti-HBc positive) or co-infection with HIV, HBV, HCV, or HDV; (xvii) Known hypersensitivity to the investigational drug, excipients, or another monoclonal antibody that cannot be controlled by standard means (e.g., antihistamines and corticosteroids); (xviii) Participants with cardiac dysfunction or clinically significant heart disease; (xix) Women who are pregnant or breastfeeding; (xx) If the participant has undergone major surgery, they must have recovered appropriately from the toxicity and / or complications due to the intervention before starting the investigational treatment; (xxi) Receiving live vaccine inoculation within 28 days from the scheduled start of the investigational treatment; (xxii) Evidence of interstitial lung disease or active non-infectious pneumonia; (xxiii) Current use of prohibited drugs; (xxiv) History of gastrointestinal conditions that may affect oral drug absorption (e.g., inflammatory bowel disease, Crohn's disease, ulcerative colitis) is included.

[0282] The primary, secondary, safety objectives and evaluation items are as follows: (i) defined as the proportion of participants having a complete response (CR) or partial response (PR) according to overall response rate (ORR) RECIST v1.1 (determined by the principal investigator of the clinical trial), (ii) evaluating the safety and tolerability of combined INCMGA00012, (iii) evaluating the measure of clinical activity regarding the combination of INCMGA00012, (iii.a) DOR may be provided when a sufficient number of responders are observed in each treatment group. (iii.b) DCR defined as the proportion of participants having CR, PR or SD (determined by the principal investigator of the clinical trial) as the best response, (iii.c) PFS defined as the time from the first dose of the test treatment to death due to disease progression (defined by the principal investigator of the clinical trial) or any cause, (iii.d) OS defined as the time from the first administration of the test treatment to death due to any cause, (iv) evaluating the PK of INCMGA00012 when administered in combination with other therapies (including C max , T max , C min , and AUC 0-t ), (v) evaluating the PK of each of the other investigational drugs (including C max , T max , C min , and AUC 0-t ) when administered in combination with INCMGA00012, (vi) evaluating the immunogenicity of INCMGA00012 when administered in combination with antibody B and antibody A (the proportion of participants developing anti-drug antibodies (ADA)), (vii) evaluating the relationship between baseline biomarkers and treatment biomarkers of clinical activity (association of blood or tumor analytes), immune cell profile and other related marker levels from baseline with treatment outcomes), are included.

Claims

**Claim 1** A method for treating cancer in a human subject in need thereof, comprising administering to the subject (i) an anti-PD-1 active agent at about 375-500 mg once every three weeks or once every four weeks, (ii) an anti-TIM-3 active agent at about 400-1000 mg once every two weeks, once every three weeks, or once every four weeks, and (iii) an anti-LAG-3 active agent at about 350-750 mg once every two weeks, once every three weeks, or once every four weeks. **Claim 2** The method according to claim 1, comprising administering to the subject (i) an anti-PD-1 active agent at 500 mg once every four weeks, (ii) an anti-TIM-3 active agent at 400 mg once every two weeks, and (iii) an anti-LAG-3 active agent at 350 mg once every two weeks. **Claim 3** The method according to claim 1, comprising administering to the subject at least two of (i) an anti-PD-1 active agent at 375 mg once every three weeks, (ii) an anti-TIM-3 active agent at 500 mg once every three weeks, and (iii) an anti-LAG-3 active agent at 450 mg once every three weeks. **Claim 4** The method according to claim 1, comprising administering to the subject at least two of (i) an anti-PD-1 active agent at 375 mg once every three weeks, (ii) an anti-TIM-3 active agent at 1000 mg once every three weeks, and (iii) an anti-LAG-3 active agent at 750 mg once every three weeks. **Claim 5** The method according to any one of claims 1-4, comprising (a) administering the anti-PD-1 active agent and the anti-TIM-3 active agent to the subject, (b) administering the anti-PD-1 active agent and the anti-LAG-3 active agent to the subject, (c) administering the anti-TIM-3 active agent and the anti-LAG-3 active agent to the subject, or (d) administering the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent to the subject. **Claim 6** A combination therapy comprising at least two of (i) an anti-PD-1 active agent at about 375-500 mg, (ii) an anti-TIM-3 active agent at about 400-1000 mg, and (iii) an anti-LAG-3 active agent at about 350-750 mg. **Claim 7** The combination therapy according to claim 6, comprising at least two of 500 mg of an anti-PD-1 active agent, 400 mg of an anti-TIM-3 active agent, and about 350 mg of an anti-LAG-3 active agent.

8. The combination therapy according to claim 6, comprising at least two of 375 mg of an anti-PD-1 active agent, 500 mg of an anti-TIM-3 active agent, and 450 mg of an anti-LAG-3 active agent.

9. The combination therapy according to claim 6, comprising at least two of 375 mg of an anti-PD-1 active agent, 1000 mg of an anti-TIM-3 active agent, and 750 mg of an anti-LAG-3 active agent.

10. The combination therapy according to any one of claims 6 to 9, wherein the combination therapy comprises (a) the anti-PD-1 active agent and the anti-TIM-3 active agent, (b) the anti-PD-1 active agent and the anti-LAG-3 active agent, (c) the anti-TIM-3 active agent and the anti-LAG-3 active agent, or (d) the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent.

11. A combination therapy for use in the treatment of cancer in a human subject in need thereof, the combination therapy comprising at least two of about 375 to 500 mg of an anti-PD-1 active agent, about 400 to 1000 mg of an anti-TIM-3 active agent, and about 350 mg of an anti-LAG-3 active agent.

12. The combination therapy for use according to claim 11, comprising at least two of 500 mg of an anti-PD-1 active agent, 400 mg of the anti-TIM-3 active agent, and 375 mg of the anti-LAG-3 active agent.

13. The combination therapy for use according to claim 11, comprising at least two of 375 mg of an anti-PD-1 active agent, 500 mg of the anti-TIM-3 active agent, and 450 mg of the anti-LAG-3 active agent.

14. The combination therapy for use according to claim 11, comprising at least two of 375 mg of an anti-PD-1 active agent, 1000 mg of the anti-TIM-3 active agent, and 750 mg of the anti-LAG-3 active agent.

15. The combination therapy according to any one of claims 11 to 14, wherein the combination therapy comprises (a) the anti-PD-1 active agent and the anti-TIM-3 active agent, (b) the anti-PD-1 active agent and the anti-LAG-3 active agent, (c) the anti-TIM-3 active agent and the anti-LAG-3 active agent, or (d) the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent.

16. An anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent for use in the treatment of cancer in a human subject, wherein the anti-PD-1 active agent is administered once every 3 or 4 weeks at a dose of about 375 to 500 mg, the anti-TIM-3 active agent is administered every 2, 3, or 4 weeks at a dose of about 400 to 1000 mg, and the anti-LAG-3 agent is administered once every 2, 3, or 4 weeks at a dose of about 350 to 750 mg.

17. The anti-PD-1 active agent for use according to claim 16, which is for administering 500 mg once every 4 weeks, the anti-TIM-3 active agent is for administering 400 mg once every 2 weeks, and the anti-LAG-3 agent is for administering 350 mg once every 2 weeks.

18. The anti-PD-1 active agent for use according to claim 16, which is administered once every 3 weeks at a dose of 375 mg, the anti-TIM-3 active agent is administered once every 3 weeks at a dose of 500 mg, and the anti-LAG-3 agent is administered once every 3 weeks at a dose of 450 mg.

19. The anti-PD-1 active agent for use according to claim 16, which is administered once every 3 weeks at a dose of 375 mg, the anti-TIM-3 active agent is administered once every 3 weeks at a dose of 1000 mg, and the anti-LAG-3 agent is administered once every 3 weeks at a dose of 750 mg.

20. An anti-PD-1 active agent for use in the treatment of cancer in a human subject, wherein the anti-PD-1 active agent is administered once every 3 or 4 weeks at a dose of about 375 to 500 mg, and is administered simultaneously or sequentially with at least one of (i) an anti-TIM-3 active agent administered once every 2, 3, or 4 weeks at a dose of about 400 to 1000 mg, and (ii) an anti-LAG-3 agent administered once every 2, 3, or 4 weeks at a dose of about 350 to 750 mg.

21. The anti-PD-1 active agent according to claim 20, wherein the anti-PD-1 active agent is administered once every 4 weeks at a dose of 500 mg, and is administered simultaneously or sequentially with at least one of (i) an anti-TIM-3 active agent administered once every 2 weeks at a dose of 400 mg, and (ii) an anti-LAG-3 agent administered once every 2 weeks at a dose of 350 mg.

22. The anti-PD-1 active agent according to claim 20, wherein the anti-PD-1 active agent is administered once every 3 weeks at a dose of 375 mg, and is administered simultaneously or sequentially with at least one of (i) an anti-TIM-3 active agent administered once every 3 weeks at a dose of 500 mg, and (ii) an anti-LAG-3 agent administered once every 3 weeks at a dose of 450 mg.

23. The anti-PD-1 active agent according to claim 20, wherein the anti-PD-1 active agent is administered once every 3 weeks at a dose of 375 mg, and is administered simultaneously or sequentially with at least one of (i) an anti-TIM-3 active agent administered once every 3 weeks at a dose of 1000 mg, and (ii) an anti-LAG-3 agent administered once every 3 weeks at a dose of 750 mg.

24. The anti-PD-1 active agent according to any one of claims 20 to 23, wherein the anti-PD-1 active agent is administered simultaneously or sequentially with (a) the anti-TIM-3 active agent, (b) the anti-LAG-3 active agent, or (c) both the anti-TIM-3 active agent and the anti-LAG-3 active agent.

25. Use of at least two of an anti-PD-1 activating agent, an anti-TIM-3 activating agent, and an anti-LAG-3 activating agent in the treatment of cancer in a human subject, wherein the anti-PD-1 activating agent is administered to the subject once every three or four weeks at a dose of about 375 to 500 mg, the anti-TIM-3 activating agent is administered to the subject once every two, three, or four weeks at a dose of about 400 to 1000 mg, and the anti-LAG-3 activating agent is administered to the subject once every two, three, or four weeks at a dose of about 350 to 750 mg. Use of at least two of an anti-PD-1 activating agent, an anti-TIM-3 activating agent, and an anti-LAG-3 activating agent in the treatment of cancer in a human subject.

26. The use of at least two activating agents according to claim 25, wherein the anti-PD-1 activating agent is administered to the subject once every four weeks at a dose of 500 mg, the anti-TIM-3 activating agent is administered to the subject once every two weeks at a dose of 400 mg, and the anti-LAG-3 activating agent is administered to the subject once every two weeks at a dose of 350 mg.

27. The use of at least two activating agents according to claim 25, wherein the anti-PD-1 activating agent is administered to the subject once every three weeks at a dose of 375 mg, the anti-TIM-3 activating agent is administered to the subject once every three weeks at a dose of 500 mg, and the anti-LAG-3 activating agent is administered to the subject once every three weeks at a dose of 450 mg.

28. The use of at least two activating agents according to claim 25, wherein the anti-PD-1 activating agent is administered to the subject once every three weeks at a dose of 375 mg, the anti-TIM-3 activating agent is administered to the subject once every three weeks at a dose of 1000 mg, and the anti-LAG-3 activating agent is administered to the subject once every three weeks at a dose of 750 mg.

29. Use of at least two of an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent in the treatment of cancer in a human subject, wherein the anti-PD-1 active agent is administered to the subject once every 3 or 4 weeks at a dose of about 375 to 500 mg, the anti-TIM-3 active agent is administered to the subject once every 2, 3, or 4 weeks at a dose of about 400 to 1000 mg, and the anti-LAG-3 active agent is administered to the subject once every 2, 3, or 4 weeks at a dose of about 350 to 750 mg, for the manufacture of one or more medicaments for the treatment of cancer in a human subject, use of at least two of an anti-PD-1 active agent, an anti-TIM-3 active agent, and an anti-LAG-3 active agent.

30. The use of at least two active agents according to claim 29, wherein the anti-PD-1 active agent is administered to the subject once every 4 weeks at a dose of 500 mg, the anti-TIM-3 active agent is administered to the subject once every 2 weeks at a dose of 400 mg, and the anti-LAG-3 active agent is administered to the subject once every 2 weeks at a dose of 350 mg.

31. The use of at least two active agents according to claim 29, wherein the anti-PD-1 active agent is administered to the subject once every 3 weeks at a dose of 375 mg, the anti-TIM-3 active agent is administered to the subject once every 3 weeks at a dose of 500 mg, and the anti-LAG-3 active agent is administered to the subject once every 3 weeks at a dose of 450 mg.

32. The use of at least two active agents according to claim 29, wherein the anti-PD-1 active agent is administered to the subject once every 3 weeks at a dose of 375 mg, the anti-TIM-3 active agent is administered to the subject once every 3 weeks at a dose of 1000 mg, and the anti-LAG-3 active agent is administered to the subject once every 3 weeks at a dose of 750 mg.

33. The use of at least two active agents according to any one of claims 25 to 32, wherein the use comprises (a) the anti-PD-1 active agent and the anti-TIM-3 active agent, (b) the anti-PD-1 active agent and the anti-LAG-3 active agent, (c) the anti-TIM-3 active agent and the anti-LAG-3 active agent, or (d) the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent.

34. A kit for treating cancer in a human subject in need thereof, comprising: (i) an anti-PD-1 active agent of about 375-500 mg, (ii) an anti-TIM-3 active agent of about 400-1000 mg, and (iii) an anti-LAG-3 active agent of about 350-750 mg, at least two of which, and instructions for administering the anti-PD-1 active agent once every three weeks or once every four weeks, and the anti-TIM-3 and anti-LAG-3 active agents once every two weeks, once every three weeks, or once every four weeks.

35. A kit according to claim 34, comprising: (i) at least two of 500 mg of an anti-PD-1 active agent, (ii) 400 mg of an anti-TIM-3 active agent, and (iii) 350 mg of an anti-LAG-3 active agent, and instructions for administering the anti-PD-1 active agent once every four weeks and the anti-TIM-3 and anti-LAG-3 active agents once every two weeks.

36. A kit according to claim 34, comprising at least two of (i) 375 mg of an anti-PD-1 active agent, (ii) 500 mg of an anti-TIM-3 active agent, and (iii) 450 mg of an anti-LAG-3 active agent, and instructions for administering the anti-PD-1, anti-TIM-3, and anti-LAG-3 active agents once every three weeks.

37. A kit according to claim 34, comprising at least two of (i) 375 mg of an anti-PD-1 active agent, (ii) 1000 mg of an anti-TIM-3 active agent, and (iii) 750 mg of an anti-LAG-3 active agent, and instructions for administering the anti-PD-1, anti-TIM-3, and anti-LAG-3 active agents once every three weeks.

38. The kit according to any one of claims 34-37, wherein the kit comprises (a) the anti-PD-1 active agent and the anti-TIM-3 active agent, (b) the anti-PD-1 active agent and the anti-LAG-3 active agent, (c) the anti-TIM-3 active agent and the anti-LAG-3 active agent, or (d) the anti-PD-1 active agent, the anti-TIM-3 active agent, and the anti-LAG-3 active agent.

39. The anti-PD-1 active agent comprises an anti-PD-1 antibody or an anti-PD-1 binding fragment thereof, and the PD-1 binding fragment is (i) a heavy chain variable domain (VH) comprising a CDRH1 domain having the amino acid sequence SYWMN (SEQ ID NO: 1), a CDRH2 domain having the amino acid sequence VIHPSDSETWLDQKFK (SEQ ID NO: 2), and a CDRH3 domain having the amino acid sequence EHYGTSPFAY (SEQ ID NO: 3); (ii) a method, combination therapy, use, or kit according to any one of claims 1 to 38, comprising a light chain variable domain (VL) comprising a CDRL1 domain having the amino acid sequence RASESVDNYGM SF MNW (SEQ ID NO: 4), a CDRL2 domain having the amino acid sequence AASNQGS (SEQ ID NO: 5), and a CDRL3 domain having the amino acid sequence QQSK EV PY T (SEQ ID NO: 6).

40. The heavy chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKKPGASVKVSCKASGYSFTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDQKFKDRVTITVDKSTSTAYMELSSLRS EDTAVYYCAREHYGTSPFAYWGQGTLVTVSS (SEQ ID NO: 7). A method, combination therapy, use, or kit according to claim 39.

41. The light chain variable domain of the anti-PD-1 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASESVDNYGM SF MNW FQQKPGQPPKLLIH AASNQGSGVP SRFSGSGSGTDFTLTISSL EPEDFAVYFCQQSK EV PY TFGGG TKVEIK (SEQ ID NO: 8). A method, combination therapy, use, or kit according to any one of claims 39 to 40.

42. The method, combination therapy, use, or kit according to any one of claims 39 to 41, wherein the anti-PD-1 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQLVQSGAEVKPGASVKVSCKASGYTSYWMNWVRQAPGQGLEWIGVIHPSDSETWLDDQKFKDDRVTITVDKSTSTAYMELSSLRSEDTAYYCAREHYGTSPFAYWGQGTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNYVDGVEVHNAKTKPREEQFNSTYRVVSSVLTVLHQDWLNGKEYKCKVSNKGLPSSEIEKTISKAKGQPREPQVYTLPPSQEEMTKNPVSLTCLVKGFYPSDIAVEWESNGQPENNYYKTPPPVLDSDGSFFLYSRLLVDKSRWQEGNVFSCSSMHEALHNHYTQKSLSLSLG (SEQ ID NO: 9).

43. The method, combination therapy, use, or kit according to any one of claims 39 to 42, wherein the anti-PD-1 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASSEVDNYGMSFMNWFRQQKPGQPPKLLIHAA SNQGGSGPSRFSSGSGSGTDFTLTISSLPEDFAVYFCQQSKEVPYTFGGGTVEIKRTVAAPS VFIFPPSEQLKSGTASVVCLNNFYPREAKVQWKVDNALQSGSQESVT EQDSKDS TYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

44. The method, combination therapy, use, or kit according to any one of claims 39 to 43, wherein the anti-PD-1 antibody or a fragment thereof comprises an Fc region of the IgG1, IgG2, IgG3, or IgG4 isotype, optionally wherein the Fc region is of the IgG4 isotype, and the antibody comprises a hinge domain of the IgG4 isotype that contains a stabilizing mutation.

45. The anti-PD-1 antibody or a fragment thereof is (A) one or more amino acid modifications that reduce the affinity of the variant Fc region for FcγR, wherein the one or more modifications that reduce the affinity of the variant Fc region for FcγR include the substitution of L234A, L235A, or L234A + L235A, and the numbering is according to the EU index like Kabat, one or more amino acid modifications, and / or (B) one or more amino acid modifications that enhance the serum half-life of the variant Fc region, wherein the one or more modifications that enhance the serum half-life of the variant Fc region include the substitution of M252Y, M252Y + S254T, M252Y + T256E, M252Y + S254T + T256E, or K288D + H435K, and the numbering is according to the EU index like Kabat, one or more amino acid modifications, and having a variant Fc region, the method, combination therapy, use, or kit according to any one of claims 39 to 44.

46. The method, combination therapy, use, or kit according to any one of claims 39 to 45, wherein the anti-PD-1 antibody is retifanlimab.

47. The method, combination therapy, use, or kit according to any one of claims 1 to 46, wherein the anti-PD-1 active agent is administered intravenously.

48. The anti-TIM-3 active agent comprises an anti-TIM-3 antibody or a TIM-3 binding fragment thereof, and the TIM-3 binding fragment is (i) a heavy chain variable domain (VH) comprising a CDRH1 domain having the amino acid sequence RQNAWS (SEQ ID NO: 11), a CDRH2 domain having the amino acid sequence WVSASGSGGSTY (SEQ ID NO: 12), and a CDRH3 domain having the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 13), and (ii) The method, combination therapy, use, or kit according to any one of claims 1 to 47, comprising a light chain variable domain (VL) containing a CDRL1 domain having the amino acid sequence RASQSVSSYLA (SEQ ID NO: 14), a CDRL2 domain having the amino acid sequence DASNRAT (SEQ ID NO: 15), and a CDRL3 domain having the amino acid sequence QQYGSSPLT (SEQ ID NO: 16):

49. The method, combination therapy, use, or kit according to claim 48, wherein the heavy chain variable domain of the anti-TIM-3 antibody or its fragment comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVRRAPGKGL EWVSAISGSGGSTYYADS VKGRFTISRDN SKNTLYLQMNSLR AEDTAVYYCAKGGDYGGNYFDYWGQGTVTVSS (SEQ ID NO: 17).

50. The method, combination therapy, use, or kit according to any one of claims 48 to 49, wherein the light chain variable domain of the anti-TIM-3 antibody or its fragment comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI YDASNRATGIPASF SGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK (SEQ ID NO: 18).

51. The method, combination therapy, use, or kit according to any one of claims 48 to 50, wherein the anti-TIM-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EVQLVESGGGLVQPGGSLRLSCAASGFTFRQNAWSWVVRRAPGKGL EWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGDYGGNYFDYWGQGTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVSDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLLSLSPG (SEQ ID NO: 19).

52. The method, combination therapy, use, or kit according to any one of claims 48 to 51, wherein the anti-TIM-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPATLSLSPGERATLSCRASQSVS SYLAWYQQKPGQAPRLLIYDASNRATGIPASFGSGSGSTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTVKEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGSQESVTEQDSKDS TYSLSSLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).

53. The method, combination therapy, use, or kit according to any one of claims 48 to 52, wherein the anti-TIM-3 antibody or fragment thereof comprises an Fc region of human IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype.

54. The method, combination therapy, use, or kit according to any one of claims 48 to 53, wherein the anti-TIM-3 antibody or fragment thereof comprises an Fc region of human IgG1 isotype, and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system or an N297Q mutation numbered according to the EU numbering system.

55. The method, combination therapy, use, or kit according to any one of claims 48 to 54, wherein the anti-TIM-3 antibody or fragment thereof comprises an Fc region of human IgG4 isotype, and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system or an N297Q mutation numbered according to the EU numbering system.

56. The method, combination therapy, use, or kit according to any one of claims 48 to 55, wherein the anti-TIM-3 antibody or fragment thereof is antagonist to human TIM-3, and optionally, the anti-TIM-3 antibody or fragment thereof inactivates, reduces, or inhibits the activity of human TIM-3, and optionally, the anti-TIM-3 antibody or fragment thereof inhibits the binding of human TIM-3 to phosphatidylserine.

57. The method, combination therapy, use, or kit according to any one of claims 48 to 56, wherein the anti-TIM-3 antibody is antibody A.

58. The method, combination therapy, use, or kit according to any one of claims 1 to 57, wherein the anti-TIM-3 active agent is administered intravenously.

59. The anti-LAG-3 active agent comprises an anti-LAG-3 antibody or an anti-LAG-3 binding fragment thereof, and the anti-LAG-3 binding fragment i) CDRH1 domain having the amino acid sequence DT YI Ｈ a heavy chain variable domain (VH) comprising a CDRH2 domain having the amino acid sequence EI DPA NDT KY DPK FQG (SEQ ID NO: 22) and a CDRH3 domain having the amino acid sequence YYY KY DVG GFD Y (SEQ ID NO: 23), and ii) a light chain variable domain (VL) comprising a CDRL1 domain having the amino acid sequence SVSSSSISSSNLH (SEQ ID NO: 24), a CDRL2 domain having the amino acid sequence GTSNLAS (SEQ ID NO: 25), and a CDRL3 domain having the amino acid sequence QQWSSYPFT (SEQ ID NO: 26). The method, combination therapy, use, or kit according to any one of claims 1 to 58.

60. The method, combination therapy, use, or kit according to claim 59, wherein the heavy chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKPGASVKVSCKASGFNIKDTYIHWVQRQAPGQGLEWMGEIDPANDNTKYDPKFQG RVTITADTSTSTVYMELSSLRS EDTAVYYCATYYYKYDVGGFDYWGQGTVTVSS (SEQ ID NO: 27).

61. The method, combination therapy, use, or kit according to any one of claims 59 to 60, wherein the light chain variable domain of the anti-LAG-3 antibody or fragment thereof comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLS LSPSGERATLSCSVSSSISSSNLHWYQQKPGQAPRLLIYGT SNLASGIPDRFSGSGSGTDFTLTISRLPE DFAVYYCQQWSSYPFTFGQGTVEIK (SEQ ID NO: 28).

62. The method, combination therapy, use, or kit according to any one of claims 59 to 61, wherein the anti-LAG-3 antibody or fragment thereof comprises a heavy chain (HC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to QVQMVQSGAEVKKPGASVKVSCKASGFNIKDTYIHWVRQAPGQGLEWMGEIDPANDNTKYDPKFQGRVTITADTSTSTVYMELSSLRS EDTAVYYCATYYYKYDVGGFDYWGQGTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK SCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPERKFNWYVDGVEVHNAKTKPREEQYASTYRVVSSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSSVMHEALHNHYTQKSLLSLSPG (SEQ ID NO: 29).

63. The method, combination therapy, use, or kit according to any one of claims 59 to 62, wherein the anti-LAG-3 antibody or fragment thereof comprises a light chain (LC) sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to EIVLTQSPGTLSLSPEGERALSCSSSSISSSNLHWYQQKPGQAPRLLIYGTSNLASGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQWSSYPFTFGQGTVEIKRTVAAPSVFIFFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGSQESVTEQDSKDS TYSLSSLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).

64. The method, combination therapy, use, or kit according to any one of claims 59 to 63, wherein the anti-LAG-3 antibody or fragment thereof comprises an Fc region of human IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2 isotype.

65. The method, combination therapy, use, or kit according to any one of claims 59 to 64, wherein the anti-LAG-3 antibody or fragment thereof comprises an Fc region of human IgG1 isotype, and the amino acid sequence of the IgG1 heavy chain constant region comprises an N297A mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system.

66. The method, combination therapy, use, or kit according to any one of claims 59 to 65, wherein the anti-LAG-3 antibody or fragment thereof comprises an Fc region of human IgG4 isotype, and the amino acid sequence of the IgG4 heavy chain constant region comprises an S228P mutation numbered according to the EU numbering system, or an N297Q mutation numbered according to the EU numbering system.

67. The method, combination therapy, use, or kit according to any one of claims 59 to 66, wherein the anti-LAG-3 antibody or fragment thereof is antagonistic to human LAG-3, and optionally, the anti-LAG-3 antibody or fragment thereof inactivates, reduces, or inhibits the activity of human LAG-3, and optionally, the anti-LAG-3 antibody or fragment thereof inhibits the binding of human LAG-3 to MHC class II.

68. The method, combination therapy, use, or kit according to any one of claims 59 to 67, wherein the anti-LAG-3 antibody is antibody B.

69. The method, combination therapy, use, or kit according to any one of claims 1 to 68, wherein the anti-LAG-3 active agent is administered intravenously.

70. The method, combination therapy, use, or kit according to any one of claims 1 to 69, wherein the anti-PD-1 active agent is administered before the anti-TIM-3 active agent and the anti-LAG-3 active agent, and optionally, (a) the anti-TIM-3 composition is administered before the anti-LAG-3 composition, (b) the anti-LAG-3 composition is administered before the anti-TIM-3 composition, or (c) the anti-TIM-3 composition and the anti-LAG-3 composition are administered simultaneously.

71. The anti-PD-1 active agent is administered after the anti-TIM-3 active agent and the anti-LAG-3 active agent, and optionally, (a) the anti-TIM-3 composition is administered before the anti-LAG-3 composition, (b) the anti-LAG-3 composition is administered before the anti-TIM-3 composition, or (c) the anti-TIM-3 composition and the anti-LAG-3 composition are administered simultaneously, the method, combination therapy, use, or kit according to any one of claims 1 to 70.

72. The anti-PD-1 active agent is administered in a pharmaceutical composition comprising acetate, sucrose, polysorbate 80 (“PS80”), and water, has a pH of about 4.0 to about 6.5, and optionally, the concentration of the anti-PD-1 agent in the pharmaceutical composition is about 10 mg / mL to about 100 mg / mL, the method, combination therapy, use, or kit according to any one of claims 1 to 71.

73. The anti-TIM-3 active agent is administered in a pharmaceutical composition comprising sodium citrate, sucrose, arginine, and polysorbate 80, has a pH of 6.0, and optionally, the concentration of the anti-TIM-3 agent in the pharmaceutical composition is about 50 mg / mL, the method, combination therapy, use, or kit according to any one of claims 1 to 72.

74. The anti-LAG-3 active agent is administered in a pharmaceutical composition comprising sodium acetate, trehalose, and polysorbate 80, has a pH of 5.5, and optionally, the concentration of the anti-LAG-3 agent in the pharmaceutical composition is about 50 mg / mL, the method, combination therapy, use, or kit according to any one of claims 1 to 73.

75. The cancer comprises a tumor, and optionally, the tumor is a locally advanced tumor, a metastatic solid tumor, or a combination thereof, the method, combination therapy, use, or kit according to any one of claims 1 to 74.

76. The cancer comprises a tumor in which a PD-1 inhibitor is indicated, the method, combination therapy, use, or kit according to any one of claims 1 to 75.

77. The method, combination therapy, use, or kit according to any one of claims 75-76, wherein the subject has been pre-treated with at least one anti-PD-1 / anti-PDL-1 therapy, optionally, the subject has cancer that has failed at least one anti-PD-1 / PDL-1 inhibitor therapy, and optionally, the subject has cancer that continues to progress during the previous anti-PD-1 / anti-PDL-1 therapy.

78. The method, combination therapy, use, or kit according to any one of claims 75-77, wherein the tumors have acquired resistance to anti-PD-1 therapy, or have innate resistance to anti-PD-1 therapy, or combinations thereof.

79. The method, combination therapy, use, or kit according to any one of claims 75-78, wherein the tumors have a minimum LAG-3 expression of 5% or more of LAG-3 positive immune cells (e.g., lymphocytes and macrophages) relative to all nucleated cells within the tumor region, as shown by immunohistochemical assay.

80. The method, combination therapy, use, or kit according to any one of claims 1-79, wherein the cancer is melanoma, optionally unresectable and / or metastatic melanoma.

81. The method, combination therapy, use, or kit according to any one of claims 1-79, wherein the cancer is squamous cell carcinoma of the head and neck (SCCHN), optionally recurrent or metastatic PD-L1+ SCCHN.

82. The method, combination therapy, use, or kit according to any one of claims 1-79, wherein the cancer is endometrial cancer, optionally MSI-H advanced cancer or metastatic endometrial cancer, and optionally has evidence of disease progression inside and outside platinum-based chemotherapy and previous PD-L1 inhibitory therapy.

83. The method, combination therapy, use, or kit according to any one of claims 1-82, wherein the administration or treatment results in at least one treatment effect selected from reduction in tumor size, reduction in the number of metastatic lesions over time, complete response, partial response, and stable disease, according to the RECIST v1.1 criteria.

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