Methods of treating tumor

Abstract

To provide a method for treating a subject afflicted with a tumor, for example, a tumor derived from non-small cell lung cancer (NSCLC).SOLUTION: A method comprises administering to the subject a combination of (a) a chemotherapy agent, (b) an anti-PD-1 antibody or an anti-PD-L1 antibody, and (c) an anti-CTLA-4 antibody, where the chemotherapy agent is administered for a period of time that is less than the standard.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This PCT application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 749,393, filed October 23, 2018, the entire contents of which are incorporated herein by reference.

[0002] Technical Field The present invention provides methods for treating a subject suffering from a tumor using immunotherapy in combination with chemotherapy. [Background technology]

[0003] Background technology Immunotherapy has recently demonstrated clinical efficacy in several cancer types, including melanoma and hormone-refractory prostate cancer. Tumors can modulate and evade host immune responses through numerous mechanisms, including downregulation of tumor-specific antigen expression and presentation, secretion of anti-inflammatory cytokines, and upregulation of inhibitory ligands. T cell checkpoint regulators, such as CTLA-4 and programmed cell death-1 (PD-1, CD279), are cell surface molecules that, when engaged by their cognate ligands, induce signaling cascades that downregulate T cell activation and proliferation.

[0004] PD-1 is an important immune checkpoint receptor expressed by activated T cells and B cells and mediates immunosuppression. PD-1 is a member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1, programmed cell death ligand-1 (PD-L1) and programmed cell death ligand-2 (PD-L2), have been identified and are expressed on antigen-presenting cells and in many human cancers. Binding to PD-1 has been shown to downregulate T cell activation and cytokine secretion. Inhibition of the PD-1 / PD-L1 interaction mediates potent antitumor activity in preclinical models (U.S. Patent Nos. 8,008,449 and 7,943,743), and the use of antibody inhibitors of the PD-1 / PD-L1 interaction to treat cancer has entered clinical trials (Brahmer et al., 2010; Topalian et al., 2012a; Topalian et al., 2014; Hamid et al., 2013; Brahmer et al., 2012; Flies et al., 2011; Pardoll, 2012; Hamid and Carvajal, 2013).

[0005] Nivolumab (previously known as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby preventing downregulation of antitumor T cell function (U.S. Patent No. 8,008,449; Wang et al., 2014). Nivolumab has demonstrated activity in a variety of advanced solid tumors, including renal cell carcinoma (renal adenocarcinoma or adrenal nephroma), melanoma, and non-small cell lung cancer (NSCLC) (Topalian et al., 2012a; Topalian et al., 2014; Drake et al., 2013; WO 2013 / 173223).

[0006] Combining immunotherapeutic agents with different mechanisms of action offers the potential for synergistic responses. PD-1 and CTLA-4 are both co-inhibitory molecules, but evidence suggests they use different mechanisms to limit T cell activation. Ipilimumab and anti-CTLA-4 antibodies have been shown to enhance the anticancer activity of nivolumab.

[0007] However, there is a need to further improve the effectiveness of anti-PD-1 or anti-PD-L1 therapy. Described herein are methods of administering a combination of an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and platinum-based doublet chemotherapy. Summary of the Invention

[0008] Summary of the Invention Certain aspects of the present invention are directed to a method of treating a tumor in a subject in need thereof, comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) The method further comprises, after (1), a post-induction phase comprising administering to the subject an antibody that specifically binds to PD-1 (an "anti-PD-1 antibody") or an antigen-binding portion thereof, or an antibody that specifically binds to PD-L1 (an "anti-PD-L1 antibody") or an antigen-binding portion thereof. The present invention can provide the following aspects. [Section 1] 1. A method for treating a tumor in a subject in need thereof, comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) (1) is followed by an induction period that includes administering to the subject an antibody that specifically binds to PD-1 (an "anti-PD-1 antibody") or an antigen-binding portion thereof, or an antibody that specifically binds to PD-L1 (an "anti-PD-L1 antibody") or an antigen-binding portion thereof. A method comprising: [Section 2] A method of treating a tumor in a subject in need thereof, comprising administering to the subject an anti-PD-1 antibody or an anti-PD-L1 antibody, wherein prior to administering the anti-PD-1 antibody or anti-PD-L1 antibody, the subject undergoes an induction phase comprising chemotherapy for a duration that is shorter than the standard duration of the chemotherapeutic agent. [Section 3] Item 3. The method of paragraph 1 or 2, wherein the induction phase further comprises administering an anti-PD-1 antibody or an anti-PD-L1 antibody. [Section 4] The method of any one of paragraphs 1 to 3, further comprising administering an antibody that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody") or an antigen-binding portion thereof. [Section 5] The method of paragraph 4, wherein the anti-CTLA-4 antibody is administered during the induction phase. [Section 6] The method of paragraph 4 or 5, wherein the anti-CTLA-4 antibody is administered after the induction period. [Section 7] The method of any one of paragraphs 4 to 6, wherein the anti-CTLA-4 antibody is administered before or after the anti-PD-1 antibody or anti-PD-L1 antibody. [Section 8] The method of any one of paragraphs 4 to 7, wherein the anti-CTLA-4 antibody is administered after chemotherapy. [Section 9] The method of paragraph 3, wherein the anti-CTLA-4 antibody is administered simultaneously with the anti-PD-1 antibody or anti-PD-L1 antibody. [Section 10] The method of any one of paragraphs 1 to 9, wherein the chemotherapy agent is administered for less than 10 cycles, less than 9 cycles, less than 8 cycles, less than 7 cycles, less than 6 cycles, less than 5 cycles, less than 4 cycles, or less than 3 cycles. [Section 11] 11. The method of any one of paragraphs 1 to 10, wherein the chemotherapy agent is administered in 5 cycles, 4 cycles, 3 cycles, 2 cycles or 1 cycle. [Section 12] 12. The method of any one of paragraphs 1 to 11, wherein the chemotherapy agent is administered for less than 5 cycles. [Section 13] The method of paragraph 12, wherein the chemotherapy agent is administered in less than four cycles. [Section 14] 14. The method of paragraph 13, wherein the chemotherapy agent is administered in less than three cycles. [Section 15] The method of paragraph 14, wherein the chemotherapy agent is administered in less than two cycles. [Section 16] 16. The method of any one of paragraphs 1 to 15, wherein the chemotherapy agent is administered in a maximum of two cycles. [Section 17] 17. The method of any one of paragraphs 1 to 16, wherein the chemotherapy comprises standard therapy. [Section 18] 18. The method of any one of paragraphs 1 to 17, wherein the chemotherapeutic agent comprises an alkylating agent, an antimetabolite, an anti-microtubule agent, a topoisomerase inhibitor, a cytotoxic antibiotic, or any combination thereof. [Section 19] 19. The method of any one of paragraphs 1 to 18, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent. [Section 20] 20. The method of any one of paragraphs 1 to 19, wherein the chemotherapeutic agent comprises cisplatin, oxaliplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, or any combination thereof. [Section 21] 21. The method of any one of paragraphs 1 to 20, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and a second agent. [Section 22] 22. The method of any one of paragraphs 1 to 21, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and paclitaxel. [Section 23] 22. The method of any one of paragraphs 1 to 21, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and pemetrexed. [Section 24] 23. The method of any one of paragraphs 1 to 22, wherein the chemotherapeutic agent comprises carboplatin and paclitaxel. [Section 25] 24. The method of any one of paragraphs 1 to 21 and 23, wherein the chemotherapeutic agent comprises carboplatin and pemetrexed. [Section 26] 24. The method of any one of paragraphs 1 to 21 and 23, wherein the chemotherapeutic agent comprises cisplatin and pemetrexed. [Section 27] 27. The method of any one of paragraphs 1 to 26, wherein the chemotherapeutic agent is a histology-based chemotherapeutic agent. [Section 28] 28. The method of any one of paragraphs 1 to 27, wherein the chemotherapeutic agent is administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, or about once every 6 weeks. [Section 29] 29. The method of any one of paragraphs 10 to 28, wherein each cycle is 3 weeks. [Section 30] 30. The method of paragraph 29, wherein the chemotherapy agent is administered on day 1 of each three-week cycle. [Section 31] Chemotherapy consisted of carboplatin AUC 6 and paclitaxel 200 mg / m on any one day of a 3-week cycle. 2 31. The method of any one of paragraphs 1 to 30, comprising administering [Section 32] Chemotherapy consisted of carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m 2 31. The method of any one of paragraphs 1 to 30, comprising administering [Section 33] Chemotherapy was cisplatin 75mg / m 2 and pemetrexed 500 mg / m 2 31. The method of any one of paragraphs 1 to 30, comprising administering [Section 34] 34. The method of any one of paragraphs 1 to 33, wherein the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. [Section 35] 35. The method of any one of paragraphs 1 to 34, wherein the anti-PD-1 antibody binds to the same epitope as nivolumab. [Section 36] 36. The method of any one of paragraphs 1 to 35, wherein the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. [Section 37] 37. The method of any one of paragraphs 1 to 36, wherein the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype. [Section 38] 38. The method of any one of paragraphs 1 to 37, wherein the anti-PD-1 antibody is nivolumab. [Section 39] 38. The method of any one of paragraphs 1 to 37, wherein the anti-PD-1 antibody is pembrolizumab. [Section 40] 40. The method of any one of paragraphs 1 to 39, wherein the anti-PD-1 antibody is administered at a dose range of 0.1 mg to 10.0 mg per kg of body weight once every 2 weeks, 3 weeks, or 4 weeks. [Section 41] 41. The method of any one of paragraphs 1 to 40, wherein the anti-PD-1 antibody is administered at a dose of 3 mg, 5 mg, or 10 mg per kg of body weight once every three weeks. [Section 42] 40. The method of any one of paragraphs 1 to 39, wherein the anti-PD-1 antibody is administered at a fixed dose. [Section 43] 43. The method of clause 42, wherein the anti-PD-1 antibody is administered at a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg. [Section 44] 44. The method of paragraph 42 or 43, wherein the anti-PD-1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, or about every four weeks. [Section 45] 45. The method of any one of paragraphs 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks. [Section 46] 45. The method of any one of paragraphs 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 240 mg once about every two weeks. [Section 47] 45. The method of any one of paragraphs 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 480 mg once about every four weeks. [Section 48] 34. The method of any one of paragraphs 1 to 33, wherein the anti-PD-L1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. [Section 49] 49. The method of any one of paragraphs 1 to 33 and 48, wherein the anti-PD-L1 antibody comprises a heavy chain constant region of a human IgG1 isotype. [Section 50] 50. The method of any one of paragraphs 1 to 33, 48 and 49, wherein the anti-PD-L1 antibody cross-competes with an antibody selected from atezolizumab, durvalumab and avelumab for binding to human PD-L1. [Section 51] The method of any one of paragraphs 1 to 33 and 48 to 50, wherein the anti-PD-L1 antibody binds to the same epitope on human PD-L1 as an antibody selected from atezolizumab, durvalumab, and avelumab. [Section 52] The method of any one of paragraphs 33 and 48 to 51, wherein the anti-PD-L1 antibody is atezolizumab, durvalumab, or avelumab. [Section 53] The method of any one of paragraphs 1 to 33 and 48 to 52, wherein the anti-PD-L1 antibody is administered at a dose ranging from 0.1 mg / kg to 15.0 mg / kg of body weight once every 2 weeks, 3 weeks, or 4 weeks. [Section 54] 54. The method of any one of paragraphs 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered at a dose of 3 mg or 5 mg per kg of body weight once every two weeks. [Section 55] 54. The method of any one of paragraphs 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered at a dose of 10 mg / kg body weight once every three weeks. [Section 56] 54. The method of any one of paragraphs 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered at a fixed dose. [Section 57] 57. The method of clause 56, wherein the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, at least about 1400 mg, or at least about 1500 mg. [Section 58] 58. The method of paragraph 56 or 57, wherein the anti-PD-L1 antibody is administered as a fixed dose once every week, every two weeks, every three weeks, or every four weeks. [Section 59] 59. The method of any one of paragraphs 1 to 33, 48 to 53, 57 and 58, wherein the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg once about every three weeks. [Section 60] 60. The method of any one of paragraphs 1 to 59, wherein the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or portion thereof. [Section 61] 61. The method of any one of paragraphs 1 to 60, wherein the anti-CTLA-4 antibody comprises a heavy chain constant region of the human IgG1 isotype. [Section 62] 62. The method of any one of paragraphs 1 to 61, wherein the anti-CTLA-4 antibody is ipilimumab. [Section 63] 63. The method of any one of paragraphs 1 to 62, wherein the anti-CTLA-4 antibody is tremelimumab. [Section 64] 64. The method of any one of paragraphs 1 to 63, wherein the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4. [Section 65] 65. The method of any one of paragraphs 1 to 64, wherein the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg to at least about 10.0 mg per kg of body weight about once every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks. [Section 66] 66. The method of any one of paragraphs 1 to 65, wherein the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. [Section 67] 66. The method of any one of paragraphs 1 to 65, wherein the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg of body weight once about every 12 weeks. [Section 68] 68. The method of any one of paragraphs 1 to 67, wherein the anti-CTLA-4 antibody is administered in a fixed dose. [Section 69] (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; and (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight once about every 6 weeks; Item 69. The method of any one of items 1 to 68. [Section 70] (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents were carboplatin AUC 6 and paclitaxel 200 mg / m 2 administered on day 1 of an approximately 3-week cycle, Item 69. The method of any one of items 1 to 68. [Section 71] (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents were carboplatin AUC 5 and pemetrexed 500 mg / m 2 administered on day 1 of an approximately 3-week cycle, Item 70. The method of any one of items 1 to 69. [Section 72] (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents were carboplatin AUC 6 and pemetrexed 500 mg / m 2 administered on day 1 of an approximately 3-week cycle, Item 70. The method of any one of items 1 to 69. [Section 73] (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agent is cisplatin 75 mg / m 2 and pemetrexed 500 mg / m 2 administered on day 1 of an approximately 3-week cycle, Item 70. The method of any one of items 1 to 69. [Section 74] 74. The method of any one of paragraphs 70 to 73, wherein the chemotherapy agent is administered in fewer than three cycles. [Section 75] 75. The method of any one of paragraphs 70 to 74, wherein the chemotherapy agent is administered over two cycles. [Section 76] 76. The method of any one of paragraphs 1 to 75, wherein the subject exhibits a progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. [Section 77] 77. The method of any one of paragraphs 1 to 76, wherein the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. [Section 78] 78. The method of any one of paragraphs 1 to 77, wherein the subject exhibits a response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%. [Section 79] 79. The method of any one of paragraphs 1 to 78, wherein the tumor has a high tumor mutation burden (TMB) status. [Section 80] 80. The method of claim 79, wherein the TMB status is determined by sequencing nucleic acid in the tumor and identifying genomic alterations in the sequenced nucleic acid. [Section 81] Genome modification, (i) one or more somatic mutations; (ii) one or more nonsynonymous mutations; (iii) one or more missense mutations; (iv) one or more alterations selected from the group consisting of base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and combinations thereof; or (v) Any combination of (i) to (iv) 81. A composition for use in the method according to claim 80, comprising [Section 82] High TMB is at least 210, at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 82. The composition for use in the method of any one of paragraphs 79 to 81, having a score of at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495 or at least 500. [Section 83] 83. The composition for use in the method of any one of paragraphs 79 to 82, wherein the subject's TMB status is compared to a control TMB value, wherein the subject's TMB status is within the highest quantile of the control TMB values, or the subject's TMB status is within the highest tertile of the control TMB values. [Section 84] 84. The composition for use in the method of any one of paragraphs 79 to 83, wherein the biological sample comprises a tumor tissue biopsy, a liquid biopsy, blood, serum, plasma, exoRNA, circulating tumor cells, ctDNA, cfDNA, or any combination thereof. [Section 85] TMB status is (i) genome sequencing; (ii) exome sequencing; (iii) genomic profiling, or (iv) Any combination of (i) to (iii) 85. A composition for use in the method of any one of paragraphs 79 to 84, wherein the composition is determined by: [Section 86] ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PDCD1LG2, RBM1 0, STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGFRA, RET, STK1 1, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTOR, SUFU, AKT1 BRD4, CREBBP, FANCF, GID4(C17orf39), KAT6A(MYST3), MRE11A, PDK1, RNF4 3. SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B, ROS1, TAF1. AKT3, BTG1, CRLF2, FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR, TBX3, ALK, BT K, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER1(FAM123B). C11orf30(EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1T1, TERT(プロモAPC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R1, SDHA, TET2 AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2, SDHB, TGFBR2 ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PLCG2, SDHC, TNFAIP3, ARFRP1 CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF14, ARID1A, CC ND2, DAXX, FGF23, GSK3B, KMT2A(MLL), NF1, POLD1, SETD2, TOP1, ARID1B, CC ND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, TOP2A, ARID2, CCNE 1, DICER1, FGF4, HGF, KMT2D(MLL2), NFE2L2, PPP2R1A, SLIT2, TP53, ASXL1 CD274, DNMT3A, FGF6, HNF1A, KRAS, NFKBIA, PRDM1, SMAD2, TSC1, ATM, CD79A;DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR, ATRX, CDC7 3, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARCB1, VEGFA, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WISP3, AXL, C DK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BARD1, CDKN 1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BCL2L1, CDK N2A, ERG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, CDKN2C, 86. The composition of claim 85, comprising one or more genes selected from the group consisting of ESR1, FUBP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and combinations thereof. [Section 87] (i) the tumor comprises non-small cell carcinoma; (ii) the tumor is relapsed or refractory after at least one prior choice of therapy for treating the tumor; or (iii) Both (i) and (ii); 87. A composition for use in a method according to any one of paragraphs 79 to 86. [Section 88] Genomic profiles are available in FOUNDATIONONE® CDX (商標) 86. The composition of claim 85, comprising: [Section 89] 89. The composition for use in the method of paragraph 79 or 80, or the composition of any one of paragraphs 81 to 88, wherein the tumor has a TMB of at least about 10 mutations per megabase of the sequenced genome. [Section 90] 90. The method or composition of any one of paragraphs 1 to 89, wherein the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer and melanoma. [Section 91] 90. The method or composition of any one of paragraphs 1 to 89, wherein the tumor is derived from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer or melanoma. [Section 92] 92. The method or composition of any one of paragraphs 1 to 91, wherein the tumor is derived from non-small cell lung cancer (NSCLC) or small cell lung cancer. [Section 93] 93. The method or composition of any one of paragraphs 1 to 92, wherein the tumor is derived from NSCLC. [Section 94] 94. The method or composition of any one of paragraphs 1 to 93, wherein the tumor is derived from stage IV NSCLC. [Section 95] 95. The method or composition of paragraph 93 or 94, wherein the NSCLC is squamous NSCLC. [Section 96] 95. The method or composition of paragraph 93 or 94, wherein the NSCLC is non-squamous NSCLC. [Section 97] 97. The method or composition of any one of paragraphs 1 to 96, wherein the tumor is locally advanced, progressive, or metastatic. [Section 98] 98. The method or composition of any one of paragraphs 1 to 97, wherein the tumor is refractory or recurrent. [Section 99] 99. The method or composition of any one of paragraphs 1 to 98, wherein the tumor is refractory after at least one prior therapy for treating the tumor, wherein the at least one prior therapy comprises a standard therapy. [Section 100] 100. The method or composition of paragraph 99, wherein the at least one prior therapy comprises prior chemotherapy. [Section 101] 101. The method or composition of paragraph 100, wherein the prior chemotherapy is platinum-based chemotherapy. [Section 102] 102. The method or composition of any one of paragraphs 1 to 101, wherein at least 1% of the tumor cells exhibit membrane PD-L1 expression. [Section 103] 103. The method or composition of any one of paragraphs 1 to 102, wherein at least 5% of the tumor cells exhibit membrane PD-L1 expression. [Section 104] 104. The method or composition of any one of paragraphs 1 to 103, wherein the anti-PD-1 antibody and the chemotherapeutic agent are administered on the same day. [Section 105] 104. The method or composition of any one of paragraphs 1 to 103, wherein the anti-PD-1 antibody and the chemotherapeutic agent are administered on different days. [Section 106] 105. The method or composition of any one of paragraphs 1 to 104, wherein the dose of anti-PD-1 antibody, the dose of anti-CTLA-4 antibody, and the dose of chemotherapeutic agent are all administered on the same day. [Section 107] (i) the anti-PD-1 antibody is administered once every three weeks; (ii) an anti-CTLA-4 antibody is administered once every six weeks; and (iii) the chemotherapy agent is administered once every three weeks for two cycles; wherein the dose of anti-PD-1 antibody, the dose of anti-CTLA-4 antibody, and the dose of chemotherapy agent are all administered on day 1 of a first 3-week cycle. 105. The method or composition of any one of paragraphs 1 to 104. [Section 108] 108. The method or composition of paragraph 107, wherein the dose of anti-PD-1 antibody and the dose of chemotherapeutic agent are administered on day 1 of a second 3-week cycle. [Section 109] 109. The method or composition of paragraph 107 or 108, wherein the dose of anti-PD-1 antibody and the dose of anti-CTLA-4 antibody are administered on day 1 of a third 3-week cycle. [Section 110] 110. The method or composition of any one of paragraphs 1 to 109, wherein the period between the first administration after the induction phase and the last administration of the induction phase is equal to or less than about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days (2 weeks), about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days (1 month), about 31 days (1 month), about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, or about 3 months. [Section 111] 111. The method or composition of any one of paragraphs 1 to 98 and 102 to 110, wherein the tumor is not refractory to chemotherapy during or after the induction period.

[0009] One aspect of the invention relates to a method of treating a tumor in a subject in need thereof, comprising administering to the subject an anti-PD-1 antibody or an anti-PD-L1 antibody, wherein prior to administering the anti-PD-1 antibody or anti-PD-L1 antibody, the subject undergoes an induction phase comprising chemotherapy for a duration that is shorter than the standard duration of the chemotherapeutic agent.

[0010] In some embodiments, the induction phase further comprises administering an anti-PD-1 antibody or an anti-PD-L1 antibody. In some embodiments, the method further comprises administering an antibody that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody"), or an antigen-binding portion thereof. In some embodiments, the anti-CTLA-4 antibody is administered during the induction phase; in some embodiments, the anti-CTLA-4 antibody is administered after the induction phase. In some embodiments, the anti-CTLA-4 antibody is administered before or after the anti-PD-1 antibody or anti-PD-L1 antibody. In some embodiments, the anti-CTLA-4 antibody is administered after chemotherapy. In some embodiments, the anti-CTLA-4 antibody is administered simultaneously with the anti-PD-1 antibody or anti-PD-L1 antibody.

[0011] In some embodiments, the chemotherapeutic agent is administered for fewer than 10 cycles, fewer than 9 cycles, fewer than 8 cycles, fewer than 7 cycles, fewer than 6 cycles, fewer than 5 cycles, fewer than 4 cycles, or fewer than 3 cycles. In some embodiments, the chemotherapeutic agent is administered for 5 cycles, 4 cycles, 3 cycles, 2 cycles, or 1 cycle. In some embodiments, the chemotherapeutic agent is administered for fewer than 5 cycles. In some embodiments, the chemotherapeutic agent is administered for fewer than 4 cycles. In some embodiments, the chemotherapeutic agent is administered for fewer than 3 cycles. In some embodiments, the chemotherapeutic agent is administered for fewer than 2 cycles. In some embodiments, the chemotherapeutic agent is administered for no more than 2 cycles.

[0012] In some embodiments, the chemotherapy comprises standard therapy. In some embodiments, the chemotherapeutic agent comprises an alkylating agent, an antimetabolite, an anti-microtubule agent, a topoisomerase inhibitor, a cytotoxic antibiotic, or any combination thereof.

[0013] In some embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent. In some embodiments, the chemotherapeutic agent comprises cisplatin, oxaliplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, or any combination thereof. In some embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and a second agent. In some embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and paclitaxel. In some embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and pemetrexed. In some embodiments, the chemotherapeutic agent comprises carboplatin and paclitaxel. In some embodiments, the chemotherapeutic agent comprises carboplatin and pemetrexed. In some embodiments, the chemotherapeutic agent comprises cisplatin and pemetrexed. In some embodiments, the chemotherapeutic agent is a histology-based chemotherapeutic agent.

[0014] In some embodiments, the chemotherapy agent is administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, or about once every 6 weeks. In some embodiments, each cycle is 3 weeks. In some embodiments, the chemotherapy agent is administered on day 1 of each 3-week cycle.

[0015] In some embodiments, chemotherapy consists of carboplatin AUC 6 and paclitaxel 200 mg / m on day 1 of each 3-week cycle. 2 In some embodiments, the chemotherapy comprises administering carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m 2 In some embodiments, the chemotherapy comprises administering cisplatin 75 mg / m 2 and pemetrexed 500 mg / m 2 The method includes administering

[0016] In some embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In some embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In some embodiments, the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. In some embodiments, the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the anti-PD-1 antibody is pembrolizumab.

[0017] In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from 0.1 mg to 10.0 mg per kg of body weight once every two weeks, three weeks, or four weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of 3 mg, 5 mg, or 10 mg per kg of body weight once every three weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose about once every week, about once every two weeks, about once every three weeks, or about once every four weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 360 mg about once every three weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 240 mg about once every two weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every four weeks.

[0018] In some embodiments, the anti-PD-L1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. In some embodiments, the anti-PD-L1 antibody comprises a heavy chain constant region of the human IgG1 isotype. In some embodiments, the anti-PD-L1 antibody cross-competes with an antibody selected from atezolizumab, durvalumab, and avelumab for binding to human PD-L1. In some embodiments, the anti-PD-L1 antibody binds to the same epitope on human PD-L1 as an antibody selected from atezolizumab, durvalumab, and avelumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab, durvalumab, or avelumab.

[0019] In some embodiments, the anti-PD-L1 antibody is administered at a dose range of 0.1 mg to 15.0 mg per kg of body weight once every two weeks, three weeks, or four weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of 3 mg or 5 mg per kg of body weight once every two weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of 10 mg per kg of body weight once every three weeks. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, at least about 1400 mg, or at least about 1500 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, or about every four weeks. In one embodiment, the anti-PD-L1 antibody is administered at a flat dose of about 1200 mg approximately once every three weeks.

[0020] In some embodiments, the anti-CTLA-4 antibody is a chimeric antibody, a humanized antibody, or a human monoclonal antibody or a portion thereof. In some embodiments, the anti-CTLA-4 antibody comprises a heavy chain constant region of a human IgG1 isotype. In some embodiments, the anti-CTLA-4 antibody is ipilimumab. In some embodiments, the anti-CTLA-4 antibody is tremelimumab. In some embodiments, the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

[0021] In some embodiments, the anti-CTLA-4 antibody is administered at a dose range of at least about 0.1 mg to at least about 10.0 mg per kg of body weight about once every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every 6 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 3 mg per kg of body weight about once every 12 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a fixed dose.

[0022] In one embodiment, (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once every three weeks, and (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once every six weeks.

[0023] In some embodiments, (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once every three weeks, (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight once every six weeks, and (iii) the chemotherapeutic agents are carboplatin AUC 6 and paclitaxel 200 mg / m 2 and is administered on day 1 of each 3-week cycle.

[0024] In some embodiments, (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once every three weeks, (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight once every six weeks, and (iii) the chemotherapeutic agents are carboplatin AUC 5 and pemetrexed 500 mg / m 2 and is administered on day 1 of each 3-week cycle.

[0025] In some embodiments, (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once every three weeks, (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight once every six weeks, and (iii) the chemotherapeutic agents are carboplatin AUC 6 and pemetrexed 500 mg / m 2 and is administered on day 1 of an approximately 3-week cycle.

[0026] In one embodiment, (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once every three weeks, (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once every six weeks, and (iii) the chemotherapeutic agent is cisplatin 75 mg / m 2 and pemetrexed 500 mg / m 2 and is administered on day 1 of each 3-week cycle.

[0027] In some embodiments, the chemotherapeutic agent is administered for less than three cycles. In some embodiments, the chemotherapeutic agent is administered for two cycles.

[0028] In certain embodiments, the subject exhibits a progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, subjects exhibit a response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.

[0029] In some embodiments, the tumor has a high tumor mutation burden (TMB) status. In some embodiments, the TMB status is determined by sequencing nucleic acids in the tumor and identifying genomic alterations in the sequenced nucleic acids. In some embodiments, the genomic alterations include: (i) one or more somatic mutations; (ii) one or more non-synonymous mutations; (iii) one or more missense mutations; (iv) one or more alterations selected from the group consisting of base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and combinations thereof; or (v) any combination of (i) to (iv).

[0030] In some embodiments, high TMB is at least 210, at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least In some embodiments, the marker has a score of at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500.

[0031] In some embodiments, the subject's TMB status is compared to a control TMB value, wherein the subject's TMB status is within the highest quantile of the control TMB values, or the subject's TMB status is within the highest tertile of the control TMB values.

[0032] In some embodiments, the biological sample comprises a tumor tissue biopsy, a liquid biopsy, blood, serum, plasma, exoRNA, circulating tumor cells, ctDNA, cfDNA, or any combination thereof.

[0033] In some embodiments, the TMB status is determined by (i) genome sequencing, (ii) exome sequencing, (iii) genomic profiling, or (iv) any combination of (i) through (iii). In some embodiments, the genomic profile is determined by a combination of ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PDCD1LG2, RBM10, STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGFRA, RET, STK11, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTOR, SUFU, AKT1, BRD4, CREBBP, FANCF, GID4(C17o rf39), KAT6A(MYST3), MRE11A, PDK1, RNF43, SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B, ROS1, TAF1, AKT3, BTG1, CRLF2 , FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR, TBX3, ALK, BTK, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER1(FAM123B), C11 orf30(EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1T1, TERT (promoter only), APC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R1 , SDHA, TET2, AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2, SDHB, TGFBR2, ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PL CG2, SDHC, TNFAIP3, ARFRP1, CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF14, ARID1A, CCND2, DAXX, FGF23, GSK3B, KMT2A (MLL), NF1, POLD1, SETD2, TOP1, ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, TOP2A, ARID2, CCNE1, DICER1, FGF4,<h2 style=";text-align:left;direction:ltr">HGF、KMT2D(MLL2)、NFE2L2、PPP2R1A、SLIT2、TP53、ASXL1、CD274、DNMT3A、 FGF6、HNF1A、KRAS、NFKBIA、PRDM1、SMAD2、TSC1、ATM、CD79A、DOT1L、FGFR1 、HRAS、LMO1、NKX2-1、PREX2、SMAD3、TSC2、ATR、CD79B、EGFR、FGFR2、HSD3B 1、LRP1B、NOTCH1、PRKAR1A、SMAD4、TSHR、ATRX、CDC73、EP300、FGFR3、HSP90 AA1、LYN、NOTCH2、PRKCI、SMARCA4、U2AF1、AURKA、CDH1、EPHA3、FGFR4、IDH 1、LZTR1、NOTCH3、PRKDC、SMARCB1、VEGFA、AURKB、CDK12、EPHA5、FH、IDH2、M AGI2、NPM1、PRSS8、SMO、VHL、AXIN1、CDK4、EPHA7、FLCN、IGF1R、MAP2K1、NR AS、PTCH1、SNCAIP、WISP3、AXL、CDK6、EPHB1、FLT1、IGF2、MAP2K2、NSD1、PTE N、SOCS1、WT1、BAP1、CDK8、ERBB2、FLT3、IKBKE、MAP2K4、NTRK1、PTPN11、SO X10, XPO1, BARD1, CDKN1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2、BCL2、CDKN1B、ERBB4、FOXL2、IL7R、MCL1、NTRK3、RAC1、SOX9、ZNF21 7、BCL2L1、CDKN2A、ERG、FOXP1、INHBA、MDM2、NUP93、RAD50、SPEN、ZNF703、B CL2L2、CDKN2B、ERRFI1、FRS2、INPP4B、MDM4、PAK3、RAD51、SPOP、BCL6、CDK N2C、ESR1、FUBP1、IRF2、MED12、PALB2、RAF1、SPTA1、BCOR、CEBPA、EZH2、GAB RA6、IRF4、MEF2B、PARK2、RANBP2、SRC、BCORL1、CHD2、FAM46C、GATA1、IRS2 、MEN1、PAX5、RARA、STAG2、BLM、CHD4、FANCA、GATA2、JAK1、MET、PBRM1、RB1、STAT3, and any combination thereof.

[0034] In some embodiments, (i) the tumor comprises non-small cell carcinoma, (ii) the tumor is relapsed or refractory after at least one prior selective therapy for treating the tumor, or (iii) both (i) and (ii).

[0035] In some embodiments, the genomic profile is obtained from FOUNDATIONONE® CDX (商標) In some embodiments, the tumor has a TMB of at least about 10 mutations per megabase of the sequenced genome.

[0036] In some embodiments, the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer, and melanoma. In some embodiments, the tumor is derived from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer, lung cancer, or melanoma. In some embodiments, the tumor is derived from non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC). In some embodiments, the tumor is derived from NSCLC. In some embodiments, the tumor is derived from stage IV NSCLC. In some embodiments, the NSCLC is squamous NSCLC. In some embodiments, the NSCLC is non-squamous NSCLC.

[0037] In some embodiments, the tumor is locally advanced, progressive, or metastatic. In some embodiments, the tumor is refractory or recurrent. In some embodiments, the tumor is refractory after at least one previous therapy for treating the tumor, wherein at least one previous therapy comprises standard therapy. In some embodiments, the at least one previous therapy comprises a previous chemotherapy. In some embodiments, the previous chemotherapy is a platinum-based chemotherapy.

[0038] In some embodiments, at least 1% of the tumor cells exhibit membrane PD-L1 expression. In some embodiments, at least 5% of the tumor cells exhibit membrane PD-L1 expression.

[0039] In some embodiments, the anti-PD-1 antibody and the chemotherapeutic agent are administered on the same day. In some embodiments, the anti-PD-1 antibody and the chemotherapeutic agent are administered on different days. In some embodiments, the dose of the anti-PD-1 antibody, the dose of the anti-CTLA-4 antibody, and the dose of the chemotherapeutic agent are all administered on the same day.

[0040] In some embodiments, (i) the anti-PD-1 antibody is administered once every 3 weeks, (ii) the anti-CTLA-4 antibody is administered once every 6 weeks, and (iii) the chemotherapy agent is administered once every 3 weeks for two cycles, wherein the dose of the anti-PD-1 antibody, the dose of the anti-CTLA-4 antibody, and the dose of the chemotherapy agent are all administered on day 1 of the first 3-week cycle. In some embodiments, the dose of the anti-PD-1 antibody and the dose of the chemotherapy agent are administered on day 1 of the second 3-week cycle. In some embodiments, the dose of the anti-PD-1 antibody and the dose of the anti-CTLA-4 antibody are administered on day 1 of the third 3-week cycle.

[0041] In certain embodiments, the period between the first administration after the induction phase and the last administration of the induction phase is equal to or less than about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days (2 weeks), about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days (1 month), about 31 days (1 month), about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, or about 3 months.

[0042] In some embodiments, the tumor is not refractory to the chemotherapeutic agent during or after the induction phase.

[0043] Other features and advantages of the present invention will be apparent from the following detailed description and examples, which should not be construed as limiting. The contents of all references cited herein, including scientific literature, press reports, GenBank entries, patents, and patent applications, are expressly incorporated herein by reference. [Brief explanation of the drawings]

[0044] [Figure 1] Figure 1 shows the study design for the safety lead-in study to evaluate safe dose levels of nivolumab and ipilimumab administered with histology-based platinum doublet chemotherapy lead-in treatment. NSCLC = non-small cell lung cancer; SQ = squamous; NSQ = non-squamous; DLT = dose-limiting toxicity. [Figure 2] Figure 2 shows the study design for a clinical trial evaluating the safety and efficacy of treating stage IV NSCLC with a combination of nivolumab and ipilimumab and induction therapy including histology-based platinum-doublet chemotherapy. SQ = squamous; NSQ = non-squamous. DETAILED DESCRIPTION OF THE INVENTION

[0045] Detailed Description of the Invention The present invention provides methods for treating a subject suffering from a tumor, comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) a post-induction phase comprising, after (1), administering to the subject an antibody that specifically binds to PD-1 (an "anti-PD-1 antibody"), or an antigen-binding portion thereof, or an antibody that specifically binds to PD-L1 (an "anti-PD-L1 antibody"), or an antigen-binding portion thereof. The present specification further provides methods for treating a tumor in a subject in need thereof, comprising administering to the subject an anti-PD-1 antibody or an anti-PD-L1 antibody, wherein, prior to administering the anti-PD-1 antibody or anti-PD-L1 antibody, the subject undergoes an induction phase comprising a chemotherapeutic agent for a period shorter than the standard period of chemotherapy. In some embodiments, the tumor is derived from NSCLC.

[0046] term In order that the present description may be more readily understood, certain terms are first defined. Unless otherwise expressly defined herein, as used herein, each of the following terms shall have the meaning set forth below. Additional definitions are set forth throughout the present specification.

[0047] "Administering" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Preferred routes of administration for immunotherapeutics, such as anti-PD-1 antibodies or anti-PD-L1 antibodies, include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the term "parenteral administration" refers to methods of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intrathoracic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, intraarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. Other non-parenteral routes include oral, topical, epithelial or mucosal administration routes, such as intranasal, intravaginal, rectal, sublingual or topical administration. Administration can also be, for example, one or more times, multiple times, and / or one or more times over an extended period of time.

[0048] As used herein, an "adverse event" (AE) is an unfavorable and generally unintended or undesired sign (including an abnormal laboratory finding), symptom, or disease associated with the use of a medical treatment. For example, an adverse event may be associated with immune system activation or proliferation of immune system cells (e.g., T cells) in response to the treatment. A medical treatment may have one or more associated AEs, each of which may have the same or different levels of severity. Reference to a method that can "modify an adverse event" refers to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0049] "Antibody" (Ab) includes, but is not limited to, a glycoprotein immunoglobulin comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds that specifically binds to an antigen, or an antigen-binding portion thereof. Each H chain comprises a heavy chain variable region (herein referred to as V HThe heavy chain constant region comprises three constant domains: C H1 , C H2 and C H3 Each light chain comprises a light chain variable region (referred to herein as V L The light chain constant region contains one constant domain, C L Includes V H and V L The regions are further divided into regions of hypervariability called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). H and V L contains three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0050] Immunoglobulins can be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by the heavy chain constant region genes. The term "antibody" includes, by way of example, both natural and non-natural antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; totally synthetic antibodies; and single-chain antibodies. Non-human antibodies can be humanized by recombinant methods to reduce their immunogenicity in humans. Unless explicitly stated, and unless the context dictates otherwise, the term "antibody" also includes antigen-binding fragments or portions of any of the above immunoglobulins, including monovalent and bivalent fragments or portions, and single-chain antibodies.

[0051] An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1). However, an isolated antibody that specifically binds PD-1 may have cross-reactivity to other antigens, such as PD-1 molecules from different species. Moreover, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0052] The term "monoclonal antibody" (mAb) refers to antibody molecules of single molecular composition, i.e., non-naturally occurring preparations of antibody molecules essentially identical in primary sequence, which display a single binding specificity and affinity for a particular epitope. A monoclonal antibody is an example of an isolated antibody. Monoclonal antibodies may be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0053] A "human antibody" (HuMAb) refers to an antibody having a variable region in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human antibody" and "fully human antibody" are used synonymously.

[0054] "Humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDRs of a non-human antibody have been replaced with corresponding amino acids from a human immunoglobulin. In one embodiment of a humanized form of an antibody, some, most, or all of the amino acids outside the CDRs have been replaced with amino acids from a human immunoglobulin, while some, most, or all of the amino acids within one or more CDRs remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are permissible as long as they do not eliminate the antibody's ability to bind to a particular antigen. A "humanized antibody" retains the same antigen-binding specificity as the original antibody.

[0055] By "chimeric antibody" is meant an antibody whose variable region is derived from one species and whose constant region is derived from another species, e.g., an antibody whose variable region is derived from a murine antibody and whose constant region is derived from a human antibody.

[0056] An "anti-antigen antibody" refers to an antibody that specifically binds to an antigen. For example, an anti-PD-1 antibody specifically binds to PD-1, an anti-PD-L1 antibody specifically binds to PD-L1, and an anti-CTLA-4 antibody specifically binds to CTLA-4.

[0057] An "antigen-binding portion" (also called an "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to the antigen bound by the full-length antibody.

[0058] "Cancer" refers to a broad group of diverse diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and proliferation results in the formation of malignant tumors that can invade adjacent tissues and metastasize to distant parts of the body through the lymphatic system or bloodstream.

[0059] The term "immunotherapy" refers to the treatment of a subject suffering from a disease or at risk of developing or recurring a disease by methods that involve inducing, enhancing, suppressing, or otherwise modifying the immune response. "Treatment" or "treatment" of a subject refers to any intervention or method performed on a subject, or the administration of an active ingredient to a subject, for the purpose of ameliorating, alleviating, ameliorating, inhibiting, delaying, or preventing the onset, progression, development, severity, or recurrence of symptoms, complications, or conditions, or biochemical manifestations associated with a disease.

[0060] "Programmed cell death-1" (PD-1) refers to an immune inhibitory receptor belonging to the CD28 family. PD-1 is expressed primarily on pre-activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one epitope with hPD-1. The complete hPD-1 sequence can be found in GenBank Accession No. U64863.

[0061] "Programmed death ligand-1" (PD-L1) is one of two cell surface glycoprotein ligands for PD-1 (the other is PD-L2) that down-regulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs that share at least one epitope with hPD-L1. The complete hPD-L1 sequence can be found in GenBank Accession No. Q9NZQ7.

[0062] "Cytotoxic T-lymphocyte antigen-4" (CTLA-4) refers to an immune inhibitory receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), its variants, isoforms, and species homologs, as well as analogs that share at least one epitope with hCTLA-4. The complete hCTLA-4 sequence can be found in GenBank Accession No. AAB59385.

[0063] A "subject" includes a human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In a preferred embodiment, the subject is a human. The terms "subject," "patient," and "participant" are used interchangeably herein.

[0064] With respect to the methods and dosages of the present invention, the use of the term "fixed dose" refers to a dose administered to a patient regardless of the patient's weight or body surface area (BSA). Thus, a fixed dose is provided as an absolute amount of drug (e.g., anti-PD-1 antibody) rather than as a mg / kg dose. For example, a 60 kg human and a 100 kg human may receive the same dose of antibody (e.g., 360 mg of anti-PD-1 antibody).

[0065] The use of the term "fixed dose" in reference to the methods of the present invention means that two or more different antibodies (e.g., an anti-PD-1 antibody and an anti-CTLA-4 antibody, or an anti-PD-L1 antibody and an anti-CTLA-4 antibody) in a single composition are present in the composition, particularly in a (fixed) ratio to each other. In some embodiments, the fixed dose is based on the weight (e.g., mg) of the antibody. In certain embodiments, the fixed dose is based on the concentration (e.g., mg / ml) of the antibody. In some embodiments, the ratio is such that the ratio of mg of the first antibody (e.g., anti-PD-1 antibody or anti-PD-L1 antibody) to mg of the second antibody (e.g., anti-CTLA-4 antibody) is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:110, about 1:120, about 1:130, about 1:140, about 1:150, about 1:160, about 1:170, about 1:180, about 1:190, about 1:200, about 1:210, about 1:220, about 1:230, about 1:240, about 1:250, about 1:260, about 1:270, about 1:280, about 1:290, about 1:300, about 1:310, about 1:320, about 1:330, about 1:340, about 1:350, about 1:360, about 1:370, about 1:380, about 1:390, about 1:400, about 1:410, about 1:420, about 1:430, about 1:440, about 1:450, about 1:460, about 1:470, about 1:480, about 1:490, about 1:50 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, or about 2:1. For example, a 3:1 ratio of anti-PD-1 antibody to anti-CTLA-4 antibody may mean that a vial may contain about 240 mg of anti-PD-1 antibody and about 80 mg of anti-CTLA-4 antibody, or about 3 mg / ml of anti-PD-1 antibody and about 1 mg / ml of anti-CTLA-4 antibody.

[0066] As used herein, the term "weight-based dose" means that the dose administered to a patient is calculated based on the patient's weight. For example, if a patient weighing 60 kg requires 3 mg / kg of anti-PD-1 antibody, an appropriate amount of anti-PD-1 antibody can be calculated and used for administration (i.e., 180 mg).

[0067] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is that amount of drug that, when used alone or in combination with another therapeutic agent, protects a subject from developing the disease or promotes disease regression as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods of the disease, or prevention of impairment or disability due to the affliction of the disease. The ability of a therapeutic agent to promote disease regression can be assessed using a variety of methods known to those of skill in the art, such as assaying the activity of a drug in human subjects in clinical trials, animal model systems predictive of efficacy in humans, or in in vitro assays.

[0068] As an example, an "anti-cancer agent" promotes the regression of cancer in a subject. In a preferred embodiment, a therapeutically effective amount of a drug promotes the regression of cancer to the point of eliminating the cancer. "Promoting the regression of cancer" means that the administration of an effective amount of a drug, alone or in combination with an anti-tumor agent, results in a reduction in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free periods, or prevention of functional impairment or disability due to disease affliction. Additionally, the terms "effective" and "effectiveness" in relation to treatment include both pharmacological effectiveness and physiological safety. Pharmacological effectiveness refers to the ability of a drug to promote the regression of cancer in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (side effects) at the cellular, organ, and / or organismal level resulting from the administration of a drug.

[0069] As an example of treating tumors, such as tumors derived from NSCLC, a therapeutically effective amount of an anticancer agent can preferably inhibit cell proliferation or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80%, compared to untreated subjects. In other preferred embodiments of the present invention, tumor regression can be observed, which lasts for at least about 20 days, more preferably at least about 40 days, or even more preferably at least about 60 days. Apart from these final measurements of therapeutic efficacy, the evaluation of immunotherapeutic agents must also take into account immune-related response patterns.

[0070] "Immune response" refers to a biological response in a vertebrate to foreign agents or abnormal, e.g., cancerous, cells, which protects the organism from these agents and the diseases they cause. An immune response is mediated by the action of one or more cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, or neutrophils) and soluble macromolecules (including antibodies, cytokines, and complement) produced by any of these cells or the liver, resulting in the selective targeting, binding, damaging, destroying, and / or eliminating from the vertebrate body an invading pathogen, a pathogen-infected cell or tissue, a cancerous or other abnormal cell, or, in the case of autoimmune or pathological inflammation, a normal human cell or tissue. An immune response can be mediated by, for example, T cells, e.g., effector T cells, Th cells, CD4 + or CD8 + T cells or T reg This includes the activation or inhibition of cells, or any other cells of the immune system, such as NK cells.

[0071] "Immune-related response pattern" refers to a clinical response pattern often observed in cancer patients treated with immunotherapeutic agents that induce cancer-specific immune responses or modify natural immune processes to produce antitumor effects. This response pattern is characterized by a beneficial therapeutic effect observed after an initial increase in tumor burden or the appearance of new lesions, which in conventional chemotherapy evaluations would be classified as disease progression and may be synonymous with drug failure. Therefore, proper evaluation of immunotherapeutic agents may require longitudinal monitoring of the effects of these drugs on the target disease.

[0072] "Immunomodulator" or "immunoregulator" refers to a factor, e.g., a component of a signal transduction pathway, that may be involved in regulating, controlling, or modifying an immune response. "Modulation," "regulation," or "modulation" of an immune response refers to any alteration of cells of the immune system or the activity of such cells (e.g., effector T cells, such as Th1 cells). Such modulation includes stimulation or suppression of the immune system, which may be manifested by an increase or decrease in the number of various cell types, an increase or decrease in the activity of these cells, or any other change that may occur within the immune system. Both inhibitory and stimulatory immunomodulators have been identified, some of which may have enhanced function in the tumor microenvironment. In some embodiments, an immunomodulator targets a molecule on the surface of a T cell. An "immunomodulatory target" or "immunoregulatory target" is a molecule, e.g., a cell surface molecule, that can be targeted for binding by a substance, drug, moiety, compound, or molecule, and whose activity is altered by said binding. Immunomodulatory targets include, for example, cell surface receptors ("immunomodulatory receptors") and receptor ligands ("immunomodulatory ligands").

[0073] "Immunotherapy" refers to the treatment of a subject having or at risk of developing or recurring with a disease by methods that involve inducing, enhancing, suppressing, or otherwise modifying the immune system or immune response. In some embodiments, immunotherapy involves administering an antibody to the subject. In other embodiments, immunotherapy involves administering a small molecule to the subject. In other embodiments, immunotherapy involves administering a cytokine or an analog, variant, or fragment thereof.

[0074] "Immunostimulatory therapy" or "immunostimulatory therapy" refers to a treatment that results in an increased (elicited or enhanced) immune response in a subject, for example, to treat cancer.

[0075] "Enhancing an endogenous immune response" means increasing the efficacy or potency of an existing immune response in a subject. This increase in efficacy and potency can be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response or by stimulating mechanisms that enhance the endogenous host immune response.

[0076] A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is the amount of drug that inhibits the development or recurrence of cancer when administered alone or in combination with an anti-tumor agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or a subject at risk of cancer recurrence. In preferred embodiments, a prophylactically effective amount completely prevents the development or recurrence of cancer. "Inhibiting" the development or recurrence of cancer means either reducing the likelihood of the development or recurrence of cancer, or completely preventing the development or recurrence of cancer.

[0077] As used herein, the term "induction phase" refers to a portion of treatment that prepares a subject for immunotherapy. In certain embodiments, the induction phase is shorter than the post-induction phase. In some embodiments, the induction phase comprises administering a chemotherapeutic agent to the subject. In some embodiments, the induction phase comprises administering a chemotherapeutic agent and an immunotherapeutic agent to the subject, e.g., a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody, or a combination of an anti-PD-L1 antibody and an anti-CTLA-4 antibody. In some embodiments, the induction phase comprises administering a chemotherapeutic agent for a period shorter than the standard period of chemotherapy. In some embodiments, the chemotherapeutic agent is administered for fewer than four cycles. In some embodiments, the chemotherapeutic agent is administered for fewer than three cycles. In some embodiments, the chemotherapeutic agent is administered for fewer than two cycles. In some embodiments, the chemotherapeutic agent is administered over two cycles. In some embodiments, the induction phase comprises administering (i) standard-of-care chemotherapy modified to shorten the duration of chemotherapy; and (ii) immunotherapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody or an anti-PD-L1 antibody and an anti-CTLA-4 antibody. In certain embodiments, the induction phase lasts about 6 weeks. In certain embodiments, the induction phase lasts for two cycles, each cycle being 21 days.

[0078] As used herein, the term "post-induction phase" refers to any period of therapy or treatment that occurs after the induction phase. The post-induction phase can continue for any length of time. In some embodiments, the post-induction phase continues until disease progression, unacceptable adverse events, complete response, or up to two years. In some embodiments, the post-induction phase includes immunotherapy comprising administering an anti-PD-1 antibody and an anti-CTLA-4 antibody, or an anti-PD-L1 antibody and an anti-CTLA-4 antibody. In certain embodiments, the post-induction phase does not include administering a chemotherapeutic agent. In certain embodiments, the post-induction phase begins on day 1 when an immunotherapeutic agent is administered in the absence of a chemotherapeutic agent.

[0079] As used herein, the term "tumor mutational burden" (TMB) refers to the number of somatic mutations in a tumor genome and / or the number of somatic mutations per region of the tumor genome. Germline (inherited) variants are excluded when measuring TMB because the immune system is more likely to recognize them as the tumor's own. Tumor mutational burden (TMB) may also be used interchangeably with "tumor mutational load," "tumor mutational burden," or "tumor mutational load."

[0080] TMB is the genetic analysis of the tumor genome and can therefore be measured using sequencing methods known to those skilled in the art. Tumor DNA can be compared to DNA from matched normal tissue in the patient to exclude germline mutations or polymorphisms.

[0081] In some embodiments, TMB is determined by sequencing tumor DNA using high-throughput sequencing technology, such as next-generation sequencing (NGS) or an NGS-based method. In some embodiments, the NGS-based method is whole genome sequencing (WGS), whole exome sequencing (WES), or FOUNDATIONONE® CDX. (商標)and comprehensive genomic profiling (CGP) of cancer gene panels, such as the MSK-IMPACT clinical trial. In some embodiments, TMB as used herein refers to the number of somatic mutations per megabase (Mb) of sequenced DNA. In one embodiment, TMB is measured using the total number of non-synonymous mutations, such as missense mutations (i.e., mutations of specific amino acids in proteins) and / or nonsense (premature termination of the protein sequence, resulting in truncation), and is identified by normalizing the matched tumor to a germline sample to exclude inherited germline gene mutations. In another embodiment, TMB is measured using the total number of missense mutations in tumors. To measure TMB, a sufficient amount of sample is required. In one embodiment, tissue samples (e.g., at least 10 slides) are used for evaluation. In some embodiments, TMB is expressed as NsMs per megabase (NsMs / Mb). One megabase represents one million bases.

[0082] The TMB status can be a numerical or relative value within the highest quantile or tertile of a control set, eg, a high value, a middle value, or a low value.

[0083] As used herein, the term "high TMB" refers to a number of somatic mutations in a tumor genome that exceeds the normal or average number of somatic mutations. In certain embodiments, the TMB is at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 330, at least 340, at least 350, at least 360, at least 370, at least 380, at least 390, at least 400, at least 410, at least 420, at least 430, at least 440, at least 450, at least 460, at least 470, at least 480, at least 490, at least 500, at least 510, at least 520, at least 530, at least 540, at least 550, at least 555, at least 560, at least 570, at least 580, at least 590, at least 600, at least 610, at least 620, at least 630, at least 640, at least 650, at least 660, at least 670, at least 680, at least 690, at least 700, at least 710, at least 720, at least 730, at least 740, at least 7 at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445 , at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495 or at least 500; in other embodiments, high TMB has a score of at least at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249 or at least 250; and in certain embodiments, high TMB has a score of at least 243.

[0084] In other embodiments, "high TMB" refers to TMB within the highest quantile of control TMB values. For example, all subjects with evaluable TMB data are grouped according to the fractile distribution of TMB, i.e., subjects are ranked from highest to lowest number of genetic mutations and classified into a defined number of groups. In one embodiment, all subjects with evaluable TMB data are ranked and divided into three groups, with "high TMB" being within the highest quantile of control TMB values. In particular embodiments, the tertile boundaries are 0<100 genetic mutations; 100-243 genetic mutations; and >243 genetic mutations. It should be understood that once ranked, subjects with evaluable TMB data can be classified into any number of groups, e.g., quartiles, quintiles, etc.

[0085] In certain embodiments, "high TMB" refers to a TMB of at least about 20 mutations / tumor, at least about 25 mutations / tumor, at least about 30 mutations / tumor, at least about 35 mutations / tumor, at least about 40 mutations / tumor, at least about 45 mutations / tumor, at least about 50 mutations / tumor, at least about 55 mutations / tumor, at least about 60 mutations / tumor, at least about 65 mutations / tumor, at least about 70 mutations / tumor, at least about 75 mutations / tumor, at least about 80 mutations / tumor, at least about 85 mutations / tumor, at least about 90 mutations / tumor, at least about 95 mutations / tumor, or at least about 100 mutations / tumor. In certain embodiments, "high TMB" refers to a TMB of at least about 105 mutations / tumor, at least about 110 mutations / tumor, at least about 115 mutations / tumor, at least about 120 mutations / tumor, at least about 125 mutations / tumor, at least about 130 mutations / tumor, at least about 135 mutations / tumor, at least about 140 mutations / tumor, at least about 145 mutations / tumor, at least about 150 mutations / tumor, at least about 175 mutations / tumor, or at least about 200 mutations / tumor. In particular embodiments, a tumor with high TMB has at least about 100 mutations / tumor.

[0086] "High TMB" can also be used, for example, in mutation assays such as FOUNDATIONONE® CDX (商標) It can refer to the number of mutations per megabase of the sequenced tumor genome as measured by the assay. In one embodiment, high TMB is measured using FOUNDATIONONE® CDX (商標) "High TMB" refers to at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 mutations per megabase of the genome as measured by the assay. In certain embodiments, "high TMB" refers to at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 mutations per megabase of the genome as measured by the assay. (商標) By assay, this means at least 10 mutations per megabase of the genome sequenced.

[0087] As used herein, the term "intermediate TMB" refers to a number of somatic mutations in a tumor genome that is at or about the normal or average number of somatic mutations, and the term "low TMB" refers to a number of somatic mutations in a tumor genome that is less than the normal or average number of somatic mutations. In certain embodiments, a "high TMB" has a score of at least 243, an "intermediate TMB" has a score of 100-242, and a "low TMB" has a score of less than 100 (or 0-100). "Intermediate or low TMB" refers to a tumor genome that is at or about the normal or average number of somatic mutations. (商標) This means fewer than nine mutations per megabase of the sequenced genome, as measured by the assay.

[0088] As used herein, the term "control TMB value" may be the TMB value shown in Table 9.

[0089] In some embodiments, TMB status can be correlated with smoking status. In particular, current or former smokers often have more genetic mutations, e.g., missense mutations, than non-smoking subjects.

[0090] Tumors exhibiting high TMB, such as tumors derived from NSCLC, may also have a high neoantigen load. As used herein, the term "neoantigen" refers to a newly formed antigen not previously recognized by the immune system. A neoantigen can be a protein or peptide recognized by the immune system as foreign (or non-self). Transcription of a gene in a tumor genome carrying a somatic mutation results in mutant mRNA, which, when translated, generates a mutant protein, which is then processed and transported to the ER lumen, where it binds to the MHC class I complex and promotes T cell recognition of the neoantigen. Recognition of neoantigens can promote T cell activation, clonal expansion, and differentiation into effector and memory T cells. Neoantigen load can be correlated with TMB. In some embodiments, TMB is assessed as a surrogate for measuring tumor neoantigen load. The TMB status of a tumor, such as a tumor derived from NSCLC, can be used alone or in combination with other factors as a factor in determining whether a patient is likely to benefit from a particular anti-cancer drug or type of treatment or therapy, such as a combination therapy comprising (a) an anti-PD-1 antibody or anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In one embodiment, a high TMB status (or high TMB) indicates a high likelihood of benefiting from immuno-oncology and can therefore be used to identify patients who are likely to benefit from a combination therapy comprising (a) an anti-PD-1 antibody or anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. Similarly, tumors with high tumor neoantigen load and high TMB are likely to be more immunogenic than tumors with low neoantigen load and low TMB. In addition, high neoantigen / high TMB tumors are likely to be recognized as non-self by the immune system and therefore induce immune-mediated anti-tumor responses. In one embodiment, a high TMB status and a high neoantigen load indicate a high likelihood of benefiting from immuno-oncology, e.g., a combination therapy including: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period that is shorter than the standard period of chemotherapy; and (2) a post-induction phase comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject after (1).As used herein, the term "benefiting from treatment" means improving one or more of overall survival, progression-free survival, partial response rate, complete response rate, and overall response rate, and may include a decrease in tumor growth or size, a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or prevention of disability or incapacity due to the affliction of the disease.

[0091] Other factors, such as environmental factors, may be associated with TMB status. For example, smoking status of NSCLC patients correlated with TMB distribution, such that current and former smokers had higher median TMB values ​​compared with never smokers. See Peters et al., AACR, April 1-5, 2017, Washington, DC. The presence of driver mutations in NSCLC tumors was associated with younger age, female gender, and non-smoker status. See Singal et al., ASCO, June 1-5, 2017; Chicago, IL. A trend was observed linking the presence of driver mutations, such as EGFR, ALK, or KRAS, with lower TMB (P=0.06). See Davis et al., AACR, April 1-5, 2017, Washington, DC.

[0092] As used herein, the term "somatic mutation" refers to an acquired mutation in DNA that occurs after conception. Somatic mutations can occur in any cell in the body except germ cells (sperm and eggs) and therefore are not passed on to offspring. These mutations can, but do not necessarily, cause cancer or other diseases. The term "germline mutation" refers to a genetic change in the body's germ cells (eggs and sperm) that is incorporated into the DNA of every cell in the offspring's body. Germline mutations are passed from parent to offspring. They are also called "hereditary mutations." In TMB analysis, germline mutations are considered a "baseline" and are subtracted from the number of mutations found in a tumor biopsy to determine the TMB in the tumor. Because germline mutations are present in every cell in the body, their presence can be determined by sampling, such as blood or saliva, which is less invasive than a tumor biopsy. Germline mutations increase the risk of developing certain cancers and may play a role in response to chemotherapy.

[0093] The terms "measuring," "measured," or "measurement" refer to determining a measurable amount of somatic mutations in a subject's biological sample, as indicating TMB status. Measurement can be understood to be performed by sequencing nucleic acids in the sample, such as cDNA, mRNA, exoRNA, ctDNA, and cfDNA. Measurements can be performed on the subject's sample and / or control sample(s), and can be, for example, newly detected or correspond to a previous measurement. Measurements can be performed using, for example, PCR, qPCR, Sanger sequencing, genomic profiling (including comprehensive gene panels), exome analysis, genome sequencing, and / or other methods described herein, as known to those skilled in the art. In some embodiments, the measurement identifies genomic alterations in the sequenced nucleic acid. Genomic (or gene) profiling methods can involve a panel of a predetermined set of genes, e.g., 150-500 genes, and in some instances, the genomic alterations assessed in the gene panel correlate with the total somatic mutations assessed. As used herein, the term "gene" when referring to sequencing includes the DNA coding region (e.g., exons), the DNA non-coding regions (e.g., introns and promoters) linked to the coding region, and the mRNA transcript.

[0094] As used herein, the term "genomic alteration" refers to a change (or mutation) in the nucleotide sequence of a tumor's genome (which alteration is not present in the germline nucleotide sequence), and in certain embodiments is a non-synonymous mutation, including, but not limited to, base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and combinations thereof. In certain embodiments, the genomic alteration measured in a biological sample is a missense mutation.

[0095] As used herein, the term "whole genome sequencing" or "WGS" refers to a method of sequencing the entire genome. As used herein, the term "whole exome sequencing" or "WES" refers to a method of sequencing all protein-coding regions (exons) of the genome.

[0096] As used herein, a "cancer genome panel," "hereditary cancer panel," "comprehensive cancer panel," or "multigene cancer panel" refers to a method of sequencing a subset of target cancer genes, including the coding regions, introns, promoters, and / or mRNA transcripts. In some embodiments, a CGP involves sequencing at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, or at least about 50 target cancer genes.

[0097] The terms "genomic profiling assay," "global genomic profiling," or "CGP" refer to an assay that analyzes a panel of genes and selects introns for in vitro diagnosis. CGP combines NGS with targeted bioinformatics analysis to screen for mutations in known cancer genes with clinical relevance. This method can be used to capture mutations missed by testing for "hot spots" (e.g., BRCA1 / BRCA2 mutations or microsatellite markers). In some embodiments, the CGP further includes one or more mRNA transcripts, non-coding RNAs, and / or promoter regions. In one embodiment, the genes in the panel are cancer-associated genes. In another embodiment, the genomic profiling assay is performed using FOUNDATION ONE (登録商標) It is an assay.

[0098] The term "harmonization" refers to a study performed to determine the comparability between two or more measurements and / or diagnostic tests. Harmonization studies provide a systematic approach to address the question of how diagnostic tests compare with each other and with other tests, as well as their interchangeability when used to determine the biomarker status of a patient's tumor. Generally, at least one well-characterized measurement and / or diagnostic test is used as a standard against which other tests are compared. Concordance assessments are often utilized in harmonization studies.

[0099] As used herein, "concordance" refers to the degree of agreement between two measurements and / or diagnostic tests. Concordance can be established using both qualitative and quantitative methods. Quantitative methods for assessing agreement vary depending on the type of measurement. A particular measurement can be expressed as either 1) a categorical / dichotomized variable or 2) a continuous variable. For "categorical / dichotomous variables" (e.g., above or below the TMB cutoff), agreement can be assessed using percentage agreement, such as overall percentage agreement (OPA), positive percentage agreement (PPA), or negative percentage agreement (NPA). For "continuous variables" (e.g., TMB by WES), agreement across the entire spectrum of values ​​can be assessed using Spearman's rank correlation or Pearson's correlation coefficient (r), with values ​​of -1 ≤ r ≤ +1 (note: r = +1 or -1 means that the variables are perfectly correlated). The term "analytical concordance" refers to the degree of agreement in the performance of two assays or diagnostic tests to support clinical use (e.g., identification of biomarkers, genomic alteration types and genomic signatures, and assessment of test reproducibility). The term "clinical concordance" refers to the degree of agreement in how two assays or diagnostic tests correlate with clinical outcomes.

[0100] The term "microsatellite instability" or "MSI" refers to a change in the DNA of certain cells (e.g., tumor cells) in which the number of microsatellites (short repeats of DNA) differs from the number of repeats present in the DNA at inheritance. MSI can be classified as high microsatellite instability (MSI-H) or low microsatellite instability (MSI-L). Microsatellites are short tandem DNA repeats of 1–6 bases. They are prone to DNA replication errors and are repaired by mismatch repair (MMR). Therefore, microsatellites are a good indicator of genomic instability, particularly mismatch repair deficiency (dMMR). MSI is typically diagnosed by screening for five microsatellite markers (BAT-25, BAT-26, NR21, NR24, and NR27). MSI-H indicates the presence of at least two unstable markers out of five analyzed (more than 30% of the markers if a larger panel is used). MSI-L indicates instability of one MSI marker (or 10%-30% of the markers if a larger panel is used). MSS indicates the absence of unstable microsatellite markers.

[0101] As used herein, the term "biological sample" refers to a biological material isolated from a subject. Biological samples can include any biological material suitable for determining TMB, for example, by sequencing nucleic acids in a tumor (or circulating tumor cells) and identifying genomic alterations in the sequenced nucleic acids. Biological samples can be any suitable biological tissue or body fluid, such as tumor tissue, blood, plasma, and serum. In one embodiment, the sample is a tumor tissue biopsy, such as formalin-fixed, paraffin-embedded (FFPE) tumor tissue or fresh-frozen tumor tissue. In another embodiment, the biological sample is a liquid biopsy containing one or more of blood, serum, plasma, circulating tumor cells, exoRNA, ctDNA, and cfDNA.

[0102] As used herein, the terms "about once every week," "about once every two weeks," or other similar dosing interval terms refer to approximate numbers. "About once every week" can include every 7 days ± 1 day, i.e., every 6 to 8 days. "About once every two weeks" can include every 14 days ± 3 days, i.e., every 11 to 17 days. Similar approximations apply, for example, to about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, and about once every 12 weeks. In certain embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose can be administered any day in the first week, followed by the next dose any day in the sixth or twelfth week, respectively. In other embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose is administered on a particular day (e.g., Monday) in week 1, followed by the next dose on the same day (i.e., Monday) in week 6 or week 12, respectively.

[0103] The use of the alternative (e.g., "or") should be understood to mean either one, both, or a combination of the alternatives. As used herein, the indefinite article "a" or "an" should be understood to mean "one or more" of any described or listed component.

[0104] The terms "about" or "essentially comprising" mean a value or composition that is within an acceptable error range of a particular value or composition as determined by one of ordinary skill in the art, which may depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially comprising" can mean within 1 or more than 1 standard deviation, per practice in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 10%. Moreover, particularly with respect to biological systems or processes, these terms can mean up to an order of magnitude or up to 5 times a value. When a particular value or composition is described in this application and claims, unless otherwise specified, the meaning of "about" or "essentially comprising" should be considered to be within an acceptable error range of that particular value or composition.

[0105] Any concentration range, percentage range, ratio range, or integer range described herein should be understood to include all integer values ​​within the recited range, and, where appropriate, fractions thereof (e.g., tenths and hundredths of an integer), unless otherwise stated.

[0106] Various aspects of the invention are described in further detail in the following subsections.

[0107] Methods of the Invention Certain aspects of the present invention relate to methods of treating a subject afflicted with a tumor, comprising administering a combination therapy comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a duration that is shorter than the standard duration of chemotherapy; and (2) a post-induction phase comprising, after (1), administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject. One aspect of the present invention relates to a method of treating a tumor in a subject in need thereof, comprising administering to the subject an anti-PD-1 antibody or an anti-PD-L1 antibody, wherein, prior to administering the anti-PD-1 antibody or anti-PD-L1 antibody, the subject is administered an induction phase comprising a chemotherapeutic agent for a duration that is shorter than the standard duration of chemotherapy. In one embodiment, the tumor is derived from NSCLC.

[0108] Introduction period In certain embodiments, the induction phase comprises administering a chemotherapeutic agent for a period shorter than the standard period of chemotherapy. In some embodiments, the induction phase comprises administering a modified standard therapy, wherein the modified standard therapy is administered for a period shorter than the unmodified standard therapy. In some embodiments, the chemotherapeutic agent is administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, or about once every 6 weeks. In some embodiments, the chemotherapeutic agent is administered for less than 10 cycles, less than 9 cycles, less than 8 cycles, less than 7 cycles, less than 6 cycles, less than 5 cycles, less than 4 cycles, less than 3 cycles, less than 2 cycles, or less than 1 cycle. In some embodiments, the chemotherapeutic agent is administered for 5 cycles, 4 cycles, 3 cycles, 2 cycles, or 1 cycle. In certain embodiments, the chemotherapeutic agent is administered for 2 cycles, wherein each cycle comprises administering the chemotherapeutic agent once about every 3 weeks.

[0109] Standard treatments for different cancer types are well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), an alliance of 21 major cancer centers in the United States, publishes the NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®, the entire contents of which are incorporated herein by reference), which provide detailed, up-to-date information on standard treatments for various cancers (see NCCN Guidelines® (2018) www.nccn.org / professionals / physician_gls / default.aspx (last accessed October 22, 2018)).

[0110] By way of example, the NCCN guidelines for the treatment of NSCLC with chemotherapy include, but are not limited to, treatments selected from the following: (i) for non-squamous subtypes, cisplatin 75 mg / m on day 1 every 21 days for 4 cycles; 2 + pemetrexed 500mg / m on day 1 2 (ii) carboplatin AUC 6 on day 1 and paclitaxel 200 mg / m on day 1 every 21 days for 4 cycles 2 and (iii) for non-squamous disease, carboplatin AUC 5 on day 1 and pemetrexed 500 mg / m on day 1 every 21 days for 4 cycles. 2 See NCCN Guidelines Version 6.2018 Non-Small Cell Lung Cancer. Other standard chemotherapy regimens include (iv) cisplatin 50 mg / m on days 1 and 8 every 28 days for 4 cycles. 2 and vinorelbine 25 mg / m on days 1, 8, 15, 22, and 28. 2 (v) Cisplatin 100 mg / m on day 1 every 28 days for 4 cycles 2 and vinorelbine 30 mg / m on days 1, 8, 15, and 22. 2 (vi) cisplatin 75-80 mg / m on day 1 every 21 days for 4 cycles 2and vinorelbine 25-30 mg / m on days 1 and 8 2 (vii) cisplatin 100 mg / m on day 1 every 21 days for 4 cycles 2 and etoposide 100 mg / m on days 1 to 3 2 (viii) cisplatin 75 mg / m on day 1 every 21 days for 4 cycles 2 and gemcitabine 1250 mg / m on days 1 and 8 2 (ix) Cisplatin 75 mg / m on day 1 every 21 days for 4 cycles 2 and docetaxel 75 mg / m on day 1 2 and (x) carboplatin AUC 5 on day 1 and gemcitabine 1000 mg / m on days 1 and 8 every 21 days for 4 cycles. 2 Thus, in these embodiments where the tumor is derived from NSCLC, for example, stage IV NSCLC, the induction phase comprises administering a chemotherapeutic agent, for example, a modified standard chemotherapeutic agent, for a period shorter than the standard period of chemotherapy, for example, less than 4 cycles. In certain embodiments, the period is less than 3 cycles. In some embodiments, the period is less than 2 cycles. In certain embodiments, the period is 2 cycles. In certain embodiments, the period is at most 2 cycles.

[0111] The chemotherapy administered during the induction phase can include any chemotherapeutic agent or combination known in the art. In certain embodiments, the chemotherapy includes standard therapy, e.g., standard chemotherapy. See NCCN GUIDELINES® (2018). In certain embodiments, the specific chemotherapy administered depends on the histological assessment of the tumor. For example, in some embodiments, the chemotherapeutic agent administered to treat squamous NSCLC differs from the chemotherapeutic agent administered to treat non-squamous NSCLC.

[0112] In some embodiments, the chemotherapeutic agent comprises an alkylating agent, an antimetabolite, an anti-microtubule agent, a topoisomerase inhibitor, a cytotoxic antibiotic, or any combination thereof. In certain embodiments, the chemotherapy comprises a platinum-based chemotherapeutic agent. In some embodiments, the platinum-based chemotherapeutic agent comprises cisplatin, oxaliplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, or any combination thereof.

[0113] In certain embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and a second drug. In some embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and paclitaxel. In other embodiments, the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and pemetrexed. In some embodiments, the chemotherapeutic agent comprises carboplatin and paclitaxel. In some embodiments, the chemotherapeutic agent comprises carboplatin and pemetrexed. In some embodiments, the chemotherapeutic agent comprises cisplatin and pemetrexed. In some embodiments, the chemotherapeutic agent comprises cisplatin and paclitaxel.

[0114] In some embodiments, the induction phase comprises administering chemotherapy, wherein the chemotherapy comprises carboplatin AUC 6 and paclitaxel 200 mg / m on day 1 of each 3-week cycle. 2 In other embodiments, the induction phase comprises administering chemotherapy agents, wherein the chemotherapy agents include carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m on day 1 of each 3-week cycle. 2 In some embodiments, the induction phase comprises administering chemotherapy, wherein the chemotherapy comprises carboplatin AUC 5 and pemetrexed 500 mg / m on day 1 of each 3-week cycle. 2 In some embodiments, the induction phase comprises administering chemotherapy, wherein the chemotherapy comprises carboplatin AUC 6 and pemetrexed 500 mg / m on day 1 of each 3-week cycle. 2In other embodiments, the induction phase comprises administering a chemotherapy agent, wherein the chemotherapy agent comprises cisplatin 75 mg / m on day 1 of each 3-week cycle. 2 and pemetrexed 500 mg / m 2 Includes:

[0115] In some embodiments, the induction phase further comprises administering (i) an anti-PD-1 antibody or anti-PD-L1 antibody, and (ii) an anti-CTLA-4 antibody, in addition to the chemotherapeutic agent. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered on the same day as the chemotherapeutic agent. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered on a different day than the chemotherapeutic agent. In certain embodiments, (i) the anti-PD-1 antibody or anti-PD-L1 antibody and at least one dose of (ii) an anti-CTLA-4 antibody are administered on the same day as the chemotherapeutic agent.

[0116] In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody is administered in a weight-based dose during the induction phase. In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from about 0.1 mg to about 10.0 mg per kg of body weight once every about 2 weeks, about 3 weeks, or about 4 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, or about 10 mg per kg of body weight once every about 2 weeks or about 3 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg per kg of body weight once every about 3 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 3 mg per kg of body weight once every about 3 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 4 mg per kg of body weight once every about 3 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 5 mg / kg of body weight about once every three weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 10 mg / kg of body weight about once every three weeks.

[0117] In some embodiments, the anti-PD-L1 antibody is administered in a weight-based dose during the induction phase. In some embodiments, the anti-PD-L1 antibody is administered at a dose ranging from about 0.1 mg to about 15.0 mg per kg of body weight about once every two weeks, about three weeks, or about four weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 3 mg or about 5 mg per kg of body weight about once every two weeks or about three weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 2 mg per kg of body weight about once every two weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 3 mg per kg of body weight about once every two weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 4 mg per kg of body weight about once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 5 mg per kg of body weight about once every two weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 6 mg / kg body weight once every two weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 7 mg / kg body weight once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 8 mg / kg body weight once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg body weight once every two weeks.

[0118] In some embodiments, the anti-CTLA-4 antibody is administered at a weight-based dose during the induction phase. In some embodiments, the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg to at least about 10.0 mg per kg of body weight about once every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every 6 weeks.

[0119] In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a fixed dose during the induction phase. In certain embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody are both administered at fixed doses. In other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a fixed dose, and the anti-CTLA-4 antibody is administered at a weight-based dose. In yet other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a weight-based dose, and the anti-CTLA-4 antibody is administered at a fixed dose.

[0120] In certain embodiments, the anti-PD-1 antibody is administered at a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 580 mg, at least about 600 mg, at least about 620 mg, at least about 640 mg, at least about 660 mg, at least about 680 mg, at least about 700 mg, or at least about 720 mg during the induction phase. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, or about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 360 mg about once every three weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 240 mg about once every two weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every four weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 720 mg about once every six weeks.

[0121] In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every three weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every four weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every month. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every two months.

[0122] In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, at least about 1400 mg, or at least about 1500 mg during the induction phase. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1200 mg once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1000 mg once every three weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1100 mg once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1500 mg once every three weeks.

[0123] In certain embodiments, the anti-CTLA-4 antibody is administered at a fixed dose of at least about 40 mg, at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, or at least about 500 mg during the induction phase. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, about every four weeks, about every five weeks, about every six weeks, about every seven weeks, about every eight weeks, about every nine weeks, about every ten weeks, about every eleven weeks, or about every twelve weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 80 mg about once every six weeks.

[0124] After the introduction period The post-induction phase begins immediately after the induction phase. In some embodiments, the period between the first administration after the induction phase and the last administration of the induction phase is about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days (2 weeks), about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days (1 month), about 31 days (1 month), about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, or about 3 months, or less. In certain embodiments, the first dose after the induction phase is administered about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days (2 weeks), about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days (1 month), about 31 days (1 month), about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, or less than about 3 months after the last administration during the induction phase. In certain embodiments, the first administration after the induction phase is administered about 3 weeks after the last administration during the induction phase. In certain embodiments, the first dose after the induction phase is administered about 6 weeks after the last dose of the induction phase.

[0125] In certain embodiments, the post-induction phase comprises administering an immunotherapeutic agent without a chemotherapy agent. In some embodiments, the post-induction phase comprises administering an anti-PD-1 antibody or an anti-PD-L1 antibody. In certain embodiments, the post-induction phase further comprises administering an anti-CTLA-4 antibody.

[0126] In some embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered after the induction phase at the same dose as the anti-PD-1 antibody (or anti-PD-L1 antibody) administered during the induction phase. In some embodiments, the anti-CTLA-4 antibody is administered after the induction phase at the same dose as the anti-CTLA-4 antibody administered during the induction phase. In some embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered after the induction phase at a different dose than the anti-PD-1 antibody (or anti-PD-L1 antibody) administered during the induction phase. In some embodiments, the anti-CTLA-4 antibody is administered after the induction phase at a different dose than the anti-CTLA-4 antibody administered during the induction phase.

[0127] In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody is administered in a weight-based dose after the induction period. In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from about 0.1 mg to about 10.0 mg per kg of body weight once every about 2 weeks, about 3 weeks, or about 4 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, or about 10 mg per kg of body weight once every about 2 weeks or about 3 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg per kg of body weight once every about 3 weeks. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 3 mg per kg of body weight once every about 3 weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 4 mg / kg body weight once every three weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 5 mg / kg body weight once every three weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 10 mg / kg body weight once every three weeks.

[0128] In some embodiments, the anti-PD-L1 antibody is administered in a weight-based dose during the post-induction period. In some embodiments, the anti-PD-L1 antibody is administered at a dose ranging from about 0.1 mg to about 15.0 mg per kg of body weight about once every two weeks, about three weeks, or about four weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 3 mg or about 5 mg per kg of body weight about once every two weeks or about three weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 2 mg per kg of body weight about once every two weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 3 mg per kg of body weight about once every two weeks. In a specific embodiment, the anti-PD-L1 antibody is administered at a dose of about 4 mg per kg of body weight about once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 5 mg per kg of body weight about once every two weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 6 mg / kg body weight once every two weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 7 mg / kg body weight once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 8 mg / kg body weight once every two weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg body weight once every two weeks.

[0129] In some embodiments, the anti-CTLA-4 antibody is administered in a weight-based dose after the induction period. In some embodiments, the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg to at least about 10.0 mg per kg of body weight about once every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every 6 weeks.

[0130] In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a fixed dose during the post-induction period. In certain embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody are both administered at fixed doses. In other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a fixed dose, and the anti-CTLA-4 antibody is administered at a weight-based dose. In yet other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a weight-based dose, and the anti-CTLA-4 antibody is administered at a fixed dose.

[0131] In certain embodiments, the anti-PD-1 antibody is administered at a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 580 mg, at least about 600 mg, at least about 620 mg, at least about 640 mg, at least about 660 mg, at least about 680 mg, at least about 700 mg, or at least about 720 mg during the post-induction period. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose about once every week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, or about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 360 mg about once every three weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 240 mg about once every two weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every four weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 720 mg about once every six weeks.

[0132] In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every three weeks after the induction phase. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every four weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every month. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every two months.

[0133] In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, at least about 1400 mg, or at least about 1500 mg during the post-induction period. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1200 mg once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1000 mg once every three weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1100 mg once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1500 mg once every three weeks.

[0134] In certain embodiments, the anti-CTLA-4 antibody is administered at a fixed dose of at least about 40 mg, at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, or at least about 500 mg during the post-induction period. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose about once every week, about every two weeks, about every three weeks, about every four weeks, about every five weeks, about every six weeks, about every seven weeks, about every eight weeks, about every nine weeks, about every ten weeks, about every eleven weeks, or about every twelve weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 80 mg about once every six weeks.

[0135] Certain embodiments described herein relate to a method of treating a subject afflicted with a tumor resulting from stage IV NSCLC, comprising administering to the subject a combination therapy comprising: (1) administering to the subject: (a) carboplatin AUC 6 and paclitaxel 200 mg / m administered on day 1 of each 3-week cycle; 2(b) a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks; and (c) an induction phase comprising administering, per cycle, an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; and (2) a post-induction phase comprising administering to the subject a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks and an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; wherein the post-induction phase is performed after the induction phase. In some embodiments, the induction phase lasts for fewer than four cycles of the chemotherapy. In some embodiments, the induction phase lasts for fewer than three cycles of the chemotherapy. In certain embodiments, the induction phase lasts for fewer than two cycles of the chemotherapy.

[0136] Certain embodiments described herein relate to a method of treating a subject afflicted with a tumor resulting from stage IV NSCLC, comprising administering to the subject a combination therapy comprising: (1) administering to the subject (a) a combination of carboplatin AUC 5 and pemetrexed 500 mg / m administered on day 1 of each 3-week cycle; 2 (b) a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks; and (c) an induction phase comprising administering, per cycle, an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; and (2) a post-induction phase comprising administering to the subject a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks and an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; wherein the post-induction phase is performed after the induction phase. In some embodiments, the induction phase lasts for fewer than four cycles of the chemotherapy. In some embodiments, the induction phase lasts for fewer than three cycles of the chemotherapy. In certain embodiments, the induction phase lasts for fewer than two cycles of the chemotherapy.

[0137] Certain embodiments described herein relate to a method of treating a subject afflicted with a tumor resulting from stage IV NSCLC, comprising administering a combination therapy comprising: (1) administering to the subject: (a) a combination of carboplatin AUC 6 and pemetrexed 500 mg / m administered on day 1 of each 3-week cycle; 2 (b) a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks; and (c) an induction phase comprising administering, per cycle, an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; and (2) a post-induction phase comprising administering to the subject a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks and an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; wherein the post-induction phase is performed after the induction phase. In some embodiments, the induction phase lasts for fewer than four cycles of the chemotherapy. In some embodiments, the induction phase lasts for fewer than three cycles of the chemotherapy. In certain embodiments, the induction phase lasts for fewer than two cycles of the chemotherapy.

[0138] Certain embodiments described herein relate to a method of treating a subject afflicted with a tumor resulting from stage IV NSCLC, comprising administering a combination therapy comprising: (1) administering to the subject: (a) cisplatin 75 mg / m administered on day 1 of each 3-week cycle; 2 and pemetrexed 500 mg / m 2(b) a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks; and (c) an induction phase comprising administering, per cycle, an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; and (2) a post-induction phase comprising administering to the subject a fixed dose of about 360 mg of an anti-PD-1 antibody administered once every three weeks and an anti-CTLA-4 antibody at a dose of about 1 mg per kg of body weight administered once every six weeks; wherein the post-induction phase is performed after the induction phase. In some embodiments, the induction phase lasts for fewer than four cycles of the chemotherapy. In some embodiments, the induction phase lasts for fewer than three cycles of the chemotherapy. In certain embodiments, the induction phase lasts for fewer than two cycles of the chemotherapy.

[0139] In certain embodiments, the subject exhibits a progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, subjects exhibit a response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.

[0140] In one aspect, the methods of the invention, e.g., the combination of nivolumab (e.g., about 360 mg every 3 weeks) plus low dose ipilimumab (e.g., about 1 mg / kg every 6 weeks) and two cycles of chemotherapy (e.g., (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m 2 or (ii) cisplatin 75 mg / m 2 +Pemetrexed 500mg / m 2 ) demonstrates superior overall survival compared with chemotherapy alone for up to four cycles and any subsequent maintenance therapy.

[0141] In one aspect, the methods of the invention, e.g., the combination of nivolumab (e.g., about 360 mg every 3 weeks) plus low dose ipilimumab (e.g., about 1 mg / kg every 6 weeks) and two cycles of chemotherapy (e.g., (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m 2 or (ii) cisplatin 75 mg / m 2 +Pemetrexed 500mg / m 2 ) exhibits a safety profile comparable to that of the known immunotherapy (nivolumab and / or ipilimumab) and chemotherapeutic components.

[0142] Anti-PD-1 antibodies useful in the present invention Anti-PD-1 antibodies known in the art can be used in the compositions and methods described herein. Various human monoclonal antibodies that specifically bind to PD-1 with high affinity are described in U.S. Patent No. 8,008,449. The anti-PD-1 human antibodies described in U.S. Patent No. 8,008,449 have been shown to exhibit one or more of the following characteristics: (a) a 1×10 -antibody activity as determined by surface plasmon resonance using a Biacore biosensor system; -7 M or lower K D (b) binds to human PD-1 in a specific binding domain; (b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) increases T-cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus PD-1; (g) inhibits the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulates an antigen-specific memory response; (i) stimulates an antibody response; and (j) inhibits tumor cell growth in vivo. Anti-PD-1 antibodies that can be used in the present invention include monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, and in certain embodiments, at least five, of the above characteristics.

[0143] Other anti-PD-1 monoclonal antibodies are described in, e.g., U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509, U.S. Patent Publication No. 2016 / 0272708, and PCT Publication Nos. WO2012 / 145493, WO2008 / 156712, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107, WO2015 / 35606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858, WO2016 / 19 7367, WO2017 / 024515, WO2017 / 025051, WO2017 / 123557, WO2016 / 106159, WO2014 / 194302, WO2017 / 040790, WO2017 / 133540, WO2017 / 132827, WO2017 / 024465, WO2017 / 025016, WO2017 / 106061, WO2017 / 19846, WO2017 / 024465, WO2017 / 025016, WO2017 / 132825, and WO2017 / 133540 (each of which is incorporated herein by reference in its entirety).

[0144] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab (OPDIVO®, also known as 5C4, BMS-936558, MDX-1106, and ONO-4538), pembrolizumab (Merck; also known as KEYTRUDA®, lambrolizumab, and MK-3475; see WO2008 / 156712), PDR001 (Novartis; see WO2015 / 112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO2012 / 145493), cemiplimab (Regeneron; also known as REGN-2810; see WO2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), BGB-A317 (tislelizumab; Beigene; see WO2015 / 35606 and US2015 / 0079109), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210; WO2015 / 085847; see Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), TSR-042 (Tesaro Biopharmaceutical; also known as ANB011; see WO2014 / 179664), GLS-010 (Wuxi / Harbin Gloria Pharmaceuticals; also known as WBP3055; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO2014 / 194302), AGEN2034 (Agenus; see WO2017 / 040790), MGA012 (Macrogenics, see WO2017 / 19846), IBI308 (Innovent; see WO2017 / 024465, WO2017 / 025016, WO2017 / 132825 and WO2017 / 133540), and BCD-100 (Biocad).

[0145] In one embodiment, the anti-PD-1 antibody is nivolumab, a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively inhibits interaction with PD-1 ligands (PD-L1 and PD-L2), thus preventing downregulation of anti-tumor T cell function (U.S. Patent No. 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0146] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, for example, in U.S. Patent Nos. 8,354,509 and 8,900,587.

[0147] Anti-PD-1 antibodies that can be used in the compositions and methods described herein also include isolated antibodies that specifically bind to human PD-1 and cross-compete with the anti-PD-1 antibodies described herein, e.g., nivolumab (see, e.g., U.S. Patent Nos. 8,008,449 and 8,779,105; WO 2013 / 173223), for binding to human PD-1. In some embodiments, the anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein (e.g., nivolumab). The ability of antibodies to cross-compete for binding to an antigen indicates that these monoclonal antibodies bind to the same epitope region of the antigen and sterically hinder the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have significantly similar functional properties to the control antibody (e.g., nivolumab) due to their binding to the same epitope region of PD-1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays (e.g., Biacore analysis, ELISA assays, or flow cytometry) (see, e.g., WO2013 / 173223).

[0148] In certain embodiments, the antibody (nivolumab) that cross-competes with human PD-1 for binding or binds to the same epitope region of human PD-1 antibody is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, modified antibodies, or humanized or human antibodies. Such chimeric antibodies, modified antibodies, humanized antibodies, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0149] Anti-PD-1 antibodies that can be used in the compositions and methods described herein also include antigen-binding portions of such antibodies. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0150] Anti-PD-1 antibodies suitable for use in the compositions and methods of the invention are those that bind to PD-1 with high specificity and affinity, inhibit binding of PD-L1 and / or PD-L2, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods described herein, an anti-PD-1 "antibody" includes an antigen-binding portion or fragment that exhibits functional properties similar to those of a full antibody in binding to the PD-1 receptor, inhibiting ligand binding, and upregulating the immune system. In some embodiments, the anti-PD-1 antibody or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1.

[0151] In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from 0.1 mg to 20.0 mg per kg of body weight once every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks, e.g., at a dose ranging from 0.1 mg to 10.0 mg per kg of body weight once every 2 weeks, 3 weeks, or 4 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, or 10 mg per kg of body weight once every 2 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, or 10 mg per kg of body weight once every 3 weeks. In another embodiment, the anti-PD-1 antibody is administered at a dose of about 5 mg / kg body weight once every three weeks. In another embodiment, the anti-PD-1 antibody, e.g., nivolumab, is administered at a dose of about 3 mg / kg body weight once every two weeks. In another embodiment, the anti-PD-1 antibody, e.g., pembrolizumab, is administered at a dose of about 2 mg / kg body weight once every three weeks.

[0152] Anti-PD-1 antibodies useful in the present invention can be administered at a fixed dose of about 100 to about 1000 mg, about 100 to about 900 mg, about 100 to about 800 mg, about 100 to about 700 mg, about 100 to about 600 mg, about 100 to about 500 mg, about 200 to about 1000 mg, about 200 to about 900 mg, about 200 to about 800 mg, about 200 to about 700 mg, about 200 to about 600 mg, about 200 to about 500 mg, about 200 to about 480 mg, or about 240 to about 480 mg. In one embodiment, the anti-PD-1 antibody is administered at a dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 560 mg, at least about 580 mg, at least about 590 mg, at least about 600 mg, at least about 610 mg, at least about 620 mg, at least about 630 mg, at least about 640 mg, at least about 650 mg, at least about 660 mg, at least about 670 mg, at least about 680 mg, at least about 690 mg, at least about 700 mg, at least about 710 mg, at least about 720 mg, at least about 740 mg, at least about 750 mg, at least about 760 mg, at least about 770 mg, at least about 780 mg, at least about 790 mg, at least about 800 mg, at least about 810 mg, at least about 820 mg, at least about 830 mg, at least about 840 mg, at least about 850 mg, at least about 860 mg, at least about 870 mg, at least about 880 mg, at least about 890 mg, at least about 900 mg, at least about 910 mg, at least about 920 mg, at least about 930 mg, at least about 940 mg, at least about 950 mg, at least about 960 mg In another embodiment, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, or about 200 mg to about 500 mg, at an interval of about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.

[0153] In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every three weeks. In other embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every two weeks. In other embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 240 mg about once every two weeks. In other embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 360 mg about once every two weeks. In certain embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every four weeks.

[0154] Anti-PD-L1 antibodies useful in the present invention Because anti-PD-1 antibodies and anti-PD-L1 antibodies target the same signaling pathway and have been shown in clinical trials to exhibit similar levels of efficacy in a variety of cancers, including renal cell carcinoma (see Brahmer et al. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl J Med 366:2443-54; WO2013 / 173223), anti-PD-L1 antibodies can be used in place of anti-PD-1 antibodies in any of the treatment methods described herein. Anti-PD-L1 antibodies known in the art can be used in the compositions and methods of the invention. Examples of anti-PD-L1 antibodies useful in the compositions and methods of the invention include those described in U.S. Patent No. 9,580,507. The anti-PD-L1 human monoclonal antibodies described in U.S. Patent No. 9,580,507 have been shown to exhibit one or more of the following characteristics: (a) a PD-L1 activity of 1x10 as measured by surface plasmon resonance using a Biacore biosensor system; -7 M or lower K D(b) binds to human PD-L1 in a mixed lymphocyte reaction (MLR) assay; (c) increases interferon-γ production in an MLR assay; (d) increases IL-2 secretion in an MLR assay; (e) stimulates antibody responses; and (f) reverses the effects of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies that may be used in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, and in certain embodiments, at least five, of the above characteristics.

[0155] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of BMS-936559 (12A4, also known as MDX-1105; see, e.g., U.S. Pat. No. 7,943,743 and WO 2013 / 173223), atezolizumab (Roche; TECENTRIQ®; MPDL3280A, also known as RG7446; see U.S. Pat. No. 8,217,149; see also Herbst et al. (2013) J Clin Oncol 31(suppl):3000), durvalumab (AstraZeneca; IMFINZ). (商標) , also known as MEDI-4736; see WO2011 / 066389), avelumab (Pfizer; also known as BAVENCIO®, MSB-0010718C; see WO2013 / 079174), STI-1014 (Sorrento; see WO2013 / 181634), CX-072 (Cytomx; see WO2016 / 149201), KN035 (3D Med / Alphamab; see Zhang et al., Cell Discov. 7:3 (March 2017)), LY3300054 (Eli Lilly Co.; see, e.g., WO2017 / 034916), and CK-301 (Checkpoint Therapeutics; Gorelik et al., AACR:Abstract 4606 (Apr 2016).

[0156] In a particular embodiment, the PD-L1 antibody is atezolizumab (TECENTRIQ®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0157] In certain embodiments, the PD-L1 antibody is durvalumab (IMFINZI). (商標) Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0158] In certain embodiments, the PD-L1 antibody is avelumab (BAVENCIO®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0159] Anti-PD-L1 antibodies that may be used in the compositions and methods of the invention also include isolated antibodies that specifically bind to human PD-L1 and cross-compete for binding to human PD-L1 with the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab. In some embodiments, the anti-PD-L1 antibody binds to the same epitope as any of the anti-PD-L1 antibodies described herein (e.g., atezolizumab, durvalumab, and / or avelumab). The ability of antibodies to cross-compete for binding to an antigen indicates that they bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have significantly similar functional properties to the reference antibody (e.g., atezolizumab and / or avelumab) due to their binding to the same epitope region of PD-L1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with atezolizumab and / or avelumab in standard PD-L1 binding assays, such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO2013 / 173223).

[0160] In certain embodiments, antibodies that cross-compete with human PD-L1 antibodies, such as atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1 or bind to the same epitope region of human PD-L1 antibodies are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric, modified, humanized, or human antibodies. Such chimeric, modified, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0161] The anti-PD-L1 antibodies that may be used in the compositions and methods of the invention also include antigen-binding portions of such antibodies. It has been suggested that the antigen-binding function of an antibody may be performed by fragments of a full-length antibody.

[0162] Anti-PD-L1 antibodies suitable for use in the compositions and methods of the invention are those that bind to PD-L1 with high specificity and affinity, inhibit PD-1 binding, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods described herein, an anti-PD-L1 "antibody" includes an antigen-binding portion or fragment that binds to PD-L1 and exhibits functional properties similar to whole antibodies in inhibiting receptor binding and upregulating the immune system. In certain embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1.

[0163] The anti-PD-L1 antibody useful in the present invention can be any PD-L1 antibody that specifically binds to PD-L1, for example, an antibody that cross-competes with durvalumab, avelumab, or atezolizumab for binding to human PD-1, for example, an antibody that binds to the same epitope as durvalumab, avelumab, or atezolizumab. In certain embodiments, the anti-PD-L1 antibody is durvalumab. In other embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab.

[0164] In certain embodiments, the anti-PD-L1 antibody is administered at a dose ranging from about 0.1 mg to about 20.0 mg per kg of body weight once every about 2 weeks, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, or about 20 mg per kg of body weight.

[0165] In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 15 mg / kg body weight about once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg body weight about once every two weeks.

[0166] In other embodiments, the anti-PD-L1 antibodies useful in the present invention are administered at a fixed dose of about 200 to about 1600 mg, about 200 to about 1500 mg, about 200 to about 1400 mg, about 200 to about 1300 mg, about 200 to about 1200 mg, about 200 to about 1100 mg, about 200 to about 1000 mg, about 200 to about 900 mg, about 200 to about 800 mg, about 200 to about 700 mg, about 200 to about 600 mg, about 700 to about 1300 mg, about 800 to about 1200 mg, about 700 to about 900 mg, or about 1100 to about 1300 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least about 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, or at least about 1400 mg, with a dosing interval of about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1000 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1100 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1300 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1400 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1500 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg about once every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 800 mg about once every two weeks.

[0167] Anti-CTLA-4 antibody Anti-CTLA-4 antibodies known in the art can be used in the compositions and methods of the present invention. The anti-CTLA-4 antibodies described herein bind to human CTLA-4 in a manner that disrupts the interaction between CTLA-4 and the human B7 receptor. Because the interaction between CTLA-4 and B7 transmits a signal that leads to the inactivation of T cells bearing the CTLA-4 receptor, disruption of the interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhancing, or prolonging an immune response.

[0168] A human monoclonal antibody that specifically binds to CTLA-4 with high affinity is described in U.S. Patent No. 6,984,720. Other anti-CTLA-4 monoclonal antibodies are described, for example, in U.S. Patent Nos. 5,977,318, 6,051,227, 6,682,736 and 7,034,121 and International Publication Nos. WO2012 / 122444, WO2007 / 113648, WO2016 / 196237 and WO2000 / 037504 (each of which is incorporated herein by reference in its entirety). The anti-CTLA-4 human monoclonal antibody described in U.S. Patent No. 6,984,720 has been shown to exhibit one or more of the following characteristics: (a) a cytoplasmic affinity of at least about 10, as measured by Biacore analysis; 7 M -1 or about 10 9 M -1 or about 10 10 M -1 From 10 11 M -1 or higher equilibrium binding constant (K a (b) specifically binds human CTLA-4 with a binding affinity of at least about 10 3 m -1 s -1 , about 10 4 m -1 s -1 or about 10 5 m -1 s -1 The dynamic binding constant (k a );(c) at least about 10 3 m -1 s -1 , about 104 m -1 s -1 or about 10 5 m -1 s -1 The dynamic dissociation constant (k d and (d) inhibit the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human CTLA-4 and exhibit at least one, at least two, or at least three of the above characteristics.

[0169] In certain embodiments, the CTLA-4 antibody is selected from the group consisting of ipilimumab (also known as YERVOY®, MDX-010, 10D1; see U.S. Pat. No. 6,984,720), MK-1308 (Merck), AGEN-1884 (Agenus Inc.; see WO2016 / 196237), and tremelimumab (AstraZeneca; also known as ticilimumab, CP-675,206; see WO2000 / 037504 and Ribas, Update Cancer Ther. 2(3): 133-39 (2007)). In certain embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0170] In certain embodiments, for use in the compositions and methods described herein, the CTLA-4 antibody is ipilimumab, a fully human, IgG1 monoclonal antibody that inhibits the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma.

[0171] In certain embodiments, the CTLA-4 antibody is tremelimumab.

[0172] In a particular embodiment, the CTLA-4 antibody is MK-1308.

[0173] In a particular embodiment, the CTLA-4 antibody is AGEN-1884.

[0174] Anti-CTLA-4 antibodies that can be used in the compositions and methods of the present invention also include isolated antibodies that specifically bind to human CTLA-4 and cross-compete with any of the anti-CTLA-4 antibodies described herein, such as ipilimumab and / or tremelimumab, for binding to human CTLA-4. In certain embodiments, the anti-CTLA-4 antibody binds to the same epitope as any of the anti-CTLA-4 antibodies described herein (e.g., ipilimumab and / or tremelimumab). The ability of antibodies to cross-compete for binding to an antigen indicates that they bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have significantly similar functional properties to the control antibody (e.g., ipilimumab and / or tremelimumab) due to their binding to the same epitope region of CTLA-4. Cross-competing antibodies can be readily identified based on their ability to cross-compete with ipilimumab and / or tremelimumab in standard CTLA-4 binding assays such as Biacore analysis, ELISA assays or flow cytometry (see, e.g., WO2013 / 173223).

[0175] In certain embodiments, the antibody that cross-competes with human CTLA-4 antibody such as ipilimumab and / or tremelimumab for binding to human CTLA-4 or binds to the same epitope region of human CTLA-4 antibody is a monoclonal antibody.For administration to human subjects, these cross-competing antibodies are chimeric antibodies, modified antibodies, or humanized or human antibodies.Such chimeric antibodies, modified antibodies, humanized antibodies or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0176] Anti-CTLA-4 antibodies that can be used in the compositions and methods of the invention also include antigen-binding portions of such antibodies. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0177] Anti-CTLA-4 antibodies suitable for use in the methods or compositions of the present invention are antibodies that bind to CTLA-4 with high specificity and affinity, inhibit the activity of CTLA-4, and disrupt the interaction of CTLA-4 with the human B7 receptor. In any of the compositions or methods described herein, anti-CTLA-4 "antibodies" include antigen-binding portions or fragments that bind to CTLA-4 and inhibit the interaction of CTLA-4 with the human B7 receptor, and exhibit functional properties similar to those of whole antibodies in upregulating the immune system. In certain embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof cross-competes with ipilimumab and / or tremelimumab for binding to human CTLA-4.

[0178] In certain embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose ranging from about 0.1 mg to about 10.0 mg per kg of body weight once every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In certain embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose of 1 mg or 3 mg per kg of body weight once every 3 weeks, 4 weeks, 5 weeks, or 6 weeks. In one embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose of 3 mg per kg of body weight once every 2 weeks. In another embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose of 1 mg per kg of body weight once every 6 weeks.

[0179] In certain embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a fixed dose of about 10 to about 1000 mg, about 10 to about 900 mg, about 10 to about 800 mg, about 10 to about 700 mg, about 10 to about 600 mg, about 10 to about 500 mg, about 100 to about 1000 mg, about 100 to about 900 mg, about 100 to about 800 mg, about 100 to about 700 mg, about 100 to about 600 mg, about 100 to about 500 mg, about 100 to about 480 mg, or about 240 to about 480 mg. In one embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg , at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 580 mg, at least about 600 mg, at least about 620 mg, at least about 640 mg, at least about 660 mg, at least about 680 mg, at least about 700 mg, or at least about 720 mg. In another embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a fixed dose about once every week, about every two weeks, about every three weeks, about every four weeks, about every five weeks, about every six weeks, about every seven weeks, or about every eight weeks.

[0180] Cytokines In some embodiments, the methods of the present invention comprise administering a combination therapy comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) a post-induction phase comprising administering an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a cytokine to the subject after (1). In some embodiments, the induction phase further comprises administering an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a cytokine.

[0181] The cytokine may be any cytokine known in the art or a variant thereof. In some embodiments, the cytokine is selected from the group consisting of interleukin-2 (IL-2), IL-1β, IL-6, TNF-α, RANTES, monocyte chemoattractant protein (MCP-1), monocyte inflammatory proteins (MIP-1α and MIP-1β), IL-8, lymphotactin, fractalkine, IL-1, IL-4, IL-10, IL-11, IL-13, LIF, interferon-α, TGF-β, and combinations thereof. In some embodiments, the cytokine is a CD122 agonist. In certain embodiments, the cytokine comprises IL-2 or a variant thereof.

[0182] In some embodiments, the cytokine comprises one or more amino acid substitutions, deletions, or insertions compared to the wild-type cytokine amino acid sequence, hi some embodiments, the cytokine comprises an amino acid sequence having at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 amino acid substitutions compared to the amino acid sequence of the wild-type cytokine.

[0183] In some embodiments, the cytokine is modified, e.g., to increase activity and / or half-life. In certain embodiments, the cytokine is modified by fusing a heterologous moiety to the cytokine. The heterologous moiety can be any structure, including a polypeptide, polymer, small molecule, nucleotide, or a fragment or analog thereof. In certain embodiments, the heterologous moiety comprises a polypeptide. In some embodiments, the heterologous moiety comprises albumin or a fragment thereof, albumin-binding polypeptide (ABP), XTEN, Fc, PAS, the C-terminal peptide (CTP) of the beta subunit of human chorionic gonadotropin, or any combination thereof.

[0184] In certain embodiments, the cytokine is modified by fusing the cytokine with a polymer. In some embodiments, the polymer comprises polyethylene glycol (PEG), polypropylene glycol (PPG), hydroxyethyl starch (HES), or any combination thereof. As used herein, "PEG" or "polyethylene glycol" is meant to encompass water-soluble poly(ethylene oxide). Unless otherwise specified, a "PEG polymer" or polyethylene glycol is one in which substantially all (preferably all) monomer subunits are ethylene oxide subunits, although the polymer may include distinct end-capping moieties or functional groups, e.g., for conjugation. PEG polymers for use in the present invention include one of the following two structures: "-(CHCHO)" or "-(CHCHO)" depending on whether the terminal oxygen(s) have been substituted, e.g., during synthetic transformations. n-n " or "-(CH2CH2O) n-1 CH2CH2-". As noted above, for PEG polymers, the variable (n) ranges from about 3 to 4000, and the overall PEG end groups and structure can vary.

[0185] In some embodiments, the methods of the present invention comprise administering combination therapy, including (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) a post-induction phase comprising administering an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a CD122 agonist to the subject after (1). In some embodiments, the induction phase further comprises administering an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a CD122 agonist. In some embodiments, the CD122 agonist comprises IL-2 or a variant thereof. In some embodiments, the CD122 agonist comprises an IL-2 variant having at least one amino acid substitution compared to wild-type IL-2. In some embodiments, the CD122 agonist comprises IL-2 fused to PEG. In some embodiments, the CD122 agonist comprises an IL-2 variant having at least one amino acid substitution compared to wild-type IL-2, wherein the IL-2 variant is fused to PEG.

[0186] Combination therapy In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, and / or anti-CTLA-4 antibody is administered in a therapeutically effective amount. In some embodiments, the methods of the present invention comprise administering therapeutically effective amounts of an anti-PD-1 antibody and an anti-CTLA-4 antibody. In other embodiments, the methods comprise administering therapeutically effective amounts of an anti-PD-L1 antibody and an anti-CTLA-4 antibody. Any anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody described herein can be used in the methods of the present invention. In certain embodiments, the anti-PD-1 antibody comprises nivolumab. In some embodiments, the anti-PD-1 antibody comprises pembrolizumab. In some embodiments, the anti-PD-L1 antibody comprises atezolizumab. In some embodiments, the anti-PD-L1 antibody comprises durvalumab. In some embodiments, the anti-PD-L1 antibody comprises avelumab. In some embodiments, the anti-CTLA-4 antibody comprises ipilimumab. In some embodiments, the anti-CTLA-4 antibody comprises tremelimumab.

[0187] In some embodiments, (a) the anti-PD-1 antibody or anti-PD-L1 antibody and (b) the anti-CTLA-4 antibody are each administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, or about once every 6 weeks. In some embodiments, (a) the anti-PD-1 antibody or anti-PD-L1 antibody and (b) the anti-CTLA-4 antibody are each administered about once every 7 weeks, about once every 8 weeks, about once every 9 weeks, about once every 10 weeks, about once every 11 weeks, or about once every 12 weeks. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered about once every 2 weeks, about once every 3 weeks, or about once every 4 weeks, and the anti-CTLA-4 antibody is administered about once every 6 weeks. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered about once every 2 weeks, about once every 3 weeks, or about once every 4 weeks, and the anti-CTLA-4 antibody is administered about once every 12 weeks. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered on the same day as the anti-CTLA-4 antibody. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered on a different day than the anti-CTLA-4 antibody.

[0188] In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg per kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg per kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 3 mg per kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 4 mg per kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 5 mg / kg body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 10 mg / kg body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of about 6 mg / kg body weight about once every four weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks.

[0189] In certain embodiments, the anti-PD-1 antibody is administered at a flat dose of about 200 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 200 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 240 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 300 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 300 mg about once every four weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 360 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 400 mg about once every four weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 400 mg about once every six weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 400 mg about once every 8 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every 6 weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 480 mg about once every 4 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every 6 weeks. In some embodiments, the anti-PD-1 antibody is administered at a flat dose of about 480 mg about once every 6 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every 6 weeks.

[0190] In certain embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of 240 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 300 mg about once every four weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 360 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every four weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every six weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 400 mg about once every eight weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every four weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 480 mg about once every six weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks.

[0191] In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 2 mg / kg body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 3 mg / kg body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 4 mg / kg body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 5 mg / kg body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 6 mg per kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 7 mg per kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 8 mg per kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 9 mg per kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg of body weight about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks.In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 15 mg / kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of about 15 mg / kg of body weight about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every 12 weeks.

[0192] In certain embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 800 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1000 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1100 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1200 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks. In one embodiment, the anti-PD-L1 antibody is administered at a fixed dose of about 1500 mg once about every three weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight once about every six weeks.

[0193] In certain embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 800 mg about once every two weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1000 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1100 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a fixed dose of about 80 mg about once every six weeks. In some embodiments, the anti-PD-L1 antibody is administered at a flat dose of about 1500 mg about once every three weeks, and the anti-CTLA-4 antibody is administered at a flat dose of about 80 mg about once every six weeks.

[0194] Tumor mutation burden As tumors grow, somatic mutations that are not present in germline DNA accumulate. TMB refers to the number of somatic mutations in the tumor genome (after taking into account germline mutation DNA) and / or the number of somatic mutations per area of ​​the tumor genome. The acquisition of somatic mutations and the resulting higher TMB can be influenced by different mechanisms, such as exposure to exogenous mutagens (e.g., smoking) and DNA mismatch repair mutations (e.g., MSI in colorectal and esophageal cancer). In solid tumors, approximately 95% of mutations are single-base substitutions (Vogelstein et al., Science (2013) 339:1546-1558). "Non-synonymous mutation" herein refers to a nucleotide mutation that changes the amino acid sequence of a protein. Both missense and nonsense mutations can be non-synonymous mutations. "Missense mutation" herein refers to a non-synonymous point mutation in which a single nucleotide change results in a codon that codes for a different amino acid. By "nonsense mutation" herein is meant a nonsynonymous point mutation in which a codon has been changed to a premature stop codon resulting in truncation of the protein produced.

[0195] In one embodiment, somatic mutations can be expressed at the RNA and / or protein level, resulting in neoantigens (also called neoepitopes). Neoantigens can affect immune-mediated anti-tumor responses. For example, recognition of neoantigens can promote T cell activation, clonal proliferation, and differentiation into effector and memory T cells.

[0196] During tumor development, early clonal mutations (or "trunk mutations") may be harbored by most or all tumor cells, whereas later mutations (or "branch mutations") may occur only in a subset of tumor cells or tumor regions (Yap et al., Sci Tranl Med (2012) 4:1-5; Jamai-Hanjani et al., (2015) Clin Cancer Res 21:1258-1266). Consequently, neoantigens derived from clonal "stem" mutations are more widely distributed in the tumor genome than "branch" mutations, potentially resulting in a greater number of T cells reactive against clonal neoantigens (McGranahan et al., (2016) 351:1463-1469). In general, tumors with high TMB also have a higher neoantigen burden, which may result in higher tumor immunogenicity and increased T cell reactivity and antitumor responses. Thus, cancers with high TMB may respond well to immunotherapy, such as treatment with anti-PD-1 or anti-PD-L1 antibodies. See, e.g., International Publication WO / 2018 / 183928 A1, the entire contents of which are incorporated herein by reference.

[0197] Advances in sequencing technology allow for the assessment of tumor genomic mutational status. Sequencing methods known to those skilled in the art can be used to sequence nucleic acids from tumor genomes (e.g., obtained from biological samples from subjects suffering from tumors). In one embodiment, PCR or qPCR, Sanger sequencing, or next-generation sequencing ("NGS") methods (e.g., genomic profiling, exome analysis, or genome sequencing) can be used to measure TMB. In some embodiments, TMB status is measured using genomic profiling. Genomic profiling involves analyzing nucleic acids (including coding and non-coding regions) from tumor samples and can be performed using methods that include optimized nucleic acid selection, read alignment, and mutation calling. In some embodiments, genomic profiling provides next-generation sequencing (NGS)-based analysis of tumors, which can be optimized on a cancer-by-cancer, gene-by-gene, and / or site-by-site basis. Genomic profiling can incorporate the use of multiple individually modified alignment methods or algorithms to optimize performance in sequencing methods, particularly methods that involve massively parallel sequencing of many different genetic events in many different genes. Genomic profiling provides comprehensive analysis of a subject's cancer genome with clinical-grade quality, and the results of genetic analysis can be relevant to relevant scientific and medical knowledge to improve the quality and efficiency of cancer therapy.

[0198] Genomic profiling involves panels of predetermined gene sets including only 5 genes or 1000 genes, about 25 to about 750 genes, about 100 to about 800 genes, about 150 to about 500 genes, about 200 to about 400 genes, about 250 to about 350 genes. In one embodiment, the genomic profile comprises at least 300 genes, at least 305 genes, at least 310 genes, at least 315 genes, at least 320 genes, at least 325 genes, at least 330 genes, at least 335 genes, at least 340 genes, at least 345 genes, at least 350 genes, at least 355 genes, at least 360 genes, at least 365 genes, at least 370 genes, at least 375 genes, at least 380 genes, at least 385 genes, at least 390 genes, at least 395 genes, or at least 400 genes. In another embodiment, the genomic profile comprises at least 325 genes. In particular embodiments, the genomic profile comprises at least 315 cancer-associated genes and introns of 28 genes (FOUNDATIONONE®), or the entire DNA coding sequence of 406 genes, the introns of 31 rearranged genes, and the RNA sequence (cDNA) of 265 genes (FOUNDATIONONE® Heme). In another embodiment, the genomic profile comprises 26 genes and 1000 associated mutations (EXODX® Solid Tumor). In yet another embodiment, the genomic profile comprises 76 genes (Guardant360). In yet another embodiment, the genomic profile comprises 73 genes (Guardant360). In another embodiment, the genomic profile comprises 354 genes and introns of 28 genes for rearrangement (FOUNDATIONONE® CDX). (商標)In a particular embodiment, the genomic profile is FOUNDATIONONE® F1CDx. In another embodiment, the genomic profile is 468 genes (MSK-IMPACT (商標) One or more genes may be added to the genomic profile as additional genes identified as being associated with oncology.

[0199] FOUNDATIONONE® Assay The FOUNDATIONONE® Assay is a comprehensive genomic profiling assay for solid tumors, including, but not limited to, lung, colon, and breast cancer, melanoma, and ovarian cancer. The FOUNDATIONONE® Assay uses hybrid capture, next-generation sequencing to identify genomic alterations (base substitutions, insertions and deletions, copy number changes, and rearrangements) and select genomic features (e.g., TMB and microsatellite instability). The assay covers 322 unique genes, including the entire coding regions of 315 cancer-related genes and selected introns from 28 genes. A complete list of FOUNDATIONONE® Assay genes is shown in Tables 1A and 1B. See FOUNDATIONONE: Technical Specifications, Foundation Medicine, Inc., available at FoundationMedicine.com (incorporated herein by reference in its entirety) (last accessed March 16, 2018).

[0200] [Table 1] [Table 2]

[0201] [Table 3]

[0202] EXODX® Solid Tumor Assay In one embodiment, TMB is measured using the EXODX® Solid Tumor Assay. The EXODX® Solid Tumor Assay is an exoRNA and cfDNA-based assay that detects actionable mutations in cancer pathways. The EXODX® Solid Tumor Assay is a plasma-based assay that does not require tissue samples. The EXODX® Solid Tumor Assay covers 26 genes and 1,000 mutations. The specific genes covered by the EXODX® Solid Tumor Assay are listed in Table 2. See Plasma-Based Solid Tumor Mutation Panel Liquid Biopsy (Exosome Diagnostics, Inc.), available at exosomedx.com (last accessed March 16, 2018).

[0203] [Table 4]

[0204] FOUNDATIONONE® Liquid Assay In one embodiment, TMB is measured using the FOUNDATIONONE® Liquid Assay. The FOUNDATIONONE® Liquid Assay is a cfDNA-based assay that detects circulating tumor DNA (ctDNA). The assay is a plasma-based assay that does not require a solid tumor sample. The FOUNDATIONONE® Liquid Assay covers 70 genes. The specific genes covered by the FOUNDATIONONE® Liquid Assay are listed in Tables 3A-3C. See FOUNDATIONONE® Liqui, Technical Specifications, Foundation Medicine (last accessed October 6, 2018), assets.ctfassets.net / vhribv12lmne / 3SPYAcbGdqAeMsOqMyKUog / d0eb51659e08d733bf39971e85ed940d / F1L_TechnicalInformation_MKT-0061-04.pdf.

[0205] [Table 5]

[0206] [Table 6]

[0207] [Table 7]

[0208] Guardant360 Assay In some embodiments, TMB status is determined using the Guardant360 assay. The Guardant360 assay measures mutations in at least 73 genes (Table 4A), 23 indels (Table 4B), 18 CNVs (Table 4C), and 6 fusion genes (Table 4D). See GuardantHealth.com (last accessed March 16, 2018). In some embodiments, TMB status is determined using the GUARDANTOMNI (商標) Measured using an assay. (商標) The assay is a comprehensive genomic profiling tool that includes a 500-gene panel.

[0209] [Table 8]

[0210] [Table 9]

[0211] [Table 10]

[0212] [Table 11]

[0213] ILLUMINA® TruSight Assays In some embodiments, TMB is determined using the TruSight Tumor 170 Assay (ILLUMINA). The TruSight Tumor 170 Assay is a next-generation sequencing assay that simultaneously analyzes DNA and RNA and covers 170 genes associated with common solid tumors. The TruSight Tumor 170 Assay evaluates fusions, splice variants, insertions / deletions, single nucleotide variants (SNVs), and amplifications. The TruSight Tumor 170 Assay gene list is shown in Tables 5A to 5C.

[0214] [Table 12]

[0215] [Table 13]

[0216] [Table 14]

[0217] FOUNDATIONONE® F1CDx Assay FOUNDATIONONE® CDX (商標) (“F1CDx”) is a next-generation sequencing-based in vitro diagnostic device for the detection of substitution, insertion, and deletion alterations (indels), and copy number alterations (CNAs) in 324 genes and selected gene rearrangements, as well as genomic features (including microsatellite instability (MSI) and tumor mutation burden (TMB)) using DNA isolated from formalin-fixed, paraffin-embedded (FFPE) tumor tissue samples. F1CDx has been approved by the U.S. Food and Drug Administration (FDA) for use in several tumors, including NSCLC, melanoma, breast cancer, colorectal cancer, and ovarian cancer.

[0218] The F1CDx assay uses a single DNA extraction method from conventional FFPE biopsy or surgical resection samples, of which 50–1000 ng is subjected to whole-genome shotgun library construction and hybridization-based capture of all coding exons from 309 cancer-related genes, one promoter region, one non-coding (ncRNA), and selected intronic regions from 34 commonly rearranged genes (21 of which contain coding exons). Tables 6A and 6B provide a complete list of genes included in the F1CDx assay. In total, the assay detects alterations in a total of 324 genes. Using the ILLUMINA® HiSeq4000 platform, hybrid capture-selected libraries are sequenced to high uniform depth (targeting a median coverage of >500X with >100X coverage and >99% of exons). The sequence data are then processed using a customized analysis pipeline designed to detect all classes of genomic alterations, including base substitutions, indels, copy number changes (amplifications and homozygous gene deletions), and selected genomic rearrangements (e.g., gene fusions). Additionally, genomic features including microsatellite instability (MSI) and tumor mutation burden (TMB) have been reported.

[0219] [Table 15] [Table 16]

[0220] [Table 17]

[0221] The F1CDx assay identifies a variety of alterations in gene and / or intron sequences, including substitutions, insertions / deletions, and CNAs. The F1CDx assay was previously identified as concordant with an externally validated NGS assay and the FOUNDATIONONE® (F1 LDT) assay. FOUNDATIONONE® CDX available at FoundationMedicine.com (商標) : Technical Information, Foundation Medicine, Inc. (incorporated herein by reference in its entirety) (last accessed March 16, 2018).

[0222] MSK-IMPACT (商標) In some embodiments, the TMB status is determined by MSK-IMPACT (商標) Assay: MSK-IMPACT (商標) The assay uses next-generation sequencing to analyze the mutation status of 468 genes. The target genes are identified using the ILLUMINA HISEQ (商標) The device captures and sequences the samples. (商標) The assay is US FDA cleared for the detection of somatic mutations and microsatellite instability in solid malignancies. MSK-IMPACT (商標) The complete list of 468 genes analyzed by the assay is shown in Table 7 (see U.S. Food and Drug Administration FDA decision summary, "Evaluation of Automatic Class III Designation for MSK-IMPACT (Integrated Mutation Profiling of Actionable Cancer Targets)," available at accessdata.fda.gov (last accessed November 15, 2017).

[0223] [Table 18] [Table 19] [Table 20]

[0224] NEOGENOMICS(R) NEOTYPE (商標) Assay In some embodiments, the TMB is NEOGENICS® NEOTYOPE (商標) In some embodiments, TMB is determined using a NEOTYPE assay. (商標) The TMB is determined using a discovery profile. In some embodiments, the TMB is determined using the NEOTYPE solid tumor profile. The NEOGENICS assay measures the number of non-synonymous DNA coding sequence changes per megabase of sequenced DNA.

[0225] ONCOMINE (商標) Tumor mutation burden assay In some embodiments, TMB is administered by THERMOFISHER SCIENTIFIC® ONCOMINE (商標) In some embodiments, TMB is determined using a tumor mutation assay. (商標) ONCOMINE (商標) Determined using tumor mutation assays. (商標) ONCOMINE (商標) The tumor mutation assay is a targeted NGS assay that quantifies somatic mutations to determine tumor mutation burden. The assay covers 1.7 Mb of DNA.

[0226] NOVOGENE (商標) NOVOPM (商標) Assay In some embodiments, TMB is (商標) NOVOPM (商標)In some embodiments, TMB is determined using an assay. (商標) NOVOPM (商標) Determined using a cancer panel assay. (商標) NOVOPM (商標) The cancer panel assay analyzes the entire coding region of 548 genes and the introns of 21 genes (representing approximately 1.5 Mb of DNA), and is a comprehensive NGS cancer panel suitable for the diagnosis and / or treatment of solid tumors according to the National Comprehensive Cancer Network (NCCN) guidelines and medical literature. The assay detects genomic abnormalities such as SNVs, indels, fusions, and copy number variations (CNVs).

[0227] Other TMB assays In some embodiments, TMB is determined using a TMB assay from CARIS® Life Sciences. In some embodiments, TMB is determined using the PESONALIS® ACE ImmunoID assay. In some embodiments, TMB is determined using the PGDX® CANCERXOME assay. (商標) - Determined using the R assay.

[0228] In yet another particular embodiment, genomic profiling detects all variants, i.e., single nucleotide variants, insertions / deletions (indels), copy number variations and rearrangements, e.g., translocations, expression and epigenetic markers.

[0229] A comprehensive gene panel often includes predetermined genes selected based on the tumor type being analyzed. Thus, the genomic profile used to measure TMB status can be selected based on the tumor type that the subject suffers from. In one embodiment, the genomic profile can include a gene set specific to solid tumors. In another embodiment, the genomic profile can include a gene set specific to hematological malignancies and sarcomas.

[0230] Examples include ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PDCD1 LG2, RBM10, STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGFRA RET, STK11, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTOR. SUFU, AKT1, BRD4, CREBBP, FANCF, GID4(C17orf39), KAT6A(MYST3), MRE11A. PDK1, RNF43, SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B, R.S OS1, TAF1, AKT3, BTG1, CRLF2, FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR, TB X3, ALK, BTK, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER1( FAM123B) C11orf30(EMSY) CTCF FAT1 GNAQ KDR MUTYH PIK3CG RUNX1T1 TERT, APC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R1. SDHA, TET2, AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2, S.K DHB, TGFBR2, ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PLCG2, SDHC, and TNFA IP3, ARFRP1, CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF14 ARID1A, CCND2, DAXX, FGF23, GSK3B, KMT2A(MLL), NF1, POLD1, SETD2, TOP1 ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, TOP2A. ARID2, CCNE1, DICER1, FGF4, HGF, KMT2D(MLL2), NFE2L2, PPP2R1A, SLIT2, T P53, ASXL1, CD274, DNMT3A, FGF6, HNF1A, KRAS, NFKBIA, PRDM1, SMAD2, TSC1.ATM, CD79A, DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR , ATRX, CDC73, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMAR CB1, VEGFA, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, W ISP3, AXL, CDK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BARD1, CDKN1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BCL2L1, CDKN2A, ERG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BC In other embodiments, TMB analysis includes one or more genes selected from the group consisting of L6, CDKN2C, ESR1, FUBP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and combinations thereof.It further includes identifying a genomic modification in one or more of ETV6 and MYB.

[0231] These are ABL1, 12B, ABL2, ACTB, ACVR1, and ACVR1 B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, AMER1( FAM123B, AMER1(FAM123B), ANKRD11, APC, APH1A, AR, ARAF, ARFRP1, ARHGAP26(GRAF), ARID1A, ARID1B, ARID2, ARID 5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AURKB. AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL10, BC L11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1, BR IP1(BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf30(EMSY), C11orf30, C11orf30(EMSY), CAD, CALR, CARD11, CARM1, CASP 8. CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274(PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CDC4 2. CDC73, CDH1, CDK12, CDK4, CDK6, CDK8, CDKN1A, CDKN1B, CDKN2A, CDKN2Ap14ARF, CDKN2Ap16INK4A, CDKN2B, CDKN2C, CEBPA CENPA, CHD2, CHD4, CHEK1, CHEK2, CIC, CIITA, CKS1B, CPS1, CREBBP, CRKL, CRLF2, CSDE1, CSF1R, CSF3R, CTCF, CTLA-4, CTNN B1, CTNNA1, CTNNB1, CUL3, CUL4A, CUX1, CXCR4, CYLD, CYP17A1, CYSLTR2, DAXX, DCUN1D1, DDR1, DDR2, DDX3X, DH 2, DICER1, DIS3, DNAJB1, DNM2, DNMT1, DNMT3A, DNMT3B, DOT1L, DROSHA, DTX1, DUSP2, DUSP4, DUSP9, E2F3, EBF1.ECT2L, EED, EGFL7, EGFR, EIF1AX, EIF4A2, EIF4E, ELF3, ELP2, EML4, EML4-A LK, EP300, EPAS1, EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2, ERBB3 ERBB4, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, ERF, ERG, ERRFI1, ERRFl1, ESR 1, ETS1, ETV1, ETV4, ETV5, ETV6, EWSR1, EXOSC6, EZH1, EZH2, FAF1, FAM175A FAM46C, FAM58A, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL FAS, FAS(TNFRSF6), FAT1, FBXO11, FBXO31, FBXW7, FGF1, FGF10, FGF12, FG F14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGFR1 FGFR2, FGFR3, FGFR4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH1, FOXA 1. FOXL2, FOXO1, FOXO3, FOXP1, FRS2, FUBP1, FYN, GABRA6, GADD45B, GATA1 GATA2、GATA3、GATA4、GATA6、GEN1、GID4(C17orf39)、GID4(C17orf39)、GLI 1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRIN2A, GR M3, GSK3B, GTSE1, H3F3A, H3F3B, H3F3C, HDAC1, HDAC4, HDAC7, Hedgehog, HER -2 / NEU;ERBB2、HGF、HIST1H1C、HIST1H1D、HIST1H1E、HIST1H2AC、HIST1H2A G, HIST1H2AL, HIST1H2AM, HIST1H2BC, HIST1H2BD, HIST1H2BJ, HIST1H2BK, H IST1H2BO、HIST1H3A、HIST1H3B、HIST1H3C、HIST1H3D、HIST1H3E、HIST1H3F HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HIST2H3C, HIST2H3D, HIST3H3<h2 style=";text-align:left;direction:ltr">HLA-A、HLA-B、HNF1A、HOXB13、HRAS、HSD3B1、HSP90AA1、ICK、ICOSLG、ID3、IDH1、IDH2、IFNGR1、IGF1、IGF1R、IGF2、IKBKE、IKZF1、IKZF2、IKZF3、IL10、IL7R、INHA、INHBA、INPP4A、INPP4B、INPP5D(SHIP)、INPPL1、INSR、IRF1、IRF2、IRF4、I RF8、IRS1、IRS2、JAK1、JAK2、JAK3、JARID2、JUN、K14、KAT6A(MYST3)、KAT6A(MYST3)、KDM2B、KDM4C、KDM5A、KDM5C、KDM6A、KDR、KEAP1、KEL、KIF5B、KIT、KLF4、KLHL6、KMT2A、KMT2A(MLL)、KMT2B、KMT2C、KMT2C(MLL3)、KMT2D、KMT2D(MLL2)、 KNSTRN、KRAS、LAMP1、LATS1、LATS2、LEF1、LMO1、LRP1B、LRRK2、LTK、LYN、LZTR1、MAF、MAFB、MAGED1、MAGI2、MALT1、MAP 2K1、MAP2K1(MEK1)、MAP2K2、MAP2K2(MEK2)、MAP2K4、MAP3、MAP3K1、MAP3K13、MAP3K14、MAP3K6、MAP3K7、MAPK1、MAPK3 、MAPKAP1、MAX、MCL1、MDC1、MDM2、MDM4、MED12、MEF2B、MEF2C、MEK1、MEN1、MERTK、MET、MGA、MIB1、MITF、MKI67、MKNK1、 MLH1、MLLT3、MPL、MRE11A、MRE11A、MSH2、MSH3、MSH6、MSI1、MSI2、MST1、MST1R、MTAP、MTOR、MUTYH、MYC、MYCL” L1)、MYCL(MYCL1)、MYCL1、MYCN、MYD88、MYO18A、MYOD1、NBN、NCOA3 、NCOR1、NCOR2、NCSTN、NEGR1、NF1、NF2、NFE2L2、NFKBIA、NKX2-1、NK NS D1、NT5C2、NTHL1、NTRK1、NTRK2、NTRK3、NUF2、NUP93、NUP98、P2RY8、PAG1、PAK1、PAK3、PAK7、PALB2、PARK2、PARP1、PARP2、PARP3、PASK、PAX3、PAX5、PAX7、PBRM1、PC、PCBP1、PCLO、PDCD1、PDCD1(PD-1)、PDCD11、PDCD1LG2、PDCD1LG2(PD-L2)、PDGFRA、PDGFRB、PDK1、PDPK1、PGR、PHF6、PHOX2B、PIK3C2B、PIK3C2G、PIK3C3、PIK3CA、PIK3CB、PIK3CD、PIK3CG、PIK3R1、PIK3R2、PIK3R3、PIM1、PLCG2、PLK2、PMAIP1、PMS1、PMS2、PNRC1、POLD1、POLE、POT1、PPARG、PPM1D、PPP2、PPP2R1A、PPP2R2A、PPP4R2、PPP6C、PRDM1、PRDM14、PREX2、PRKAR1A、PRKCI、PRKD1、PRKDC、PRSS8、PTCH1、PTEN、PTP4A1、PTPN11、PTPN2、PTPN6(SHP-1)、PTPRD、PTPRO、PTPRS、PTPRT、QKI、R1A、RAB35、RAC1、RAC2、RAD21、RAD50、RAD51、RAD51B、RAD51C、RAD51D、RAD52、RAD54L、RAF1、RANBP2、RARA、RASA1、RASGEF1A、RB1、RBM10、RECQL、RECQL4、REL、RELN、RET、RFWD2、RHEB、RHOA、RICTOR、RIT1、RNF43、ROS1、RPS6KA4、RPS6KB1、RPS6KB2、RPTOR、RRAGC、RRAS、RRAS2、RTEL1、RUNX1、RUNX1T1、RXRA、RYBP、S1PR2、SDHA、SDHAF2、SDHB、SDHC、SDHD、SERP2、SESN1、SESN2、SESN3、SETBP1、SETD2、SETD8、SF3B1、SGK1、SH2B3、SH2D1A、SHOC2、SHQ1、SLIT2、SLX4、SMAD2、SMAD3、SMAD4、SMARCA1、SMARCA4、SMARCB1、SMARCD1、SMC1A、SMC3、SMO、SMYD3、SNCAIP、SOCS1、SOCS2、SOCS3、SOS1、SOX10、SOX17、SOX2、SOX9、SPEN、SPOP、SPRED1、SPTA1、SRC, SRSF2, STAG2, STAT3, STAT4, STAT5A, STAT5B, STAT6, STK11, STK19, STK40, SUFU, SUZ12, SYK, TAF1, TAP1, TAP2, TBL1XR1, TBX3, TCEB1, TCF3, TCF3(E2A), TCF7L2, TCL1A(TCL1 ), TEK, TERC, TERT, TERT promoter, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL2, TMEM127, TMEM30A, TMPRSS2, TMSB4XP8(TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14, TNFRSF17, TOP 1, TOP2A, TP53, TP53BP1, TP63, TRAF2, TRAF3, TRAF5, TRAF7, TSC1, TSC2, TSHR, TUSC3, TYK2, TYRO3, U2AF1, U2AF2, UPF1, VEGFA, VHL, VTCN1, WDR90, WHSC1, WHSC1 (MMSET or NSD2), WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and combinations thereof.

[0232] These are ABL1, 12B, ABL2, ACTB, and ACVR1. ACVR1B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, A MER1(FAM123BまたはWTX), AMER1(FAM123B), ANKRD11, APC, APH1A, AR, ARAF, ARFRP1, ARHGAP26(GRAF), ARID1A, ARID1B, ARID2. ARID5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AU RKB, AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL1 0, BCL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1 BRIP1(BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf30(EMSY), C11orf30, C11orf30(EMSY), CAD, CALR, CARD11, CARM1, CA SP8, CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274(PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CDC 42 CDC73, CDH1, CDK12, CDK4, CDK6, CDK8, CDKN1A, CDKN1B, CDKN2A, CDKN2Ap14ARF, CDKN2Ap16INK4A, CDKN2B, CDKN2C, CEBP A. CENPA, CHD2, CHD4, CHEK1, CHEK2, CIC, CIITA, CKS1B, CPS1, CREBBP, CRKL, CRLF2, CSDE1, CSF1R, CSF3R, CTCF, CTLA-4, CTNN B1, CTNNA1, CTNNB1, CUL3, CUL4A, CUX1, CXCR4, CYLD, CYP17A1, CYSLTR2, DAXX, DCUN1D1, DDR1, DDR2, DDX3X. DH2, DICER1, DIS3, DNAJB1, DNM2, DNMT1, DNMT3A, DNMT3B, DOT1L, DROSHA, DTX1, DUSP2, DUSP4, DUSP9, E2F3.EBF1, ECT2L, EED, EGFL7, EGFR, EIF1AX, EIF4A2, EIF4E, ELF3, ELP2, EML4, E ML4-ALK, EP300, EPAS1, EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2 ERBB3, ERBB4, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, ERF, ERG, ERRFI1, ERRFl 1, ESR1, ETS1, ETV1, ETV4, ETV5, ETV6, EWSR1, EXOSC6, EZH1, EZH2, FAF1, FA M175A, FAM46C, FAM58A, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI FANCL, FAS, FAS(TNFRSF6), FAT1, FBXO11, FBXO31, FBXW7, FGF1, FGF10, FG F12, FGF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9 FGFR1, FGFR2, FGFR3, FGFR4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH 1, FOXA1, FOXL2, FOXO1, FOXO3, FOXP1, FRS2, FUBP1, FYN, GABRA6, GADD45B. STEP1、STEP2、STEP3、STEP4、STEP6、GEN1、GID4(C17orf39)、GID4(C17orf3 9) GLI1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRI N2A, GRM3, GSK3B, GTSE1, H3F3A, H3F3B, H3F3C, HDAC1, HDAC4, HDAC7, Hedge hog、HER-2 / NEU;ERBB2、HGF、HIST1H1C、HIST1H1D、HIST1H1E、HIST1H2AC、H IST1H2AG, HIST1H2AL, HIST1H2AM, HIST1H2BC, HIST1H2BD, HIST1H2BJ, HIS T1H2BK, HIST1H2BO, HIST1H3A, HIST1H3B, HIST1H3C, HIST1H3D, HIST1H3E HIST1H3F, HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HIST2H3C, HIST2H3D<h2 style=";text-align:left;direction:ltr">HIST3H3、HLA-A、HLA-B、HNF1A、HOXB13、HRAS、HSD3B1、HSP90AA1、ICK、ICOSLG、ID3、IDH1、IDH2、IFNGR1、IGF1、IGF1R、IGF2、IKBKE、IKZF1、IKZF2、IKZF3、IL10、IL7R、INHA、INHBA、INPP4A、INPP4B、INPP5D(SHIP)、INPPL1、INSR、IRF1、IRF2、 IRF4、IRF8、IRS1、IRS2、JAK1、JAK2、JAK3、JARID2、JUN、K14、KAT6A(MYST3)、KAT6A(MYST3)、KDM2B、KDM4C、KDM5A、KDM5C、KDM6A、KDR、KEAP1、KEL、KIF5B、KIT、KLF4、KLHL6、KMT2A、KMT2A(MLL)、KMT2B、KMT2C、KMT2C(MLL3)、KMT2D、KMT2D(ML L2)、KNSTRN、KRAS、LAMP1、LATS1、LATS2、LEF1、LMO1、LRP1B、LRRK2、LTK、LYN、LZTR1、MAF、MAFB、MAGED1、MAGI2、MALT1、MAP2K1、MAP2K1(MEK1)、MAP2K2、MAP2K2(MEK2)、MAP2K4、MAP3、MAP3K1、MAP3K13、MAP3K14、MAP3K6、MAP3K7、MAPK1、MAP K3、MAPKAP1、MAX、MCL1、MDC1、MDM2、MDM4、MED12、MEF2B、MEF2C、MEK1、MEN1、MERTK、MET、MGA、MIB1、MITF、MKI67、MKNK1 MYCL(MYC) L1)、MYCL(MYCL1)、MYCL1、MYCN、MYD88、MYO18A、MYOD1、NBN、NCOA3、NCOR1、NCOR2、NCSTN、NEGR1、NF1、NF2、NFE2L2、NFKBIA、NKX 2-1、NKX3-1、NOD1、NOTCH1、NOTCH2、NOTCH3、NOTCH4、NPM1、NRAS、NRG1、NSD1、NT5C2、NTHL1、NTRK1、NTRK2、NTRK3、NUF2、NUP93、NUP98、P2RY8、PAG1、PAK1、PAK3、PAK7、PALB2、PARK2、PARP1、PARP2、PARP3、PASK、PAX3、PAX5、PAX7、PBRM1、PC、PCBP1、PCLO、PDCD1、PDCD1(PD-1)、PDCD11、PDCD1LG2、PDCD1LG2(PD-L2)、PDGFRA、PDGFRB、PDK1、PDPK1、PGR、PHF6、PHOX2B、PIK3C2B、PIK3C2G、PIK3C3、PIK3CA、PIK3CB、PIK3CD、PIK3CG、PIK3R1、PIK3R2、PIK3R3、PIM1、PLCG2、PLK2、PMAIP1、PMS1、PMS2、PNRC1、POLD1、POLE、POT1、PPARG、PPM1D、PPP2、PPP2R1A、PPP2R2A、PPP4R2、PPP6C、PRDM1、PRDM14、PREX2、PRKAR1A、PRKCI、PRKD1、PRKDC、PRSS8、PTCH1、PTEN、PTP4A1、PTPN11、PTPN2、PTPN6(SHP-1)、PTPRD、PTPRO、PTPRS、PTPRT、QKI、R1A、RAB35、RAC1、RAC2、RAD21、RAD50、RAD51、RAD51B、RAD51C、RAD51D、RAD52、RAD54L、RAF1、RANBP2、RARA、RASA1、RASGEF1A、RB1、RBM10、RECQL、RECQL4、REL、RELN、RET、RFWD2、RHEB、RHOA、RICTOR、RIT1、RNF43、ROS1、RPS6KA4、RPS6KB1、RPS6KB2、RPTOR、RRAGC、RRAS、RRAS2、RTEL1、RUNX1、RUNX1T1、RXRA、RYBP、S1PR2、SDHA、SDHAF2、SDHB、SDHC、SDHD、SERP2、SESN1、SESN2、SESN3、SETBP1、SETD2、SETD8、SF3B1、SGK1、SH2B3、SH2D1A、SHOC2、SHQ1、SLIT2、SLX4、SMAD2、SMAD3、SMAD4、SMARCA1、SMARCA4、SMARCB1、SMARCD1、SMC1A、SMC3、SMO、SMYD3、SNCAIP、SOCS1、SOCS2、SOCS3、SOS1、SOX10、SOX17、SOX2、SOX9、SPEN、SPOP、SPRED1, SPTA1, SRC, SRSF2, STAG2, STAT3, STAT4, STAT5A, STAT5B, STAT6, STK11, STK19, STK40, SUFU, SUZ12, SYK, TAF1, TAP1, TAP2, TBL1 XR1, TBX3, TCEB1, TCF3, TCF3(E2A), TCF7L2, TCL1A(TCL1), TEK, TERC, TERT, TERT promoter, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL2 , TMEM127, TMEM30A, TMPRSS2, TMSB4XP8 (TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14, TNFRSF17, TOP1, TOP2A, TP53, TP53BP1, TP63, TRAF2, TRAF3, TRAF5, TRAF7, TSC1, TSC2, TSHR, TUSC3, TYK2, TYRO3, U2AF1, U2AF2, UPF1, VEGFA, VHL, VTCN1, WDR90, WHSC1, WHSC1 (MMSET or NSD2), At least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, or at least about 100 amino acids selected from the group consisting of WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and combinations thereof , at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, at least about 200, at least about 210, at least about 220, at least about 230, at least about 240, at least about 250, at least about 260, at least about 270, at least about 280, at least about 290, or at least about 300 genes.

[0233] In another embodiment, the genomic profile comprises one or more genes selected from the genes listed in Tables 2-14.

[0234] In one embodiment, the TMB status based on genomic profiling is highly correlated with the TMB status based on whole exome sequencing or whole genome sequencing.Evidence shows that the use of genomic profiling assays such as F1CDx assay is consistent with whole exome sequencing assays and / or whole genome sequencing assays.These data support the use of genomic profiling assays as a more effective method for measuring TMB status without compromising the quality of TMB status prediction.

[0235] TMB can be measured using tissue biopsy samples or circulating tumor DNA (ctDNA), cfDNA (cell-free DNA), and / or liquid biopsy samples. ctDNA can be used to measure TMB status following whole-exome or whole-genome sequencing or genomic profiling using available methods, such as those from GRAIL.

[0236] In some embodiments, a subject is identified as suitable for the combination therapy described herein based on measuring TMB status and identifying high TMB. In some embodiments, the TMB score is calculated as the total number of nonsynonymous missense mutations in the tumor as measured by whole exome sequencing or whole genome sequencing. In one embodiment, high TMB is defined as a mutation count of at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 360, at least 370, at least 380, at least 390, at least 400, at least 410, at least 420, at least 430, at least 440, at least 450, at least 460, at least 470, at least 480, at least 490, at least 500, at least 510, at least 520, at least 530, at least 540, at least 550, at least 560, at least 570, at least 580, at least 590, at least 600, at least 610, at least 620, at least 630, at least 640, at least 650, at least 660, at least 670, at least In some embodiments, the marker has a score of at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500.In another embodiment, high TMB has a score of at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250. In certain embodiments, high TMB has a score of at least 243. In other embodiments, high TMB has a score of at least 244. In certain embodiments, high TMB has a score of at least 245. In other embodiments, high TMB has a score of at least 246. In other embodiments, a high TMB has a score of at least 247. In other embodiments, a high TMB has a score of at least 248. In other embodiments, a high TMB has a score of at least 249. In other embodiments, a high TMB has a score of at least 250. In other embodiments, a high TMB has an integer score of 200-300 or higher. In other embodiments, a high TMB has an integer score of 210-290 or higher. In other embodiments, a high TMB has an integer score of 220-280 or higher. In other embodiments, a high TMB has an integer score of 230-270 or higher. In other embodiments, a high TMB has an integer score of 235-265 or higher.

[0237] Alternatively, high TMB can be a relative value rather than an absolute value. In some embodiments, the subject's TMB status is compared to a control TMB value. In one embodiment, the subject's TMB status is within the highest quantile of the control TMB values. In another embodiment, the subject's TMB status is within the highest tertile of the control TMB values.

[0238] In some embodiments, TMB status is expressed as the number of mutations per sample, per cell, per exome, or per length of DNA (e.g., Mb). In some embodiments, a tumor has high TMB status if the tumor has at least about 50 mutations / tumor, at least about 55 mutations / tumor, at least about 60 mutations / tumor, at least about 65 mutations / tumor, at least about 70 mutations / tumor, at least about 75 mutations / tumor, at least about 80 mutations / tumor, at least about 85 mutations / tumor, at least about 90 mutations / tumor, at least about 95 mutations / tumor, at least about 100 mutations / tumor, at least about 105 mutations / tumor, at least about 110 mutations / tumor, at least about 115 mutations / tumor, or at least about 120 mutations / tumor. In certain embodiments, a tumor has high TMB status if the tumor has at least about 125 mutations / tumor, at least about 150 mutations / tumor, at least about 175 mutations / tumor, at least about 200 mutations / tumor, at least about 225 mutations / tumor, at least about 250 mutations / tumor, at least about 275 mutations / tumor, at least about 300 mutations / tumor, at least about 350 mutations / tumor, at least about 400 mutations / tumor, or at least about 500 mutations / tumor. In one particular embodiment, a tumor has high TMB status if the tumor has at least about 100 mutations / tumor.

[0239] In some embodiments, the tumor is characterized in that the tumor has been genetically, e.g., genomically sequenced by a TMB assay, e.g., FOUNDATIONONE® CDX (商標)A tumor has high TMB status if it has at least about 5 mutations (mutations / tumor), at least about 6 mutations / tumor, at least about 7 mutations / tumor, at least about 8 mutations / tumor, at least about 9 mutations / tumor, at least about 10 mutations / tumor, at least about 11 mutations / tumor, at least about 12 mutations / tumor, at least about 13 mutations / tumor, at least about 14 mutations / tumor, at least about 15 mutations / tumor, at least about 20 mutations / tumor, at least about 25 mutations / tumor, at least about 30 mutations / tumor, at least about 35 mutations / tumor, at least about 40 mutations / tumor, at least about 45 mutations / tumor, at least about 50 mutations / tumor, at least about 75 mutations / tumor, or at least about 100 mutations / tumor per megabase of the genome sequenced by the assay. In certain embodiments, a tumor has high TMB status if it has at least about 5 mutations / tumor. In certain embodiments, a tumor has a high TMB status when the tumor has at least about 10 mutations / tumor. In some embodiments, a tumor has a high TMB status when the tumor has at least about 11 mutations / tumor. In some embodiments, a tumor has a high TMB status when the tumor has at least about 12 mutations / tumor. In some embodiments, a tumor has a high TMB status when the tumor has at least about 13 mutations / tumor. In some embodiments, a tumor has a high TMB status when the tumor has at least about 14 mutations / tumor. In certain embodiments, a tumor has a high TMB status when the tumor has at least about 15 mutations / tumor.

[0240] Because mutation numbers vary by tumor type and other methods (see Q4 and Q5), the numbers associated with "TMB high" and "TMB low" may differ by tumor type.

[0241] PD-L1 status TMB status can be used, alone or in combination with other factors, as a means of predicting tumor response to a combination therapy comprising (a) an anti-PD-1 antibody or anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In some embodiments, the tumor's TMB status alone is used to identify patients with tumors that are likely to respond to a combination therapy comprising (a) an anti-PD-1 antibody or anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In other embodiments, PD-L1 status and TMB status are used to identify patients with tumors that are likely to respond to a combination therapy comprising (a) an anti-PD-1 antibody or anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In certain embodiments, the tumor has less than 1% PD-L1 expression, e.g., less than 1% of tumor cells express PD-L1. In certain embodiments, the subject has a high TMB status (≧10 mutations (mut) / Mb) and a tumor PD-L1 expression level of less than 1%.

[0242] The PD-L1 status of a tumor in a subject can be determined prior to administering a composition described herein or using a method described herein. PD-L1 expression can be determined by methods known in the art.

[0243] To assess PD-L1 expression, in one embodiment, a test tissue sample can be obtained from a patient in need of treatment. In another embodiment, assessing PD-L1 expression can be performed without obtaining a test tissue sample. In certain embodiments, selecting a suitable patient includes (i) providing a test tissue sample, optionally obtained from a patient having a tumor derived from NSCLC, where the test tissue sample contains tumor cells and / or tumor-infiltrating inflammatory cells; and (ii) assessing the proportion of cells in the test tissue sample that express PD-L1 on their cell surface based on assessing that the proportion of cells in the test tissue sample that express PD-L1 on their cell surface is higher than a predetermined threshold.

[0244] However, it should be understood that in methods involving measuring PD-L1 expression in a test tissue sample, the step involving providing a test tissue sample obtained from a patient is an optional step. It should also be understood that in certain embodiments, the "measuring" or "assessing" step to identify or determine the number or percentage of cells in a test tissue sample that express PD-L1 on their cell surface is performed by a modified method of measuring PD-L1 expression, for example, by performing a reverse transcriptase polymerase chain reaction (RT-PCR) assay or an IHC assay. In other embodiments, no modified step is included, and PD-L1 expression is assessed, for example, by reviewing test results reported from a laboratory. In certain embodiments, the method steps up to and including assessing PD-L1 expression provide an intermediate result that can be provided to a physician or other health care provider for use in selecting appropriate candidates for a combination therapy comprising (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In certain embodiments, the step of providing the intermediate result is performed by a physician or other person operating under the physician's guidance. In other embodiments, these steps are performed by an independent laboratory or by an independent person, such as a laboratory technician.

[0245] In certain embodiments of the methods of the invention, the proportion of cells expressing PD-L1 is assessed by performing an assay to determine the presence of PD-L1 RNA. In further embodiments, the presence of PD-L1 RNA is determined by RT-PCR, in situ hybridization, or RNase protection. In other embodiments, the proportion of cells expressing PD-L1 is assessed by performing an assay to determine the presence of PD-L1 polypeptide. In further embodiments, the presence of PD-L1 polypeptide is determined by immunohistochemistry (IHC), enzyme-linked immunosorbent assay (ELISA), in vivo imaging, or flow cytometry. In some embodiments, PD-L1 expression is assayed by IHC. In all other embodiments of these methods, cell surface expression of PD-L1 is assayed using, for example, IHC or in vivo imaging.

[0246] Imaging technologies provide important tools in cancer research and treatment. Recent developments in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflectance imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), laser scanning confocal microscopy (LSCM), and multiphoton microscopy (MPM), may foresee further applications of these technologies in cancer research. Some of these molecular imaging systems enable clinicians not only to see where tumors are located in the body, but also to visualize the expression and activity of specific molecules, cells, and biological processes that affect tumor behavior and / or response to therapeutic agents (Condeelis and Weissleder, "In vivo imaging in cancer," Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Combining the sensitivity and resolution of PET with antibody specificity, immunoPET imaging allows for the monitoring and assaying of antigen expression, particularly in tissue samples (McCabe and Wu, "Positive progress in immunoPET—not just a coincidence," Cancer Biother. Radiopharm. 25(3):253-61 (2010); Olafsen et al., "ImmunoPET imaging of B-cell lymphoma using 124I-anti-CD20 scFv dimers (diabodies)," Protein Eng. Des. Sel. 23(4):243-9 (2010)). In certain embodiments of the methods of the invention, PD-L1 expression is assayed by immunoPET imaging. In certain embodiments of the methods of the invention, the proportion of cells in a test tissue sample that express PD-L1 is assessed by performing an assay to determine the presence of PD-L1 polypeptide on the surface of cells in the test tissue sample. In certain embodiments, the test tissue sample is an FFPE tissue sample.In other embodiments, the presence of PD-L1 polypeptide is determined by an IHC assay. In further embodiments, the IHC assay is performed using automated methods. In some embodiments, the IHC assay is performed using an anti-PD-L1 monoclonal antibody that binds to the PD-L1 polypeptide. In particular embodiments, the anti-PD-L1 monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and combinations thereof. See WO / 2013 / 173223, which is incorporated herein by reference in its entirety.

[0247] In one embodiment of the method of the present invention, an automated IHC method is used to assay the expression of PD-L1 on the cell surface in FFPE tissue samples, such as tissue samples taken from tumors derived from NSCLC. The presence of human PD-L1 antigen can be measured in a test tissue sample and a negative control sample (e.g., normal tissue) by contacting the test tissue sample and a negative control sample with a monoclonal antibody that specifically binds to human PD-L1 under conditions that allow the formation of a complex between the monoclonal antibody or a portion thereof and human PD-L1. In a specific embodiment, the test and control tissue samples are FFPE samples. The formation of a complex is then detected, where a difference in complex formation between the test sample and the negative control sample indicates the presence of human PD-L1 antigen in the sample. Various methods can be used to quantify PD-L1 expression.

[0248] In certain embodiments, the automated IHC method includes (a) deparaffinizing and rehydrating embedded tissue sections in an automated stainer; (b) retrieving antigens by heating to 110°C for 10 minutes using an antigen decloaking chamber and pH 6 buffer; (c) loading the reagents into the automated stainer; and then (d) neutralizing endogenous peroxidase in the tissue sample; blocking nonspecific protein binding sites on the slide; incubating the slide with a primary antibody; incubating with a post-primary blocking agent; incubating with NovoLink polymer; adding a chromogenic substrate to develop color; and then counterstaining with hematoxylin, all of which are included in the automated stainer.

[0249] To assess PD-L1 expression in tumor tissue samples, pathologists performed microscopic analysis of membrane PD-L1 expression in each field. +The number of tumor cells is counted, and the percentage of positive cells is estimated mentally, then averaged to obtain a final percentage value. Different staining intensities are defined as 0 / negative, 1+ / weak, 2+ / intermediate, and 3+ / strong. Generally, percentage values ​​are first assigned to 0 and 3+ buckets, and then the intermediate 1+ and 2+ intensities are considered. For highly heterogeneous tissues, the sample is divided into zones, each zone is scored separately, and then combined into a single set of percentage values. The percentages of negative and positive cells for different staining intensities are determined from each region, and the median value is assigned to each zone. A final percentage value is assigned to the tissue for each staining intensity category: negative, 1+, 2+, and 3+. The sum of all staining intensities is required to equal 100%. In one embodiment, the threshold number of cells required for PD-L1 positivity is at least about 100, at least about 125, at least about 150, at least about 175, or at least about 200 cells. In certain embodiments, the threshold number of cells required as PD-L1 positive is at least about 100 cells. In certain embodiments, a tumor sample must have at least 100 total tumor cells to be considered an evaluable sample for PD-L1 expression.

[0250] Staining is also assessed in tumor-infiltrating inflammatory cells, such as macrophages and lymphocytes. Often, macrophages serve as an internal positive control, as staining is seen in the majority of macrophages. While staining at 3+ intensity is not required, the absence of macrophage staining should be considered to rule out technical failure. Macrophages and lymphocytes are assessed for cell membrane staining, and all samples are scored as positive or negative for each cell category. Staining is also characterized according to the designation of immune cells inside / outside the tumor. "Inside" means that immune cells are present within the tumor tissue and / or on the border of the tumor area without being physically intercalated between tumor cells. "Outside" means that there is no physical association with the tumor, and immune cells are found in the periphery or adjacent tissues associated with connective tissue.

[0251] In certain embodiments of these scoring methods, samples are scored by two independent pathologists, and the scores are then combined. In certain other embodiments, the identification of positive and negative cells is scored using appropriate software.

[0252] The histoscore is used as a more quantitative measure of the IHC data. The histoscore is calculated as follows: Histology score = [(%tumor x1 (low intensity)) + (%tumor x2 (intermediate intensity)) + (%tumor x3 (high intensity)].

[0253] To determine the tissue score, the pathologist estimates the percentage of cells staining in each intensity category within the sample. Because expression of many biomarkers is heterogeneous, the tissue score is a more accurate representation of overall expression. Final tissue scores range from 0 (no expression) to 300 (maximal expression).

[0254] Another method to quantify PD-L1 expression in test tissue sample IHC is to determine the adjusted inflammation score (AIS), which is defined as the intensity of inflammation multiplied by the percentage of PD-L1 expression by tumor-infiltrating inflammatory cells (Taube et al., "Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape," Sci. Transl. Med. 4(127):127ra37 (2012)).

[0255] In one embodiment, the PD-L1 expression level of the tumor is at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100%. In another embodiment, the PD-L1 expression status of the tumor is at least about 1%. In other embodiments, the PD-L1 expression status of the tumor is at least about 5%. In some embodiments, the PD-L1 expression status of the tumor is at least about 10%. In one embodiment, the PD-L1 expression status of the tumor is at least about 25%. In a particular embodiment, the PD-L1 expression status of the tumor is at least about 50%.

[0256] As used herein, "PD-L1 positive" can be used interchangeably with "at least about 1% PD-L1 expression." Thus, in one embodiment, a PD-L1-positive tumor can exhibit at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of tumor cells expressing PD-L1 as measured by automated IHC. In certain embodiments, "PD-L1 positive" refers to the presence of at least 100 cells that express PD-L1 on their cell surface. In other embodiments, "PD-L1 positive" refers to at least one tumor cell expressing PD-L1 on its surface in a tumor sample comprising at least 100 total tumor cells.

[0257] In one embodiment, tumors derived from NSCLC that are PD-L1 positive and have high TMB may have a greater response than tumors with only high TMB, only positive PD-L1 expression, or neither to a combination therapy comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) a post-induction phase comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject after (1). In one embodiment, the tumor derived from NSCLC has PD-L1 expression of at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50%. In certain embodiments, tumors derived from NSCLC with PD-L1 expression ≥ 50% and high TMB status are more likely to respond to combination therapy of (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CLTA-4 antibody than tumors with only high TMB, only ≥ 50% PD-L1 expression, or neither.

[0258] In certain embodiments, tumors in subjects suitable for the combination therapies described herein do not express PD-L1 (less than 1%, less than 2%, less than 3%, less than 4%, or less than 5% membranous PD-L1). In certain embodiments, the methods of the invention are independent of PD-L1 expression.

[0259] MSI Status TMB status can be used alone or in combination with other factors, such as MSI status, as a means for predicting the response of tumors derived from NSCLC to the combination therapy described herein.In one embodiment, MSI status is part of TMB status.In other embodiments, MSI status is measured separately from TMB status.

[0260] Microsatellite instability (MSI) is a state of genetic hypermutability resulting from impaired DNA mismatch repair (MMR). The presence of MSI is phenotypic evidence of impaired MMR function. In most cases, the genetic basis of MSI tumor instability is inherited germline alterations in any of five human MMR genes: MSH2, MLH1, MSH6, PMS2, and PMS1. In certain embodiments, tumors derived from NSCLC (e.g., colon tumors) have high microsatellite instability (MSI-H) and at least one mutation in the MSH2, MLH1, MSH6, PMS2, or PMS1 genes. In other embodiments, subjects undergoing tumor treatment in the control group do not have microsatellite instability (MSS or MSI stability) and do not have mutations in the MSH2, MLH1, MSH6, PMS2, and PMS1 genes.

[0261] In one embodiment, a subject suitable for the combination therapy described herein has a high TMB status and an MSI-H tumor derived from NSCLC. As used herein, an MSI-H tumor refers to a tumor with at least about 30% or more unstable MSI biomarkers. In some embodiments, a tumor derived from NSCLC is MSI-H when germline alterations are detected in at least two, at least three, at least four, or at least five MMR genes. In other embodiments, a tumor derived from NSCLC is MSI-H when germline alterations are detected in at least 30% of five or more MMR genes. In some embodiments, germline alterations in MMR genes are measured by polymerase chain reaction. In other embodiments, a tumor derived from NSCLC is MSI-H when at least one protein encoded by a DNA MMR gene is not detected in the tumor. In some embodiments, at least one protein encoded by a DNA MMR gene is detected by immunohistochemistry.

[0262] The tumor of the present invention The present invention relates to a method for treating a subject suffering from a tumor. The tumor may be of any type or derived from any tumor type. In some embodiments, the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer, and melanoma. In some embodiments, the tumor is derived from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer, lung cancer, or melanoma. In some embodiments, the tumor is derived from small cell lung cancer (SCLC). In some embodiments, the tumor is derived from NSCLC. In certain embodiments, the NSCLC is squamous NSCLC. In other embodiments, the NSCLC is non-squamous NSCLC. In certain embodiments, the tumor is stage IV NSCLC.

[0263] In some embodiments, the tumor is progressive. In certain embodiments, the tumor is locally advanced. In certain embodiments, the tumor is metastatic. In certain embodiments, the tumor is refractory. In certain embodiments, the tumor is refractory to one or more previous therapies for treating the tumor. In certain embodiments, the tumor is refractory to one or more standard therapies for treating the tumor. In certain embodiments, at least one previous treatment comprises chemotherapy. In certain embodiments, at least one previous treatment comprises platinum-based chemotherapy. In other embodiments, the patient has not received a previous therapy for treating the tumor, e.g., the patient is naive. In some embodiments, the tumor is not refractory. In some embodiments, the tumor is recurrent.

[0264] NSCLC NSCLC is the leading cause of cancer deaths in the United States and worldwide, surpassing breast, colon, and prostate cancers combined. In the United States, 228,190 new cases of lung and bronchial cancer are diagnosed, and approximately 159,480 deaths are attributed to the disease (Siegel et al. (2014) CA Cancer J Clin 64(1):9-29). The majority of patients (approximately 78%) are diagnosed with progressive / recurrent or metastatic disease. Metastasis from lung cancer to the adrenal gland is common, with approximately 33% of metastatic patients having such metastasis. NSCLC therapy has shown gradually improved overall survival (OS), but benefit has plateaued (the average OS for advanced-stage patients is just 1 year). Nearly all of these patients progress after first-line therapy, and the 5-year survival rate is only 3.6% in the refractory group. Between 2005 and 2009, the relative 5-year survival rate for lung cancer in the United States was 15.9% (NCCN Guidelines®, Version 3.2014 - Non-Small Cell Lung Cancer, available at www.nccn.org / professionals / physician_gls / pdf / nscl.pdf, last accessed May 14, 2014).

[0265] The method of the present invention can treat NSCLC tumors at all stages. In certain embodiments, the tumor is derived from any stage of NSCLC. There are at least seven stages of NSCLC: latent (hidden), stage 0 (carcinoma remains intraepithelial), stage I, stage II, stage IIIA, stage IIIB, and stage IV. In the latent stage, cancer cannot be detected by imaging or bronchoscopy. In stage 0, cancer cells are found in the lining of the airways.

[0266] In one embodiment, the method of the present invention treats stage I non-squamous NSCLC. Stage I NSCLC is divided into stages IA and IB. In stage IA, the tumor is in the lungs only and is 3 centimeters or less. In stage IB, the cancer has not spread to the lymph nodes and one or more of the following is true: 1) the tumor is larger than 3 centimeters but 5 centimeters or less; 2) the cancer has spread to the main bronchi and is at least 2 centimeters below where the trachea joins the bronchi; 3) the cancer has spread to the innermost layer of the membrane covering the lung; or 4) part of the lung has collapsed or developed non-infectious pneumonia (lung inflammation) in the area where the trachea joins the bronchi.

[0267] In another embodiment, the method of the present invention treats stage II non-squamous NSCLC. Stage II NSCLC is divided into stage IIA and IIB. In stage IIA, cancer may or may not have spread to lymph nodes. If cancer has spread to lymph nodes, it must only have spread to lymph nodes on the same side of the chest as the tumor, and the lymph nodes with cancer are located within the lung or near the bronchi, and one or more of the following conditions apply: 1) the tumor is 5 centimeters or less; 2) the cancer has spread to the main bronchi and is at least 2 centimeters below where the trachea joins the bronchi; 3) the cancer has spread to the innermost layer of the membrane that covers the lung, or 4) a portion of the lung has collapsed or developed non-infectious pneumonia (lung inflammation) in the area where the trachea joins the bronchi. Also, if the cancer has not spread to the lymph nodes, the tumor is considered stage IIA and one or more of the following is true: 1) the tumor is larger than 5 centimeters but not larger than 7 centimeters; 2) the cancer has spread to the main bronchi and is at least 2 centimeters below where the trachea joins the bronchi; 3) the cancer has spread to the innermost layer of the membrane that covers the lung; or 4) part of the lung has collapsed or developed noninfectious pneumonia (inflammation of the lung) in the area where the trachea joins the bronchi. In stage IIB, the cancer may or may not have spread to the lymph nodes. If cancer has spread to lymph nodes, it has spread only to lymph nodes on the same side of the chest as the tumor, and the lymph nodes with cancer are within the lung or near the bronchi, and one or more of the following is true: 1) the tumor is larger than 5 centimeters but not larger than 7 centimeters; 2) cancer has spread to the main bronchus and is at least 2 centimeters below where the trachea joins the bronchus; 3) cancer has spread to the innermost layer of the membrane that covers the lung, or 4) part of the lung has collapsed or noninfectious pneumonia (inflammation of the lung) has developed in the area where the trachea joins the bronchi.A tumor is considered stage IIB if the cancer has not spread to the lymph nodes and one or more of the following is true: 1) the tumor is larger than 7 centimeters; 2) the cancer has spread to the main bronchus (at least 2 centimeters below where the trachea joins the bronchi), chest wall, diaphragm, or nerves that control the diaphragm; 3) the cancer has spread to the membrane around the heart or inside the chest wall; 4) the entire lung has collapsed or the patient has developed non-infectious pneumonia (inflammation of the lungs); or 5) there are one or more separate tumors in the same lobe of the lung.

[0268] In another embodiment, the method of the present invention treats stage III non-squamous NSCLC. Stage IIIA is divided into three sections. These three sections are based on 1) tumor size; 2) where the tumor is found; and 3) if there is cancer in the lymph nodes. In the first type of stage IIIA NSCLC, cancer has spread to lymph nodes on the same side of the chest as the tumor, and the cancerous lymph nodes are located near the breastbone or where the bronchi enter the lungs. Additionally, 1) the tumor can be of any size; 2) part of the lung (where the trachea joins the bronchi) or the entire lung has collapsed or has developed non-infectious pneumonia (inflammation of the lungs); 3) there are one or more separate tumors in the same lobe of the lung; and 4) the cancer has spread to any of the following: a) the main bronchi (excluding where the bronchi join), b) the chest well, c) the diaphragm and the nerves that control it, d) the membranes around the lungs or the membrane that lines the chest wall, or e) the membrane around the heart.In the second type of Stage IIIA NSCLC, cancer has spread to lymph nodes on the same side of the chest as the tumor, and the cancerous lymph nodes are within the lung or near the bronchi. Additionally, 1) the tumor can be of any size; 2) the entire lung has collapsed or has developed non-infectious pneumonia (inflammation of the lungs); 3) there are one or more separate tumors in any of the lobules of the lung that have cancer; and 4) the cancer may have spread to any of the following: a) the main bronchi (excluding where the bronchi join), b) the chest wall, c) the diaphragm and the nerves that control it, d) the membranes around the lungs or that line the chest wall, e) the heart or membranes that lead to the heart, f) the great blood vessels that lead to or from the heart, g) the trachea, h) the esophagus, i) the nerves that control the larynx (voice box), j) the sternum / chest bone or spine, or k) the carina (where the trachea joins the bronchi). In the third type of NSCLC, stage IIIA, the cancer has not spread to the lymph nodes, the tumor can be any size, and the cancer has spread to one of the following: a) the heart, b) the large blood vessels leading to or from the heart, c) the trachea, d) the esophagus, e) the nerves that control the larynx (voice box), f) the sternum or spine, g) the carina (where the trachea joins the bronchi).Stage IIIB is divided into two categories based on 1) the size of the tumor, 2) where the tumor is found, and 3) which lymph nodes contain cancer. In the first type of Stage IIIB NSCLC, cancer has spread to lymph nodes on the opposite side of the chest from the tumor. Additionally, 1) the tumor can be of any size; 2) part of the lung (where the trachea joins the bronchi) or the entire lung has collapsed or developed noninfectious pneumonia (lung inflammation); 3) there are one or more separate tumors in any of the lung lobules containing cancer; and 4) the cancer has spread to any of the following: a) the main bronchi; b) the chest wall; c) the diaphragm and the nerves that control it; d) the membranes around the lungs or the membranes that line the chest wall; e) the heart or the membranes surrounding it; f) the large blood vessels leading to or from the heart; g) the trachea; h) the esophagus; i) the nerves that control the larynx (voice box); j) the sternum or spine; or k) the carina (where the trachea joins the bronchi). In the second type of stage IIIB NSCLC, cancer has spread to lymph nodes on the same side of the chest as the tumor. The cancer-containing lymph nodes are located near the sternum or where the bronchi enter the lungs. Additionally, 1) the tumors can be of any size, 2) there are separate tumors in different lobes of the same lung, 3) the cancer has spread to any of the following: a) the heart, b) the large blood vessels leading to or from the heart, c) the trachea, d) the esophagus, e) the nerves that control the larynx (voice box), f) the sternum or spine, or g) the carina (where the trachea joins the bronchi).

[0269] In some embodiments, the method of the present invention treats stage IV non-squamous NSCLC. In stage IV NSCLC, tumors can be of any size, and cancer has spread to lymph nodes. In stage IV NSCLC, one or more of the following is true: 1) there is one or more tumors in both lungs; 2) cancer is present in the fluid surrounding the lungs or heart; and 3) cancer has spread to other parts of the body, such as the brain, liver, adrenal glands, kidneys, or bones.

[0270] In some embodiments, the subject is a never smoker. In certain embodiments, the subject is a former smoker. In one embodiment, the subject is a current smoker. In certain embodiments, the subject has squamous cell carcinoma cells. In certain embodiments, the subject has non-squamous cell carcinoma cells.

[0271] Standard cancer treatment In some embodiments, the methods of the present invention are used instead of standard therapy. In certain embodiments, standard therapy is used in combination with the methods described herein. Standard therapy for various cancers is well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), a coalition of 21 major cancer centers in the United States, has published the NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®), which provides detailed and up-to-date information on standard therapy for various cancers (see NCCN Guidelines®, 2014).

[0272] colorectal cancer In some embodiments, the combination therapy treats a cancer that is colorectal cancer. In some embodiments, the colorectal cancer is colon cancer. In other embodiments, the colorectal cancer is rectal cancer. In certain embodiments, the colorectal cancer has microsatellite instability (MSI) (see Pawlik et al., Dis. Markers 20(4-5): 199-206 (2004)). In other embodiments, the colorectal cancer has low microsatellite instability (MSI-L).

[0273] Colorectal cancer is the third most common type of cancer in both men and women in the United States (see http: / / www.cancer.gov / types / colorectal, last accessed December 9, 2015). Most colorectal cancers are adenocarcinomas. Colon cancer is described in five stages: stage 0 (carcinoma confined to the epithelium), stage I, stage II, stage III, and stage IV. Six standard therapies are used for colon cancer: 1) surgery, including local resection, resection of the colon with anastomosis, or resection of the colon with colostomy; 2) radiofrequency ablation; 3) cryosurgery; 4) chemotherapy; 5) radiation therapy; and 6) targeted therapy, including monoclonal antibodies and angiogenesis inhibitors. In some embodiments, the combination therapy of the present invention treats colon cancer in conjunction with standard therapy.

[0274] Rectal cancer is classified into five stages: stage 0 (carcinoma confined to the epithelium), stage I, stage II, stage III, and stage IV. Six standard therapies are used for rectal cancer: 1) surgery, including polypectomy, local excision, resection, radiofrequency ablation, cryosurgery, and pelvic exenteration; 2) radiation therapy; 3) chemotherapy; and 4) targeted therapy, including monoclonal antibody therapy. In some embodiments, the methods of the present invention treat rectal cancer in conjunction with standard therapy.

[0275] lung cancer In some embodiments, the combination therapy of the present invention treats tumors derived from lung cancer. In certain embodiments, the cancer is NSCLC. In some embodiments, the NSCLC has squamous histology. In other embodiments, the NSCLC has non-squamous histology.

[0276] NSCLC is the leading cause of cancer deaths in the United States and worldwide, surpassing breast, colon, and prostate cancers combined. In the United States, 228,190 new cases of lung and bronchial cancer are diagnosed, and approximately 159,480 deaths are attributed to the disease (Siegel et al. (2014) CA Cancer J Clin 64(1):9-29). The majority of patients (approximately 78%) are diagnosed with progressive / recurrent or metastatic disease. Metastasis from lung cancer to the adrenal gland is common, with approximately 33% of metastatic patients having such metastasis. NSCLC therapy has shown gradually improved overall survival (OS), but benefit has plateaued (the average OS for advanced-stage patients is just 1 year). Nearly all of these patients progress after first-line therapy, and the 5-year survival rate is only 3.6% in the refractory group. Between 2005 and 2009, the relative 5-year survival rate for lung cancer in the United States was 15.9% (NCCN Guidelines®, Version 3.2014 - Non-Small Cell Lung Cancer, available at www.nccn.org / professionals / physician_gls / pdf / nscl.pdf, last accessed May 14, 2014).

[0277] There are seven stages of NSCLC: occult non-small cell lung cancer, stage 0 (the carcinoma is confined to the epithelium), stage I, stage II, stage IIIA, stage IIIB, and stage IV. In some embodiments, the combination therapy of the present invention treats NSCLC in conjunction with standard therapy.

[0278] In addition, the method of the present invention can also be combined with surgery, radiation therapy (RT) and chemotherapy, which are three commonly used therapies for treating NSCLC patients.As a class, NSCLC is relatively insensitive to chemotherapy and RT compared to small cell carcinoma.Generally, for patients with stage I or II disease, surgical resection offers the best chance of cure, and the use of chemotherapy agents both preoperatively and postoperatively is increasing.RT can also be used as adjuvant therapy for patients with resectable NSCLC, as primary local therapy, or as palliative therapy for patients with refractory NSCLC.

[0279] In one embodiment, the subject is a patient with stage IV disease. Patients with stage IV disease who have a good performance status (PS) benefit from chemotherapy. Many drugs, including platinum-based drugs (e.g., cisplatin, carboplatin), taxanes (e.g., paclitaxel, albumin-bound paclitaxel, docetaxel), vinorelbine, vinblastine, etoposide, pemetrexed, and gemcitabine, are useful for stage IV NSCLC. Combinations using many of these drugs result in 1-year survival rates of 30% to 40%, which are superior to single agents. Specific targeted therapies for the treatment of advanced lung cancer have also been developed. For example, bevacizumab (AVASTIN®) is a mAb that inhibits vascular endothelial growth factor A (VEGF-A). Erlotinib (TARCEVA®) is a small molecule TKI of the epidermal growth factor receptor (EGFR). Crizotinib (XALKORI®) is a small molecule TKI that targets ALK and MET and is used to treat NSCLC in patients carrying a mutated ALK fusion gene. Cetuximab (ERBITUX®) is a mAb that targets EGFR.

[0280] In some embodiments, the methods of the present invention are used to treat subjects with squamous NSCLC. In certain embodiments, the methods of the present invention are used in combination with standard therapy. Patients with squamous NSCLC (accounting for up to 25% of all NSCLC) have particularly unmet needs due to the limited treatment options available after first-line (1L) therapy. Following progression on platinum-based doublet chemotherapy, single-agent chemotherapy is the standard of care, resulting in a median OS of approximately 7 months. Docetaxel remains the benchmark treatment for this therapy, although erlotinib is also available, albeit less frequently. Pemetrexed has also been shown to produce clinically comparable efficacy results and to have significantly fewer side effects than docetaxel in the second-line (2L) therapy of patients with advanced NSCLC (Hanna et al., J Clin Oncol 22:1589-97). Currently, no therapeutic agents are approved for use in the third-line (3L) or later stages of lung cancer. Pemetrexed and bevacizumab are not approved for squamous NSCLC, and targeted therapies have limited application. Recent development failures, such as the failure of Oncothyreon and Merck KgaA's STIMUVAX® to improve OS in a phase 3 trial, the failure of ArQule and Daiichi Sankyo Co., Ltd.'s c-Met kinase inhibitor tivantinib to meet its survival endpoint, the failure of Eli Lilly's ALIMTA® and Roche's AVASTIN® combination to improve OS in a late-stage trial, and the failure of Amgen and Takeda Pharmaceuticals' small molecule VEGF-R antagonist motesanib to meet its clinical endpoint in a late-stage trial, create unmet need in advanced lung cancer.

[0281] Certain aspects of the present invention relate to methods of administering a chemotherapeutic agent for a period shorter than the standard period of chemotherapy. In some embodiments, the predetermined standard period of chemotherapy is based on the standard treatment for a predetermined cancer type. In certain embodiments, the tumor is derived from NSCLC, and the chemotherapeutic agent is administered for a period shorter than the standard period for administering standard chemotherapy for the treatment of NSCLC, for example, stage IV NSCLC.

[0282] Standard therapies for various cancers are well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), an alliance of 21 major cancer centers in the United States, publishes the NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®), which provide detailed, up-to-date information on standard therapies for a wide variety of cancers (see NCCN Guidelines®, 2018, available at www.nccn.org / professionals / physician_gls / default.aspx, last accessed October 22, 2018, the entire contents of which are incorporated herein by reference).

[0283] Patients with squamous cell NSCLC (which accounts for up to 25% of all NSCLC cases) have particularly unmet medical needs due to the paucity of treatment options after first-line (1L) therapy. Following progression on platinum-based doublet chemotherapy, single-agent chemotherapy is the standard of care, resulting in a median OS of approximately 7 months. Docetaxel remains the benchmark treatment for this regimen, although erlotinib is also available, albeit less frequently. Pemetrexed has also been shown to produce clinically comparable efficacy results and to have significantly fewer side effects compared with docetaxel in second-line (2L) therapy for patients with advanced NSCLC (Hanna et al., J Clin Oncol 22:1589-97).

[0284] NCCN guidelines for the treatment of NSCLC with chemotherapy include, but are not limited to, treatments selected from the following: (i) cisplatin 75 mg / m on day 1 for non-squamous cell carcinoma every 21 days for 4 cycles; 2 + pemetrexed 500mg / m on day 1 2 (ii) carboplatin AUC 6 on day 1 and paclitaxel 200 mg / m on day 1 every 21 days for 4 cycles 2 and (iii) carboplatin AUC 5 and pemetrexed 500 mg / m on day 1 for non-squamous epithelia, every 21 days for 4 cycles. 2 See NCCN Guidelines Version 6.2018 Non-Small Cell Lung Cancer. Other standard chemotherapy regimens include: (iv) cisplatin 50 mg / m on days 1 and 8 every 28 days for 4 cycles. 2 , and vinorelbine 25 mg / m on days 1, 8, 15, and 22 2 (v) Cisplatin 100 mg / m on day 1 every 28 days for 4 cycles 2 and vinorelbine 30 mg / m on days 1, 8, 15, and 22. 2 (vi) Cisplatin 75-80 mg / m on day 1 every 21 days for 4 cycles 2 and vinorelbine 25–30 mg / m on days 1 and 8 2 (vii) cisplatin 100 mg / m on day 1 every 28 days for 4 cycles 2 and etoposide 100 mg / m on days 1–3. 2 (viii) cisplatin 75 mg / m on day 1 every 21 days for 4 cycles 2 and gemcitabine 1250 mg / m on days 1 and 8 2 (ix) Cisplatin 75 mg / m on day 1 every 21 days for 4 cycles 2 and docetaxel 75 mg / m on day 1. 2and (x) carboplatin AUC 5 on day 1 and gemcitabine 1000 mg / m on days 1 and 8 every 21 days for 4 cycles. 2 Thus, in some embodiments, the methods of the present invention include administering an induction phase comprising administering chemotherapy, for example, standard chemotherapy, for a period shorter than the standard period of the chemotherapy. In certain embodiments, this period is less than 4 cycles. In certain embodiments, this period is less than 3 cycles. In certain embodiments, this period is less than 2 cycles. In certain embodiments, this period is 2 cycles.

[0285] In some embodiments, the subject has undergone one or more prior therapies for the tumor. In certain embodiments, at least one prior therapy comprises a standard therapy for treating stage IV NSCLC or a tumor derived therefrom. In some embodiments, at least one prior therapy comprises surgery, radiation therapy, chemotherapy, immunotherapy, or any combination thereof. In some embodiments, at least one prior therapy comprises chemotherapy. In certain embodiments, the at least one prior therapy is selected from treatments comprising administration of an anticancer agent selected from the group consisting of a platinum agent (e.g., cisplatin, carboplatin), a taxane (e.g., paclitaxel, albumin-bound paclitaxel, docetaxel), vinorelbine, vinblastine, etoposide, pemetrexed, gemcitabine, bevacizumab (AVASTIN®), erlotinib (TARCEVA®), crizotinib (XALKORI®), cetuximab (ERBITUX®), and any combination thereof. In certain embodiments, the at least one prior therapy comprises a platinum-based doublet chemotherapy.

[0286] In some embodiments, the subject has experienced disease progression after at least one prior therapy. In certain embodiments, the subject has received at least two prior therapies, at least three prior therapies, at least four prior therapies, or at least five prior therapies. In certain embodiments, the subject has received at least two prior therapies. In one embodiment, the subject has experienced disease progression after receiving at least two prior therapies. In certain embodiments, the at least two prior therapies include a first prior therapy and a second prior therapy, wherein the subject has experienced disease progression after the first prior therapy and / or the second prior therapy, wherein the first prior therapy includes surgery, radiation therapy, chemotherapy, immunotherapy, or any combination thereof; and the second prior therapy includes surgery, radiation therapy, chemotherapy, immunotherapy, or any combination thereof. In some embodiments, the first prior therapy includes platinum-based doublet chemotherapy, and the second prior therapy includes single-agent chemotherapy. In certain embodiments, the single-agent chemotherapy includes docetaxel.

[0287] In some aspects of the present invention, the methods of the present invention further comprise administering an additional anti-cancer therapy. The additional anti-cancer therapy may include any therapy known in the art and / or any standard therapy for treating NSCLC or tumors derived therefrom, as described herein. In some embodiments, the additional anti-cancer therapy comprises surgery, radiation therapy, chemotherapy, additional immunotherapy, or any combination thereof. In some embodiments, the additional anti-cancer therapy comprises additional chemotherapy, including any chemotherapy described herein. In some embodiments, the additional anti-cancer therapy comprises additional immunotherapy. In some embodiments, the additional anti-cancer therapy comprises administration of an antibody or antigen-binding portion thereof that specifically binds to LAG3, TIGIT, TIM3, NKG2a, OX40, ICOS, MICA, CD137, KIR, TGFβ, IL-10, IL-8, B7-H4, Fas ligand, CXCR4, mesothelin, CD27, GITR, or any combination thereof.

[0288] In certain embodiments, the additional anti-cancer therapy is administered simultaneously with or after the administration of the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In some embodiments, the additional anti-cancer therapy is administered simultaneously with the administration of the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In some embodiments, the additional anti-cancer therapy is administered after the administration of the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In some embodiments, the additional anti-cancer therapy is administered simultaneously with and after the administration of the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In other embodiments, the additional anti-cancer therapy is administered between the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In certain embodiments, the additional anti-cancer therapy, the anti-PD-1 antibody (or anti-PD-L1 antibody), and / or the anti-CTLA-4 antibody, are combined into a single formulation. In other embodiments, the additional anti-cancer therapeutic agent, the anti-PD-1 antibody (or anti-PD-L1 antibody), and / or the anti-CTLA-4 antibody, are in separate formulations.

[0289] melanoma In some embodiments, the combination therapy treats tumors derived from melanoma. Melanoma is the most lethal form of skin cancer and is the fifth most common cancer diagnosis in men and the seventh most common cancer diagnosis in women. (See http: / / www.cancer.gov / types / skin, last checked on December 9, 2015). There are seven stages of melanoma: Stage 0 (intraepidermal melanoma), Stage I, Stage II, Stage III that can be removed by surgery, Stage III that cannot be removed by surgery, Stage IV, and recurrent melanoma. Five types of standard therapies are used: 1) surgery; 2) chemotherapy; 3) radiation therapy; and 4) biologic therapy (including interferon, interleukin 2 (IL-2), tumor necrosis factor (TNF) therapy and ipilimumab), and 5) targeted therapy (including signal transduction inhibitor therapy (e.g., vemurafenib, dabrafenib, and trametinib), oncolytic virus therapy, monoclonal antibody therapy (including pembrolizumab and nivolumab), and angiogenesis inhibitors). In some embodiments, the combination therapy described herein treats melanoma in conjunction with standard therapy.

[0290] ovarian cancer In certain embodiments, the combination therapy treats a tumor, wherein the tumor originates from ovarian cancer, fallopian tube cancer, and / or primary peritoneal cancer ("ovarian cancer"). In certain embodiments, the cancer is ovarian epithelial cancer. In other embodiments, the cancer is ovarian germ cell tumor. In yet other embodiments, the cancer is ovarian low malignant potential tumor. In some embodiments, ovarian cancer arises in tissues encompassing the ovaries, peritoneum, or fallopian tubes (see http: / / www.cancer.gov / types / ovarian / patient / ovarian-epithelial-treatment-pdq, last accessed December 9, 2015).

[0291] There are four stages of ovarian cancer: stage I, stage II, stage III and stage IV, which include early ovarian cancer, advanced ovarian cancer and recurrent or persistent ovarian cancer.There are four standard therapies used for patients with ovarian cancer, fallopian tube cancer and primary peritoneal cancer: 1) surgery (including hysterectomy, unilateral salpingo-oophorectomy, bilateral salpingo-oophorectomy, omentectomy and lymph node biopsy); 2) radiation therapy; 3) chemotherapy; and 4) targeted therapy (including monoclonal antibody therapy and poly(ADP-ribose) polymerase inhibitor).Biological therapy is also being tested for ovarian cancer.In some embodiments, the combination therapy of the present invention is used in conjunction with standard therapy to treat ovarian cancer.

[0292] There are four stages of ovarian germ cell tumors: stage I, stage II, stage III, and stage IV. Four types of standard therapies are used: 1) surgery (including unilateral salpingo-oophorectomy, total hysterectomy, bilateral salpingo-oophorectomy, and tumor debulking); 2) observation; 3) chemotherapy; and 4) radiation therapy. New treatment options under investigation include high-dose chemotherapy with bone marrow transplantation. In some embodiments, the combination therapy of the present invention treats ovarian germ cell tumors in conjunction with standard therapy.

[0293] There are three stages of ovarian low malignant potential tumor: 1) early stage (stage I and stage II), 2) late stage (stage III and stage IB), and 3) recurrence. Two types of standard therapies are used: 1) surgery (including unilateral salpingo-oophorectomy, bilateral salpingo-oophorectomy, total hysterectomy, partial oophorectomy, and omentectomy), and 2) chemotherapy. In some embodiments, the combination therapy of the present invention treats ovarian low malignant potential tumor in conjunction with standard therapy.

[0294] head and neck cancer In some embodiments, the combination therapy treats cancer, and the cancer is head and neck cancer. Head and neck cancer includes cancer of the oral cavity, pharynx, larynx, paranasal sinuses, nasal cavity, and salivary glands. Head and neck cancer usually develops in squamous cells lining the moist mucosal surface inside the head and neck (e.g., inside the mouth, nose, and throat). These squamous cell carcinomas are often referred to as head and neck squamous cell carcinoma. Head and neck cancer can also develop in the salivary glands, although salivary gland cancer is relatively rare. (See http: / / www.cancer.gov / types / head-and-neck / head-neck-fact-sheet, last accessed December 9, 2015). The treatment plan for an individual patient depends on many factors, including the exact location of the tumor, the stage of the cancer, and the patient's age and overall health. Treatment for head and neck cancer can include surgery, radiation therapy, chemotherapy, targeted therapy, or a combination of treatments. In some embodiments, the combination therapy of the present invention treats head and neck cancer in conjunction with targeted therapy.

[0295] Pharmaceutical Compositions and Dosages Therapeutic agents of the present invention can be formulated into compositions, such as pharmaceutical compositions comprising an antibody and / or cytokine and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. Preferably, carriers for antibody-containing compositions are suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), while carriers for antibody and / or cytokine-containing compositions are suitable for parenteral administration, such as oral administration. In some embodiments, subcutaneous injection is based on Halozyme Therapeutics' ENHANZE® drug delivery technology (see U.S. Patent No. 7,767,429, the entire contents of which are incorporated herein by reference). ENHANZE® uses a co-formulation of an antibody with a recombinant human hyaluronidase enzyme (rHuPH20), which removes traditional limitations on the amount of biologics and drugs that can be delivered subcutaneously via an extracellular matrix (see U.S. Patent No. 7,767,429). Pharmaceutical compositions of the invention may contain one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Thus, in some embodiments, pharmaceutical compositions of the invention may further comprise a recombinant human hyaluronidase enzyme, such as rHuPH20.

[0296] In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a fixed dose together with the anti-CTLA-4 antibody in a single composition. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose together with the anti-CTLA-4 antibody. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose together with the anti-CTLA-4 antibody in a single composition. In some embodiments, the ratio of the anti-PD-1 antibody or anti-PD-L1 antibody to the anti-CTLA-4 antibody is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140 , about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1 or about 2:1 (mg).

[0297] Although higher concentrations of nivolumab monotherapy, up to 10 mg / kg every 2 weeks, were achieved without reaching the maximum tolerated dose (MTD), significant toxicities reported in other trials of checkpoint inhibitors and antiangiogenic therapeutic agents (see, e.g., Johnson et al., 2013 ; Rini et al., 2011 ) support the selection of nivolumab doses below 10 mg / kg.

[0298] Treatment continues as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in certain embodiments, the administered dose of the anti-PD-1 antibody, anti-PD-L1 antibody, and / or anti-CTLA-4 antibody is significantly lower than the approved dose, i.e., subtherapeutic dose of the drug. The anti-PD-1 antibody, anti-PD-L1 antibody, and / or anti-CTLA-4 antibody may be administered at a dose that produces the highest efficacy as a monotherapy in clinical trials, for example, about 3 mg / kg nivolumab administered once every 3 weeks (Topalian et al., 2012a; Topalian et al., 2012), or at a significantly lower dose, i.e., subtherapeutic dose.

[0299] The dosage and frequency of administration vary depending on the half-life of the antibody in the subject. Generally, human antibodies have the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. The dosage and frequency of administration vary depending on whether the treatment is preventive or therapeutic. In preventive applications, relatively low dosages are generally administered over a long period of time at relatively long intervals. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, relatively high dosages are often required at relatively short intervals until the progression of the disease slows or stops, or preferably until the patient shows partial or complete improvement in disease symptoms. Thereafter, the patient can be administered a preventive regimen.

[0300] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to provide an amount of the active ingredient that is not excessively toxic to the patient and is effective to achieve the desired therapeutic response for a particular patient, composition, and administration method. The selected dosage level can vary depending on the activity of the particular composition of the present invention used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular composition used, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and factors well known in the medical field. The composition of the present invention can be administered by one or more routes of administration using one or more of a variety of methods well known in the art. As will be understood by those skilled in the art, the route and / or method of administration can vary depending on the desired results.

[0301] kit Also within the scope of the present invention are kits containing (a) a chemotherapeutic agent, (b) an anti-PD-1 antibody or anti-PD-L1 antibody, and (c) an anti-CLTA-4 antibody for therapeutic use. The kit typically includes a label and instructions indicating the intended use of the contents of the kit. The term label includes any written or recorded material that accompanies or is included with or otherwise contained in the kit. In one embodiment, the present invention provides a kit for treating a subject suffering from a tumor, e.g., a tumor resulting from NSCLC, the kit including: (a) a dose of carboplatin sufficient to administer a dose of AUC 6, and a 200 mg / m 2(b) paclitaxel at a dose of 200 mg / kg to 800 mg / kg; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody at a dose of 200 mg / kg to 1800 mg / kg; (c) an anti-CTLA-4 antibody at a dose of 0.1 to 10 mg / kg; (d) instructions for use of (a) carboplatin and paclitaxel; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody; and (c) an anti-CTLA-4 antibody in the methods described herein. In one embodiment, the present invention provides a kit for treating a subject suffering from a tumor, e.g., a tumor derived from NSCLC, comprising: (a) a dose of carboplatin sufficient to administer a dose of AUC 5, and 2 (b) pemetrexed at a dose of 200 mg / kg to 800 mg / kg; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody at a dose of 200 mg / kg to 1800 mg / kg; (c) an anti-CTLA-4 antibody at a dose of 0.1 to 10 mg / kg; (d) instructions for use of (a) carboplatin and pemetrexed; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody; and (c) an anti-CTLA-4 antibody in the methods described herein. In one embodiment, the present invention provides a kit for treating a subject suffering from a tumor, e.g., a tumor derived from NSCLC, comprising: (a) a dose of carboplatin sufficient to administer a dose of AUC 6, and 2 (b) pemetrexed at a dose of 200 mg / kg to 800 mg / kg; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody at a dose of 200 mg / kg to 1800 mg / kg; (c) an anti-CTLA-4 antibody at a dose of 0.1 to 10 mg / kg; (d) instructions for use of (a) carboplatin and pemetrexed; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody; and (c) an anti-CTLA-4 antibody in the methods described herein. In one embodiment, the present invention provides a kit for treating a subject suffering from a tumor, e.g., a tumor derived from NSCLC, comprising: (a) 75 mg / m 2 of cisplatin and 500 mg / m 2(b) pemetrexed at a dose of 200 mg / kg to 800 mg; (b) an anti-PD-1 antibody at a dose of 200 mg / kg to 1800 mg; (c) an anti-CTLA-4 antibody at a dose of 0.1 to 10 mg / kg; and (d) instructions for use of (a) cisplatin and pemetrexed; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody; and (c) an anti-CTLA-4 antibody in the methods described herein.

[0302] In certain preferred embodiments for treating human patients, the kit comprises an anti-human PD-1 antibody described herein, such as nivolumab or pembrolizumab. In certain preferred embodiments for treating human patients, the kit comprises an anti-human PD-L1 antibody described herein, such as atezolizumab, durvalumab, or avelumab. In certain preferred embodiments for treating human patients, the kit comprises an anti-human CTLA-4 antibody described herein, such as ipilimumab, tremelimumab, MK-1308, or AGEN-1884.

[0303] In some embodiments, the kit further comprises a cytokine or a variant thereof. In certain embodiments, the kit comprises (a) an anti-PD-1 antibody or an anti-PD-L1 antibody, (b) an anti-CTLA-4 antibody, and (c) a CD122 agonist.

[0304] In some embodiments, the kit further comprises a comprehensive genomic profiling assay as described herein. In some embodiments, the kit comprises FOUNDATIONONE® CDX (商標)In one embodiment, the kit further comprises instructions for administering (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody to a subject identified as having a high TMB status, e.g., a TMB status of at least about 10 mutations per tumor of the examined genome, according to the methods described herein. In another embodiment, the kit further comprises instructions for administering (a) an anti-PD-1 antibody or an anti-PD-L1 antibody, (b) an anti-CTLA-4 antibody, and (c) a cytokine, e.g., a CD122 agonist, to a subject identified as having a high TMB status, e.g., a TMB status of at least about 10 mutations per tumor of the examined genome, according to the methods described herein.

[0305] In certain embodiments, the kit further comprises an assay for detecting and / or measuring PD-L1 expression on the surface of tumor cells, hi certain embodiments, the kit further comprises the Dako PD-L1 IHC 28-8 pharmDx test for IHC staining of PD-L1 protein on the surface of tumor cells.

[0306] All documents cited above, as well as all documents cited herein, are hereby incorporated by reference in their entirety.

[0307] The following examples are provided to illustrate the invention and should not be construed as limiting. [Example]

[0308] Example Example 1: Study of the combination of nivolumab and ipilimumab as first-line therapy for the treatment of stage IV non-small cell lung cancer (NSCLC) A clinical trial is underway to test the safety and efficacy of the combination of nivolumab and ipilimumab as first-line therapy for stage IV NSCLC. Nivolumab is administered intravenously at 3 mg / kg over 30 minutes every 2 weeks, in conjunction with ipilimumab administered intravenously at 1 mg / kg over 30 minutes every 6 weeks, until progression, unacceptable toxicity, or other prespecified reasons. Treatment with nivolumab and ipilimumab may continue for up to 2 years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0309] the purpose The primary objectives of this trial are (i) to determine the objective response rate (ORR) in all treated patients with stage IV NSCLC who are PD-L1 positive (membrane staining in ≥1% of tumor cells) as determined by blinded independent central review per RECIST 1.1 and who receive the combination of nivolumab and ipilimumab as first-line therapy, and (ii) to determine the ORR in all treated patients with stage IV NSCLC who are PD-L1 negative (≤1%) as determined by blinded independent central review per RECIST 1.1 and who receive the combination of ipilimumab and nivolumab as first-line therapy.

[0310] Secondary objectives include: (i) assessment of ORR by blinded independent central review per RECIST 1.1 in all treated patients who received nivolumab plus ipilimumab combination therapy as first-line therapy; (ii) assessment of progression-free survival (PFS) by blinded independent central review per RECIST 1.1; (iii) assessment of overall survival; (iv) assessment of ORR, PFS, and OS by PD-L1 expression level; and (v) assessment of tumor mutation burden (TMB) using DNA from tumor biopsies as a potential biomarker for predicting the efficacy (e.g., ORR, PFS, and OS) of nivolumab plus ipilimumab combination therapy.

[0311] Exploratory objectives include: (i) to evaluate the safety, tolerability, pharmacokinetics, and immunogenicity of nivolumab plus ipilimumab combination therapy as first-line therapy, and (ii) to evaluate tumor inflammatory gene expression signatures using RNA from tumor biopsies as potential biomarkers predictive of efficacy (e.g., ORR, PFS, and OS) of nivolumab plus ipilimumab combination therapy.

[0312] Clinical trial design Approximately 300 subjects with stage IV NSCLC, including at least 120 PD-L1-positive subjects and at least 100 PD-L1-negative subjects, will be enrolled and treated with the combination therapy. Subjects may receive nivolumab 3 mg / kg every two weeks and ipilimumab 1 mg / kg every six weeks. On the day of injection, nivolumab will be administered first. The second injection, if scheduled, must be administered with ipilimumab and may begin at least 30 minutes after the completion of the nivolumab injection. Subjects may receive the combination therapy until disease progression or unacceptable toxicity occurs, or for up to two years.

[0313] Subjects are permitted to use topical, ophthalmic, intra-articular, intranasal, and inhaled corticosteroids (minimizing systemic absorption). Adrenal replacement steroid doses of prednisone greater than 10 mg per day are permitted. Short-term (less than 3 weeks) corticosteroid treatment is permitted for prophylaxis (e.g., contrast allergy) or treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions caused by contact allergens). Routine concomitant use of bisphosphonates and RANK-L inhibitors for the prevention or mitigation of skeletal-related events in patients with bone metastases is permitted if initiated before the first dose of investigational therapy. Prior palliative radiation therapy must be completed at least 2 weeks prior to treatment.

[0314] Inclusion / Exclusion Criteria Eligible subjects must have histologically confirmed stage IV NSCLC (according to the 7th International Conference on the Classification of Lung Cancer) with squamous or non-squamous cell carcinoma and measurable disease by CT or MRI per RECIST 1.1 criteria. Subjects must have no prior systemic therapy for stage IV disease. Prior chemoradiotherapy for locally advanced disease is permitted as long as the last dose of chemotherapy or radiotherapy (whichever was last administered) occurred at least 6 months prior to enrollment. Patients with locally advanced disease (stage IIIB, specifically those for whom no curative treatment is available) who have relapsed after concurrent chemoradiotherapy are eligible for enrollment. Prior adjuvant or neoadjuvant chemotherapy for early-stage lung cancer is permitted as long as it was completed at least 6 months prior to initiating study therapy. Subjects must have EGFR / ALK wild-type disease and an EXOG PS of 0 or 1. Subjects must have tumor tissue samples available for PD-L1 immunohistochemistry (IHC) testing.

[0315] Either formalin-fixed, paraffin-embedded (FFPE) tissue blocks or unstained tumor tissue sections, along with relevant pathology reports, must be submitted for biomarker evaluation before treatment. Tumor tissue samples may be fresh or archived, as long as they were collected within 6 months prior to enrollment and no systemic therapy (e.g., adjuvant or neoadjuvant chemotherapy) was administered after sample collection. Tissue samples must be obtained by core needle biopsy, excision biopsy, or resection biopsy. Fine needle biopsy or cytospin drainage of pleural fluids is not considered appropriate for biomarker review and randomization. Biopsies of bone lesions without a soft tissue component or decalcified bone tumor samples are also not acceptable.

[0316] Subjects with known EGFR mutations sensitive to available targeted inhibitor therapy (including, but not limited to, exon 19 deletion mutations and exon 21 [L858R] substitution mutations) will be excluded. All subjects with non-squamous cell carcinoma must be tested for EGFR mutation status. EGFR testing will be performed locally. Use of an FDA-approved or local health department-approved test is strongly recommended. Tests other than PCR or next-generation sequencing should be retested using PCR- or next-generation sequencing-based methods. Subjects with non-squamous histology with unknown or indeterminate EGFR status will be excluded.

[0317] Subjects with known ALK translocations that are sensitive to available targeted inhibitor therapy will be excluded. If ALK testing is performed at a local laboratory, use of an FDA-approved test is strongly recommended. Subjects with unknown or undetermined ALK status may be enrolled.

[0318] Subjects with untreated CNS metastases are excluded.Subjects are eligible if their CNS metastases have been adequately treated and the subject has returned to baseline neurological status (excluding residual signs or symptoms related to CNS treatment) for at least two weeks before the first dose.In addition, subjects must be off corticosteroids or be receiving a stable or tapered dose of prednisone at 10 mg or less per day (or equivalent) for at least two weeks before the first dose.

[0319] Subjects with carcinomatous meningitis or active known or suspected autoimmune disease will be excluded. Subjects with type 1 diabetes, hypothyroidism to a degree requiring hormone replacement, skin diseases not requiring systemic treatment (vitiligo, psoriasis, alopecia, etc.), or conditions not expected to recur in the absence of external triggers will be eligible for enrollment.

[0320] Subjects with conditions requiring systemic treatment with either corticosteroids (>10 mg prednisone equivalent per day) or other immunosuppressants within 14 days of initial treatment are excluded. Inhaled or topical steroids and adrenal replacement steroids >10 mg prednisone equivalent per day are permitted in the absence of active autoimmune disease.

[0321] Subjects with a history of screening failure for any anti-PD-L1 antibody or clinical trial of an anti-PD-L1 antibody due to PD-L1-negative status will also be excluded.

[0322] Clinical trial evaluation Subjects are assessed for PD-L1 expression on tumor cells and classified into four groups: PD-L1 positive, PD-L1 negative, PD-L1 ≥ 50%, and PD-L1 unquantifiable. PD-L1 status can be determined by IHC staining of PD-L1 protein in submitted tumor samples using the Dako PD-L1 IHC 28-8 pharmDx test. PD-L1 positivity is characterized by ≥ 1% tumor cell membrane staining in at least 100 evaluable tumor cells. PD-L1 negativity is characterized by < 1% tumor cell membrane staining in at least 100 evaluable tumor cells. PD-L1 ≥ 50% is characterized by ≥ 50% tumor cell membrane staining in at least 100 evaluable tumor cells and is a subset of all PD-L1 positive subjects. Subjects with unquantifiable PD-L1 were defined as having unquantifiable PD-L1 expression at baseline, likely due to insufficient tumor biopsies for IHC staining and analysis. Key efficacy and safety parameters will be summarized narratively for tumors in subjects with unquantifiable PD-L1 status.

[0323] Tumor assessments by CT or MRI during the trial may be performed starting 6 weeks (± 7 days) after the date of first dose and every 6 weeks (± 7 days) until week 48. After week 48, tumor assessments may be performed every 12 weeks (± 7 days) until evidence of disease progression or treatment is discontinued, whichever occurs later.

[0324] In subjects receiving treatment containing nivolumab and ipilimumab, treatment beyond initial RECIST 1.1-defined progression is permitted if there is investigator-assessed clinical benefit and the combination of nivolumab and ipilimumab is tolerated. Subjects receiving investigational therapy beyond investigator-assessed progression must also continue tumor assessments until such time as treatment is discontinued.

[0325] OS will be followed continuously while subjects are taking the study drug and will be contacted in person or by telephone every 3 months after subjects stop receiving the study drug.

[0326] endpoint The primary endpoints were (i) the objective response rate (ORR) in all treated patients with PD-L1-positive disease (≥1%) as determined by blinded independent central review per RECIST 1.1 in stage IV NSCLC receiving the combination of nivolumab and ipilimumab as first-line therapy; and (ii) the ORR in all treated patients with PD-L1-negative disease (≤1%) as determined by blinded independent central review per RECIST 1.1 in stage IV NSCLC receiving the combination of ipilimumab and nivolumab as first-line therapy.

[0327] ORR was based on blinded independent central review per RECIST 1.1 criteria. ORR is defined as the number of subjects with a confirmed best overall response (BOR) of CR or PR divided by the number of treated PD-L1-positive or -negative subjects or by the total number of treated subjects. BOR is defined as the best response designation recorded between baseline and either the date of objectively documented progression per RECIST 1.1 or the date of initiation of palliative local therapy or subsequent anticancer therapy (whichever occurs first).

[0328] PFS, as determined by blinded independent central review, is defined as the time from the date of first administration for a treated subject to the date of first documented tumor progression by blinded independent central review (based on RECIST 1.1) or death from any cause. Subjects who have not progressed or died may be censored on the date of their last evaluable tumor assessment. Subjects who have not had a tumor assessment during the study may be censored on the date of their first administration. Subjects who have started palliative local therapy or subsequent anticancer therapy without prior documented progression may be censored on the date of their last evaluable tumor assessment before the start of the palliative local therapy or subsequent anticancer therapy, whichever occurs first.

[0329] Secondary endpoints were (i) ORR per blinded independent central review per RECIST 1.1 in all treated patients receiving first-line nivolumab plus ipilimumab, (ii) PFS per blinded independent central review per RECIST 1.1, (iii) OS, (iv) ORR, PFS, and OS by PD-L1 expression level; and (v) overall tumor cell mutations and their association with ORR, PFS, and OS. OS was defined as the time between the date of first administration and the date of death from any cause.

[0330] Example 2: Clinical Trial of Nivolumab Plus Ipilimumab in Combination with Chemotherapeutic Agents as First-Line Therapy for the Treatment of Stage IV Non-Small Cell Lung Cancer (NSCLC) A clinical trial is underway to evaluate the safety and tolerability of a phased first-line combination of nivolumab, ipilimumab, and chemotherapy for stage IV NSCLC. Nivolumab is administered intravenously and ipilimumab is administered intravenously along with two cycles of histology-based platinum-doublet chemotherapy as induction therapy, followed by nivolumab and ipilimumab until disease progression or unacceptable toxicity. Treatment with nivolumab and ipilimumab may continue for up to two years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0331] Two cycles of histology-based platinum-doublet chemotherapy included carboplatin AUC6 + paclitaxel 200 mg / m for squamous histology 2 For non-squamous tissues, (i) carboplatin AUC5 or carboplatin AUC6 + pemetrexed 500 mg / m 2 or (ii) cisplatin 75 mg / m 2 + pemetrexed 500mg / m 2 Includes:

[0332] the purpose The primary objectives of this trial are (i) to determine the incidence of dose-limiting toxicities (DLTs) during the DLT evaluation period (within 9 weeks after first dose); and (ii) to determine the safety and tolerability of nivolumab and ipilimumab in combination with chemotherapy agents.

[0333] Secondary objectives were to assess ORR and PFS by investigator review using RECIST 1.1, and OS.

[0334] Exploratory objectives include: (i) to assess global health status using the EQ-5D descriptive system and visual analogue scale, and lung cancer symptoms, as measured by the Lung Cancer Symptom Score (LCSS) Average Symptom Burden Index (ASBI), in subjects receiving nivolumab in combination with ipilimumab plus chemotherapy; (ii) to evaluate tumor inflammatory gene expression signatures as potential predictive biomarkers of efficacy (such as ORR, PFS, and OS) of nivolumab plus ipilimumab in combination with chemotherapy using RNA from tumor biopsies; and (iii) to explore TMB as a potential predictive biomarker of efficacy (such as ORR, PFS, and OS) of nivolumab plus ipilimumab in combination with chemotherapy using DNA from tumor biopsies.

[0335] Clinical trial design A safety lead-in phase may be conducted to assess safe dose levels. Approximately 28 subjects (ensuring at least 22 DLT-evaluable subjects) will receive two cycles of induction chemotherapy and nivolumab plus ipilimumab (Figure 1). The starting dose of nivolumab will be 360 ​​mg every 3 weeks, and ipilimumab will be 1 mg / kg every 6 weeks. Nivolumab will be administered with ipilimumab, plus two cycles of histology-based platinum-doublet chemotherapy. For squamous histology, carboplatin AUC6 plus paclitaxel 200 mg / m 2 For non-squamous tissues, (i) carboplatin AUC5 or carboplatin AUC6 + pemetrexed 500 mg / m 2 , or (ii) cisplatin 75 mg / m 2 and pemetrexed 500 mg / m 2 is administered.

[0336] After two cycles of induction therapy, nivolumab will be administered intravenously over 30 minutes in combination with ipilimumab administered intravenously over 30 minutes until progression, unacceptable toxicity, or other protocol-specified reasons. Treatment with nivolumab and ipilimumab will continue for up to two years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0337] "Safety" is defined as 25% or less of the subjects experiencing DLT (i.e., 5 or fewer of the 22 subjects assessed for DLT experienced such an event).

[0338] A safety assessment will be conducted for the first 10 subjects after a minimum of 9 weeks of follow-up. If 20% or less of the first 10 subjects experience DLT (i.e., 2 or fewer subjects have such events), the regimen will be deemed safe, and enrollment in subsequent clinical trials using this combination may begin while the safety run-in phase is ongoing. If 20% or more of the first 10 subjects experience DLT, a full safety cohort will be evaluated before using this dose regimen in subsequent trials. If 25% or more of all DLT-evaluable subjects experience DLT, the protocol may be modified to evaluate different dose levels depending on the observed toxicities.

[0339] After the two cycles of induction therapy, nivolumab and ipilimumab will be continued until consent is withdrawn in the event of disease progression or unacceptable toxicity, or for up to two years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0340] During the safety run-in phase, subjects are permitted to use topical, ophthalmic, intra-articular, intranasal, and inhaled corticosteroids (while minimizing systemic absorption). Corticosteroid doses of 10 mg or more of prednisone per day are permitted. Short-term (less than 3 weeks) administration of corticosteroids for prophylaxis (e.g., contrast allergy) or treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions caused by contact allergens) is permitted. Routine concomitant use of bisphosphonates and RANK-L inhibitors for the prevention or mitigation of skeletal-related events in patients with bone metastases...

Claims

1. 1. A method for treating a tumor in a subject in need thereof, comprising: (1) an induction phase comprising administering a chemotherapeutic agent to the subject for a period shorter than the standard period of chemotherapy; and (2) (1) followed by an induction period comprising administering to the subject an antibody that specifically binds to PD-1 (an "anti-PD-1 antibody") or an antigen-binding portion thereof, or an antibody that specifically binds to PD-L1 (an "anti-PD-L1 antibody") or an antigen-binding portion thereof. A method comprising:

2. 1. A method of treating a tumor in a subject in need thereof, comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject, wherein prior to administering the anti-PD-1 antibody or anti-PD-L1 antibody, the subject undergoes an induction phase comprising chemotherapy for a period that is shorter than the standard period of the chemotherapeutic agent.

3. The method of claim 1 or 2, wherein the induction phase further comprises administering an anti-PD-1 antibody or an anti-PD-L1 antibody.

4. 4. The method of any one of claims 1 to 3, further comprising administering an antibody that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody") or an antigen-binding portion thereof.

5. The method of claim 4, wherein the anti-CTLA-4 antibody is administered during the induction phase.

6. The method of claim 4 or 5, wherein the anti-CTLA-4 antibody is administered after the induction period.

7. The method of any one of claims 4 to 6, wherein the anti-CTLA-4 antibody is administered before or after the anti-PD-1 antibody or anti-PD-L1 antibody.

8. The method of any one of claims 4 to 7, wherein the anti-CTLA-4 antibody is administered after chemotherapy.

9. The method of claim 3, wherein the anti-CTLA-4 antibody is administered simultaneously with the anti-PD-1 antibody or the anti-PD-L1 antibody.

10. 10. The method of any one of claims 1 to 9, wherein the chemotherapeutic agent is administered for fewer than 10 cycles, fewer than 9 cycles, fewer than 8 cycles, fewer than 7 cycles, fewer than 6 cycles, fewer than 5 cycles, fewer than 4 cycles, or fewer than 3 cycles.

11. 11. The method of any one of claims 1 to 10, wherein the chemotherapeutic agent is administered in 5 cycles, 4 cycles, 3 cycles, 2 cycles or 1 cycle.

12. 12. The method of any one of claims 1 to 11, wherein the chemotherapy agent is administered for less than 5 cycles.

13. 13. The method of claim 12, wherein the chemotherapy agent is administered in fewer than four cycles.

14. 14. The method of claim 13, wherein the chemotherapy agent is administered in less than three cycles.

15. 15. The method of claim 14, wherein the chemotherapy agent is administered in less than two cycles.

16. 16. The method of any one of claims 1 to 15, wherein the chemotherapy agent is administered in a maximum of two cycles.

17. 17. The method of any one of claims 1 to 16, wherein the chemotherapy comprises standard therapy.

18. 18. The method of any one of claims 1 to 17, wherein the chemotherapeutic agent comprises an alkylating agent, an antimetabolite, an anti-microtubule agent, a topoisomerase inhibitor, a cytotoxic antibiotic, or any combination thereof.

19. 19. The method of any one of claims 1 to 18, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent.

20. 20. The method of any one of claims 1 to 19, wherein the chemotherapeutic agent comprises cisplatin, oxaliplatin, carboplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, or any combination thereof.

21. 21. The method of any one of claims 1 to 20, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and a second agent.

22. 22. The method of any one of claims 1 to 21, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and paclitaxel.

23. 22. The method of any one of claims 1 to 21, wherein the chemotherapeutic agent comprises a platinum-based chemotherapeutic agent and pemetrexed.

24. 23. The method of any one of claims 1 to 22, wherein the chemotherapeutic agents comprise carboplatin and paclitaxel.

25. 24. The method of any one of claims 1 to 21 and 23, wherein the chemotherapeutic agents comprise carboplatin and pemetrexed.

26. 24. The method of any one of claims 1 to 21 and 23, wherein the chemotherapeutic agents comprise cisplatin and pemetrexed.

27. 27. The method of any one of claims 1 to 26, wherein the chemotherapeutic agent is a histology-based chemotherapeutic agent.

28. 28. The method of any one of claims 1 to 27, wherein the chemotherapeutic agent is administered about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, or about once every 6 weeks.

29. 29. The method of any one of claims 10 to 28, wherein each cycle is three weeks.

30. 30. The method of claim 29, wherein the chemotherapy agent is administered on day 1 of each three-week cycle.

31. Chemotherapy consisted of carboplatin AUC 6 and paclitaxel 200 mg / m on any one day of a 3-week cycle. ２ 31. The method of any one of claims 1 to 30, comprising administering

32. Chemotherapy consisted of carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m ２ 31. The method of any one of claims 1 to 30, comprising administering

33. Chemotherapy was cisplatin 75 mg / m ２ and pemetrexed 500 mg / m ２ 31. The method of any one of claims 1 to 30, comprising administering

34. 34. The method of any one of claims 1 to 33, wherein the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1.

35. 35. The method of any one of claims 1 to 34, wherein the anti-PD-1 antibody binds to the same epitope as nivolumab.

36. 36. The method of any one of claims 1 to 35, wherein the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof.

37. 37. The method of any one of claims 1 to 36, wherein the anti-PD-1 antibody comprises a heavy chain constant region of the human IgG1 or human IgG4 isotype.

38. 38. The method of any one of claims 1 to 37, wherein the anti-PD-1 antibody is nivolumab.

39. 38. The method of any one of claims 1 to 37, wherein the anti-PD-1 antibody is pembrolizumab.

40. 40. The method of any one of claims 1 to 39, wherein the anti-PD-1 antibody is administered at a dose range of 0.1 mg to 10.0 mg per kg of body weight once every 2 weeks, 3 weeks, or 4 weeks.

41. 41. The method of any one of claims 1 to 40, wherein the anti-PD-1 antibody is administered at a dose of 3 mg, 5 mg, or 10 mg per kg of body weight once every three weeks.

42. 40. The method of any one of claims 1 to 39, wherein the anti-PD-1 antibody is administered in a fixed dose.

43. 43. The method of claim 42, wherein the anti-PD-1 antibody is administered at a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg.

44. 44. The method of claim 42 or 43, wherein the anti-PD-1 antibody is administered in a fixed dose about once every week, about every two weeks, about every three weeks, or about every four weeks.

45. 45. The method of any one of claims 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks.

46. 45. The method of any one of claims 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 240 mg once about every two weeks.

47. 45. The method of any one of claims 1 to 39, 43 and 44, wherein the anti-PD-1 antibody is administered at a fixed dose of about 480 mg once about every four weeks.

48. 34. The method of any one of claims 1 to 33, wherein the anti-PD-L1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof.

49. 49. The method of any one of claims 1 to 33 and 48, wherein the anti-PD-L1 antibody comprises a heavy chain constant region of the human IgG1 isotype.

50. 50. The method of any one of claims 1 to 33, 48 and 49, wherein the anti-PD-L1 antibody cross-competes with an antibody selected from atezolizumab, durvalumab and avelumab for binding to human PD-L1.

51. The method of any one of claims 1 to 33 and 48 to 50, wherein the anti-PD-L1 antibody binds to the same epitope on human PD-L1 as an antibody selected from atezolizumab, durvalumab, and avelumab.

52. The method of any one of claims 33 and 48 to 51, wherein the anti-PD-L1 antibody is atezolizumab, durvalumab, or avelumab.

53. 53. The method of any one of claims 1 to 33 and 48 to 52, wherein the anti-PD-L1 antibody is administered at a dose ranging from 0.1 mg to 15.0 mg per kg of body weight once every two weeks, three weeks, or four weeks.

54. 54. The method of any one of claims 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered at a dose of 3 mg or 5 mg per kg of body weight once every two weeks.

55. 54. The method of any one of claims 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered at a dose of 10 mg / kg body weight once every three weeks.

56. 54. The method of any one of claims 1 to 33 and 48 to 53, wherein the anti-PD-L1 antibody is administered in a fixed dose.

57. 57. The method of claim 56, wherein the anti-PD-Ll antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, at least about 1400 mg, or at least about 1500 mg.

58. 58. The method of claim 56 or 57, wherein the anti-PD-L1 antibody is administered in a fixed dose once every week, every two weeks, every three weeks, or every four weeks.

59. 60. The method of any one of claims 1 to 33, 48 to 53, 57 and 58, wherein the anti-PD-L1 antibody is administered at a flat dose of about 1200 mg once about every three weeks.

60. 60. The method of any one of claims 1 to 59, wherein the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or portion thereof.

61. 61. The method of any one of claims 1 to 60, wherein the anti-CTLA-4 antibody comprises a heavy chain constant region of the human IgG1 isotype.

62. 62. The method of any one of claims 1 to 61, wherein the anti-CTLA-4 antibody is ipilimumab.

63. 63. The method of any one of claims 1 to 62, wherein the anti-CTLA-4 antibody is tremelimumab.

64. 64. The method of any one of claims 1 to 63, wherein the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

65. 65. The method of any one of claims 1 to 64, wherein the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg to at least about 10.0 mg per kg of body weight once about every week, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, or about every 12 weeks.

66. 66. The method of any one of claims 1 to 65, wherein the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg of body weight about once every six weeks.

67. 66. The method of any one of claims 1 to 65, wherein the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg of body weight once about every 12 weeks.

68. 68. The method of any one of claims 1 to 67, wherein the anti-CTLA-4 antibody is administered in a fixed dose.

69. (i) an anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; and (ii) the anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; 69. The method of any one of claims 1 to 68.

70. (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents are carboplatin AUC 6 and paclitaxel 200 mg / m ２ administered on day 1 of an approximately 3 week cycle; 69. The method of any one of claims 1 to 68.

71. (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents are carboplatin AUC 5 and pemetrexed 500 mg / m ２ administered on day 1 of an approximately 3 week cycle; 70. The method of any one of claims 1 to 69.

72. (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapy agents are carboplatin AUC 6 and pemetrexed 500 mg / m ２ administered on day 1 of an approximately 3 week cycle; 70. The method of any one of claims 1 to 69.

73. (i) the anti-PD-1 antibody is administered at a fixed dose of about 360 mg once about every three weeks; (ii) an anti-CTLA-4 antibody is administered at a dose of about 1 mg per kg of body weight once about every six weeks; and (iii) The chemotherapeutic agent is cisplatin 75 mg / m ２ and pemetrexed 500 mg / m ２ administered on day 1 of an approximately 3 week cycle; 70. The method of any one of claims 1 to 69.

74. 74. The method of any one of claims 70 to 73, wherein the chemotherapeutic agent is administered in less than three cycles.

75. 75. The method of any one of claims 70 to 74, wherein the chemotherapy agent is administered over two cycles.

76. 76. The method of any one of claims 1 to 75, wherein the subject exhibits a progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

77. 77. The method of any one of claims 1 to 76, wherein the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

78. 78. The method of any one of claims 1 to 77, wherein the subject exhibits a response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.

79. 79. The method of any one of claims 1 to 78, wherein the tumor has a high tumor mutational burden (TMB) status.

80. 80. The method of claim 79, wherein the TMB status is determined by sequencing nucleic acid in the tumor and identifying genomic alterations in the sequenced nucleic acid.

81. Genome modification, (i) one or more somatic mutations; (ii) one or more nonsynonymous mutations; (iii) one or more missense mutations; (iv) one or more alterations selected from the group consisting of base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and combinations thereof; or (v) any combination of (i) to (iv) 81. A composition for use in the method of claim 80 comprising:

82. High TMB is at least 210, at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 82. The composition for use in the method of any one of claims 79 to 81, having a score of at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495 or at least 500.

83. A composition for use in the method of any one of claims 79 to 82, wherein the subject's TMB status is compared to a control TMB value, and wherein the subject's TMB status is within the highest quantile of the control TMB values ​​or the subject's TMB status is within the highest tertile of the control TMB values.

84. 84. A composition for use in the method of any one of claims 79 to 83, wherein the biological sample comprises a tumor tissue biopsy, a liquid biopsy, blood, serum, plasma, exoRNA, circulating tumor cells, ctDNA, cfDNA, or any combination thereof.

85. TMB state, (i) genome sequencing, (ii) exome sequencing; (iii) genomic profiling, or (iv) Any combination of (i) to (iii).

85. A composition for use in the method of any one of claims 79 to 84, determined by

86. ゲノムプロファイルが FAKE1, FAKE 1, FAKE1 2、SYS3、SYS2、SH142、SYS1 0、SIC4、SIC2、SIC1、SIC2、DAM2 、KHY4、HYH3、KHYS1、SHYS、SYS1 1、SHA1K、SHASH2、SHASH、SHASH、SHASH、 WHY、DYS、DYSYS、SYSYS、SYS1 、DY4、DYSYS、DYSY、DY4(S173). 9)、SYSYS(SYS3)、SYSYSYS、SYS1、SYS3 3、SYS、SYS2、SYS1、SYSYS、SYS 1, HYS, SYS2, SYS, S, S, S, S, S, S, S, S, S CHR3、CH11、CHYCH2、CHY11、CHY 56、SIGN、DYSHY、SYSHY、SYS3、DYS TH、SYSY、SYSY、SYS13、SYSYS、SYSY、1 SHY、SYS1、SYS、SYS1(SYS123)、 11030000000000000000000000000000000000000000+ JY、BYSHY、SYSYS1、SYSYプロモーターのみ)、HARSH、HARSH11、SHARH11 DASH、SHA1、SH1H、SHASH1、SHASH2 、SHE、SIR、SHERS10、SEE10、SHEY124、SEE 4、DYS(HYS1)、SHICH2、SHY2 、DAY、DAY3、DAY14、DAY2Y 、KHYK、SHYS2、SHYSHY、SYSHYS3、SHYS1 、KHY1、CHHYH、CHH19、HHH3、HHHHH、N9 88、DYS2、SHOSE、SHOSES14、SHOSE1S 002、SHYS、CHY23、CHY3、CHY22(5) 、D1、DYS1、SYS2、SYS1、SYS10、S DA3、DY2、CHY3、CHY3、CHY20043) 、DY2、DYS、SYS1、SYS22、SYS2、SYS 1. CHRIC1、FEN4、FENSE、CHE200042)、9 CHEY22、SYSY12、SYS2、SYS53、SYSY1、 DA274、DYS3、CHS6、DN12、SHAS、9 CHEEK、SHEY1、SHEY2、SHEY1、SHEY、SHEYSYS、DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR, ATRX, CDC7 3, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AUR KA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARCB1, VEGFA, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WISP3, AXL, C DK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, E RBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BARD1, CDKN1 A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1 B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BCL2L1, CDKN 2A, ERG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CD KN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, CDKN2C, E 86. The composition of claim 85, comprising one or more genes selected from the group consisting of SR1, FUBP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RBI, STAT3, and combinations thereof.

87. (i) the tumor comprises non-small cell carcinoma; (ii) the tumor is relapsed or refractory after at least one prior choice of therapy for treating the tumor; or (iii) Both (i) and (ii); 87. A composition for use in the method of any one of claims 79 to 86.

88. Genomic profile is FOUNDATIONNONE® CDX （商標） 86. The composition of claim 85, comprising:

89. 89. A composition for use in the method of claim 79 or 80 or a composition described in any one of claims 81 to 88, wherein the tumor has a TMB of at least about 10 mutations per megabase of the sequenced genome.

90. 90. The method or composition of any one of claims 1 to 89, wherein the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer and melanoma.

91. 90. The method or composition of any one of claims 1 to 89, wherein the tumor is derived from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer or melanoma.

92. 92. The method or composition of any one of claims 1 to 91, wherein the tumor is derived from non-small cell lung cancer (NSCLC) or small cell lung cancer.

93. 93. The method or composition of any one of claims 1 to 92, wherein the tumor is derived from NSCLC.

94. 94. The method or composition of any one of claims 1 to 93, wherein the tumor is derived from stage IV NSCLC.

95. 95. The method or composition of claim 93 or 94, wherein the NSCLC is squamous NSCLC.

96. 95. The method or composition of claim 93 or 94, wherein the NSCLC is non-squamous NSCLC.

97. 97. The method or composition of any one of claims 1 to 96, wherein the tumor is locally advanced, progressive or metastatic.

98. 98. The method or composition of any one of claims 1 to 97, wherein the tumor is refractory or recurrent.

99. 99. The method or composition of any one of claims 1-98, wherein the tumor is refractory after at least one prior therapy for treating said tumor, wherein the at least one prior therapy comprises a standard therapy.

100. 100. The method or composition of claim 99, wherein the at least one prior therapy comprises prior chemotherapy.

101. 101. The method or composition of claim 100, wherein the prior chemotherapy is a platinum-based chemotherapy.

102. 102. The method or composition of any one of claims 1 to 101, wherein at least 1% of the tumor cells exhibit membrane PD-L1 expression.

103. 103. The method or composition of any one of claims 1 to 102, wherein at least 5% of the tumor cells exhibit membrane PD-L1 expression.

104. 104. The method or composition of any one of claims 1 to 103, wherein the anti-PD-1 antibody and the chemotherapeutic agent are administered on the same day.

105. 104. The method or composition of any one of claims 1 to 103, wherein the anti-PD-1 antibody and the chemotherapeutic agent are administered on different days.

106. 105. The method or composition of any one of claims 1 to 104, wherein the dose of anti-PD-1 antibody, the dose of anti-CTLA-4 antibody, and the dose of chemotherapeutic agent are all administered on the same day.

107. (i) the anti-PD-1 antibody is administered once every three weeks; (ii) an anti-CTLA-4 antibody is administered once every six weeks; and (iii) the chemotherapy agent is administered once every three weeks for two cycles; wherein the dose of anti-PD-1 antibody, the dose of anti-CTLA-4 antibody, and the dose of chemotherapeutic agent are all administered on day 1 of a first 3-week cycle.

105. The method or composition of any one of claims 1 to 104.

108. 108. The method or composition of claim 107, wherein the dose of anti-PD-1 antibody and the dose of chemotherapeutic agent are administered on day 1 of a second 3-week cycle.

109. 109. The method or composition of claim 107 or 108, wherein the dose of anti-PD-1 antibody and the dose of anti-CTLA-4 antibody are administered on day 1 of a third 3-week cycle.

110. 110. The method or composition of any one of claims 1 to 109, wherein the period between the first administration after the induction phase and the last administration of the induction phase is equal to or less than about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days (2 weeks), about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days (1 month), about 31 days (1 month), about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, or about 3 months.

111. 111. The method or composition of any one of claims 1-98 and 102-110, wherein the tumor is not refractory to chemotherapy during or after the induction period.

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