Methods to treat tumors

Abstract

The present disclosure provides a method for treating a subject suffering from a tumor, for example, derived from non-small cell lung cancer (NSCLC), comprising administering (a) chemotherapy, (b) a combination of an anti-PD-1 antibody or an anti-PD-L1 antibody, and (c) an anti-CTLA-4 antibody to the subject, wherein the chemotherapy is administered for a period shorter than the standard duration.

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] This PCT application claims the benefit of priority of U.S. provisional application No. 62 / 749,393 filed on October 23, 2018, the full text of which is incorporated herein by reference.

[0003] Field of the present disclosure

[0004] The present disclosure provides a method for treating a subject suffering from a tumor using immunotherapy combined with chemotherapy. Background Technology

[0005] Immunotherapy approaches have recently demonstrated clinical efficacy in several cancer types, including melanoma and hormone-refractory prostate cancer. Tumors can modulate and evade host immune responses through multiple mechanisms, including the downregulation of tumor-specific antigen expression and presentation, the secretion of anti-inflammatory cytokines, and the upregulation of inhibitory ligands. T cell checkpoint regulators, such as CTLA-4 and programmed death-1 (PD-1, CD279), are cell surface molecules that induce signaling cascades that downregulate T cell activation and proliferation when bound by their cognate ligands.

[0006] PD-1 is a major immune checkpoint receptor expressed by activated T and B cells and mediates immunosuppression. PD-1 is a member of the CD28 family of receptors, which includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1, programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2), have been identified; these are expressed in antigen-presenting cells as well as many human cancers and have been found to downregulate T cell activation and cytokine secretion upon binding to PD-1. 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).

[0007] Nivolumab (previously designated as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that blocks the down-regulation of anti-tumor T-cell function by selectively blocking interactions with PD-1 ligands (PD-L1 and PD-L2) (U.S. Patent No. 8,008,449; Wang et al., 2014). Nivolumab has demonstrated activity in various advanced solid tumors, including renal cell carcinoma (renal adenocarcinoma or adrenal tumor), melanoma, and non-small cell lung cancer (NSCLC) (Topalian et al., 2012a; Topalian et al., 2014; Drake et al., 2013; WO 2013 / 173223).

[0008] The combination of immunotherapies with different mechanisms of action offers the potential for synergistic responses. Although PD-1 and CTLA-4 are both co-inhibitor molecules, there is evidence suggesting that they utilize distinct mechanisms to limit T-cell activation. Ipilimumab and anti-CTLA-4 antibodies have been shown to enhance the anticancer activity of nivolumab.

[0009] However, there remains a need to further enhance the efficacy of anti-PD-1 or anti-PD-L1 therapy. The present disclosure describes a method of administering a combination of an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a platinum-based dual chemotherapy.

[0010] A specific aspect of the present disclosure relates to a method for treating a tumor in a subject requiring treatment of a tumor, comprising: (1) an induction step comprising administering chemotherapy to a subject requiring treatment of a tumor for a period shorter than the standard duration for said chemotherapy; and (2) a post-induction step comprising administering to the subject after (1) an antibody that specifically binds to PD-1 or its antigen-binding portion ("anti-PD-1 antibody") or an antibody that specifically binds to PD-L1 or its antigen-binding portion ("anti-PD-L1 antibody").

[0011] Some aspects of the present disclosure relate to a method for treating a tumor in a subject requiring treatment of a tumor, comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to a subject requiring treatment of a tumor, wherein, prior to administering the anti-PD-1 antibody or the anti-PD-L1 antibody, an induction step comprising administering chemotherapy to the subject for a period shorter than the standard period for said chemotherapy.

[0012] In some embodiments, the induction step 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 or its antigen-binding portion ("anti-CTLA-4 antibody"). In some embodiments, the anti-CTLA-4 antibody is administered during the induction step. In some embodiments, the anti-CTLA-4 antibody is administered during the post-induction step. In some embodiments, the anti-CTLA-4 antibody is administered before or after the anti-PD-1 antibody or the 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 co-administered with the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0013] In some embodiments, chemotherapy 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, chemotherapy is administered for 5 cycles, 4 cycles, 3 cycles, 2 cycles, or 1 cycle. In some embodiments, chemotherapy is administered for fewer than 5 cycles. In some embodiments, chemotherapy is administered for fewer than 4 cycles. In some embodiments, chemotherapy is administered for fewer than 3 cycles. In some embodiments, chemotherapy is administered for fewer than 2 cycles. In some embodiments, chemotherapy is administered for 2 cycles or less.

[0014] In some embodiments, chemotherapy includes standard management therapy. In some embodiments, chemotherapy includes alkylating agents, and an antimetabolites, antimicrotubules, topoisomerase inhibitors, cytotoxic antibiotics, or any combination thereof.

[0015] In some embodiments, the chemotherapy includes platinum-based chemotherapy. In some embodiments, the chemotherapy includes cisplatin, oxaliplatin, carboplatin, nedaplatin, triflatin tetranitrate, phenantriplatin, picoplatin, satraplatin, or any combination thereof. In some embodiments, the chemotherapy includes platinum-based chemotherapy and a second agent. In some embodiments, the chemotherapy includes platinum-based chemotherapy and paclitaxel. In some embodiments, the chemotherapy includes platinum-based chemotherapy and pemetrexed. In some embodiments, the chemotherapy includes carboplatin and paclitaxel. In some embodiments, the chemotherapy includes carboplatin and pemetrexed. In some embodiments, the chemotherapy includes cisplatin and pemetrexed. In some embodiments, the chemotherapy is histology-based.

[0016] In some embodiments, chemotherapy is administered once every approximately 2 weeks, once every approximately 3 weeks, once every approximately 4 weeks, once every approximately 5 weeks, or once every approximately 6 weeks. In some embodiments, each cycle is 3 weeks. In some embodiments, chemotherapy is administered on the 1st day of each 3-week cycle.

[0017] In some embodiments, chemotherapy consists of carboplatin AUC 6 and paclitaxel 200 mg / m² 2 It includes administering on the first day of each 3-week cycle. In some embodiments, the chemotherapy consists of carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m² 2 It includes administering. In some embodiments, the chemotherapy is cisplatin 75 mg / m² 2 and pemetrexed 500 mg / m² 2 It includes administering.

[0018] 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 its antigen-binding portion. In some embodiments, the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 isoform or a human IgG4 isoform. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the anti-PD-1 antibody is pembrolizumab.

[0019] In some embodiments, the anti-PD-1 antibody is administered once every 2, 3, or 4 weeks at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered once every 3 weeks at a dose of 3 mg / kg, 5 mg / kg, or 10 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered at a uniform dose. In some embodiments, the anti-PD-1 antibody is administered in a uniform 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 once every about 1, 2, 3, or 4 weeks in a uniform dose. In some embodiments, the anti-PD-1 antibody is administered once every about 3 weeks in a uniform dose of about 360 mg. In some embodiments, the anti-PD-1 antibody is administered once every two weeks at a uniform dose of about 240 mg. In some embodiments, the anti-PD-1 antibody is administered once every four weeks at a uniform dose of about 480 mg.

[0020] In some embodiments, the anti-PD-L1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or its antigen-binding portion. In some embodiments, the anti-PD-L1 antibody comprises a heavy chain constant region of a human IgG1 isoform. In some embodiments, the anti-PD-L1 antibody cross-competes for binding to human PD-L1 with an antibody selected from atezolizumab, durvalumab, and avelumab. 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.

[0021] In some embodiments, the anti-PD-L1 antibody is administered once every 2, 3, or 4 weeks at a dose ranging from 0.1 mg / kg to 15.0 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every 2 weeks at a dose of 3 mg / kg or 5 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every 3 weeks at a dose of 10 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose. In some embodiments, the anti-PD-L1 antibody is administered in a uniform 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 once every approximately 1, 2, 3, or 4 weeks at a uniform dose. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1200 mg.

[0022] In some embodiments, the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or a part thereof. In some embodiments, the anti-CTLA-4 antibody comprises a heavy chain constant region of a human IgG1 isoform. 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.

[0023] In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 12 weeks at a dose of about 3 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered at a uniform dose.

[0024] In some embodiments, (i) an anti-PD-1 antibody is administered once every 3 weeks at a uniform dose of about 360 mg, and (ii) an anti-CTLA-4 antibody is administered once every 6 weeks at a dose of about 1 mg / kg body weight.

[0025] In some embodiments, (i) an anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 360 mg; (ii) an anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight; and (iii) chemotherapy is carboplatin AUC 6 and paclitaxel 200 mg / m² 2 It includes administering on the first day of each 3-week cycle.

[0026] In some embodiments, (i) an anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 360 mg; (ii) an anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight; and (iii) chemotherapy is carboplatin AUC 5 and pemetrexed 500 mg / m² 2 It includes administering on the first day of each 3-week cycle.

[0027] In some embodiments, (i) an anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 360 mg; (ii) an anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight; and (iii) chemotherapy is carboplatin AUC 6 and pemetrexed 500 mg / m² 2 It includes administering on the first day of each 3-week cycle.

[0028] In some embodiments, (i) an anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 360 mg; (ii) an anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight; and (iii) chemotherapy is cisplatin 75 mg / m² 2 and pemetrexed 500 mg / m² 2 It includes administering on the first day of each 3-week cycle.

[0029] In some embodiments, chemotherapy is administered for less than 3 cycles. In some embodiments, chemotherapy is administered for 2 cycles.

[0030] In some embodiments, the subject exhibits progression-free survival for 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 some embodiments, the subject exhibits a total 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 some embodiments, the object exhibits an objective response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0031] In some embodiments, the tumor has a TMB state with a high tumor mutation burden (TMB). In some embodiments, the TMB state is determined by sequencing nucleic acids within the tumor and identifying genomic alterations within the sequenced nucleic acids. In some embodiments, 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 changes (CNAs), gene rearrangements, and any combination thereof; or (v) any combination of (i)-(iv).

[0032] In some embodiments, the 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 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 It has a score of 490, at least 495, or at least 500.

[0033] In some embodiments, the TMB state of the object is compared with a reference TMB value, wherein the TMB state of the object is within the highest quartile of the reference TMB value or wherein the TMB state of the object is within the upper quartile of the reference TMB value.

[0034] 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.

[0035] In some embodiments, the TMB status is determined by (i) genome sequencing, (ii) exome sequencing, (iii) genome profiling, or (iv) any combination of (i)-(iii). In some embodiments, the genome profile is 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 (C17orf39), 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), C11orf30 (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, PLCG2, 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, 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, CDC73, 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, 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, SPENIt comprises one or more genes selected from the group consisting of ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, 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 any combination thereof.

[0036] In some embodiments, (i) the tumor comprises a non-small cell carcinoma, (ii) the tumor is relapsed or refractory after at least one prior regimen to treat the tumor, or (iii) both (i) and (ii).

[0037] In some embodiments, the genome profile includes FOUNDATIONONE® CDX™. In some embodiments, the tumor has a TMB of at least about 10 mutations per megabase of the sequenced genome.

[0038] In some embodiments, the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastric cancer, esophageal cancer, bladder cancer, lung cancer, and melanoma. In some embodiments, the tumor originates from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastric cancer, esophageal cancer, bladder cancer, lung cancer, or melanoma. In some embodiments, the tumor originates from non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC). In some embodiments, the tumor originates from NSCLC. In some embodiments, the tumor originates from stage IV NSCLC. In some embodiments, the NSCLC is squamous NSCLC. In some embodiments, the NSCLC is non-squamous NSCLC.

[0039] In some embodiments, the tumor is locally advanced, progressive, or metastatic. In some embodiments, the tumor is refractory or relapsed. In some embodiments, the tumor is refractory after at least one prior therapy to treat the tumor, wherein at least one prior therapy includes standard management therapy. In some embodiments, at least one prior therapy includes prior chemotherapy. In some embodiments, the prior chemotherapy is platinum-based chemotherapy.

[0040] In some embodiments, at least 1% of the tumor cells express membrane PD-L1. In some embodiments, at least 5% of the tumor cells express membrane PD-L1.

[0041] In some embodiments, the anti-PD-1 antibody and chemotherapy are administered on the same day. In some embodiments, the anti-PD-1 antibody and chemotherapy 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 chemotherapy are all administered on the same day.

[0042] In some embodiments, (i) an anti-PD-1 antibody is administered once every 3 weeks, (ii) an anti-CTLA-4 antibody is administered once every 6 weeks, and (iii) chemotherapy is administered once every 3 weeks for 2 cycles; wherein the dose of the anti-PD-1 antibody, the dose of the anti-CTLA-4 antibody, and the dose of chemotherapy are all administered on the 1st day of the first 3-week cycle. In some embodiments, the dose of the anti-PD-1 antibody and the dose of chemotherapy are administered on the 1st day 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 the 1st day of the third 3-week cycle.

[0043] In some embodiments, the period between the first administration of the post-induction phase and the last administration of the induction phase is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days (2 weeks), 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days (1 month), 31 days (1 month), 5 weeks, 6 weeks, 7 weeks, 8 weeks, 2 months, or 3 months or less.

[0044] In some embodiments, the tumor is not refractory to chemotherapy during or after the induction phase.

[0045] Other features and advantages of the present disclosure will become apparent from the following detailed description and examples, and should not be construed as limiting. The contents of all cited references, including scientific papers, newspaper reports, GenBank registrations, patents, and patent applications cited throughout this application, are expressly incorporated herein by reference. Brief explanation of the drawing

[0046] Figure 1 shows the study design for a safety induction study to evaluate safe dose levels for nivolumab and ipilimumab administered with histology-based platinum dual chemotherapy lead-in treatment. NSCLC = Non-small cell lung cancer; SQ = Squamous; NSQ = Non-squamous; DLT = Dose-limiting toxicity. Figure 2 shows the study design for a clinical trial evaluating the safety and efficacy of treating stage IV NSCLC using introduction therapy including a combination of nivolumab and ipilimumab and histology-based platinum dual chemotherapy. SQ = flat; NSQ = non-flat. Specific details for implementing the invention

[0047] The present disclosure provides a method for treating a subject suffering from a tumor, comprising: (1) an induction step comprising administering chemotherapy to a subject suffering from a tumor for a period shorter than the standard duration for said chemotherapy; and (2) a post-induction step comprising administering to the subject after (1) an antibody that specifically binds to PD-1 or its antigen-binding portion (“anti-PD-1 antibody”) or an antibody that specifically binds to PD-L1 or its antigen-binding portion (“anti-PD-L1 antibody”). The present specification further provides a method for treating a tumor in a subject suffering from a tumor, comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to a subject suffering from a tumor, wherein, prior to administering the anti-PD-1 antibody or the anti-PD-L1 antibody, an induction step comprising administering chemotherapy to the subject for a period shorter than the standard duration for said chemotherapy is present. In some embodiments, the tumor originates from NSCLC.

[0048] terminology

[0049] To facilitate a better understanding of the present disclosure, specific terms are defined first. Unless otherwise explicitly provided herein, each of the following terms used in this application will have the meaning set forth below. Additional definitions are provided throughout the application.

[0050] "Administering" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of the various methods and delivery systems known to a person skilled in the art. Preferred routes of administration for immunotherapy, e.g., anti-PD-1 antibody or anti-PD-L1 antibody, include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral administration routes, e.g., by injection or infusion. As used herein, the term "parenteral administration" means a mode of administration, typically by injection, other than enteral and local administration, and includes, without limitation, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transarticular, subcutaneous, interarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. Other parenteral routes include oral, topical, epidermal, or mucosal administration routes, e.g., intranasal, vaginal, rectal, sublingual, or topical. Additionally, administration may be performed, e.g., once, multiple times, and / or over one or more extended periods.

[0051] As used herein, “Adverse Event” (AE) is any undesirable, generally unintended, or unwanted sign (including abnormal laboratory findings), symptom, or disease associated with the use of a medical treatment. For example, an adverse event may be associated with the activation of the immune system or the expansion of immune system cells (e.g., T cells) occurring in response to treatment. A medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity. Reference to a method that may “change an adverse event” means a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0052] The "antibody" (Ab) will, without limitation, comprise a glycoprotein immunoglobulin or its antigen-binding portion comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds, which specifically bind to an antigen. Each H chain is a heavy chain variable region (V in this application). H It includes the heavy chain invariant region (abbreviated as ). The heavy chain invariant region consists of three invariant domains, C H1 , C H2 and C H3 Each light chain includes a light chain variable region (V in this application). L It includes (abbreviated as ) and a light chain invariant region. The light chain invariant region consists of one invariant domain, C L Includes. V H and V L The region can be further subdivided into a hypermutability region referred to as the Complementarity Decision Region (CDR), which is interspersed with more conserved regions referred to as the Framework Region (FR). Each V H and V L It consists of three CDRs and four FRs arranged in the following order from amino-terminus to carboxy-terminus: 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 immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the typical complement system.

[0053] Immunoglobulins may be derived from any commonly known isoforms, including but not limited to IgA, secreted IgA, IgG, and IgM. IgG subclasses are also widely known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. “Isoform” refers to a class or subclass of antibodies (e.g., IgM or IgG1) encoded by a heavy chain constant region gene. The term “antibody” includes, for example, both naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; fully synthetic antibodies; and single-chain antibodies. Non-human antibodies may be humanized by recombinant methods so as to reduce their immunogenicity in humans. Unless explicitly stated otherwise, and unless otherwise indicated by the context, the term “antibody” also includes antigen-binding fragments or antigen-binding portions of any of the aforementioned immunoglobulins, monovalent and divalent fragments or portions, and single-chain antibodies.

[0054] "Isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities (for example, an isolated antibody that specifically binds to PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1). However, an isolated antibody that specifically binds to PD-1 may have cross-reactivity with other antigens, such as PD-1 molecules from different species. Furthermore, the isolated antibody may be substantially free of other cellular materials and / or chemicals.

[0055] The term "monoclonal antibody" (mAb) refers to a non-naturally occurring preparation of an antibody molecule of single molecular composition, that is, an antibody molecule in which the primary sequence is essentially identical and exhibits single-binding specificity and affinity for a specific epitope. Monoclonal antibodies are examples of isolated antibodies. Monoclonal antibodies may be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0056] "Human antibody" (HuMAb) refers to an antibody having a variable region in which both the framework and the CDR region are derived from a human wiring immunoglobulin sequence. Additionally, if the antibody contains a constant region, the constant region is also derived from a human wiring immunoglobulin sequence. The human antibodies of this disclosure may contain amino acid residues not encoded by the human wiring immunoglobulin sequence (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation). However, the term "human antibody" as used herein is not intended to include antibodies in which a CDR sequence derived from the wiring of another mammalian species, e.g., mouse, is grafted onto a human framework sequence. The terms "human antibody" and "fully human antibody" are used synonyms.

[0057] "Humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDR of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulin. In one embodiment of the humanized form of the antibody, some, most, or all of the amino acids outside the CDR are replaced with amino acids from human immunoglobulin, while some, most, or all of the amino acids within the CDR remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are acceptable, provided that they do not eliminate the antibody's ability to bind to a specific antigen. The "humanized antibody" retains antigen specificity similar to that of the original antibody.

[0058] "Chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, for example, an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0059] "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.

[0060] The "antigen-binding portion" (also called "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen bound by the whole antibody.

[0061] "Cancer" refers to a broad group of various diseases characterized by the uncontrolled growth of abnormal cells within the body. Uncontrolled cell division and growth lead to the formation of malignant tumors that divide and grow to invade neighboring tissues, and can also metastasize to distal parts of the body via the lymphatic system or bloodstream.

[0062] The term “immunotherapy” refers to treating a subject who has a disease, is at risk of developing the disease, or is experiencing a recurrence of the disease by means of a method including inducing, enhancing, suppressing, or otherwise modifying an immune response. “Treatment” or “therapy” of a subject refers to any type of intervention or course performed on a subject, or the administration of an activator therefor, for the purpose of reversing, alleviating, improving, suppressing, slowing, or preventing the onset, progression, development, severity, or recurrence of symptoms, complications, or conditions associated with the disease, or biochemical indications.

[0063] "Programmed Death-1" (PD-1) refers to an immunosuppressive receptor belonging to the CD28 family. PD-1 is predominantly expressed on previously 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, and analogs having at least one common epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank accession number U64863.

[0064] "Programmed death ligand-1" (PD-L1) is one of two cell surface glycoprotein ligands for PD-1 that downregulate T cell activation and cytokine secretion upon binding to PD-1 (the other is PD-L2). As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, and analogs having at least one common epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank accession number Q9NZQ7.

[0065] "Cytotoxic T-lymphocyte antigen-4" (CTLA-4) refers to an immunosuppressive 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 referred to as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), variants, isoforms, and species homologs of hCTLA-4, and analogs having at least one common epitope with hCTLA-4. The complete hCTLA-4 sequence can be found under GenBank accession number AAB59385.

[0066] "Subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates, e.g., non-human primates, sheep, dogs, and rodents, e.g., mice, rats, and guinea pigs. In a preferred embodiment, the subject is a human. The terms "subject," "patient," and "participant" are used interchangeably herein.

[0067] In relation to the methods and dosages of the present disclosure, the use of the term “uniform dose” means a dose administered to a patient regardless of the patient’s body weight or body surface area (BSA). Accordingly, the uniform dose is not provided as a mg / kg dose, but rather as an absolute amount of the agent (e.g., anti-PD-1 antibody). For example, a 60 kg person and a 100 kg person will receive the same dose of antibody (e.g., 360 mg of anti-PD-1 antibody).

[0068] In connection with the methods of the present disclosure, the use of the term “fixed dose” means that two or more different antibodies (e.g., anti-PD-1 antibody and anti-CTLA-4 antibody or anti-PD-L1 antibody and anti-CTLA-4 antibody) are present in the composition in a specific (fixed) ratio relative to each other. In some embodiments, the fixed dose is based on the weight of the antibody (e.g., mg). In certain embodiments, the fixed dose is based on the concentration of the antibody (e.g., mg / ml). In some embodiments, the ratio 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, approximately 60:1, approximately 50:1, approximately 40:1, approximately 30:1, approximately 20:1, approximately 15:1, approximately 10:1, approximately 9:1, approximately 8:1, approximately 7:1, approximately 6:1, approximately 5:1, approximately 4:1, approximately 3:1, or approximately 2:1 mg of first antibody (e.g., anti-PD-1 antibody or anti-PD-L1 antibody) to mg of second antibody (e.g., anti-CTLA-4 antibody). For example, a 3:1 ratio of anti-PD-1 antibody and anti-CTLA-4 antibody may mean that the vial may contain approximately 240 mg of anti-PD-1 antibody and 80 mg of anti-CTLA-4 antibody or approximately 3 mg / ml of anti-PD-1 antibody and 1 mg / ml of anti-CTLA-4 antibody.

[0069] The term "weight-based dose" as used herein means that the dose administered to a patient is calculated based on the patient's body 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 (i.e., 180 mg) can be calculated and used for administration.

[0070] The “therapeutic effective dose” or “therapeutic effective dosage” of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with another therapeutic agent, protects the subject from the onset of the disease or promotes disease regression, as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or the prevention of damage or disability caused by disease suffering. The ability of a therapeutic agent to promote disease regression may be evaluated using various methods known to a skilled physician, for example, by testing the activity of the agent in human subjects during clinical trials, in animal model systems predicting efficacy in humans, or in in vitro assays.

[0071] For example, an "anticancer agent" promotes cancer regression in a subject. In a preferred embodiment, a therapeutically effective dose of the drug promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administering an effective dose of the drug alone or in combination with an antineoplastic agent causes a reduction in tumor growth or size, necrosis of the tumor, a reduction in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free periods, or the prevention of damage or disability caused by disease suffering. Additionally, the terms "effective" and "effectiveness" in relation to treatment include both pharmacological efficacy and physiological safety. Pharmacological efficacy refers to the ability of the drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (adverse effects) at the cellular, organ, and / or organism level resulting from the administration of the drug.

[0072] As an example of the treatment of a tumor, e.g., a tumor derived from NSCLC, a therapeutically effective dose of anticancer agent inhibits cell growth or tumor growth by preferably at least about 20%, more preferably at least about 40%, more preferably at least about 60%, and more preferably at least about 80% compared to an untreated subject. In another preferred embodiment of the present disclosure, tumor regression may be observed and continued for a period of at least about 20 days, more preferably at least about 40 days, or more preferably at least about 60 days. Despite these ultimate measures of therapeutic efficacy, the evaluation of an immunotherapy drug must also take into account immune-related response patterns.

[0073] "Immune response" is understood in the relevant technical field and generally refers to a biological response within a vertebrate against foreign agents or abnormal cells, e.g., cancerous cells, which protects the organism against these agents or the disease induced by them. The immune response is mediated by the action of one or more types of 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, which cause the selective targeting, binding to, damage to, and / or removal from the vertebrate body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in the case of autoimmune or pathological inflammation, normal human cells or tissues. The immune response, e.g., T cells, e.g., effector T cells, Th cells, CD4 + Cell, CD8 + It includes the activation or inhibition of T cells or Treg cells, or the activation or inhibition of any other cells of the immune system, such as NK cells.

[0074] "Immune-related response patterns" refer to clinical response patterns often observed in cancer patients treated with immunotherapies that generate antitumor effects by inducing cancer-specific immune responses or by modifying natural immune processes. These response patterns are characterized by beneficial therapeutic effects followed by an early increase in tumor burden or the appearance of new lesions, which are classified as disease progression and synonymous with drug failure in the evaluation of traditional chemotherapy. Therefore, the proper evaluation of immunotherapies may require long-term monitoring of the effects of these agents on the target disease.

[0075] "Immunomodulator" or "immunomodulator" refers to an agent that targets, for example, a component of a signaling pathway that may be involved in the coordination, regulation, or modification of an immune response. "Coordination," "regulation," or "modification" of an immune response refers to any change in the activity of immune system cells or of these cells (e.g., effector T cells, e.g., Th1 cells). Such coordination includes stimulation or inhibition 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 changes that may occur within the immune system. Both inhibitory and stimulating immunomodulators have been identified, some of which may have enhanced functions in the tumor microenvironment. In some embodiments, immunomodulators target molecules on the surface of T cells. "Immunomodulatory target" or "immunomodulatory target" is a molecule, for example, a cell surface molecule, that is targeted for binding by a substance, agent, moiety, compound, or molecule, and whose activity is altered by such binding. Immunomodulatory targets include, for example, receptors ("immunomodulatory receptors") and receptor ligands ("immunomodulatory ligands") on the cell surface.

[0076] “Immunotherapy” refers to treating a subject who has a disease, is at risk of developing a disease, or is experiencing a recurrence of a disease by means of a method comprising inducing, enhancing, suppressing, or otherwise modifying the immune system or immune response. In certain embodiments, immunotherapy involves administering antibodies to the subject. In other embodiments, immunotherapy involves administering small molecules to the subject. In other embodiments, immunotherapy involves administering cytokines or their analogs, variants, or fragments.

[0077] "Immunostimulatory therapy" or "immunostimulant therapy" refers to a therapy that induces (induces or enhances) an immune response in a subject, for example, to treat cancer.

[0078] "Enhancing the endogenous immune response" means increasing the efficacy or potency of the existing immune response in the subject. Such an 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.

[0079] The therapeutic effective dose of the drug includes a “preventive effective dose,” which is any amount of the drug that inhibits the occurrence or recurrence of cancer when administered alone or in combination with an antineoplastic agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant state) or a subject at risk of experiencing cancer recurrence. In a preferred embodiment, the preventive effective dose completely prevents the occurrence or recurrence of cancer. “Inhibiting” the occurrence or recurrence of cancer means reducing the likelihood of cancer occurrence or recurrence, or completely preventing cancer occurrence or recurrence.

[0080] As used herein, the term “induction phase” refers to a part 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 includes administering chemotherapy to the subject. In some embodiments, the induction phase includes administering chemotherapy and immunotherapy to the subject, for example, a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody or an anti-PD-L1 antibody and an anti-CTLA-4 antibody. In some embodiments, the induction phase includes administering chemotherapy for a period shorter than the standard duration for that chemotherapy. In some embodiments, chemotherapy is administered for less than 4 cycles. In some embodiments, chemotherapy is administered for less than 3 cycles. In some embodiments, chemotherapy is administered for less than 2 cycles. In some embodiments, chemotherapy is administered for 2 cycles. In some embodiments, the induction phase is (i) standard management chemotherapy modified to reduce the duration of chemotherapy administration; and (ii) administering 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 for about 6 weeks. In certain embodiments, the induction phase lasts for 2 cycles, each cycle being 21 days.

[0081] As used herein, the term “post-induction phase” refers to any period during therapy or treatment that occurs after the induction phase. The post-induction phase may continue for any amount of time. In some embodiments, the post-induction phase continues until disease progression, an unacceptable adverse event, a complete response, or up to two years. In some embodiments, the post-induction phase includes immunotherapy comprising the administration of 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 the administration of chemotherapy. In certain embodiments, the post-induction phase begins on the first day in which immunotherapy is administered in the absence of chemotherapy.

[0082] As used herein, the term "tumor mutation burden" (TMB) refers to the number of somatic mutations within the tumor genome and / or the number of somatic mutations per genomic region of the tumor. Gland (hereditary) variants are excluded when determining the TMB because the immune system is more likely to recognize them as self. Tumor mutation burden (TMB) may also be used interchangeably with "tumor mutation load," "burden of tumor mutations," or "burden of tumor mutations."

[0083] TMB is a genetic analysis of the tumor genome and can therefore be measured by applying sequencing methods widely known to those skilled in the art. By comparing tumor DNA with DNA from patient-matched normal tissue, wiring mutations or polymorphisms can be eliminated.

[0084] In some embodiments, TMB is determined by sequencing tumor DNA using high-throughput sequencing technology, e.g., next-generation sequencing (NGS) or NGS-based methods. In some embodiments, NGS-based methods are selected from whole-genome sequencing (WGS), whole-exome sequencing (WES), or comprehensive genome profiling (CGP) of cancer gene panels, e.g., FoundationOne CDX™ and MSK-IMPACT clinical trials. In some embodiments, as used herein, TMB 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, e.g., missense mutations (i.e., altering a specific amino acid within a protein) and / or nonsense mutations (causing premature termination and subsequent truncation of the protein sequence), identified by normalizing the matched tumor to a wiring sample and excluding any hereditary wiring gene alterations. In another embodiment, TMB is measured using the total number of missense mutations in the tumor. 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 NsM per megabase (NsM / Mb). 1 megabase represents 1 million bases.

[0085] The TMB status can be a numerical or relative value, e.g., high, medium, or low; it can be within the highest quantile of the reference set or within the upper tranche.

[0086] As used herein, the term "high TMB" refers to the number of somatic mutations within the genome of a tumor that exceeds the number of normal or average somatic mutations. In some 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 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, Having a score of 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, the high TMB has a score of 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 a specific embodiment, the high TMB has a score of at least 243.

[0087] In another embodiment, “high TMB” refers to a TMB within the highest quantile of the reference TMB value. For example, all subjects with evaluable TMB data are grouped according to the quantile distribution of TMB, that is, subjects are ranked from the highest number of genetic alterations to the lowest number and divided into a defined number of groups. In one embodiment, all subjects with evaluable TMB data are ranked and divided into thirds, and “high TMB” is within the upper quartile of the reference TMB value. In a specific embodiment, the quartile boundaries are 0 < 100 genetic alterations; 100 to 243 genetic alterations; and > 243 genetic alterations. Once ranked, it should be understood that subjects with evaluable TMB data can be divided into any number of groups, for example, quartiles, quintiles, etc.

[0088] In some embodiments, “high TMB” refers to a TMB of at least about 20 mutations / tumors, at least about 25 mutations / tumors, at least about 30 mutations / tumors, at least about 35 mutations / tumors, at least about 40 mutations / tumors, at least about 45 mutations / tumors, at least about 50 mutations / tumors, at least about 55 mutations / tumors, at least about 60 mutations / tumors, at least about 65 mutations / tumors, at least about 70 mutations / tumors, at least about 75 mutations / tumors, at least about 80 mutations / tumors, at least about 85 mutations / tumors, at least about 90 mutations / tumors, at least about 95 mutations / tumors, or at least about 100 mutations / tumors. In some embodiments, “high TMB” refers to a TMB of at least about 105 mutations / tumors, at least about 110 mutations / tumors, at least about 115 mutations / tumors, at least about 120 mutations / tumors, at least about 125 mutations / tumors, at least about 130 mutations / tumors, at least about 135 mutations / tumors, at least about 140 mutations / tumors, at least about 145 mutations / tumors, at least about 150 mutations / tumors, at least about 175 mutations / tumors, or at least about 200 mutations / tumors. In certain embodiments, a tumor having a high TMB has at least about 100 mutations / tumors.

[0089] "High TMB" may also refer to the number of mutations per megabase of a sequenced tumor genome as measured, for example, by a mutation test, for example, the FoundationOne® CDX™ test. In one embodiment, high TMB refers to at least about 9, at least about 10, at least about 11, at least 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 FoundationOne® CDX™ test. In a specific embodiment, "high TMB" refers to at least 10 mutations per megabase of the genome as measured by the FoundationOne® CDX™ test.

[0090] As used herein, the term “medium TMB” refers to the number of somatic mutations in the genome of a tumor that is or is approximately the number of normal or average somatic mutations, and the term “low TMB” refers to the number of somatic mutations in the genome of a tumor that is less than the number of normal or average somatic mutations. In certain embodiments, “high TMB” has a score of at least 243, “medium TMB” has a score of 100 to 242, and “low TMB” has a score of less than 100 (or 0 to 100). “Medium or low TMB” refers to fewer than 9 mutations per meganucleotide of the sequenced genome when measured, for example, by the FoundationOne® CDX™ assay.

[0091] The term "reference TMB value" mentioned herein may be the TMB value presented in Table 9.

[0092] In some embodiments, TMB status may be correlated with smoking status. In particular, current or former smokers often have more genetic alterations, such as missense mutations, than non-smokers.

[0093] Tumors with high TMB, for example, tumors derived from NSCLC, may also have a high neoantigen load. As used herein, the term "neoantigen" refers to a newly formed antigen that has not previously been recognized by the immune system. A neoantigen may be a protein or peptide recognized by the immune system as foreign (or non-self). Transcription of a gene within the tumor genome containing a somatic mutation produces mutated mRNA, which, upon translation, produces a mutated protein, which is then processed and transported to the ER lumen and binds to the MHC Class I complex to facilitate T-cell recognition of the neoantigen. Neoantigen recognition can promote T-cell activation, clonal expansion, and differentiation into effector and memory T-cells. Neoantigen load may correlate with TMB. In some embodiments, TMB is evaluated as a surrogate for measuring tumor neoantigen load. The TMB status of a tumor, e.g., a tumor derived from NSCLC, may be used alone or in combination with other factors as a factor in determining whether a patient is likely to benefit from a specific anticancer agent or a specific type of treatment or regimen, e.g., a combination therapy including (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. In one embodiment, a high TMB status (or high TMB) indicates an enhanced potential to benefit from immuno-oncology and may therefore be used to identify patients who are more likely to benefit from a combination therapy including (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody. Similarly, tumors with a high tumor neoantigen load and high TMB are more likely to be immunogenic than tumors with a low neoantigen load and low TMB. Additionally, high-neoantigen / high-TMB tumors are more likely to be recognized as non-self by the immune system, thereby triggering an immune-mediated antitumor response.In one embodiment, a high TMB status and a high neoantigen load represent an enhanced potential to benefit from a combination therapy, for example, immuno-oncology, comprising (1) an induction phase including administering chemotherapy to a subject for a period shorter than the standard duration for that chemotherapy; and (2) a post-induction phase including administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject after (1). As used herein, the term “benefit from therapy” refers to one or more improvements in overall survival, progression-free survival, partial response, complete response, and overall response rate, and may also include a reduction in tumor growth or size, a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or the prevention of damage or disability due to disease suffering.

[0094] Other factors, such as environmental factors, may be associated with TMB status. For example, the smoking status of patients with NSCLC is correlated with the distribution of TMB, and thus current and former smokers had higher median TMBs compared to non-smoking patients. See [Peters et al., AACR, April 1-5, 2017, Washington, DC]. The presence of driver mutations within NSCLC tumors was associated with younger female gender and non-smoking status. See [Singal et al., ASCO, June 1-5, 2017; Chicago, IL]. A tendency was observed for the presence of driver mutations, such as EGFR, ALK, or KRAS, to be associated with lower TMB (P = 0.06). See [Davis et al., AACR, April 1-5, 2017, Washington, DC].

[0095] As used herein, the term "somatic mutation" refers to acquired changes in DNA that occur after fertilization. Somatic mutations can occur in any of the body's cells, excluding germ cells (sperm and egg), and are therefore not transmitted to children. These changes can cause cancer or other diseases, but not always. The term "germ mutation" refers to genetic changes in the body's germ cells (egg or sperm) that are incorporated into the DNA of all cells within the offspring's body. Germ mutations are transmitted from parents to offspring. They are also referred to as "hereditary mutations." In the analysis of TMB, germ mutations are considered the "baseline" and are subtracted from the number of mutations found in a tumor biopsy to determine the TMB within the tumor. Because germ mutations are found in all cells within the body, their presence can be determined through sample collections that are less invasive than a tumor biopsy, such as blood or saliva. Germ mutations can increase the risk of developing certain cancers and may play a specific role in the response to chemotherapy.

[0096] When referring to TMB status, the terms “measuring,” “measured,” or “measuring” mean determining a measurable amount of somatic mutations in a biological sample of the subject. It will be recognized that the measurement may be performed by sequencing nucleic acids, e.g., cDNA, mRNA, exoRNA, ctDNA, and cfDNA, in the sample. The measurement is performed on the subject’s sample and / or reference sample or samples, and may correspond, for example, to newly detected or to a previous determination. The measurement may be performed using, as known to those skilled in the art, e.g., PCR methods, qPCR methods, Sanger sequencing methods, genome profiling methods (including comprehensive gene panels), exome sequencing methods, genome sequencing methods, and / or any other method disclosed herein. In some embodiments, the measurement identifies genomic alterations within the sequenced nucleic acids. Genome (or gene) profiling methods may involve, for example, a panel of a predetermined set of 150 to 500 genes, and in some cases, genomic alterations evaluated in the panel of genes correlate with the total somatic mutations evaluated. When referring to sequencing, the term “gene” as used herein includes DNA coding regions (e.g., exons), DNA non-coding regions associated with the coding regions (e.g., introns and promoters), and mRNA transcripts.

[0097] As used herein, the term “genome alteration” refers to a change (or mutation) in the nucleotide sequence of the genome of a tumor, such change being non-sense mutations that are not present in the wiring nucleotide sequence and include, but are not limited to, base pair substitutions, base pair insertions, base pair deletions, copy number changes (CNAs), gene rearrangements, and any combination thereof in some embodiments. In certain embodiments, the genome alteration measured in a biological sample is a missense mutation.

[0098] As used herein, the terms "whole genome sequencing" or "WGS" refer to a method of sequencing the entire genome. As used herein, the terms "whole exome sequencing" or "WES" refer to a method of sequencing all protein-coding regions (exons) of the genome.

[0099] As used herein, “cancer gene panel,” “hereditary cancer panel,” “comprehensive cancer panel,” or “polygene cancer panel” refers to a method for sequencing a subset of targeted cancer genes, including coding regions, introns, promoters, and / or mRNA transcripts. In some embodiments, the CGP comprises 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 targeted cancer genes.

[0100] The terms “genome profiling test,” “comprehensive genome profiling,” or “CGP” refer to a test that analyzes a panel of genes and selects introns for in vitro diagnosis. A CGP is a combination of NGS and targeted bioinformatics analysis for screening mutations within known clinically relevant oncogenes. These methods can be used to capture missed mutations by testing for “hotspots” (e.g., BRCA1 / BRCA2 mutations or microsatellite markers). In some embodiments, a CGP further includes one or more mRNA transcripts, non-coding RNAs, and / or promoter regions. In one embodiment, the genes within the panel are cancer-related genes. In another embodiment, the genome profiling test is a Foundation One® test.

[0101] The term "harmonization" refers to a study conducted to determine the comparability between two or more measurements and / or diagnostic tests. Harmonization studies provide a systematic approach to addressing methods for comparing diagnostic tests with one another, 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 for comparison with others. Consistent evaluations are often used in harmony studies.

[0102] As used herein, the term "concordance" refers to the degree of agreement between two types of measurements and / or diagnostic tests. Concordance may be established using both qualitative and quantitative methods. The quantitative methods used to assess concordance differ based on the type of measurement. Specific measurements may be expressed as 1) categorical / dichotomous variables or 2) continuous variables. For "categorical / dichotomous variables" (e.g., TMB above or below the cut-off), percentage concordance, such as Total Percentage Concordance (OPA), Positive Percentage Concordance (PPA), or Negative Percentage Concordance (NPA), may be used to assess concordance. For "continuous variables" (e.g., TMB by WES), Spearman rank correlation or Pearson correlation coefficient (r) is used to assess concordance across the spectrum of values, taking a value of -1 ≤ r ≤ +1 (Note: r = +1 or -1 implies that the respective variables are perfectly correlated). The term “analytical agreement” refers to the degree of agreement in the performance of two assays or diagnostic tests to support clinical use (e.g., identification of biomarkers, types of genomic alterations and genomic signatures, and evaluation of test reproducibility). The term “clinical agreement” refers to the degree of agreement in how correlated two assays or diagnostic tests are with clinical outcomes.

[0103] The term "microsatellite instability" or "MSI" refers to changes occurring in the DNA of specific cells (e.g., tumor cells) where the number of repeats of microsatellites (short, repeating DNA sequences) differs from the number of repeats present in the DNA when inherited. MSI can be high microsatellite instability (MSI-H) or low microsatellite instability (MSI-L). Microsatellites are short tandem DNA repeat sequences of 1 to 6 bases. They are prone to DNA replication errors, which are repaired by mismatch repair (MMR). Therefore, microsatellites are excellent indicators of genomic instability, particularly mismatch repair deficiency (dMMR). MSI is typically diagnosed by screening five types of microsatellite markers (BAT-25, BAT-26, NR21, NR24, and NR27). MSI-H indicates the presence of at least two unstable markers (or ≥30% of markers when a larger panel is used) among the five analyzed microsatellite markers. MSI-L indicates the instability of one MSI marker (or 10%–30% of markers in a larger panel). MSS indicates the absence of unstable microsatellite markers.

[0104] As used herein, the term “biological sample” refers to biological material isolated from a subject. The biological sample may contain any biological material suitable for determining TMB by, for example, sequencing nucleic acids within a tumor (or circulating tumor cells) and identifying genomic alterations within the sequenced nucleic acids. The biological sample may be any suitable biological tissue or fluid, e.g., tumor tissue, blood, plasma, and serum. In one embodiment, the sample is a tumor tissue biopsy, e.g., formalin-fixed paraffin-embedded (FFPE) tumor tissue or fresh-frozen tumor tissue. In another embodiment, the biological sample is a liquid biopsy comprising one or more of blood, serum, plasma, circulating tumor cells, exoRNA, ctDNA, and cfDNA, in some embodiments.

[0105] As used herein, the terms “about once a week,” “about once every two weeks,” or any other similar dosing interval term refer to an approximate number of times. “about once a week” may include every 7 ± 1 day, i.e., every 6 to 8 days. “about once every two weeks” may include every 14 ± 3 days, i.e., every 11 to 17 days. Similar approximations apply, for example, once every three weeks, once every four weeks, once every five weeks, once every six weeks, and once every twelve weeks. In some embodiments, a dosing interval of once every six weeks or once every twelve weeks means that a first dose may be administered on any day of the first week, and then the next dose may be administered on any day of the sixth week or the twelfth week, respectively. In another embodiment, an administration interval of about 6 weeks or about 12 weeks means that the first dose is administered on a specific day of the first week (e.g., Monday), and then the next dose is administered on the same day of the sixth or twelfth week (i.e., Monday), respectively.

[0106] Alternative usage (e.g., "or") should be understood to mean one, both, or any combination thereof of the alternatives. The singular form used herein should be understood to refer to "one or more" of any mentioned or enumerated components.

[0107] The terms “about” or “essentially containing” refer to a value or composition within a tolerance range for a specific value or composition as determined by a person skilled in the art, which will be partly dependent on the method by which the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, “about” or “essentially containing” may mean being within 1 or greater than 1 standard deviation according to practice in the art. Alternatively, “about” or “essentially containing” may mean a range of up to 10%. Additionally, particularly in relation to biological systems or processes, the term may mean up to 10 times or up to 5 times the value. Where a specific value or composition is provided in the application and claims, unless otherwise noted, the meaning of “about” or “essentially containing” should be assumed to be within a tolerance range for such specific value or composition.

[0108] Any concentration range, percentage range, ratio range, or integer range described herein shall be understood to include any integer value within the mentioned range, and, where appropriate, fractions thereof (e.g., 1 / 10 and 1 / 100 of an integer), unless otherwise indicated.

[0109] Various aspects of the present disclosure are further described in detail in the following subsections.

[0110] Method of the present disclosure

[0111] Certain aspects of the present disclosure relate to a method for treating a subject suffering from a tumor, comprising administering a combination therapy comprising: (1) an induction step comprising administering chemotherapy to a subject suffering from a tumor for a period shorter than the standard duration for said chemotherapy; and (2) a post-induction step comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject after (1). Some aspects of the present disclosure relate to a method for treating a tumor in a subject suffering from a tumor, comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to a subject suffering from a tumor, wherein, prior to administering the anti-PD-1 antibody or the anti-PD-L1 antibody, an induction step comprising administering chemotherapy to the subject for a period shorter than the standard duration for said chemotherapy is present. In some embodiments, the tumor originates from NSCLC.

[0112] Induction stage

[0113] In certain embodiments, the induction phase involves administering chemotherapy for a period shorter than the standard duration for that chemotherapy. In some embodiments, the induction phase involves administering a modified standard care regimen, wherein the modified standard care regimen is administered for a shorter duration than the unmodified standard care regimen. In some embodiments, chemotherapy is administered approximately once every 2 weeks, approximately once every 3 weeks, approximately once every 4 weeks, approximately once every 5 weeks, or approximately once every 6 weeks. In some embodiments, chemotherapy 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, chemotherapy is administered for 5 cycles, 4 cycles, 3 cycles, 2 cycles, or 1 cycle. In certain embodiments, chemotherapy is administered for 2 cycles, wherein each cycle involves administering chemotherapy approximately once every 3 weeks.

[0114] Standard management regimens for different types of cancer are widely 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, publishes the NCCN Guidelines for Clinical Management of Oncology (NCCN GUIDELINES®), which provide detailed and up-to-date information on standard management treatments for a wide variety of cancers (see [NCCN GUIDELINES® (2018)] (the full text of which is incorporated herein by reference), available at www.nccn.org / professionals / physician_gls / default.aspx (last accessed October 22, 2018).

[0115] For example, the NCCN guidelines for the treatment of NSCLC using chemotherapy include, but are not limited to, treatments selected from the following: (i) cisplatin 75 mg / m² 2 Day 1 plus pemetrexed 500 mg / m² 2 Day 1, for non-squamous cells every 21 days during 4 cycles; (ii) carboplatin AUC 6 Day 1, paclitaxel 200 mg / m² 2 Day 1, every 21 days for 4 cycles; and (iii) carboplatin AUC 5 Day 1, pemetrexed 500 mg / m² 2 Day 1, for non-squamous cells every 21 days for 4 cycles. Refer to [NCCN Guidelines Version 6.2018 Non-Small Cell Lung Cancer]. Other standard management chemotherapy regimens include (iv) cisplatin 50 mg / m² 2 Day 1 and Day 8 and vinorelbine 25 mg / m² 2 Day 1, Day 8, Day 15, Day 22, every 28 days during 4 cycles; (v) cisplatin 100 mg / m² 2 Day 1 and vinorelbine 30 mg / m² 2 Day 1, Day 8, Day 15, Day 22, every 28 days during 4 cycles; (vi) cisplatin 75-80 mg / m² 2Day 1 and vinorelbine 25-30 mg / m² 2 Days 1 and 8, every 21 days during 4 cycles; (vii) cisplatin 100 mg / m² 2 Day 1 and etoposide 100 mg / m² 2 Days 1–3, every 28 days for 4 cycles; (viii) cisplatin 75 mg / m² 2 Day 1 and gemcitabine 1250 mg / m² 2 Days 1 and 8, every 21 days during 4 cycles; (ix) cisplatin 75 mg / m² 2 Day 1 and docetaxel 75 mg / m² 2 Day 1, every 21 days for 4 cycles; and (x) carboplatin AUC 5 Day 1, gemcitabine 1000 mg / m² 2 Day 1 and Day 8, and every 21 days during 4 cycles. Accordingly, in an embodiment where the tumor is derived from NSCLC, e.g., stage IV NSCLC, the induction step comprises administering chemotherapy, e.g., modified standard management chemotherapy, for a period shorter than the standard duration for that chemotherapy, e.g., 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 2 cycles or less.

[0116] The chemotherapy administered during the induction phase may include any chemotherapy drugs or combinations known in the art. In certain embodiments, the chemotherapy includes standard management therapy, e.g., standard management chemotherapy. See [NCCN GUIDELINES® (2018)]. In certain embodiments, the specific chemotherapy administered depends on the histology of the tumor. For example, in some embodiments, the chemotherapy administered to treat squamous NSCLC is different from the chemotherapy administered to treat non-squamous NSCLC.

[0117] In some embodiments, the chemotherapy comprises an alkylating agent, and an antimetabolite, an antimicrotubule agent, a topoisomerase inhibitor, a cytotoxic antibiotic, or any combination thereof. In certain embodiments, the chemotherapy comprises platinum-based chemotherapy. In some embodiments, the platinum-based chemotherapy comprises cisplatin, oxaliplatin, carboplatin, nedaplatin, trilatine tetranitrate, phenantriplatin, picoplatin, satraplatin, or any combination thereof.

[0118] In certain embodiments, the chemotherapy includes platinum-based chemotherapy and a second agent. In some embodiments, the chemotherapy includes platinum-based chemotherapy and paclitaxel. In other embodiments, the chemotherapy includes platinum-based chemotherapy and pemetrexed. In some embodiments, the chemotherapy includes carboplatin and paclitaxel. In some embodiments, the chemotherapy includes carboplatin and pemetrexed. In some embodiments, the chemotherapy includes cisplatin and pemetrexed. In some embodiments, the chemotherapy includes cisplatin and paclitaxel.

[0119] In some embodiments, the induction step comprises administering chemotherapy, wherein the chemotherapy is carboplatin AUC 6 and paclitaxel 200 mg / m² on the first day of each 3-week cycle. 2 ...includes. In another embodiment, the induction step comprises administering chemotherapy, wherein the chemotherapy is carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m² on the first day of each 3-week cycle. 2 ...includes. In some embodiments, the induction step comprises administering chemotherapy, wherein the chemotherapy is carboplatin AUC 5 and pemetrexed 500 mg / m² on the first day of each 3-week cycle. 2...includes. In some embodiments, the induction step comprises administering chemotherapy, wherein the chemotherapy is carboplatin AUC 6 and pemetrexed 500 mg / m² on the first day of each 3-week cycle. 2 ...includes. In another embodiment, the induction step comprises administering chemotherapy, wherein the chemotherapy is cisplatin 75 mg / m² on the first day of each 3-week cycle. 2 and pemetrexed 500 mg / m² 2 Includes

[0120] In some embodiments, the induction step further comprises administering (i) an anti-PD-1 antibody or an anti-PD-L1 antibody and (ii) an anti-CTLA-4 antibody in addition to chemotherapy. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody is administered on the same day as chemotherapy. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody is administered on a different day from chemotherapy. In certain embodiments, (i) an anti-PD-1 antibody or an anti-PD-L1 antibody and at least one dose of (ii) an anti-CTLA-4 antibody are administered on the same day as chemotherapy.

[0121] In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody is administered at a body weight-based dose during the induction phase. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2, 3, or 4 weeks at a dose ranging from about 0.1 mg / kg to about 10.0 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 2 or 3 weeks at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 2 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 3 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 4 mg / kg body weight. In other embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 5 mg / kg body weight. In other embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 10 mg / kg body weight.

[0122] In some embodiments, the anti-PD-L1 antibody is administered at a body weight-based dose during the induction phase. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2, 3, or 4 weeks at a dose ranging from about 0.1 mg / kg to about 15.0 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 or 3 weeks at a dose of about 3 mg / kg or about 5 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 2 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 3 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 4 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 5 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 6 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 7 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 8 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 10 mg / kg body weight.

[0123] In some embodiments, the anti-CTLA-4 antibody is administered at a body weight-based dose during the induction phase. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight.

[0124] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody is administered at a uniform dose during the induction phase. In certain embodiments, both the anti-PD-1 antibody (or the anti-PD-L1 antibody) and the anti-CTLA-4 antibody are administered at a uniform dose. In other embodiments, the anti-PD-1 antibody (or the anti-PD-L1 antibody) is administered at a uniform dose, and the anti-CTLA-4 antibody is administered at a body weight-based dose. In yet another embodiment, the anti-PD-1 antibody (or the anti-PD-L1 antibody) is administered at a body weight-based dose, and the anti-CTLA-4 antibody is administered at a uniform dose.

[0125] In some embodiments, the anti-PD-1 antibody is administered during the induction phase at a uniform 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. In some embodiments, the anti-PD-1 antibody is administered once every approximately 1, 2, 3, 4, 5, or 6 weeks at a uniform dose. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 360 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of approximately 240 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of approximately 480 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of approximately 720 mg.

[0126] In some embodiments, the anti-PD-1 antibody is administered once every three weeks at a uniform dose of about 200 mg. In some embodiments, the anti-PD-1 antibody is administered once every six weeks at a uniform dose of about 400 mg. In some embodiments, the anti-PD-1 antibody is administered once every four weeks at a uniform dose of about 300 mg. In some embodiments, the anti-PD-1 antibody is administered about once a month at a uniform dose of about 300 mg. In some embodiments, the anti-PD-1 antibody is administered once every two months at a uniform dose of about 400 mg.

[0127] In some embodiments, the anti-PD-L1 antibody is administered during the induction phase at a uniform 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 once every approximately 1, 2, 3, or 4 weeks at a uniform dose. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1200 mg. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1000 mg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1100 mg. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1500 mg.

[0128] In some embodiments, the anti-CTLA-4 antibody is administered during the induction phase at a uniform 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. In some embodiments, the anti-PD-1 antibody is administered once every approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks at a uniform dose. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of approximately 80 mg.

[0129] Post-induction stage

[0130] The post-induction phase begins immediately after the induction phase. In some embodiments, the period between the first administration of the post-induction phase and the last administration of the induction phase is about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days (2 weeks), 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days (1 month), 31 days (1 month), 5 weeks, 6 weeks, 7 weeks, 8 weeks, 2 months, or 3 months or less. In some embodiments, the first dose of the post-induction phase is administered about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days (2 weeks), 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 days (1 month), 31 days (1 month), 5, 6, 7, 8 weeks, 2 months, or less than 3 months after the last dose administered during the induction phase. In certain embodiments, the first dose of the post-induction phase is administered about 3 weeks after the last dose of the induction phase. In a specific embodiment, the first dose of the post-induction phase is administered about 6 weeks after the last dose of the induction phase.

[0131] In certain embodiments, the post-induction step comprises administering immunotherapy without chemotherapy. In some embodiments, the post-induction step comprises administering an anti-PD-1 antibody or an anti-PD-L1 antibody. In certain embodiments, the post-induction step further comprises administering an anti-CTLA-4 antibody.

[0132] In some embodiments, an anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at the post-induction stage at the same dose as the anti-PD-1 antibody (or anti-PD-L1 antibody) administered at the induction stage. In some embodiments, an anti-CTLA-4 antibody is administered at the post-induction stage at the same dose as the anti-CTLA-4 antibody administered at the induction stage. In some embodiments, an anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at the post-induction stage at a different dose than the anti-PD-1 antibody (or anti-PD-L1 antibody) administered at the induction stage. In some embodiments, an anti-CTLA-4 antibody is administered at the post-induction stage at a different dose than the anti-CTLA-4 antibody administered at the induction stage.

[0133] In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody is administered at a body weight-based dose during the post-induction phase. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2, 3, or 4 weeks at a dose ranging from about 0.1 mg / kg to about 10.0 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 2 or 3 weeks at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 2 mg / kg body weight. In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 3 mg / kg body weight. In some embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 4 mg / kg body weight. In other embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 5 mg / kg body weight. In other embodiments, the anti-PD-1 antibody is administered once every three weeks at a dose of about 10 mg / kg body weight.

[0134] In some embodiments, the anti-PD-L1 antibody is administered at a body weight-based dose during the post-induction phase. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2, 3, or 4 weeks at a dose ranging from about 0.1 mg / kg to about 15.0 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 or 3 weeks at a dose of about 3 mg / kg or about 5 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 2 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 3 mg / kg body weight. In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 4 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 5 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 6 mg / kg body weight. In some embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 7 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 8 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered once every two weeks at a dose of about 10 mg / kg body weight.

[0135] In some embodiments, the anti-CTLA-4 antibody is administered at a body weight-based dose during the post-induction phase. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg body weight.

[0136] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody is administered at a uniform dose during the post-induction phase. In certain embodiments, both the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody are administered at a uniform dose. In other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a uniform dose, and the anti-CTLA-4 antibody is administered at a body weight-based dose. In yet another embodiment, the anti-PD-1 antibody (or anti-PD-L1 antibody) is administered at a body weight-based dose, and the anti-CTLA-4 antibody is administered at a uniform dose.

[0137] In some embodiments, the anti-PD-1 antibody is administered during the post-induction phase in a uniform 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. In some embodiments, the anti-PD-1 antibody is administered once every approximately 1, 2, 3, 4, 5, or 6 weeks at a uniform dose. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 360 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of approximately 240 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of approximately 480 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of approximately 720 mg.

[0138] In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 200 mg during the post-induction phase. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of about 400 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 300 mg. In some embodiments, the anti-PD-1 antibody is administered about once a month at a uniform dose of about 300 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 months at a uniform dose of about 400 mg.

[0139] In some embodiments, the anti-PD-L1 antibody is administered during the post-induction phase in a uniform 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 once every approximately 1, 2, 3, or 4 weeks at a uniform dose. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1200 mg. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1000 mg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1100 mg. In other embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of approximately 1500 mg.

[0140] In some embodiments, the anti-CTLA-4 antibody is administered during the post-induction phase in a uniform 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. In some embodiments, the anti-PD-1 antibody is administered once every approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks at a uniform dose. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of approximately 80 mg.

[0141] A specific embodiment of the present disclosure comprises (1) carboplatin AUC 6 and paclitaxel 200 mg / m² administered to a subject suffering from a tumor derived from stage IV NSCLC on the 1st day of each 3-week cycle. 2The present invention relates to a method for treating a subject suffering from a tumor derived from stage IV NSCLC, comprising administering a combination therapy including: (b) a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks; and (c) an anti-CTLA-4 antibody administered once every 6 weeks for 1 cycle at a dose of about 1 mg / kg body weight; and (2) administering a combination therapy comprising a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks and an anti-CTLA-4 antibody administered once every 6 weeks at a dose of about 1 mg / kg body weight to the subject; wherein the post-induction step is administered after the induction step. In some embodiments, the induction step continues for less than 4 cycles of chemotherapy. In some embodiments, the induction step continues for less than 3 cycles of chemotherapy. In certain embodiments, the induction step continues for 2 cycles of chemotherapy.

[0142] A specific embodiment of the present disclosure comprises (1) carboplatin AUC 5 and pemetrexed 500 mg / m² administered to a subject suffering from a tumor derived from stage IV NSCLC on the 1st day of each 3-week cycle. 2The present invention relates to a method for treating a subject suffering from a tumor derived from stage IV NSCLC, comprising administering a combination therapy including: (b) a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks; and (c) an anti-CTLA-4 antibody administered once every 6 weeks for 1 cycle at a dose of about 1 mg / kg body weight; and (2) administering a combination therapy comprising a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks and an anti-CTLA-4 antibody administered once every 6 weeks at a dose of about 1 mg / kg body weight to the subject; wherein the post-induction step is administered after the induction step. In some embodiments, the induction step continues for less than 4 cycles of chemotherapy. In some embodiments, the induction step continues for less than 3 cycles of chemotherapy. In certain embodiments, the induction step continues for 2 cycles of chemotherapy.

[0143] A specific embodiment of the present disclosure comprises (1) carboplatin AUC 6 and pemetrexed 500 mg / m² administered to a subject suffering from a tumor derived from stage IV NSCLC on the 1st day of each 3-week cycle. 2The present invention relates to a method for treating a subject suffering from a tumor derived from stage IV NSCLC, comprising administering a combination therapy including: (b) a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks; and (c) an anti-CTLA-4 antibody administered once every 6 weeks for 1 cycle at a dose of about 1 mg / kg body weight; and (2) administering a combination therapy comprising a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks and an anti-CTLA-4 antibody administered once every 6 weeks at a dose of about 1 mg / kg body weight to the subject; wherein the post-induction step is administered after the induction step. In some embodiments, the induction step continues for less than 4 cycles of chemotherapy. In some embodiments, the induction step continues for less than 3 cycles of chemotherapy. In certain embodiments, the induction step continues for 2 cycles of chemotherapy.

[0144] A specific embodiment of the present disclosure comprises (1) cisplatin 75 mg / m² administered to a subject suffering from a tumor derived from stage IV NSCLC on the 1st day of each 3-week cycle. 2 and pemetrexed 500 mg / m² 2The present invention relates to a method for treating a subject suffering from a tumor derived from stage IV NSCLC, comprising administering a combination therapy including: (b) a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks; and (c) an anti-CTLA-4 antibody administered once every 6 weeks for 1 cycle at a dose of about 1 mg / kg body weight; and (2) administering a combination therapy comprising a uniform dose of about 360 mg of an anti-PD-1 antibody administered once every 3 weeks and an anti-CTLA-4 antibody administered once every 6 weeks at a dose of about 1 mg / kg body weight to the subject; wherein the post-induction step is administered after the induction step. In some embodiments, the induction step continues for less than 4 cycles of chemotherapy. In some embodiments, the induction step continues for less than 3 cycles of chemotherapy. In certain embodiments, the induction step continues for 2 cycles of chemotherapy.

[0145] In some embodiments, the subject exhibits progression-free survival for 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 some embodiments, the subject exhibits a total 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 some embodiments, the object exhibits an objective response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0146] In some aspects, the method of the present disclosure, e.g., 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 / m² 2 or (ii) cisplatin 75 mg / m² 2 Plus Pemetrexed 500 mg / m² 2 The combination of ) demonstrates superior overall survival compared to random maintenance therapy followed by chemotherapy alone for up to 4 cycles.

[0147] In some aspects, the method of the present disclosure, e.g., 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 / m² 2 or (ii) cisplatin 75 mg / m² 2 Plus Pemetrexed 500 mg / m² 2 The combination of ) exhibits a safety profile equivalent to the known safety profiles of immunotherapy (nivolumab and / or ipilimumab) and chemotherapy components.

[0148] Anti-PD-1 antibodies useful for the present disclosure

[0149] Anti-PD-1 antibodies known in the art may be used in the compositions and methods described herein. Various human monoclonal antibodies that specifically bind to PD-1 with high affinity are disclosed in U.S. Patent No. 8,008,449. The human anti-PD-1 antibody disclosed in U.S. Patent No. 8,008,449 has been demonstrated to exhibit one or more of the following features: (a) 1 x 10⁶ when determined by surface plasmon resonance using a Biacore biosensor system -7 K less than or equal to M D(a) binds to human PD-1; (b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) increases T-cell proliferation in a mixed lymphocyte response (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 monkey 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 / or (j) inhibits tumor cell growth in vivo. Anti-PD-1 antibodies available in the present disclosure comprise monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, or at least five, of the above features.

[0150] Other anti-PD-1 monoclonal antibodies are, for example, U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509, U.S. Publication No. 2016 / 0272708, and PCT Publications WO 2012 / 145493, WO 2008 / 156712, WO 2015 / 112900, WO 2012 / 145493, WO 2015 / 112800, WO 2014 / 206107, WO 2015 / 35606, WO 2015 / 085847, WO 2014 / 179664, WO 2017 / 020291, WO 2017 / 020858, WO 2016 / 197367, WO 2017 / 024515, WO 2017 / 025051, WO 2017 / 123557, WO 2016 / 106159, WO 2014 / 194302, WO 2017 / 040790, WO 2017 / 133540, WO 2017 / 132827, WO 2017 / 024465, WO 2017 / 025016, WO 2017 / 106061, WO 2017 / 19846, WO 2017 / 024465, WO 2017 / 025016, WO 2017 / 132825, and WO 2017 / 133540 (each of these having its text attached herein It was listed in (included as a reference).

[0151] In some embodiments, the anti-PD-1 antibody is nivolumab (also known as OPDIVO®, 5C4, BMS-936558, MDX-1106 and ONO-4538), pembrolizumab (Merck; also known as KEYTRUDA®, lambrolizumab and MK-3475; see WO2008 / 156712), PDR001 (Novartis; also known as spartalizumab; see WO 2015 / 112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO 2012 / 145493), cemiplimab (Regeneron; also known as REGN-2810; see WO 2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; see reference [Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)]), BGB-A317 ("Tislelizumab"; Beigene; see WO 2015 / 35606 and US 2015 / 0079109), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210; WO 2015 / 085847; see reference [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; literature [Si-Yang Liu et al., J. Hematol. Oncol.It is selected from the group consisting of [see 10:136 (2017)]), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO 2014 / 194302), AGEN2034 (Agenus; see WO 2017 / 040790), MGA012 (Macrogenics, see WO 2017 / 19846), IBI308 (Innovent; see WO 2017 / 024465, WO 2017 / 025016, WO 2017 / 132825 and WO 2017 / 133540), and BCD-100 (Biocad).

[0152] In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that blocks the down-regulation of anti-tumor T-cell function by selectively blocking interactions with PD-1 ligands (PD-L1 and PD-L2) (U.S. Patent No. 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0153] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody directed against the human cell surface receptor PD-1 (programmed death-1 or programmed cell death-1). Pembrolizumab is described, for example, in U.S. Patents Nos. 8,354,509 and 8,900,587.

[0154] The anti-PD-1 antibodies available for use in the disclosed compositions and methods also comprise isolated antibodies that specifically bind to human PD-1 and cross-compete with any anti-PD-1 antibody disclosed herein, e.g., nivolumab, for binding to human PD-1 (see, e.g., U.S. Patents Nos. 8,008,449 and 8,779,105; WO 2013 / 173223). 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 the antibody to cross-compete for binding to an antigen indicates that these monoclonal antibodies bind to the same epitope region of the antigen and stereochemically interfere with the binding of other cross-competing antibodies to such a specific epitope region. These cross-competing antibodies are expected to have functional characteristics very similar to the reference antibody, e.g., nivolumab, in their binding to the same epitope region of PD-1. Cross-competing antibodies can be easily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays, such as Viacor assays, ELISA assays, or flow cytometry (see, for example, WO 2013 / 173223).

[0155] In certain embodiments, the antibody that cross-competes with the human PD-1 antibody, nivolumab, for binding to human PD-1, or binds to the same epitope region, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, engineered antibodies, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies may be prepared and isolated by methods widely known in the art.

[0156] The anti-PD-1 antibody usable in the compositions and methods of the present disclosure also comprises the antigen-binding portion of said antibody. It has been sufficiently demonstrated that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody.

[0157] An anti-PD-1 antibody suitable for use in the disclosed compositions and methods is an antibody that binds to PD-1 with high specificity and affinity, blocks the binding of PD-L1 and / or PD-L2, and inhibits the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, the anti-PD-1 "antibody" comprises an antigen-binding portion or fragment that binds to the PD-1 receptor, inhibits ligand binding, and exhibits functional characteristics similar to those of a whole antibody in upregulating the immune system. In certain embodiments, the anti-PD-1 antibody or its antigen-binding portion cross-competes with nivolumab for binding to human PD-1.

[0158] In some embodiments, the anti-PD-1 antibody is administered once every 2, 3, 4, 5, 6, 7, or 8 weeks at a dose ranging from 0.1 mg / kg to 20.0 mg / kg body weight, for example, once every 2, 3, or 4 weeks at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight. In other embodiments, the anti-PD-1 antibody is administered once every 2 weeks at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or 10 mg / kg body weight. In another embodiment, the anti-PD-1 antibody is administered once every 3 weeks at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or 10 mg / kg body weight. In one embodiment, the anti-PD-1 antibody is administered about once every 3 weeks at a dose of about 5 mg / kg body weight. In yet another embodiment, the anti-PD-1 antibody, e.g., nivolumab, is administered about once every 2 weeks at a dose of about 3 mg / kg body weight. In another embodiment, the anti-PD-1 antibody, e.g., pembrolizumab, is administered about once every 3 weeks at a dose of about 2 mg / kg body weight.

[0159] The anti-PD-1 antibody useful for the present disclosure may be administered in uniform doses. In some embodiments, the anti-PD-1 antibody is administered in uniform doses of about 100 to about 1000 mg, about 100 mg to about 900 mg, about 100 mg to about 800 mg, about 100 mg to about 700 mg, about 100 mg to about 600 mg, about 100 mg to about 500 mg, about 200 mg to about 1000 mg, about 200 mg to about 900 mg, about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, about 200 mg to about 500 mg, about 200 mg to about 480 mg, or about 240 mg to about 480 mg. In one embodiment, the anti-PD-1 antibody is in a homogeneous 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, about 1, 2, It is administered at intervals of 3, 4, 5, 6, 7, 8, 9, or 10 weeks.In another embodiment, the anti-PD-1 antibody is administered at equal doses of about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, and about 200 mg to about 500 mg at intervals of about 1, 2, 3, or 4 weeks.

[0160] In some embodiments, the anti-PD-1 antibody is administered about once every three weeks at a uniform dose of about 200 mg. In other embodiments, the anti-PD-1 antibody is administered about once every two weeks at a uniform dose of about 200 mg. In other embodiments, the anti-PD-1 antibody is administered about once every two weeks at a uniform dose of about 240 mg. In other embodiments, the anti-PD-1 antibody is administered about once every two weeks at a uniform dose of about 360 mg. In certain embodiments, the anti-PD-1 antibody is administered about once every four weeks at a uniform dose of about 480 mg.

[0161] Anti-PD-L1 antibodies useful for the present disclosure

[0162] Because anti-PD-1 and anti-PD-L1 target the same signaling pathway and have been found to exhibit similar levels of efficacy in various cancers, including renal cell carcinoma, in clinical trials (see references [Brahmer et al. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl J Med 366:2443-54; WO 2013 / 173223]), anti-PD-L1 antibodies may replace anti-PD-1 antibodies in any of the therapeutic methods disclosed herein. Anti-PD-L1 antibodies known in the art may be used in the compositions and methods of this disclosure. Examples of anti-PD-L1 antibodies useful in the compositions and methods of this disclosure include the antibody disclosed in U.S. Patent No. 9,580,507. The anti-PD-L1 human monoclonal antibody disclosed in U.S. Patent No. 9,580,507 has been demonstrated to exhibit one or more of the following features: (a) 1 x 10⁶ when determined by surface plasmon resonance using a Viacor biosensor system -7 K less than or equal to M D (a) binds to human PD-L1; (b) increases T-cell proliferation in a mixed lymphocyte response (MLR) assay; (c) increases interferon-γ production in an MLR assay; (d) increases IL-2 secretion in an MLR assay; (e) stimulates an antibody response; and (f) reverses the effects of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies available in the present disclosure comprise monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one of the above features, and in some embodiments, at least five.

[0163] In certain embodiments, the anti-PD-L1 antibody is BMS-936559 (also known as 12A4, MDX-1105; see, e.g., U.S. Patent No. 7,943,743 and WO 2013 / 173223), atezolizumab (Roche; also known as TECENTRIQ®; MPDL3280A, RG7446; see US 8,217,149; see also [Herbst et al. (2013) J Clin Oncol 31(suppl):3000]), durvalumab (AstraZeneca; also known as IMFINZI™, MEDI-4736; see WO 2011 / 066389), avelumab (Pfizer; also known as BAVENCIO®, MSB-0010718C). Selected from the group consisting of: announced; see WO 2013 / 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., WO 2017 / 034916), and CK-301 (Checkpoint Therapeutics; see Gorelik et al., AACR:Abstract 4606 (Apr 2016)].

[0164] In certain embodiments, the PD-L1 antibody is atezolizumab (Tecentriq®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0165] In certain embodiments, the PD-L1 antibody is durvalumab (Imfinzi™). Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0166] In certain embodiments, the PD-L1 antibody is avelumab (Bavencio®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0167] The anti-PD-L1 antibodies available for use in the disclosed compositions and methods also comprise isolated antibodies that specifically bind to human PD-L1 and cross-compete with any anti-PD-L1 antibodies disclosed herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1. In some embodiments, the anti-PD-L1 antibodies bind to the same epitope as any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab. The ability of the antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and stereochemically interfere with the binding of other cross-competing antibodies to such a specific epitope region. These cross-competing antibodies are expected to have functional characteristics very similar to the reference antibodies, e.g., atezolizumab and / or avelumab, in their binding to the same epitope region of PD-L1. Cross-competing antibodies can be easily identified based on their ability to cross-compete with atezolizumab and / or avelumab in standard PD-L1 binding assays, such as the Viacor assay, ELISA assay, or flow cytometry (see, e.g., WO 2013 / 173223).

[0168] In certain embodiments, an antibody that cross-competes with human PD-L1 antibodies, atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1, or binds to the same epitope region, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, engineered antibodies, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies may be prepared and isolated by methods widely known in the art.

[0169] The anti-PD-L1 antibody usable in the compositions and methods of the disclosed present disclosure also comprises the antigen-binding portion of said antibody. It has been sufficiently demonstrated that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody.

[0170] An anti-PD-L1 antibody suitable for use in the disclosed compositions and methods is an antibody that binds to PD-L1 with high specificity and affinity, blocks the binding of PD-1, and inhibits the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, the anti-PD-L1 "antibody" comprises an antigen-binding portion or fragment that binds to PD-L1 and exhibits functional characteristics similar to those of a whole antibody in inhibiting receptor binding and upregulating the immune system. In certain embodiments, the anti-PD-L1 antibody or its antigen-binding portion cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1.

[0171] An anti-PD-L1 antibody useful for the present disclosure may 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.

[0172] In some embodiments, the anti-PD-L1 antibody is administered about once every 2, 3, 4, 5, 6, 7, or 8 weeks at a dose ranging from about 0.1 mg / kg to about 20.0 mg / kg body weight, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg.

[0173] In some embodiments, the anti-PD-L1 antibody is administered about once every 3 weeks at a dose of about 15 mg / kg body weight. In other embodiments, the anti-PD-L1 antibody is administered about once every 2 weeks at a dose of about 10 mg / kg body weight.

[0174] In another embodiment, the anti-PD-L1 antibody useful for the present disclosure is a uniform dose. In some embodiments, the anti-PD-L1 antibody is administered in a uniform dose of about 200 mg to about 1600 mg, about 200 mg to about 1500 mg, about 200 mg to about 1400 mg, about 200 mg to about 1300 mg, about 200 mg to about 1200 mg, about 200 mg to about 1100 mg, about 200 mg to about 1000 mg, about 200 mg to about 900 mg, about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, about 700 mg to about 1300 mg, about 800 mg to about 1200 mg, about 700 mg to about 900 mg, or about 1100 mg to about 1300 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform 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, or at least about 1400 mg at intervals of about 1, 2, 3, or 4 weeks. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1,000 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1,100 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1,200 mg.In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1300 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1400 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1500 mg. In some embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 1200 mg about once every 3 weeks. In other embodiments, the anti-PD-L1 antibody is administered at a uniform dose of about 800 mg about once every 2 weeks.

[0175] Anti-CTLA-4 antibody

[0176] Anti-CTLA-4 antibodies known in the art may be used in the compositions and methods of the present disclosure. The anti-CTLA-4 antibodies of the present disclosure can bind to human CTLA-4 and disrupt the interaction between CTLA-4 and the human B7 receptor. Since the interaction between CTLA-4 and B7 transmits a signal that causes the inactivation of T-cells possessing the CTLA-4 receptor, disrupting this interaction effectively induces, enhances, or prolongs the activation of these T-cells, thereby inducing, enhancing, or prolonging the immune response.

[0177] A human monoclonal antibody that specifically binds to CTLA-4 with high affinity is disclosed in U.S. Patent No. 6,984,720. Other anti-CTLA-4 monoclonal antibodies are disclosed, for example, in U.S. Patent Nos. 5,977,318, 6,051,227, 6,682,736, and 7,034,121 and International Publications Nos. WO 2012 / 122444, WO 2007 / 113648, WO 2016 / 196237, and WO 2000 / 037504 (the full text of each of these is incorporated herein by reference). The anti-CTLA-4 human monoclonal antibody disclosed in U.S. Patent No. 6,984,720 has been demonstrated to exhibit one or more of the following features: (a) at least about 10 as determined by Viacore analysis 7 M-1 or about 10 9 M -1 or about 10 10 M -1 to 10 11 M -1 or an equilibrium association constant (K) in excess of that. a (b) specifically binds to human CTLA-4 with binding affinity reflected by ); (b) at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The dynamic association constant (k a ); (c) At least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The dynamic dissociation constant of (k d ); and (d) inhibiting the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present disclosure comprise monoclonal antibodies that specifically bind to human CTLA-4 and exhibit at least one, at least two, or at least three of the above features.

[0178] 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. Patent No. 6,984,720), MK-1308 (Merck), AGEN-1884 (Azenus Inc.; see WO 2016 / 196237), and tremelimumab (AstraZeneca; also known as ticilimumab, CP-675,206; see WO 2000 / 037504 and reference [Ribas, Update Cancer Ther. 2(3): 133-39 (2007)]). In certain embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0179] In a specific embodiment, the CTLA-4 antibody is ipilimumab for use in the compositions and methods disclosed herein. Ipilimumab is a fully human IgG1 monoclonal antibody that stimulates T cell activation and improves overall survival (OS) in patients with advanced melanoma by blocking the binding of CTLA-4 to its B7 ligand.

[0180] In a specific embodiment, the CTLA-4 antibody is tremelimumab.

[0181] In a specific embodiment, the CTLA-4 antibody is MK-1308.

[0182] In a specific embodiment, the CTLA-4 antibody is AGEN-1884.

[0183] The anti-CTLA-4 antibodies available for use in the disclosed compositions and methods also comprise isolated antibodies that specifically bind to human CTLA-4 and cross-compete with any anti-CTLA-4 antibodies disclosed herein, e.g., ipilimumab and / or tremelimumab, for binding to human CTLA-4. In some embodiments, the anti-CTLA-4 antibodies bind to the same epitope as any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab. The ability of the antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and stereochemically interfere with the binding of other cross-competing antibodies to such a specific epitope region. These cross-competing antibodies are expected to have functional characteristics very similar to the reference antibodies, e.g., ipilimumab and / or tremelimumab, in their binding to the same epitope region of CTLA-4. Cross-competing antibodies can be easily identified based on their ability to cross-compete with ipilimumab and / or tremelimumab in standard CTLA-4 binding assays, such as Viacor assays, ELISA assays, or flow cytometry (see, e.g., WO 2013 / 173223).

[0184] In certain embodiments, the antibody that cross-competes with the human CTLA-4 antibody, ipilimumab and / or tremelimumab for binding to human CTLA-4, or binds to the same epitope region, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric antibodies, engineered antibodies, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies may be prepared and isolated by methods widely known in the art.

[0185] The anti-CTLA-4 antibody usable in the compositions and methods of the present disclosure also comprises the antigen-binding portion of said antibody. It has been sufficiently demonstrated that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody.

[0186] An anti-CTLA-4 antibody suitable for use in the disclosed method or composition is an antibody that binds to CTLA-4 with high specificity and affinity, blocks the activity of CTLA-4, and disrupts the interaction between CTLA-4 and the human B7 receptor. In any of the compositions or methods disclosed herein, the anti-CTLA-4 "antibody" comprises an antigen-binding portion or fragment that binds to CTLA-4 and exhibits functional characteristics similar to a whole antibody in inhibiting the interaction between CTLA-4 and the human B7 receptor and upregulating the immune system. In certain embodiments, the anti-CTLA-4 antibody or its antigen-binding portion cross-competes with ipilimumab and / or tremelimumab for binding to human CTLA-4.

[0187] In some embodiments, the anti-CTLA-4 antibody or its antigen-binding portion is administered once every 2, 3, 4, 5, 6, 7, or 8 weeks at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody or its antigen-binding portion is administered once every 3, 4, 5, or 6 weeks at a dose of 1 mg / kg or 3 mg / kg body weight. In one embodiment, the anti-CTLA-4 antibody or its antigen-binding portion is administered once every 2 weeks at a dose of 3 mg / kg body weight. In another embodiment, the anti-PD-1 antibody or its antigen-binding portion is administered once every 6 weeks at a dose of 1 mg / kg body weight.

[0188] In some embodiments, the anti-CTLA-4 antibody or its antigen-binding portion is administered in a uniform dose. In some embodiments, the anti-CTLA-4 antibody is administered in a uniform dose of about 10 to about 1000 mg, about 10 mg to about 900 mg, about 10 mg to about 800 mg, about 10 mg to about 700 mg, about 10 mg to about 600 mg, about 10 mg to about 500 mg, about 100 mg to about 1000 mg, about 100 mg to about 900 mg, about 100 mg to about 800 mg, about 100 mg to about 700 mg, about 100 mg to about 100 mg, about 100 mg to about 500 mg, about 100 mg to about 480 mg, or about 240 mg to about 480 mg. In one embodiment, the anti-CTLA-4 antibody or its antigen-binding portion 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 It is administered in a uniform dose of 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 its antigen-binding portion is administered in a uniform dose about once every 1, 2, 3, 4, 5, 6, 7, or 8 weeks.

[0189] cytokines

[0190] In some embodiments, the method comprises administering a combination therapy comprising: (1) an induction step comprising administering chemotherapy to a subject for a period shorter than the standard duration for the chemotherapy; and (2) a post-induction step 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 step further comprises administering an anti-PD-1 antibody (or anti-PD-L1 antibody), an anti-CTLA-4 antibody, and a cytokine.

[0191] The cytokine may be any cytokine or a variant thereof known in the relevant art. 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 protein (MIP-1α and MIP-1β), IL-8, lymphotactin, fractalkine, IL-1, IL-4, IL-10, IL-11, IL-13, LIF, interferon-alpha, TGF-beta, and any combination thereof. In some embodiments, the cytokine is a CD122 agonist. In certain embodiments, the cytokine comprises IL-2 or a variant thereof.

[0192] In some embodiments, the cytokine comprises one or more amino acid substitutions, deletions, or insertions compared to the wild-type cytokine amino acid sequence. In some embodiments, the cytokine comprises an amino acid sequence in which 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 acids are substituted compared to the amino acid sequence of the wild-type cytokine.

[0193] In some embodiments, the cytokine is modified, for example, to increase its activity and / or half-life. In certain embodiments, the cytokine is modified by fusing a heterogeneous moiety to the cytokine. The heterogeneous moiety may be any structure including polypeptides, polymers, small molecules, nucleotides, or fragments or analogs thereof. In certain embodiments, the heterogeneous moiety comprises polypeptides. In some embodiments, the heterogeneous moiety comprises albumin or a fragment thereof, albumin-binding polypeptide (ABP), XTEN, Fc, PAS, the C-terminal peptide (CTP) of the β subunit of human chorionic gonadotropin, or any combination thereof.

[0194] In certain embodiments, the cytokine is modified through the fusion of the cytokine and the 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 intended to encompass any water-soluble poly(ethylene oxide). Unless otherwise indicated, the “PEG polymer” or polyethylene glycol is substantially all (preferably all) monomer subunits of which are ethylene oxide subunits, but the polymer may contain distinct terminal capping moiety or functional group for conjugation, for example. The PEG polymer for use in this disclosure will comprise one of two structures below, for example, depending on whether or not the terminal oxygen(s) were replaced during synthetic transformation: “-(CH2CH20) n-n " or "-(CH2CH20) n-1 CH2CH2-". As mentioned above, for PEG polymers, the variable (n) is in the range of about 3 to 4000, and the terminal groups and architecture of the entire PEG may vary.

[0195] In some embodiments, the method of the present disclosure comprises administering a combination therapy comprising: (1) an induction step comprising administering chemotherapy to a subject for a period shorter than the standard duration for the chemotherapy; and (2) a post-induction step 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 step 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 relative 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.

[0196] Combination therapy

[0197] In certain embodiments, an anti-PD-1 antibody, an anti-PD-L1 antibody, and / or an anti-CTLA-4 antibody are administered in a therapeutically effective amount. In some embodiments, the method comprises administering a therapeutically effective amount of an anti-PD-1 antibody and an anti-CTLA-4 antibody. In other embodiments, the method comprises administering a therapeutically effective amount of an anti-PD-L1 antibody and an anti-CTLA-4 antibody. Any anti-PD-1, anti-PD-L1, or anti-CTLA-4 antibody disclosed herein may be used in the method. 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 includes tremelimumab.

[0198] In some embodiments, (a) the anti-PD-1 antibody or the 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 the 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 the 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 once every approximately 2 weeks, once every approximately 3 weeks, or once every approximately 4 weeks, and the anti-CTLA-4 antibody is administered once every approximately 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.

[0199] In certain embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a dose of about 2 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 2 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 3 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 4 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 5 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a dose of about 10 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a dose of about 6 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg.

[0200] In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 240 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 300 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 300 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 360 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 8 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg.In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 480 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of about 480 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg.

[0201] In certain embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 240 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 3 weeks at a uniform dose of about 300 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 300 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 2 weeks at a uniform dose of about 360 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 8 weeks at a uniform dose of about 400 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg.In some embodiments, the anti-PD-1 antibody is administered once every approximately 4 weeks at a uniform dose of about 480 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-1 antibody is administered once every approximately 6 weeks at a uniform dose of about 480 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg.

[0202] In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 2 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 3 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 4 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 5 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 6 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 7 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 8 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 9 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a dose of about 10 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a dose of about 10 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg.In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a dose of about 15 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a dose of about 15 mg / kg, and the anti-CTLA-4 antibody is administered once every approximately 12 weeks at a dose of about 1 mg / kg.

[0203] In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a uniform dose of about 800 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1000 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1100 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1500 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a dose of about 1 mg / kg.

[0204] In certain embodiments, the anti-PD-L1 antibody is administered once every approximately 2 weeks at a uniform dose of about 800 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1000 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1100 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-L1 antibody is administered once every approximately 3 weeks at a uniform dose of about 1200 mg, and the anti-CTLA-4 antibody is administered once every approximately 6 weeks at a uniform dose of about 80 mg. In some embodiments, the anti-PD-L1 antibody is administered once every three weeks at a uniform dose of about 1500 mg, and the anti-CTLA-4 antibody is administered once every six weeks at a uniform dose of about 80 mg.

[0205] Tumor mutation burden

[0206] As a tumor grows, it accumulates somatic mutations that are not present in germinal DNA. TMB refers to the number of somatic mutations within the tumor genome and / or the number of somatic mutations per region of the tumor genome (after accounting for germinal variant DNA). The acquisition of somatic mutations and the resulting higher TMB can be influenced by distinct mechanisms, such as exposure to exogenous mutagenic agents (e.g., tobacco smoking) and DNA mismatch repair mutations (e.g., MSIs in colorectal and esophageal cancers). In solid tumors, approximately 95% of mutations are single-base substitutions (Vogelstein et al., Science (2013) 339:1546-1558). In this document, "nonsense mutation" refers to a nucleotide mutation that alters the amino acid sequence of a protein. Missense mutations and nonsense mutations can both be nonsense mutations. In this document, “missense mutation” refers to a non-synonymous point mutation in which a single nucleotide change produces a codon coding for a different amino acid. In this document, “nonsense mutation” refers to a non-synonymous point mutation in which a codon changes to an early stop codon that causes end cleavage of the resulting protein.

[0207] In one embodiment, somatic mutations may be expressed at the RNA and / or protein level to generate neoantigens (also referred to as neoepitopros). Neoantigens may influence immune-mediated antitumor responses. For example, recognition of neoantigens may promote T-cell activation, clonal expansion, and differentiation into effector and memory T-cells.

[0208] As tumors develop, early clonal mutations (or "stem mutations") may be retained by most or all tumor cells, while later mutations (or "branching mutations") may occur only in a subset of tumor cells or 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 widespread in the tumor genome than "branching" mutations, and thus can induce a large number of T cells reactive to clonal neoantigens (McGranahan et al., (2016) 351:1463-1469). In general, tumors with high TMB may also have a high neoantigen load, which can lead to high tumor immunogenicity and increased T-cell responsiveness and anti-tumor responses. Therefore, cancers with high TMB may respond well to immunotherapy, for example, treatment with anti-PD-1 antibodies or anti-PD-L1 antibodies. For example, refer to International Publication No. WO / 2018 / 183928 A1 (the full text of which is incorporated herein by reference).

[0209] Advancements in sequencing technology enable the evaluation of the genomic mutation landscape of tumors. Nucleic acids from a tumor genome (e.g., obtained from a biological sample from a subject suffering from a tumor) can be sequenced using any sequencing method known to a person skilled in the art. In one embodiment, PCR or qPCR methods, Sanger sequencing methods, or next-generation sequencing ("NGS") methods (e.g., genome profiling, exome sequencing, or genome sequencing) may be used to measure TMB. In some embodiments, the TMB status is measured using genome profiling. Genome profiling involves analyzing nucleic acids from a tumor sample, including coding and non-coding regions, which may be performed using a method that incorporates optimized nucleic acid selection, read alignment, and mutation calling. In some embodiments, gene profiling provides a next-generation sequencing (NGS)-based analysis of the tumor that can be optimized on a cancer-specific, gene-specific, and / or site-specific basis. Genome profiling can incorporate the use of multiple individually tuned alignment methods or algorithms to optimize performance in sequencing methods, particularly those relying on large-scale parallel sequencing of numerous diverse genetic events across many diverse genes. Genome profiling provides a comprehensive analysis of a subject's cancer genome of clinical-grade quality, and the results of genetic analysis can be contextualized with relevant scientific and medical knowledge to enhance the quality and efficiency of cancer therapy.

[0210] Genome profiling involves a panel of a predetermined set of genes, comprising at least 5 genes or as many as 1,000 genes, about 25 genes to about 750 genes, about 100 genes to about 800 genes, about 150 genes to about 500 genes, about 200 genes to about 400 genes, and about 250 genes to about 350 genes. In one embodiment, the genome 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 genome profile comprises at least 325 genes. In a specific embodiment, the genome profile comprises at least 315 cancer-associated genes and introns within 28 genes (FoundationOne®) or the complete DNA coding sequences of 406 genes, introns within 31 genes by rearrangement, and RNA sequences (cDNA) of 265 genes (FoundationOne® Heme). In another embodiment, the genome profile comprises 26 genes and 1,000 associated mutations (EXODX® Solid Tumor). In another embodiment, the genome profile comprises 76 genes (Guardant360). In another embodiment, the genome profile comprises 73 genes (Guardant360).In another embodiment, the genome profile includes 354 genes and introns within 28 genes for rearrangement (FoundationOne® CDX™). In a specific embodiment, the genome profile is FoundationOne® F1CDx. In another embodiment, the genome profile includes 468 genes (MSK-IMPACT™). One or more genes may be added to the genome profile because more genes are identified as being relevant to oncology.

[0211] Foundation One® Black

[0212] The Foundation One® assay is a comprehensive genomic profiling assay for solid tumors, including but not limited to solid tumors of the lung, colon, and breast, melanoma, and ovarian cancer. The Foundation One® assay uses hybrid-capture, next-generation sequencing tests to identify genomic alterations (base substitutions, insertions and deletions, copy number changes, and rearrangements) and to select genomic signatures (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 Foundation One® assay genes is provided in Tables 1A and 1B. Refer to [FOUNDATIONONE: Technical Specifications, Foundation Medicine, Inc.] (the full text of which is incorporated herein by reference), available at FoundationMedicine.com (last accessed March 16, 2018).

[0213] Table 1A: List of genes whose whole coding sequences were tested in the Foundation One® test.

[0214]

[0215]

[0216] Table 1B: List of genes tested for introns selected in the Foundation One® test.

[0217]

[0218] EXODX® Solid Tumor Test

[0219] In one embodiment, TMB is measured using the EXODX® Solid Tumor Test. The EXODX® Solid Tumor Test is an exoRNA- and cfDNA-based test that detects mutations capable of acting in cancer pathways. The EXODX® Solid Tumor Test is a plasma-based test that does not require tissue samples. The EXODX® Solid Tumor Test covers 26 genes and 1,000 mutations. Specific genes covered by the EXODX® Solid Tumor Test are presented in Table 2. Refer to [Plasma-Based Solid Tumor Mutation Panel Liquid Biopsy, Exosome Diagnostics, Inc.] available at exosomedx.com (last accessed March 16, 2018).

[0220] Table 2: Genes covered by the EXODX® solid tumor assay.

[0221]

[0222] Foundation One® Liquid Black

[0223] In one embodiment, TMB is measured using the Foundation One® Liquid assay. The Foundation One® 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 tissue sample. The Foundation One® Liquid assay covers 70 genes. Specific genes covered by the Foundation One® Liquid assay are presented in Tables 3A–3C. Refer to [FOUNDATIONONE® Liquid, Technical Specifications, Foundation Medicine], available at assets.ctfassets.net / vhribv12lmne / 3SPYAcbGdqAeMsOqMyKUog / d0eb51659e08d733bf39971e85ed940d / F1L_TechnicalInformation_MKT-0061-04.pdf (last accessed October 6, 2018).

[0224] Table 3A: Genes covered by Foundation One® Liquid Assay: Whole coding sequence.

[0225]

[0226] Table 3B: Genes covered by Foundation One® Liquid Assay: Selected exons.

[0227]

[0228] Table 3C: Genes covered by Foundation One® liquid assay: Selective rearrangement.

[0229]

[0230] Guardant 360 Black

[0231] In some embodiments, TMB status is determined using the Guardant360 test. The Guardant360 test 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™ test. The GuardantOmni™ test is a comprehensive genome profiling tool containing a 500-gene panel.

[0232] Table 4A: Guardant360 test gene.

[0233]

[0234] Table 4B: Guardant360 test indel.

[0235]

[0236] Table 4C: Guardant360 Test Amplification (CNV).

[0237]

[0238] Table 4D: Guardant360 test fusion.

[0239]

[0240] ILLUMINA® TruSight Test

[0241] In some embodiments, TMB is determined using the TrueSite Tumor 170 assay (Illumina). The TrueSite Tumor 170 assay is a next-generation sequencing assay that covers 170 genes associated with common solid tumors and analyzes DNA and RNA simultaneously. The TrueSite Tumor 170 assay evaluates fusions, splice variants, insertions / deletions, single nucleotide variants (SNVs), and amplifications. A list of TrueSite Tumor 170 assay genes is presented in Tables 5A-5C.

[0242] Table 5A: TrueSite Tumor 170 Test Gene (Amplification).

[0243]

[0244] Table 5B: TrueSite Tumor 170 Test Gene (Fusion).

[0245]

[0246] Table 5C: TrueSite Tumor 170 Test Gene (Small Variant).

[0247]

[0248] Foundation One® F1CDx Black

[0249] FoundationOne® CDX™ ("F1CDx") is a next-generation sequencing-based in vitro diagnostic device for detecting genomic signatures, including microsatellite instability (MSI) and tumor mutation burden (TMB), using DNA isolated from formalin-fixed paraffin-embedded (FFPE) tumor tissue specimens, as well as substitutions, insertions, and deletions (indels) and copy number changes (CNAs) in 324 genes and selected gene rearrangements. F1CDx is approved by the U.S. Food and Drug Administration (FDA) for several tumor indications, including NSCLC, melanoma, breast cancer, colorectal cancer, and ovarian cancer.

[0250] The F1CDx assay utilizes a single DNA extraction method from commercial FFPE biopsies or surgically resected specimens; 50–1000 ng of these specimens will undergo the construction of a whole-genome shotgun library and hybridization-based capture of all coding exons, one promoter region, and one non-coding (ncRNA) from 309 cancer-associated genes, as well as selected intron regions from 34 commonly rearranged genes (of which 21 also contain coding exons). Tables 6A and 6B provide a complete list of genes included in F1CDx. Overall, the assay detects alterations in a total of 324 genes. Using the Illumina® HiSeq 4000 platform, the hybridized-captured-selected library is sequenced to a high, uniform depth (targeted with >500X median coverage, and >99% of exons have >100X coverage). Next, sequence data is processed using a custom analysis pipeline designed to detect all classes of genomic alterations, including base substitutions, indels, copy number changes (amplification and homozygous gene deletions), and selected genomic rearrangements (e.g., gene fusions). Additionally, genomic signatures including microsatellite instability (MSI) and tumor mutation burden (TMB) are reported.

[0251] Table 6A: Genes containing the entire coding exon region included in FoundationOne® CDX™ for the detection of substitutions, insertions and deletions (indels), and copy number changes (CNA).

[0252]

[0253] Table 6B: Genes having selected intron regions for the detection of gene rearrangements (one has a 3'UTR, one has a promoter region, and one is an ncRNA gene).

[0254]

[0255] F1CDx assays identify various alterations in gene and / or intron sequences, including substitutions, insertions / deletions, and CNAs. F1CDx assays have been previously confirmed to be consistent with externally validated NGS assays and FoundationOne® (F1 LDT) assays. Refer to [FOUNDATIONONE® CDX™: Technical Information, Foundation Medicine, Inc.] (the full text of which is incorporated herein by reference), available at FoundationMedicine.com (last accessed March 16, 2018).

[0256] MSK-IMPACT™

[0257] In some embodiments, TMB status is evaluated using the MSK-IMPACT™ assay. The MSK-IMPACT™ assay uses next-generation sequencing to analyze the mutation status of 468 genes. Target genes are captured and sequenced on Illumina HISEQ™ instruments. The MSK-IMPACT™ assay is approved by the U.S. FDA for the detection of somatic mutations and microsatellite instability in solid malignant neoplasms. A complete list of the 468 genes analyzed by the MSK-IMPACT™ assay is presented in Table 7. Refer to [Evaluation of Automatic Class III Designation for MSK-IMPACT (Integrated Mutation Profiling of Actionable Cancer Targets): Decision Summary, United States Food and Drug Administration, November 15, 2017], available at accessdata.fda.gov.

[0258] Table 7: Genes analyzed by the MSK-IMPACT™ assay.

[0259]

[0260]

[0261] NEOGENOMICS® NEOTYPE™ Test

[0262] In some embodiments, TMB is determined using the NeoGenomics® NeoType™ assay. In some embodiments, TMB is determined using the NeoType™ discovery profile. In some embodiments, TMB is determined using the NeoType solid tumor profile. The NeoGenomics assay measures the number of non-synonymic DNA coding sequence changes per megabase of sequenced DNA.

[0263] ONCOMINE™ Tumor Mutation Load Test

[0264] In some embodiments, TMB is determined using the ThermoFisher Scientific® Oncomin™ Tumor Mutation Test. In some embodiments, TMB is determined using the ThermoFisher Scientific® ION TORRENT™ Oncomin™ Tumor Mutation Test. The ION TORRENT™ Oncomin™ Tumor Mutation Test is a targeted NGS test that quantifies somatic mutations to determine the tumor mutation load. The test covers 1.7 Mb of DNA.

[0265] NOVOGENE™ NOVOPM™ Test

[0266] In some embodiments, TMB is determined using the Novogen™ NovoPM™ assay. In some embodiments, TMB is determined using the Novogen™ NovoPM™ cancer panel assay. The Novogen™ NovoPM™ cancer panel assay analyzes the complete coding regions of 548 genes and the introns of 21 genes, representing approximately 1.5 Mb of DNA, and is a comprehensive NGS cancer panel related to the diagnosis and / or treatment of solid tumors according to National Comprehensive Cancer Network (NCCN) guidelines and medical literature. The assay detects genomic abnormalities such as SNVs, InDels, fusions, and copy number variants (CNVs).

[0267] Other TMB tests

[0268] In some embodiments, TMB is determined using a TMB test provided by CARIS® Life Sciences. In some embodiments, TMB is determined using the PESONALIS® ACE ImmunoID test. In some embodiments, TMB is determined using the PGDX® CANCERXOME™-R test.

[0269] In another specific embodiment, genome profiling detects all mutation types, namely single nucleotide variants, insertions / deletions (indels), copy number variations, and rearrangements, e.g., translocations, expressions, and epigenetic markers.

[0270] Comprehensive gene panels often contain predetermined genes selected based on the type of tumor to be analyzed. Thus, a genome profile used to measure TMB status can be selected based on the type of tumor the subject has. In one embodiment, the genome profile may include a specific set of genes for solid tumors. In another embodiment, the genome profile may include a specific set of genes for hematological malignancies and sarcomas.

[0271] 한 실시양태에서, 게놈 프로파일은 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 (C17orf39), 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), C11orf30 (EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1T1, TERT (프로모터만), 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, CCND2, DAXX, FGF23, GSK3B, KMT2A (MLL), NF1, POLD1, SETD2, TOP1, ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C (MLL3), NF2POLE, SF3B1, TOP2A, ARID2, CCNE1, 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, CDC73, 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, 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, BCL6, CDKN2C, ESR1, FUBP1,It includes one or more genes selected from the group consisting of 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 any combination thereof. In another embodiment, TMB analysis further includes identifying genomic alterations in one or more of ETV4, TMPRSS2, ETV5, BCR, ETV1, ETV6, and MYB.

[0272] 또 다른 실시양태에서, 게놈 프로파일은 ABL1, 12B, ABL2, ACTB, ACVR1, ACVR1B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, AMER1 (FAM123B 또는 WTX), AMER1 (FAM123B), ANKRD11, APC, APH1A, AR, ARAF, ARFRP1, ARHGAP26 (GRAF), ARID1A, ARID1B, ARID2, ARID5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AURKB, AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL10, BCL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1, BRIP1 (BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf 30 (EMSY), C11orf30, C11orf30 (EMSY), CAD, CALR, CARD11, CARM1, CASP8, CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274 (PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CDC42, 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, 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, EML4-ALK, EP300, EPAS1, EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2, ERBB3, ERBB4, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, ERF, ERG, ERRFI1, ERRFl1, ESR1, 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, FGF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGFR1, FGFR2, FGFR3, FGFR4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH1, FOXA1, FOXL2, FOXO1, FOXO3, FOXP1, FRS2, FUBP1, FYN, GABRA6, GADD45B, GATA1, GATA2, GATA3, GATA4, GATA6, GEN1, GID4 (C17orf 39), GID4 (C17orf39), GLI1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRIN2A, GRM3, GSK3B, GTSE1, H3F3A, H3F3B,H3F3C, HDAC1, HDAC4, HDAC7, Hedgehog, HER-2 / NEU; ERBB2, HGF, HIST1H1C, HIST1H1D, HIST1H1E, HIST1H2AC, HIST1H2AG, HIST1H2AL, HIST1H2AM, HIST1H2BC, HIST1H2BD, HIST1H2BJ, HIST1H2BK, HIST1H2BO, HIST1H3A, HIST1H3B, HIST1H3C, HIST1H3D, HIST1H3E, HIST1H3F, HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HIST2H3C, HIST2H3D, 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 (MYST 3), 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, MAP2K1, MAP2K1 (MEK1), MAP2K2, MAP2K2 (MEK2), MAP2K4, MAP3, MAP3K1, MAP13, MAP14, 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, MRE 11A, MRE11A, MSH2, MSH3, MSH6, MSI1, MSI2, MST1, MST1R, MTAP, MTOR, MUTYH, MYC, MYCL, MYCL (MYC L1), MYCL (MYCL1), MYCL1, MYCN, MYD88, MYO18A, MYOD1, NBN, NCOA3, NCOR1, NCOR2, NCSTN, NEGR1, NF1, NF2, NFE2L2, NFKBIA, NKX2-1, 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, TBL1XR1, TBX3, TCEB1, TCF3, TCF3 (E2A), TCF7L2, TCL1A (TCL1), TEK, TERC, TERT, TERT 프로모터, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL2, TMEM127, TMEM30A, TMPRSS2, TMSB4XP8 (TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14It comprises one or more genes selected from the group consisting of 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), WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and any combination thereof.

[0273] In another embodiment, genome profiling tests ABL1, 12B, ABL2, ACTB, ACVR1, ACVR1B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, AMER1 (FAM123B or WTX), AMER1 (FAM123B), ANKRD11, APC, APH1A, AR, ARAF, ARFRP1, ARHGAP26 (GRAF), ARID1A, ARID1B, ARID2, ARID5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AURKB, AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL10, BCL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1, BRIP1 (BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf 30 (EMSY), C11orf30, C11orf30 (EMSY), CAD, CALR, CARD11, CARM1, CASP8, CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274 (PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CDC42, 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, 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, EML4-ALK, EP300, EPAS1, EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2, ERBB3, ERBB4, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, ERF, ERG, ERRFI1, ERRFl1, ESR1, 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, FGF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGFR1, FGFR2, FGFR3, FGFR4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH1, FOXA1, FOXL2, FOXO1, FOXO3, FOXP1, FRS2, FUBP1, FYN, GABRA6, GADD45B, GATA1, GATA2, GATA3, GATA4, GATA6, GEN1, GID4 (C17orf 39), GID4 (C17orf39), GLI1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRIN2A, GRM3, GSK3B, GTSE1, H3F3A,H3F3B, H3F3C, HDAC1, HDAC4, HDAC7, Hedgehog, HER-2 / NEU; ERBB2, HGF, HIST1H1C, HIST1H1D, HIST1H1E, HIST1H2AC, HIST1H2AG, HIST1H2AL, HIST1H2AM, HIST1H2BC, HIST1H2BD, HIST1H2BJ, HIST1H2BK, HIST1H2BO, HIST1H3A, HIST1H3B, HIST1H3C, HIST1H3D, HIST1H3E, HIST1H3F, HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HIST2H3C, HIST2H3D, 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 (MYST 3), 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, MAP2K1, MAP2K1 (MEK1), MAP2K2, MAP2K2 (MEK2), MAP2K4, MAP3, MAP1, MAP13, MAP14, MAP6, MAP7,MAPK1, MAPK3, MAPKAP1, MAX, MCL1, MDC1, MDM2, MDM4, MED12, MEF2B, MEF2C, MEK1, MEN1, MERTK, MET, MGA, MIB1, MITF, MKI67, MKNK1, MLH1, MLLT3, MPL, MRE 11A, MRE11A, MSH2, MSH3, MSH6, MSI1, MSI2, MST1, MST1R, MTAP, MTOR, MUTYH, MYC, MYCL, MYCL (MYC L1), MYCL (MYCL1), MYCL1, MYCN, MYD88, MYO18A, MYOD1, NBN, NCOA3, NCOR1, NCOR2, NCSTN, NEGR1, NF1, NF2, NFE2L2, NFKBIA, NKX2-1, 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, TBL1XR1, TBX3, TCEB1, TCF3, TCF3 (E2A), TCF7L2, TCL1A (TCL1), TEK, TERC, TERT, TERT 프로모터, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL2, TMEM127, TMEM30A, TMPRSS2, TMSB4XP8 (TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14At least about 20, at least about 30, at least about 40, and at least about selected from the group consisting of 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), WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and any combination thereof It includes 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, 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.

[0274] In another embodiment, the genome profile includes one or more genes selected from the genes listed in Table 2-14.

[0275] In one embodiment, TMB status based on genome profiling is highly correlated with TMB status based on whole-exome or whole-genome sequencing. Evidence shows that the use of genome profiling assays, such as the F1CDx assay, is consistent with whole-exome and / or whole-genome sequencing assays. These data support the use of genome profiling assays as a more efficient means of measuring TMB status without compromising the prognostic quality of the TMB status.

[0276] TMB can be measured using tissue biopsy samples, or alternatively, circulating tumor DNA (ctDNA), cfDNA (cell-free DNA), and / or liquid biopsy samples. ctDNA can be used to measure TMB status based on whole-exome or whole-genome sequencing or genomic profiling using methodologies available from, for example, GRAIL, Inc.

[0277] In some embodiments, subjects are identified as suitable for the combination therapy disclosed herein based on the measurement of TMB status and the confirmation of high TMB. In some embodiments, the TMB score is calculated as the total number of non-synonymous missense mutations within the tumor when measured by whole-exome sequencing or whole-genome sequencing. In one embodiment, the high 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 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 It has a score of 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, the 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 a specific embodiment, the high TMB has a score of at least 243. In another embodiment, the high TMB has a score of at least 244. In some embodiments, the high TMB has a score of at least 245. In other embodiments, the high TMB has a score of at least 246. In other embodiments, the high TMB has a score of at least 247. In other embodiments, the high TMB has a score of at least 248. In other embodiments, the high TMB has a score of at least 249. In other embodiments, the high TMB has a score of at least 250. In other embodiments, the high TMB has any integer score between 200 and 300 or greater. In other embodiments, the high TMB has any integer score between 210 and 290 or greater. In other embodiments, the high TMB has any integer score between 220 and 280 or greater. In other embodiments, the high TMB has any integer score between 230 and 270 or greater. In other embodiments, the high TMB has any integer score between 235 and 265 or greater.

[0278] Alternatively, a high TMB may be a relative value rather than an absolute value. In some embodiments, the TMB state of the object is compared to a reference TMB value. In one embodiment, the TMB state of the object is within the highest quartile of the reference TMB value. In another embodiment, the TMB state of the object is within the upper quartile of the reference TMB value.

[0279] In some embodiments, the TMB status is expressed as the number of mutations per sample, per cell, per exome, or per DNA length (e.g., Mb). In some embodiments, if a tumor has at least about 50 mutations / tumors, at least about 55 mutations / tumors, at least about 60 mutations / tumors, at least about 65 mutations / tumors, at least about 70 mutations / tumors, at least about 75 mutations / tumors, at least about 80 mutations / tumors, at least about 85 mutations / tumors, at least about 90 mutations / tumors, at least about 95 mutations / tumors, at least about 100 mutations / tumors, at least about 105 mutations / tumors, at least about 110 mutations / tumors, at least about 115 mutations / tumors, or at least about 120 mutations / tumors, the tumor has a high TMB status. In some embodiments, if the tumor has at least about 125 mutations / tumors, at least about 150 mutations / tumors, at least about 175 mutations / tumors, at least about 200 mutations / tumors, at least about 225 mutations / tumors, at least about 250 mutations / tumors, at least about 275 mutations / tumors, at least about 300 mutations / tumors, at least about 350 mutations / tumors, at least about 400 mutations / tumors, or at least about 500 mutations / tumors, the tumor has a high TMB state. In one specific embodiment, if the tumor has at least about 100 mutations / tumors, the tumor has a high TMB state.

[0280] In some embodiments, the tumor contains at least about 5 mutations (mutations / Mb), at least about 6 mutations / Mb, at least about 7 mutations / Mb, at least about 8 mutations / Mb, at least about 9 mutations / Mb, at least about 10 mutations / Mb, at least about 11 mutations / Mb, at least about 12 mutations / Mb, at least about 13 mutations / Mb, at least about 14 mutations / Mb, at least about 15 mutations / Mb, at least about 20 mutations / Mb, at least about 25 mutations / Mb, at least about 30 mutations / Mb, at least about 35 mutations / Mb, at least about 40 mutations / Mb, at least about 45 mutations / Mb, at least about 50 mutations / Mb, at least about If the tumor has 75 mutations / Mb or at least about 100 mutations / Mb, the tumor has a high TMB state. In certain embodiments, if the tumor has at least about 5 mutations / Mb, the tumor has a high TMB state. In certain embodiments, if the tumor has at least about 10 mutations / Mb, the tumor has a high TMB state. In some embodiments, if the tumor has at least about 11 mutations / Mb, the tumor has a high TMB state. In some embodiments, if the tumor has at least about 12 mutations / Mb, the tumor has a high TMB state. In some embodiments, if the tumor has at least about 13 mutations / Mb, the tumor has a high TMB state. In some embodiments, if the tumor has at least about 14 mutations / Mb, the tumor has a high TMB state. In certain embodiments, if the tumor has at least about 15 mutations / Mb, the tumor has a high TMB state.

[0281] Because the number of mutations varies depending on the tumor type and other factors (see Q4 and Q5), the values ​​associated with "high TMB" and "low TMB" may differ across tumor types.

[0282] PD-L1 status

[0283] TMB status may be used alone or in combination with other factors as a means to predict the response of a tumor to a combination therapy comprising (a) an anti-PD-1 antibody or an 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 more likely to respond to a combination therapy comprising (a) an anti-PD-1 antibody or an 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 more likely to respond to 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 tumor has less than 1% PD-L1 expression, for example, less than 1% of tumor cells express PD-L1. In certain embodiments, the subject has a high TMB status (≥10 mutations / Mb) and a tumor PD-L1 expression level of less than 1%.

[0284] The PD-L1 status of the tumor in the subject may be measured before administering any of the compositions disclosed herein or before using any of the methods disclosed herein. PD-L1 expression may be determined by any method known in the relevant art.

[0285] To evaluate PD-L1 expression, in one embodiment, a test tissue sample may be obtained from a patient requiring therapy. In another embodiment, the evaluation of PD-L1 expression may be achieved without obtaining a test tissue sample. In some embodiments, selecting a suitable patient comprises: (i) optionally providing a test tissue sample comprising tumor cells and / or tumor-infiltrating inflammatory cells obtained from a patient having a tumor derived from NSCLC; and (ii) evaluating the proportion of cells expressing PD-L1 on the cell surface in the test tissue sample based on an assessment that the proportion of cells expressing PD-L1 on the cell surface in the test tissue sample is higher than a predetermined threshold level.

[0286] However, in any method comprising measuring PD-L1 expression in a test tissue sample, the step of providing a test tissue sample obtained from a patient should be understood as an optional step. Additionally, in certain embodiments, the "measurement" or "evaluation" step for identifying cells expressing PD-L1 on the cell surface of a test tissue sample or determining their number or proportion should be understood as being performed by a variant assay method for PD-L1 expression, for example, by performing a reverse transcription-polymerase chain reaction (RT-PCR) assay or an IHC assay. In other specific embodiments, without any variant steps, PD-L1 expression is evaluated, for example, by reviewing a test result report from a laboratory. In certain embodiments, the method step up to and including the evaluation of PD-L1 expression provides intermediate results that may be provided to a physician or other healthcare provider for use in selecting a suitable candidate 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 intermediate results is performed by a medical practitioner or a person acting under the direction of a medical practitioner. In other embodiments, these steps are performed by an independent laboratory or by an independent person, such as a laboratory technician.

[0287] In any specific embodiment of the present method, the proportion of cells expressing PD-L1 is evaluated by performing a assay to determine the presence of PD-L1 RNA. In additional 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 evaluated by performing a assay to determine the presence of PD-L1 polypeptide. In additional 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 evaluated by IHC. In other embodiments of all these methods, cell surface expression of PD-L1 is evaluated, for example, using IHC or in vivo imaging.

[0288] Imaging technology has provided important tools for cancer research and treatment. Recent advancements in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflection imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), laser-scanning confocal microscopy (LSCM), and multiphoton microscopy (MPM), foreshadow the potential for these technologies to be used much more extensively in cancer research. Some of these molecular imaging systems not only enable clinicians to identify the location of tumors within the body but also allow for the visualization of the expression and activity of specific molecules, cells, and biological processes that influence tumor behavior and / or responsiveness to therapeutic drugs (Condeelis and Weissleder, "In vivo imaging in cancer," Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Antibody specificity coupled with the sensitivity and resolution of PET makes immunoPET imaging particularly attractive for monitoring and assessing antigen expression 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 any specific embodiment of the present method, PD-L1 expression is assessed by immunoPET imaging.In any specific embodiment of the present method, the proportion of cells expressing PD-L1 in a test tissue sample is evaluated by performing a assay to determine the presence of PD-L1 polypeptide on the cell surface of the test tissue sample. In a specific embodiment, the test tissue sample is an FFPE tissue sample. In another embodiment, the presence of PD-L1 polypeptide is determined by an IHC assay. In additional embodiments, the IHC assay is performed using an automated process. In some embodiments, the IHC assay is performed using an anti-PD-L1 monoclonal antibody that binds to PD-L1 polypeptide. In certain embodiments, the anti-PD-L1 monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and any combination thereof. Refer to WO / 2013 / 173223 (the full text of which is incorporated herein by reference).

[0289] In one embodiment of the present method, an automated IHC method is used to test for the expression of PD-L1 on the cell surface of FFPE tissue specimens, for example, tissue samples taken from tumors derived from NSCLC. The presence of human PD-L1 antigen can be measured in the test tissue sample by contacting the test sample and negative control sample (e.g., normal tissue) with a monoclonal antibody that specifically binds to human PD-L1, under conditions that allow for the formation of a complex between the antibody or a portion thereof and human PD-L1. In a specific embodiment, the test and control tissue samples are FFPE samples. Subsequently, the formation of a complex is detected, wherein the 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 are used to quantify PD-L1 expression.

[0290] In a specific embodiment, the automated IHC method comprises operating an automated stainer comprising the steps of: (a) deparaffinizing and rehydrating a tissue section loaded in an automated stainer; (b) recovering an antigen using a declocking chamber heated to 110°C for 10 minutes and a pH 6 buffer; (c) setting a reagent on the automated stainer; and (d) neutralizing endogenous peroxidase in the tissue specimen; blocking non-specific protein-binding sites on the slide; incubating the slide with a primary antibody; incubating with a primary post-blocker; incubating with a Novolink polymer; adding a chromogen substrate and developing; and counterstaining with hematoxylin.

[0291] To evaluate PD-L1 expression in tumor tissue samples, pathologists examine the PD-L1 within each field of the membrane under a microscope. +The number of tumor cells is examined, the percentage of positive cells is estimated by the genus, and then averaged to obtain a final percentage. Different staining intensities are defined as 0 / negative, 1+ / weak, 2+ / medium, and 3+ / strong. Typically, percentage values ​​are first assigned to the 0 and 3+ buckets, and then the medium 1+ and 2+ intensities are considered. For highly heterogeneous tissues, the specimen is divided into multiple zones, each zone is scored individually, and then summed into a single set of percentage values. The percentages of negative and positive cells for different staining intensities are determined from each zone, and a median is provided for each zone. A final percentage value is given to the tissue for each of the following staining intensity categories: negative, 1+, 2+, and 3+. The sum of all staining intensities must be 100%. In one embodiment, the threshold number of cells that must be PD-L1 positive 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 that must be PD-L1 positive is at least about 100 cells. In some embodiments, the tumor sample must have at least 100 total tumor cells so that the tumor sample is considered a specimen that can be evaluated for PD-L1 expression.

[0292] Staining is also evaluated for tumor-infiltrating inflammatory cells, such as macrophages and lymphocytes. In most cases, macrophages serve as an internal positive control because staining is observed in a large proportion of macrophages. Although staining to a 3+ intensity is not required, the absence of macrophage staining should be considered to rule out any technical failures. Macrophages and lymphocytes are evaluated for plasma membrane staining and are recorded as positive or negative for their respective cell categories for all samples. Staining is also characterized according to external / internal tumor immune cell designations. "Internal" means that immune cells are located within the tumor tissue and / or at the boundary of the tumor region without being physically interbedded among tumor cells. "External" means that immune cells are found in the periphery associated with connective tissue or in any associated adjacent tissue, without any physical association with the tumor.

[0293] In a specific embodiment of these scoring methods, samples are scored by two pathologists working independently, and the scores are subsequently integrated. In another specific embodiment, the identification of positive and negative cells is scored using appropriate software.

[0294] Histoscore is used as a more quantitative measure of IHC data. Histoscore is calculated as follows:

[0295] Histoscore = [(% Tumor x 1 (Low Intensity)) + (% Tumor x 2 (Medium Intensity)) + (% Tumor x 3 (High Intensity)]

[0296] To determine the histoscore, a pathologist estimates the percentage of stained cells in each intensity category within the specimen. Because the expression of most biomarkers is heterogeneous, the histoscore is a more accurate representation of total expression. The final histoscore ranges from 0 (no expression) to 300 (maximum expression).

[0297] IHC, an alternative means of quantifying PD-L1 expression in test tissue samples, determines the Adjusted Inflammation Score (AIS), which is defined as the percentage of PD-L1 expression by tumor-infiltrating inflammatory cells multiplied by the density of inflammation (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)).

[0298] 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 status of the tumor is at least about 1%. In another embodiment, the PD-L1 status of the subject is at least about 5%. In a specific embodiment, the PD-L1 status of the tumor is at least about 10%. In one embodiment, the PD-L1 status of the tumor is at least about 25%. In a specific embodiment, the PD-L1 status of the tumor is at least about 50%.

[0299] As used herein, "PD-L1 positive" may be used interchangeably with "at least about 1% PD-L1 expression." In one embodiment, a PD-L1 positive tumor may accordingly have 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 PD-L1 expressing tumor cells when measured by automated IHC. In a specific embodiment, "PD-L1 positive" means that there are at least 100 cells expressing PD-L1 on the surface of the cells. In another embodiment, "PD-L1 positive" means that at least one tumor cell expresses PD-L1 on its surface in a tumor sample containing at least 100 total tumor cells.

[0300] In one embodiment, a tumor derived from NSCLC that is PD-L1 positive and has a high TMB is more likely to respond to a combination therapy comprising (1) an induction phase comprising administering chemotherapy to a subject for a period shorter than the standard duration for that 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), than to a tumor that has only a high TMB, only PD-L1 positive expression, or neither. In one embodiment, the tumor derived from NSCLC has 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% PD-L1 expression. In a specific embodiment, a tumor derived from NSCLC having ≥50% PD-L1 expression and a high TMB status is more likely to respond to a combination therapy using (a) an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an anti-CTLA-4 antibody than a tumor having only high TMB, only ≥50% PD-L1 expression, or neither.

[0301] In certain embodiments, tumors in subjects suitable for the combination therapy disclosed herein do not express PD-L1 (less than 1%, less than 2%, less than 3%, less than 4%, or less than 5% membrane PD-L1). In some embodiments, the method of the present disclosure is not related to PD-L1 expression.

[0302] MSI status

[0303] The TMB status may be used alone or in combination with other factors, e.g., MSI status, as a means of predicting the responsiveness of tumors derived from NSCLC to the combination therapy disclosed herein. In one embodiment, the MSI status is part of the TMB status. In another embodiment, the MSI status is measured separately from the TMB status.

[0304] Microsatellite instability (MSI) is a condition of genetic hypermutability caused by impaired DNA mismatch repair (MMR). The presence of MSI indicates phenotypic evidence that MMR is not functioning normally. In most cases, the genetic basis for instability in MSI tumors is a hereditary wiring alteration in any one of the following five human MMR genes: MSH2, MLH1, MSH6, PMS2, and PMS1. In certain embodiments, tumors derived from NSCLC (e.g., colon tumors) have a high degree of microsatellite instability (MSI-H) and have at least one mutation in the gene MSH2, MLH1, MSH6, PMS2, or PMS1. In other embodiments, subjects receiving tumor treatment within the control group do not have microsatellite instability (MSS or MSI stable) and do not have mutations in the genes MSH2, MLH1, MSH6, PMS2, and PMS1.

[0305] In one embodiment, a subject suitable for the combination therapy disclosed 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 having at least about 30% more unstable MSI biomarkers. In some embodiments, a tumor derived from NSCLC is MSI-H if wiring alterations are detected in at least 2, at least 3, at least 4, or at least 5 MMR genes. In other embodiments, a tumor derived from NSCLC is MSI-H if wiring alterations are detected in at least 30% of 5 or more MMR genes. In some embodiments, wiring alterations in MMR genes are measured by polymerase chain reaction. In other embodiments, a tumor derived from NSCLC is MSI-H if 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.

[0306] Tumors of the present disclosure

[0307] The present disclosure relates to a method for treating a subject suffering from a tumor. The tumor may be any type of tumor or may be a tumor derived from any type of tumor. In some embodiments, the tumor is selected from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastric cancer, esophageal cancer, bladder cancer, lung cancer, and melanoma. In some embodiments, the tumor is derived from lung cancer, renal cell carcinoma, ovarian cancer, colorectal cancer, gastric 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.

[0308] In some embodiments, the tumor is advanced. In certain embodiments, the tumor is locally advanced. In certain embodiments, the tumor is metastatic. In some embodiments, the tumor is refractory. In certain embodiments, the tumor is refractory to one or more prior therapies to treat the tumor. In certain embodiments, the tumor is refractory to one or more standard management therapies to treat the tumor. In certain embodiments, at least one prior therapy includes chemotherapy. In some embodiments, at least one prior therapy includes platinum-based chemotherapy. In other embodiments, the patient has not received prior therapy to treat the tumor, for example, the patient is naive. In some embodiments, the tumor is not refractory. In some embodiments, the tumor is recurrent.

[0309] NSCLC

[0310] NSCLC is a leading cause of cancer death in the United States and worldwide, exceeding the combined total of breast, colorectal, and prostate cancers. In the United States, it is estimated that 228,190 people will be diagnosed with new lung and bronchial cases, and some 159,480 will die from the disease (Siegel et al. (2014) CA Cancer J Clin 64(1):9-29). The majority of patients (approximately 78%) are diagnosed with advanced / recurrent or metastatic disease. Metastasis from lung cancer to the adrenal glands is common, with about 33% of patients having such metastases. Although NSCLC therapy has progressively improved OS, the benefits have plateaued (the median OS for terminal patients is only 1 year). Progression occurred after 1L therapy in almost all of these subjects, and the 5-year survival rate is only 3.6% in a refractory setting. From 2005 to 2009, the overall 5-year relative 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)).

[0311] The present method can treat NSCLC tumors of any stage. In certain embodiments, the tumor originates from NSCLC of any stage. At least seven stages are used for NSCLC: latent stage (hidden stage), stage 0 (carcinoma in situ), stage I, stage II, stage IIIA, stage IIIB, and stage IV. In the latent stage, the cancer cannot be observed by imaging or bronchoscopy. In stage 0, cancer cells are found in the inner layer of the airway.

[0312] In one embodiment, the present method treats stage I non-squamous NSCLC. Stage I NSCLC is divided into stages IA and IB. In stage IA, the tumor is located only within the lung and is 3 centimeters or less. In stage IB, the cancer has not spread to the lymph nodes and meets one or more of the following criteria: 1) the tumor is greater than 3 centimeters and less than or equal to 5 centimeters; 2) the cancer has spread to the main bronchus and is located at least 2 centimeters below where the trachea connects to the bronchus; 3) the cancer has spread to the innermost layer of the membrane covering the lung; or 4) part of the lung has collapsed or pneumonia (inflammation of the lung) has occurred in the area where the trachea connects to the bronchus.

[0313] In another embodiment, the method of the present disclosure treats stage II non-squamous NSCLC. Stage II NSCLC is divided into stages IIA and IIB. In stage IIA, the cancer has either spread to the lymph nodes or has not spread. If the cancer has spread to the lymph nodes, the cancer has spread only to lymph nodes on the same thoracic side as the tumor, and the lymph nodes containing the cancer are located within the lungs or near the bronchi, and meet one or more of the following criteria: 1) the tumor is 5 centimeters or less; 2) the cancer has spread to the main bronchi and is located at least 2 centimeters below where the trachea connects to the bronchi; 3) the cancer has spread to the innermost layer of the membrane covering the lungs; or 4) part of the lung has collapsed or pneumonia (inflammation of the lung) has occurred in the area where the trachea connects to the bronchi. The tumor is also considered IIA if the cancer has not spread to the lymph nodes and meets one or more of the following criteria: 1) the tumor is greater than 5 centimeters and less than or equal to 7 centimeters; 2) The cancer has spread to the main bronchus and is located at least 2 centimeters below where the trachea connects to the bronchus; 3) The cancer has spread to the innermost layer of the membrane covering the lung; or 4) Part of the lung has collapsed or pneumonia (inflammation of the lung) has occurred in the area where the trachea connects to the bronchus. In stage IIB, the cancer may or may not have spread to the lymph nodes. If the cancer has spread to the lymph nodes, it has spread only to lymph nodes on the same thoracic side as the tumor, and the lymph nodes containing the cancer are located within the lung or near the bronchus, and one or more of the following apply: 1) The tumor is greater than 5 centimeters and less than or equal to 7 centimeters; 2) The cancer has spread to the main bronchus and is located at least 2 centimeters below where the trachea connects to the bronchus; 3) The cancer has spread to the innermost layer of the membrane covering the lung; Or 4) Part of the lung has collapsed or pneumonia (inflammation of the lung) has occurred in the area where the trachea connects to the bronchial tubes.A tumor is also considered IIB if the cancer has not spread to the lymph nodes and meets one or more of the following criteria: 1) the tumor is greater than 7 centimeters; 2) the cancer has spread to the main bronchus (at least 2 centimeters below where the trachea connects to the bronchus), the chest wall, the diaphragm, or the nerves controlling the diaphragm; 3) the cancer has spread to the membranes surrounding the heart or the inner layers of the chest wall; 4) the entire lung has collapsed or pneumonia (inflammation of the lung) has occurred; or 5) one or more distinct tumors are present in the same lung lobe.

[0314] In another embodiment, any method of the present disclosure treats stage III non-squamous NSCLC. Stage IIIA is divided into three sections. These three sections are based on 1) the size of the tumor; 2) the location where the tumor is found; and 3) which lymph nodes (if present) have cancer. In the first type of stage IIIA NSCLC, the cancer has spread to lymph nodes on the same thoracic side as the tumor, and the lymph nodes with cancer are located near the sternum or where the trachea enters the lung. Additionally, 1) the tumor may be of any size; 2) part of the lung (where the trachea connects to the trachea) or the entire lung may be in a state of collapse or pneumonia (inflammation of the lung) may occur; and 3) one or more distinct tumors may exist in the same lung lobe; 4) The cancer may spread to any of the following: a) the main bronchus, but not the area where the trachea connects to the bronchus; b) the chest wall; c) the diaphragm and the nerves controlling it; d) the membranes surrounding the lungs or the inner layer of the chest wall; e) the membranes surrounding the heart. In Type 2 of Stage IIIA NSCLC, the cancer has spread to lymph nodes on the same thoracic side as the tumor, and the lymph nodes containing the cancer are located within the lungs or near the bronchi. Additionally, 1) the tumor may be of any size; 2) the entire lung may collapse or develop pneumonia (inflammation of the lung); and 3) one or more distinct tumors may exist in any lung lobe containing the cancer; 4) The cancer may spread to any of the following: a) the main bronchus, but not the area where the trachea connects to the bronchus; b) the chest wall; c) the diaphragm and the nerves controlling it; d) the membranes surrounding the lungs or the inner layer of the chest wall; e) the heart or the membranes surrounding it; f) major blood vessels entering or leaving the heart; g) the trachea; h) the esophagus; i) the nerves controlling the larynx (voice box); j) the sternum (chest bone) or backbone; or k) the keel (where the trachea connects to the bronchus).In Type 3 of Stage IIIA NSCLC, the cancer has not spread to the lymph nodes, the tumor may be of any size, and the cancer has spread to any of the following: a) the heart, b) major blood vessels entering or leaving the heart, c) the trachea, d) the esophagus, e) nerves controlling the larynx (voice box), f) the sternum (chest bone) or backbone, or g) the keel (where the trachea connects to the bronchi). Stage IIIB is divided into two sections based on 1) the size of the tumor, 2) the location where the tumor is found, and 3) which lymph nodes have cancer. In Type 1 of Stage IIIB NSCLC, the cancer has spread to the lymph nodes on the side of the chest opposite the tumor. Additionally: 1) the tumor may be of any size; 2) part of the lung (where the trachea connects to the bronchi) or the entire lung may collapse or develop pneumonia (inflammation of the lung); 3) One or more distinct tumors may be present in any lung lobe with cancer; and 4) the cancer may spread to any of the following: a) the main bronchus, b) the chest wall, c) the diaphragm and the nerves controlling it, d) the membranes surrounding the lungs or the inner layer of the chest wall, e) the heart or the membranes surrounding it, f) major blood vessels entering or leaving the heart, g) the trachea, h) the esophagus, i) the nerves controlling the larynx (voice box), j) the sternum (chest bone) or backbone, or k) the keel (where the trachea connects to the bronchus). In Type 2 of Stage IIIB NSCLC, the cancer has spread to lymph nodes on the same thoracic side as the tumor. The lymph nodes with cancer are located near the sternum or where the bronchus enters the lung.Additionally, 1) the tumor may be of any size; 2) distinct tumors may exist in different lobes of the same lung; and 3) the cancer has spread to any of the following: a) the heart, b) major blood vessels entering or leaving the heart, c) the trachea, d) the esophagus, e) nerves controlling the larynx (voice box), f) the sternum (chest bone) or backbone, or g) the keel (where the trachea connects to the bronchi).

[0315] In some embodiments, the method of the present disclosure treats stage IV non-squamous NSCLC. In stage IV NSCLC, the tumor may be of any size, and the cancer may spread to the lymph nodes. In stage IV NSCLC, one or more of the following apply: 1) one or more tumors are present in both lungs; 2) cancer is found 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.

[0316] In some embodiments, the subject is a non-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 carcinoma cells. In certain embodiments, the subject has non-squamous carcinoma cells.

[0317] Standard management therapy for cancer

[0318] In some embodiments, the methods disclosed herein are used instead of standard management therapy. In certain embodiments, standard management therapy is used in combination with any of the methods disclosed herein. Standard management therapy for different types of cancer is widely 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, publishes the NCCN Guidelines for Clinical Management of Oncology (NCCN GUIDELINES®), which provide detailed and up-to-date information on standard management care for a wide variety of cancers (see [NCCN GUIDELINES® 2014]).

[0319] Colorectal cancer

[0320] In some embodiments, the combination therapy treats 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 reference [Pawlik et al., Dis. Markers 20(4-5): 199-206 (2004)]). In other embodiments, the colorectal cancer has low microsatellite instability (MSI-L).

[0321] 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 visited December 9, 2015)). Most colorectal cancers are adenocarcinomas. Colorectal cancer exists in five stages: stage 0 (carcinoma in situ), stage I, stage II, stage III, and stage IV. Six types of standard treatments are used for colorectal cancer: 1) surgery including local resection, resection of the columella with anastomosis, or resection of the columella 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 disclosure treats colorectal cancer in conjunction with standard management therapy.

[0322] Rectal cancer exists in five stages: stage 0 (carcinoma in situ), stage I, stage II, stage III, and stage IV. Six types of standard treatments are used for rectal cancer: 1) surgery including polypectomy, local resection, resection, radiofrequency ablation, cryosurgery, and pelvic content removal; 2) radiation therapy; 3) chemotherapy; and 4) targeted therapy including monoclonal antibody therapy. In some embodiments, the method of the present disclosure treats rectal cancer in conjunction with standard management therapy.

[0323] lung cancer

[0324] In some embodiments, the combination therapy of the present disclosure treats a tumor 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.

[0325] NSCLC is a leading cause of cancer death in the United States and worldwide, exceeding the combined total of breast, colorectal, and prostate cancers. In the United States, it is estimated that 228,190 people will be diagnosed with new lung and bronchial cases, and some 159,480 will die from the disease (Siegel et al. (2014) CA Cancer J Clin 64(1):9-29). The majority of patients (approximately 78%) are diagnosed with advanced / recurrent or metastatic disease. Metastasis from lung cancer to the adrenal glands is common, with about 33% of patients having such metastases. Although NSCLC therapy has progressively improved OS, the benefits have plateaued (the median OS for terminal patients is only 1 year). Progression occurred after 1L therapy in almost all of these subjects, and the 5-year survival rate is only 3.6% in a refractory setting. From 2005 to 2009, the overall 5-year relative 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)).

[0326] There are seven stages of NSCLC: latent non-small cell lung cancer, stage 0 (carcinoma in situ), stage I, stage II, stage IIIA, stage IIIB, and stage IV. In some embodiments, the combination therapy of the present disclosure treats NSCLC in combination with standard management therapy.

[0327] Additionally, this method can also be combined with the three modalities commonly used to treat NSCLC patients: surgery, radiation therapy (RT), and chemotherapy. 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 chemotherapy is increasingly used both preoperatively and postoperatively. RT can also be used as adjuvant therapy for patients with resectable NSCLC, as a primary local treatment, or as palliative therapy for patients with incurable NSCLC.

[0328] 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 agonists (e.g., cisplatin, carboplatin), taxane agonists (e.g., paclitaxel, albumin-conjugated paclitaxel, and docetaxel), vinorelbine, vinblastine, etoposide, pemetrexed, and gemcitabine, are useful for stage IV NSCLC. Combinations using many of these drugs yield a one-year survival rate of 30% to 40%, which is superior to that of a single agent. Specific targeted therapies have also been developed for the treatment of advanced lung cancer. For example, bevacizumab (Avastin®) is a mAb that blocks vascular endothelial growth factor A (VEGF-A). Erlotinib (TARCEVA®) is a small molecule TKI targeting the epidermal growth factor receptor (EGFR). Crizotinib (XALKORI®) is a small molecule TKI targeting ALK and MET and is used to treat NSCLC in patients with mutated ALK fusion genes. Cetuximab (ERBITUX®) is an mAb targeting EGFR.

[0329] In some embodiments, this method is used to treat subjects with squamous cell NSCLC. In certain embodiments, this method is used in combination with standard management therapy. There are specific unmet needs among patients with squamous cell NSCLC (representing up to 25% of all NSCLCs) because there are few treatment options after first-line (1L) therapy. Single-acting chemotherapy is the standard management after progression using platinum-based dual chemotherapy (Pt-dual therapy), which results in a median OS of approximately 7 months. Docetaxel remains the standard treatment in this line of therapy, and erlotinib may also be used at a lower frequency. Pemetrexed has also been found to provide clinically equivalent efficacy outcomes with significantly fewer side effects compared to docetaxel in second-line (2L) treatment of patients with advanced NSCLC (Hanna et al., 2004 J Clin Oncol 22:1589-97). There are currently no therapies approved for use in lung cancer after third-line (3L) setting. Pemetrexed and bevacizumab are not approved for squamous NSCLC, and molecularly targeted therapies have limited coverage. Unmet needs in advanced lung cancer have recently worsened as Oncothyreon and Merck KgaA’s STIMUVAX® failed to improve OS in Phase 3 trials, ArQule and Daiichi Sankyo’s c-Met kinase inhibitor tivantinib failed to meet survival endpoints, Eli Lilly’s Alimta® in combination with Roche’s Avastin® failed to improve OS in late studies, and Amgen and Takeda Pharmaceutical failed to meet clinical endpoints using the small molecule VEGF-R antagonist motesanib in late studies.

[0330] Certain aspects of the present disclosure relate to a method of administering chemotherapy for a period shorter than the standard duration for said chemotherapy. In some embodiments, the standard duration for said chemotherapy is based on the standard management regimen for said cancer type. In certain embodiments, the tumor is derived from NSCLC, and the chemotherapy is administered for a period shorter than the standard duration for administering the standard management regimen for the treatment of NSCLC, e.g., stage IV NSCLC.

[0331] Standard management regimens for different types of cancer are widely 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, publishes the NCCN Guidelines for Clinical Management of Oncology (NCCN GUIDELINES®), which provide detailed and up-to-date information on standard management treatments for a wide variety of cancers (see [NCCN GUIDELINES® (2018)] (the full text of which is incorporated herein by reference), available at www.nccn.org / professionals / physician_gls / default.aspx (last accessed October 22, 2018).

[0332] Among patients with squamous cell NSCLC (accounting for up to 25% of all NSCLCs), there are specific unmet needs due to the scarcity of treatment options following first-line (1L) therapy. Mono-agonist chemotherapy is the standard management after progression using platinum-based dual chemotherapy (Pt-dual therapy), which results in a median OS of approximately 7 months. Docetaxel remains the standard treatment in this line of therapy, while erlotinib may also be used at a lower frequency. Pemetrexed has also been shown to provide clinically equivalent efficacy outcomes compared to docetaxel in second-line (2L) treatment of patients with advanced NSCLC, accompanied by significantly fewer adverse effects (Hanna et al. (2004) J Clin Oncol 22:1589-97).

[0333] The NCCN guidelines for the treatment of NSCLC with chemotherapy include, but are not limited to, treatments selected from the following: (i) cisplatin 75 mg / m² 2 Day 1 plus pemetrexed 500 mg / m² 2 Day 1, for non-squamous cells every 21 days during 4 cycles; (ii) carboplatin AUC 6 Day 1, paclitaxel 200 mg / m² 2 Day 1, every 21 days for 4 cycles; and (iii) carboplatin AUC 5 Day 1, pemetrexed 500 mg / m² 2 Day 1, for non-squamous cells every 21 days for 4 cycles. Refer to [NCCN Guidelines Version 6.2018 Non-Small Cell Lung Cancer]. Other standard management chemotherapy regimens include (iv) cisplatin 50 mg / m² 2 Day 1 and Day 8 and vinorelbine 25 mg / m² 2 Day 1, Day 8, Day 15, Day 22, every 28 days during 4 cycles; (v) cisplatin 100 mg / m² 2 Day 1 and vinorelbine 30 mg / m² 2Day 1, Day 8, Day 15, Day 22, every 28 days during 4 cycles; (vi) cisplatin 75-80 mg / m² 2 Day 1 and vinorelbine 25-30 mg / m² 2 Days 1 and 8, every 21 days during 4 cycles; (vii) cisplatin 100 mg / m² 2 Day 1 and etoposide 100 mg / m² 2 Days 1–3, every 28 days for 4 cycles; (viii) cisplatin 75 mg / m² 2 Day 1 and gemcitabine 1250 mg / m² 2 Days 1 and 8, every 21 days during 4 cycles; (ix) cisplatin 75 mg / m² 2 Day 1 and docetaxel 75 mg / m² 2 Day 1, every 21 days for 4 cycles; and (x) carboplatin AUC 5 Day 1, gemcitabine 1000 mg / m² 2 Day 1 and Day 8, including every 21 days during 4 cycles. Accordingly, in some embodiments, the method of the present disclosure includes an induction step comprising administering chemotherapy, e.g., standard management chemotherapy, for a period shorter than the standard duration for that chemotherapy. In certain embodiments, the duration is less than 4 cycles. In some embodiments, the duration is less than 3 cycles. In some embodiments, the duration is less than 2 cycles. In certain embodiments, the duration is 2 cycles.

[0334] In some embodiments, the subject has received one or more prior therapies for the tumor. In certain embodiments, at least one prior therapy comprises a standard management regimen for the treatment of 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 some embodiments, at least one prior therapy is selected from a regimen comprising the administration of an anticancer agent selected from the group consisting of platinum agonists (e.g., cisplatin, carboplatin), taxane agonists (e.g., paclitaxel, albumin-conjugated paclitaxel, docetaxel), vinorelbine, vinblastine, etoposide, pemetrexed, gemcitabine, bevacizumab (Avastin®), erlotinib (Tarceva®), crizotinib (Zalkori®), cetuximab (Erbitux®), and any combination thereof. In certain embodiments, at least one prior therapy comprises platinum-based dual chemotherapy.

[0335] In some embodiments, the subject experienced disease progression after at least one prior therapy. In certain embodiments, the subject 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 received at least two prior therapies. In one embodiment, the subject experienced disease progression after at least two prior therapies. In certain embodiments, at least two prior therapies include a first prior therapy and a second prior therapy, wherein the subject 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 wherein the second prior therapy includes surgery, radiation therapy, chemotherapy, immunotherapy, or any combination thereof. In some embodiments, the first prior therapy includes platinum-based dual chemotherapy, and the second prior therapy includes single-acting chemotherapy. In certain embodiments, the single-acting chemotherapy includes docetaxel.

[0336] In some aspects of the present disclosure, the method disclosed herein further comprises administering additional anticancer therapy. The additional anticancer therapy may include any therapy known in the art for the treatment of NSCLC or tumors derived therefrom and / or any standard management therapy as disclosed herein. In some embodiments, the additional anticancer therapy includes surgery, radiation therapy, additional chemotherapy, additional immunotherapy, or any combination thereof. In some embodiments, the additional anticancer therapy includes additional chemotherapy including any chemotherapy disclosed herein. In some embodiments, the additional anticancer therapy includes additional immunotherapy. In some embodiments, the additional anticancer therapy includes administration of an antibody or its antigen-binding portion 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.

[0337] In certain embodiments, additional anticancer therapy is administered in conjunction with, and subsequently with, the administration of an anti-PD-1 antibody (or anti-PD-L1 antibody) and an anti-CTLA-4 antibody, or in conjunction with and subsequently with them. In some embodiments, additional anticancer therapy is administered in conjunction with the administration of an anti-PD-1 antibody (or anti-PD-L1 antibody) and an anti-CTLA-4 antibody. In some embodiments, additional anticancer therapy is administered after the administration of an anti-PD-1 antibody (or anti-PD-L1 antibody) and an anti-CTLA-4 antibody. In some embodiments, additional anticancer therapy is administered in conjunction with and subsequently with the administration of an anti-PD-1 antibody (or anti-PD-L1 antibody) and an anti-CTLA-4 antibody. In other embodiments, additional anticancer therapy is administered between the anti-PD-1 antibody (or anti-PD-L1 antibody) and the anti-CTLA-4 antibody. In certain embodiments, additional anticancer therapy, an anti-PD-1 antibody (or anti-PD-L1 antibody) and / or an anti-CTLA-4 antibody are combined into a single agent. In other embodiments, additional anticancer therapy, an anti-PD-1 antibody (or anti-PD-L1 antibody) and / or an anti-CTLA-4 antibody are present as individual agents.

[0338] Melanoma

[0339] In some embodiments, combination therapy treats tumors derived from melanoma. Melanoma is the deadliest form of skin cancer, 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 visited December 9, 2015)). Melanoma exists in seven stages: stage 0 (melanoma in situ), stage I, stage II, surgically removable stage III, surgically inmovable stage III, stage IV, and recurrent melanoma. Five standard types of treatment are used: 1) surgery; 2) chemotherapy; 3) radiation therapy and 4) biological therapies including interferon, interleukin-2 (IL-2), tumor necrosis factor (TNF) therapy and ipilimumab, and 5) signaling inhibitor therapy (e.g., vemurafenib, dabrafenib and trametinib), oncolytic virus therapy, monoclonal antibody therapy (including pembrolizumab and nivolumab), and targeted therapies including angiogenesis inhibitors. In some embodiments, the combination therapy of the present disclosure treats melanoma in combination with standard management therapy.

[0340] ovarian cancer

[0341] In certain embodiments, combination therapy treats tumors derived from ovarian cancer, fallopian tube cancer, and / or primary peritoneal cancer ("ovarian cancer"). In certain embodiments, the cancer is ovarian epithelial carcinoma. In other embodiments, the cancer is an ovarian germ cell tumor. In yet another embodiment, the cancer is an ovarian low-malignancy potential tumor. In some embodiments, ovarian cancer originates in tissues covering the ovary, peritoneum, or fallopian tube (see http: / / www.cancer.gov / types / ovarian / patient / ovarian-epithelial-treatment-pdq (last accessed December 9, 2015)).

[0342] There are four stages of ovarian cancer: stages I, II, III, and IV exist, which encompass early, advanced, and recurrent or persistent ovarian cancer. There are four types of standard treatments used for patients with ovarian cancer, salpingo-oophorectomy, and primary peritoneal cancer: 1) surgery including hysterectomy, unilateral salpingo-oophorectomy, bilateral salpingo-oophorectomy, retinal resection, and lymph node biopsy; 2) radiation therapy; 3) chemotherapy; and 4) targeted therapy including monoclonal antibody therapy and poly(ADP-ribose) polymerase inhibitors. Biological therapies are also being tested for ovarian cancer. In some embodiments, the combination therapy of the present disclosure treats ovarian cancer in conjunction with standard management therapy.

[0343] There are four stages of ovarian germ cell tumors: stages I, II, III, and IV. Four types of standard treatment are used: 1) surgery including unilateral salpingo-oophorectomy, total hysterectomy, bilateral salpingo-oophorectomy, and tumor volume reduction; 2) observation; 3) chemotherapy; and 4) radiation therapy. Novel treatment options under consideration include high-dose chemotherapy accompanied by bone marrow transplantation. In some embodiments, the combination therapy of the present disclosure treats ovarian germ cell tumors in conjunction with standard management therapy.

[0344] There are three stages of ovarian low-malignant potential tumors: 1) early (stages I and II), 2) late (stages III and IB), and 3) recurrent. Two types of standard treatments are used: 1) surgery including unilateral salpingo-oophorectomy, bilateral salpingo-oophorectomy, total hysterectomy, partial oophorectomy, and retinaculomy, and 2) chemotherapy. In some embodiments, the combination therapy of the present disclosure treats ovarian low-malignant potential tumors in combination with standard management therapy.

[0345] Head and neck cancer

[0346] In some embodiments, combination therapy treats cancer that is head and neck cancer. Head and neck cancer includes cancer of the oral cavity, pharynx, larynx, sinuses, nasal cavity, and salivary glands. Head and neck cancer typically originates in squamous cells present in the inner layer of the moist mucosal surface within the head and neck (e.g., within the oral cavity, nasal cavity, and throat). These squamous cell carcinomas are often referred to as squamous cell carcinomas of the head and neck. Head and neck cancer can also originate in the salivary glands, but salivary gland cancer is relatively rare (see http: / / www.cancer.gov / types / head-and-neck / head-neck-fact-sheet (last accessed December 9, 2015)). A treatment plan for an individual patient depends on a number of factors, including the exact location of the tumor, the stage of the cancer, and the person's age and overall health. Treatment for head and neck cancer may include surgery, radiation therapy, chemotherapy, targeted therapy, or a combination of these therapies. In some embodiments, the combination therapy of the present disclosure treats head and neck cancer in combination with standard management therapy.

[0347] Pharmaceutical composition and dosage

[0348] The therapeutic agent of the present disclosure may consist of a pharmaceutical composition containing, for example, an antibody and / or cytokine and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption retardants, etc. that are physiologically compatible. Preferably, the carrier for the composition containing the antibody is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), whereas the carrier for the composition containing the antibody and / or cytokine is suitable for parenteral, e.g., 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 full text of which is incorporated herein by reference). Enhanz® uses a co-preparation of an antibody and a recombinant human hyaluronidase enzyme (rHuPH20), which overcomes traditional limitations on the volume of bioagents and drugs that can be delivered subcutaneously due to the extracellular matrix (see U.S. Patent No. 7,767,429). The pharmaceutical composition of the present disclosure may comprise one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or ajuvants, such as preservatives, wetting agents, emulsifiers, and dispersants. Accordingly, in some embodiments, the pharmaceutical composition of the present disclosure may further comprise a recombinant human hyaluronidase enzyme, e.g., rHuPH20.

[0349] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody is administered as a single composition at a fixed dose together with the anti-CTLA-4 antibody. In some embodiments, the anti-PD-1 antibody is administered as a single composition at a fixed dose together with the anti-CTLA-4 antibody. In some embodiments, the anti-PD-L1 antibody is administered as a single composition at a fixed dose together with the anti-CTLA-4 antibody. 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 It is 100:1, approximately 90:1, approximately 80:1, approximately 70:1, approximately 60:1, approximately 50:1, approximately 40:1, approximately 30:1, approximately 20:1, approximately 15:1, approximately 10:1, approximately 9:1, approximately 8:1, approximately 7:1, approximately 6:1, approximately 5:1, approximately 4:1, approximately 3:1, or approximately 2:1 mg.

[0350] Although higher nivolumab monotherapy with up to 10 mg / kg administered every two weeks was achieved without reaching the maximum allowable dose (MTD), significant toxicity reported in other trials of checkpoint inhibitor plus anti-angiogenesis therapy (e.g., see literature [Johnson et al., 2013; Rini et al., 2011]) supports selecting a nivolumab dose of less than 10 mg / kg.

[0351] Treatment is continued as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in certain embodiments, the dose of the anti-PD-1 antibody, anti-PD-L1 antibody and / or anti-CTLA-4 antibody administered is significantly lower than the approved dose, i.e., an agent below the therapeutic dose is administered. The anti-PD-1 antibody, anti-PD-L1 antibody and / or anti-CTLA-4 antibody may be administered at the dose presented as producing the highest efficacy as monotherapy in clinical trials, for example, about 3 mg / kg of nivolumab may be administered once every 3 weeks (Topalian et al., 2012a; Topalian et al., 2012), or at a significantly lower dose, i.e., below the therapeutic dose.

[0352] The dosage and frequency depend on the half-life of the antibody in the subject. Generally, human antibodies have the longest half-lives, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. The dosage and frequency may vary depending on whether the treatment is prophylactic or therapeutic. For prophylactic purposes, relatively low doses are typically administered over a long period at relatively infrequent intervals. Some patients continue treatment for the rest of their lives. For therapeutic purposes, relatively high doses at relatively short intervals are sometimes required until disease progression is reduced or terminated, preferably until the patient shows partial or complete improvement in disease symptoms. After that, prophylactic therapy may be administered to the patient.

[0353] The actual dosage level of the active ingredient in the pharmaceutical composition of the present disclosure may vary to obtain an amount of the active ingredient effective in achieving the desired therapeutic response for a specific patient, composition, and mode of administration without being excessively toxic to the patient. The selected dosage level will depend on various pharmacokinetic factors, including the activity of the specific composition of the present disclosure used, the route of administration, the time of administration, the elimination rate of the specific compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the specific composition used, the age, sex, weight, condition, general health, and medical history of the patient to be treated, and other factors widely known in the medical field. The composition of the present disclosure may be administered via one or more routes of administration using one or more of the various methods widely known in the relevant art. As will be recognized by a person skilled in the art, the route and / or mode of administration will vary depending on the desired result.

[0354] Kit

[0355] A kit comprising (a) a chemotherapy agent, (b) an anti-PD-1 antibody or an anti-PD-L1 antibody, and (c) an anti-CTLA-4 antibody for therapeutic use is also within the scope of this disclosure. The kit typically includes a label and instructions for use indicating the intended use of the contents of the kit. The term "label" includes any document or record supplied on or with the kit, or otherwise enclosed with the kit. In some embodiments, this disclosure comprises (a) a dose of carboplatin sufficient to administer an AUC 6 dose and 200 mg / m² 2(a) a dose of paclitaxel; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody in a dose ranging from 200 mg to 800 mg or a dose ranging from 200 mg to 1800 mg; (c) an anti-CTLA-4 antibody in a dose ranging from 0.1 to 10 mg / kg body weight; (d) instructions for using (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 method disclosed herein. In some embodiments, the present disclosure provides (a) a dose of carboplatin sufficient to administer a dose of AUC 5 and 500 mg / m² 2 (a) a dosage of pemetrexed; (b) an anti-PD-1 antibody or an anti-PD-L1 antibody in a dosage ranging from 200 mg to 800 mg or 200 mg to 1800 mg; (c) an anti-CTLA-4 antibody in a dosage ranging from 0.1 to 10 mg / kg body weight; (d) instructions for using (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 method disclosed herein. In some embodiments, the present disclosure provides (a) a dosage of carboplatin sufficient to administer a dose of AUC 6 and 500 mg / m² 2The present disclosure provides a kit for treating a subject suffering from a tumor, e.g., a tumor derived from NSCLC, comprising (a) a dose of pemetrexed; (b) a dose of an anti-PD-1 antibody or an anti-PD-L1 antibody in a dose ranging from 200 mg to 800 mg; (c) a dose of an anti-CTLA-4 antibody in a dose ranging from 0.1 to 10 mg / kg body weight; and (d) instructions for using (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 method disclosed herein. In some embodiments, the present disclosure provides (a) 75 mg / m² 2 cisplatin at a dosage of 500 mg / m² 2 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 pemetrexed; (b) a dose of an anti-PD-1 antibody or an anti-PD-L1 antibody in a range of 200 mg to 800 mg; (c) a dose of an anti-CTLA-4 antibody in a range of 0.1 to 10 mg / kg body weight; and (d) instructions for using (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 method disclosed herein.

[0356] In a specific preferred embodiment for treating a human patient, the kit comprises an anti-human PD-1 antibody disclosed herein, e.g., nivolumab or pembrolizumab. In a specific preferred embodiment for treating a human patient, the kit comprises an anti-human PD-L1 antibody disclosed herein, e.g., atezolizumab, durvalumab or avelumab. In a specific preferred embodiment for treating a human patient, the kit comprises an anti-human CTLA-4 antibody disclosed herein, e.g., ipilimumab, tremelimumab, MK-1308 or AGEN-1884.

[0357] 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.

[0358] In some embodiments, the kit further comprises the comprehensive genome profiling assay disclosed herein. In some embodiments, the kit comprises the Foundation One® CDX™ genome profiling assay. In some embodiments, 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 Mb in at least about 10 mutations / sequencing genomes, according to the method disclosed herein. In other embodiments, 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 Mb in at least about 10 mutations / sequencing genomes, according to the method disclosed herein.

[0359] In certain embodiments, the kit further includes a assay for detecting and / or measuring tumor cell surface PD-L1 expression. In certain embodiments, the kit further includes the Dako PD-L1 IHC 28-8 pharmDx test for IHC staining of tumor cell surface PD-L1 protein.

[0360] All references cited above, as well as all references cited herein, are incorporated herein by reference in their entirety.

[0361] The following examples are provided as examples without limitation.

[0362] Examples

[0363] Example 1: Study of nivolumab combined with ipilimumab as a first-line therapy for the treatment of stage IV non-small cell lung cancer (NSCLC)

[0364] A clinical trial is underway to investigate the safety and efficacy of a combination of nivolumab and ipilimumab as a first-line treatment for stage IV NSCLC. Nivolumab is administered in combination with ipilimumab, which is administered intravenously at 1 mg / kg over 30 minutes every 6 weeks, or intravenously at 3 mg / kg over 30 minutes every 2 weeks, until disease progression, unacceptable toxicity, or other prior-specified reasons occur. Treatment with nivolumab and ipilimumab will be provided for up to 2 years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0365] purpose

[0366] The primary objectives of this study are (i) to determine the objective response rate (ORR) in all treated PD-L1 positive (membrane staining in ≥ 1% of tumor cells) subjects treated with nivolumab in combination with ipilimumab as first-line therapy by a blinded, independent central-institutional review according to RECIST 1.1; and (ii) to determine the ORR in all treated PD-L1 negative (≤ 1%) subjects treated with nivolumab in combination with ipilimumab as first-line therapy by a blinded, independent central-institutional review according to RECIST 1.1.

[0367] Secondary objectives include (i) evaluating ORR by a blinded, independent central agency review according to RECIST 1.1 in all treated subjects treated with nivolumab in combination with ipilimumab as first-line therapy; (ii) evaluating progression-free survival (PFS) based on a blinded, independent central agency review assessment according to RECIST 1.1; (iii) evaluating overall survival; (iv) evaluating ORR, PFS, and OS by PD-L1 expression levels; and (v) evaluating tumor mutation burden (TMB) as a potential predictive biomarker of the efficacy (e.g., ORR, PFS, and OS) of nivolumab in combination with ipilimumab using DNA derived from tumor specimens.

[0368] Exploratory objectives include (i) evaluating the safety and tolerability, pharmacokinetics and immunogenicity of nivolumab combined with ipilimumab as a first-line therapy; and (ii) evaluating tumor inflammatory gene expression signatures as potential predictive biomarkers of the efficacy (e.g., ORR, PFS and OS) of nivolumab combined with ipilimumab using RNA derived from tumor specimens.

[0369] Research Design

[0370] 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 combination therapy. Subjects will receive nivolumab 3 mg / kg once every 2 weeks and ipilimumab 1 mg / kg once every 6 weeks. On the day of the infusion, nivolumab will be administered first. The second infusion will always be ipilimumab (if scheduled to be provided) and will begin at least 30 minutes after the completion of the nivolumab infusion. Subjects will receive combination therapy until disease progression or unacceptable toxicity, or for up to 2 years.

[0371] The use of topical, ocular, intra-articular, intranasal, and inhaled corticosteroids (with minimal systemic absorption) is permitted for subjects. Adrenal replacement steroid doses of prednisone > 10 mg per day are permitted. Short (less than 3 weeks) courses of corticosteroids are permitted for prophylaxis (e.g., contrast dye allergy) or for the treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions induced by contact allergens). Regular concomitant use of bisphosphonates and RANK-L inhibitors for the prevention or reduction of skeletal-related events in patients with bone metastases is permitted if initiated prior to the first dose of the study regimen. Prior palliative radiotherapy must have been completed at least 2 weeks prior to treatment.

[0372] Inclusion / Exclusion Criteria

[0373] Eligible subjects must have histologically confirmed stage IV NSCLC (according to the 7th International Association for Lung Cancer Research classification) with squamous or non-squamous histology and disease measurable by CT or MRI according to RECIST 1.1 criteria. Subjects must not have received prior systemic therapy for stage IV disease. Prior definitive chemoradiotherapy for locally advanced disease is permitted provided that the last administration of chemotherapy or radiotherapy (whichever was last provided) occurred at least 6 months prior to enrollment. Locally advanced disease with recurrence after co-chemoradiotherapy (stage IIIB, specifically referring to patients with no curative treatment options) is eligible for enrollment. Prior adjuvant or neoadjuvant chemotherapy for early lung cancer is permitted if completed at least 6 months prior to the initiation of study treatment. Subjects must be EGFR / ALK wild-type and have an EXOG PS of 0 or 1. Tumor tissue samples must be available for PD-L1 immunohistochemistry (IHC) testing.

[0374] Formalin-fixed, paraffin-embedded (FFPE) tissue blocks or unstained tumor tissue sections must be submitted for biomarker evaluation prior to treatment, along with a report on the associated pathology. Tumor tissue samples may be fresh or stored if obtained within six months prior to enrollment, and no systemic therapy (e.g., adjuvant or neoadjuvant chemotherapy) may have been administered after the sample was obtained. Tissue samples must be collected by core needle biopsy, excision, or incisional biopsy. Fine needle biopsy or drainage of pleural effusion using cytospin is not considered appropriate for biomarker review. Biopsies of bone lesions lacking soft tissue components or decalcified bone tumor samples are also not accepted.

[0375] Subjects with known EGFR mutations susceptible to available targeted inhibitor therapy (including, but not limited to, deletions in exon 19 and exon 21 [L858R] substitution mutations) are excluded. All subjects with non-squamous histology must have been tested for EGFR mutation status. EGFR testing must be performed locally. The use of FDA-approved or local health agency-approved tests is strongly recommended. Tests other than PCR or next-generation sequencing will be required to be repeated using PCR or next-generation sequencing-based methods. Subjects with non-squamous histology having unknown or indeterminate EGFR status are excluded.

[0376] Subjects with known ALK potentials susceptible to available targeted inhibitor therapy are excluded. ALK testing must be performed at a local laboratory, and the use of FDA-approved testing is strongly recommended. Subjects with unknown or indeterminate ALK status may be enrolled.

[0377] Subjects with untreated CNS metastases are excluded. Subjects are eligible if the CNS metastases are adequately treated and the subject returns to neurological baseline for at least 2 weeks prior to the first dose (excluding residual signs or symptoms associated with CNS treatment). Additionally, the subject must discontinue corticosteroids or receive a stable or decreasing dose of ≤ 10 mg of prednisone (or equivalent) daily for at least 2 weeks prior to the first dose.

[0378] Subjects with carcinomatous meningitis or active known or suspected autoimmune diseases are excluded. Subjects with type I diabetes mellitus, hypothyroidism requiring only hormone replacement, skin disorders not requiring systemic treatment (e.g., vitiligo, psoriasis, or alopecia), or conditions not expected to recur in the absence of external triggers are permitted for enrollment.

[0379] Subjects with a condition requiring systemic treatment with corticosteroids (1 mg prednisone equivalent per day) or other immunosuppressive drugs within 14 days of the first treatment are excluded. Inhaled or topical steroids and adrenal replacement steroids (1 mg prednisone equivalent per day) are permitted in the absence of active autoimmune disease.

[0380] Subjects with a history of failing a screening for any anti-PD-L1 or anti-PD-L1 antibody clinical trial due to a PD-L1-negative status are also excluded.

[0381] Research evaluation

[0382] Subjects will be evaluated for PD-L1 expression in tumor cells and will be categorized into four groups (PD-L1 positive, PD-L1 negative, PD-L1 ≥ 50%, and PD-L1 indeterminate). PD-L1 status will be determined by the Daco PD-L1 IHC 28-8 pharmDx test for IHC staining of PD-L1 protein in submitted tumor samples. PD-L1 positive is characterized by ≥ 1% tumor cell membrane staining in at least 100 evaluable tumor cells. PD-L1 negative 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. PD-L1 indivisibility is defined as subjects lacking quantifiable PD-L1 expression at baseline, which may be due to insufficient tumor biopsy specimens for IHC staining and analysis. For subjects with PD-L1 indivisible tumors, key efficacy and safety parameters will be summarized in a descriptive manner.

[0383] Tumor evaluation during the study by CT or MRI will begin 6 weeks after the first day of administration (+ / - 7 days) and will be performed every 6 weeks (+ / - 7 days) until 48 weeks. After 48 weeks, tumor evaluation will be performed every 12 weeks (+ / - 7 days) until disease progression is recorded or treatment is discontinued (whichever happens later).

[0384] In subjects receiving treatment containing nivolumab and ipilimumab, if the subject tolerates nivolumab plus ipilimumab with investigator-assessed clinical benefit, treatment after initial RECIST 1.1-defined progression is permitted. Subjects receiving investigational treatment after investigator-assessed progression must also continue tumor evaluation until such treatment is discontinued.

[0385] OS will be tracked continuously while the subject is receiving the study drug and every three months through face-to-face or telephone contact after the subject has stopped receiving the study drug.

[0386] terminal

[0387] The primary endpoint is (i) the objective response rate (ORR) in all treated PD-L1 positive (≥1%) subjects in stage IV NSCLC subjects treated with nivolumab in combination with ipilimumab as first-line therapy, based on a blinded, independent central agency review according to RECIST 1.1; and (ii) the ORR in all treated PD-L1 negative (<1%) subjects in stage IV NSCLC subjects treated with nivolumab in combination with ipilimumab as first-line therapy, based on a blinded, independent central agency review according to RECIST 1.1.

[0388] ORR is based on a blind, independent central agency review evaluation according to RECIST 1.1 criteria. ORR is defined as the number of subjects with a confirmed CR or PR (BOR) divided by the number of treated subjects among PD-L1 positive or PD-L1 negative subjects, or the total number of treated subjects. BOR is defined as the designation of the best response recorded between baseline and the objectively recorded date of progression according to RECIST 1.1, or the date of initiation of palliative local therapy or subsequent chemotherapy (whichever occurs first).

[0389] PFS determined by a blind, independent central review is defined as the time from the first dosing date to the treated subject to the first recorded date of tumor progression or death from any cause as determined by a blind, independent central review (in accordance with RECIST 1.1). Subjects who have not progressed or died will be censored on their last evaluable tumor evaluation date. Subjects who have not undergone any study tumor evaluation will be censored on the first dosing date. Subjects who have initiated any palliative local therapy or subsequent chemotherapy without previously reported progression will be censored on their last evaluable tumor evaluation date prior to the initiation of palliative local therapy or subsequent chemotherapy (whichever procedure occurs first).

[0390] Secondary endpoints are (i) ORR based on a blinded, independent central agency review according to RECIST 1.1 in all treated subjects treated with nivolumab in combination with ipilimumab as first-line therapy; (ii) PFS based on a blinded, independent central agency review assessment according to RECIST 1.1; (iii) OS; (iv) ORR, PFS, and OS based on PD-L1 expression levels; and (iv) total somatic mutations in tumor cells and their association with ORR, PFS, and OS. OS is defined as the time between the first day of administration and the day of death from any cause.

[0391] Example 2: Study of nivolumab plus ipilimumab combined with chemotherapy as a first-line therapy for the treatment of stage IV non-small cell lung cancer (NSCLC)

[0392] A clinical trial is underway to investigate the safety and tolerability of phase-in first-line treatment for stage IV NSCLC using a combination of nivolumab, ipilimumab, and chemotherapy. As induction therapy, nivolumab is administered intravenously, ipilimumab is administered intravenously along with two cycles of histology-based platinum dual chemotherapy, followed by nivolumab and ipilimumab until disease progression or unacceptable toxicity occurs. Treatment with nivolumab and ipilimumab will be provided for up to two years from the start of the study treatment in the absence of disease progression or unacceptable toxicity.

[0393] Two cycles of histology-based platinum dual chemotherapy consist of carboplatin AUC6 plus paclitaxel 200 mg / m² for squamous histology. 2 , and for non-squamous histology (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m² 2 or (ii) cisplatin 75 mg / m² 2 + Pemetrexed 500 mg / m² 2 Includes

[0394] purpose

[0395] The primary objectives of this study are (i) to determine the incidence of DLT (dose-limiting toxicity) during the DLT evaluation period (within 9 weeks after the first dose); and (ii) to determine the safety and tolerability of nivolumab and ipilimumab in combination with chemotherapy.

[0396] The secondary objective is to evaluate ORR, PFS, and OS by researcher review using RECIST 1.1.

[0397] The exploratory objectives include (i) assessing overall health status and lung cancer symptoms, as measured by the Lung Cancer Symptom Scale (LCSS) Mean Symptom Burden Index (ASBI), using the EQ-5D descriptive system and visual analog scales in subjects receiving nivolumab plus chemotherapy combined with ipilimumab; (ii) evaluating tumor inflammatory gene expression signatures as potential predictive biomarkers of the efficacy (e.g., ORR, PFS, and OS) of nivolumab plus ipilimumab combined with chemotherapy using RNA derived from tumor specimens; and (iii) exploring TMB as a potential predictive biomarker of the efficacy (e.g., ORR, PFS, and OS) of nivolumab plus ipilimumab combined with chemotherapy using DNA derived from tumor specimens.

[0398] Research Design

[0399] A safety induction phase will be performed to evaluate safe dose levels. Approximately 28 subjects (to achieve at least 22 DLT-evaluable subjects) will receive two cycles of induction chemotherapy and nivolumab plus ipilimumab (Fig. 1). The starting dose of nivolumab is 360 mg every 3 weeks, and ipilimumab is 1 mg / kg every 6 weeks. Nivolumab will be administered in combination with ipilimumab plus two cycles of histology-based platinum dual chemotherapy. For squamous histology, carboplatin AUC6 plus paclitaxel 200 mg / m² 2 will be administered. For non-squamous histology, (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m² 2 or (ii) cisplatin 75 mg / m² 2 Plus Pemetrexed 500 mg / m² 2 will be administered.

[0400] After two cycles of induction therapy, nivolumab is administered intravenously over 30 minutes in combination with ipilimumab, until disease progression, unacceptable toxicity, or other reasons specified in the protocol. Treatment with nivolumab and ipilimumab will be provided for up to two years from the start of study treatment in the absence of disease progression or unacceptable toxicity.

[0401] "Safe" is defined as 25% or fewer of the evaluated subjects exhibiting DLT (i.e., 5 or fewer subjects among 22 DLT-evaluable subjects having such an event).

[0402] A safety assessment will be performed on the first 10 subjects after a minimum of 9 weeks of follow-up. If 20% or fewer of the first 10 subjects exhibit DLT (i.e., 2 or fewer subjects with such events), the regimen is determined to be safe, and enrollment for a subsequent study using this combination may begin while the safety introduction phase is in progress. If more than 20% of the first 10 subjects exhibit DLT, the entire safety cohort will be evaluated before using this dose regimen in a subsequent study. If more than 25% of the total DLT-evaluable subjects exhibit DLT, the protocol may be modified to evaluate different dose levels based on observed toxicity.

[0403] After the above two cycles of induction therapy, nivolumab and ipilimumab will be continued until disease progression or unacceptable toxicity, or withdrawal of consent, or for up to two years from the start of study therapy in the absence of disease progression or unacceptable toxicity.

[0404] During the safety introduction phase, subjects are permitted to use topical, ocular, intra-articular, intranasal, and inhaled corticosteroids (with minimal systemic absorption). Adrenal replacement steroid doses of >10 mg prednisone per day are permitted. Short (less than 3 weeks) courses of corticosteroids are permitted for prophylaxis (e.g., contrast dye allergy) or for the treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions induced by contact allergens). Regular concomitant use of bisphosphonates and RANK-L inhibitors for the prevention or reduction of skeletal-related events in patients with bone metastases is permitted if initiated prior to the first dose of the study regimen. Prior palliative radiotherapy must have been completed at least 2 weeks prior to treatment.

[0405] Inclusion / Exclusion Criteria and Research Evaluation

[0406] The inclusion and exclusion criteria and study evaluation are the same as in Example 1, except that the tumor tissue sample may be fresh or stored if obtained within 3 months prior to registration.

[0407] terminal

[0408] The primary endpoints are (i) determining the incidence of DLT within 9 weeks after the first dose; and (ii) determining the safety and tolerability of nivolumab and ipilimumab in combination with chemotherapy. The secondary endpoints are evaluating ORR, PFS, and OS by investigator assessment using RECIST 1.1. OS is defined as the time between the first dose date and the date of death from any cause.

[0409] PFS determined by a blind, independent central review is defined as the time from the first dosing date to the treated subject to the first recorded date of tumor progression or death from any cause as determined by a blind, independent central review (in accordance with RECIST 1.1). Subjects who have not progressed or died will be censored on their last evaluable tumor evaluation date. Subjects who have not undergone any study tumor evaluation will be censored on the first dosing date. Subjects who have initiated any palliative local therapy or subsequent chemotherapy without previously reported progression will be censored on their last evaluable tumor evaluation date prior to the initiation of palliative local therapy or subsequent chemotherapy (whichever procedure occurs first).

[0410] Secondary endpoints are (i) ORR based on a blinded, independent central review according to RECIST 1.1 in all treated subjects treated with nivolumab in combination with ipilimumab as first-line therapy; (ii) PFS based on a blinded, independent central review assessment according to RECIST 1.1; (iii) OS; (iv) ORR, PFS, and OS based on PD-L1 expression levels; and (iv) total somatic mutations in tumor cells and their association with ORR, PFS, and OS.

[0411] Preliminary results

[0412] Thirty-six participants were treated with nivolumab plus ipilimumab combined with chemotherapy for a minimum follow-up of 11 weeks (maximum follow-up of 9 months, median follow-up of 4.7 months). There was one case of dose-limiting toxicity (DLT) defined by elevated liver enzymes. Twenty-five percent of participants experienced drug-related SAEs, and 8 percent of participants experienced treatment-related discontinuation of nivolumab, ipilimumab, and chemotherapy. No safety concerns were observed during ongoing monitoring.

[0413] Dose-limiting toxicity was defined as any of the following occurring during the first 9 weeks: (i) any Grade 2 drug-related uveitis or ophthalmic pain unresponsive to topical therapy, not improving to Grade 1 severity within the retreatment period, or requiring systemic treatment; (ii) any Grade 2 drug-related pneumonia or interstitial lung disease not resolving within 14 days with dose delay and systemic steroids (radiological changes may take longer to resolve); (iii) any Grade 3 non-cutaneous drug-related adverse events, excluding laboratory abnormalities, that are not remission (defined as return to Grade 1, radiological changes may take longer to resolve) or control by appropriate management (appropriate management is defined as the treatment outlined in the AE management algorithm in the investigator's brochure) within 14 days; (iv) any Grade 4 drug-related adverse events, including Grade 4 leukopenia or neutropenia lasting less than 14 days and laboratory abnormalities, excluding asymptomatic amylase / lipase evaluation; and (v) any of the following drug-related liver function laboratory abnormalities: > AST or ALT > 5-10x ULN, AST or ALT > 10x ULN, total bilirubin > 5x ULN, combined AST or ALT > 3x ULN and total bilirubin > 2x ULN for 2 weeks, and grade 3 thrombocytopenia associated with bleeding.

[0414] Example 3: Phase 3 randomized study of nivolumab plus ipilimumab combined with chemotherapy as first-line therapy in stage IV NSCLC

[0415] Initial evaluation of tolerability data from the safety introduction study of Example 2 indicated that two cycles of platinum-based chemotherapy with nivolumab 360 mg every 3 weeks and ipilimumab 1 mg / kg every 6 weeks were safe and had no DLT after at least 9 weeks of follow-up. This randomized phase 3 study evaluates safety in a larger cohort of patients and compares the efficacy and other outcomes of nivolumab plus ipilimumab combined with chemotherapy with those of chemotherapy alone.

[0416] purpose

[0417] The primary objective of this study is to compare the efficacy of nivolumab plus ipilimumab combined with chemotherapy against chemotherapy in participants with histologically confirmed stage IV NSCLC. Secondary objectives include (i) evaluating efficacy outcomes in participants with histologically confirmed stage IV NSCLC treated with nivolumab plus ipilimumab combined with chemotherapy by different PD-L1 expression levels compared to chemotherapy; and (ii) evaluating the tumor mutation burden as a potential predictive biomarker for the efficacy (e.g., ORR, PFS, and OS) of nivolumab plus ipilimumab combined with chemotherapy.

[0418] The exploratory objectives are to (i) evaluate candidate predictive biomarkers, including but not limited to intratumoral as well as peripheral biomarkers, as potential predictive biomarkers for the efficacy of nivolumab plus ipilimumab combined with chemotherapy; (ii) evaluate the safety and tolerability of nivolumab plus ipilimumab combined with chemotherapy; (iii) characterize the immunogenic potential of nivolumab and ipilimumab; (iv) characterize the pharmacokinetics of nivolumab and ipilimumab; (v) evaluate overall health status using the EQ-5D descriptive system and visual analogy scales in participants receiving nivolumab combined with chemotherapy plus ipilimumab and participants receiving platinum dual chemotherapy; (vi) evaluate the disease-related symptom burden in participants with histologically confirmed stage IV NSCLC treated with nivolumab plus ipilimumab combined with chemotherapy compared to chemotherapy alone; and (vii) evaluate the participants' healthcare resource utilization (HCRU). and (viii) to evaluate the efficacy of nivolumab plus ipilimumab combined with chemotherapy compared to chemotherapy in participants with histologically confirmed stage IV NSCLC after the next course of treatment.

[0419] Research Design

[0420] This study consists of two divisions: a treatment division and a control division (Fig. 2). In the treatment division, subjects receive induction therapy of intravenous nivolumab and intravenous ipilimumab, followed by two cycles of histology-based platinum dual chemotherapy, and then nivolumab and ipilimumab until disease progression or unacceptable toxicity occurs. Treatment with nivolumab and ipilimumab will be provided for up to 24 months in the absence of disease progression or unacceptable toxicity. Subjects in the treatment division receive 360 ​​mg of nivolumab every 3 weeks and 1 mg / kg of ipilimumab every 6 weeks. The two cycles of histology-based platinum dual chemotherapy are histology-based. Subjects with squamous histology receive carboplatin AUC 6 plus paclitaxel 200 mg / m² 2 Subjects with non-squamous histology receive (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m² 2 or (ii) cisplatin 75 mg / m² 2 + Pemetrexed 500 mg / m² 2 The patient is administered cisplatin. Prior to randomization, the investigator must determine whether a participant with non-squamous histology will receive cisplatin if they are eligible. Chemotherapy is administered on the 1st day of each 3-week cycle, for example, on the 1st day of the first 3-week cycle and again on the 1st day of the second 3-week cycle (Table 8). After two cycles of induction therapy, nivolumab 360 mg every 3 weeks and ipilimumab 1 mg / kg every 6 weeks are continued until progression, unacceptable toxicity, or other predetermined reason.

[0421] Table 8: Administered treatment.

[0422]

[0423] Both nivolumab and ipilimumab should be administered as a 30-minute infusion. Nivolumab is administered first. Ipilimumab is administered at least 30 minutes after the completion of the nivolumab infusion. Platinum-dual therapy begins at least 30 minutes after the completion of the nivolumab or ipilimumab infusion (if scheduled to be administered ipilimumab). In the investigator's judgment, if a prior infusion reaction occurs in the participant, nivolumab or ipilimumab may be administered over a longer infusion time (60 minutes).

[0424] Participants are permitted to use topical, ocular, intra-articular, intranasal, and inhaled corticosteroids (with minimal systemic absorption). Adrenal replacement steroid doses of >10 mg prednisone per day are permitted. Short (less than 3 weeks) courses of corticosteroids are permitted for prophylaxis (e.g., contrast dye allergy) or for the treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions induced by contact allergens). Regular concomitant use of bisphosphonates and RANK-L inhibitors for the prevention or reduction of skeletal-related events in patients with bone metastases is permitted if initiated prior to the first dose of the study regimen. Prior palliative radiotherapy must have been completed at least 2 weeks prior to treatment.

[0425] Paclitaxel and Carboplatin

[0426] The participant received paclitaxel 200 mg / m² as a 180-minute IV infusion. 2It will be received on the 1st day of the 3-week cycle with a dose of carboplatin at an AUC of 6 as a 30-minute IV infusion, or at doses according to local prescribing information. Infusion times may follow local clinical trial standards. Paclitaxel dosage calculations must be based on body surface area calculations. Doses may be maintained if the participant's body weight is within 10% of baseline weight or previous dose weight. Oral corticosteroids should be provided according to local standards at a dose equivalent to dexamethasone 20 mg 12 and 6 hours prior to paclitaxel administration. Oral or intravenous diphenhydramine (or its equivalent) 50 mg and an H2-blocker (according to local standard administration) should be administered 30 to 60 minutes prior to the paclitaxel infusion. The doses of paclitaxel and / or carboplatin may be stopped, delayed, reduced, or discontinued depending on how well the participant tolerates treatment.

[0427] Carboplatin should be administered on the first day of each cycle following paclitaxel, and the carboplatin dose will be calculated using the following Calvert formula: the carboplatin dose (mg) is equal to the target AUC x ([CrCl (ml / min) + 25]). The calculation of creatinine clearance (CrCl) is based on the Cockcroft-Gault formula and should include the most recent serum creatinine and most recent body weight. Note that if the calculation of CrCl by the Cockcroft-Gault formula yields a result > 125 mL / min, CrCl should be calculated by an alternative formula in accordance with clinical trial standards or capped at 125 mL / min. The dose of carboplatin may be capped according to local standards.

[0428] Pemetrexed and cisplatin

[0429] The calculation of pemetrexed dosage should be based on body surface area calculations. The dosage may be maintained if the participant's body weight is within 10% of the body weight used to calculate the previous dose. Oral corticosteroids should be administered according to local standards at a dose equivalent to dexamethasone 4 mg BID on the day before, the day of, and the day after pemetrexed administration. Oral folic acid 350 to 1000 mcg daily should be administered starting one week prior to the first dose of pemetrexed, and at least five doses of folic acid should be administered seven days prior to the first dose. Oral folic acid should be continued daily throughout the treatment with pemetrexed and for 21 days after the last dose of pemetrexed. Intramuscular (IM) injection of vitamin B12 1000 mcg should be administered approximately one week prior to the first dose of pemetrexed and thereafter repeated every three cycles during pemetrexed treatment. A follow-up injection of Vitamin B12 may be administered on the same day as pemetrexed (participants with non-squamous histology may anticipate pemetrexed and begin folic acid and Vitamin B12 prior to randomization). Pre-metabolic antiemetics will be administered according to local standards. Recommended antiemetic treatments are dexamethasone (administration according to local standards; may be replaced with another corticosteroid of equivalent dose) and 5-HT3 receptor antagonists (type according to investigator's judgment and local standard administration). Additional use of pre-metabolic antiemetics may be at the investigator's judgment.

[0430] Participants received 75 mg / m² on the first day of a 3-week treatment cycle according to local standards for up to 2 cycles in the treatment division or 4 cycles in the control division. 2 500 mg / m² as a 10-minute IV infusion on Day 1 with infusion dose of cisplatin 2 You will receive a dose of Pemetrexed.

[0431] Cisplatin will be administered to the participant at least 30 minutes after the completion of the pemetrexed infusion. Pre-treatment hydration for cisplatin may follow local standard management, or the use of 1 to 2 liters of fluid administered intravenously 8 to 12 hours prior to the cisplatin infusion (according to local standards) is recommended. Adequate hydration and urine output must be maintained for at least 24 hours after cisplatin administration. Administration and monitoring should be performed according to local standards. The use of mannitol after the cisplatin infusion should also follow local standard management.

[0432] Participants discontinuing cisplatin monotherapy may be switched to pemetrexed / carboplatin for the remaining platinum-based dual therapy cycles (up to 2 cycles in the treatment section or a total of 4 cycles in the control section), at the investigator's discretion. Administration of pemetrexed / carboplatin to these participants must follow the following pemetrexed / carboplatin guidelines, with or without the pemetrexed continuation maintenance section.

[0433] Pemetrexed and Carboplatin

[0434] Participants received 500 mg / m² as a 10-minute IV infusion on the first day of a 3-week treatment cycle for up to 4 cycles. 2 Following the dose of pemetrexed, on the first day, a dose of carboplatin with an AUC of 5 or 6 will be received as a 30-minute IV infusion. The dose of carboplatin will be calculated as described above.

[0435] Control group

[0436] In the control group, participants receive 4 cycles of histology-based platinum chemotherapy. Subjects exhibiting squamous histology receive carboplatin AUC 6 plus paclitaxel 200 mg / m² 2 Subjects with non-squamous histology receive (i) carboplatin AUC 5 or 6 plus pemetrexed 500 mg / m² 2or (ii) cisplatin 75 mg / m² 2 + Pemetrexed 500 mg / m² 2 Receives cisplatin. Prior to randomization, the investigator must determine whether or not a participant with non-squamous histology will receive cisplatin if eligible. Chemotherapy is administered on Day 1 of each 3-week cycle. Participants with non-squamous histology who have stable disease or a response after induction chemotherapy may receive any pemetrexed maintenance therapy: 500 mg / m² on Day 1 of each 3-week cycle until disease progression, unacceptable toxicity, or other predetermined reason. 2 It is permitted to receive Pemetrexed.

[0437] PD-L1 expression

[0438] Participants will be evaluated by PD-L1 expression and categorized into three groups (PD-L1 positive, PD-L1 negative, and PD-L1 indivisible). PD-L1 status will be determined by the Daco PD-L1 IHC 28-8 pharmDx test for IHC staining of PD-L1 protein in submitted tumor samples. PD-L1 positive is characterized by ≥ 1% tumor cell membrane staining in at least 100 evaluable tumor cells. PD-L1 negative is characterized by < 1% tumor cell membrane staining in at least 100 evaluable tumor cells. Participants with PD-L1 indivisible have tumor biopsy specimens lacking quantifiable PD-L1 expression. Patients with tested but indivisible PD-L1 will be stratified as PD-L1 negative participants. The PD-L1 indivisible group will be capped at 10% of the total randomized population.

[0439] Dose reduction for chemotherapy

[0440] A reduction in the dose of chemotherapy may be required and will be performed in accordance with Table 9. The reduction in the dose of chemotherapy is permanent; once the dose of any chemotherapy agent is reduced, it cannot be increased again in subsequent cycles, except as mentioned when starting pemetrexed maintenance therapy. The reduction in dose for each agent in platinum-based dual chemotherapy regimens is not associated and can be adjusted independently as summarized below.

[0441] Table 9: Dose adjustment of chemotherapy agents

[0442]

[0443] Research group

[0444] Approximately 700 patients will be randomized in a 1:1 ratio between the treatment and control groups. Participants must have histologically confirmed stage IV non-small cell lung cancer (NSCLC), squamous or non-squamous histology, as defined by the 7th International Association for Lung Cancer Research classification. Subjects must not have received prior systemic therapy for stage IV disease. Subjects must be EGFR / ALK wild-type and have an ECOG performance status of ≤ 1. Participants must have tumor tissue samples for biomarker analysis, and participants must have undergone a PD-L1 IHC trial with results available for randomization, or the PD-L1 trial will be performed by a central laboratory. Prior definitive chemoradiotherapy for locally advanced disease is permitted provided that the last administration of chemotherapy or radiotherapy (whichever was last administered) occurred at least 6 months prior to enrollment. Locally advanced disease with recurrence after chemoradiotherapy (stage IIIB disease, specifically referring to patients with no cure options) is eligible for enrollment. Neoadjuvant or neoadjuvant chemotherapy for early-stage lung cancer is permitted if completed at least 6 months prior to the initiation of study treatment. Neoadjuvant palliative radiotherapy for non-CNS lesions must have been completed at least 2 weeks prior to treatment. Subjects with symptomatic tumor lesions at baseline who may require palliative radiotherapy within 4 weeks of the first treatment are strongly encouraged to receive palliative radiotherapy before treatment.

[0445] Subjects will be excluded for any of the following reasons. Participants with known EGFR mutations susceptible to available targeted inhibitor therapy (including, but not limited to, deletions in exon 19 and exon 21 [L858R] substitution mutations) are excluded. All participants with non-squamous histology must have been tested for EGFR mutation status. EGFR testing must be performed locally. EGFR testing is not provided by third-party laboratories. The use of FDA-approved or local health agency-approved testing is strongly recommended. Participants with non-squamous histology having an unknown or indeterminate EGFR status are excluded. Participants with known ALK translocations susceptible to available targeted inhibitor therapy are excluded. If tested, the use of FDA-approved testing is strongly recommended. Participants with an unknown or indeterminate ALK status may be enrolled. Participants with untreated CNS metastases are excluded. Participants are eligible if CNS metastases are adequately treated and the participant returns to neurological baseline for at least 2 weeks prior to the first treatment (excluding residual signs or symptoms associated with CNS treatment). Additionally, participants must discontinue corticosteroids or receive a stable or decreasing dose of ≤ 10 mg prednisone (or equivalent) daily for at least 2 weeks prior to the first treatment. Participants with carcinomatous meningitis will be excluded. Participants with a prior malignancy (excluding non-melanoma skin cancer and carcinoma in situ, e.g., bladder cancer, gastric cancer, colon cancer, cervical cancer / dysplasia, melanoma, or breast cancer) are excluded unless complete remission was achieved at least 2 years prior to the first treatment and no additional therapy is required or expected to be required during the study period. Participants with an active, known, or suspected autoimmune disease.Participants with type I diabetes mellitus, hypothyroidism requiring only hormone replacement, skin disorders not requiring systemic treatment (e.g., vitiligo, psoriasis, or alopecia), or conditions not expected to recur in the absence of external triggers are permitted to enroll. Subjects with conditions requiring systemic treatment with corticosteroids (1 mg prednisone equivalent > 10 mg daily) or other immunosuppressive drugs within 14 days of the first treatment are excluded. Inhaled or topical steroids and adrenal replacement steroids (1 mg prednisone equivalent > 10 mg daily) are permitted in the absence of active autoimmune disease.

[0446] terminal

[0447] The primary endpoint is determining overall survival (OS). Additional endpoints are (i) progression-free survival (PFS) and objective response rate (ORR); (ii) OS, PFS, and ORR for participants with different PD-L1 levels; (iii) the total number of somatic mutations in tumor cells and their association with ORR, PFS, and OS; (iv) gene expression signatures (e.g., tumor-inflammatory gene expression signatures, driver mutations, immune cell infiltration, tumor inflammation, immune cell infiltration, etc.), driver mutations (e.g., STK11, KRAS), as well as peripheral markers and soluble factors in the blood (e.g., cytokines, solHLA, soluble inflammatory / immunosuppressive factors) and other factors in the blood (e.g., MDSC, miRNA) and their association with clinical outcomes (ORR, PFS, and OS); (v) blood TMB analysis using plasma circulating free DNA; and (vi) incidence rates of adverse events (AEs), sub-epithelial events (SAEs), and selective AEs. (vii) relationship between anti-nivolumab and anti-ipilimumab antibodies and other outcome measures; (viii) pharmacokinetic (PK) measures of nivolumab and ipilimumab; (ix) EQ-5D-3L descriptive system and visual analog scale (EQ-5D VAS); (x) Lung cancer symptom score (LCSS) mean symptom burden index (ASBI); (xi) incidence of HCRU; and (xii) PFS after the next course of treatment (PFS2).

[0448] biomarkers

[0449] Tumor tissue specimen

[0450] Storage (or fresh) FFPE tumor tissues must be collected within 3 months prior to enrollment. Where feasible, random fresh biopsies may also be collected at baseline or at the time of clinical progression. Samples collected at progression can provide important information regarding the underlying mechanisms of the biology of acquired resistance and are therefore highly valuable in these exploratory studies.

[0451] Tissue samples must be sent to a central laboratory for determination of PD-L1 status using an analytically validated IHC assay. PD-L1 stained tissue samples will be evaluated by a pathologist at the central laboratory confirmed by the sponsor, and will be scored as PD-L1 expression if membrane staining is observed in ≥ 1% of tumor cells out of at least 100 evaluable tumor cells.

[0452] To explore the potential association between tumor mutation burden and clinical outcomes, tumor tissue will be evaluated by targeted and / or whole-exome sequencing. Gene expression signatures, such as those associated with non-limiting inflammatory processes and / or immune-related signaling, will also be evaluated by targeted and / or whole-transcriptome RNA sequencing for their potential association with clinical outcomes.

[0453] Tissues may also be analyzed by IHC or similar methodologies to determine the abundance of immunomodulatory proteins, such as PD-L1, PD-L2, PD-1, and other markers associated with TILs (e.g., CD4, CD8, FOXP3), to a non-limiting extent. These data will be evaluated for potential associations with clinical endpoints.

[0454] Single nucleotide polymorphism (SNP)

[0455] After whole blood is collected from all participants, it will be processed to generate genomic DNA for SNP analysis. These analyses will focus on SNPs within genes associated with PD-1 and other immunomodulatory signaling pathways to determine whether natural variations within these genes are associated with the response to nivolumab and / or adverse events during treatment. Genomic DNA derived from whole blood can also be used as a wiring control for whole-exome sequencing studies within tumors.

[0456] Serum Soluble Factor

[0457] To understand the prevalence of circulating proteins and their potential impact on the clinical activity of nivolumab, protein concentrations of a panel of cytokines, chemokines, and other related immunomodulatory, serum-soluble factors (e.g., soluble PD-L1) will be investigated at baseline and during treatment.

[0458] Serum microRNA (miRNA)

[0459] MicroRNAs (miRNAs) are widely expressed small RNAs that regulate the abundance of mRNA transcripts and their translation into proteins. Overall miRNA expression profiling is becoming increasingly common in cancer research, and miRNA signatures correlated with disease stage or clinical outcomes are currently available for various cancer types. MiRNA expression profiling can also be useful for identifying molecular markers for predicting drug responses and for prospective stratification. Interestingly, miRNAs may be stable in serum and may indicate overexpression in tumors or reflect immune system activity. Serum samples taken from participants randomized to their respective treatment divisions at baseline and during treatment will be analyzed for miRNA content using microarrays or similar methodologies. The generated miRNA profiles will be evaluated for changes in miRNA abundance occurring after treatment and their association with response and survival data. Ultimately, the goal will be to determine whether unique immune-related and / or NSCLC-related miRNA signatures exist and whether they are potentially useful in identifying patients who may (or may not) respond to nivolumab treatment.

[0460] Bone marrow-derived suppressor cells (MDSC)

[0461] Myeloid-derived suppressor cells are a population of immune cells capable of inhibiting T cell activation and proliferation. Low pre-treatment levels of myeloid-derived suppressor cells (MDSCs) in peripheral blood may be associated with superior overall survival (OS) in melanoma patients treated with the immunotherapy agent ipilimumab. MDSCs will be measured at baseline / pre-treatment and during treatment to assess their association with outcomes.

[0462] Peripheral blood mononuclear cells (PBMC)

[0463] Peripheral blood mononuclear cells in whole blood taken from participants at baseline and during treatment will be analyzed by flow cytometry or other methods (e.g., ELIspot) to evaluate immune cell activity.

[0464] Blood TMB using circulating plasma / cell-free DNA (cfDNA)

[0465] Circulating / cell-free DNA (cfDNA) is a small fragment of DNA that leaks from tumors and non-malignant cells and can be found circulating in the peripheral bloodstream. cfDNA derived from malignant cells can be isolated from blood-derived plasma and analyzed for genetic features, including but not limited to somatic mutations, using targeted sequencing-based methods (e.g., next-generation sequencing). To complement the planned genomic analysis outlined for tumor evaluation, plasma will be collected at baseline and during treatment to isolate cfDNA. Mutation data derived from these samples will be compared with those directly identified in the tumor. Changes in mutation burden (overall or individual genes) at baseline and during treatment will be evaluated for their association with treatment outcomes. The correlation between the mutation burden in the blood and the tumor will be explored.

[0466] On October 21, 2019, it was announced that the clinical trial segment of this study met its primary endpoint of superior overall survival (OS) in a pre-specified interim analysis. The comparator in this study was randomized maintenance therapy following chemotherapy alone for four cycles or less. In this study, the safety profile of nivolumab plus low-dose ipilimumab and two cycles of chemotherapy reflected the known safety profiles of immunotherapy and chemotherapy components in first-line NSCLC.

Claims

Claim 1 A pharmaceutical composition comprising nivolumab for use in a method for treating a tumor in a subject suffering from non-small cell lung cancer (NSCLC), wherein the method comprises: (1) administering to the subject on the first day of a first 3-week cycle (a) a pharmaceutical composition administered at a dose of 360 mg nivolumab, (b) ipilimumab administered at a dose of 1 mg / kg, and (c) a platinum dual chemotherapy comprising (i) carboplatin and paclitaxel, (ii) pemetrexed and cisplatin, or (iii) pemetrexed and carboplatin; (2) administering to the subject on the first day of a second 3-week cycle (a) a pharmaceutical composition administered at a dose of 360 mg nivolumab and (b) said platinum dual chemotherapy; and (3) subsequently, (2) administer (i) the above pharmaceutical composition and (ii) ipilimumab to a subject, wherein nivolumab is administered at a dose of 360 mg every 3 weeks and ipilimumab is administered at a dose of 1 mg / kg every 6 weeks, pharmaceutical composition. Claim 2 In claim 1, the platinum dual chemotherapy consists of carboplatin AUC 6 and paclitaxel 200 mg / m² 2 A pharmaceutical composition comprising on the first day of each three-week cycle. Claim 3 In paragraph 1, the platinum dual chemotherapy consists of carboplatin AUC 5 or AUC 6 and pemetrexed 500 mg / m² 2 A pharmaceutical composition comprising Claim 4 In paragraph 1, the platinum double chemotherapy is cisplatin 75 mg / m² 2 and pemetrexed 500 mg / m² 2 A pharmaceutical composition comprising Claim 5 A pharmaceutical composition according to claim 1, wherein the subject exhibits progression-free survival for at least 1 month or at least 2 months after administration. Claim 6 A pharmaceutical composition according to claim 1, wherein the subject exhibits total survival of at least one month after administration. Claim 7 A pharmaceutical composition according to claim 1, wherein the subject exhibits an objective reaction rate of at least 15%. Claim 8 A pharmaceutical composition according to claim 1, wherein the tumor has a TMB state in which the tumor has a high tumor mutation burden (TMB). Claim 9 A pharmaceutical composition according to claim 8, wherein the TMB state is determined by sequencing nucleic acids in a biological sample obtained from a subject and confirming genomic alterations in the sequenced nucleic acids. Claim 10 A pharmaceutical composition according to claim 9, wherein the genomic alteration comprises (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 changes (CNAs), gene rearrangements, and any combination thereof; or (v) any combination of (i)-(iv). Claim 11 A pharmaceutical composition according to claim 8, wherein the high TMB has a score of at least 210. Claim 12 A pharmaceutical composition according to claim 8, wherein the TMB state of the subject is compared with a reference TMB value, wherein the TMB state of the subject is within the highest quartile of the reference TMB value or wherein the TMB state of the subject is within the upper quartile of the reference TMB value. Claim 13 A pharmaceutical composition according to claim 9, 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. Claim 14 A pharmaceutical composition according to claim 8, wherein the TMB state is determined by (i) genome sequencing, (ii) exome sequencing, (iii) genome profiling, or (iv) any combination of (i)-(iii). Claim 15 제14항에 있어서, 게놈 프로파일이 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 (C17orf39), 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), C11orf30 (EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1T1, TERT (프로모터만), 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, CCND2, DAXX, FGF23, GSK3B, KMT2A (MLL), NF1, POLD1, SETD2, TOP1, ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C (MLL3), NF2POLE, SF3B1, TOP2A, ARID2, CCNE1, 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, CDC73, 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, 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, BCL6, CDKN2C, ESR1, FUBP1,A pharmaceutical composition comprising one or more genes selected from the group consisting of 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 any combination thereof. Claim 16 A pharmaceutical composition according to claim 8, wherein (i) the tumor comprises a non-small cell carcinoma, (ii) the tumor is relapsed or refractory after at least one prior regimen for treating the tumor, or (iii) both (i) and (ii). Claim 17 A pharmaceutical composition according to claim 14, wherein the genome profile includes mutation testing. Claim 18 A pharmaceutical composition according to claim 8, wherein the tumor has a TMB of at least 10 mutations per megabase of the sequenced genome. Claim 19 A pharmaceutical composition according to claim 1, wherein the tumor is derived from stage IV NSCLC. Claim 20 In claim 19, a pharmaceutical composition in which the NSCLC is flat NSCLC. Claim 21 In claim 19, a pharmaceutical composition in which the NSCLC is non-flat NSCLC. Claim 22 A pharmaceutical composition according to claim 1, wherein the tumor is locally advanced, progressive, or metastatic. Claim 23 A pharmaceutical composition according to claim 1, wherein the tumor is refractory or recurrent. Claim 24 A pharmaceutical composition according to claim 1, wherein the tumor is refractory after at least one prior therapy for treating the tumor, wherein at least one prior therapy comprises a platinum agonist, cisplatin, carboplatin, a taxane agonist, paclitaxel, albumin-conjugated paclitaxel, docetaxel, vinorelbine, vinblastine, etoposide, pemetrexed, gemcitabine, bevacizumab (AVASTIN®), erlotinib (TARCEVA®), crizotinib (XALKORI®), cetuximab (ERBITUX®), or any combination thereof. Claim 25 A pharmaceutical composition according to claim 1, wherein at least 1% of the tumor cells exhibit membrane PD-L1 expression. Claim 26 A pharmaceutical composition according to claim 1, wherein at least 5% of the tumor cells exhibit membrane PD-L1 expression. 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