Combination therapy using anti-Fucosyl-GM1 antibodies

Abstract

The present disclosure provides combination therapies for treating a subject, e.g., a subject suffering from lung cancer, such as small cell lung cancer, comprising administering to the subject various combinations of an anti-Fucosyl-GM1 antibody and an immunomodulatory agent, e.g., a PD-1 / PD-L1 antagonist, e.g., an anti-PD-1 or anti-PD-L1 antagonist antibody, carboplatin and etoposide.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 63 / 135,479, filed January 8, 2021, the entirety of which is hereby incorporated by reference as part of this specification.

[0002] Sequence Listing The sequence listing submitted electronically with this application is also incorporated by reference in its entirety as part of this specification (Filename: 20220104_SEQL_13935WOPCT_GB; Creation Date: January 4, 2022; File Size: 20KB).

[0003] FIELD OF THEINVENTION The present invention relates to an improved method for the treatment of small cell lung cancer comprising the administration of a combination of an antibody against Fucosyl-GM1, an antibody against PD-1 or PD-L1, and a chemotherapy regimen. [Background technology]

[0004] 2. Background of the Invention Fucosyl GM1 is a sphingolipid monosialoganglioside consisting of a ceramide lipid moiety that anchors the molecule to the cell membrane and a carbohydrate moiety that is exposed on the cell surface. Carbohydrate antigens are the most abundantly expressed antigens on the surface of cancer cells (Feizi T. (1985) Nature 314:53-7). In some cancer types, such as small cell lung cancer (SCLC), the initial response to chemotherapy is remarkable but relapses occur quickly. Novel immunotherapeutic interventions can be successful in overcoming drug-resistant recurrences (Johnson DH. (1995) Lung Cancer 12Suppl 3:S71-5). Some carbohydrate antigens, such as gangliosides GD3 and GD2, have been shown to function as effective targets for passive immunotherapy with mAbs (Irie RF and Morton DL (1986) PNAS 83:8694-8698; Houghton AN et al. (1985) PNAS 82:1242-1246). Ganglioside antigens have also been shown to be effective targets for active immunotherapy with vaccines in clinical trials (Krug LM et al. (2004) Clinical Cancer Research 10:6094-6100; Dickler MN et al. (1999) Clinical Cancer Research 5:2773-2779; Livingston PO et al. (1994) J. Clin. Oncol. 12:1036-44). Indeed, sera from SCLC patients who developed antibody titers against Fucosyl-GM1 after vaccination with KLH-conjugated antigen showed specific binding to tumor cells and tumor-specific complement-dependent cytotoxicity (CDC). Anti-Fucosyl-GM1 antibody titer-associated toxicity was mild and transient, and three patients with limited-stage SCLC were relapse-free at 18, 24, and 30 months (Krug et al., supra; Dickler et al., supra).

[0005] Fucosyl-GM1 expression has been demonstrated in a high percentage of SCLC cases, and unlike other ganglioside antigens, fucosyl-GM1 is expressed little or not at all in normal tissues (Nilsson et al. (1984) Glycoconjugate J. 1:43-9; Krug et al., supra; Brezicka et al. (1989) Cancer Res. 49:1300-5; Zhangyi et al. (1997) Int. J. Cancer 73:42-49; Brezicka et al. (2000) Lung Cancer 28:29-36; Fredman et al. (1986) Biochim. Biophys. Acta 875:316-23; Brezicka et al. (1991) APMIS 99:797-802; Nilsson et al. (1986) Cancer Res. 46:1403-7). Fucosyl-GM1 has been demonstrated to be present in culture medium from SCLC cell lines, in tumor extracts and in serum from nude mouse xenografts, as well as in serum from SCLC patients with extensive disease (Vangsted et al. (1991) Cancer Res. 51:2879-84; Vangsted et al. (1994) Cancer Detect. Prev. 18:221-9). Fucosyl-GM1 expression has also been observed in a significant proportion of non-small cell lung cancer (NSCLC) samples. WO07 / 067992. These reports provide convincing evidence that Fucosyl-GM1 is a highly specific tumor antigen that can be targeted by immunotherapy. Summary of the Invention [Problem to be solved by the invention]

[0006] Anti-Fucosyl-GM1 mAb BMS-986012, an antibody that recognizes Fucosyl-GM1 on cancer cells and induces its destruction, has entered clinical trials for the treatment of patients with relapsed / refractory small cell lung cancer (NCT02247349). See Molckovsky & Siu (2008) J. Hematol. Oncol. 1:20. BMS-986012 is a non-fucosylated antibody and therefore exhibits enhanced ADCC compared to antibodies with typical mammalian glycosylation. Although effective as a single agent, there is a need for more effective lung cancer therapies. [Means for solving the problem]

[0007] SUMMARY OF THE PRESENT APPLICATION The present invention provides a combination therapy for the treatment of lung cancer, particularly SCLC, comprising an initial round, e.g., four rounds, of induction therapy, optionally followed by one or more rounds of maintenance therapy, where the induction therapy comprises treatment with carboplatin, etoposide, an anti-Fucosyl GM1 antibody and an anti-PD-1 / PD-L1 antibody, and the maintenance therapy comprises treatment with an anti-Fucosyl GM1 antibody and an anti-PD-1 / PD-L1 antibody.

[0008] In some embodiments, the anti-Fucosyl-GM1 mAb competes with BMS-986012, contains the same CDRs as BMS-986012, contains the same heavy and light chain variable domains as BMS-986012, contains the same heavy and light chains as BMS-986012, is BMS-986012, or is an antibody drug conjugate of BMS-986012. In one embodiment, the immunomodulatory agent is nivolumab (OPDIVO (登録商標) In yet another embodiment, the immunomodulatory agent is an anti-PD-1 mAb that competes with nivolumab, contains the same CDRs as nivolumab, contains the same heavy and light chain variable domains as nivolumab, or is nivolumab. (登録商標)In yet another embodiment, the immunomodulatory agent is an anti-PD-1 mAb that competes with pembrolizumab, contains the same CDRs as pembrolizumab, contains the same heavy and light chain variable domains as pembrolizumab, or is pembrolizumab. (登録商標) In a further embodiment, the immunomodulatory agent is an anti-PD-1 mAb that competes with cemiplimab-rwlc, contains the same CDRs as cemiplimab-rwlc, contains the same heavy and light chain variable domains as cemiplimab-rwlc, or is cemiplimab-rwlc. (登録商標) In a further embodiment, the immunomodulatory agent is an anti-PD-L1 mAb that competes with atezolibumab, contains the same CDRs as atezolibumab, contains the same heavy and light chain variable domains as atezolibumab, or is atezolibumab. (登録商標) In additional embodiments, the immunomodulatory agent is an anti-PD-L1 mAb that competes with durvalumab, contains the same CDRs as durvalumab, contains the same heavy and light chain variable domains as durvalumab, or is durvalumab. (登録商標) ), contains the same CDRs as avelumab, contains the same heavy and light chain variable domains as avelumab, or is avelumab.

[0009] In some embodiments, each round of induction therapy is for 21 days (Q3W). In some embodiments, the induction therapy includes treatment with carboplatin administered intravenously (iv) at an area under the curve (AUC) of 5 mg / ml / min on day 1 of each cycle of induction therapy. In an alternative embodiment, carboplatin is replaced by cisplatin at 80 mg / ml. 2 In some embodiments, the induction therapy may be administered at 100 mg / m on days 1, 2, and 3 of each cycle of induction therapy. 2In some embodiments, the induction therapy includes treatment with an anti-fucosyl-GM1 mAb, such as BMS-986012, administered at 420 mg iv on day 1 of each cycle of induction therapy. In some embodiments, the induction therapy includes treatment with an anti-PD-1 mAb, such as nivolumab, administered at 360 mg iv on day 1 of each cycle of induction therapy. In some embodiments, all four therapeutic agents are administered during induction therapy as described in this paragraph.

[0010] In some embodiments, each round of maintenance therapy is for 28 days (Q4W). In some embodiments, maintenance therapy comprises treatment with an anti-fucosyl-GM1 mAb, e.g., BMS-986012, administered at 560 mg iv on day 1 of each cycle of maintenance therapy. In some embodiments, maintenance therapy comprises treatment with an anti-PD-1 mAb, e.g., nivolumab, administered at 480 mg iv on day 1 of each cycle of maintenance therapy. In some embodiments, both therapeutic agents are administered during maintenance therapy as described in this paragraph.

[0011] In another aspect, the invention provides a method for treating a subject with small cell lung cancer (SCLC), e.g., a subject with extensive-stage small cell lung cancer (ES-SCLC), comprising administering to the subject a therapeutically effective combination of Fucosyl-GM1 and an agent, such as a monoclonal antibody or antigen-binding portion thereof, that specifically binds to an immune-modulatory target, such as PD-1 or PD-L1. In some embodiments, the anti-Fucosyl-GM1 mAb is administered 400 mg or 1000 mg Q3W or Q4W, the anti-PD-1 mAb is administered 360 mg or 480 mg Q3W or Q4W, and the anti-PD-L1 mAb is administered 1200 mg Q3W or Q4W.

[0012] In one embodiment, the anti-fucosyl GM1 mAb, e.g., BMS-986012, is administered at 400 mg or 1000 mg, and the anti-PD-1 mAb, e.g., nivolumab, is administered at 360 mg, both administered every three weeks (Q3W). In another embodiment, the anti-fucosyl GM1 mAb, e.g., BMS-986012, is administered at 400 mg or 1000 mg, and the anti-PD-1 mAb, e.g., nivolumab, is administered at 480 mg, both administered every four weeks (Q4W). In a further embodiment, the anti-fucosyl GM1 antibody and the anti-PD-1 antibody may be co-formulated in the same vial for combined administration.

[0013] In various embodiments, the method includes one, two, three or four treatments, or continues as long as clinical benefit is observed, or until unmanageable toxicity or disease progression occurs. In one embodiment, one or both of the antibodies are formulated for intravenous administration. The efficacy of the treatment methods provided herein can be evaluated using any suitable means, such as reduction in cancer size, reduction in the number of metastatic lesions over time, stable disease, partial response and complete response.

[0014] In some embodiments, the subject with SCLC has not been previously treated with a checkpoint inhibitor. In one embodiment, the subject has previously received a first anti-cancer drug treatment. In another embodiment, the lung cancer is advanced, metastatic, recurrent, and / or refractory lung cancer. In a further embodiment, the subject with SCLC has extensive stage small cell lung cancer (ES-SCLC). In some embodiments, the method of the invention is a first line treatment for lung cancer, such as SCLC. In other embodiments, the method of the invention is a second line treatment for lung cancer, such as SCLC.

[0015] Other features and advantages of the present invention will become apparent from the following detailed description and examples, which should not be construed as limiting. The contents of all references cited throughout this application, including scientific articles, GenBank entries, patents, and patent applications, are hereby expressly incorporated by reference in their entirety as part of this specification. [Brief description of the drawings]

[0016] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] Figure 1 shows tumor growth in a mouse DMS79 tumor model as a function of treatment with anti-fucosyl GM1 BMS-986012 and / or cisplatin. For each data point, the median tumor volume is shown for a group of 8 mice. See Example 1.

[0017] [Diagram 2] Figure 2 shows tumor growth in a mouse DMS79 tumor model as a function of treatment with anti-fucosyl GM1 BMS-986012 and / or etoposide. For each data point, the median tumor volume is shown for a group of 8 mice. See Example 2.

[0018] [Figure 3A] Figures 3A and 3B show exemplary combination therapies of the present invention. Figure 3A provides detailed dosage regimens for induction and maintenance cycles (Arm A) and the same cycles without treatment with anti-fucosyl-GM1 (Arm B), as detailed in Example 5. [Figure 3B] FIG. 3B provides a table illustrating the dosing schedule for the cycle outlined in Example 5 and shown in FIG. 3A. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Detailed Description of the Invention BMS-986012 is a first-in-class fully human monoclonal antibody (mAb) that specifically binds to fucosyl-GM1 ganglioside. BMS-986012 exhibits high affinity and dose-dependent saturable binding to fucosyl-GM1, and no detectable, antigen-specific binding to the closely related molecule GM1. Because fucosyl-GM1 is preferentially found on the surface of lung cancer cells, BMS-986012 is particularly well suited for the treatment of lung cancer, such as small cell lung cancer (SCLC).

[0020] BMS-986012 is non-fucosylated (lacking fucosylation on the Fc domain). The absence of fucosyl groups in BMS-986012 confers higher affinity to Fc receptors, resulting in enhanced antibody-dependent cellular cytotoxicity (ADCC). Furthermore, this antibody has been shown to mediate potent complement-dependent cytotoxicity (CDC) and antibody-dependent cellular phagocytosis (ADCP). See, e.g., WO2007 / 067992, expressly incorporated by reference in its entirety as part of this specification.

[0021] Although BMS-986012 is effective in treating lung cancer as a monotherapy, improved treatment methods are constantly desired.

[0022] Programmed cell death 1 (PD-1) is a cell surface signaling receptor that plays a key role in controlling T cell activation and tolerance. Keir et al.(2008)Ann.Rev.Immunol.26:677-704. It is a type I transmembrane protein that, together with BTLA, CTLA-4, ICOS and CD28, constitutes the CD28 family of T cell costimulatory receptors. PD-1 is expressed primarily on activated T cells, B cells and myeloid cells. Dong et al.(1999)Nat.Med.5:1365-1369. It is also expressed on natural killer (NK) cells. Terme M et al.(2011)Cancer Res.71:5393-5399. Binding of PD-1 by its ligands, PD-L1 and PD-L2, results in phosphorylation of tyrosine residues in the intracellular proximal immunoreceptor tyrosine inhibitory domain, followed by recruitment of the phosphatase SHP-2, and ultimately downregulation of T cell activation. One of the key roles of PD-1 is to limit the activity of T cells in peripheral tissues during inflammatory responses to infection and thus the development of autoimmunity. Pardoll (2012) Nat. Rev. Cancer 12:252-264. Evidence for this negative regulatory role comes from the discovery that PD-1-deficient mice develop lupus-like autoimmune disease, including arthritis and nephritis along with cardiomyopathy. Nishimura et al. (1999) Immunity; 11:141-151; and Nishimura et al. (2001) Science 291:319-322. In the tumor milieu, the result is the development of immune resistance in the tumor microenvironment. PD-1 is highly expressed on tumor-infiltrating lymphocytes, and its ligand is upregulated on the cell surface of many different tumors. Dong et al. (2002) Nat. Med. 8:793-800. Binding of ligands to PD-1 has been demonstrated to result in immune evasion in multiple mouse cancer models. In addition, blocking this interaction results in antitumor activity.Topalian et al.(2012)New Eng.J.Med.366(26):2443-2454;Topalian et al.(2012)Curr.Opin.Immunol.24:207-212;Brahmer et al.(2012)New Eng.J.Med.366(26):2455-2465;Hamid et al. al.(2013)New Eng.J.Med.369:134-144;Hamid & Carvajal(2013)Expert Opin.Biol.a r.13(6):847-861.

[0023] Without intending to be bound by theory, BMS-986012 may mediate the killing of Fucosyl-GM1-expressing lung cancer cells, thereby decreasing the concentration of Fucosyl-GM1 shed into the tumor microenvironment. Early studies have shown that gangliosides released from tumor cells into the tumor microenvironment inhibit tumor-specific immune responses. McKallip et al. (1999) J. Immunol. 163:3718. Such ganglioside-mediated suppression of antitumor immune responses can be mediated, for example, by a shift from IFN-γ production to a more Th2-type T cell response (Crespo et al. (2006) J. Leukocyte. Biol. 79:586) and / or by suppression of dendritic cell maturation (Wolfl et al. (2002) Clin. Exp. Immunol. 130:441; Bennaceur et al. (2006) Int. Immunol. 18:879). Regardless of the detailed mechanism of action, reducing the concentration of gangliosides released by treatment with anti-fucosyl GM1 antibodies is expected to reduce the suppression of this antitumor immune response and, consequently, enhance the efficacy of immunomodulatory agents, such as PD-1 / PD-L1 antagonists, that act by enhancing this response.

[0024] Either or both of the above-mentioned mechanisms of action may occur in the tumor microenvironment. Without intending to be bound by theory, the combination therapy of anti-Fucosyl GM1 antibody and PD-1 antagonist may effectively treat tumors that are not sufficiently immunogenic to generate sufficient anti-tumor immune responses to support monotherapy with PD-1 / PD-L1 antagonist, and that cannot be sufficiently eradicated by monotherapy with anti-Fucosyl GM1 antibody.

[0025] I. Definition So that this disclosure may be more readily understood, certain terms are first defined. As used in this application, unless a different definition is expressly provided herein, each of the following terms shall have the meaning set forth below. Further definitions are set forth throughout the specification.

[0026] As used herein, "BMS-986012" refers to an anti-Fucosyl-GM1 mAb comprising a heavy chain of SEQ ID NO: 3 and a light chain of SEQ ID NO: 4. BMS-986012 also comprises a heavy chain variable domain of SEQ ID NO: 1 and a light chain variable domain of SEQ ID NO: 2. BMS-986012 also comprises the CDR sequences of SEQ ID NO: 5 (CDRH1), SEQ ID NO: 6 (CDRH2), SEQ ID NO: 7 (CDRH3), SEQ ID NO: 8 (CDRL1), SEQ ID NO: 9 (CDRL2), SEQ ID NO: 10 (CDRL3).

[0027] "PD-1 / PD-L1 antagonist" or equivalently "PD-1 antagonist" refers to any agent that inhibits the interaction of PD-1 and PD-L1, such as antagonist antibodies specific for either PD-1 or PD-L1, antibody fragments thereof, soluble receptor constructs, nucleic acid-based inhibitors of PD-1 and / or PD-L1 gene expression or mRNA translation, etc. Such agents include, but are not limited to, nivolumab, pembrolizumab, cemiplimab-rwlc, dostarlimab-gxly, zimberelimab, penpulimab, atezolibumab, durvalumab, avelumab, and envafolimab. Nivolumab, also known as "BMS-936558," refers to an anti-PD-1 mAb comprising a heavy chain of SEQ ID NO: 13 and a light chain of SEQ ID NO: 14. BMS-936558 also comprises a heavy chain variable domain of SEQ ID NO: 11 and a light chain variable domain of SEQ ID NO: 12. BMS-936558 also comprises the CDR sequences of SEQ ID NO: 15 (CDRH1), SEQ ID NO: 16 (CDRH2), SEQ ID NO: 17 (CDRH3), SEQ ID NO: 18 (CDRL1), SEQ ID NO: 19 (CDRL2), SEQ ID NO: 20 (CDRL3). The sequence of pembrolizumab is provided in the sequences claimed in U.S. Patent Nos. 8,354,509 and 8,900,587 (heavy chain CDR1, 2 and 3 are sequences 18, 19 and 20, respectively; light chain CDR1, 2 and 3 are sequences 15, 16 and 17, respectively; the heavy chain sequence is residues 20-446 of sequence 31, and the light chain sequence is residues 20-237 of sequence 36), and CAS Registry No. 1374853-91-4, and in Pembrolizumab: Statement on a Nonproprietary Name Adopted by a USAN Council N13 / 140 (November 27, 2013). The sequence of cemiplimab-rwlc is provided in WHO Drug Information Vol. 32, No. 2 (2018) Proposed INN: List 119 (CAS Registry No. 1801342-60-8).The sequence of dostallimab-gxly is provided in WHO Drug Information Vol.32, No.2(2018) Proposed INN:List 119 (CAS Registry No. 2022215-59-2). The sequence of dimvelelimab is provided in WHO Drug Information Vol.34, No.2(2020) Proposed INN:List 123 (CAS Registry No. 2259860-24-5). The sequence of penprimab is provided in WHO Drug Information Vol.34, No.2(2020) Proposed INN:List 123 (CAS Registry No. 2350298-92-7). Other anti-PD-1 antibodies under regulatory review may also find use in different embodiments of the invention, including retifanlimab (WHO Drug Information Vol. 33, No. 2 (2019) Proposed INN: List 121 (CAS Registry No. 2079108-44-2), sintilimab (WHO Drug Information Vol. 32, No. 2 (2018) Proposed INN: List 119, CAS Registry No. 2072873-06-2), tislelizumab (WHO Drug Information Vol. 31, No. 2 (2017) Proposed INN: List 117, CAS Registry No. 1858168-59-8), toripalimab (WHO Drug Information Vol. 32, No. 2 (2018) Proposed INN: List 117, CAS Registry No. 1858168-59-8), rivaroxaban ... 119, CAS Registry Number 1924598-82-2), geptanolimab (WHO Drug Information Vol. 34, No. 2 (2020) Proposed INN: List 123, CAS Registry Number 2348469-43-0), and serplulimab (WHO Drug Information Vol. 33, No. 2 (2019) Proposed INN: List 121, CAS Registry Number 2231029-82-4).The antibody sequences referred to herein are hereby incorporated by reference as part of the present specification.

[0028] For anti-PD-L1 antibodies, the sequence of atezolibumab is provided in U.S. Pat. No. 8,217,149 (the light chain variable domain sequence is sequence 21 and the heavy chain variable domain sequence is sequence 20 and / or variants thereof containing an additional serine (S) residue between S117 and A118). The sequences of atezolibumab and durvalumab are provided in WHO Drug Information Vol. 29, No. 3 (2015) Recommended INN: List 74. The sequence of avelumab is provided in WHO Drug Information Vol. 30, No. 1 (2016) Recommended INN: List 75. The sequence of embafolimab is provided in WHO Drug Information Vol. 32, No. 4 (2018) Proposed INN: List 120 (CAS Registry Number 2102192-68-5). Other anti-PD-L1 antibodies under regulatory review may also find use in different embodiments of the invention, including, but not limited to, sugemalimab (WHO Drug Information Vol. 33, No. 4 (2019) Proposed INN: List 122, CAS Registry No. 2256084-03-2) and socazolimab (WHO Drug Information Vol. 35, No. 2 (2021) Proposed INN: List 122, CAS Registry No. 2305043-30-3), and anti-PD-L1 antibodies not yet in regulatory review, such as cosibelimab (WHO Drug Information Vol. 33, No. 2 (2019) Proposed INN: List 121 (CAS Registry No. 2216751-26-5), may also be considered. The sequences of the antibodies referenced herein are hereby incorporated by reference as part of this specification.

[0029] Target proteins referenced herein, such as PD-1, are intended to refer to the human orthologues of these proteins (e.g., huPD-1; NP_005009; GeneID 5133), unless otherwise indicated or clear from the context.

[0030] "Administering" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Preferred routes of administration of anti-Fucosyl-GM1 antibodies include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, for example by injection or infusion. As used herein, the term "parenteral administration" refers to a method of administration other than enteral administration and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intrathoracic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, intraarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. Alternatively, non-parenteral routes include topical, epithelial or mucosal routes of administration, for example, intranasal, intravaginal, intrarectal, sublingual or topical administration. Administration can also be, for example, once, multiple times, and / or one or more times over an extended period of time. The interval between doses is referred to as "days" to indicate an interval of approximately 24 hours, although this may vary somewhat due to dosing schedules and other administration delays.

[0031] "Concurrent" administration refers to administration of two different agents, such as an anti-Fucosyl GM1 antibody and an anti-PD-1 or anti-PD-L1 antibody, at the same time or before or after each other, without any intentional delay in administration of one agent. For example, concurrent administration includes simultaneous administration, such as when the agents are co-formulated or mixed prior to administration, and also includes administration of the two agents at a convenient interval, typically during the same visit to a medical facility. Typically, concurrent administration occurs on the same day, excluding administration in separate visits to a medical facility on different days.

[0032] As used herein, the term "adverse event (AE)" refers to an unfavourable and generally unintended or undesirable sign (including abnormal laboratory findings), symptom or disease associated with the use of a medical treatment. A medical treatment may have one or more associated AEs, each of which may have the same or different levels of severity. Reference to a method that can "modify an adverse event" refers to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0033] "Antibody (Ab)" includes, but is not limited to, a glycoprotein immunoglobulin that specifically binds an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or an antigen-binding portion thereof. Each H chain comprises a heavy chain variable region (herein referred to as V H The heavy chain constant region includes three constant domains, C H1 , C H2 and C H3 Each light chain comprises a light chain variable region (referred to herein as V L The light chain constant region comprises one constant domain, C L Includes: V H and V L The regions are further divided into hypervariable regions called complementarity determining regions (CDRs) and interspersed more conserved regions called framework regions (FRs). H and V Lcomprises three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains comprise a binding domain that interacts with an antigen. The constant region of an antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. As used herein, the antibody heavy chain sequence may comprise a C-terminal lysine (K), but this residue may be clipped off in whole or in part during the manufacturing process or removed from the genetic construct used to produce the antibody to avoid potential heterogeneity due to the aforementioned clipping. Both heavy chain sequences provided herein (SEQ ID NOs: 3 and 13) do not comprise a C-terminal lysine residue.

[0034] Immunoglobulins may be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art, and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by the heavy chain constant region. The term "antibody" includes, by way of example, but not limited to, 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 to reduce their immunogenicity in humans.

[0035] An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to Fucosyl-GM1 is substantially free of antibodies that specifically bind to antigens other than Fucosyl-GM1). Furthermore, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0036] The term "monoclonal antibody" ("mAb") refers to a non-naturally occurring preparation of antibody molecules of single molecular composition, i.e., antibody molecules essentially identical in primary sequence, which display a single binding specificity and affinity for a particular epitope. A mAb is an example of an isolated antibody. mAbs may be produced by hybridoma, recombinant, transgenic or other techniques known to those of skill in the art.

[0037] A "human" antibody (HuMAb) refers to an antibody having a variable region in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, when the antibody contains a constant region, the constant region is also derived from a human germline immunoglobulin sequence. The human antibodies of the present invention may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or in vivo somatic mutation). However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human" antibody and "fully human" antibody are used synonymously.

[0038] "Humanized antibody" refers to an antibody in which some, most or all of the amino acids outside the CDR domain of a non-human antibody are replaced with the corresponding amino acids from a human immunoglobulin. In one embodiment of a humanized form of an antibody, some, most or all of the amino acids outside the CDR domain are replaced with amino acids from a human immunoglobulin, while some, most or all of the amino acids within one or more CDR regions remain unchanged. Small additions, deletions, insertions, substitutions or modifications of amino acids are permissible as long as they do not eliminate the binding ability of the antibody to a particular antigen. A "humanized" antibody retains the same antigen specificity as the original antibody.

[0039] "Checkpoint inhibitors" refer to therapeutic agents used to treat cancer that act by inhibiting the activity of "checkpoint" proteins made by several types of immune cells, such as T cells, and some cancer cells. These checkpoint proteins suppress immune responses, for example to prevent unwanted immunopathology, but their action can also prevent immune surveillance that might otherwise eradicate or reduce tumor formation. Checkpoint inhibitors include, but are not limited to, therapeutic antagonist antibodies against PD-1, PD-L1, CTLA-4, TIGIT, LAG3, TIM3, VISTA, and BTLA. Qin et al. (2019) Mol. Cancer 18:155.

[0040] A "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, e.g., the variable region is derived from a murine antibody and the constant region is derived from a human antibody.

[0041] An "anti-antigen" antibody refers to an antibody that specifically binds to an antigen. For example, an anti-Fucosyl GM1 antibody specifically binds to Fucosyl GM1.

[0042] An "antigen-binding portion" of an antibody (also called an "antigen-binding fragment") means one or more fragments of an antibody that retain the ability to specifically bind to the same antigen bound by the full-length antibody.

[0043] "Cancer" refers to a broad group of different diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and proliferation results in the formation of malignant tumors that can invade adjacent tissues and metastasize to distant locations in the body through the lymphatic system or bloodstream. The terms "cancer," "tumor," and "neoplasm" are used interchangeably herein.

[0044] A "subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, non-human primates, vertebrates such as sheep, dogs, and rodents such as mice, rats, and guinea pigs. In a preferred embodiment, the subject is a human. The terms "subject" and "patient" are used interchangeably herein.

[0045] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is that amount of any drug that, when used alone or in combination with another therapeutic agent, protects a subject from developing a disease or promotes disease regression as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods of the disease, or prevention of impairment or disability due to the affliction of the disease. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to those of skill in the art, such as assaying the activity of a drug in human subjects in clinical trials, animal model systems predictive of efficacy in humans, or in in vitro assays.

[0046] As an example, an "anti-cancer drug" promotes the regression of cancer in a subject. In a preferred embodiment, a therapeutically effective amount of a drug promotes the regression of cancer to the point of eliminating the cancer. "Promoting the regression of cancer" means that an effective amount of a drug, administered alone or in combination with an anti-tumor drug, results in a reduction in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free periods, or prevention of functional impairment or disability due to disease affliction. In addition, the terms "effective" and "effectiveness" in relation to treatment include both pharmacological effectiveness and physiological safety. Pharmacological effectiveness refers to the ability of a drug to promote the regression of cancer in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (side effects) at the cell, organ and / or organism level resulting from the administration of a drug.

[0047] As an example of tumor treatment, a therapeutically effective amount of an anticancer drug can preferably inhibit cell proliferation or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80% compared to untreated subjects. In other preferred embodiments of the present invention, tumor regression can be observed, which lasts for at least about 20 days, more preferably at least about 40 days, or even more preferably at least about 60 days. Apart from these final measurements of therapeutic efficacy, the evaluation of immunotherapeutic drugs must also take into account "immune-related" response patterns.

[0048] A therapeutically effective amount of a drug includes a "prophylactically effective amount", which is the amount of drug that inhibits the occurrence or recurrence of cancer when administered alone or in combination with an anti-tumor agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or a subject at risk of recurring cancer. In a preferred embodiment, a prophylactically effective amount completely prevents the occurrence or recurrence of cancer. "Inhibiting" the occurrence or recurrence of cancer means either reducing the likelihood of the occurrence or recurrence of cancer or completely preventing the occurrence or recurrence of cancer.

[0049] The use of alternatives (e.g., "or") should be understood to mean either one, both, or a combination thereof of the options. As used herein, the indefinite article "a" or "an" should be understood to mean "one or more" of any described or listed members.

[0050] The term "about" or "approximately" refers to a value or composition that is within an acceptable error range of a particular value or composition as determined by one of ordinary skill in the art, which may depend in part on how the value or composition is measured or determined, i.e., on the limitations of the measurement system. For example, "about" or "approximately" may mean within 1 or more than 1 standard deviation, as is customary in the art. Alternatively, "about" or "approximately" may mean a range of up to 20%. Furthermore, particularly with respect to biological systems or processes, these terms may mean up to an order of magnitude or up to 5 times a value. When a particular value or composition is described in this application and claims, unless otherwise specified, the meaning of "about" or "approximately" should be considered to be within an acceptable error range of the particular value or composition.

[0051] As described herein, any concentration range, percentage range, ratio range, or integer range, unless otherwise stated, should be understood to include all integer values ​​within the recited range, and, where appropriate, fractions thereof (e.g., tenths and hundredths of an integer). Such ranges further include the values ​​as the limits of the range.

[0052] Various aspects of the invention are described in further detail in the following subsections.

[0053] II. Anti-Fucosyl GM1 Antibody Human monoclonal antibodies that specifically bind to Fucosyl-GM1 with high affinity are disclosed in U.S. Pat. No. 8,383,118 and WO 2007 / 067992 (e.g., human monoclonal antibodies 5B1, 5B1a, 7D4, 7E4, 13B8, and 18D5). Each of the human antibodies disclosed in U.S. Pat. No. 8,383,118 is known to exhibit one or more of the following desirable functional properties: (1) specifically bind to Fucosyl-GM1; (2) bind to Fucosyl-GM1 with high affinity (e.g., 1×10 -7 K under M D(c) binds to the human small cell lung cancer line DMS-79 (human SCLC ATCC#CRL-2049); and (d) inhibits tumor cell proliferation in vitro or in vivo. Preferably, the antibody binds to 5x10 -8 K under M D Binds to Fucosyl-GM1 at 1x10 -8 K under M D Binds to Fucosyl-GM1 at 5x10 -9 K under M D or 1x10 -8 M to 1x10 -10 K less than M D The antibody binds to Fucosyl GM1 at 100-200° C. Standard assays for evaluating the ability of antibody to bind to Fucosyl GM1 are known in the art, including, for example, ELISA, Western blot and RIA. The binding kinetics (e.g., binding affinity) of antibody can also be evaluated by standard assays known in the art, such as ELISA, Scatchard and Biacore analysis.

[0054] A preferred anti-fucosyl GM1 antibody is BMS-986012 (also called MDX-1110 or 7E4). Anti-fucosyl GM1 antibodies that can be used in the disclosed methods also include isolated antibodies that specifically bind to fucosyl GM1 and cross-compete with BMS-986012 for binding to fucosyl GM1 (see, for example, U.S. Patent 8,383,118; WO 2007 / 067992). The ability of antibodies to cross-compete for binding to an antigen suggests that these antibodies bind to the same epitope region of the antigen and sterically hinder the binding to that particular epitope region by other cross-competing antibodies. These cross-competing antibodies are expected to have functional properties very similar to those of BMS-986012 due to their binding to the same epitope region of fucosyl GM1. Cross-competing antibodies can be readily identified in standard Fucosyl-GM1 binding assays, such as Biacore analysis, ELISA assays, or flow cytometry, based on their ability to cross-compete with BMS-986012 (see, for example, WO 2013 / 173223).

[0055] For administration to human subjects, these antibodies are preferably chimeric or, more preferably, humanized or human. Such chimeric, humanized or human monoclonal antibodies can be prepared and isolated by techniques well known in the art. In some, but not all, embodiments, the anti-Fucosyl-GM1 antibodies that can be used in the disclosed inventive methods also comprise antigen-binding portions of the above antibodies. It is well established that the antigen-binding function of an antibody can also be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include (i) Fab fragments, V L , V H , C L and C H1 (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a V H and C H1 (iv) an Fd fragment consisting of the V domain of one arm of an antibody; L and V H Anti-Fucosyl GM1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention may be generated using techniques well known in the art. In other embodiments, such as those in which effector functions are important for activity, antibody fragments will not be suitable for use in the methods of the present invention.

[0056] An exemplary anti-Fucosyl GM1 antibody is BMS-986012, which comprises heavy and light chains comprising the sequences set forth in SEQ ID NOs: 3 and 4, respectively.

[0057] In other embodiments, the antibody has the heavy and light chain CDRs or variable regions of BMS-986012. Thus, in one embodiment, the antibody comprises the VH CDR1, CDR2, and CDR3 domains of BMS-986012 having the sequence set forth in SEQ ID NO:1, and the VL CDR1, CDR2, and CDR3 domains of BMS-986012 having the sequence set forth in SEQ ID NO:2. In another embodiment, the antibody comprises the heavy chain CDR1, CDR2, and CDR3 domains having the sequences set forth in SEQ ID NO:5, 6, and 7, respectively, and the light chain CDR1, CDR2, and CDR3 domains having the sequences set forth in SEQ ID NO:8, 9, and 10, respectively. In another embodiment, the antibody comprises the VH and VL regions having the amino acid sequences set forth in SEQ ID NO:1 and 2, respectively. In another embodiment, the antibody competes for binding with the above-mentioned antibodies and / or binds to the same epitope on Fucosyl-GM1 as the above-mentioned antibodies. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity to the above-mentioned antibodies (e.g., at least about 90%, 95% or 99% variable region identity to SEQ ID NO: 1 or 2).

[0058] III. PD-1 and PD-L1 Antagonists as Immunomodulators PD-1 antagonists suitable for use in the methods described herein include, but are not limited to, ligands, antibodies (e.g., monoclonal and bispecific antibodies), and multivalent agents. In one embodiment, the PD-1 antagonist is a fusion protein, e.g., an Fc fusion protein such as AMP-244. In one embodiment, the PD-1 antagonist is an anti-PD-1 antibody or an anti-PD-L1 antibody. See Twomey & Zhang (2021) The AAPS Journal 23:39.

[0059] An exemplary anti-PD-1 antibody is OPDIVO (登録商標) / Nivolumab (BMS-936558) or an antibody that comprises the CDRs or variable regions of one of antibodies 17D8, 2D3, 4H1, 5C4, 7D3, 5F4, and 4A11 described in WO 2006 / 121168. The sequences for Nivolumab are set forth in SEQ ID NOs: 11-21. In certain other embodiments, the anti-PD-1 antibody is MK-3475 (KEYTRUDA), described in WO 2012 / 145493 and claimed in U.S. Patents 8,354,509 and 8,900,587. (登録商標) / pembrolizumab / formerly lambrolizumab). Cemiplimab-rwlc (the sequence of which can be found in WHO Drug Information Vol.32, No.2 (2018) Proposed INN:List 119 (CAS Registry Number 1801342-60-8)) may also be used. Anti-PD-1 antibodies that compete for binding with one of these antibodies and / or bind to the same epitope on PD-1 may also be used in the combination treatment of the invention.

[0060] Anti-PD-L1 antibodies may also be used in the same embodiment of the invention. Atezolibumab, and durvalumab (whose sequence can be found in WHO Drug Information Vol.29, No.3 (2015) Recommended INN: List 74), and avelumab (whose sequence can be found in WHO Drug Information Vol.30, No.1 (2016) Recommended INN: List 75) may also be used. Anti-PD-L1 antibodies that compete for binding with and / or bind to the same epitope as any of these antibodies may also be used in the combination treatment of the invention.

[0061] The ability of antibodies to cross-compete for binding to an antigen suggests that these antibodies bind to the same epitope region of the antigen and sterically hinder the binding to that particular epitope region by other cross-competing antibodies. These cross-competing antibodies are expected to have functional properties very similar to those of the above anti-PD-1 and anti-PD-L1 antibodies due to their binding to the same epitope region of PD-1 and PD-L1, respectively. Cross-competing antibodies can be easily identified in standard binding assays, such as Biacore analysis, ELISA assays, or flow cytometry, based on their cross-competing ability with the above anti-PD-1 and anti-PD-L1 antibodies.

[0062] For administration to human subjects, these antibodies are preferably chimeric, or more preferably humanized or human. Such chimeric, humanized, and human monoclonal antibodies may be prepared and isolated by techniques well known in the art. In some, but not all, embodiments, the anti-PD-1 and anti-PD-L1 antibodies that can be used in the methods of the disclosed invention also comprise antigen-binding portions of the above antibodies. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include (i) Fab fragments, V and V fragments, and (ii) Fab and V fragments. L , V H , C L and C H1 (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a V H and C H1 (iv) an Fd fragment consisting of the V domain of one arm of an antibody; L and V H It includes an Fv fragment consisting of domains.

[0063] IV. Pharmaceutical Compositions The therapeutic agents of the invention (e.g., anti-Fucosyl GM1 antibodies and / or anti-PD-1 or anti-PD-L1 antibodies, or antigen-binding fragments thereof) can be components of compositions, such as pharmaceutical compositions, that include them and a pharma- ceutically acceptable carrier. Pharmaceutical compositions of the invention include both individual antibodies and co-formulations.

[0064] As used herein, "pharmaceutical acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are physiologically compatible. "Pharmaceutically acceptable" refers to approved by a government regulatory agency or listed in the United States Pharmacopeia or another generally recognized pharmacopoeia for use in animals, particularly humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, including, for example, peanut oil, soybean oil, mineral oil, sesame oil, glycerol polyethylene glycol ricinoleate, and the like. Water or aqueous saline, aqueous dextrose, and glycerol solutions can be employed as carriers, particularly for injectable solutions (e.g., containing anti-fucosyl-GM1 antibodies). Preferably, the carrier for the antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion).The pharmaceutical compositions of the present invention may contain one or more pharma- ceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.

[0065] The liquid composition for parenteral administration can be formulated for administration by injection or continuous infusion.The route of administration by injection or infusion includes intravenous, intraperitoneal, intramuscular, intrathecal and subcutaneous.In one embodiment, anti-Fucosyl GM1 antibody is administered intravenously.

[0066] V. Treatment Method Provided herein is a clinical method for treating lung cancer (e.g., small cell lung cancer) in a subject (e.g., a human subject), comprising administering to the subject a therapeutically effective amount of an anti-Fucosyl GM1 antibody and an anti-PD-1 antibody. In one embodiment, the subject has previously received a first anti-cancer drug treatment. In another embodiment, the lung cancer is progressive, metastatic, recurrent, and / or refractory lung cancer.

[0067] In another embodiment, the antibody is administered as a first line of treatment (e.g., initial or first treatment). In another embodiment, the antibody is administered as a second line of treatment (e.g., after initial or first treatment, including after relapse and / or if the first treatment has failed).

[0068] In certain embodiments, the antibody is administered according to at least one of the following dosing regimens: (a) about 400 mg of antibody every 3 weeks; and (e) about 1000 mg of antibody every 3 weeks. In one embodiment, the antibody is administered at a dose between 400 and 1000 mg, including the range endpoints. Whether stated or not, dose ranges described herein are intended to be inclusive, i.e., doses described as range boundaries are included in the stated dose range. Preferably, administration of the antibody induces a sustained clinical response in the subject. Optionally, administration of the antibody is continued as long as a clinical benefit is observed or until unmanageable toxicity or disease progression occurs. The efficacy of the treatment methods provided herein may be assessed using any suitable means. In one embodiment, the treatment results in at least one therapeutic benefit selected from the group consisting of a reduction in the size of the cancer, a reduction in the number of metastatic lesions over time, stable disease, a partial response, and a complete response.

[0069] Patients treated according to the methods disclosed herein preferably experience an improvement in at least one cancer symptom.In one embodiment, the improvement is measured by the reduction in the amount and / or size of measurable tumor lesions.In another embodiment, the lesions can be measured on chest X-rays, or CT or MRI films.In another embodiment, cytology or histology can be used to evaluate responsiveness to treatment.

[0070] In one embodiment, the treated patient shows a complete response (CR), partial response (PR), or stable disease (SD). In another embodiment, the treated patient experiences tumor shrinkage and / or a slowdown in growth rate, i.e., tumor growth inhibition. In another embodiment, unwanted cell proliferation is reduced or inhibited. In yet another embodiment, one or more of the following occurs: the number of cancer cells is reduced; the size of the tumor is reduced; the invasion of cancer cells into peripheral organs can be inhibited, delayed, slowed, or stopped; tumor metastasis can be slowed or inhibited; tumor growth can be inhibited; tumor recurrence can be prevented or delayed; one or more of the symptoms associated with cancer can be alleviated to some extent.

[0071] VI. Kits and Unit Dosage Forms Also provided herein are kits comprising pharmaceutical compositions comprising an anti-Fucosyl GM1 antibody (such as BMS-986012) and an anti-PD-1 antibody (such as BMS-936558), and a pharma- ceutically acceptable carrier, in therapeutically effective amounts adapted for use in the aforementioned methods.

[0072] The kit may also optionally include instructions, e.g., instructions including an administration schedule, to enable a practitioner (e.g., a doctor or nurse) or a patient to administer the composition included in the kit to a patient with cancer (e.g., lung cancer). The kit may also include a syringe.

[0073] Optionally, the kit comprises a plurality of packages of single-dose pharmaceutical compositions, each of which comprises an effective amount of antibody for single administration, according to the method provided above.The instrument or device required for administering pharmaceutical composition can also be included in the kit.For example, the kit can provide one or more pre-filled syringes that contain a certain amount of antibody.

[0074] The following examples are illustrative only and should not be construed as limiting the scope of the disclosure in any way, since numerous variations and equivalents will become apparent to those of skill in the art upon reading this disclosure.

[0075] The contents of all references, GenBank entries, patents and published patent applications cited throughout this application are hereby expressly incorporated by reference in their entirety as part of this application.

[0076] Example 1 Combination Therapy - Antifucosyl GM1 and Cisplatin The combination therapy method of the present invention involving antibodies against Fucosyl-GM1 and cisplatin was tested in the DMS-79 SCLC tumor model in mice (N=9 mice per group). Bepler et al. (1989) Oncogene 4:45; Petten- gill (1980) Cancer 45:906; Petten- gill et al. (1980) Exp. Cell Biol. 48:279. Briefly, DMS79 cells were cultured in RPMI supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 15 mg / L sodium bicarbonate, 4.5 g / L glucose, 10 mM HEPES, and 10 μM NaPyr, and then implanted subcutaneously into the right flank of male C.B17 SCID mice (0.1 mL phosphate-buffered saline (PBS) and 0.1 mL MATRIGEL per flank). (登録商標) 5 million cells in a gelatinous protein mixture). Tumors were 80–155 mm in mean or median volume, estimated by LxWxH / 2 using digital calipers. 3 Once this was reached, mice were randomly assigned to treatment groups (N=8 mice per group).

[0077] Anti-fucosyl GM1 antibody BMS-986012 was administered ip to mice at 0.3 mg / kg on days 7, 10, 13, 17 and 21 after implantation, and cisplatin was administered at 3 mg / kg on days 7, 14, 21 and 28 after implantation. Because fucosyl GM1 is a ganglioside rather than a protein, and is similar in mice to humans, it was not necessary to use a "mouse surrogate" for the mouse studies. Cisplatin was administered in combination with BMS-986012, and as a control, in combination with an isotype control antibody at 3 mg / kg. Additional controls included an isotype control antibody alone, and a vehicle control. The results are shown in Figure 1. Both anti-fucosyl GM1 antibody treatment and cisplatin treatment effectively reduced tumor growth as monotherapy, but the combination was significantly more effective, almost completely halting tumor growth.

[0078] Example 2 Combination Therapy - Antifucosyl GM1 and Etoposide The combination therapy method of the present invention involving antibodies against Fucosyl GM1 and etoposide was tested in a DMS-79 SCLC tumor model in mice (N=9 mice per group) substantially as described in Example 1. Anti-Fucosyl GM1 antibody BMS-986012 was administered ip to mice at 3 mg / kg on days 7, 11, 15, 18 and 21 after implantation, and etoposide was administered ip at 15 mg / kg on days 7, 9 and 11 after implantation. Etoposide was administered in combination with BMS-986012 and, as a control, in combination with an isotype control antibody at 3 mg / kg. Additional controls included an isotype control antibody alone and a vehicle control. The results are shown in Figure 2. Both anti-Fucosyl GM1 antibody treatment and etoposide treatment reduced tumor growth somewhat effectively as monotherapy, but the combination was more effective.

[0079] Example 3 Treatment of human subjects with anti-Fucosyl-GM1 and etoposide or cisplatin Human subjects were treated and safety evaluated in a Phase 1 / 2 trial (NCT02815592) with the combination of anti-Fucosyl-GM1 mAb BMS-986012 and cisplatin (or carboplatin) and etoposide, essentially as described below: Previously untreated patients (n=14) with extensive-stage small cell lung cancer (ES-SCLC) received 400 mg (n=12) or 1000 mg (n=2) of BMS-986012 on day 1 and 80 mg / m cisplatin on day 1. 2 Patients were treated with either 100 mg / m 2 natriuretic peptide-1 (part 1) or carboplatin with an area under the curve (AUC) of 5 (part 2) administered intravenously, plus etoposide 100 mg / m 2 on days 1, 2, and 3. 2 Treatment consisted of four 21-day cycles of treatment with both parts, followed by maintenance monotherapy with BMS-986012 at 400 mg or 1000 mg Q3W.

[0080] Of the 14 patients who received BMS-986012 in combination with platinum / etoposide, 11 continued the monotherapy period. Three patients did not continue the monotherapy period due to disease progression (n=2) and acute coronary syndrome unrelated to study treatment (n=1). The median age of patients was 62 years (range 49-81 years), of which 79% were male. The combination of BMS-986012 and platinum / etoposide was well tolerated, with most treatment-related adverse events (TRAEs) being grade 1-2. The most common TRAE (all grades; grade ≥3) was pruritus (86%; 7%). In most cases, pruritus resolved with antihistamines or low-dose corticosteroids. Urticaria (7%;7%), neutropenia (7%;7%), xeroderma (7%;0%), conjunctivitis (7%;0%), infusion-related reactions (7%;0%) and dizziness (7%;0%) were also observed. No serious TRAEs or dose-limiting toxicities were reported. The safety profiles of BMS-986012 in combination with cisplatin / etoposide (n=7) and BMS-986012 in combination with carboplatin / etoposide (n=7) in this study were not notably different and, with the exception of clinically manageable pruritus, were similar to those observed historically with platinum / etoposide chemotherapy alone.

[0081] Example 4 Treatment of Human Subjects with Anti-Fucosyl-GM1 and Nivolumab Human subjects were treated with the combination of anti-fucosyl-GM1 mAb BMS-986012 and anti-PD-1 mAb nivolumab in a phase 1 / 2 study (NCT02247349) to evaluate safety and efficacy, substantially as described below. Patients (n=29) with recurrent / refractory small cell lung cancer (SCLC) who had not previously received checkpoint inhibitor (CPI) therapy were treated with 400 mg (n=21) or 1000 mg (n=8) BMS-986012 and 360 mg nivolumab intravenously Q3W with a median follow-up of 18.6 months (range 0.6-41.1 months). The median age of patients was 65 years (range 46-79 years), 52% were male; ECOG performance status was 0 (38%) or 1 (62%). All patients were current (24%) or former smokers (76%). More patients had platinum-sensitive (65%) than platinum-refractory disease (30%).

[0082] The combination was well tolerated with manageable side effects. The most common treatment-related adverse events, all grade / grade ≥3, were pruritus (93% / 21%), fatigue (28% / 0%), dry skin (28% / 0%), and hypothyroidism (17% / 0%). In most patients, pruritus decreased over time and was managed with antihistamines and low-dose corticosteroids. The confirmed ORR with the combination of BMS-986012 and nivolumab was 38% (CR, n=1 [3%]; PR, n=10 [35%]), with an additional 3 patients (10%) showing SD, for an overall disease control rate of 48%. This compares with published ORR of 12% and disease control rate of 29% for nivolumab monotherapy. Ready et al. (2020) J. Thorac. Oncol. 15:426 (147 patients).

[0083] At data cutoff, mDOR was 26.4 months (95% CI, 4.4 months-NR) with 4 patients still on treatment. mPFS was 2.1 months (95% CI, 1.4-9.9 months), and mOS was 18.7 months (95% CI, 8.2 months-NR). No differences in response rates were observed between platinum-sensitive and -refractory populations.

[0084] The results presented here suggest that the combination of BMS-986012 and nivolumab is a promising therapeutic combination for the treatment of patients with recurrent / refractory SCLC who have not been previously treated with CPI therapy, regardless of whether their cancer is platinum-sensitive or refractory. The safety profile was manageable, and although based on a small number of patients, the responses were clinically meaningful and durable.

[0085] Example 5 Combination therapy with carboplatin, etoposide, anti-fucosyl-GM1 and nivolumab Patients with SCLC or ES-SCLC may be treated substantially as described below and as shown in FIG. 3A. Patients are treated with carboplatin at AUC 5 mg / ml / min, 100 mg / ml 2 Patients will be treated over four 21-day rounds of induction therapy that includes 400 mg of etoposide, 420 mg of the anti-fucosyl-GM1 mAb BMS-986012, and 360 mg of the anti-PD-1 mAb nivolumab, all administered iv on day 1 of each cycle. Etoposide will also be administered at the same doses on days 2 and 3 of each cycle.

[0086] After four cycles of induction therapy, patients will be treated with one or more 28-day rounds of maintenance therapy comprising 560 mg of BMS-986012 and 480 mg of the anti-PD-1 mAb nivolumab, all administered iv on day 1 of each cycle.

[0087] [Table 1]

[0088] The antibody sequences in the sequence listing include the sequences of the mature heavy and light chain variable regions, i.e. the sequences do not include the signal peptide.

[0089] Equivalents: Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments disclosed herein. Such equivalents are intended to be encompassed by the scope of the following claims.

Claims

1. A pharmaceutical composition comprising carboplatin, etoposide, an anti-fucosyl GM1 antibody, and an anti-PD-1 antibody or an anti-PD-L1 antibody for use in a method for treating small cell lung cancer (SCLC) in a subject, comprising: the method comprises one or more rounds of induction therapy and one or more rounds of maintenance therapy; a. each round of induction therapy includes treatment with carboplatin, etoposide, said anti-fucosyl-GM1 antibody, and said anti-PD-1 antibody or anti-PD-L1 antibody on day 1; and b. A pharmaceutical composition, wherein each round of maintenance therapy comprises treatment with said anti-Fucosyl GM1 antibody and said anti-PD-1 antibody on day 1.

2. The pharmaceutical composition described in claim 1, wherein the SCSL is extensive-stage small cell lung cancer (ES-SCLC).

3. 3. The pharmaceutical composition of claim 1 or 2, wherein each round of induction therapy is for 21 days (Q3W) and each round of maintenance therapy is for 28 days (Q4W).

4. The anti-Fucosyl GM1 antibody is selected from the group consisting of: a. a heavy chain, i. CDRH1 comprising the sequence of SEQ ID NO:5; ii. a CDRH2 comprising the sequence of SEQ ID NO:6; and iii. a heavy chain comprising a CDRH3 comprising the sequence of SEQ ID NO:7; and b. A light chain, i. CDRL1 comprising the sequence of SEQ ID NO:8; ii. a CDRL2 comprising the sequence of SEQ ID NO:9; and iii. A light chain comprising a CDRL3 comprising the sequence of SEQ ID NO:

10.

5. The anti-Fucosyl GM1 antibody is selected from the group consisting of: a. a heavy chain variable region comprising the sequence of SEQ ID NO:1; and b. a light chain variable region comprising the sequence of SEQ ID NO:

2.

6. The anti-Fucosyl GM1 antibody is selected from the group consisting of: a. a heavy chain comprising the sequence of SEQ ID NO:3; and b. a light chain comprising the sequence of SEQ ID NO: 4, wherein the anti-fucosyl GM1 antibody is non-fucosylated.

7. The anti-PD-1 antibody or the anti-PD-L1 antibody, a. a heavy chain comprising the sequence of SEQ ID NO: 13, and b. The pharmaceutical composition of any one of claims 1 to 6, which is an anti-PD-1 antibody comprising a light chain comprising the sequence of SEQ ID NO:

14.

8. Anti-fucosyl GM1 antibody is administered at 420 mg for induction therapy, and / or The pharmaceutical composition according to claim 6 or 7, wherein the anti-fucosyl GM1 antibody is administered at 560 mg for maintenance therapy.

9. The anti-PD-1 antibody is administered at 360 mg for induction therapy, and / or The pharmaceutical composition of claim 7 or 8, wherein the anti-PD-1 antibody is administered at 480 mg for maintenance therapy.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the method comprises exactly 4 rounds of induction therapy.

11. a. On Day 1 of each Round: i) Carboplatin with AUC 5 mg / ml / min; ii) 100mg / m ２ of etoposide; iii) 420 mg of BMS-986012; and iv) 360 mg nivolumab; four 21-day rounds of induction therapy, including IV administration of b. 100 mg / m on days 2 and 3 of each of four 21-day induction rounds ２ of etoposide, and c. On day 1 of each round of maintenance therapy: i) 560 mg of BMS-986012; and ii) 480 mg nivolumab; The pharmaceutical composition according to any one of claims 1 to 10, comprising one or more rounds of 28 day maintenance therapy comprising iv administration of 12. A pharmaceutical composition comprising an anti-fucosyl GM1 antibody and an immunomodulator selected from an anti-PD-1 antibody and an anti-PD-L1 antibody for use in a method for treating small cell lung cancer (SCLC) in a subject, comprising: The method further comprises administering to the subject a therapeutically effective combination of: a. the anti-fucosyl GM1 antibody administered in a dose of 400 mg or 1000 mg; and b. i. the anti-PD-1 antibody administered in a dose of 360 mg or 480 mg; or ii. the anti-PD-L1 antibody administered at a dose of 1200 mg; A pharmaceutical composition comprising administering

13. The pharmaceutical composition described in claim 12, wherein the SCLC is extensive-stage small cell lung cancer (ES-SCLC).

14. The anti-Fucosyl GM1 antibody is selected from the group consisting of: a. a heavy chain comprising the sequence of SEQ ID NO:3; and b. a light chain comprising the sequence of SEQ ID NO: 4, wherein the anti-fucosyl GM1 antibody is non-fucosylated.

15. An immunomodulatory agent comprising: a. a heavy chain comprising the sequence of SEQ ID NO: 13; and b. The pharmaceutical composition of any one of claims 12 to 14, which is an anti-PD-1 antibody comprising a light chain comprising the sequence of SEQ ID NO:

14.

16. The pharmaceutical composition according to any one of claims 12 to 15, wherein the anti-fucosyl GM1 antibody and the anti-PD-1 antibody are administered on the same day, Q3W or Q4W.

17. 17. The pharmaceutical composition of claim 16, wherein the anti-fucosyl GM1 antibody is BMS-986012 and the anti-PD-1 antibody is nivolumab, further wherein BMS-986012 is administered at 400 mg or 1000 mg and nivolumab is administered at 480 mg, and both are administered on the same day Q4W.

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