EP4673225A2 - Polythérapie triple avec des anticorps anti-pvrig, des anticorps anti-tigit et du pembrolizumab - Google Patents

Polythérapie triple avec des anticorps anti-pvrig, des anticorps anti-tigit et du pembrolizumab

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Publication number
EP4673225A2
EP4673225A2 EP24714367.0A EP24714367A EP4673225A2 EP 4673225 A2 EP4673225 A2 EP 4673225A2 EP 24714367 A EP24714367 A EP 24714367A EP 4673225 A2 EP4673225 A2 EP 4673225A2
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EP
European Patent Office
Prior art keywords
seq
cancer
antibody
light chain
heavy chain
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Pending
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EP24714367.0A
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German (de)
English (en)
Inventor
Adeboye Henry ADEWOYE
Pierre FERRE
Eran Ophir
Gad S. Cojocaru
Inbal BARBIRO
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Compugen Ltd
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Compugen Ltd
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Publication of EP4673225A2 publication Critical patent/EP4673225A2/fr
Pending legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39591—Stabilisation, fragmentation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A61K2039/507—Comprising a combination of two or more separate antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • PVRIG is a transmembrane domain protein of 326 amino acids in length, with a signal peptide (spanning from amino acid 1 to 40), an extracellular domain (spanning from amino acid 41 to 171), a transmembrane domain (spanning from amino acid 172 to 190) and a cytoplasmic domain (spanning from amino acid 191 to 326).
  • the full length human PVRIG protein is shown in Figure 1. There are two methionines that can be start codons, but the mature proteins are identical.
  • the PVRIG proteins contain an immunoglobulin (Ig) domain within the extracellular domain, which is a PVR-like Ig fold domain.
  • the PVR-like Ig fold domain may be responsible for functional counterpart binding, by analogy to the other B7 family members.
  • the PVR-like Ig fold domain of the extracellular domain includes one disulfide bond formed between intra domain cysteine residues, as is typical for this fold and may be important for 1 DB2/ 47495312.3 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO structure-function. These cysteines are located at residues 22 and 93 (or 94).
  • a soluble fragment of PVRIG that can be used in testing of PVRIG antibodies. Included within the definition of PVRIG proteins are PVRIG ECD fragments, including know ECD fragments such as those described in U.S. Patent No.9,714, 289.
  • Anti-PVRIG antibodies (including antigen-binding fragments) that both bind to PVRIG and prevent activation by PVRL2 (e.g., most commonly by blocking the interaction of PVRIG and PVLR2), are used to enhance T cell and/or NK cell activation and be used in treating diseases such as cancer and pathogen infection. As such, formulations for administering such antibodies are needed.
  • TIGIT is a coinhibitory receptor that is highly expressed on effector & regulatory (Treg) CD4+ T cells, effector CD8+ T cells, and NK cells.
  • TIGIT has been shown to attenuate immune response by (1) direct signaling, (2) inducing ligand signaling, and (3) competition with and disruption of signaling by the costimulatory receptor CD226 (also known as DNAM-1).
  • TIGIT signaling has been the most well-studied in NK cells, where it has been demonstrated that engagement with its cognate ligand, poliovirus receptor (PVR, also known as CD155) directly suppresses NK cell cytotoxicity through its cytoplasmic ITIM domain.
  • PVR poliovirus receptor
  • Knockout of the TIGIT gene or antibody blockade of the TIGIT/PVR interaction has shown to enhance NK cell killing in vitro, as well as to exacerbate autoimmune diseases in vivo.
  • TIGIT can induce PVR-mediated signaling in dendritic or tumor cells, leading to the increase in production of anti- inflammatory cytokines such as IL10.
  • TIGIT can also inhibit lymphocyte responses by disrupting homodimerization of the costimulatory receptor CD226, and by competing with it for binding to PVR.
  • TIGIT is highly expressed on lymphocytes, including Tumor Infiltrating Lymphocytes (TILs) and Tregs, that infiltrate different types of tumors.
  • TILs Tumor Infiltrating Lymphocytes
  • Tregs Tregs
  • PVR is also broadly expressed in tumors, suggesting that the TIGIT-PVR signaling axis may be a dominant immune escape mechanism for cancer.
  • TIGIT expression is tightly correlated with the expression of another important coinhibitory receptor, PD1.
  • TIGIT and PD1 are co- expressed on the TILs of numerous human and murine tumors.
  • PD1 inhibition of T cell responses does not involve competition for ligand binding with a costimulatory receptor.
  • DB2/ 47495312.3 2 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO The immune checkpoint, poliovirus receptor related immunoglobulin domain containing (PVRIG, also known as CD112R) represents a new inhibitory receptor within the TIGIT family of receptors.
  • PVRIG poliovirus receptor related immunoglobulin domain containing
  • PVRIG binds with high affinity to its cognate ligand, poliovirus receptor-related 2 (PVRL2, also known as CD112 or nectin-2) to deliver an inhibitory signal through its ITIM motif within T and NK cells.
  • PVRL2 poliovirus receptor-related 2
  • the affinity of TIGIT to PVR and of PVRIG to PVRL2 is higher than the affinity of CD226 to either PVR or PVRL2, suggesting TIGIT and PVRIG can outcompete PVR and PVRL2 from CD226 and providing an indirect mechanism by which TIGIT and PVRIG can reduce lymphocyte function.
  • TIGIT and PVRIG two receptors with the same family, deliver inhibitory signals to dampen T and NK cell responses.
  • antibodies to both TIGIT and PVRIG are useful for triple therapy combinations pembrolizumab and meet the unmet need of providing triple combination therapies for use in treating patients or subjects refractory or resistant to multiple lines of treatment.
  • BRIEF SUMMARY OF THE INVENTION it is an object of the invention to provide triple combination therapies for methods of treatment or for use in treatment, wherein the triple combination includes stable liquid pharmaceutical formulations comprising anti-PVRIG antibodies, anti-TIGIT antibodies, and pembrolizumab.
  • the anti-PVRIG antibody comprises: i) a heavy chain variable domain comprising a vhCDR1 comprising SEQ ID NO: 5, a vhCDR2 comprising SEQ ID NO: 6, a vhCDR3 comprising SEQ ID NO: 7; and ii) a light chain variable domain comprising a vlCDR1 comprising SEQ ID NO: 10, a vlCDR2 comprising SEQ ID NO: 11, a vlCDR3 comprising SEQ ID NO: 12.
  • the anti-PVRIG antibody comprises: i) a heavy chain variable domain comprising SEQ ID NO:4, and ii) a light chain variable domain comprising SEQ ID NO:9.
  • the anti-PVRIG antibody comprises: i) a heavy chain comprising a vhCDR1 comprising SEQ ID NO: 5, a vhCDR2 comprising SEQ ID NO: 6, a vhCDR3 comprising SEQ ID NO: 7; and ii) a light chain comprising a vlCDR1 comprising SEQ ID NO: 10, a vlCDR2 comprising SEQ ID NO: 11, a vlCDR3 comprising SEQ ID NO: 12; wherein the heavy chain has at least 90% identity to SEQ ID NO: 8, and the light chain has at least 90% identity to SEQ ID NO: 13.
  • the anti-PVRIG antibody comprises: i) a heavy chain variable domain comprising a vhCDR1 comprising SEQ ID NO: 15, a vhCDR2 comprising SEQ ID NO: 16, a vhCDR3 comprising SEQ ID NO: 17; and ii) a light chain variable domain comprising a vlCDR1 comprising SEQ ID NO: 20, a vlCDR2 comprising SEQ ID NO: 21, a vlCDR3 comprising SEQ ID NO: 22.
  • the anti-PVRIG antibody comprises: i) a heavy chain variable domain comprising SEQ ID NO:14, and ii) a light chain variable domain comprising SEQ ID NO:19.
  • the anti-PVRIG antibody comprises: i) a heavy chain comprising a vhCDR1 comprising SEQ ID NO: 15, a vhCDR2 comprising SEQ ID NO: 16, a vhCDR3 comprising SEQ ID NO: 17; and ii) a light chain comprising a vlCDR1 comprising SEQ ID NO: 20, a vlCDR2 comprising SEQ ID NO: 21, a vlCDR3 comprising SEQ ID NO: 22; wherein the heavy chain has at least 90% identity to SEQ ID NO: 18, and the light chain has at least 90% identity to SEQ ID NO: 23.
  • the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising a vhCDR1 comprising SEQ ID NO: 25, a vhCDR2 comprising SEQ ID NO: 26, a vhCDR3 comprising SEQ ID NO: 27; and ii) a light chain variable domain comprising a vlCDR1 comprising SEQ ID NO: 30, a vlCDR2 comprising SEQ ID NO: 31, a vlCDR3 comprising SEQ ID NO: 32.
  • the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising SEQ ID NO:24, and ii) a light chain variable domain comprising SEQ ID NO:29. [0020] In some embodiments, the anti-TIGIT antibody comprises: DB2/ 47495312.3 5 MLB Ref.
  • No.114386-5030-WO Compugen Ref.230-WO i) a heavy chain comprising a vhCDR1 comprising SEQ ID NO: 25, a vhCDR2 comprising SEQ ID NO: 26, a vhCDR3 comprising SEQ ID NO: 27; and ii) a light chain comprising a vlCDR1 comprising SEQ ID NO: 30, a vlCDR2 comprising SEQ ID NO: 31, a vlCDR3 comprising SEQ ID NO: 32; wherein the heavy chain has at least 90% identity to SEQ ID NO: 28, and the light chain has at least 90% identity to SEQ ID NO: 33.
  • the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising a vhCDR1 comprising SEQ ID NO: 35, a vhCDR2 comprising SEQ ID NO: 36, a vhCDR3 comprising SEQ ID NO: 37; and ii) a light chain variable domain comprising a vlCDR1 comprising SEQ ID NO: 40, a vlCDR2 comprising SEQ ID NO: 41, a vlCDR3 comprising SEQ ID NO: 42.
  • the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising SEQ ID NO:34, and ii) a light chain variable domain comprising SEQ ID NO:39.
  • the anti-TIGIT antibody comprises: i) a heavy chain comprising a vhCDR1 comprising SEQ ID NO: 35, a vhCDR2 comprising SEQ ID NO: 36, a vhCDR3 comprising SEQ ID NO: 37; and ii) a light chain comprising a vlCDR1 comprising SEQ ID NO: 40, a vlCDR2 comprising SEQ ID NO: 41, a vlCDR3 comprising SEQ ID NO: 42; wherein the heavy chain has at least 90% identity to SEQ ID NO: 38, and the light chain has at least 90% identity to SEQ ID NO: 43.
  • pembrolizumab comprises: i) a heavy chain variable domain comprising a vhCDR1 comprising SEQ ID NO: 45, a vhCDR2 comprising SEQ ID NO: 46, a vhCDR3 comprising SEQ ID NO: 47; and DB2/ 47495312.3 6 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO ii) a light chain variable domain comprising a vlCDR1 comprising SEQ ID NO: 50, a vlCDR2 comprising SEQ ID NO: 51, a vlCDR3 comprising SEQ ID NO: 52.
  • pembrolizumab comprises: i) a heavy chain variable domain comprising SEQ ID NO: 44, and ii) a light chain variable domain comprising SEQ ID NO: 49.
  • pembrolizumab comprises: i) a heavy chain comprising a vhCDR1 comprising SEQ ID NO: 45, a vhCDR2 comprising SEQ ID NO: 46, a vhCDR3 comprising SEQ ID NO: 47; and ii) a light chain comprising a vlCDR1 comprising SEQ ID NO: 50, a vlCDR2 comprising SEQ ID NO: 51, a vlCDR3 comprising SEQ ID NO: 52; wherein the heavy chain has at least 90% identity to SEQ ID NO: 48, and the light chain has at least 90% identity to SEQ ID NO: 53.
  • the anti-PVRIG antibody is administered as a stable liquid pharmaceutical formulation and, wherein the stable liquid pharmaceutical formulation of the anti-PVRIG antibody comprises: (a) from 10 mM to 100 mM histidine; (b) from 30 mM to 100 mM NaCl; (c) from 20 mM to 150 mM L-Arginine; and (d) from 0.005% to 0.1% w/v polysorbate 80, wherein the formulation has a pH from 5.5 to 7.0.
  • the anti-PVRIG antibody and/or the anti-TIGIT antibody comprises a CH1-hinge-CH2-CH3 sequence of IgG4 (SEQ ID NO:57 or SEQ ID NO:58), wherein the hinge region optionally comprises mutations.
  • the anti-PVRIG antibody and/or the anti-TIGIT antibody comprises the CH1-hinge-CH2-CH3 region from IgG1, IgG2, IgG3, or IgG4, wherein the hinge region optionally comprises mutations.
  • the IgG1 comprises SEQ ID NO:54.
  • the IgG2 comprises SEQ ID NO:55. In some DB2/ 47495312.3 7 MLB Ref.
  • the IgG3 comprises SEQ ID NO:56.
  • the IgG4 comprises SEQ ID NO:57.
  • the IgG4 comprises SEQ ID NO:58.
  • the anti-PVRIG heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and the anti-PVRIG light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9).
  • the anti-PVRIG heavy chain variable domain is from the heavy chain of CHA.7.538.1.2.H4(S241P) (SEQ ID NO:14) and the anti-PVRIG light chain variable domain is from the light chain of CHA.7.538.1.2.H4(S241P) (SEQ ID NO:19).
  • the anti-PVRIG antibody comprises a CL region of human kappa 2 light chain.
  • the pharmaceutical formulation comprises from 10 mM to 80 mM histidine, from 15 mM to 70 mM histidine, from 20 mM to 60 mM histidine, from 20 mM to 50 mM histidine, or from 20 mM to 30 mM histidine. [0034] In some embodiments, the pharmaceutical formulation comprises about 25 mM histidine. [0035] In some embodiments, the pharmaceutical formulation comprises from 30 mM to 100 mM NaCl, from 30 mM to 90 mM NaCl, from 40 mM to 80 mM NaCl, from 30 mM to 70 mM histidine, or from 45 mM to 70 mM NaCl.
  • the pharmaceutical formulation comprises about 60 mM NaCl.
  • the pharmaceutical formulation comprises from 20 mM to 140 mM L-arginine, from 30 mM to 140 mM L-arginine, from 40 mM to 130 mM L-arginine, from 50 mM to 120 mM L-arginine, from 60 mM to 110 mM L-arginine, from 70 mM to 110 mM L-arginine, from 80 mM to 110 mM L-arginine, or from 90 mM to 110 mM L-arginine.
  • the pharmaceutical formulation comprises about 100 mM L- arginine.
  • pharmaceutical formulation comprises from 0.006% to 0.1% w/v polysorbate 80, from 0.007% to 0.09% w/v polysorbate 80, from 0.008% to 0.08% w/v polysorbate 80, from 0.009% to 0.09% w/v polysorbate 80, from 0.01% to 0.08% w/v polysorbate 80, from 0.01% to 0.07% w/v polysorbate 80, from 0.01% to 0.07% w/v polysorbate 80, or from 0.01% to 0.06% w/v polysorbate 80, or from 0.009% to 0.05% w/v polysorbate 80.
  • the pharmaceutical formulation comprises about 0.01% polysorbate 80.
  • the pH is from 6 to 7.0. In some embodiments, the pH is from 6.3 to 6.8. In some embodiments, the pH is 6.5 +/- 0.2.
  • the anti-PVRIG antibody is at a concentration of from 10 mg/mL to 40 mg/mL, 15 mg/mL to 40 mg/mL, 15 mg/mL to 30 mg/mL, 10 mg/mL to 25 mg/mL, or 15 mg/mL to 25 mg/mL.
  • the anti-PVRIG antibody formulation is stable at -20°C for at least 1 month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months, 30 months, 36 months, 42 months or 48 months. [0044] In some embodiments, the anti-PVRIG antibody formulation is stable at 2°C to 8°C for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks, 1 month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months, 30 months, 36 months, 42 months or 48 months.
  • the anti-PVRIG antibody formulation is stable at about 20°C to 25 °C for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks, 1 months, 3 months, 6 months, or 9 months, 12 months, 18 months, or 36 months. [0046] In some embodiments, the anti-PVRIG antibody formulation is stable at 35°C to 40°C for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks.
  • the anti-PVRIG antibody comprises: a) a heavy chain comprising: i) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and b) a light chain comprising: i) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or a) a heavy chain comprising: DB2/ 47495312.3 9 MLB Ref.
  • a VH-CH1-hinge-CH2-CH3 wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH3 region is from IgG4; and b) a light chain comprising: i) a VL-CL, wherein the VL from CHA.7.538.1.H4(S241P) (SEQ ID NO:19) and wherein the CL region is from human kappa 2 light chain.
  • the hinge region optionally comprises mutations.
  • the anti-PVRIG antibody formulation comprises: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.2.H4(S241P) (SEQ ID NO:18); and ii) a light chain comprising the light chain from CHA.7.538.1.2.H4(S241P) (SEQ ID NO:23).
  • the anti-PVRIG antibody formulation comprises: (a) an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises: i) a heavy chain comprising the heavy chain from CHA.7.538.1.2.H4(S241P) (SEQ ID NO:14); and DB2/ 47495312.3 10 MLB Ref.
  • the anti-PVRIG antibody formulation comprising: (a) an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises: i) a heavy chain comprising the heavy chain from CHA.7.538.1.2.H4(S241P) (SEQ ID NO:18); and ii) a light chain comprising the light chain from CHA.7.538.1.2.H4(S241P) (SEQ ID NO:23); (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about
  • the anti-TIGIT antibody comprises: a) a heavy chain comprising VH-CH1-hinge-CH2-CH3; and DB2/ 47495312.3 11 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO b) a light chain comprising VL-VC, wherein VC is either kappa or lambda.
  • the sequence of the CH1-hinge-CH2-CH3 is selected from human IgG1, IgG2 and IgG4, and variants thereof.
  • the hinge region optionally comprises mutations.
  • the anti-TIGIT antibody comprises: i) a heavy chain comprising the heavy chain from CPA.9.083.H4(S241P) (SEQ ID NO:28); and ii) a light chain comprising the light chain from CPA.9.083.H4(S241P) (SEQ ID NO:33); or i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43).
  • the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg of the anti-PVRIG antibody.
  • the anti-PVRIG antibody is administered at a concentration of about 15 mg/kg or about 20 mg/kg.
  • the anti-PVRIG antibody is administered at a concentration of about 20 mg/kg every 3 weeks.
  • the anti-PVRIG antibody is administered at a concentration of about 15 mg/kg every 3 weeks.
  • the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti- TIGIT antibody.
  • the anti-TIGIT antibody is administered about 10 mg/kg every 3 weeks.
  • the anti-TIGIT antibody is administered about 3 mg/kg every 3 weeks.
  • pembrolizumab is administered as a dosage of about 2 mg/kg or 10 mg/kg.
  • pembrolizumab is administered as a dosage of about 2 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 10 mg/kg. [0069] In some embodiments, pembrolizumab is administered as a dosage of about 2 mg/kg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 10 mg/kg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 200 mg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 400 mg every 6 weeks.
  • the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered in any order.
  • the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered simultaneously.
  • DB2/ 47495312.3 13 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO In some embodiments, the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered sequentially.
  • the order of administration is one of: i) anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab; ii) anti-TIGIT antibody, anti-PVRIG antibody, and pembrolizumab; iii) pembrolizumab, anti-PVRIG antibody, and anti-TIGIT antibody; iv) pembrolizumab, anti-TIGIT antibody, and anti-PVRIG antibody; v) anti-PVRIG antibody, pembrolizumab, and anti-TIGIT antibody; or vi) anti-TIGIT antibody, pembrolizumab, and anti-PVRIG antibody.
  • pembrolizumab is administered over about 10 minutes, over about 15 minutes, over about 20 minutes, over about 25 minutes, over about 30 minutes, over about 35 minutes, or over about 40 minutes. [0075] In some embodiments, the pembrolizumab is administered over about 10 minutes to about 40 minutes, over about 15 minutes to about 40 minutes, over about 20 minutes to about 40 minutes, or about 30 minutes +/- 10 minutes [0076] In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 340%, 350%, 360%, 370%, 380%, 390%, 400%, 410%, 420%, 430%, 440%, 45
  • the level of serum IFN ⁇ in the subject is increased by at least 200%, 300%, 400%, or more, relative to untreated subjects. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1000%, 1100%, 1200%, 1300%, 1400%, 1500% or more, relative to untreated subjects. DB2/ 47495312.3 14 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1200%, or more, relative to untreated subjects.
  • the subject exhibits an increased level of serum IFN ⁇ of at least 10 pg/mL, 20 pg/mL, 30 pg/mL, 40 pg/mL, 50 pg/mL, 60 pg/mL, 70 pg/mL, 80 pg/mL, 90 pg/mL, 100 pg/mL, 110 pg/mL, 120 pg/mL, 130 pg/mL, 140 pg/mL, 150 pg/mL, 160 pg/mL, 170 pg/mL, 180 pg/mL, 190 pg/mL, 200 pg/mL, 210 pg/mL, 220 pg/mL, 230 pg/mL, 240 pg/mL, 250 pg/mL, 260 pg/mL, 270 pg/mL, 280 pg/m
  • the subject exhibits an increased level of serum IFN ⁇ of at least 20 pg/mL, 30 pg/mL, 40 pg/mL, or more. In some embodiments, the subject exhibits an increased level of serum IFN ⁇ of at least 40 pg/mL, 50 pg/mL, 60 pg/mL, 70 pg/mL, 80 pg/mL or more. In some embodiments, the subject exhibits an increased level of serum IFN ⁇ of at least 70 pg/mL, or more. In some embodiments, the subject exhibits an increased level of serum IFN ⁇ of at least 80 pg/mL, or more.
  • the cancer is selected from the group consisting of prostate cancer, liver cancer (HCC), rectal cancer, colorectal cancer (CRC), colorectal cancer MSS (MSS-CRC; microsatellite stable colorectal carcinoma/carcinoma), CRC (MSS unknown), metastatic MSS-CRC, refractory metastatic MSS-CRC, MSS-CRC with liver metastasis, ovarian cancer (including ovarian carcinoma), primary peritoneal ovarian carcinoma, advanced epithelial ovarian cancer, advanced epithelial ovarian carcinoma, platinum resistant ovarian cancer, high grade serous adenocarcinoma, endometrial cancer (including endometrial carcinoma), breast cancer, pancreatic cancer, stomach cancer, cervical cancer, head and neck cancer, thyroid cancer, testis cancer, urothelial cancer, lung cancer, melanoma, non-melanoma skin cancer (squamous and basal cell carcinoma), uveal melanoma
  • No.114386-5030-WO Compugen Ref.230-WO atypical carcinoid lung cancer, NSCLC with PDL1 > 50% TPS, cervical SCC, pancreatic cancer, pancreatic adenocarcinoma, adenoid cystic cancer (including adenoid cystic carcinoma), primary peritoneal cancer, microsatellite stable primary peritoneal cancer, platinum resistant microsatellite stable primary peritoneal cancer, Myelodysplastic syndromes (MDS), HNSCC, PD1 refractory or relapsing cancer, gastroesophageal junction cancer, gastric cancer, chordoma, sarcoma, endometrial sarcoma, chondrosarcoma, uterine sarcoma, plasma cell disorders, multiple myeloma, amyloidosis, AL-amyloidosis, glioblastoma, astrocytoma and fallopian tube cancer.
  • MDS Myel
  • the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab combination can find use in a method of treating cancer in a subject.
  • the invention provides for the use of the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab in the manufacture of a medicament for the treatment of cancer in a subject, wherein the anti-PVRIG antibody is formulated as the stable liquid pharmaceutical formulation as described herein.
  • the cancer is selected from the group consisting of prostate cancer, liver cancer (HCC), rectal cancer, colorectal cancer (CRC), colorectal cancer MSS (MSS-CRC; microsatellite stable colorectal carcinoma/carcinoma), CRC (MSS unknown), metastatic MSS-CRC, refractory metastatic MSS-CRC, MSS-CRC with liver metastasis, ovarian cancer (including ovarian carcinoma), primary peritoneal ovarian carcinoma, advanced epithelial ovarian cancer, advanced epithelial ovarian carcinoma, platinum resistant ovarian cancer, high grade serous adenocarcinoma, endometrial cancer (including endometrial carcinoma), breast cancer, pancreatic cancer, stomach cancer, cervical cancer, head and neck cancer, thyroid cancer, testis cancer, urothelial cancer, lung cancer, melanoma, non-melanoma skin cancer (squamous and basal cell carcinoma
  • adenoid cystic cancer including adenoid cystic carcinoma
  • primary peritoneal cancer microsatellite stable primary peritoneal cancer
  • platinum resistant microsatellite stable primary peritoneal cancer platinum resistant microsatellite stable primary peritoneal cancer
  • Myelodysplastic syndromes (MDS) Myelodysplastic syndromes
  • HNSCC Myelodysplastic syndromes
  • PD1 refractory or relapsing cancer gastroesophageal junction cancer, gastric cancer, chordoma, sarcoma, endometrial sarcoma, chondrosarcoma, uterine sarcoma, plasma cell disorders, multiple myeloma, amyloidosis, AL-amyloidosis, glioblastoma, astrocytoma and fallopian tube cancer.
  • the subject is refractory or resistant to treatment with at least two or more prior therapeutic treatments, wherein the prior therapeutic treatments include one or more of fluroropyrimidines (or fluoropyrimidines), irinotecan, oxaliplatin, anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies.
  • fluroropyrimidines or fluoropyrimidines
  • irinotecan irinotecan
  • oxaliplatin anti-PD-1
  • anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody anti-CD137 antibody
  • anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies include one or more of fluroropyrimidines (or fluoropyrimidines), irinotecan, oxaliplatin, anti
  • the subject is refractory or resistant to treatment with at least two or more prior therapeutic treatments, wherein the two or more prior therapeutic treatments include fluroropyrimidines (or fluoropyrimidines), irinotecan, and/or oxaliplatin.
  • the two or more prior therapeutic treatments include fluroropyrimidines (or fluoropyrimidines), irinotecan, and/or oxaliplatin.
  • the subject is refractory or resistant to treatment with no more than three prior therapeutic treatments.
  • the subject is refractory or resistant to treatment with at least three or more prior therapeutic treatments.
  • the subject is refractory or resistant to treatment with at least three or more prior therapeutic treatments, where in the three prior therapeutic treatments include fluroropyrimidines (or fluoropyrimidines), irinotecan, and/or oxaliplatin.
  • the subject has not had prior treatment with an anti-PD-1, anti-PD-L1, or anti-PD-L2- directed therapy.
  • the subject has not had prior treatment with an immune checkpoint inhibitor, DB2/ 47495312.3 17 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO including anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies.
  • an immune checkpoint inhibitor DB2/ 47495312.3 17 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO including anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies.
  • BRIEF DESCRIPTION OF THE DRAWINGS [0089] Figure 1 depicts the full-length sequence of human PVRIG.
  • FIG. 2 depicts the sequence of the human Poliovirus receptor-related 2 protein (PVLR2, also known as nectin-2, CD112 or herpesvirus entry mediator B, (HVEB)), the binding partner of PVRIG.
  • PVLR2 is a human plasma membrane glycoprotein.
  • Figure 3A-3B depicts the variable heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences of the anti-PVRIG antibodies of the invention CHA.7.518.1.H4(S241P) and CHA.7.538.1.2.H4(S241P).
  • Figure 4A-4B depicts the variable heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences of the anti-TIGIT antibodies of the invention CPA.9.083.H4(S241P) and CPA.9.086.H4(S241P).
  • Figure 5 depicts the variable heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences of pembrolizumab.
  • Figure 6 depicts the sequences of human IgG1, IgG2, IgG3 and IgG4.
  • Figure 7 depicts formulation parameters, including buffers and excipients for the CHA.7.518.1.H4(S241P) antibody formulation.
  • Figure 8 depicts an exemplary study design overview.
  • Figure 9 illustrates pharmacodynamic activation of the immune system with CHA.7.518.1.H4(S241P), CPA.9.086.H4(S241P), and Pembrolizumab.
  • C2D1 Peripheral blood was collected from 19 patients (17 CRC (gray) and 2 OVCA (black)) at baseline (C1D1), one day post treatment (C1D2), one week post treatment (C1D8) and before the start of consecutive drug administration (IV, Q3W; every 3 weeks, i.e., C2D1 reflects blood collected 3 weeks post-treatment and prior to the second cycle of treatment) of CHA.7.518.1.H4(S241P) (15 mg/kg ), CPA.9.086.H4(S241P) (3 mg/kg) and Pembrolizumab (200 mg). Samples were assessed for serum IFN ⁇ levels, measured using a pro-inflammatory human cytokine 10-plex assay kit and Meso Scale Discovery (MSD).
  • MSD Meso Scale Discovery
  • Cancer can be considered as an inability of the patient or subject (the terms patient and subject are generally used interchangeably) to recognize and eliminate cancerous cells.
  • these transformed (e.g., cancerous) cells counteract immunosurveillance.
  • Restoring the capacity of immune effector cells—especially T cells— to recognize and eliminate cancer is the goal of immunotherapy.
  • T cell checkpoint inhibitory antibodies such as Yervoy, Keytruda and Opdivo.
  • These antibodies are generally referred to as “checkpoint inhibitors” because they block normally negative regulators of T cell immunity. It is generally understood that a variety of immunomodulatory signals, both costimulatory and coinhibitory, can be used to orchestrate an optimal antigen-specific immune response. Generally, these antibodies bind to checkpoint inhibitor proteins such as CTLA-4 and PD-1, which under normal circumstances prevent or suppress activation of cytotoxic T cells (CTLs).
  • CTLs cytotoxic T cells
  • the present invention is directed to formulations comprising antibodies to human Poliovirus Receptor Related Immunoglobulin Domain Containing Protein, or “PVRIG”, sometimes also referred to herein as “PV protein”.
  • PVRIG Poliovirus Receptor Related Immunoglobulin Domain Containing Protein
  • the present invention provides formulations comprising antibodies, including antigen binding domains, that bind to the human PVRIG and peptides thereof and methods of activating T cells and/or NK cells to treat diseases such as cancer and infectious diseases, and other conditions where increased immune activity results in treatment.
  • formulations comprising antibodies, including antigen binding domains, that bind to the human PVRIG and peptides thereof and methods of activating T cells and/or NK cells to treat diseases such as cancer and infectious diseases, and other conditions where increased immune activity results in treatment.
  • DB2/ 47495312.3 19 MLB Ref.
  • the invention provides formulations comprising antibodies comprising heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences from CHA.7.518.1.H4(S241P) as well as heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences from CHA.7.538.1.H4(S241P).
  • the present invention also provides antibodies, including antigen binding domains, that bind to the human TIGIT protein and peptides thereof.
  • the invention provides formulations comprising antibodies comprising heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences from CPA.9.083.H4(S241P) as well as heavy and light chains as well as the vhCDR1, vhCDR2, vhCDR3, vlCDR1, vlCDR2 and vlCDR3 sequences from CPA.9.086.H4(S241P).
  • PVRIG PROTEINS [0103] The present invention provides formulations comprising antibodies that specifically bind to PVRIG proteins. “Protein” in this context is used interchangeably with “polypeptide”, and includes peptides as well. The present invention provides antibodies that specifically bind to PVRIG proteins.
  • PVRIG is a transmembrane domain protein of 326 amino acids in length, with a signal peptide (spanning from amino acid 1 to 40), an extracellular domain (spanning from amino acid 41 to 171), a transmembrane domain (spanning from amino acid 172 to 190) and a cytoplasmic domain (spanning from amino acid 191 to 326).
  • the full length human PVRIG protein is shown in Figure 1.
  • PVRIG or “PVRIG protein” or “PVRIG polypeptide” may optionally include any such protein, or variants, conjugates, or fragments thereof, including but not limited to known or wild type PVRIG, as described herein, as well as any naturally occurring splice variants, amino acid variants or isoforms, and in particular the ECD fragment of PVRIG.
  • soluble form of PVRIG is also used interchangeably with the terms “soluble ectodomain (ECD)” or “ectodomain” or “extracellular domain (ECD) as well as “fragments of PVRIG polypeptides”, which may refer broadly to one or more of the following optional polypeptides: [0105]
  • the PVRIG proteins contain an immunoglobulin (Ig) domain within the extracellular domain, which is a PVR-like Ig fold domain.
  • the PVR-like Ig fold domain may be responsible for functional counterpart binding, by analogy to the other B7 family members. DB2/ 47495312.3 20 MLB Ref.
  • the PVR-like Ig fold domain of the extracellular domain includes one disulfide bond formed between intra domain cysteine residues, as is typical for this fold and may be important for structure-function. These cysteines are located at residues 22 and 93 (or 94).
  • a soluble fragment of PVRIG that can be used in testing of PVRIG antibodies. Included within the definition of PVRIG proteins are PVRIG ECD fragments, including know ECD fragments such as those described in U.S. Patent No.9,714, 289, incorporate by reference herein in its entirety for all purposes.
  • the anti-PVRIG antibodies (including antigen-binding fragments) that both bind to PVRIG and prevent activation by PVRL2 (e.g. most commonly by blocking the interaction of PVRIG and PVLR2), are used to enhance T cell and/or NK cell activation and be used in treating diseases such as cancer and pathogen infection.
  • PVRL2 e.g. most commonly by blocking the interaction of PVRIG and PVLR2
  • the present invention provides antibodies that specifically bind to TIGIT proteins and prevent activation by its ligand protein, PVR, poliovirus receptor (aka, CD155) a human plasma membrane glycoprotein.
  • TIGIT T cell immunoreceptor with Ig and ITIM domains
  • WUCAM WUCAM
  • Vstm3 Vsig9.
  • TIGIT has an immunoglobulin variable domain, a transmembrane domain, and an immunoreceptor tyrosine-based inhibitory motif (ITIM) and contains signature sequence elements of the PVR protein family.
  • ITIM immunoreceptor tyrosine-based inhibitory motif
  • the extracellular domain (ECD) sequences of TIGIT and of PVR are shown in Figure 1B.
  • TIGIT or “TIGIT protein” or “TIGIT polypeptide” may optionally include any such protein, or variants, conjugates, or fragments thereof, including but not limited to known or wild type TIGIT, as described herein, as well as any naturally occurring splice variants, amino acid variants or isoforms, and in particular the ECD fragment of TIGIT.
  • anti-TIGIT antibodies including antigen-binding fragments that both bind to TIGIT and prevent activation by PVR (e.g., most commonly by blocking the interaction of TIGIT and PVR), are used to enhance T cell and/or NK cell activation and be used in treating diseases such as cancer and pathogen infection.
  • PVR e.g., most commonly by blocking the interaction of TIGIT and PVR
  • the invention provides anti-PVRIG antibodies that can be formulated according to the formulations described herein and which are provided in Figure 3 (e.g., including anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3).
  • PVRIG also called Poliovirus Receptor Related Immunoglobulin Domain Containing Protein, Q6DKI7 or C7orf15, relates to amino acid and nucleic acid sequences shown in RefSeq accession identifier NP_076975, shown in Figure 1.
  • the antibodies of the invention are specific for the PVRIG extracellular domain.
  • the term “antibody” is used generally.
  • Antibodies that find use in the present invention can take on a number of formats as described herein, including traditional antibodies as well as antibody derivatives, fragments and mimetics, described below.
  • the term “antibody” includes any polypeptide that includes at least one antigen binding domain, as more fully described below.
  • Antibodies may be polyclonal, monoclonal, xenogeneic, allogeneic, syngeneic, or modified forms thereof, as described herein, with monoclonal antibodies finding particular use in many embodiments.
  • antibodies of the invention bind specifically or substantially specifically to PVRIG molecules.
  • monoclonal antibodies and “monoclonal antibody composition”, as used herein, refer to a population of antibody molecules that contain only one species of an antigen-binding site capable of immunoreacting with a particular epitope of an antigen
  • polyclonal antibodies and “polyclonal antibody composition” refer to a population of antibody molecules that contain multiple species of antigen-binding sites capable of interacting with a particular antigen.
  • a monoclonal antibody composition typically displays a single binding affinity for a particular antigen with which it immunoreacts.
  • Traditional full length antibody structural units typically comprise a tetramer.
  • Each tetramer is typically composed of two identical pairs of polypeptide chains, each pair having one “light” (typically having a molecular weight of about 25 kDa) and one “heavy” chain (typically having a molecular weight of about 50-70 kDa).
  • Human light chains are classified as kappa and lambda light chains.
  • the present invention is directed to the IgG class, which has several subclasses, including, but not limited to IgG1, IgG2, IgG3, and IgG4.
  • isotype as used herein is meant any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions.
  • the exemplary antibodies herein designated “CPA” are based on IgG1 heavy constant regions, as shown in DB2/ 47495312.3 22 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO Figure 4, the anti-PVRIG antibodies of the invention include those using IgG2, IgG3 and IgG4 sequences, or combinations thereof.
  • IgG2 IgG3
  • IgG4 sequences or combinations thereof.
  • different IgG isotypes have different effector functions which may or may not be desirable.
  • the CPA antibodies of the invention can also swap out the IgG1 constant domains for IgG2, IgG3 or IgG4 constant domains (depicted in Figure 4), with IgG2 and IgG4 finding particular use in a number of situations, for example for ease of manufacture or when reduced effector function is desired, the latter being desired in some situations.
  • the amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition, generally referred to in the art and herein as the “Fv domain” or “Fv region”. In the variable region, three loops are gathered for each of the V domains of the heavy chain and light chain to form an antigen- binding site.
  • CDR complementarity-determining region
  • Variable refers to the fact that certain segments of the variable region differ extensively in sequence among antibodies. Variability within the variable region is not evenly distributed. Instead, the V regions consist of relatively invariant stretches called framework regions (FRs) of 15-30 amino acids separated by shorter regions of extreme variability called “hypervariable regions”.
  • Each VH and VL is composed of three hypervariable regions (“complementary determining regions,” “CDRs”) and four FRs, arranged from amino-terminus to carboxy- terminus in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.
  • the hypervariable region generally encompasses amino acid residues from about amino acid residues 24-34 (LCDR1; “L” denotes light chain), 50-56 (LCDR2) and 89-97 (LCDR3) in the light chain variable region and around about 31-35B (HCDR1; “H” denotes heavy chain), 50-65 (HCDR2), and 95-102 (HCDR3) in the heavy chain variable region, although sometimes the numbering is shifted slightly as will be appreciated by those in the art; Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, 5 th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.
  • immunoglobulin (Ig) domain herein is meant a region of an immunoglobulin having a distinct tertiary structure.
  • the heavy chain domains including, the constant heavy (CH) domains and the hinge domains.
  • the IgG isotypes each have three CH regions.
  • “CH” domains in the context of IgG are as follows: “CH1” refers to positions 118-220 according to the EU index as in Kabat. “CH2” refers to positions 237-340 according to the EU index as in Kabat, and “CH3” refers to positions 341-447 according to the EU index as in Kabat. [0118] Accordingly, the invention provides variable heavy domains, variable light domains, heavy constant domains, light constant domains and Fc domains to be used as outlined herein.
  • variable region as used herein is meant the region of an immunoglobulin that comprises one or more Ig domains substantially encoded by any of the V ⁇ or V ⁇ , and/or VH genes that make up the kappa, lambda, and heavy chain immunoglobulin genetic loci respectively.
  • the variable heavy domain comprises vhFR1-vhCDR1-vhFR2- vhCDR2-vhFR3-vhCDR3-vhFR4
  • variable light domain comprises vlFR1-vlCDR1- vlFR2-vlCDR2-vlFR3-vlCDR3-vlFR4.
  • heavy constant region herein is meant the CH1-hinge-CH2-CH3 portion of an antibody.
  • Fc or “Fc region” or “Fc domain” as used herein is meant the polypeptide comprising the constant region of an antibody excluding the first constant region immunoglobulin domain and in some cases, part of the hinge.
  • Fc refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, the last three constant region immunoglobulin domains of IgE and IgM, and the flexible hinge N- terminal to these domains.
  • Fc may include the J chain.
  • the Fc domain comprises immunoglobulin domains C ⁇ 2 and C ⁇ 3 (C ⁇ 2 and C ⁇ 3) and the lower hinge region between C ⁇ 1 (C ⁇ 1) and C ⁇ 2 (C ⁇ 2).
  • the human IgG heavy chain Fc region is usually defined to include residues C226 or P230 to its carboxyl-terminus, wherein the numbering is according to the EU index as in DB2/ 47495312.3 24 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO Kabat.
  • amino acid modifications are made to the Fc region, for example to alter binding to one or more Fc ⁇ R receptors or to the FcRn receptor.
  • the Fc variants of the present invention are defined according to the amino acid modifications that compose them.
  • N434S or 434S is an Fc variant with the substitution serine at position 434 relative to the parent Fc polypeptide, wherein the numbering is according to the EU index.
  • M428L/N434S defines an Fc variant with the substitutions M428L and N434S relative to the parent Fc polypeptide.
  • the identity of the WT amino acid may be unspecified, in which case the aforementioned variant is referred to as 428L/434S.
  • Fab or “Fab region” as used herein is meant the polypeptide that comprises the VH, CH1, VL, and CL immunoglobulin domains. Fab may refer to this region in isolation, or this region in the context of a full length antibody, antibody fragment or Fab fusion protein.
  • Fv or “Fv fragment” or “Fv region” as used herein is meant a polypeptide that comprises the VL and VH domains of a single antibody. As will be appreciated by those in the art, these generally are made up of two chains. [0121] Throughout the present specification, either the IMTG numbering system or the Kabat numbering system is generally used when referring to a residue in the variable domain (approximately, residues 1-107 of the light chain variable region and residues 1-113 of the heavy chain variable region) (e.g, Kabat et al., supra (1991)). EU numbering as in Kabat is generally used for constant domains and/or the Fc domains.
  • the CDRs contribute to the formation of the antigen-binding, or more specifically, epitope binding site of antibodies.
  • Epitope refers to a determinant that interacts with a specific antigen binding site in the variable region of an antibody molecule known as a paratope. Epitopes are groupings of molecules such as amino acids or sugar side chains and usually have specific structural characteristics, as well as specific charge characteristics. A single antigen may have more than one epitope. DB2/ 47495312.3 25 MLB Ref.
  • the epitope may comprise amino acid residues directly involved in the binding (also called immunodominant component of the epitope) and other amino acid residues, which are not directly involved in the binding, such as amino acid residues which are effectively blocked by the specifically antigen binding peptide; in other words, the amino acid residue is within the footprint of the specifically antigen binding peptide.
  • Epitopes may be either conformational or linear. A conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain.
  • Conformational and nonconformational epitopes may be distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.
  • An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Antibodies that recognize the same epitope can be verified in a simple immunoassay showing the ability of one antibody to block the binding of another antibody to a target antigen, for example “binning”. Specific bins are described below.
  • antibody included within the definition of “antibody” is an “antigen-binding portion” of an antibody (also used interchangeably with “antigen-binding fragment”, “antibody fragment” and “antibody derivative”).
  • an antibody of the invention has a minimum functional requirement that it bind to a PVRIG antigen.
  • antigen fragments and derivatives that retain the ability to bind an antigen and yet have alternative structures, including, but not limited to, (i) the Fab fragment consisting of VL, VH, CL and CH1 domains, (ii) the Fd fragment consisting of the VH and CH1 domains, (iii) F(ab')2 fragments, a bivalent fragment comprising two linked Fab fragments (vii) single chain Fv molecules (scFv), wherein a VH domain and a VL domain are linked by a peptide linker which allows the two domains to associate to form an antigen binding site (Bird et al., 1988, Science 242:423-426, Huston et al., 1988, Proc.
  • immunoadhesion molecules include use of the streptavidin core region to make a tetrameric scFv molecule and use of a cysteine residue, a marker peptide and a C-terminal polyhistidine tag to make bivalent and biotinylated scFv molecules.
  • Antibody portions such as Fab and F(ab') 2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion, respectively, of whole antibodies.
  • antibodies, antibody portions and immunoadhesion molecules can be obtained using standard recombinant DNA techniques, as described herein. [0128]
  • the anti-PVRIG antibodies of the invention are recombinant.
  • Recombinant refers broadly with reference to a product, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified.
  • recombinant cells express genes that are not found within the native (non- recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.
  • recombinant antibody includes all antibodies that are prepared, expressed, created or isolated by recombinant means, such as (a) antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom (described further below), (b) antibodies isolated from a host cell transformed to express the human antibody, e.g., from a transfectoma, (c) antibodies isolated from a recombinant, combinatorial human antibody library, and (d) antibodies prepared, expressed, created or isolated by any other means that involve splicing of human immunoglobulin gene sequences to other DNA sequences.
  • Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences.
  • such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic DB2/ 47495312.3 27 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.
  • in vitro mutagenesis or, when an animal transgenic for human Ig sequences is used, in vivo somatic DB2/ 47495312.3 27 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO mutagenesis
  • anti-PVRIG antibodies e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3
  • the anti-PVRIG antibodies can be modified, or engineered, to alter the amino acid sequences by amino acid substitutions.
  • amino acid substitution or “substitution” herein is meant the replacement of an amino acid at a particular position in a parent polypeptide sequence with a different amino acid.
  • the substitution is to an amino acid that is not naturally occurring at the particular position, either not naturally occurring within the organism or in any organism.
  • the substitution E272Y refers to a variant polypeptide, in this case an Fc variant, in which the glutamic acid at position 272 is replaced with tyrosine.
  • a protein which has been engineered to change the nucleic acid coding sequence but not change the starting amino acid is not an “amino acid substitution”; that is, despite the creation of a new gene encoding the same protein, if the protein has the same amino acid at the particular position that it started with, it is not an amino acid substitution.
  • amino acid substitutions can be made to alter the affinity of the CDRs for the PVRIG protein (including both increasing and decreasing binding, as is more fully outlined below), as well as to alter additional functional properties of the antibodies.
  • the antibodies may be engineered to include modifications within the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen-dependent cellular cytotoxicity.
  • an antibody according to at least some embodiments of the invention may be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more functional properties of the antibody. Such embodiments are described further below.
  • the hinge region of CH1 is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased.
  • This approach is DB2/ 47495312.3 28 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO described further in U.S. Pat. No.5,677,425 by Bodmer et al.
  • the number of cysteine residues in the hinge region of CH1 is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.
  • the Fc hinge region of an antibody is mutated to decrease the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding.
  • SpA Staphylococcyl protein A
  • This approach is described in further detail in U.S. Pat. No.6,165,745 by Ward et al.
  • amino acid substitutions can be made in the Fc region, in general for altering binding to Fc ⁇ R receptors.
  • Fc gamma receptor any member of the family of proteins that bind the IgG antibody Fc region and is encoded by an Fc ⁇ R gene. In humans this family includes but is not limited to Fc ⁇ RI (CD64), including isoforms Fc ⁇ RIa, Fc ⁇ RIb, and Fc ⁇ RIc; Fc ⁇ RII (CD32), including isoforms Fc ⁇ RIIa (including allotypes H131 and R131), Fc ⁇ RIIb (including Fc ⁇ RIIb-1 and Fc ⁇ RIIb-2), and Fc ⁇ RIIc; and Fc ⁇ RIII (CD16), including isoforms Fc ⁇ RIIIa (including allotypes V158 and F158) and Fc ⁇ RIIIb (including allotypes Fc ⁇ RIIIb-NA1 and Fc ⁇ RIIIb-NA2) (Jefferis et al., 2002, Immunol Lett 82:57-65
  • An Fc ⁇ R may be from any organism, including but not limited to humans, mice, rats, rabbits, and monkeys.
  • Mouse Fc ⁇ Rs include but are not limited to Fc ⁇ RI (CD64), Fc ⁇ RII (CD32), Fc ⁇ RIII-1 (CD16), and Fc ⁇ RIII-2 (CD16-2), as well as any undiscovered mouse Fc ⁇ Rs or Fc ⁇ R isoforms or allotypes.
  • Fc ⁇ Rs include but are not limited to Fc ⁇ RI (CD64), Fc ⁇ RII (CD32), Fc ⁇ RIII-1 (CD16), and Fc ⁇ RIII-2 (CD16-2), as well as any undiscovered mouse Fc ⁇ Rs or Fc ⁇ R isoforms or allotypes.
  • ADCC antibody dependent cell-mediated cytotoxicity; the cell- mediated reaction wherein nonspecific cytotoxic cells that express Fc ⁇ Rs recognize bound antibody on a target cell and subsequently cause lysis of the target cell.
  • Fc ⁇ RIIb an inhibitory receptor
  • Amino acid substitutions that find use in the present invention include those listed in U.S. Ser. Nos.11/124,620 (particularly FIG.41) and U.S. Patent No.6,737,056, both of which are expressly incorporated herein by reference in their entirety and specifically for the variants DB2/ 47495312.3 29 MLB Ref.
  • the antibody can be altered within the C H1 or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, as described in U.S. Pat. Nos.5,869,046 and 6,121,022 by Presta et al. Additional mutations to increase serum half-life are disclosed in U.S. Patent Nos.8,883,973, 6,737,056 and 7,371,826, and include 428L, 434A, 434S, and 428L/434S.
  • the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector functions of the antibody.
  • one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320 and 322 can be replaced with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen-binding ability of the parent antibody.
  • the effector ligand to which affinity is altered can be, for example, an Fc receptor or the C1 component of complement. This approach is described in further detail in U.S. Pat. Nos.5,624,821 and 5,648,260, both by Winter et al.
  • one or more amino acids selected from amino acid residues 329, 331 and 322 can be replaced with a different amino acid residue such that the antibody has altered C1q binding and/or reduced or abolished complement dependent cytotoxicity (CDC).
  • CDC complement dependent cytotoxicity
  • one or more amino acid residues within amino acid positions 231 and 239 are altered to thereby alter the ability of the antibody to fix complement. This approach is described further in PCT Publication WO 94/29351 by Bodmer et al.
  • the Fc region is modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and/or to increase the affinity of the antibody for an Fc ⁇ receptor by modifying one or more amino acids at the following positions: 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 329, 330, 331, 333, 334, 335, 337, 338, 340, DB2/ 47495312.3 30 MLB Ref.
  • ADCC antibody dependent cellular cytotoxicity
  • the antibody can be modified to abrogate in vivo Fab arm exchange.
  • this process involves the exchange of IgG4 half-molecules (one heavy chain plus one light chain) between other IgG4 antibodies that effectively results in bispecific antibodies which are functionally monovalent. Mutations to the hinge region and constant domains of the heavy chain can abrogate this exchange (see Aalberse, RC, Schuurman J., 2002, Immunology 105:9-19).
  • the glycosylation of an antibody is modified.
  • an aglycosylated antibody can be made (i.e., the antibody lacks glycosylation).
  • Glycosylation can be altered to, for example, increase the affinity of the antibody for antigen or reduce effector function such as ADCC.
  • Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence, for example N297. For example, one or more amino acid substitutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site.
  • an antibody can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNac structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies.
  • Such carbohydrate modifications can be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery.
  • Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which to express recombinant antibodies according to at least some embodiments of the invention to thereby produce an antibody with altered glycosylation.
  • the cell lines Ms704, Ms705, DB2/ 47495312.3 31 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO and Ms709 lack the fucosyltransferase gene, FUT8 ( ⁇ (1,6) fucosyltransferase), such that antibodies expressed in the Ms704, Ms705, and Ms709 cell lines lack fucose on their carbohydrates.
  • the Ms704, Ms705, and Ms709 FUT8 cell lines are created by the targeted disruption of the FUT8 gene in CHO/DG44 cells using two replacement vectors (see U.S. Patent Publication No.20040110704 by Yamane et al. and Yamane-Ohnuki et al. (2004) Biotechnol Bioeng 87:614-22).
  • EP 1,176,195 by Hanai et al. describes a cell line with a functionally disrupted FUT8 gene, which encodes a fucosyl transferase, such that antibodies expressed in such a cell line exhibit hypofucosylation by reducing or eliminating the ⁇ 1,6 bond-related enzyme.
  • PCT Publication WO 03/035835 by Presta describes a variant CHO cell line, Lec13 cells, with reduced ability to attach fucose to Asn(297)-linked carbohydrates, also resulting in hypofucosylation of antibodies expressed in that host cell (see also Shields, R. L. et al. (2002) J. Biol. Chem.277:26733-26740).
  • PCT Publication WO 99/54342 by Umana et al.
  • glycoprotein-modifying glycosyl transferases e.g., ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnTIII)
  • GnTIII glycoprotein-modifying glycosyl transferases
  • the fucose residues of the antibody may be cleaved off using a fucosidase enzyme.
  • the fucosidase ⁇ -L-fucosidase removes fucosyl residues from antibodies (Tarentino, A. L. et al.
  • Another modification of the antibodies herein that is contemplated by the invention is pegylation or the addition of other water soluble moieties, typically polymers, e.g., in order to enhance half-life.
  • An antibody can be pegylated to, for example, increase the biological (e.g., serum) half-life of the antibody.
  • PEG polyethylene glycol
  • the antibody, or fragment thereof typically is reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment.
  • PEG polyethylene glycol
  • the pegylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer).
  • a reactive PEG molecule or an analogous reactive water-soluble polymer.
  • polyethylene glycol is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (C 1 -C 10 ) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide.
  • the antibody to be pegylated is an aglycosylated antibody.
  • an “affinity matured” antibody is one having one or more alteration(s) in one or more CDRs which results in an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess those alteration(s). In some cases, although rare, it may be desirable to decrease the affinity of an antibody to its antigen, but this is generally not preferred.
  • one or more amino acid modifications are made in one or more of the CDRs of the PVRIG antibodies of the invention. In general, only 1 or 2 or 3-amino acids are substituted in any single CDR, and generally no more than from 1, 2, 3.4, 5, 6, 7, 8 9 or 10 changes are made within a set of CDRs.
  • Affinity maturation can be done to increase the binding affinity of the antibody for the PVRIG antigen by at least about 10% to 50-100-150% or more, or from 1 to 5 fold as compared to the “parent” antibody.
  • Preferred affinity matured antibodies will have nanomolar or even picomolar affinities for the PVRIG antigen.
  • Affinity matured antibodies are produced by known procedures. See, for example, Marks et al., 1992, Biotechnology 10:779- 783 that describes affinity maturation by variable heavy chain (VH) and variable light chain (VL) domain shuffling.
  • CDRs and antibodies of the invention include amino acid modifications in one or more of the CDRs of the enumerated antibodies of the invention.
  • amino acid modifications can also independently and optionally be made in any region outside the CDRs, including framework and constant regions.
  • the present invention provides anti-PVRIG antibodies.
  • anti-PVRIG antibodies and “PVRIG antibodies” are used interchangeably.
  • the anti-PVRIG antibodies of the invention specifically bind to human PVRIG, and preferably the ECD of human PVRIG1, as depicted in Figure 3, including, e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3.
  • Specific binding for PVRIG or a PVRIG epitope can be exhibited, for example, by an antibody having a KD of at least about 10 -4 M, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10 -9 M, alternatively at least about 10- 10 M, at least about 10 -11 M, at least about 10 -12 M, or greater, where KD refers to a dissociation rate of a particular antibody-antigen interaction.
  • an antibody that specifically binds an antigen will have a KD that is 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for a control molecule relative to the PVRIG antigen or epitope.
  • the antibodies for optimal binding to PVRIG expressed on the surface of NK and T-cells, the antibodies preferably have a KD less 50 nM and most preferably less than 1 nM, with less than 0.1 nM and less than 1 pM and 0.1 pM finding use in the methods of the invention.
  • specific binding for a particular antigen or an epitope can be exhibited, for example, by an antibody having a KA or Ka for a PVRIG antigen or epitope of at least 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for the epitope relative to a control, where KA or Ka refers to an association rate of a particular antibody-antigen interaction.
  • KA or Ka refers to an association rate of a particular antibody-antigen interaction.
  • the anti-PVRIG antibodies of the invention bind to human PVRIG with a KD of 100 nM or less, 50 nM or less, 10 nM or less, or 1 nM or less (that is, higher binding affinity), or 1pM or less, wherein K D is determined by known methods, e.g. surface plasmon resonance (SPR, e.g. Biacore assays), ELISA, KINEXA, and most typically SPR at 25 ⁇ or 37 ⁇ C.
  • SPR surface plasmon resonance
  • ELISA e.g. Biacore assays
  • the invention provides antigen binding domains, including full length antibodies, which contain a number of specific, enumerated sets of 6 CDRs, as provided in Figure 3.
  • the invention provides antigen binding domains, including full length antibodies, which contain a number of specific, enumerated sets of 6 CDRs, as provided in Figure 3.
  • the invention further provides variable heavy and light domains as well as full length heavy and light chains.
  • the invention further provides variants of the above components, including variants in the CDRs, as outlined above.
  • variable heavy chains can be at least 80%, at least 90%, at least 95%, at least 98% or at least 99% identical to the “VH” sequences herein, and/or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid changes, or more, when Fc variants are used.
  • Variable light chains are provided that can be at least 80%, at least 90%, at least 95%, at least 98% or at least 99% identical to the “VL” sequences herein, and/or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid changes, or more, when Fc variants are used.
  • heavy and light chains are provided that are at least 80%, at least 90%, at least 95%, at least 98% or at least 99% identical to the “HC” and “LC” sequences herein, and/or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid changes, or more, when Fc variants are used.
  • the present invention provides antibodies, usually full length or scFv domains, that comprise the following CHA sets of CDRs, the sequences of which are shown in Figure 3: [0161] CHA.7.518.1.H4(S241P)vhCDR1, CHA.7.518.1.H4(S241P)vhCDR2, CHA.7.518.1.H4(S241P)vhCDR3, CHA.7.518.1.H4(S241P)vlCDR1, CHA.7.518.1.H4(S241P)vlCDR2, and CHA.7.518.1.H4(S241P)vlCDR3.
  • variable heavy and variable light chains can be humanized as is known in the art (with occasional variants generated in the CDRs as needed), and thus humanized variants of the VH and VL chains of Figure 3 can be generated.
  • humanized variable heavy and light domains can then be fused with human constant regions, such as the constant regions from IgG1, IgG2, IgG3 and IgG4.
  • sequences that may have the identical CDRs but changes in the variable domain (or entire heavy or light chain).
  • PVRIG antibodies include those with CDRs identical to those shown in Figure 3 or Figures 5A-5D but whose identity along the variable region can be lower, for example 90%, 95% or 98% percent identical.
  • PVRIG antibodies include those with CDRs identical to those shown in Figure 3 but whose identity along the variable region can be lower, for example 90%, 95% or 98% percent identical, and in some embodiments at least 90%, at least 95% or at least 98%.
  • the percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl.
  • the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol.48:444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available commercially), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
  • the protein sequences of the present invention can further be used as a “query sequence” to perform a search against public databases to, for example, identify related sequences.
  • Such searches can be performed using the XBLAST program (version 2.0) of Altschul, et al. (1990) J Mol. Biol.215:403-10.
  • Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402.
  • the default parameters of the respective programs e.g., XBLAST and NBLAST
  • the percentage identity for comparison between PVRIG antibodies is at least 75%, at least 80%, at least 90%, with at least about 95, 96, 97, 98 or 99% percent identity being preferred.
  • the percentage identity may be along the whole amino acid DB2/ 47495312.3 36 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO sequence, for example the entire heavy or light chain or along a portion of the chains.
  • anti-PVRIG antibodies of the invention include those that share identity along the entire variable region (for example, where the identity is 95 or 98% identical along the variable regions, and in some embodiments at least 95% or at least 98%), or along the entire constant region, or along just the Fc domain.
  • TIGIT Antibodies [0099] The present invention provides anti-TIGIT antibodies. (For convenience, “anti-TIGIT antibodies” and “TIGIT antibodies” are used interchangeably).
  • the anti- TIGIT antibodies of the invention specifically bind to human TIGIT, and preferably the ECD of human TIGIT.
  • the invention further provides antigen binding domains, including full length antibodies, which contain a number of specific, enumerated sets of 6 CDRs that bind to TIGIT, as shown for example in Figure 4.
  • Specific binding for TIGIT or a TIGIT epitope can be exhibited, for example, by an antibody having a K D of at least about 10 -4 M, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10 -9 M, alternatively at least about 10- 10 M, at least about 10 -11 M, at least about 10 -12 M, at least about 10 -13 M, at least about 10 -14 M, at least about 10 -15 M, or greater, where K D refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.
  • an antibody that specifically binds an antigen will have a K D that is 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for a control molecule relative to the TIGIT antigen or epitope.
  • the antibodies preferably have a KD less 50 nM and most preferably less than 1 nM, with less than 0.1 nM and less than 1 pM finding use in the methods of the invention
  • specific binding for a particular antigen or an epitope can be exhibited, for example, by an antibody having a ka (referring to the association rate constant) for a TIGIT antigen or epitope of at least 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for the epitope relative to a control, where ka refers to the association rate constant of a particular antibody-antigen interaction.
  • the anti-TIGIT antibodies of the invention bind to human TIGIT with a K D of 100 nM or less, 50 nM or less, 10 nM or less, or 1 nM or less (that is, higher binding affinity), or 1pM or less, wherein KD is determined by known methods, e.g. DB2/ 47495312.3 37 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO surface plasmon resonance (SPR, e.g. Biacore assays), ELISA, KINEXA, and most typically SPR at 25 ⁇ or 37 ⁇ C.
  • KD is determined by known methods, e.g. DB2/ 47495312.3 37 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO surface plasmon resonance (SPR, e.g. Biacore assays), ELISA, KINEXA, and most typically SPR at 25 ⁇ or 37 ⁇ C.
  • the invention provides antigen binding domains, including full length antibodies, which contain a number of specific, enumerated sets of 6 CDRs, as provided in Figure 3.
  • the invention provides antigen binding domains, including full length antibodies, which contain a number of specific, enumerated sets of 6 CDRs, as provided in Figure 3.
  • the invention further provides variable heavy and light domains as well as full length heavy and light chains.
  • the invention further provides variants of the above components, including variants in the CDRs, as outlined above.
  • heavy and light chains are provided that are at least 80%, at least 90%, at least 95%, at least 98% or at least 99% identical to the “HC” and “LC” sequences herein, and/or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid changes, or more, when Fc variants are used.
  • the present invention provides antibodies, usually full length or scFv domains, that comprise the following CHA sets of CDRs, the sequences of which are shown in Figure 4: CPA.9.083.H4(S241P)vhCDR1, CPA.9.083.H4(S241P)vhCDR2, CPA.9.083.H4(S241P)vhCDR3, CPA.9.083.H4(S241P) vlCDR1, CPA.9.083.H4(S241P) vlCDR2, and CPA.9.083.H4(S241P)vlCDR3; or CPA.9.086.H4(S241P)vhCDR1, CPA.9.086.H4(S241P)vhCDR2, CPA.9.086.H4(S241P)vhCDR3, CPA.9.086.H4(S241P) vlCDR1, CPA.9.086.
  • TIGIT antibodies include those with CDRs identical to those shown in Figure 4 but whose identity along the variable region can be lower, for example 95 or 98% percent identical.
  • TIGIT antibodies include those with CDRs identical to those shown in Figure 5 but whose identity along the variable region can be lower, for example 95 or 98% percent identical, and in some embodiments at least 95% or at least 98%.
  • the percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17 (1988)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol.
  • the protein sequences of the present invention can further be used as a “query sequence” to perform a search against public databases to, for example, identify related sequences. Such searches can be performed using the XBLAST program (version 2.0) of Altschul, et al. (1990) J Mol. Biol.215:403-10.
  • Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402.
  • the default parameters of the respective programs e.g., XBLAST and NBLAST
  • the percentage identity for comparison between TIGIT antibodies is at least 75%, at least 80%, at least 90%, with at least about 95, 96, 97, 98 or 99% percent identity being preferred.
  • the antibodies of the invention are immunomodulatory, in that rather than directly attack cancerous cells, the anti-TIGIT and anti-PVRIG antibodies of the invention stimulate the immune system, generally by inhibiting the action of TIGIT and PVRIG, respectively.
  • cancer immunotherapy is aimed to stimulate the patient’s or subject’s own immune system to eliminate cancer cells, providing long-lived tumor destruction.
  • the anti-TIGIT, anti-PVRIG, and pembrolizumab combination of the invention are useful in treating cancer. Due to the nature of an immuno-oncology mechanism of action, TIGIT and or PVRIG do not necessarily need to be overexpressed on or DB2/ 47495312.3 40 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO correlated with a particular cancer type; that is, the goal is to have the anti-TIGIT antibodies de-suppress T cell and NK cell activation, such that the immune system will go after the cancers. VII.
  • compositions comprising a carrier suitable for the desired delivery method.
  • Suitable carriers include any material that when combined with the therapeutic composition retains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient’s or subject’s immune system.
  • Examples include, but are not limited to, any of a number of standard pharmaceutical carriers such as sterile phosphate buffered saline solutions, bacteriostatic water, and the like (see, generally, Remington's Pharmaceutical Sciences 16 th Edition, A. Osal., Ed., 1980).
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, acetate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl orbenzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, hist
  • the pharmaceutical composition that comprises anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3) of the invention may be in a water-soluble form, such as being present as pharmaceutically acceptable salts, which is meant to include both acid and base addition salts.
  • “Pharmaceutically acceptable acid addition salt” refers to those salts that retain the biological effectiveness of the free bases and that are not biologically or otherwise undesirable, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like
  • organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid,
  • “Pharmaceutically acceptable base addition salts” include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Particularly preferred are the ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non- toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. The formulations to be used for in vivo administration are preferrably sterile.
  • the term “activity” refers to a functional activity or activities of anti- PVRIG antibodies and/or antigen binding portions thereof. Functional activities include, but are not limited to, biological activity and binding affinity.
  • the term “stability” is used in a structural context, e.g., relating to the structural integrity of an anti-PVRIG antibody and/or antigen binding portion thereof, or in a functional context, e.g., relating to a an anti-PVRIG antibody and/or antigen binding portion thereof 's ability to retain its function and/or activity over time (e.g., including anti-PVRIG antibody and/or antigen binding portion thereof stability or anti-PVRIG antibody and/or antigen binding portion thereof formulation stability, wherein the anti-PVRIG antibody includes those with CDRs identical to those shown in Figure 3).
  • the anti-PVRIG antibody and/or antigen binding portion thereof under discussion may be contained within a formulation in accordance with the methods and compositions described herein, and the stability of that protein refers to its stability in that formulation.
  • the stability of an anti-PVRIG antibody and/or antigen binding portion thereof composition is determined by measuring the binding activity of the composition, including for example, using the assays described in the application and figures provided herewith, as DB2/ 47495312.3 42 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO well as any other applicable assays known in the art.
  • the stability of an anti-PVRIG antibody and/or antigen binding portion thereof composition is formulated with sugar, sugar alcohol, and/or non-ionic surfactant, as described herein, is compared to an anti- PVRIG antibody and/or antigen binding portion thereof composition formulated without the at least one amino acid, salt, and/or non-ionic surfactant and/or with a different combination of components.
  • the formulation does not comprise a sugar and/or sugar alcohol.
  • a “storage stable” aqueous an anti-PVRIG antibody and/or antigen binding portion thereof composition refers to a an anti-PVRIG antibody and/or antigen binding portion thereof comprising solution that has been formulated to increase the stability of the protein in solution, for example by at least 10%, over a given storage time.
  • an anti-PVRIG antibody and/or antigen binding portion thereof can be made “storage stable” by the addition of at least one amino acid, salt, or non- ionic surfactant as a stabilizing agent.
  • the stability of the anti-PVRIG antibody and/or antigen binding portion thereof in any given formulation can be measured, for example, by monitoring the formation of aggregates, loss of bulk binding activity, or formation of degradation products, over a period of time.
  • the absolute stability of a formulation, and the stabilizing effects of the sugar, sugar alcohol, or non-ionic surfactant, will vary dependent upon the particular composition being stabilized.
  • the stability of an anti-PVRIG antibody and/or antigen binding portion thereof composition is determined by measuring the anti-PVRIG antibody and/or antigen binding portion thereof binding activity of the composition. For example, by using an ELISA or other binding activity assay.
  • the stability of an anti-PVRIG antibody and/or antigen binding portion thereof composition formulated with sugar, sugar alcohol, and/or non-ionic surfactant, as described herein, is compared to an anti-PVRIG antibody and/or antigen binding portion thereof composition formulated without the a least one amino acid, salt, and/or non-ionic surfactant and/or with a different combination of components.
  • the formulation does not comprise a sugar and/or sugar alcohol.
  • the predetermined temperature refers to frozen (e.g., -80°C, -25°C, 0°C), refrigerated (e.g., 0° to 10°C), or room temperature (e.g., 18°C to 32° C) storage.
  • DB2/ 47495312.3 43 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO the term “time of stability” refers to the length of time a formulation is considered stable.
  • the time of stability for a formulation may refer to the length of time for which the level of protein aggregation and/or degradation in the formulation remains below a certain threshold (e.g., 1 %, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, etc.), and/or the length of time a formulation maintains biological activity above a certain threshold (e.g., 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, etc.) of the amount of activity (including, for example, binding activity) present in the formulation at the start of the storage period.
  • a certain threshold e.g., 1 %, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%,
  • a storage stable aqueous composition of a an anti-PVRIG antibody and/or antigen binding portion thereof formulated with a sugar, sugar alcohol, and/or non-ionic surfactant will have a longer time of stability than a composition of the same an anti-PVRIG antibody and/or antigen binding portion thereof formulated without the at least one amino acid, salt, and/or non-ionic surfactant.
  • a storage stable aqueous composition of an anti-PVRIG antibody and/or antigen binding portion thereof will have a time of stability that is, for example, at least 10% greater than the time of stability for an anti-PVRIG antibody and/or antigen binding portion thereof composition formulated in the absence of the at least one amino acid, salt, and/or non-ionic surfactant, or at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190% greater, or at least 2 times greater, or at least 2.5 times, 3.0 times, 3.5 times, 4.0 times, 4.5 times, 5.0 times, 5.5 times, 6.0 times, 6.5 times, 7.0 times, 7.5 times, 8.0 times, 8.5 times, 9.0 times, 9.5 times, 10 times, or more times greater than the time of stability for the anti
  • BDS refers to “Bulk Drug Substance.”
  • A. Stabilized Liquid Formulations [0176] In some embodiments, the present disclosure provides stabilized aqueous formulations of an anti-PVRIG antibody and/or antigen binding portion thereof (e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3). The following embodiments are based in part on the discovery that inclusion of at least one amino acid, salt, and/or non-ionic surfactant stabilizes the liquid anti-PVRIG antibody and/or antigen binding portion thereof compositions, as compared to compositions lacking the at DB2/ 47495312.3 44 MLB Ref.
  • an anti-PVRIG antibody and/or antigen binding portion thereof formulated according to the embodiments provided herein may contain, in addition to the components explicitly disclosed, counter ions contributed by the inclusion of solution components or pH modifying agents, for example, sodium or potassium contributed from an acetate salt, sodium hydroxide, or potassium hydroxide or chloride contributed by calcium chloride or hydrochloric acid.
  • a storage stable an anti-PVRIG antibody and/or antigen binding portion thereof composition consisting of or consisting essentially of a given formulation may further comprise one or more counter ion, as necessitated by the formulation process at a particular pH.
  • a storage stable anti-PVRIG antibody and/or antigen binding portion provided herein will be stabilized at cryopreservation temperature (i.e., -20°C or below) for a period of time.
  • cryopreservation temperature i.e., -20°C or below
  • a stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof will be stable when stored at cryopreservation temperature for at least 4 days.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable at cryopreservation temperature for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 21, 28, or more days. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, or more. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 month.
  • the composition will be stable for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months.
  • the anti- PVRIG antibody and/or antigen binding portion composition will be stable for an extended period of time when stored at a temperature of about -20°C or below.
  • a storage stable anti-PVRIG antibody and/or antigen binding portion provided herein will be stabilized at refrigerated temperature (i.e., between 2°C and 10°C) for a period of time.
  • a stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof will be stable when stored at refrigerated temperature for at least 4 days.
  • the DB2/ 47495312.3 45 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO anti-PVRIG antibody and/or antigen binding portion composition will be stable at refrigerated temperature for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 21, 28, or more days.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, or more.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 month. In some embodiments, the composition will be stable for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion composition will be stable for an extended period of time when stored at a temperature between 2°C and 8°C.
  • a stable liquid pharmaceutical formulations comprising an anti- PVRIG antibody or antigen binding fragment thereof provided herein will be stabilized at room temperature (i.e., between 18°C and 32°C) for a period of time.
  • room temperature i.e., between 18°C and 32°C
  • a stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof will be stable when stored at room temperature for at least 4 days.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable at room temperature for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 21, 28, or more days.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, or more. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 month. In yet other embodiments, the composition will be stable for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months.
  • room temperature refers to between 20°C and 30°C, between 21°C and 29°C, between 22°C and 28°C, between 23°C and 27°C, between 24°C and 26°C, or about 25°C.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for an extended period of time when stored at a temperature between 20°C and 25°C. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion composition will be stable for an extended period of time when stored at a temperature of about 25°C.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion composition will be stable for at least 1 month. In yet other embodiments, the anti- PVRIG antibody and/or antigen binding portion composition will be stable for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months.
  • the anti-PVRIG antibody and/or antigen binding portion composition will be stable for an extended period of time when stored at a temperature between 35°C and 40°C.
  • antibody binding activity is measure using any assay known in the art.
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., at least one amino acid, salt, and/or non-ionic surfactant) when the anti-PVRIG antibody and/or antigen binding portion composition contains at least 10% more antibody binding activity after storage for a period of time, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • a stabilizing agent e.g., at least one amino acid, salt, and/or non-ionic surfactant
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., at least one amino acid, salt, and/or non-ionic surfactant) when the composition contains at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a greater percentage more anti-PVRIG antibody and/or antigen binding portion activity after storage for a period of time, as compared to an DB2/ 47495312.3 47 MLB Ref.
  • a stabilizing agent e.g., at least one amino acid, salt, and/or non-ionic surfactant
  • a stored anti-PVRIG antibody and/or antigen binding portion composition is considered stable as long as the percentage of anti-PVRIG antibody and/or antigen binding portion present in an aggregated state remains no more than 50%.
  • a stored anti-PVRIG antibody and/or antigen binding portion thereof composition is considered stable as long as the percentage of the anti-PVRIG antibody and/or antigen binding portion thereof present in an aggregated state remains no more than 45%, 40%, 35%, 30%, 25%, 24%, 23%, 22%, 21 %, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, or less.
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (anti- PVRIG antibody and/or antigen binding portion composition, at least one amino acid, salt, and/or non-ionic surfactant) when the composition contains at least 10% less anti-PVRIG antibody and/or antigen binding portion present in an aggregated state after storage for a period of time, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • a stabilizing agent antioxidant, at least one amino acid, salt, and/or non-ionic surfactant
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., at least one amino acid, salt, and/or non-ionic surfactant) when the composition contains at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a greater percentage less anti-PVRIG antibody and/or antigen binding portion present in an aggregated state after storage for a period of time, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent [0186]
  • the mechanical stress is agitation (e.g., shaking).
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., at least one amino acid, salt, or non-ionic surfactant) when the anti-PVRIG antibody and/or antigen binding portion composition contains at least 10% more binding activity after being subjected to mechanical stress, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • a stabilizing agent e.g., at least one amino acid, salt, or non-ionic surfactant
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., a sugar, sugar alcohol, or non-ionic surfactant) when the composition contains at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a greater percentage more furin activity after being subjected to mechanical stress, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • the mechanical stress is agitation (e.g., shaking).
  • a stored anti-PVRIG antibody and/or antigen binding portion composition is considered stable as long as the percentage of anti-PVRIG antibody and/or antigen binding portion present in an aggregated state remains no more than 50% after being subjected to mechanical stress.
  • a stored anti-PVRIG antibody and/or antigen binding portion composition is considered stable as long as the percentage of anti- PVRIG antibody and/or antigen binding portion present in an aggregated state remains no more than 45%, 40%, 35%, 30%, 25%, 24%, 23%, 22%, 21 %, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less after being subjected to mechanical stress.
  • the mechanical stress is agitation (e.g., shaking).
  • an anti-PVRIG antibody and/or antigen binding portion composition is considered to have been stabilized by the addition of a stabilizing agent (e.g., at least one amino acid, salt, or non-ionic surfactant) when the composition contains at least 10% less anti-PVRIG antibody and/or antigen binding portion present in an aggregated state after being subjected to mechanical stress, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • a stabilizing agent e.g., at least one amino acid, salt, or non-ionic surfactant
  • No.114386-5030-WO Compugen Ref.230-WO stabilizing agent e.g., at least one amino acid, salt, or non-ionic surfactant
  • the composition contains at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a greater percentage less anti-PVRIG antibody and/or antigen binding portion present in an aggregated state after being subjected to mechanical stress, as compared to an anti-PVRIG antibody and/or antigen binding portion composition not containing the stabilizing agent or containing a lower amount of the stabilizing agent.
  • the mechanical stress is agitation (e.g., shaking).
  • the highly stabilized formulations of the invention have a shelf life of at least 6 months. As will be appreciated, this shelf life may be at frozen temperatures (i.e., -80°C, -25°C, 0°C), refrigerated (0°C to 10°C), or room temperature (20°C to 32°C) in liquid or lyophilized form. I n further aspects, the highly stabilized formulations of the invention have a shelf life of at least 12, 18, 24, 30, 36, 42, 48, 54, or 60 months. [0191] In some embodiments, shelf life is determined by a percent activity remaining after storage at any of the above temperatures for any of the above periods of time.
  • shelf life means that the formulation retains at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% of furin activity as measured by any of the assays described herein or known in the art as compared to activity prior to storage for any of the above amounts of time at any of the above temperatures.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody comprising: (a) an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises an antibody with CDRs identical to those shown in Figure 3; (b) 25 mM histidine; (c) 60 mM NaCl; (d) 100 mM L-Arginine, and (e) 0.01% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0.
  • the anti-PVRIG antibody is at a concentration of from 10 mg/mL to 40 mg/mL, 15 mg/mL to 40 mg/mL, 15 mg/mL to 30 mg/mL, 10 mg/mL to 25 mg/mL, or 15 mg/mL to 25 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 10 mg/mL to 40 mg/mL. In some embodiments, the anti-PVRIG DB2/ 47495312.3 50 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO antibody is at a concentration of from 15 mg/mL to 40 mg/mL.
  • the anti-PVRIG antibody is at a concentration of from 15 mg/mL to 30 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 10 mg/mL to 25 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 15 mg/mL to 25 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 10 mg/mL to 25 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 15 mg/mL to 25 mg/mL. In some embodiments, the anti-PVRIG antibody is at a concentration of from 20 mg/mL to 25 mg/mL.
  • the anti-PVRIG antibody is at a concentration of about 20 mg/mL.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody comprising: (a) an anti-PVRIG antibody, wherein the anti-PVRIG antibody comprises an antibody with CDRs identical to those shown in Figure 3; (b) from 10 mM to 100 mM histidine; (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine, and (e) from 0.005% to 0.1% w/v polysorbate 80 wherein the composition has a pH from 5.5 to 7.0. B.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3) comprising at least one amino acid.
  • the at least one amino acid is histidine.
  • the at least one amino acid is arginine.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof comprising at least two amino acids.
  • the at least two amino acids are histidine and arginine.
  • the pharmaceutical formulation comprises from 10 mM to 80 mM histidine, from 15 mM to 70 mM histidine, from 20 mM to 60 mM histidine, from 20 mM to 50 mM histidine, or from 20 mM to 30 mM histidine.
  • the DB2/ 47495312.3 51 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO pharmaceutical formulation comprises from 10 mM to 80 mM histidine.
  • the pharmaceutical formulation comprises from 15 mM to 70 mM histidine.
  • the pharmaceutical formulation comprises from 20 mM to 60 mM histidine.
  • the pharmaceutical formulation comprises from 20 mM to 50 mM histidine. In some embodiments, the pharmaceutical formulation comprises from 20 mM to 30 mM histidine. In some embodiments, the pharmaceutical formulation comprises about 25 mM histidine. [0197] In some embodiments, the pharmaceutical formulation comprises from 10 mM to 80 mM histidine. In some embodiments, the pharmaceutical formulation comprises from 15 mM to 70 mM histidine. In some embodiments, the pharmaceutical formulation comprises from 20 mM to 60 mM histidine. In some embodiments, the pharmaceutical formulation comprises from 20 mM to 50 mM histidine. In some embodiments, the pharmaceutical formulation comprises from 20 mM to 30 mM histidine.
  • the pharmaceutical formulation comprises about 25 mM histidine. [0198] In some embodiments, the pharmaceutical formulation comprises from 20 mM to 140 mM L-arginine, from 30 mM to 140 mM L-arginine, from 40 mM to 130 mM L-arginine, from 50 mM to 120 mM L-arginine, from 60 mM to 110 mM L-arginine, from 70 mM to 110 mM L-arginine, from 80 mM to 110 mM L-arginine, or from 90 mM to 110 mM L-arginine.
  • the pharmaceutical formulation comprises from 30 mM to 140 mM L-arginine. In some embodiments, the pharmaceutical formulation comprises from 40 mM to 130 mM L-arginine. In some embodiments, the pharmaceutical formulation comprises from 50 mM to 120 mM L-arginine. In some embodiments, the pharmaceutical formulation comprises from 60 mM to 110 mM L-arginine. In some embodiments, the pharmaceutical formulation comprises from 70 mM to 110 mM L-arginine. In some embodiments, the pharmaceutical formulation comprises from 80 mM to 110 mM L-arginine. DB2/ 47495312.3 52 MLB Ref.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3) comprising no sugar.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3) comprising no sugar alcohol.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof comprising a sugar and/or sugar alcohol.
  • the sugar is trehalose or sucrose.
  • the sugar is trehalose. In some embodiments, the sugar is sucrose. In some embodiments, the sugar is only one of trehalose or sucrose but not both. [0202] In some embodiments, the sugar is in an amount of from about 0.5% to 10%, 1 % to 9.5%, 1.5% to 9%, 2.0% to 8.5%, 2.5% to 8%, 3.0% to 7.5%, 3.5% to 7%, 4.0% to 6.5%, 4.5% to 6%, and/or 4.5% to 5.5%. In some embodiments, the sugar is in an amount of from about 0.5% to 10%. In some embodiments, the sugar is in an amount of from about 1 % to 9.5%. In some embodiments, the sugar is in an amount of from about 1.5% to 9%.
  • the sugar is in an amount of from about 2.0% to 8.5%. In some embodiments, the sugar is in an amount of from about 2.5% to 8%. In some embodiments, the sugar is in an amount of from about 3.0% to 7.5%. In some embodiments, the sugar is in an amount of from about 3.5% to 7%. In some embodiments, the sugar is in an amount of from about 4.0% to 6.5%. In some embodiments, the sugar is in an amount of from about 4.5% to 6%. In some embodiments, the sugar is in an amount of from about 4.5% to 5.5%. In some embodiments, the sugar is in an amount of about 5% DB2/ 47495312.3 53 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO D.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3) comprising a non-ionic surfactant.
  • an anti-PVRIG antibody or antigen binding fragment thereof e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3
  • a non-ionic surfactant e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3
  • the storage stable compositions of an anti-PVRIG antibody or antigen binding fragment comprise a non-ionic surfactant selected from a non-ionic water soluble monoglyceride, a non-ionic water soluble diglyceride, a non- ionic water soluble triglyceride, a non-ionic water soluble monofatty acid esters of polyethyelene glycol, a non-ionic water soluble difatty acid esters of polyethyelene glycol, a non-ionic water soluble sorbitan fatty acid ester, a non-ionic polyglycolyzed glyceride, a non- ionic water soluble triblock copolymer, and a combination thereof.
  • a non-ionic surfactant selected from a non-ionic water soluble monoglyceride, a non-ionic water soluble diglyceride, a non- ionic water soluble triglyceride, a non-ionic water soluble mono
  • the non-ionic surfactant is polysorbate 80 (polyoxyethylene (20) sorbitan monooleate).
  • the stable liquid pharmaceutical formulation comprises from 0.006% to 0.1% w/v polysorbate 80, from 0.007% to 0.09% w/v polysorbate 80, from 0.008% to 0.08% w/v polysorbate 80, from 0.009% to 0.09% w/v polysorbate 80, from 0.01% to 0.08% w/v polysorbate 80, from 0.01% to 0.07% w/v polysorbate 80, from 0.01% to 0.07% w/v polysorbate 80, or from 0.01% to 0.06% w/v polysorbate 80, or from 0.009% to 0.05% w/v polysorbate 80.
  • the stable liquid pharmaceutical formulation comprises from 0.006% to 0.1% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.007% to 0.09% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.008% to 0.08% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.009% to 0.09% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.01% to 0.08% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.01% to 0.07% w/v polysorbate 80.
  • the stable liquid pharmaceutical formulation comprises from 0.01% to 0.07% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.01% to 0.06% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises from 0.009% to 0.05% w/v polysorbate 80. In some embodiments, the stable liquid pharmaceutical formulation comprises about 0.01% polysorbate 80. DB2/ 47495312.3 54 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO E.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3) comprising a salt, for example, a pharmaceutically acceptable salt.
  • the stable liquid pharmaceutical formulation comprising an anti-PVRIG antibody or antigen binding fragment thereof provided herein include a pharmaceutically acceptable salt at a concentration tolerated by an anti-PVRIG antibody or antigen binding fragment thereof during storage.
  • the pharmaceutically acceptable salt is a chloride salt.
  • the pharmaceutically acceptable salt is a monovalent chloride salt.
  • the pharmaceutically acceptable salt is sodium chloride, potassium chloride, or a combination thereof.
  • the stable liquid pharmaceutical formulation comprises from 30 mM to 100 mM NaCl, from 30 mM to 90 mM NaCl, from 40 mM to 80 mM NaCl, from 30 mM to 70 mM histidine, or from 45 mM to 70 mM NaCl.
  • the stable liquid pharmaceutical formulation comprises from 30 mM to 100 mM NaCl.
  • the stable liquid pharmaceutical formulation comprises from 30 mM to 90 mM NaCl.
  • the stable liquid pharmaceutical formulation comprises from 40 mM to 80 mM NaCl.
  • the stable liquid pharmaceutical formulation comprises from 30 mM to 70 mM histidine. In some embodiments, the stable liquid pharmaceutical formulation comprises or from 45 mM to 70 mM NaCl. In some embodiments, pharmaceutical formulation comprises about 60 mM NaCl.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti- PVRIG antibodies including those with CDRs identical to those shown in Figure 3) that is buffered at a physiologically acceptable pH. In some embodiments, the physiologically acceptable pH is from about 6.0 to about 7.0.
  • stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof has a pH of from 6 to 7.0.
  • stable liquid pharmaceutical formulation of an anti-PVRIG antibody or antigen binding fragment thereof has a pH of 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0.
  • the pH is from 6.1 to 6.9.
  • the pH is from 6.2 to 6.9.
  • the pH is from 6.3 to 6.8.
  • the pH is from 6.3 to 6.7.
  • the pH is from 6.4 to 6.8. In some embodiments, the pH is from 6.5 to 6.8. In some embodiments, the pH is from 6.6 to 6.8. In some embodiments, the pH is 6.3, 6.4, 6.5, 6.6, or 6.7. In some embodiments, the pH is 6.5 +/- 0.2.
  • the method includes adding a dilution buffer, to form a diluted stable liquid pharmaceutical formulation comprising an anti-PVRIG antibody or antigen binding fragment thereof (e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3).
  • the dilution buffer is added at a ratio of from 1:1 (dilution buffer:formulation) to 1000:1 (dilution buffer:formulation). In another embodiment, the dilution buffer is added at a ratio of from 1:1 dilution buffer:formulation) to 500:1 (dilution buffer:formulation). In another embodiment, the dilution buffer is added at a ratio of from 1:1 (dilution buffer:formulation) to 250:1 (dilution buffer:formulation). In another embodiment, the dilution buffer is added at a ratio of from 1:1 (dilution buffer:formulation) to 200:1 (dilution buffer:formulation).
  • the dilution buffer is added at a ratio of from 1:1 (dilution buffer:formulation) to 100:1 (dilution buffer:formulation). In another embodiment, the dilution buffer is added at a ratio of from 1:1 (dilution buffer:formulation) to 50:1 (dilution buffer:formulation).
  • the stable liquid pharmaceutical formulation comprising an anti-PVRIG antibody or antigen binding fragment thereof is diluted from 1-fold to 1000-fold, from 1-fold to 500-fold, from 1-fold to 250-fold, from 1-fold to 200-fold, from 1-fold to 100- fold, from 1-fold to 50-fold, from 1-fold to 10-fold, from 10-fold to 1000-fold, from 10-fold to 500-fold, from 10-fold to 250-fold, from 10-fold to 200-fold, from 10-fold to 100-fold, from 10-fold to 50-fold, from 50-fold to 1000-fold, from 50-fold to 500-fold, from 50-fold to 250-fold, from 50-fold to 200-fold, from 50-fold to 100-fold, from 100-fold to 1000-fold, from 100-fold to 500-fold, from 100-fold to 250-fold, from 100-fold to 200-fold, from 200- fold to 1,000-fold, from 200-fold to 500-fold, or from 200-fold to 250-fold.
  • the stable liquid pharmaceutical formulations comprising an anti- PVRIG antibody or antigen binding fragment thereof (e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3) show improved stability as compared to control formulations.
  • improved stability includes retention of a higher percentage of binding activity and/or no reduction in binding activity as compared to control formulations in various stability assays.
  • Such assays can be used to determine if a formulation is a highly stabilized formulation.
  • the highly stabilized formulation has at least 5%, 10%, 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or greater activity than a control formulation when assessed by any of the stability assays discussed herein or known in the art.
  • the liquid pharmaceutical formulations comprising an anti- PVRIG antibody or antigen binding fragment thereof are tested under stressor conditions, such as storage at high temperature, agitation, freeze/thaw cycles, or some combination thereof. After such stressors, the formulations are assayed using any of the methods described herein or known in the art to determine the stability under these conditions.
  • an A280 by SoloVPE assay is used to examine the appearance of the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof.
  • the SoloVPE assay can be employed to examine concentrations for the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof.
  • A280 Amino acids containing aromatic side chains exhibit strong UV-light absorption at the wavelength of 280nm. Once an absorptivity coefficient has been established for a given protein, the protein’s concentration in solution can be calculated from its absorbance.
  • the method is designed to determine the protein concentration by measuring its absorbance at 280nm using the SoloVPE instrument without dilution (https://www.ctechnologiesinc.com/products/solovpe) [0218]
  • Appearance Sample appearance determination is assessed by holding the sample within a controlled light source and observe the appearance of the material. Gently agitate the solution and determine if the appearance changes when viewed against a black and white DB2/ 47495312.3 57 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO background.
  • Use adjectives such as “clear”, “turbid”, or “slightly turbid” to assess clarity. Be specific with regards to the color of the material.
  • a binding assay can be performed to examine the activity of the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof.
  • LabChip Analysis In some embodiments, a LabChip analysis is employed to examine purity, including for example, IgG purity as well as HC + LC percentages for the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof.
  • the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit IgG purity percentages greater than 94%, greater than 95%, greater than 96%, greater than 97%, or greater than 98%. In some embodiments, the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit IgG purity percentages were from about 95% to 98%. In some embodiments, the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit IgG purity percentages from about 96% to 97%. In some embodiments, the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit HC+LC percentages from about 96% to 100%.
  • the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit HC+LC percentages from about 97% to 100%. In some embodiments, the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof exhibit HC+LC percentages from about 98% to 100%.
  • cIEF Analysis [0221] In some embodiments, a capillary isoelectric focusing (cIEF) can be employed to analyze the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof for the presence of additional species, including for example, minor acidic species. DB2/ 47495312.3 58 MLB Ref.
  • Antibodies can form sub-visible particles in response to stressed conditions, such as heat, freeze/thaw cycles, and agitation.
  • a microflow imaging (MFI) analysis can be employed to analyze the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof for the formation of particles in response to stressed conditions.
  • the stable liquid pharmaceutical formulations of the anti-PVRIG antibody or antigen binding fragment thereof provide for a formulation capable of stabilizing the anti-PVRIG antibody or antigen binding fragment thereof against these stressed conditions and protecting against the formation of particles.
  • MFI can be used to evaluate particle counts at different size ranges ( ⁇ 2 ⁇ m, ⁇ 5 ⁇ m, ⁇ 10 ⁇ m, and ⁇ 25 ⁇ m) in different formulations under stressed conditions. Typically, MFI data can be evaluated to choose an appropriate formulation based on generation of the lowest amount of particles/mL for all sizes of particles across all time points, conditions, and formulations.
  • SEC Analysis [0223] In some embodiments, size exclusion chromatography (SEC) can be employed to analyze the stable liquid pharmaceutical formulations comprising an anti-PVRIG antibody or antigen binding fragment thereof. The SEC data showed HMW throughout all time points and conditions; however, it remained stable at about 1%. LMW was present in accelerated conditions and 2–8 °C 8 week time point.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or DB2/ 47495312.3 59 MLB Ref.
  • the present invention provides a stable liquid pharmaceutical formulation of an anti-PVRIG antibody comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); (b) from 10 mM to 100 mM histidine; (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); DB2/ 47495312.3 61 MLB Ref.
  • No.114386-5030-WO Compugen Ref.230-WO (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2.
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and DB2/ 47495312.3 63 MLB Ref.
  • the present invention provides a stable liquid pharmaceutical formulation comprising: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4(S241P) (SEQ ID NO:18); and ii) a light chain comprising the light chain from CHA.7.538.1.H4(S241P) (SEQ ID NO:23); (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/-
  • anti-PVRIG antibodies of the present invention e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3
  • a sterile aqueous solution may be used in a variety of ways.
  • protein therapeutics are often delivered by IV infusion.
  • the antibodies of the present invention may also be delivered using such methods.
  • administration may venious be by intravenous infusion with 0.9% sodium chloride as an infusion vehicle.
  • the dosing amounts and frequencies of administration are, in some embodiments, selected to be therapeutically or prophylactically effective.
  • adjustments for protein degradation, systemic versus localized delivery, and rate of new protease synthesis, as well as the age, body weight, general health, sex, diet, time of administration, drug interaction and the severity of the condition may be necessary, and will be ascertainable with routine experimentation by those skilled in the art.
  • a therapeutically effective dose of the Fc variant of the present invention may be administered.
  • therapeutically effective dose herein is meant a dose that produces the effects for which it is administered.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations of the present invention can be formulated for administration, including as a unit dosage formulation.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.01 mg/kg of the anti- PVRIG antibody and/or antigen binding portion thereof.
  • the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.02 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.03 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.04 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at DB2/ 47495312.3 65 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO a dosage of 0.05 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.06 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.07 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.08 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.09 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.1 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.2 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.3 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.5 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 0.8 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 1 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 2 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 3 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 4 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 5 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- DB2/ 47495312.3 66 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 6 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 7 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 8 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 9 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 10 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 15 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof. In some embodiments, the anti- PVRIG antibody and/or antigen binding portion thereof formulations are administered at a dosage of 20 mg/kg of the anti-PVRIG antibody and/or antigen binding portion thereof.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations is administered at a dosage of about 0.01 mg/kg to about 20 mg/kg of the anti-PVRIG antibody. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations is administered at a dosage of about 0.01 mg/kg to about 10 mg/kg of the anti-PVRIG antibody. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations is administered at a dosage of about 20mg/kg. In some embodiments, the anti-PVRIG antibody and/or antigen binding portion thereof formulations is administered at a dosage of about 20mg/kg each 4 weeks.
  • the anti-PVRIG antibody and/or antigen binding portion thereof formulations is administered at a dosage of about 20mg/kg IV each 4 weeks. In some embodiments, formulation is administered at a dosage of about 0.1 mg/kg to about 10 mg/kg of the anti- PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 1 mg/kg to about 10 mg/kg of the anti-PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 2 mg/kg to about 10 mg/kg of the anti-PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 3 mg/kg to about 10 mg/kg of the anti-PVRIG antibody.
  • formulation is administered at a dosage of about 4 mg/kg to about 10 mg/kg of the anti-PVRIG antibody.
  • formulation is administered at a dosage of about 5 mg/kg to about 10 mg/kg of the anti-PVRIG antibody.
  • formulation is administered at a dosage of about 5 mg/kg to about 10 mg/kg of the anti-PVRIG antibody.
  • formulation is administered at a dosage of about 7 mg/kg to about 10 mg/kg of the anti- PVRIG antibody.
  • formulation is administered at a dosage of about 8 mg/kg to about 10 mg/kg of the anti-PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 9 mg/kg to about 10 mg/kg of the anti-PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 15 mg/kg of the anti- PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 20 mg/kg of the anti-PVRIG antibody. In some embodiments, formulation is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg of the anti-PVRIG antibody.
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of
  • No.114386-5030-WO Compugen Ref.230-WO (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0.
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.518.1.H4(S241
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- DB2/ 47495312.3 69 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9);
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or DB2/ 47495312.3 70 MLB Ref.
  • an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain
  • a heavy chain variable domain is from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2.
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from
  • No.114386-5030-WO Compugen Ref.230-WO a) a VL-CL, wherein the VL from CHA.7.538.1.H4(S241P) (SEQ ID NO:19) and wherein the CL region is from human kappa 2 light chain; (b) from 10 mM to 100 mM histidine; (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0.
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from
  • No.114386-5030-WO Compugen Ref.230-WO (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2.
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4(S241P) (SEQ ID NO:18
  • the present invention provides for administration of a stable liquid pharmaceutical formulation of an anti-PVRIG antibody in combination with an anti- TIGIT antibody and pembrolizumab, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, DB2/ 47495312.3 73 MLB Ref.
  • the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2.
  • the formulation is administered with pembrolizumab. In some embodiments of the stable liquid pharmaceutical formulation, the formulation is administered with pembrolizumab. In some embodiments of the stable liquid pharmaceutical formulation, the formulation is administered with pembrolizumab, as shown in Figure 5.
  • the anti-PVRIG antibodies e.g., anti-PVRIG antibodies including those with CDRs identical to those shown in Figure 3 find use in treating patients or subjects, such as human subjects, generally with a condition associated with PVRIG.
  • treatment refers to both therapeutic treatment and prophylactic or preventative measures, which in this example relates to treatment of cancer; however, also as described below, uses of antibodies and pharmaceutical compositions are also provided for treatment of infectious disease, sepsis, and/or autoimmune conditions, and/or for inhibiting an undesirable immune activation that follows gene therapy.
  • Those in need of treatment include those already with cancer as well as those in which the cancer is to be prevented.
  • the mammal to be treated herein may have been diagnosed as having the cancer or may be predisposed or DB2/ 47495312.3 74 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO susceptible to the cancer.
  • treating refers to preventing, delaying the onset of, curing, reversing, attenuating, alleviating, minimizing, suppressing, halting the deleterious effects or stabilizing of discernible symptoms of the above-described cancerous diseases, disorders or conditions. It also includes managing the cancer as described above.
  • manage it is meant reducing the severity of the disease, reducing the frequency of episodes of the disease, reducing the duration of such episodes, reducing the severity of such episodes, slowing/reducing cancer cell growth or proliferation, slowing progression of at least one symptom, amelioration of at least one measurable physical parameter and the like.
  • immunostimulatory anti-PVRIG immune molecules should promote T cell or NK or cytokine immunity against target cells, e.g., cancer, infected or pathogen cells and thereby treat cancer or infectious diseases by depleting the cells involved in the disease condition.
  • immunoinhibitory anti-PVRIG immune molecules should reduce T cell or NK activity and/or or the secretion of proinflammatory cytokines which are involved in the disease pathology of some immune disease such as autoimmune, inflammatory or allergic conditions and thereby treat or ameliorate the disease pathology and tissue destruction that may be associated with such conditions (e.g., joint destruction associated with rheumatoid arthritis conditions).
  • the PVRIG antibodies of the invention, the TIGIT antibodies of the invention, and pembrolizumab are provided in therapeutically effective dosages.
  • a “therapeutically effective dosage” of an anti-PVRIG and or anti-TIGIT immune molecule according to at least some embodiments of the present invention preferably results in a decrease in severity of disease symptoms, an increase in frequency and duration of disease symptom-free periods, an increase in lifespan, disease remission, or a prevention or reduction of impairment or disability due to the disease affliction.
  • a “therapeutically effective dosage” preferably inhibits cell growth or tumor growth by at least about 20%, more preferably by at least about 40%, even more preferably by at least about 60%, and still more preferably by at least about 80% relative to untreated subjects.
  • the ability of a compound to inhibit tumor growth can be evaluated in an animal model system predictive of efficacy in human tumors. Alternatively, this property of a composition can be evaluated by examining the ability of the compound to inhibit, such inhibition in vitro by assays known to the skilled practitioner.
  • a therapeutically effective amount of a therapeutic compound can decrease tumor size, or otherwise ameliorate symptoms in a subject. DB2/ 47495312.3 75 MLB Ref.
  • the “therapeutically effective dosage” of the PVRIG antibody of the invention, the TIGIT antibody of the invention, and pembrolizumab increases the level of serum IFN ⁇ in a subject relative to untreated subjects.
  • the level of serum IFN ⁇ in the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab is increased by at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold, 8-fold, 9-fold, 10-fold, or more, relative to untreated subjects.
  • the level of serum IFN ⁇ in the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab is increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 340%, 350%, 360%, 370%, 380%, 390%, 400%, 410%, 420%, 430%, 440%, 450%, 460%, 470%, 480%, 490%, 500%, 510%, 520%, 530%, 540%, 550%, 560%, 570%, 580%, 590%, 600%, 610%, 620%, 630%, 640%, 650%, 660%, 670%, 680%, 690%, 700%
  • the level of serum IFN ⁇ in the subject is increased by at least 200%, 300%, 400%, or more. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1000%, 1100% 1200%, 1300%, 1400%, 1500% or more. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1200%, or more. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1300%, or more. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1400%, or more. In some embodiments, the level of serum IFN ⁇ in the subject is increased by at least 1500%, or more.
  • the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab exhibits an increased level of serum IFN ⁇ of at least 10 pg/mL, 20 pg/mL, 30 pg/mL, 40 pg/mL, 50 pg/mL, 60 pg/mL, 70 pg/mL, 80 pg/mL, 90 pg/mL, 100 pg/mL, 110 pg/mL, 120 pg/mL, 130 pg/mL, 140 pg/mL, 150 pg/mL, 160 pg/mL, 170 pg/mL, 180 pg/mL, 190 pg/mL, 200 pg/mL, 210 pg/mL, 220 pg/mL, 230 pg/mL, 240 pg/mL, 250 pg/m
  • the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab exhibits an increased level of serum IFN ⁇ of at least 20 pg/mL, 30 pg/mL, 40 pg/mL, or more.
  • the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab exhibits an increased level of serum IFN ⁇ of at least 40 pg/mL, 50 pg/mL, 60 pg/mL, 70 pg/mL, 80 pg/mL or more. In some embodiments, the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab exhibits an increased level of serum IFN ⁇ of at least 70 pg/mL, or more.
  • the subject treated with the therapeutically effective dosage of the PVRIG antibody, the TIGIT antibody, and pembrolizumab exhibits an increased level of serum IFN ⁇ of at least 80 pg/mL, or more.
  • One of ordinary skill in the art would be able to determine a therapeutically effective amount based on such factors as the subject's size, the severity of the subject's symptoms, and the particular composition or route of administration selected. 1. Cancer Treatment [0250]
  • the triple combinations comprising pembrolizumab, a TIGIT antibody, and a PVRIG antibody formulation of the invention find particular use in the treatment of cancer.
  • the antibodies of the invention are immunomodulatory, in that rather than directly attack cancerous cells, the antibodies of the invention stimulate the immune system.
  • cancer immunotherapy is aimed to stimulate the patient’s or subject’s own immune system to eliminate cancer cells, providing long-lived tumor destruction.
  • therapeutic cancer vaccines to induce tumor- specific T cell responses
  • Clinical responses with targeted therapy or conventional anti-cancer therapies tend to be transient as cancer cells develop resistance, and tumor recurrence takes place.
  • Cancer refers broadly to any neoplastic disease (whether invasive or metastatic) characterized by abnormal and uncontrolled cell division causing malignant growth or tumor (e.g., unregulated cell growth).
  • the term “cancer” or “cancerous” as used herein should be understood to encompass any neoplastic disease (whether invasive, non- invasive or metastatic) which is characterized by abnormal and uncontrolled cell division causing malignant growth or tumor, non-limiting examples of which are described herein. This includes any physiological condition in mammals that is typically characterized by unregulated cell growth.
  • triple combinations comprising pembrolizumab, a TIGIT antibody, and a PVRIG antibody formulation of the present invention can be used in the treatment of solid tumors (including, for example, cancers of the lung, liver, breast, brain, GI tract) and blood cancers (including for example, leukemia and preleukemic disorders, lymphoma, plasma cell disorders, including neoplasm) carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies, including advanced cancer forms.
  • the cancer is early.
  • the cancer is advanced (including metastatic).
  • the cancers amenable for treatment of the invention include cancers that express or do not express PVRIG and further include non-metastatic or non-invasive, as well as invasive or metastatic cancers, including cancers where PVRIG expression by immune, stromal, or diseased cells suppresses antitumor responses and anti- invasive immune responses.
  • the anti-PVRIG formulations can be used for the treatment of vascularized tumors.
  • the cancer for treatment using the anti-PVRIG formulations of the present invention includes carcinoma, lymphoma, sarcoma, and/or leukemia.
  • the cancer for treatment using the anti- PVRIG formulations of the present invention includes vascularized tumors, melanoma, non- melanoma skin cancer (squamous and basal cell carcinoma), mesothelioma, squamous cell cancer, lung cancer, small-cell lung cancer, non-small cell lung cancer, neuroendocrine lung cancer (including pleural mesothelioma, neuroendocrine lung carcinoma), NSCL (large cell), DB2/ 47495312.3 78 MLB Ref.
  • melanoma non- melanoma skin cancer (squamous and basal cell carcinoma), mesothelioma, squamous cell cancer, lung cancer, small-cell lung cancer, non-small cell lung cancer, neuroendocrine lung cancer (including pleural mesothelioma, neuroendocrine lung carcinoma), NSCL (large cell), DB2/ 47495312.3 78 MLB Ref.
  • NSCLC non-small cell lung carcinoma
  • NSCLC non-small cell lung carcinoma
  • NSCLC squamous cell soft-tissue sarcoma
  • Kaposi’s sarcoma adenocarcinoma of the lung
  • neuroendocrine lung carcinoma atypical carcinoid lung cancer
  • cancer of the peritoneum including HCC
  • gastric cancer gastric cancer
  • stomach cancer including gastrointestinal cancer
  • pancreatic cancer glioblastoma, cervical cancer, ovarian cancer
  • urothelial cancer bladder cancer
  • hepatoma glioma
  • brain cancer as well as edema, such as that associated with brain tumors
  • breast cancer including, for example, triple negative breast cancer
  • testis cancer testicular germ cell tumor
  • MDS Myelodysplastic syndromes
  • the cancer for treatment using the triple combinations comprising pembrolizumab, a TIGIT antibody, and a PVRIG antibody formulation of the present invention includes a cancer selected from the group consisting of prostate cancer, liver cancer (HCC), rectal cancer, colorectal cancer (CRC), colorectal cancer MSS (MSS- CRC; microsatellite stable colorectal carcinoma/carcinoma), CRC (MSS unknown), metastatic MSS-CRC, refractory metastatic MSS-CRC, MSS-CRC with liver metastasis, ovarian cancer (including ovarian carcinoma), primary peritoneal ovarian carcinoma, advanced epithelial ovarian cancer, advanced epithelial ovarian carcinoma, platinum resistant ovarian cancer, high grade serous adenocarcinoma, endometrial cancer (including endometrial carcinoma), breast cancer, pancreatic cancer, stomach cancer, cervical cancer, head and neck cancer, thyroid cancer, testis cancer, urothelial cancer,
  • HCC liver cancer
  • Cancer therapy herein refers to any method that prevents or treats cancer or ameliorates one or more of the symptoms of cancer.
  • Such therapies will comprise administration of immunostimulatory anti-PVRIG antibodies (including antigen-binding DB2/ 47495312.3 80 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO fragments) either alone or in combination with chemotherapy or radiotherapy or other biologics and for enhancing the activity thereof, i.e., in individuals wherein expression of PVRIG suppresses antitumor responses and the efficacy of chemotherapy or radiotherapy or biologic efficacy.
  • the cancer therapy is a receptor tyrosine kinase inhibitor therapy.
  • the receptor tyrosine kinase inhibitor therapy comprises lenvatinib.
  • the subject administered the anti-PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab is refractory or resistant to treatment with at least two or more prior therapeutic treatments, wherein the prior therapeutic treatments include one or more of fluroropyrimidines (or fluoropyrimidines), irinotecan, oxaliplatin, anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies.
  • the subject administered the anti-PVRIG antibody and anti- TIGIT antibody in combination with pembrolizumab is refractory or resistant to treatment with at least two or more prior therapeutic treatments, wherein the two or more prior therapeutic treatments include fluroropyrimidines (or fluoropyrimidines), irinotecan, and/or oxaliplatin.
  • the subject administered the anti-PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab is refractory or resistant to treatment with no more than three prior therapeutic treatments.
  • the subject administered the anti-PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab is refractory or resistant to treatment with at least three or more prior therapeutic treatments.
  • the subject administered the anti-PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab is refractory or resistant to treatment with at least three or more prior therapeutic treatments, where in the three prior therapeutic treatments include fluroropyrimidines (or fluoropyrimidines), irinotecan, and/or oxaliplatin.
  • the subject administered the anti- PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab has not had prior treatment with an anti-PD-1, anti-PD-L1, or anti-PD-L2-directed therapy.
  • the subject administered the anti-PVRIG antibody and anti-TIGIT antibody in combination with pembrolizumab has not had prior treatment with an immune checkpoint inhibitor, including anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, and/or anti-CTLA4 antibody therapies.
  • DB2/ 47495312.3 81 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO 2.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and v
  • No.114386-5030-WO Compugen Ref.230-WO i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.086.H4(S241P) (SEQ ID NO:34), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CPA.9.086.H4(S241P) (SEQ ID NO:39), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.5
  • the composition has a pH from 6.5 +/- 0.2; and wherein the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.083.H4(S241P) (SEQ ID NO:24), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CPA.9.083.H4(S241P) (SEQ ID NO:29); or i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.086.H4(S241P) (SEQ ID NO:34), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vl
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.7.538.1.
  • a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); (b) from 10 mM to 100 mM histidine; (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0; and wherein the anti-TIGIT antibody comprises: i) a heavy chain variable is from the heavy chain of CPA.9.083.H4(S241P) (SEQ ID NO:24), and ii) a light chain variable domain is from the light chain of CPA.9.083.H4(S241P) (SEQ ID NO:29); or i) a heavy chain variable domain is from the heavy chain of CPA.9.086.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and DB2/ 47495312.3 85 MLB Ref.
  • a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2; and wherein the anti-TIGIT antibody comprises: i) a heavy chain variable is from the heavy chain of CPA.9.083.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: DB2/ 47495312.3 86 MLB Ref.
  • an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH3 region is from
  • No.114386-5030-WO Compugen Ref.230-WO a) a VL-CL, wherein the VL from CPA.9.083.H4(S241P) (SEQ ID NO:29) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.086.H4(S241P) (SEQ ID NO:34) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CPA.9.086.H4(S241P) (SEQ ID NO:39) and wherein the CL region is from human kappa 2 light chain), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg,
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-
  • No.114386-5030-WO Compugen Ref.230-WO a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.538.1.H4(S241P) (SEQ ID NO:19) and wherein the CL region is from human kappa 2 light chain; (b) about 25 mM histidine; (c) about 60 mM NaCl; (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2; and wherein the anti-TIGIT antibody comprises: i) a heavy chain comprising: a
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4(S241
  • No.114386-5030-WO Compugen Ref.230-WO i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti-PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4(S241
  • No.114386-5030-WO Compugen Ref.230-WO i) a heavy chain comprising the heavy chain from CPA.9.083.H4(S241P) (SEQ ID NO:28); and ii) a light chain comprising the light chain from CPA.9.083.H4(S241P) (SEQ ID NO:33); or i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of
  • No.114386-5030-WO Compugen Ref.230-WO (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0; and wherein the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.083.H4(S241P) (SEQ ID NO:24), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CPA.9.083.H4(S241P) (SEQ ID NO:29); or i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.086.H4(S241P)
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of
  • No.114386-5030-WO Compugen Ref.230-WO i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19).
  • the anti-TIGIT antibody comprises: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.083.H4(S241P) (SEQ ID NO:24), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of CPA.9.083.H4(S241P) (SEQ ID NO:29); or i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CPA.9.086.H4(S241P
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 DB2/ 47495312.3 94 MLB Ref.
  • the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:14), and ii) a light chain variable domain is from the light chain of CHA.7.538.1.H4(S241P) (SEQ ID NO:19); (b) from 10 mM to 100 mM histidine; (c)
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a)
  • the composition has a pH from 5.5 to 7.0; and wherein the anti-TIGIT antibody comprises: a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.083.H4(S241P) (SEQ ID NO:24) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CPA.9.083.H4(S241P) (SEQ ID NO:29) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.086.H4(S241P) (SEQ ID NO:34) and wherein the CH
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: DB2/ 47495312.3 98 MLB Ref.
  • No.114386-5030-WO Compugen Ref.230-WO a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a
  • No.114386-5030-WO Compugen Ref.230-WO a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.086.H4(S241P) (SEQ ID NO:34) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CPA.9.086.H4(S241P) (SEQ ID NO:39) and wherein the CL region is from human kappa 2 light chain), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4
  • the anti-TIGIT antibody comprises: i) a heavy chain comprising the heavy chain from CPA.9.083.H4(S241P) (SEQ ID NO:28); and ii) a light chain comprising the light chain from CPA.9.083.H4(S241P) (SEQ ID NO:33); or i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4
  • No.114386-5030-WO Compugen Ref.230-WO (d) about 100 mM L-Arginine; and (e) about 0.01% % w/v polysorbate 80, wherein the composition has a pH from 6.5 +/- 0.2; and wherein the anti-TIGIT antibody comprises: i) a heavy chain comprising the heavy chain from CPA.9.083.H4(S241P) (SEQ ID NO:28); and ii) a light chain comprising the light chain from CPA.9.083.H4(S241P) (SEQ ID NO:33); or i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain comprising the vhCDR1, vhCDR2, and vhCDR3 from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain comprising the vlCDR1, vlCDR2, and vlCDR3 from the light chain of
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: DB2/ 47495312.3 103 MLB Ref.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain variable domain is from the heavy chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:4), and ii) a light chain variable domain is from the light chain of CHA.7.518.1.H4(S241P) (SEQ ID NO:9); or i) a heavy chain variable domain is from the heavy chain of CHA.
  • a heavy chain variable domain is from the heavy chain of CPA.9.086.H4(S241P) (SEQ ID NO:34), and ii) a light chain variable domain is from the light chain of CPA.9.086.H4(S241P) (SEQ ID NO:39) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg of the anti-TIGIT antibody.
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a)
  • No.114386-5030-WO Compugen Ref.230-WO (c) from 30 mM to 100 mM NaCl; (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0; and wherein the anti-TIGIT antibody comprises: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.083.H4(S241P) (SEQ ID NO:24) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CPA.9.083.H4(S241P) (SEQ ID NO:29) and wherein the CL region is from human kappa 2 light chain; or i)
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: DB2/ 47495312.3 108 MLB Ref.
  • an anti-PVRIG antibody comprising: i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.518.1.H4(S241P) (SEQ ID NO:4) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CHA.7.518.1.H4(S241P) (SEQ ID NO:9) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CHA.7.538.1.H4(S241P) (SEQ ID NO:14) and wherein the CH1- hinge-CH2-CH3 region is from
  • No.114386-5030-WO Compugen Ref.230-WO a) a VL-CL, wherein the VL from CPA.9.083.H4(S241P) (SEQ ID NO:29) and wherein the CL region is from human kappa 2 light chain; or i) a heavy chain comprising: a) a VH-CH1-hinge-CH2-CH3, wherein the VH is from CPA.9.086.H4(S241P) (SEQ ID NO:34) and wherein the CH1-hinge- CH2-CH3 region is from IgG4; and ii) a light chain comprising: a) a VL-CL, wherein the VL from CPA.9.086.H4(S241P) (SEQ ID NO:39) and wherein the CL region is from human kappa 2 light chain), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg,
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4
  • No.114386-5030-WO Compugen Ref.230-WO (d) from 20 mM to 150 mM L-Arginine; and (e) from 0.005% to 0.1% w/v polysorbate 80, wherein the composition has a pH from 5.5 to 7.0; and wherein the anti-TIGIT antibody comprises: i) a heavy chain comprising the heavy chain from CPA.9.083.H4(S241P) (SEQ ID NO:28); and ii) a light chain comprising the light chain from CPA.9.083.H4(S241P) (SEQ ID NO:33); or i) a heavy chain comprising the heavy chain from CPA.9.086.H4(S241P) (SEQ ID NO:38); and ii) a light chain comprising the light chain from CPA.9.086.H4(S241P) (SEQ ID NO:43) ), and wherein the anti-TIGIT antibody is administered at a dosage of about 0.01 mg
  • the present invention provides for treatment of cancer in a subject in need thereof by administration of 200 mg/kg pembrolizumab, an anti-TIGIT, and a stable liquid pharmaceutical formulation of an anti-PVRIG antibody, wherein the anti- PVRIG antibody is administered at a dosage of about 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg, and wherein the stable liquid formulation of the anti-PVRIG antibody comprises: (a) an anti-PVRIG antibody comprising: i) a heavy chain comprising the heavy chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:8); and ii) a light chain comprising the light chain from CHA.7.518.1.H4(S241P) (SEQ ID NO:13); or i) a heavy chain comprising the heavy chain from CHA.7.538.1.H4
  • the pembrolizumab is administered as a dosage of about 200 mg/kg. In some embodiments, the pembrolizumab is administered as a dosage of about 200 mg/kg every 3 weeks. [0282] In some embodiments, pembrolizumab is administered as a dosage of about 2 mg/kg to 10 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 2 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 2 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 3 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 4 mg/kg.
  • pembrolizumab is administered as a dosage of about 5 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 6 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 7 mg/kg. In some DB2/ 47495312.3 112 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO embodiments, pembrolizumab is administered as a dosage of about 8 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 9 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 10 mg/kg.
  • pembrolizumab is administered as a dosage of about no more than 2 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 1 mg/kg to 2 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 0.1 mg/kg to 1 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 0.01 mg/kg to 0.1 mg/kg. [0284] In some embodiments, pembrolizumab is administered as a dosage of about at least 10 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 10 mg/kg to 20 mg/kg.
  • pembrolizumab is administered as a dosage of about 20 mg/kg to 30 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 30 mg/kg to 40 mg/kg. In some embodiments, pembrolizumab is administered as a dosage of about 40 mg/kg to 50 mg/kg. [0285] In some embodiments, pembrolizumab is administered about every 1 week to every 6 weeks. In some embodiments, pembrolizumab is administered about every week. In some embodiments, pembrolizumab is administered about every 2 weeks. In some embodiments, pembrolizumab is administered about every 3 weeks. In some embodiments, pembrolizumab is administered about every 4 weeks.
  • pembrolizumab is administered about every 5 weeks. In some embodiments, pembrolizumab is administered about every 6 weeks. [0286] In some embodiments, pembrolizumab is administered as a dosage of about 2 mg/kg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 10 mg/kg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 200 mg every 3 weeks. In some embodiments, pembrolizumab is administered as a dosage of about 400 mg every 6 weeks.
  • the pembrolizumab is administered over about 10 minutes, over about 15 minutes, over about 20 minutes, over about 25 minutes, over about 30 minutes, over about 35 minutes, or over about 40 minutes. In some embodiments, the pembrolizumab is administered over about 30 minutes +/- 10 minutes. [0288] Further disclosures of pembrolizumab are provided in https://www.accessdata.fda.gov/spl/data/157262d6-15e0-4b0a-968f- DB2/ 47495312.3 113 MLB Ref.
  • the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered in any order. In some embodiments, the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered simultaneously. In some embodiments, the anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab are administered sequentially.
  • the order of administration is anti-PVRIG antibody, anti-TIGIT antibody, and pembrolizumab. In some embodiments, the order of administration is anti-TIGIT antibody, anti-PVRIG antibody, and pembrolizumab. In some embodiments, the order of administration is pembrolizumab, anti-PVRIG antibody, and anti- TIGIT antibody. In some embodiments, the order of administration is pembrolizumab, anti- TIGIT antibody, and anti-PVRIG antibody. In some embodiments, the order of administration is anti-PVRIG antibody, pembrolizumab, and anti-TIGIT antibody. In some embodiments, the order of administration is anti-TIGIT antibody, pembrolizumab, and anti- PVRIG antibody.
  • EXAMPLE 1 PVRIG ANTIBODY FORMULATION TESTING
  • the PVRIG antibody that was formulated in each buffer (A and B) was spiked with the corresponding excipients to create the 20 conditions listed in Figure 6. Each formulation was referred to by the Formulation ID.
  • the formulated material was subjected to the stress and storage conditions in Figures 7A-7B.
  • Formulation Study Procedural Steps Sampling Requirements [0292] For each condition, a total of two vials were required for testing with one additional spare vial.
  • Samples were assayed as described in this example. 2–8 °C Storage for 8 weeks (Testing at 0, 2, 4 and 8 weeks) [0299] Twelve vials were taken from each of the 20 formulations (a total of 240 vials including T0 vials) and stored at 2–8 °C. Two vials at T0 were analyzed immediately for MFI and appearance. All other samples were labeled with temperature and time point and stored frozen at ⁇ -60 °C until analysis. Samples were brought to room temperature prior to analysis.
  • Samples were brought to room temperature prior to analysis. [0307] Samples were assayed as described in this example. Testing Plan and Schedule [0308] For each formulation condition, sample and test were performed. Results [0309] Each formulation was evaluated and compared to the conditions studied. [0310] The critical assay results were compiled and analyzed. The critical assays analyzed to determine an appropriate formulation were SEC, cIEF and MFI. SEC high molecular and low molecular weight species were monitored throughout the study. cIEF results were obtained throughout the study. Finally, MFI particles/mL throughout the various sizes were monitored.
  • EXAMPLE 2 PVRIG ANTIBODY FORMULATION DESCRIPTION AND COMPOSITION OF SOLUTION FOR INFUSION
  • the formulation is provided as a sterile preservative free liquid dosage form at a concentration of 20 mg/mL in a 10R Type I clear borosilicate glass vial equipped with a gray bromobutyl rubber stopper and aluminum flip cap crimp. The vials are filled to a target volume of 10 mL. The formulation is stored and shipped frozen at -20 C. Prior to use, the DB2/ 47495312.3 116 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO vials are thawed at ambient temperature, mixed by gentle swirling.
  • the formulation was produced by thawing and pooling the Drug Substance, followed by 0.22 ⁇ m sterile filtration and filling into sterile 10R glass vials at Vetter.
  • the formulation components and quantitative composition of the drug product on a nominal amount per vial unit (10 mL) is presented in the Table 1 below.
  • Table 1 Composition [0315] A sufficient volume is filled into vials based on the net fill weight to ensure a withdrawable volume of 10 mL.
  • EXAMPLE 3 A PHASE 1 STUDY OF THE SAFETY AND TOLERABILITY OF CPA.9.086.H4(S241P) IN SUBJECTS WITH ADVANCED MALIGNANCIES.
  • Arm 2 Combination Product: Evaluation of safety/tolerability: CPA.9.086.H4(S241P) in combination with CHA.7.518.1.H4(S241P) (both at the RDFE). Both study drugs will be evaluated at the RDFE for assessment of safety and tolerability. All study drugs will be administered IV every 3 weeks.
  • CPA.9.086.H4(S241P) will be administered IV every 3 weeks.
  • Arm 4 CPA.9.086.H4(S241P) in combination with CHA.7.518.1.H4(S241P) (both at the RDFE).
  • MSS-CRC For Triplet combination MSS-CRC: [0327] Histologically confirmed adenocarcinoma of the colon/rectum [0328] Stage IV disease [0329] MSS-CRC status by an FDA approved test [0330] Disease progression with no more than 3 prior lines of treatment including fluroropyrimidines (or fluoropyrimidines), irinotecan, and oxaliplatin or more than 3 prior lines of treatment including fluroropyrimidines (or fluoropyrimidines), irinotecan, and oxaliplatin. Key Exclusion Criteria: [0331] Prior treatment with a TIGIT inhibitor.
  • EXAMPLE 4 CRC (MSS) TRIPLET EXPANSION COHORT: CPA.9.086.H4(S241P) + CHA.7.518.1.H4(S241P) + PEMBROLIZUMAB Exclusion criterion [0358] Prior treatment with an anti-PD-1/PD-L1/PD-L2, anti-CD96 antibody, anti-OX-40 antibody, anti-CD137 antibody, anti-LAG3, anti-TIM3, or anti-CTLA4 antibody.
  • EXAMPLE 5 CLINICAL STUDY OF OVARIAN CANCER (TRIPLET EXPANSION) Inclusion criteria [0359] Subjects with advanced epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal carcinoma are eligible.
  • Platinum resistant ovarian cancer defined as disease recurrence ⁇ 6 months after completion of a platinum-containing regimen. Subjects with primary platinum DB2/ 47495312.3 121 MLB Ref. No.114386-5030-WO Compugen Ref.230-WO refractory disease are ineligible. Primary platinum refractory disease is defined as progression of disease prior to completion of 1st line platinum therapy or immediately following ( ⁇ 3 months following last date of chemotherapy). [0361] Received ⁇ 3 prior lines for PROC; maintenance bevacizumab or PARP are not included as a line of therapy. [0362] Subjects who have received PARP inhibitor therapy are eligible .
  • CHA.7.518.1.H4(S241P) + nivolumab in patients with MSS-CRC and liver metastases [12/22(77%)], [ORR 2/17 (12%); disease control rate [DCR] 5/17 (29%)] [1].
  • CHA.7.518.1.H4(S241P) is a novel, 1st-in-class immune checkpoint inhibitor (ICI) that binds to PVRIG, a DNAM-1 axis member, leading to activation of T and NK-cells;
  • CPA.9.086.H4(S241P) is an ICI of TIGIT.
  • Pembrolizumab is an ICI of PD-1.

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Abstract

La présente invention concerne des polythérapies pour le traitement du cancer réfractaire ou résistant à au moins deux traitements thérapeutiques antérieurs, le traitement comprenant des anticorps anti-PVRIG, des anticorps anti-TIGIT et du pembrolizumab, y compris des formulations pharmaceutiques liquides stables de ceux-ci, en particulier des anticorps anti-PVRIG.
EP24714367.0A 2023-02-27 2024-02-27 Polythérapie triple avec des anticorps anti-pvrig, des anticorps anti-tigit et du pembrolizumab Pending EP4673225A2 (fr)

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US202363487230P 2023-02-27 2023-02-27
US202363489119P 2023-03-08 2023-03-08
PCT/US2024/017551 WO2024182443A2 (fr) 2023-02-27 2024-02-27 Polythérapie triple avec des anticorps anti-pvrig, des anticorps anti-tigit et du pembrolizumab

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EP4673225A2 true EP4673225A2 (fr) 2026-01-07

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WO2024182443A2 (fr) 2024-09-06

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