# US 10,870,703 B2 · Methods of treating immunotherapy-related toxicity using a GM-CSF antagonist > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,870,703 B2 - Document ID: US10870703B2 - Status: Granted - Kind: B2 - Application number: 16/149,346 - Filed: 2018-10-02 - Published: 2019-06-27 - Granted: 2020-12-22 - Earliest claimed: 2017-10-02 - Assignees: Humanigen, Inc. - Applicants: Humanigen, Inc. - Inventors: Cameron Durrant; Dale Chappell - Web page: https://patentgrep.com/patent/US10870703B2 ## CPC - C07K 16/2893: {against CD52} - A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines} - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/4211: {CD19 or B4} - A61K 40/4217: {Receptors for interleukins [IL]} - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF - C07K 16/243: {Colony Stimulating Factors} - C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex} - C07K 16/2887: {against CD20} - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - A61K 2039/505: {comprising antibodies} - A61K 2239/31: characterized by the route of administration - A61K 2239/38: characterised by the dose, timing or administration schedule - A61K 2239/48: Blood cells, e.g. leukemia or lymphoma - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/54: F(ab')2 - C07K 2317/55: Fab or Fab' - C07K 2317/565: Complementarity determining region [CDR] - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/626: Diabody or triabody - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract Methods of inhibiting or reducing the incidence or the severity of immunotherapy-related toxicity in a subject, the method comprising a step of administering a recombinant hGM-CSF antagonist to the subject, wherein said administering inhibits or reduces the incidence or the severity of immunotherapy-related toxicity in said subject, are provided. An hGM-CSF antagonist for use in methods of inhibiting or reducing the incidence or the severity of immunotherapy-related toxicity in a subject also are provided. ## Claims ### Claim 1 A method treating cytokine release syndrome (CRS) in a subject treated with CAR-T cell therapy, the method comprising a step of administering a recombinant hGM-CSF antagonist to the subject, wherein the recombinant hGM-CSF antagonist is anti-hGM-CSF antibody lenzilumab and wherein said administering neutralizes hGM-CSF in said subject. ### Claim 2 (depends on 1) The method of claim 1, wherein said treating the CRS comprises reducing a concentration of at least one inflammation-associated factor in the serum, tissue fluid, or in the CSF of said subject. ### Claim 3 (depends on 2) The method of claim 2, wherein said inflammation-associated factor is selected from the group consisting of: C-reactive protein, hGM-CSF, sIL2Rα, IL-5, IL-6, IL-8, IP10, IL-10, IL-15, MCP-1, MIG, MIP1β, IFNγ, CX3CR1, TNFα, and any combination thereof. ### Claim 4 (depends on 1) The method of claim 1, wherein administration of said recombinant hGM-CSF antagonist does not reduce the efficacy of said CAR-T cell therapy. ### Claim 5 (depends on 1) The method of claim 1, wherein said CAR-T cell therapy is administered at higher doses than it would be administered without the administration of said hGM-CSF antagonist. ### Claim 6 (depends on 1) The method of claim 1, wherein administration of said recombinant hGM-CSF antagonist occurs prior to, concurrent with, or following said CAR-T cell therapy. ### Claim 7 (depends on 1) The method of claim 1, wherein said recombinant hGM-CSF antagonist is co-administered with corticosteroids, anti-IL-6 antibodies, tocilizumab, cyclosporine, antiepileptics, benzodiazepines, acetazolamide, hyperventilation therapy, or hyperosmolar therapy, or any combination thereof. ### Claim 8 (depends on 1) The method of claim 1, wherein said CRS toxicity comprises a brain disease, damage or malfunction. ### Claim 9 (depends on 8) The method of claim 8, wherein said brain disease, damage or malfunction comprises CAR-T cell related neurotoxicity or CAR-T cell related encephalopathy syndrome (CRES). ### Claim 10 (depends on 9) The method of claim 9, wherein the CAR-T cell related neurotoxicity is reduced by about 90% compared to a reduction in neurotoxicity in a subject treated with CAR-T cells and a control antibody. ### Claim 11 (depends on 8) The method of claim 8, wherein said treating the CRS comprises reducing headaches, delirium, anxiety, tremor, seizure activity, confusion, alterations in wakefulness, hallucinations, dysphasia, ataxia, apraxia, facial nerve palsy, motor weakness, seizures, nonconvulsive EEG seizures, altered levels of consciousness, coma, endothelial activation, vascular leak, intravascular coagulation, or any combination thereof in said subject. ### Claim 12 (depends on 8) The method of claim 8, wherein the serum concentration of ANG2 or VWF, or the serum ANG2:ANG1 ratio of said subject is reduced. ### Claim 13 (depends on 8) The method of claim 8, wherein said subject has a body temperature above 38° C., IL6 serum concentration above 16 pg/ml, or MCP-1 serum concentration above 1,300 pg/ml during the first 36 hours after infusion of said CAR-T cells. ### Claim 14 (depends on 8) The method of claim 8, wherein said subject is predisposed to have said brain disease, damage or malfunction. ### Claim 15 (depends on 8) The method of claim 8, wherein said subject has an ANG2:ANG1 ratio in serum above 1 prior to the infusion of said CAR-T cells. ### Claim 16 (depends on 1) The method of claim 1, wherein said CRS comprises hemophagocytic lymphohistiocytosis (HLH) or macrophage-activation syndrome (MAS). ### Claim 17 (depends on 16) The method of claim 16, wherein said treating the CRS inhibits or reduces HLH or MAS and comprises increasing survival time and/or time to relapse, reducing macrophage activation, reducing the concentration of circulating IFNy, or reducing the concentration of circulating of hGM-CSF, or any combination thereof. ### Claim 18 (depends on 16) The method of claim 16, wherein said subject presents with fever, splenomegaly, cytopenias involving two or more lines, hypertriglyceridemia, hypofibrinogenemia, hemophagocytosis, low or absent NK-cell activity, ferritin serum concentration above 500 U/ml, soluble CD25 serum concentration above 2400 U/ml, or any combination thereof. ### Claim 19 (depends on 16) The method of claim 16, wherein said subject is predisposed to acquiring HLH or MAS. ### Claim 20 (depends on 16) The method of claim 16, wherein said subject carries a mutation in a gene selected from: PRF1, UNC13D, STX11, STXBP2, or RAB27A, or has reduced expression of perforin, or any combination thereof. ### Claim 21 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody blocks binding of hGM-CSF to the alpha subunit of the hGM-CSF receptor. ### Claim 22 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody has an affinity ranging from about 5 pM to about 50 pM. ### Claim 23 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody lenzilumab comprises an engineered human variable region. ### Claim 24 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody lenzilumab binds to the same epitope as chimeric 19/2. ### Claim 25 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody lenzilumab comprises a VH region that comprises a CDR3 binding specificity determinant RQRFPY (SEQ ID NO: 12), a J segment, and a V-segment, wherein the J-segment comprises at least 95% identity to human JH4 (YFDYWGQGTLVTVSS) (SEQ ID NO: 14) and the V-segment comprises at least 90% identity to the human germ line VH1 1-03 sequence. ### Claim 26 (depends on 25) The method of claim 25, wherein the J segment comprises YFDYWGQGTLVTVSS (SEQ ID NO: 14). ### Claim 27 (depends on 25) The method of claim 25, wherein the CDR3 comprises RQRFPYYFDY (SEQ ID NO: 15). ### Claim 28 (depends on 25) The method of claim 25, wherein the anti-hGM-CSF antibody comprises a VH CDR1 and a VH CDR2 as shown in the VH region having the sequence of VH #5 set forth in FIG. 1. ### Claim 29 (depends on 25) The method of claim 25, wherein the V-segment sequence has a VH V segment sequence of VH #5 shown in FIG. 1. ### Claim 30 (depends on 25) The method of claim 25, wherein the VH has the sequence of VH #5 set forth in FIG. 1. ### Claim 31 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody comprises a VL-region that comprises a CDR3 comprising the amino acid sequence FNK. ### Claim 32 (depends on 31) The method of claim 31, wherein the anti-hGM-CSF antibody comprises a VL region that comprises a CDR3 comprising QQFNKSPLT (SEQ ID NO: 18). ### Claim 33 (depends on 31) The method of claim 31, wherein the VL region comprises a V segment that has at least 95% identity to the VKIII A27 V-segment sequence as shown in FIG. 1. ### Claim 34 (depends on 31) The method of claim 31, wherein the VL region has the sequence VK #2 set forth in FIG. 1. ### Claim 35 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody has a VH region CDR3 binding specificity determinant RQRFPY (SEQ ID NO: 12) and a VL region that has a CDR3 comprising QQFNKSPLT (SEQ ID NO: 18). ### Claim 36 (depends on 1) The method of claim 1, wherein the anti-hGM-CSF antibody lenzilumab has a VH region having the sequence VH #5 set forth in FIG. 1 and a VL region having the sequence VK #2 set forth in FIG. 1. ### Claim 37 (depends on 36) The method of claim 36, wherein the VH region, the VL region, or both the VH and VL region amino acid sequences comprise a methionine at the N-terminus. ### Claim 38 A method of inhibiting or reducing the incidence or the severity of CAR-T related toxicity in a subject, the method comprising a step of administering a recombinant hGM-CSF antagonist to the subject, wherein the recombinant hGM-CSF antagonist is anti-hGM-CSF antibody lenzilumab and wherein said administering inhibits or reduces the incidence or the severity of CAR-T related toxicity in said subject. ### Claim 39 (depends on 38) The method of claim 38, wherein said CAR-T related toxicity comprises neurotoxicity, CRS, or a combination thereof. ### Claim 40 (depends on 39) The method of claim 39, wherein the CAR-T cell related neurotoxicity is reduced by about 50% compared to a reduction in neurotoxicity in a subject treated with CAR-T cells and a control antibody. ### Claim 41 (depends on 39) The method of claim 39, wherein said inhibiting or reducing incidence of CRS comprises increasing survival time and/or time to relapse, reducing macrophage activation, or reducing the concentration of circulating hGM-CSF, or any combination thereof. ### Claim 42 (depends on 38) The method of claim 38, wherein said subject presents with fever (with or without rigors, malaise, fatigue, anorexia, myalgia, arthralgia, nausea, vomiting, headache, skin rash, diarrhea, tachypnea, hypoxemia, hypoxia, shock, cardiovascular tachycardia, widened pulse pressure, hypotension, capillary leak, increased early cardiac output, diminished late cardiac output, elevated D-dimer, hypofibrinogenemia with or without bleeding, azotemia, transaminitis, hyperbilirubinemia, mental status changes, confusion, delirium, frank aphasia, hallucinations, tremor, dysmetria, altered gait, seizures, organ failure, or any combination thereof. ### Claim 43 (depends on 38) The method of claim 38, wherein the inhibiting or reducing the incidence or the severity of CAR-T related toxicity comprises preventing the onset of CAR-T related toxicity. ### Claim 44 (depends on 1) The method of claim 1, wherein the subject is a human. ### Claim 45 A method of treating CAR-T cell related neurotoxicity in a subject treated with CAR-T cell therapy, the method comprising a step of administering a recombinant hGM-CSF antagonist to the subject, wherein the recombinant hGMCSF antagonist is anti-hGM-CSF antibody lenzilumab and wherein said administering neutralizes hGM-CSF in said subject. ### Claim 46 (depends on 45) The method of claim 45, wherein said treating the CART-cell related neurotoxicity comprises reducing a concentration of at least one inflammation-associated factor in the serum, tissue fluid, or in the CSF of said subject. ### Claim 47 (depends on 46) The method of claim 46, wherein said inflammation-associated factor is selected from the group consisting of: C-reactive protein, hGM-CSF, sIL2Rα, IL-5, IL-6, IL-8, IP10, IL-10, IL-15, MCP-1, MIG, MIP1β, IFNγ, CX3CR1, TNFα, and any combination thereof. ### Claim 48 (depends on 45) The method of claim 45, wherein administration of said recombinant hGM-CSF antagonist lenzilumab does not reduce the efficacy of said CAR-T cell therapy. ### Claim 49 (depends on 45) The method of claim 45, wherein said CAR-T cell therapy is administered at higher doses than it would be administered without the administration of said hGM-CSF antagonist. ### Claim 50 (depends on 45) The method of claim 45, wherein administration of said recombinant hGM-CSF antagonist occurs prior to, concurrent with, or following said CAR-T cell therapy. ### Claim 51 (depends on 45) The method of claim 45, wherein said recombinant hGM-CSF antagonist is coadministered with corticosteroids, anti-IL-6 antibodies, tocilizumab, cyclosporine, antiepileptics, benzodiazepines, acetazolamide, hyperventilation therapy, hyperosmolar therapy, or any combination thereof. ### Claim 52 (depends on 45) The method of claim 45, wherein said CAR-T cell related neurotoxicity comprises a brain disease, damage or malfunction. ### Claim 53 (depends on 52) The method of claim 52, wherein said brain disease, damage or malfunction comprises CAR-T cell related encephalopathy syndrome (CRES). ### Claim 54 (depends on 52) The method of claim 52, wherein the CAR-T cell related neurotoxicity is reduced by about 90% compared to a reduction in neurotoxicity in a subject treated with CAR-T cells and a control antibody. ### Claim 55 (depends on 52) The method of claim 52, wherein said treating the CART-cell related neurotoxicity comprises reducing headaches, delirium, anxiety, tremor, seizure activity, confusion, alterations in wakefulness, hallucinations, dysphasia, ataxia, apraxia, facial nerve palsy, motor weakness, seizures, nonconvulsive EEG seizures, altered levels of consciousness, coma, endothelial activation, vascular leak, intravascular coagulation, or any combination thereof in said subject. ### Claim 56 (depends on 52) The method of claim 52, wherein the serum concentration of ANG2 or VWF, or the serum ANG2:ANG1 ratio of said subject is reduced. ### Claim 57 (depends on 52) The method of claim 52, wherein said subject has a body temperature above 38° C., IL6 serum concentration above 16 pg/ml, or MCP-1 serum concentration above 1,300 pg/ml during the first 36 hours after infusion of said CAR-T cells. ### Claim 58 (depends on 52) The method of claim 52, wherein said subject is predisposed to have said brain disease, damage or malfunction. ### Claim 59 (depends on 52) The method of claim 52, wherein said subject has an ANG2:ANG1 ratio in serum above 1 prior to the infusion of said CAR-T cells. ### Claim 60 (depends on 45) The method of claim 45, wherein said CART-cell related neurotoxicity comprises hemophagocytic lymphohistiocytosis (HLH) or macrophage-activation syndrome (MAS). ### Claim 61 (depends on 60) The method of claim 60, wherein said treating CAR-T cell related neurotoxicity inhibits or reduces HLH or MAS and comprises increasing survival time and/or time to relapse, reducing macrophage activation, reducing the concentration of circulating IFNy, or reducing the concentration of circulating of hGM-CSF, or any combination thereof. ### Claim 62 (depends on 60) The method of claim 60, wherein said subject presents with fever, splenomegaly, cytopenias involving two or more lines, hypertriglyceridemia, hypofibrinogenemia, hemophagocytosis, low or absent NK-cell activity, ferritin serum concentration above 500 U/ml, soluble CD25 serum concentration above 2400 U/ml, or any combination thereof. ### Claim 63 (depends on 60) The method of claim 60, wherein said subject is predisposed to acquiring HLH or MAS. ### Claim 64 (depends on 60) The method of claim 60, wherein said subject carries a mutation in a gene selected from: PRF1, UNC13D, STX11, STXBP2, or RAB27A, or has reduced expression of perforin, or any combination thereof. ### Claim 65 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody lenzilumab blocks binding of hGM-CSF to the alpha subunit of the hGM-CSF receptor. ### Claim 66 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody has an affinity ranging from about 5 pM to about 50 pM. ### Claim 67 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody lenzilumab comprises an engineered human variable region. ### Claim 68 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody lenzilumab binds to the same epitope as chimeric 19/2. ### Claim 69 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody lenzilumab comprises a VH region that comprises a CDR3 binding specificity determinant RQRFPY (SEQ ID NO: 12), a J segment, and a V-segment, wherein the J-segment comprises at least 95% identity to human JH4 (YFDYWGQGTLVTVSS) (SEQ ID NO: 14) and the V-segment comprises at least 90% identity to a human germ line VH1 1-03 sequence. ### Claim 70 (depends on 69) The method of claim 69, wherein the J segment comprises YFDYWGQGTLVTVSS (SEQ ID NO: 14). ### Claim 71 (depends on 69) The method of claim 69, wherein the CDR3 comprises RQRFPYYFDY (SEQ ID NO: 15). ### Claim 72 (depends on 69) The method of claim 69, wherein the anti-hGM-CSF antibody comprises a VH CDR1 and a VH CDR2 as shown in a VH region having the sequence of VH #5 set forth in FIG. 1. ### Claim 73 (depends on 69) The method of claim 69, wherein the V-segment sequence has a VH V segment sequence of VH #5 shown in FIG. 1. ### Claim 74 (depends on 69) The method of claim 69, wherein the VH has the sequence of VH #5 set forth in FIG. 1. ### Claim 75 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody comprises a VL region that comprises a CDR3 comprising the amino acid sequence FNK. ### Claim 76 (depends on 75) The method of claim 75, wherein the anti-hGM-CSF antibody comprises a VL region that comprises a CDR3 comprising QQFNKSPLT (SEQ ID NO: 18). ### Claim 77 (depends on 75) The method of claim 75, wherein the VL region comprises a CDR1 and CDR2 of a VL region having the sequence of VK #2 shown in FIG. 1. ### Claim 78 (depends on 75) The method of claim 75, wherein the VL region comprises a V segment that has at least 95% identity to the VKIII A27 V-segment sequence as shown in FIG. 1. ### Claim 79 (depends on 75) The method of claim 75, wherein the VL region has the sequence of VK #2 set forth in FIG. 1. ### Claim 80 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody has a VH region CDR3 binding specificity determinant RQRFPY (SEQ ID NO: 12) and a VL region that has a CDR3 comprising QQFNKSPLT (SEQ ID NO: 18). ### Claim 81 (depends on 45) The method of claim 45, wherein the anti-hGM-CSF antibody lenzilumab has a VH region having sequence VH #5 set forth in FIG. 1 and a VL region having sequence VK #2 set forth in FIG. 1. ### Claim 82 (depends on 81) The method of claim 81, wherein the VH region, the VL region, or both the VH and VL region amino acid sequences comprise a methionine at the N-terminus.