# US 11,440,966 B2 · Multispecific antibodies against CD40 and CD137 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,440,966 B2 - Document ID: US11440966B2 - Status: Granted - Kind: B2 - Application number: 17/345,628 - Filed: 2021-06-11 - Published: 2021-10-14 - Granted: 2022-09-13 - Earliest claimed: 2016-07-14 - Assignees: Genmab A/S; BioNTech SE - Applicants: Genmab A/S; BioNTech SE - Inventors: Isil Altintas; David Satijn; Rik Rademaker; Paul Parren; Friederike Gieseke; Ugur Sahin - Web page: https://patentgrep.com/patent/US11440966B2 ## CPC - C07K 16/2878: {against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95} - A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines} - A61P 31/04: Antibacterial agents - A61P 35/00: Antineoplastic agents - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - G01N 33/56972: {White blood cells} - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/524: CH2 domain - C07K 2317/526: CH3 domain - C07K 2317/53: Hinge - C07K 2317/565: Complementarity determining region [CDR] - C07K 2317/75: Agonist effect on antigen - 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 - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Multispecific antibodies binding to human CD40 and human CD137, methods for preparing such multispecific antibodies, and methods of using such multispecific antibodies for therapeutic or other purposes. ## Claims ### Claim 1 A multispecific antibody or a fragment thereof comprising a first heavy chain and a first light chain comprising a first antigen-binding region capable of binding to human CD40 and a second heavy chain and a second light chain comprising a second antigen-binding region capable of binding to human CD137 wherein the first antigen-binding region capable of binding to human CD40 comprises a first heavy chain variable (VH) region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 1, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 2, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 4, a LCDR2 sequence comprising the amino acid sequence YTS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 5; wherein the second antigen-binding region capable of binding to human CD137 comprises a second VH region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 64, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 65, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 66, and a second VL region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 67, a LCDR2 sequence comprising the amino acid sequence GAS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 68, wherein in at least one of said first and second heavy chains one or more amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively. ### Claim 2 (depends on 1) The multispecific antibody or fragment thereof of claim 1, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to EU numbering are F, E, and A, respectively, in said first and second heavy chains. ### Claim 3 (depends on 2) The multispecific antibody or fragment thereof of claim 2, wherein the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to EU numbering of both the first heavy chain and the second heavy chain are F, E, and A, respectively, and wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering of the first heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering of the second heavy chain is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering of the first heavy chain is R, and the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering of the second heavy chain is L. ### Claim 4 (depends on 1) The multispecific antibody or fragment thereof of claim 1, wherein said antibody is able to cross-link a first cell expressing human CD40 and a second cell expressing human CD137. ### Claim 5 (depends on 1) The multispecific antibody or fragment thereof of claim 1, wherein said antibody is a bispecific antibody. ### Claim 6 (depends on 1) The multispecific antibody or fragment thereof of claim 1, wherein (i) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said second heavy chain, or (ii) the amino acid in the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said first heavy chain, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said second heavy chain. ### Claim 7 (depends on 1) The multispecific antibody or fragment thereof according to claim 1, wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117. ### Claim 8 (depends on 1) The multispecific antibody or fragment thereof according to claim 1, wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123. ### Claim 9 (depends on 1) The multispecific antibody or fragment thereof according to claim 1, wherein the first VL region comprises the amino acid sequence as set forth in SEQ ID NO:121. ### Claim 10 (depends on 1) The multispecific antibody or fragment thereof according to claim 1, wherein the second VL region comprises the amino acid sequence as set forth in SEQ ID NO: 127. ### Claim 11 A multispecific antibody or a fragment thereof comprising a first heavy chain and a first light chain comprising a first antigen-binding region capable of binding to human CD40 and a second heavy chain and a second light chain comprising a second antigen-binding region capable of binding to human CD137, wherein the first antigen-binding region comprises a first heavy chain variable (VH) region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 1, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 2, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 4, a LCDR2 sequence comprising the amino acid sequence YTS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 5; wherein the second antigen-binding region comprises a second VH region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 64, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 65, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 66, and a second VL region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 67, a LCDR2 sequence comprising the amino acid sequence GAS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 68, wherein the positions corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain according to EU numbering are F, E, and A, respectively, in the first and second heavy chains. ### Claim 12 (depends on 11) The multispecific antibody or fragment thereof of claim 11, wherein (i) the amino acid in the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said first heavy chain, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said second heavy chain, or (ii) the amino acid in the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said first heavy chain, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said second heavy chain. ### Claim 13 (depends on 11) The multispecific antibody or fragment thereof according to claim 11, wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117. ### Claim 14 (depends on 11) The multispecific antibody or fragment thereof according to claim 11, wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123. ### Claim 15 (depends on 11) The multispecific antibody or fragment thereof according to claim 11, wherein the first VL region comprises the amino acid sequence as set forth in SEQ ID NO:121. ### Claim 16 (depends on 11) The multispecific antibody or fragment thereof according to claim 11, wherein the second VL region comprises the amino acid sequence as set forth in SEQ ID NO: 127. ### Claim 17 A multispecific antibody or a fragment thereof comprising a first heavy chain and a first light chain comprising a first antigen-binding region capable of binding to human CD40, and a second heavy chain and a second light chain comprising a second antigen-binding region capable of binding to human CD137, wherein the first antigen-binding region capable of binding to human CD40 comprises a first heavy chain variable (VH) region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 1, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 2, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 3, and a first light chain variable (VL) region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 4, a LCDR2 sequence comprising the amino acid sequence YTS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 5; wherein the second antigen-binding region capable of binding to human CD137 comprises a second VH region comprising an HCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 64, an HCDR2 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 65, and an HCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 66, and a second VL region comprising a LCDR1 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 67, a LCDR2 sequence comprising the amino acid sequence GAS, and a LCDR3 sequence comprising the amino acid sequence as set forth in SEQ ID NO: 68; wherein the first and second heavy chains each comprise an amino acid substitution at one or more positions corresponding to EU index positions T366, L368, K370, D399, F405, Y407, and/or K409 in a human IgG1 heavy chain, and wherein the first and second heavy chains are not substituted in the same positions, wherein in at least one of said first and second heavy chains one or more amino acids in the positions corresponding to positions L234, L235, D265, N297, and P331 in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively. ### Claim 18 (depends on 17) The multispecific antibody or fragment thereof according to claim 17, wherein the first VH region comprises the amino acid sequence as set forth in SEQ ID NO:117. ### Claim 19 (depends on 17) The multispecific antibody or fragment thereof according to claim 17, wherein the second VH region comprises the amino acid sequence as set forth in SEQ ID NO:123. ### Claim 20 (depends on 17) The multispecific antibody or fragment thereof according to claim 17, wherein the first VL region comprises the amino acid sequence as set forth in SEQ ID NO:121. ### Claim 21 (depends on 17) The multispecific antibody or fragment thereof according to claim 17, wherein the second VL region comprises the amino acid sequence as set forth in SEQ ID NO: 127.