# US 10,308,696 B2 · IL2 and TNF immunoconjugates > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,308,696 B2 - Document ID: US10308696B2 - Status: Granted - Kind: B2 - Application number: 15/564,694 - Filed: 2016-05-05 - Published: 2018-03-22 - Granted: 2019-06-04 - Earliest claimed: 2015-05-08 - Assignees: Philogen S.P.A. - Applicants: Philogen S.P.A. - Inventors: Roberto De Luca; Francesca Pretto; Sarah Wulhfard - Web page: https://patentgrep.com/patent/US10308696B2 ## CPC - C07K 14/525: Tumour necrosis factor [TNF] - A61K 47/6813: {the drug being a peptidic cytokine, e.g. an interleukin or interferon} - A61K 47/6851: {the antibody targeting a determinant of a tumour cell} - A61P 35/00: Antineoplastic agents - C07K 14/55: IL-2 - C07K 16/18: against material from animals or humans - 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} - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - C07K 2319/74: containing a fusion for binding to a cell surface receptor - C07K 2319/75: containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones - 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 The present application relates to conjugates comprising interleukin 2 (IL2), and a tumour necrosis factor, such as tumour necrosis factor alpha (TNFα), and an antibody molecule. The antibody molecule preferably binds to an antigen associated with neoplastic growth and/or angiogenesis, such as the Extra-Domain A (EDA) of fibronectin. The conjugate may be used in the treatment of cancer. ## Claims ### Claim 1 A conjugate comprising interleukin-2 (IL2), tumor necrosis factor alpha (TNFα), and an antibody molecule which binds the Extra Domain-A (ED-A) of fibronectin. ### Claim 2 (depends on 1) The conjugate according to claim 1, wherein the antibody molecule is, or comprises, a single chain Fv (scFv). ### Claim 3 A conjugate comprising interleukin-2 (IL2), tumor necrosis factor alpha (TNFα), and an antibody molecule that binds the Extra Domain-A (ED-A) of fibronectin, wherein the antibody molecule comprises an antigen binding site having the complementarity determining regions (CDRs) of antibody F8 set forth in SEQ ID NOs 6-11. ### Claim 4 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule comprises the VH and VL domains of antibody F8 set forth in SEQ ID NOs 2 and 4. ### Claim 5 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule is an scFv, and wherein the VH domain and the VL domain of the scFv are linked by a 14 to 20 amino acid linker. ### Claim 6 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule has, or comprises, the amino acid sequence of scFv F8 set forth in SEQ ID NO: 5. ### Claim 7 (depends on 3) The conjugate according to claim 3, wherein the IL2 is human IL2. ### Claim 8 (depends on 7) The conjugate according to claim 7, wherein the IL2 comprises, or consists of, the sequence set forth in SEQ ID NO: 12. ### Claim 9 (depends on 3) The conjugate according to claim 3, wherein the TNFα is human TNFα. ### Claim 10 (depends on 9) The conjugate according to claim 9, wherein the TNFα comprises, or consists of, the sequence set forth in SEQ ID NO: 15, or the sequence set forth in SEQ ID NO: 40. ### Claim 11 (depends on 3) The conjugate according to claim 3, wherein the IL2 is linked to the antibody molecule by a peptide linker. ### Claim 12 (depends on 3) The conjugate according to claim 3, wherein the TNFα is linked to the antibody molecule via a peptide linker. ### Claim 13 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule is, or comprises, a single chain Fv (scFv) and, wherein the IL2 is linked to the N-terminus of the VH domain of the scFv via a peptide linker and the TNFα is linked to the C-terminus of the VL domain of the scFv via a peptide linker. ### Claim 14 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule is, or comprises, a single chain Fv (scFv) and, wherein the TNFα is linked to the N-terminus of the VH domain of the scFv via a peptide linker and the IL2 is linked to the C-terminus of the VL domain of the scFv via a peptide linker. ### Claim 15 (depends on 3) The conjugate according to claim 3, wherein the antibody molecule is, or comprises, a single chain Fv (scFv) and, wherein the IL2 and the TNFα are linked to C-terminus of the VL domain of the scFv via a peptide linker. ### Claim 16 (depends on 3) The conjugate according to claim 3, wherein the peptide linker is 10 to 20 amino acids long. ### Claim 17 (depends on 3) The conjugate according to claim 3, wherein the conjugate has, or comprises, the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 39. ### Claim 18 (depends on 3) The conjugate according to claim 3, wherein the conjugate has, or comprises, the amino acid sequence set forth in SEQ ID NO: 41 or SEQ ID NO: 42. ### Claim 19 (depends on 3) The conjugate according to claim 3, wherein the conjugate has, or comprises, the amino acid sequence set forth in SEQ ID NO: 43 or SEQ ID NO: 44. ### Claim 20 A conjugate comprising the amino acid sequence of SEQ ID NO: 1.