# US 7,608,267 B2 · Multifunctional cytokines > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,608,267 B2 - Document ID: US7608267B2 - Status: Granted - Kind: B2 - Application number: 10/895,396 - Filed: 2004-07-21 - Published: 2005-03-24 - Granted: 2009-10-27 - Earliest claimed: 2003-07-21 - Assignees: Transgene S.A. - Inventors: Stéphane Paul - Web page: https://patentgrep.com/patent/US7608267B2 ## CPC - C12N 15/62: DNA sequences coding for fusion proteins - A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 31/20: for DNA viruses - A61P 33/06: Antimalarials - A61P 35/00: Antineoplastic agents - A61P 37/04: Immunostimulants - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C07K 14/54: Interleukins [IL] - C07K 14/5418: {IL-7} - C07K 14/5443: {IL-15} - C07K 14/55: IL-2 - C07K 14/57: IFN-gamma - 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) - C07K 2319/00: Fusion polypeptide - C12N 2799/022: where the vector is derived from an adenovirus - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) ## Abstract The present invention relates to a novel fusion protein with the formula X—Y, or Y—X, wherein X represents a first immunoregulating polypeptide and Y represents a second immunoregulating polypeptide different from X; it also relates to a nucleic acid molecule encoding such a fusion protein and a vector comprising such a nucleic acid molecule; it also relates to infectious viral particles and host cells comprising such a nucleic acid molecule or such a vector as well as a process for producing such infectious viral particles; is also relates to a method for recombinantly producing such a fusion protein; and, lastly, it also relates to a pharmaceutical composition comprising such a fusion protein, a nucleic acid molecule, a vector, infectious viral particles and a host cell as well as the therapeutic use thereof. ## Claims ### Claim 1 A fusion protein with the formula: X—Y wherein X represents IL-2; and Y represents IL-18 wherein said IL-18 is a pro-IL-18; and wherein said pro-IL-18 is fused to the carboxy terminus of said IL-2. ### Claim 2 (depends on 1) The fusion protein of claim 1, wherein said IL-18 is an IL-18 variant selected from among the group consisting of: a variant K89A having the lysine residue in position 89 of the corresponding native IL-18 substituted by an alanine residue. ### Claim 3 (depends on 1) The fusion protein of claim 1, wherein said fusion protein comprises the amino acid sequence of SEQ ID NO: 9. ### Claim 4 (depends on 1) A nucleic acid molecule encoding the fusion protein of claim 1. ### Claim 5 (depends on 4) A vector comprising the nucleic acid molecule of claim 4. ### Claim 6 (depends on 5) The vector of claim 5, wherein said vector is a bacterial plasmid, bacteriophage, yeast episome, artificial chromosome, or a viral vector selected from the group consisting of baculovirus vector, papovavirus vector, herpes virus vector, adenovirus vector, adenovirus-associated virus (AAV) vector, poxvirus vector, foamy virus vector, and retrovirus vector. ### Claim 7 (depends on 6) The vector of claim 6, wherein said vector is an adenoviral vector. ### Claim 8 (depends on 7) The vector of claim 7, wherein said vector is an E1- and E3-deleted replication-defective adenoviral vector comprising said nucleic acid molecule inserted in replacement of the E1 region and placed under the control of the CMV promoter. ### Claim 9 (depends on 5) The vector of claim 5, wherein said vector further comprises one or more transgenes encoding a polypeptide selected from the group consisting of tumor proliferation inhibitors and antigens against which an immune response is desired. ### Claim 10 (depends on 9) The vector of claim 9, wherein said tumor proliferation inhibitor is a polypeptide possessing both CDase (cytosine deaminase) and UPRTase (uracil phosphoribosyl transferase) activities of Saccharomyces cerevisiae. ### Claim 11 (depends on 9) The vector of claim 9, wherein said specific antigen is a human papilloma virus (HPV) antigen. ### Claim 12 (depends on 11) The vector of claim 11, wherein said HPV antigen is a membrane-anchored form of a non-oncogenic variant of the early HPV-16 E7 antigen. ### Claim 13 (depends on 4, 5) An isolated host cell comprising the nucleic acid molecule according to claim 4 or the vector according to claim 5. ### Claim 14 (depends on 5) A method for producing a fusion protein, comprising introducing a vector according to claim 5 into a suitable host cell to produce a transfected or infected host cell, culturing in-vitro said transfected or infected host cell under conditions suitable for growth of the host cell, and thereafter recovering said fusion protein from said culture, and optionally, purifying said recovered fusion protein. ### Claim 15 (depends on 1, 5, 13) A pharmaceutical composition comprising an effective amount of the fusion protein according to claim 1, the vector according to claim 5, the host cell according to claim 13 or any combination thereof and optionally a pharmaceutically acceptable vehicle. ### Claim 16 (depends on 1, 5, 13, 15) A method for enhancing an immune response in an animal or human organism comprising introducing into said organism the fusion protein according to claim 1, the vector according to claim 5, the host cell according to claim 13 or the composition of claim 15, so as to enhance said immune response. ### Claim 17 (depends on 1) The fusion protein of claim 1, wherein said IL-2 is a native IL-2. ### Claim 18 (depends on 1) The fusion protein of claim 1, wherein said pro-IL-18 is a native pro-IL-18. ### Claim 19 (depends on 1) The fusion protein of claim 1 wherein said pro-IL-18 is fused to the carboxy terminus of IL-2 by a peptide linker. ### Claim 20 (depends on 19) The fusion protein of claim 19, wherein the peptide linker consists of two copies of the sequence Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 60). ### Claim 21 (depends on 1) An adjuvant composition to enhance the immune response of an animal or human organism to an antigen, wherein the adjuvant composition comprises the fusion protein of claim 1 and the antigen. ### Claim 22 (depends on 3) An adjuvant composition to enhance the immune response of an animal or human organism to an antigen, wherein the adjuvant composition comprises the fusion protein of claim 3 and the antigen. ### Claim 23 (depends on 4) An adjuvant composition to enhance the immune response of an animal or human organism to an antigen, wherein the adjuvant composition comprises the nucleic acid molecule of claim 4 and a nucleic acid molecule encoding the antigen. ### Claim 24 (depends on 4, 23) The adjuvant composition of claim 23, wherein the nucleic acid molecule of claim 4 and time nucleic acid molecule encoding the antigen are comprised in the same vector. ### Claim 25 (depends on 4, 23) The adjuvant composition of claim 23, wherein the nucleic acid molecule of claim 4 and the nucleic acid molecule encoding the antigen are comprised in two different vectors.