# US 7,612,173 B2 · Vaccine for prevention and treatment of HIV-infection > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,612,173 B2 - Document ID: US7612173B2 - Status: Granted - Kind: B2 - Application number: 11/573,128 - Filed: 2005-08-03 - Published: 2007-10-18 - Granted: 2009-11-03 - Earliest claimed: 2004-08-05 - Assignees: Glaxosmithkline Biologicals SA - Inventors: Helge Abrecht; Martine Delchambre; Martine Marchand; Nathalie Louise Mathy; Philippe Jean Gervais Ghislain Permanne; Gerald Hermann Voss - Web page: https://patentgrep.com/patent/US7612173B2 ## CPC - C07K 14/005: from viruses - A61P 31/18: for HIV - A61P 37/04: Immunostimulants - A61K 2039/645: {Dendrimers; Multiple antigen peptides} - C07K 2319/00: Fusion polypeptide - C12N 2740/16222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2740/16322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract This invention relates to novel HIV polypeptide and polynucleotide fusions of Gag, Pol and Nef which are useful in immunogenic compositions and vaccines. The invention relates in particular to a polypeptide which comprises Nef or an immunogenic fragment thereof, and p17 Gag and/or p24 Gag or immunogenic fragments thereof, wherein when both p17 and p24 Gag are present there is at least one HIV antigen or immunogenic fragment between them. The polypeptide may also comprise Pol or RT or an immunogenic fragment thereof. ## Claims ### Claim 1 A fusion polypeptide comprising a Nef polypeptide, a reverse transcriptase (RT) polypeptide, a p17 Gag polypeptide, and a p24 Gag polypeptide, wherein there is at least one HIV antigen between the p17 Gag polypeptide and the p24 Gag polypeptide. ### Claim 2 (depends on 1) The fusion polypeptide of claim 1, wherein the RT polypeptide is p66. ### Claim 3 (depends on 1) The fusion polypeptide of claim 1, wherein the RT polypeptide is truncated at the C terminus such that it lacks the carboxy terminal RNase H domain. ### Claim 4 (depends on 1) The fusion polypeptide of claim 1, wherein the RT polypeptide is p51. ### Claim 5 (depends on 1) The fusion polypeptide of claim 1, wherein the RT polypeptide comprises a mutation at the amino acid position corresponding to position 592 in SEQ ID NO:2 where methionine is replaced by another amino acid residue. ### Claim 6 (depends on 1) The fusion polypeptide of claim 1, comprising from N-terminal to C-terminal: p24-RT-Nef-p17. ### Claim 7 (depends on 1) A process for purifying the fusion polypeptide of claim 1, the process comprising: i). providing a composition comprising the unpurified fusion polypeptide; ii). subjecting the composition to at least two chromatographic steps; iii). optionally carboxyamidating the fusion polypeptide; and iv) performing a buffer exchange step to provide the fusion polypeptide in a suitable buffer for a pharmaceutical formulation. ### Claim 8 (depends on 7) The process of claim 7, wherein there are no more than two chromatographic steps. ### Claim 9 (depends on 7) The process of claim 7, wherein the fusion polypeptide is carboxyamidated, and the carboxyamidation is performed between the at least two chromatographic steps. ### Claim 10 (depends on 5) The fusion polypeptide of claim 5, wherein methionine is replaced by lysine. ### Claim 11 (depends on 1) A pharmaceutical composition comprising the fusion polypeptide of claim 1. ### Claim 12 (depends on 11) The pharmaceutical composition of claim 11, further comprising a Th1 inducing adjuvant. ### Claim 13 (depends on 12) The pharmaceutical composition of claim 12, wherein the TH1 inducing adjuvant comprises QS21, 3D-MPL, or a combination of QS21 and 3D-MPL. ### Claim 14 (depends on 1) A pharmaceutical kit comprising: a) a pharmaceutical composition comprising the fusion polypeptide of claim 1; and b) a second pharmaceutical composition comprising a polynucleotide encoding at least one of a Nef epitope and a Gag epitope, wherein the at least one Nef epitope and Gag epitope are present in the polypeptide of a). ### Claim 15 (depends on 14) The pharmaceutical composition of claim 14, wherein the RT polypeptide is selected from the group consisting of: full length RT and RT truncated at the C terminus such that it lacks the carboxy terminal RNase H domain. ### Claim 16 (depends on 1) A pharmaceutical kit comprising: a) a pharmaceutical composition comprising a polynucleotide encoding the fusion polypeptide of claim 1; and b) a second pharmaceutical composition comprising a polypeptide comprising at least one of a Nef epitope and a Gag epitope, wherein the at least one Nef epitope and Gag epitope are present in the polypeptide of a). ### Claim 17 (depends on 16) The pharmaceutical composition of claim 16, wherein the RT polypeptide is selected from the group consisting of: full length RT and RT truncated at the C terminus such that it lacks the carboxy terminal RNase H domain. ### Claim 18 (depends on 1) The fusion polypeptide of claim 1, comprising from N-terminal to C-terminal: p24-RT-Nef-p17, wherein the amino acid at the position corresponding to position 592 in SEQ ID NO:2 is not methionine. ### Claim 19 (depends on 18) The fusion polypeptide of claim 18 where said amino acid is lysine. ### Claim 20 (depends on 1) The fusion polypeptide of claim 1, comprising from N-terminal to C-terminal: p24-p51RT-Nef-p17. ### Claim 21 (depends on 1) The fusion polypeptide of claim 1, comprising from N-terminal to C-terminal: p22-p51RT-Nef-p17, wherein the amino acid at the position corresponding to position 592 in SEQ ID NO:2 is not methionine. ### Claim 22 (depends on 21) The fusion polypeptide of claim 21 where said amino acid is lysine. ### Claim 23 (depends on 1) The fusion polypeptide of claim 1, comprising SEQ ID NO:2. ### Claim 24 (depends on 1) The fusion polypeptide of claim 1, comprising SEQ ID NO:15. ### Claim 25 (depends on 1) The fusion polypeptide of claim 1, comprising SEQ ID NO:19. ### Claim 26 (depends on 1) The fusion polypeptide of claim 1, where said RT polypeptide comprises a point mutation that removes reverse transcriptase enzyme activity, wherein said point mutation is the substitution of lysine for tryptophan at the amino acid position corresponding to position 464 in SEQ ID NO:2.