# US 5,981,505 A · Compositions and methods for delivery of genetic material > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,981,505 A - Document ID: US5981505A - Status: Granted - Kind: A - Application number: 08/979,385 - Filed: 1997-11-26 - Granted: 1999-11-09 - Earliest claimed: 1993-01-26 - Assignees: The Trustees of the University of Pennsylvania; The Wistar Institute - Inventors: David B. Weiner; William V. Williams; Bin Wang - Web page: https://patentgrep.com/patent/US5981505A ## CPC - C07K 14/005: from viruses - A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) - A61K 39/0011: {Cancer antigens} - A61K 39/001106: {Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ErbB4} - A61K 39/001151: {p53} - A61K 39/001152: {Transcription factors, e.g. SOX or c-MYC} - A61K 39/001164: {GTPases, e.g. Ras or Rho} - A61K 39/001196: {Fusion proteins originating from gene translocation in cancer cells} - A61K 39/12: Viral antigens - A61K 39/21: Retroviridae, e.g. equine infectious anemia virus - C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation - A61K 2039/53: {DNA (RNA) vaccination} - A61K 2039/545: {characterised by the dose, timing or administration schedule} - A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59} - A61K 2039/57: {characterised by the type of response, e.g. Th1, Th2} - C12N 2740/16034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2740/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2740/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2740/16234: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2740/16322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - Y02A 90/10: Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract Methods of inducing genetic material into cells of an individual and compositions and kits for practicing the same are disclosed. The methods comprise the steps of contacting cells of an individual with a polynucleotide function enhancer and administering to the cells, a nucleic acid molecule that is free of retroviral particles. The nucleic acid molecule comprises a nucleotide sequence that encodes a protein that comprises at least one epitope that is identical or substantially similar to an epitope of a pathogen antigen or an antigen associated with a hyperproliferative or autoimmune disease, a protein otherwise missing from the individual due to a missing, non-functional or partially functioning gene, or a protein that produces a therapeutic effect on an individual. Methods of prophylactically and therapeutically immunizing an individual against HIV are disclosed. Pharmaceutical compositions and kits for practicing methods of the present invention are disclosed. ## Claims ### Claim 1 A pharmaceutical composition comprising: a) a polynucleotide function enhancer; and b) a DNA molecule that comprises a DNA sequence that encodes an antigen; wherein said polynucleotide function enhancer is selected from the group consisting of bupivacaine and tetracaine and said DNA sequence operatively linked to regulatory sequences which control the expression of said DNA sequence. ### Claim 2 (depends on 1) The pharmaceutical composition of claim 1 wherein said composition comprises bupivacaine. ### Claim 3 (depends on 1) The pharmaceutical composition of claim 1 wherein said composition comprises tetracaine. ### Claim 4 (depends on 1) The pharmaceutical composition of claim 1 wherein said DNA molecule is a plasmid. ### Claim 5 (depends on 1) The pharmaceutical composition of claim 1 wherein said DNA sequence encodes a variable region of a T cell receptor. ### Claim 6 (depends on 1) The pharmaceutical composition of claim 1 wherein said DNA sequence encodes a pathogen antigen. ### Claim 7 (depends on 6) The pharmaceutical composition of claim 6 wherein said DNA sequence encodes an antigen from an intracellular pathogen. ### Claim 8 (depends on 7) The pharmaceutical composition of claim 7 wherein said antigen is a viral antigen. ### Claim 9 (depends on 7) The pharmaceutical composition of claim 7 wherein said pathogen is a virus selected from the group consisting of: human immunodeficiency virus, HIV; human T cell leukemia virus, HTLV; influenza virus; hepatitis A virus; hepatitis B virus; hepatitis C virus; human papilloma virus, HPV; Herpes simplex 1 virus, HSV1; Herpes simplex 2 virus, HSV2; Cytomegalovirus, CMV; Epstein-Barr virus, EBR; rhinovirus; and, coronavirus. ### Claim 10 (depends on 9) The pharmaceutical composition of claim 9 wherein said pathogen is Herpes simplex 2 virus, HSV2. ### Claim 11 (depends on 9) The pharmaceutical composition of claim 9 wherein said pathogen is Hepatitis B virus, HBV. ### Claim 12 (depends on 9) The pharmaceutical composition of claim 9 wherein said pathogen is human T cell leukemia virus, HTLV. ### Claim 13 (depends on 9) The pharmaceutical composition of claim 9 wherein said pathogen is a human immunodeficiency virus. ### Claim 14 (depends on 1) The pharmaceutical composition of claim 1 wherein said DNA sequence encodes a hyperproliferative disease associated protein. ### Claim 15 (depends on 14) The pharmaceutical composition of claim 14 wherein said hyperproliferative disease is cancer. ### Claim 16 (depends on 14) The pharmaceutical composition of claim 14 wherein said hyperproliferative disease is a lymphoma. ### Claim 17 (depends on 14) The pharmaceutical composition of claim 14 wherein said hyperproliferative disease is a melanoma. ### Claim 18 A method of immunizing an individual comprising the steps of: injecting into tissue of said individual at a site on said individual's body, a DNA molecule and a polynucleotide function enhancer, said DNA molecule comprising a DNA sequence that encodes an antigen, said DNA sequence operatively linked to regulatory sequences which control the expression of said DNA sequence, said polynucleotide function enhancer selected from the group consisting of bupivacaine and tetracaine; wherein said DNA molecule is taken up by cells in said tissue, said DNA sequence is expressed in said cells and an immune response is generated against said antigen. ### Claim 19 (depends on 18) The method of claim 18 wherein said tissue includes skin and skeletal muscle. ### Claim 20 (depends on 18) The method of claim 18 wherein said tissue is skin. ### Claim 21 (depends on 18) The method of claim 18 wherein said tissue is muscle. ### Claim 22 (depends on 18) The method of claim 18 wherein said tissue is skeletal muscle. ### Claim 23 (depends on 18) The method of claim 18 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 24 (depends on 18) The method of claim 18 wherein said polynucleotide function enhancer is tetracaine. ### Claim 25 (depends on 18) The method of claim 18 wherein said DNA molecule is a plasmid. ### Claim 26 (depends on 18) The method of claim 18 wherein said immune response generated against said antigen is an immune response against a pathogen antigen. ### Claim 27 (depends on 26) The method of claim 26 wherein said pathogen is an intracellular pathogen. ### Claim 28 (depends on 27) The method of claim 27 wherein said intracellular pathogen is a virus. ### Claim 29 (depends on 26) The method of claim 26 wherein said pathogen is a virus selected from the group consisting of: human immunodeficiency virus, HIV; human T cell leukemia virus, HTLV; influenza virus; hepatitis a virus; hepatitis B virus; hepatitis C virus; human papilloma virus, HPV; Herpes simplex 1 virus, HSV1; Herpes simplex 2 virus, HSV2; Cytomegalovirus, CMV; Epstein-Barr virus, EBR; rhinovirus; and, coronavirus. ### Claim 30 (depends on 18) The method of claim 18 wherein said immune response generated against said antigen provides a protective immune response against a pathogen and said individual is immunized against said pathogen. ### Claim 31 (depends on 30) The method of claim 30 wherein said tissue is skin. ### Claim 32 (depends on 30) The method of claim 30 wherein said tissue is skeletal muscle. ### Claim 33 (depends on 30) The method of claim 30 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 34 (depends on 30) The method of claim 30 wherein said polynucleotide function enhancer is tetracaine. ### Claim 35 (depends on 30) The method of claim 30 wherein said DNA molecule is a plasmid. ### Claim 36 (depends on 30) The method of claim 30 wherein said antigen is a pathogen antigen. ### Claim 37 (depends on 30) The method of claim 30 wherein said pathogen is an intracellular pathogen. ### Claim 38 (depends on 37) The method of claim 37 wherein said intracellular pathogen is a virus. ### Claim 39 (depends on 38) The method of claim 38 wherein said virus is selected from the group consisting of: human immunodeficiency virus, HIV; human T cell leukemia virus, HTLV; influenza virus; hepatitis A virus; hepatitis B virus; hepatitis C virus; human papilloma virus, HPV; Herpes simplex 1 virus, HSV1; Herpes simplex 2 virus, HSV2; Cytomegalovirus, CMV; Epstein-Barr virus, EBR; rhinovirus; and, coronavirus. ### Claim 40 (depends on 39) The method of claim 39 wherein said virus is Herpes simplex 2 virus, HSV2. ### Claim 41 (depends on 39) The method of claim 39 wherein said virus is Hepatitis B virus, HBV. ### Claim 42 (depends on 39) The method of claim 39 wherein said virus is human T cell leukemia virus, HTLV. ### Claim 43 (depends on 18) The method of claim 18 wherein said immune response generated against said antigen provides a therapeutic immune response against a pathogen in an individual who is infected with said pathogen. ### Claim 44 (depends on 43) The method of claim 43 wherein said tissue is skin. ### Claim 45 (depends on 43) The method of claim 43 wherein said tissue is skeletal muscle. ### Claim 46 (depends on 43) The method of claim 43 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 47 (depends on 43) The method of claim 43 wherein said polynucleotide function enhancer is tetracaine. ### Claim 48 (depends on 43) The method of claim 43 wherein said DNA molecule is a plasmid. ### Claim 49 (depends on 43) The method of claim 43 wherein said pathogen is an intracellular pathogen. ### Claim 50 (depends on 43) The method of claim 43 wherein said pathogen is a virus. ### Claim 51 (depends on 18) The method of claim 18 wherein said immune response generated against said antigen is an immune response against a hyperproliferative disease-associated protein. ### Claim 52 (depends on 51) The method of claim 51 wherein said tissue is skin. ### Claim 53 (depends on 51) The method of claim 51 wherein said tissue is skeletal muscle. ### Claim 54 (depends on 51) The method of claim 51 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 55 (depends on 51) The method of claim 51 wherein said polynucleotide function enhancer is tetracaine. ### Claim 56 (depends on 51) The method of claim 51 wherein said DNA molecule is a plasmid. ### Claim 57 (depends on 51) The method of claim 51 wherein said hyperproliferative disease-associated protein is selected from the group consisting of: protein products of oncogenes myb, myc, fyn, ras, src, neu and trk; protein products of translocation gene bcr/abl; P53; variable regions of antibodies made by B cell lymphomas; and variable regions of T cell receptors of T cell lymphomas. ### Claim 58 (depends on 51) The method of claim 51 wherein said immune response generated against said antigen is a therapeutically effective immune response against a hyperproliferative disease-associated protein in an individual who has a hyperproliferative disease. ### Claim 59 (depends on 58) The method of claim 58 wherein said hyperproliferative disease is cancer. ### Claim 60 (depends on 58) The method of claim 58 wherein said hyperproliferative disease is a melanoma. ### Claim 61 (depends on 58) The method of claim 58 wherein said hyperproliferative disease is a lymphoma. ### Claim 62 (depends on 18) The method of claim 18 wherein said immune response generated against said antigen is an immune response against an autoimmune disease-associated protein. ### Claim 63 (depends on 62) The method of claim 62 wherein said tissue is skin. ### Claim 64 (depends on 62) The method of claim 62 wherein said tissue is skeletal muscle. ### Claim 65 (depends on 62) The method of claim 62 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 66 (depends on 62) The method of claim 62 wherein said polynucleotide function enhancer is tetracaine. ### Claim 67 (depends on 62) The method of claim 62 wherein said DNA molecule is a plasmid. ### Claim 68 (depends on 62) The method of claim 62 wherein said autoimmune disease associated-protein is selected from the group consisting of: variable regions of antibodies involved in B cell mediated autoimmune disease; and variable regions of T cell receptors involved in T cell mediated autoimmune disease. ### Claim 69 A method of introducing DNA molecules into cells of an individual comprising the step of: injecting into tissue of said individual at a site on said individual's body, DNA molecules and a polynucleotide function enhancer selected from the group consisting of bupivacaine and tetracaine, wherein said DNA molecules are taken up by cells in said tissue. ### Claim 70 (depends on 69) The method of claim 69 wherein said DNA molecule comprises a DNA sequence that encodes an protein, said DNA sequence operatively linked to regulatory sequences which control the expression of said DNA sequence. ### Claim 71 (depends on 69) The method of claim 69 wherein said tissue is skin. ### Claim 72 (depends on 69) The method of claim 69 wherein said tissue is skeletal muscle. ### Claim 73 (depends on 69) The method of claim 69 wherein said polynucleotide function enhancer is bupivacaine. ### Claim 74 (depends on 69) The method of claim 69 wherein said polynucleotide function enhancer is tetracaine. ### Claim 75 (depends on 69) The method of claim 69 wherein said DNA molecule is a plasmid.