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1. (WO1998050565) ENHANCED ADENOVIRUS-ASSISTED TRANSFECTION COMPOSITION AND METHOD
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/1998/050565 International Application No.: PCT/US1998/008852
Publication Date: 12.11.1998 International Filing Date: 01.05.1998
Chapter 2 Demand Filed: 23.11.1998
IPC:
A61K 48/00 (2006.01) ,C12N 15/861 (2006.01) ,C12N 15/87 (2006.01)
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
K
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
48
Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79
Vectors or expression systems specially adapted for eukaryotic hosts
85
for animal cells
86
Viral vectors
861
Adenoviral vectors
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
87
Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
Applicants:
THE REGENTS OF THE UNIVERSITY OF CALIFORNIA [US/US]; 5th floor 1111 Franklin Street Oakland, CA 94607-5200, US
Inventors:
GERMAN, Michael, S.; US
SZOKA, Francis, C., Jr.; US
Agent:
BERLINER, Robert; Fulbright & Jaworski L.L.P. 29th floor 865 S. Figueroa Street Los Angeles, CA 90017-2571, US
Priority Data:
08/852,93408.05.1997US
Title (EN) ENHANCED ADENOVIRUS-ASSISTED TRANSFECTION COMPOSITION AND METHOD
(FR) COMPOSITION ET PROCEDE DE TRANSFECTION ASSISTEE PAR ADENOVIRUS RENFORCEE
Abstract:
(EN) A composition for transfecting eukaryotic cells comprising a cationic polymer which has protonatable groups which serve to buffer the acidic endosome, protecting the endocytosed polynucleotide from degradation and a viral agent is used to target uptake into and/or lysis from endosomes in the desired eukaryotic cell. By co-infecting the eukaryotic cells with cationic polymer, polynucleotide, and the viral agent, the polynucleotide is brought into the cell and then released. Preferably, polyamidoamine dendrimers are used as the cationic polymer and adenovirus is used as the viral agent. The dendrimers help associate plasmid DNA with the adenov irus, which then provokes receptor-mediated endocytosis. Within the endosome, the tertiary amine groups of the dendrimer buffer the pH change in the endosome. Then, the endosomalytic activity of the adenovirus releases the plasmid DNA into the cell. Also preferably, the adenovirus and the dendrimers are mixed before addition to the polynucleotide and the cells are washed after about 1.5 hours of incubation.
(FR) Une composition de transfection de cellules eucaryotes contenant un polymère cationique comprenant des groupes protonables servant à tamponner l'endosome acide, empêchant la dégradation des polynucléotides endocytosées, et un agent viral est utilisé pour cibler l'apport et/ou la lyse à partir d'endosomes dans la cellule eucaryote souhaitée. La co-infection des cellules eucaryotes par le polymère cationique, le polynucléotide et l'agent viral permet d'amener et de libérer le polynucléotide dans la cellule. De préférence, des dendrimères de polyamidoamine sont utilisés comme polymère cationique et l'adénovirus est utilisé comme agent viral. Les dendrimères aident à associer l'ADN plasmidique à l'adénovirus, qui provoque alors une endocytose induite par récepteur. Dans l'endosome, les groupes d'amine tertiaire du dendrimère limitent la modification du pH dans l'endosome. Ensuite, l'activité endosomale de l'adénovirus libère l'ADN plasmidique dans la cellule. L'adénovirus et les dendrimères sont mélangés, de préférence, avant d'être ajoutés au polynucléotide et les cellules sont lavées au bout d'environ de 1,5 heure d'incubation.
Designated States: AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, GM, GW, HU, ID, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, UZ, VN, YU, ZW
African Regional Intellectual Property Organization (ARIPO) (GH, GM, KE, LS, MW, SD, SZ, UG, ZW)
Eurasian Patent Organization (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM)
European Patent Office (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)
Also published as:
AU1998072750