Composition containing nucleic acids, preparation and uses

Details Composition containing nucleic acids, preparation and uses Composition containing nucleic acids, preparation and uses

1996-07-10

1998-12-08

Low, Christopher S. F.

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Carbohydrate doai

514 18, 514 19, 435455, 435458, 435325, 4353201, 435 691, A01N 4304

Patent

active

058469478

DESCRIPTION:

BRIEF SUMMARY
This application is a national phase application under 35 U.S.C. 371 of PCT/FR95/00022, filed Jan. 9, 1995 with a claim to the priority of French application FR94/00159, filed Jan. 10, 1994.
The present invention relates to compositions based on nucleic acids, to their preparation and to their use. More particularly, it relates to compositions comprising at least one nucleic acid and one lipopolyamine and to their use in gene therapy, especially for the transfer of nucleic acids.
Gene therapy consists in correcting a deficiency or abnormality (mutation, aberrant expression, and the like) or in providing for the expression of a protein of therapeutic interest by introduction of genetic information into the affected cell or organ. This genetic information can be introduced either in vitro into a cell extracted from the organ, the modified cell then being reintroduced into the organism, or directly in vivo into the appropriate tissue. Various techniques have been described for the transfer of this genetic information, including various transfection techniques involving complexes of DNA and DEAE-dextran (Pagano et al., J. Virol., 1 (1967), 891), of DNA and nuclear proteins (Kaneda et al., Science, 243 (1989), 375), of DNA and lipids (Felgner et al., PNAS, 84 (1987), 7413) and of DNA and polylysine, the use of liposomes (Fraley et al., J. Biol. Chem., 255 (1980), 10431), and the like. More recently, the use of viruses as vectors for gene transfer has appeared as a promising alternative to these physicochemical transfection techniques. In this respect, various viruses have been tested for their ability to infect certain cell populations. In particular, retroviruses (RSV, HMS, MMS, and the like), the HSV virus, adeno-associated viruses and adenoviruses.
However, the techniques developed until now do not make it possible to satisfactorily resolve the difficulties related to the transfer of genes into cells and/or the organism. In particular, the problems related to the penetration of nucleic acid into cells are not completely solved. In fact, the polyanionic nature of nucleic acids prevents their passage through cell membranes. While it has been shown that naked nucleic acids are capable of passing through the plasma membrane of certain cell types in vivo (see especially Application No. WO 90/11092), transfection efficiency remains fairly low. Moreover, naked nucleic acids have a short plasma half-life, due to their degradation by enzymes and their removal by urinary routes. Moreover, while recombinant viruses make it possible to improve the efficiency of transfer of nucleic acids, their use presents certain risks such as pathogenicity, transmission, replication, recombination, transformation, immunogenicity, and the like.
The present invention introduces an advantageous solution to these various problems. The Applicant has in fact shown that certain compositions comprising a nucleic acid and a lipopolyamine can make possible the in vivo transfer of the said nucleic acid into a cell and/or organ with high efficiency and without toxicity. The compositions of the invention also make it possible to avoid the disadvantages related to the use of viral vectors (potential dangers, limited size of transferred gene, high cost, and the like).
The use of certain lipopolyamines for the in vitro transfection of cell cultures has already been described in the prior art. Thus, Application EP 394,111 describes the use of certain lipopolyamines for the in vitro transfection of cell lines. The article by Demeneix et al. (Int. J. Dev. Biol., 35 (1991), 481) likewise describes the use of a lipopolyamine (dioctadecylamidoglycylspermine, DOGS) for the in ovo transfection of nucleic acids. According to these documents, the lipopolyamines must be used under conditions such that the positive charges of the lipopolyamine
egative charges of the nucleic acid ratio is between 2 and 5 and preferably equal to 3 or 4. However, as shown in Examples 8 and 9 of the present application, none of the conditions described in these documents, surprisin

REFERENCES:
patent: 5171678 (1992-12-01), Behr et al.
patent: 5616745 (1997-04-01), Behr et al.
Felgner, P. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. PNAS 84:7413-7417, Nov. 1, 1987.
Gao, X. et al. A novel cationic liposome reagent for efficient transfection of mammalian cells.Biochem. Biophys. Res. Comm. 179:280-285, Aug. 30, 1991.
Yu, Ach, et al. Inhibition of GFAP synthesis by antisense RNA in astrocytes. J. Neuroscience Research 30:72-79, 1991.
Uhlmann, E. et al. Antisense oligonucleotides: a new therapeutic principle. Chemical Reviews 90:544-584, Jun. 1, 1990.
Nicolau, C, et al. In vivo expression of rat insulin after intravenous administration of the liposome-entrapped gene for rat insulin I. PNAS 80:1068-1072, Feb. 1, 1983.
Ledley (Human Gene Therapy: 6:1129-1144, 1995).
Zelphati et al. (J. Controlled Release, 1996, 41:99-119).
Gao et al. (J Liposome Research, 3, 1:17-20, 1993).
Behr, J-P, Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA, Proceeding of The National Academy of Science of USA, 86, 6982-6986 (1989).
Loeffler, J.P., Gene transfer into primary and established mammalian cell lines with lipopolyamine-coated DNA, Methods in Enzymology, 217, 599-618 (1993).
Gao, et al, Cationic liposomes and polymers for gene transfer, Journal of Liposome Research, 3, 17-30 (1993).
Demeneix, et al, Gene transfer into intact vertebrate embryos, The International Journal of Developmental Biology, 35, 481-484 (1991).
Stewart, et al, Gene transfer in vivo with DNA-liposome complexes: Safety and acute toxicity in mice, Human Gene Therapy, 3, 267-275 (1992).
Malone, et al, Cationic liposome-mediated RNA transfection, Proceedings of the National Academy of Sciences of USA, 86, 6077-6081 (1989).

Affiliated with

Also associated with

Rating

Composition containing nucleic acids, preparation and uses does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Composition containing nucleic acids, preparation and uses, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Composition containing nucleic acids, preparation and uses will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-177351