Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-06-30
1999-06-22
Ketter, James
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
4351723, 4353201, 536 231, C12Q 168, C12N 1563, C07H 2104
Patent
active
059142316
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the field of synthetic transfection systems useful in the delivery of gene constructs or DNA fragments to cells, especially to cells in living organisms. More in particular, the invention relates to cationic polymers having broad possibilities to be modified or adapted in order to create a flexible DNA gene delivery system, which can be used in, e.g., gene therapy applications.
Gene therapy is seen as a promising method to correct hereditary defects or to treat life threatening diseases such as cancer and AIDS. In gene therapy, nucleic acid fragments or gene constructs are brought into target cells. These nucleic acid fragments or gene constructs are preferably incorporated in plasmids or other vectors.
If the reconstructed plasmids are applied to an organism per se, this generally leads to low expression of the introduced gene, if any. There are three main reasons for this low expression. First, the plasmids will hardly ever reach the cell population where they are intended to be incorporated, due to degradation and elimination processes. Second, if the plasmids do reach the target cells, they cannot simply pass the cellular membrane, because of the strongly polar nature and the size of the plasmids. Third, if a plasmid does invade a target cell, it normally will be enclosed in an endosome, which will convert into a lysosome. In the lysosome, the plasmid will be degraded so that the incorporated gene cannot be expressed.
For the above reasons, in gene therapy plasmids comprising a desired gene construct are transported to and delivered in the target cells by means of carrier systems.
In recent years, many efforts have been made in the research on potentially suitable transfection systems, both of viral and non-viral origin. These transfection systems should deliver the desired gene to the target cell and cause it to be expressed to a high degree.
Viral vectors are very suitable, because by nature adapted, to introduce plasmids in target cells and to avoid endosome disruption, the degradation of the plasmids in endosomes or the transition of endosomes into lysosomes. However, viral vectors have a number of pronounced disadvantages. Viral vectors are able to effect integration of the introduced gene in the chromosomal DNA of the target cell. The site where this integration is effected cannot (yet) be predicted or controlled, which involves the risk of destroying essential genes or activation of, e.g., oncogenes. In addition, it is at present very difficult to provide for viral vectors on a commercial scale. Moreover, viral vectors generally trigger the immune system of a living organism, which will lead to immuno responses against the transfection system when used in vivo. Finally, viral vectors inherently set limits to the size of the gene construct to be introduced in the target cell.
In order to overcome the intrinsic disadvantages of viral vectors, synthetic transfection systems should offer good perspectives.
In this light, a review article of Kabanov et al. in Bioconjugate Chemistry vol.6, no. 1 (1995), 7-20 is mentioned. This review article describes in general terms the principle of delivery of genetic material onto cells employing soluble in terpolyelectrolyte complexes (IPEC's) of nucleic acids with linear polycations. It is indicated that various polycations have been used to produce IPEC's. As explicit examples polyvinyl pyrimidinium salts, polypeptides such as polylysine conjugates and lipopolylysines, and spermines are mentioned.
Explicit reference is made to research carried out by the group of E. Wagner, relating to gene delivery by means of plasmid-polylysine complexes (Curiel et al., Adenovirus Enhancement of Transferrin-Polylysine-Mediated Gene Delivery, Proc. Natl. Acad. Sci. 88 (1991) 8850-8854; Plank et al., Gene Transfer into Hepatocytes Using Asialloglycoprotein Receptor Mediated Endocytosis of DNA Complexed with an Artificial Tetra-Antennary Galactose Ligand Bioconj. Chem. 3 (1992) 533-539; Wagner et al., Influenza Virus Hemagglutin HA2 N-Terminal Fu
REFERENCES:
Kabanove et al., "DNA Complexes with Polycations for the Delivery of genetic Material into Cells," Bioconjugate Chemistry, vol. 6, No. 1 (1995), pp. 7-20.
Curiel et al., "Adenovirus Enhancement of Transferrin-Polysine-Medicated Gene Delivery," Proc. Natl. Acad. Sci., vol. 88 (1991) 8850-8854.
Plank et al., "Gene Transfer into Hepatocytes Using Asialloglycoprotein Receptor Mediated Endocytosis of DNA Complesed with an Artificial Tetra-Antennary Galactose Ligand," Bioconj. Chem., vol. 3, (1992) pp. 533-539.
Wagner et al., "Influenza Virus Hamagglutin HA2 N-Terminal Fusogenic Peptides Augment Gene Transfer By Transferrin-Polysine-DNA Complexes: Toward a synthetic virus-like gene-transfer vehicle," Proc. Natl. Acad. Sci., vol. 89 (1992) pp. 7934-7938.
Curiel et al., "Gene Transfer to Respiratory Epithelial Cells via the Receptor Mediated Endocytosis Pathway," Am. J. Respir. Cell. Mol. Biol., vol. 6 (1992) pp. 247-252.
Ph.D. Tesis of Jaap Goedemoed, "Polyphosphazene Drug Delivery Systems for Antitumor Treatment," University of Leiden (1990).
Crommen et al., "Poly(Organo) Phosphazenes," Biomaterials, vol. 13, (1992) pp. 511-520.
Domb et al., "Degradable Polymers for Site-Specific Drug Delivery," Polymer Advanced Technology, vol. 3, No. 6, (1992) pp. 279-292.
Calicietti et al., "Amino Acid esters and imidazole derivatives of ployposphazenes: Influence on release of drugs, Degradability and Swelling," Il Farmaco, vol. 49, No. 1, (1994) 69-74.
Andrianov et al., "Controlled release using ionotropic polyphosphazene hydrogels," Controlled Release, vol. 27, No. 1 (1983) pp. 69-77.
Victor A, McKusick, M.D., Mendelian Inheritance in man: Catalogs of autosomal dominant, autosomal recessive, and X-linked phenotypes, 8.sup.th edition. John Hopkins University Press (1988).
Plank et al., "The Influence of Endosome-Disruptive Peptides on Gene Transfer Using Synthetic VirusLike Gene Transfer Systems," J. Biol. Chem., vol. 269 (1994) pp. 12918-12924.
Mujumdar et al., "Solution Polymerization of Selected Polyphosphazenes," Makromol. Chem., vol. 190 (1989) pp. 2293-2302.
Bout Abraham
Hennink Wilhelmus Everhardus
Ketter James
Octoplus B.V.
Sandals William
Universiteit Utrecht Introgene B.V.
LandOfFree
Poly(organo)phosphazenes for use in synthetic transfection syste does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Poly(organo)phosphazenes for use in synthetic transfection syste, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Poly(organo)phosphazenes for use in synthetic transfection syste will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1707729