Process of making cationic lipid-nucleic acid complexes

{
  "publication_number": "US6271208B1",
  "country": "US",
  "kind": "B1",
  "title": "Process of making cationic lipid-nucleic acid complexes",
  "abstract": "The invention is related to a method for preparing an homogenous suspension of stable lipid-nucleic acid complexes or particles, comprising: a) combining one or more cationic lipids, one or more colipids, and one or more stabilizing additives to form a lipid suspension, b) combining the lipid suspension with a nucleic acid to form a complex or a particle, and optionally c) subjecting the complex or particle to a sizing procedure. It also concerns an homogenous suspension produced notably by the above method.",
  "claims": [
    "1. A method for preparing an homogenous suspension of stable lipid-nucleic acid complexes or particles, comprising: a) combining one or more cationic lipids, one or more colipids, and one or more stabilizing additives to form a lipid suspension, b) combining the lipid suspension with a nucleic acid to form a complex or a particle, and optionally c) subjecting the complex or particle to a sizing procedure.",
    "2. The method of claim 1, further comprising the step of subjecting the lipid suspension to a sizing procedure to form a lipid suspension of particles of homogenous size.",
    "3. The method of claim 1, wherein the sizing procedure comprises extruding the lipid-nucleic acid complexes or particles through a membrane of defined pore diameter.",
    "4. The method of claim 3, wherein the lipid-nucleic acid complexes or particles are extruded through membranes having pore sizes in the range of 50 to 500 nm.",
    "5. The method of claim 1, wherein the lipid-nucleic acid complexes or particles in the homogeneous suspension have a particle size of 500 nm or less.",
    "6. The method of claim 1, wherein the lipid-nucleic acid complexes or particles in the homogeneous suspension have a particle size of 200 nm or less.",
    "7. The method of claim 1, wherein the cationic lipid or lipids are selected from the group consisting of spermidine-cholesterol, spermine-cholesterol, 3β[N-(N′,N′-dimethylaminoethane)-carbamoyl] cholesterol, dioctadecylamidoglycylspermine, and mixtures thereof.",
    "8. The method of claim 1, wherein the colipid is dioleoylphosphatidylethanolamine.",
    "9. The method of claim 1, wherein the stabilizing additive is polyethylene glycol coupled to a colipid.",
    "10. The method of claim 1, wherein the stabilizing additive is selected from the group consisting of perfluorinated or partially fluorinated alkyl chains coupled to colipid.",
    "11. The method claim 1, wherein the stabilizing additive is polyglucuronic acid coupled to a colipid.",
    "12. The method of claim 1, wherein the stabilizing additive is distearoylphosphatidylethanolamine-polyethylene glycol.",
    "13. The method of claim 9, wherein the moiety is a phospholipid.",
    "14. The method of claim 13, wherein the moiety is a zwitterionnic phospholipid.",
    "15. The method of claim 1, wherein the nucleic acid is selected from the group consisting of genomic DNA, cDNA, synthetic DNA, RNA, mRNA, ribozymes, antisense RNA and oligonucleotides.",
    "16. The method of claim 15, wherein the nucleic acid comprises a plasmid.",
    "17. An homogenous suspension of stable lipid-nucleic acid complexes or particles, produced by: a) combining one or more cationic lipids, one or more colipids, and one or more stabilizing additives to form a lipid suspension, b) combining the lipid suspension with a nucleic acid to form a complex or a particle, and optionally c) subjecting the complex or particle to a sizing procedure.",
    "18. The suspension of claim 17, wherein the lipid suspension of step a) is subjected to a sizing procedure to form a lip suspension of particles of homogenous size.",
    "19. The suspension of claim 17, wherein the sizing procedure comprises extruding the lipid-nucleic acid complexes or particles through a membrane of defined pore diameter.",
    "20. The suspension of claim 17, wherein the cationic lipids include mixtures of cationic lipids.",
    "21. The suspension of claim 17, wherein the complexes or particles in the homnogeneous suspension have a particle size of 500 nm or less.",
    "22. The suspension of claim 17, wherein the complexes or particles in the homogeneous suspension have a particle size of 200 nm or less.",
    "23. A lipid-nucleic acid complex comprising one or more cationic lipids, one or more colipids, one or more stabilizing additives, and a nucleic acid component.",
    "24. The complex of claim 23, wherein the complex has a particle size of 500 nm or less.",
    "25. The complex of claim 23, wherein the complex has a particle size of 200 nm or less.",
    "26. A pharmaceutical composition comprising the suspension of claim 17 and a pharmaceutically acceptable carrier.",
    "27. A pharmaceutical composition comprising the complex of claim 23 and a pharmaceutically acceptable carrier.",
    "28. A method for the delivery of a nucleic acid to the cells of an individual in need of such nucleic acid, comprising administering the composition of claim 26 to the cells of such an individual.",
    "29. The method of claim 28, wherein the nucleic acid is selected from the group consisting of genomic DNA, cDNA, synthetic DNA, RNA, mRNA, ribozymes, antisense RNA and oligonucleotides.",
    "30. The method of claim 29, wherein the nucleic acid comprises a plasmid.",
    "31. The method of claim 2, wherein the sizing procedure comprises extruding the lipid suspension through a membrane of defined pore diameter.",
    "32. The method of claim 31, wherein the lipid suspension is extruded through membranes having pore sizes in the range of 50 to 500 nm.",
    "33. The suspension of claim 18, wherein the sizing procedure comprises extruding the lipid suspension through a membrane of defined pore diameter.",
    "34. The suspension of claim 19, wherein the lipid-nucleic acids complexes or particles are extruded through membranes having pore sizes in the range of 50 to 500 nm.",
    "35. The suspension of claim 33, wherein the lipid suspension is extruded through membranes having pore sizes in the range of 50 to 500 nm."
  ],
  "description_excerpt": "The present invention is directed to stable complexes of cationic lipids and nucleic acid that can be used to deliver nucleic acid to a cell for the purpose of providing a therapeutic molecule to the cells of an individual in need of such treatment, and to methods for the preparation of stable, cationic lipid-nucleic acid complexes.\n\nSuccessful gene therapy depends on the efficient delivery to and expression of genetic information within the cells of a living organism. Most delivery mechanisms used so far involve viral vectors. Viruses have developed diverse and highly sophisticated mechanisms to achieve this goal including crossing of the cellular membrane, escape from lysosomal degradation, delivery of their genome to the nucleus and, consequently, have been used in many gene delivery applications.\n\nNon-viral vectors, which are based on receptor-mediated mechanisms (Perales et al., Eur. J. Biochem. 226:255-266, 1994; Wagner et al., Advanced Drug Delivery Reviews 14:113-135, 1994) or on lipid-mediated transfection (Eelgner et al., Proc. Natl. Acad. Sci. U.S.A. 84:7413-7417 1987; Behr et al., Proc. Natl. Acad. Sci. U.S.A. 86:6982-6986, 1989; Gao et al., Biochem. Biophys. Res. Communic. 179:280-285, 1991; Behr, Bioconjugate Chemistry 5:382-389, 1994; Fahrhood et al., Annals New York Academy of Sciences, 716:23-35, 1994; Ledley, Human Gene Therapy 6:1129-1144, 1995) promise to have advantages with respect to large - scale production, reduced risks related to viral vectors, targeting of transfectable cells, lower immunogenicity and the capacity to deliver larger fragments of DNA.",
  "cpc": [
    "A61K 9/1277",
    "A61K 48/00",
    "A61K 9/1272"
  ],
  "ipc": [
    "A61K 48/00",
    "A61K 9/127"
  ],
  "assignees": [
    "Transgene SA"
  ],
  "inventors": [
    "Rainer Bischoff"
  ],
  "filing_date": "1997-08-25",
  "publication_date": "2001-08-07",
  "grant_date": "2001-08-07",
  "priority_date": "1996-08-26",
  "application_number": "US-25414999-A",
  "family_id": "8225285",
  "cited_by_count": 80,
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}