Compositions for the delivery of polynucleotides to cells

• US 6,656,734 B1
• Filed: 06/30/1998
• Issued: 12/02/2003
• Est. Priority Date: 07/01/1997
• Status: Expired due to Fees

• Current Assignee: Transgene S.A. (Compagnie Merieux Alliance SAS)
• Original Assignee: Transgene S.A. (Compagnie Merieux Alliance SAS)
• Inventors: Bischoff, Rainer, Kolbe, Hanno, Schughart, Klaus
• Primary Examiner(s): Nguyen, Dave T.
• Assistant Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US09/107,271
• Time in Patent Office: 1,981 Days
• Field of Search: 424/486, 556/137, 514/44, 435/455, 435/458, 435/461, 536/23.1, 536/23.2, 536/23.5, 536/24.1
• US Class Current: 435/455
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The present invention relates to a composition which is intended for transferring at least one therapeutically active substance into the interior of a target cell, characterized in that it comprises a mixture of at least one therapeutically active substance and at least one polar compound which is selected from a specific group of aprotic polar compounds. The therapeutically active substance is preferably a polynucleotide, and the polar compound is DPSO, which improves the ability of the polynucleotide to be transfected into the interior of the cells. The compositions according to the invention can be used as a diagnostic therapeutic, prophylactic or vaccinal medicament for treating the human or animal body by means of gene therapy.

Claims

1. A process for transferring a therapeutically active polynucleotide into a target cell in vitro, wherein said cell is brought into contact with a composition comprising a mixture of at least one therapeutically active polynucleotide and at least one aprotic polar compound selected from the group consisting of:

(a) formula I;

 

wherein R₁ and R₂ are identical or different alkyl radicals of 1 to 8 carbon atoms which are optionally substituted,

with the proviso that when one of R₁ or R₂ is a radical of 1 or 2 carbon atoms, the other is a radical of at least 3 carbon atoms;

wherein the volume of the aprotic polar compound represents from 5 to 50% of the total volume of the composition,

and wherein the polynucleotide prior to being part of the mixture, is free of any compound which facilitates its introduction into a target cell.

2. The process of claim 1, wherein said target cell is a mammalian cell.

3. The process of claim 1, wherein said target cell is an airways cell.

4. A process according to claim 1, further comprising the step of adjusting the temperature of the composition to approximately the same temperature as the cell before bringing it into contact with the cell.

5. The process according to claim 1, wherein said aprotic polar compound is selected from the group consisting of di-n-propyl sulphoxide (DPSO), and tetramethylene sulfoxide (TEMSO).

6. The process according to claim 1, wherein said polynucleotide is an antisense polynucleotide.

7. The process according to claim 1, wherein said polynucleotide is a ribozyme.

8. The process according to claim 1, wherein said polynucleotide comprises a gene of interest and elements which enable the said gene of interest to be expressed.

9. The process according to claim 8, wherein said polynucleotide encodes all or part of a polypeptide.

10. The process according to claim 9, wherein said polypeptide is an enzyme, a hormone, a cytokine, a membrane receptor, an antibody, a factor which regulates transcription, translation or replication, a polypeptide, a polypeptide which forms a membrane channel, a transport polypeptide, an adhesion molecule or a ligand.

11. The process according to claim 1, wherein said aprotic polar compound is in aqueous solution.

12. The process according to claim 1, wherein the volume of the polar compound represents from 15 to 20% of the total volume of the composition.

13. The process according to claim 9, wherein the said polypeptide exhibits an immunogenic activity.

14. The process according to claim 1, wherein R₁ and R₂ are linked in order to cyclize the molecule.

15. The process according to claim 1, wherein said polynucleotide encodes all or part of a polypeptide.

16. The process according to claim 9, wherein the polynucleotide encodes a CFTR protein, dystrophin, factor VII or factor IX, HPV E6/E7, MUC1, BRAC1, interferon, an interleukin, tumor necrosis factor alpha, or GM-CSF.

17. The process according to claim 9, wherein the polynucleotide is a tk gene of herpes simplex virus type 1, a retinoblastoma gene or a gene for p53.

18. The process according to claim 9, wherein the polynucleotide encodes F(ab)₂, Fab¹, Fab fragments, or an anti-idiotype immunoglobulin.