TRIBLOCK COPOLYMERS FOR CYTOPLASMIC DELIVERY OF GENE-BASED DRUGS

• US 20100222407A1
• Filed: 05/31/2006
• Published: 09/02/2010
• Est. Priority Date: 05/31/2005
• Status: Active Grant

• Current Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Inventors: Hubbell, Jeffrey A., Segura, Tatiana
• Granted Patent: US 9,505,867 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61L 31/048 obtained by reactions only involving carbon-to-carbon unsaturated bonds A61L31/041 takes precedence
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 41/00 Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C08F 297/02 using a catalyst of the anionic type
• CPC Class Codes: C08L 53/00 Compositions of block copolymers containing at least one sequence of a polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds; Compositions of derivatives of such polymers
• CPC Class Codes: C08L 53/005 Modified block copolymers

Abstract

The invention features a triblock copolymer including a hydrophilic block; a hydrophobic block; and a positively charged block capable of reversibly complexing a negatively charged molecule, e.g., a nucleic acid, wherein the hydrophobic block is disposed between the hydrophilic block and the positively charged block. Desirably, the triblock copolymer is capable of self-assembling into a supramolecular structure, such as a micelle or vesicle. The invention further features methods of delivering negatively charged molecules and methods of treating a disease or condition using the polymers of the invention.

Claims

1. A triblock copolymer comprising:

(i) a hydrophilic block;

(ii) a hydrophobic block; and

(iii) a positively charged block capable of reversibly complexing a negatively charged molecule,

wherein said hydrophobic block is disposed between said hydrophilic block and said positively charged block.

2. The triblock copolymer of claim 1, wherein said triblock copolymer is capable of self-assembling into a supramolecular structure.

3. The triblock copolymer of claim 2, wherein said supramolecular structure is a micelle or vesicle.

4. The triblock copolymer of claim 1, wherein the bond between said hydrophobic block and said positively charged block is labile in an endosome.

5. The triblock copolymer of claim 4, wherein said bond comprises a disulfide bond, vinyl ether, orthoester, acyl hydrazone, or a —

N—

PO₃—

group.

6. The triblock copolymer of claim 1, wherein said hydrophilic block comprises poly(ethylene glycol), poly(ethylene oxide)-co-poly(propylene oxide) di- or multiblock copolymers, poly(ethylene oxide), poly(vinyl alcohol), poly(ethylene-co-vinyl alcohol), poly(N-vinyl pyrrolidone), poly(acrylic acid), poly(ethyloxazoline), poly(alkylacrylates), poly(acrylamide), poly(N-alkylacrylamides), polypeptide, polysaccharide, poly(N,N-dialkylacrylamides), hyaluronic acid, or poly (N-acryloylmorpholine).

7. The triblock copolymer of claim 1, wherein said hydrophobic block comprises poly(propylene sulfide), poly(propylene glycol), esterified poly(acrylic acid), esterified poly(glutamic acid), esterified poly(aspartic acid), or a polypeptide.

8. The triblock copolymer of claim 1, wherein said positively charged block comprises a polypeptide, poly(ethyleneimine), or poly(amidoamine).

9. The triblock copolymer of claim 8, wherein said polypeptide comprises TAT, Arg₉, Lys₉, or Arg-Lys copolymers.

10. The triblock copolymer of claim 1, wherein said hydrophilic block is PEG, said hydrophobic block is PPS, and said positively charged block is a polypeptide.

11. The triblock copolymer of claim 10, wherein said triblock copolymer comprises PEG₄₅, PPS₅ or PPS₁₀, and TAT.

12. The triblock copolymer of claim 1, further comprising an internalization agent.

13. The triblock copolymer of claim 12, wherein said internalization agent comprises transferrin, folate, a lectin, a growth factor, an RGD peptide, or a mannose-containing glycopeptide.

14. The triblock copolymer of claim 1, further comprising a negatively charged molecule complexed to said positively charged block.

15. The triblock copolymer of claim 14, wherein said negatively charged molecule comprises a nucleic acid.

16. The triblock copolymer of claim 15, wherein said nucleic acid is siRNA, ODN, or a plasmid.

17. The triblock copolymer of claim 16, wherein said nucleic acid is siRNA that targets HIF-1α

or PAI-1.

18. A pharmaceutical composition comprising (i) a triblock copolymer of claim 1, (ii) a nucleic acid complexed to said triblock copolymer, and (iii) a pharmaceutically acceptable diluent.

19. A method for delivering a negatively charged molecule to a cell, said method comprising the steps of:

(i) providing a triblock copolymer comprising;

(1) a hydrophilic block;

(2) a hydrophobic block;

(3) a positively charged block; and

(4) a negatively charged molecule reversibly complexed to said positively charged block,

wherein said hydrophobic block is disposed between said hydrophilic block and said positively charged block; and

(ii) contacting said cell with said triblock copolymer, wherein said triblock copolymer is internalized in said cell, thereby delivering said negatively charged molecule to said cell.

20. The method of claim 19, wherein said negatively charged molecule comprises a nucleic acid.

21. The method of claim 20, wherein said nucleic acid is siRNA, ODN, or plasmid.

22. The method of claim 20, wherein said nucleic acid reduces the expression of a gene in said cell.

23. The method of claim 20, wherein said nucleic acid increases the expression of a gene in said cell.

24. A method for treating a disease or condition in a subject, said method comprising the steps of:

(i) providing a triblock copolymer comprising;

(1) a hydrophilic block;

(2) a hydrophobic block;

(3) a positively charged block; and

(4) a negatively charged molecule complexed to said positively charged block,

wherein said hydrophobic block is disposed between said hydrophilic block and said positively charged block; and

(ii) administering a therapeutically-effective amount of said triblock copolymer to said subject, wherein said triblock copolymer is internalized in a cell in said subject thereby delivering said negatively charged molecule to said cell, and thereby treating said disease or condition in said subject.

25. The method of claim 24, wherein said disease or condition is cancer.

26. The method of claim 24, wherein said disease or condition is restenosis.

27. The method of claim 24, wherein said disease or condition is scar formation.

28. The method of claim 24, wherein said disease or condition is postsurgical adhesions.

29. The method of claim 28, wherein said negatively charged molecule is siRNA that targets HIF-1α

or PAI-1.

30. The method of claim 24, wherein said negatively charged molecule comprises a nucleic acid.

31. The method of claim 30, wherein said nucleic acid is siRNA, ODN, or pDNA.

32. The method of claim 30, wherein said nucleic acid reduces the expression of a gene in said cell.

33. The method of claim 30, wherein said nucleic acid increases the expression of a gene in said cell.