Modified polymers for delivery of polynucleotides, method of manufacture, and methods of use thereof

• US 8,349,308 B2
• Filed: 03/28/2011
• Issued: 01/08/2013
• Est. Priority Date: 03/26/2010
• Status: Expired due to Fees

• Current Assignee: Mersana Therapeutics, Inc.
• Original Assignee: Mersana Therapeutics, Inc.
• Inventors: Yurkovetskiy, Aleksandr, Yin, Mao, Lowinger, Timothy B., Cabral, Carolina B., Hammond, Charles E., Stevenson, Cheri A.
• Primary Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US13/073,815
• Publication Number:
  US 20110243880A1
• Time in Patent Office: 652 Days
• Field of Search: None
• US Class Current: 424/78.17
• CPC Class Codes: A61K 47/59 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: C08L 59/00 Compositions of polyacetals; Compositions of derivatives of polyacetals of polyvinyl acetals C08L29/14
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/12 catalytic nucleic acids, e.g. ribozymes
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

A polynucleotide delivery vehicle comprising a modified polymer is provided herein, the modified polymer having the following formula:

in which W₁, W₂, W₃, W₄, W₅, W₆, Z₁, Z₂, Z₃, Z₄, Z₅, Z₉, R₁, R₂, R₃, R₄, R₅, n₁, n₂, n₃, n₄, n₅, and n₆ are defined herein. Also disclosed are methods of delivering a polynucleotide to the cytoplasm of a selected tissue type or cell type and methods of reducing expression of a gene in a cell or a subject in need thereof with the modified polymer.

Claims

1. A modified polyacetal of Formula (VI):

2. The modified polyacetal of claim 1, wherein

m₁ is 0.002 to 0.25;

m₃ is 0.002 to 100;

m₄ is 0.03 to 0.30; and

m₅ is 0.01 to 0.03.

3. The modified polyacetal of claim 1, wherein when Z₉ is Z₆-T₁,

(i) n₁ is not 0 and each of n₂, n₃, n₄, and n₅ is 0;

(ii) neither n₁ nor n₂ is 0 and each of n₃, n₄, and n₅ is 0;

(iii) none of n₁, n₂ and n₃ is 0 and each of n₄ and n₅ is 0;

(iv) none of n₁, n₂ and n₄ is 0 and each of n₃ and n₅ is 0;

(v) none of n₁, n₂, n₃ and n₄ is 0 and n₅ is 0;

(vi) neither n₁ nor n₄ is 0 and each of n₂, n₃ and n₅ is 0;

(vii) n₂ is not 0 and each of n₁, n₃, n₄, and n₅ is 0;

(viii) neither n₂ nor n₃ is 0 and each of n₁, n₄ and n₅ is 0;

(ix) neither n₂ nor n₄ is 0 and each of n₁, n₃ and n₅ is 0;

(x) none of n₂, n₃ and n₄ is 0 and each of n₁ and n₅ is 0;

(xi) none of n₁, n₂, n₃, n₄, and n₅ is 0;

or

(xii) each of n₁, n₂, n₃, n₄, and n₅ is 0.

4. The modified polyacetal of claim 1, wherein when Z₉ is Z₆-T₁,

n is between about 0.01 and about 0.9996 inclusive;

n₁ is between about 0.002 and about 0.25 inclusive;

n₂ is between about 0.02 and about 0.90 inclusive;

n₃ is between about 0.02 and about 0.81 inclusive;

n₄ is between about 0.03 and about 0.30 inclusive;

n₅ is between about 0.01 and about 0.03 inclusive; and

n₆ is between about 0.0004 and about 0.10 inclusive.

5. The modified polyacetal of claim 1, wherein when Z₉ is Z₈, each of n₁, n₃, n₄, and n₅ is 0, and R₂ is a linear or branched polyamino moiety optionally substituted with one or more substituents selected from the group consisting of -Q-R₁, -Q-R₃, -Q-R₄, and -Q-R₅.

6. The modified polyacetal of claim 1, wherein when Z₉ is Z₈,

(i) m₁ is not 0 and each of m₃, m₄ and m₅ is 0;

(ii) neither m₁ nor m₄ is 0 and each of m₃ and m₅ is 0;

(iii) none of m₁, m₄ and m₅ is 0 and m₃ is 0;

(iv) neither m₁ nor m₃ is 0 and each of m₄ and m₅ is 0;

(v) none of m₁, m₃ and m₄ is 0 and m₅ is 0;

or

(vi) none of m₁, m₃, m₄ and m₅ is 0.

7. The modified polyacetal of claim 1, wherein when Z₉ is Z₈,

n is between about 0.70 and about 0.99 inclusive;

each of n₁, n₃, n₄, and n₅ is 0;

m₁ is 0.002 to 0.25;

m₃ is 0.002 to 100;

m₄ is 0.03 to 0.30;

m₅ is 0.01 to 0.03; and

m₆ is 0.0004 to 0.10.

8. The modified polyacetal of claim 1, wherein each of Z₁, Z₂, Z₃, Z₄, Z₅, and Z₆, independently, is ethylenediamine, piperazine, bis(piperidine), 1,3-diaminopropane, 1,4-diaminobutane, 1,5-diaminopentane, decamethylenediamine, hexamethylenediamine, lysine, histidine, arginine, tryptophan, agmatine or ornithine.

9. The modified polyacetal of claim 8, wherein each of Z₁, Z₂, Z₃, Z₄, Z₅, and Z₆ is

10. The modified polyacetal of claim 1, wherein Z₇ is:

11. The modified polyacetal of claim 10, wherein Z₇ is:

12. The modified polyacetal of claim 1, wherein Z₈, when otherwise unsubstituted, is:

13. The modified polyacetal of claim 12, wherein Z₈, when otherwise unsubstituted, is:

(1) a linear polyethylenimine having a molecular weight of about 500 to about 25000 dalton;

(2) a branched polyethylenimine having a molecular weight of about 500 to about 25000 dalton;

14. The modified polyacetal of claim 13, wherein Z₈, when otherwise unsubstituted, is a linear polyethylenimine having a molecular weight of about 500 to about 2500 dalton or a branched polyethylenimine having a molecular weight of about 500 to about 1200 dalton.

15. The modified polyacetal of claim 12, wherein each of R_y and R_z, independently, is a polyamino moiety comprising a monomer unit of —

[C_2-6 alkyl-NH]—

.

16. The modified polyacetal of claim 1, wherein R₁ comprises galactosamine, galactose, N-acetylgalactosamine, folic acid, RGD peptides, LHRH receptor targeting peptides, ErbB2 (HER2) receptor targeting peptides, prostate specific membrane bound antigen (PSMA) targeting peptides, lipoprotein receptor LRP1 targeting ligands, ApoE protein derived peptides or transferrin.

17. The modified polyacetal of claim 1, wherein R₂ is:

18. The modified polyacetal of claim 1, wherein R₃ is of Formula (XVI):

19. The modified polyacetal of claim 18, wherein R₃ is:

20. The modified polyacetal of claim 1, wherein R₄ comprises C_5-20 saturated or unsaturated fatty acids, C_6-22 alkylamines, cholesterol, cholesterol derivatives or amino containing lipids.

21. The modified polyacetal of claim 20, wherein R₄ is:

22. The modified polyacetal of claim 1, wherein R₆ is a natural, synthetic, or semi-synthetic polynucleotide, DNA, RNA or an oligonucleotide.

23. The modified polyacetal of claim 22, wherein R₆ a double stranded oligonucleotide having about 12 to about 30 nucleotides or a single stranded oligonucleotide having about 8 to about 64 nucleotides.

24. The modified polyacetal of claim 1, wherein the polyacetal backbone has a molecular weight of about 100 kDa, about 70 kDa, about 60 kDa or about 40 kDa.

25. The modified polyacetal claim 1, wherein R₂ is

26. The modified polyacetal claim 1, wherein R₃ is

27. The modified polyacetal of claim 1, wherein R₄ is

28. A method for delivering a polynucleotide to the cytoplasm of a selected tissue type or cell type, comprising contacting the modified polyacetal of claim 1 with the selected tissue type or cell type.

29. The method of claim 28, wherein the tissue type is a liver tissue or a kidney tissue and the cell type is a blood cell, an endothelial cell, a cancer cell, a pancreatic cell, or a neural cell.

30. A method of reducing expression of a gene in a cell, comprising:

delivering to the cytoplasm of a cell an effective amount of the modified polyacetal of claim 1, wherein the modified polyacetal contains a polynucleotide that is complementary to at least a portion of the gene.

31. A method of reducing expression of a gene in a subject comprising:

administering to a subject in need thereof an effective amount of the modified polyacetal of claim 1, wherein the modified polyacetal contains a polynucleotide that is complementary to at least a portion of the gene.