Polymeric oligonucleotide prodrugs

1. A method of delivering nucleic acids into a cell in a mammal, comprising administering to a mammal a compound of the formula:

X 1 -(L 2 ) n -(L 1 ) o -R 1

wherein:

R 1 is a polyalkylene oxide;

L 1 a releasable linking moiety;

L 2 is a bifunctional spacing group comprising from about 2 to about 10 carbon atoms;

X 1 is a single or double oligonucleotide residue; and

n, and o are independently a positive integer.

2. The method of claim 1 , wherein the oligonucleotide comprises a nucleotide that is selected from the group consisting of

wherein

M is O or S;

B 1 and B 2 are independently selected from the group consisting of A (adenine), G (guanine), C (cytosine), T (thymine), U (uracil) and modified bases;

R 100 and R 101 are independently selected from the group consisting of H, OR′ where R′ is H, a C 1-6 alkyl, substituted alkyls, nitro, halo and aryl.

3. The method of claim 1 , wherein said oligonucleotide contains from about 10 to about 1000 nucleotides.

4. The method of claim 2 , wherein M is S.

5. The method of claim 1 , wherein the oligonucleotide residue is a phosphorothioate oligonucleotide residue.

6. The method of claim 1 , wherein said oligonucleotide residue is an antisense oligonucleotide residue or oligodeoxynucleotide residue.

7. The method of claim 1 , wherein said oligonucleotide is selected from the group consisting of oligonucleotides with phosphorodiester backbones or phosphorothioate backbones, LNA, PNA, tricyclo-DNA, decoy ODN, ribozymes, spiegelmers, and CpG oligomers.

8. The method of claim 6 , wherein said antisense oligonucleotide has a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, wherein n of SEQ ID NO: 4 is any nucleotide.

9. The method of claim 1 , wherein R 1 is a polyalkylene oxide having a capping group A, selected from the group consisting of OH, NH 2 , SH, CO 2 H, C 1-6 alkyls, and

X 2 L 2 n L 1 o   (II)

wherein X 2 is a single stranded or double stranded oligonucleotide residue.

10. The method of claim 9 , wherein the compound comprises a formula of:

H—X 1 L 2 n L 1 o —R 1 -L 1 L 2 n —X 2 .H

wherein X 2 is a 3′ oligonucleotide or 5′ oligonucleotide.

11. The method of claim 1 , wherein L 1 is selected from the group consisting of:

wherein,

Y 1′ -Y 25′ are independently selected from the group consisting of O, S or NR 9 ;

R 6′-7′ , R 9′-13′ , R 16′-25′ , R 9 and R 27′-41′ are independently selected from the group consisting of hydrogen, C 1-6 alkyls, C 3-12 branched alkyls, C 3-8 cycloalkyls, C 1-6 substituted alkyls, C 3-8 substituted cyloalkyls, aryls, substituted aryls, aralkyls, C 1-6 heteroalkyls, substituted C 1-6 heteroalkyls, C 1-6 alkoxy, phenoxy and C 1-6 heteroalkoxy;

L 5′-12′ are independently selected bifunctional spacers;

Ar is a moiety which forms a multi-substituted aromatic hydrocarbon or a multi-substituted heterocyclic group;

c, h, k, l, r, s, v, w, v′, w′, c′, and h′ are independently selected positive integers;

a, e, g, j, t, z, a′, z′, e′ and g′ are independently either zero or a positive integer; and

b, d, f, u, b′, d′ and f′ are independently zero or one.

12. The method of claim 1 , wherein R 1 is a polyethylene glycol.

13. The method of claim 1 , wherein R 1 is selected from the group consisting of:

J-O—(CH 2 CH 2 O) n′—

J-O—(CH 2 CH 2 O) n′ —CH 2 C(O)—O—,

J-O—(CH 2 CH 2 O) n′ —CH 2 CH 2 NR 48 —,

J-O—(CH 2 CH 2 O) n′ —CH 2 CH 2 SH,

—O—C(O)CH 2 —O—(CH 2 CH 2 O) n —CH 2 C(O)—O—,

—NR 48 CH 2 CH 2 —O—(CH 2 CH 2 O) n′ —CH 2 CH 2 NR 48 —,

—SHCH 2 CH 2 —O—(CH 2 CH 2 O) n′ —CH 2 CH 2 SH—,

wherein

n′ is the degree of polymerization;

R 48 is selected from the group consisting of hydrogen, C 1-6 alkyls, C 3-12 branched alkyls, C 3-8 cycloalkyls, C 1-6 substituted alkyls, C 3-8 substituted cyloalkyls, aryls, substituted aryls, aralkyls, C 1-6 heteroalkyls, substituted C 1-6 heteroalkyls, C 1-6 alkoxy, phenoxy and C 1-6 heteroalkoxy; and

J is a capping group.

14. The method of claim 1 , wherein R 1 comprises

—O—(CH 2 CH 2 O) x —

wherein x is a positive integer selected so that the weight average molecular weight is at least about 2,000 Da to about 136,000 Da.

15. The method of claim 1 , wherein R 1 has a weight average molecular weight of from about 3,000 Da to about 100,000 Da.

16. The method of claim 1 , wherein R 1 has a weight average molecular weight of from about 5,000 Da to about 40,000 Da.

17. The method of claim 1 , wherein the compound is selected from the group consisting of:

18. The method of claim 1 , wherein the compound is selected from the group consisting of

wherein

represents an oligonucleotide and point of terminal phosphate modification; and

mPEG is CH 3 —O—(CH 2 CH 2 O) x —, wherein x is a positive integer selected from about 10 to about 2300.

19. The method of claim 1 , wherein the mammal is being treated for cancer.

20. The method of claim 1 , wherein the olignucleotide down-regulates gene expression.

21. The method of claim 1 wherein the mammal is treated with a second anticancer agent that is administered simultaneously or sequentially with the compound of claim 1 .

22. The method of claim 1 , wherein the compound is selected from the group consisting of: