Hindered ester-based biodegradable linkers for oligonucleotide delivery

• US 8,110,559 B2
• Filed: 03/12/2009
• Issued: 02/07/2012
• Est. Priority Date: 09/15/2006
• Status: Expired due to Fees

• Current Assignee: Belrose Pharma, Inc.
• Original Assignee: Enzon Pharmaceuticals Inc. (Icahn Associates Holding LLC)
• Inventors: Zhao, Hong
• Primary Examiner(s): CHU, YONG LIANG
• Application Number:
  US12/402,743
• Publication Number:
  US 20100234444A1
• Time in Patent Office: 1,062 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 19/073 with 2-deoxyribosyl as the saccharide radical
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

The invention provides hindered ester-based biodegradable linkers for the delivery of oligonucleotides in vivo, as well as method of making and using the same.

Claims

1. A compound of Formula (I)

2. The compound of claim 1, wherein R₄ and R′

₄ are independently selected oligonucleotides.

3. The compound of claim 1, wherein R₄ and R′

₄ are independently selected from the group consisting of sense oligonucleotides, antisense oligonucleotides, locked nucleic acids (LNA), short interfering RNA (siRNA), microRNA (miRNA), aptamers, peptide nucleic acid (PNA), phosphorodiamidate morpholino oligonucleotides (PMO), tricyclo-DNA, double stranded oligonucleotide (decoy ODN), catalytic RNA (RNAi), aptamers, spiegelmers, CpG oligomers and in combination.

4. The compound of claim 1, wherein R₄ and R′

₄ independently comprise ribonucleic acids, deoxyribonucleic acids, or in combination.

5. The compound of claim 1, wherein R₄ and R′

₄ are independently single stranded oligonucleotides or double stranded nucleotides.

6. The compound of claim 1, wherein R₄ and R^′

₄ independently comprise phosphorodiester backbone or phosphorothioate backbone.

7. The compound of claim 1, wherein R₁ comprises a linear, terminally branched or multi-armed polyalkylene oxide.

8. The compound of claim 7, wherein the polyalkylene oxide is selected from the group consisting of polyethylene glycol and polypropylene glycol.

9. The compound of claim 7, wherein the polyalkylene oxide is selected from the group consisting of

—

Y₇₁—

(CH₂CH₂O)_n—

CH₂CH₂Y₇₁—

,

—

Y₇₁—

(CH₂CH₂O)_n—

CH₂C(═

Y₂₂)—

Y₇₁—

,

—

Y₇₁—

C(═

Y₇₂)—

(CH₂)_a2—

Y₇₃—

(CH₂CH₂O)_n—

CH₂CH₂—

Y₇₃—

(CH₂)_a2—

C(═

Y₇₂)—

Y₇₁— and

—

Y₇₁—

(CR₇₁R₇₂)_a2—

Y₇₃—

(CH₂)_b2—

O—

(CH₂CH₂O)_n—

(CH₂)_b2—

Y₇₃—

(CR₇₁R₇₂)_a2—

Y₇₁—

,

wherein;

Y₇₁ and Y₇₃ are independently O, S, SO, SO₂, NR₇₃ or a bond;

Y₇₂ is O, S, Or NR₇₄;

R₇₁-₇₄ are independently selected from among hydrogen, C_1-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_3-19 branched alkyl, C_3-8 cycloalkyl, C_1-6 substituted alkyl, C_2-6 substituted alkenyl, C_2-6 substituted alkynyl, C_3-8 substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, C_1-6 heteroalkyl, substituted C_1-6 heteroalkyl, C_1-6 alkoxy, aryloxy, C_1-6 heteroalkoxy, heteroaryloxy, C_2-6 alkanoyl, arylcarbonyl, C_2-6 alkoxycarbonyl, aryloxycarbonyl, C_2-6 alkanoyloxy, arylcarbonyloxy, C_2-6 substituted alkanoyl, substituted arylcarbonyl, C_2-6 substituted alkanoyloxy, substituted aryloxycarbonyl, C_2-6 substituted alkanoyloxy and substituted arylcarbonyloxy;

(a2) and (b2) are independently zero or a positive integer; and

(n) is an integer from about 10 to about 2300.

10. The compound of claim 7, wherein the polyalkylene oxide is a polyethylene glycol of the formula, —

O—

(CH₂CH₂O)_n—

wherein (n) is an integer from about 10 to about 2,300.

11. The compound of claim 1, wherein R₁ has an average molecular weight from about 2,000 to about 100,000 daltons.

12. The compound of claim 1, wherein R₁ has an average molecular weight of from about 5,000 to about 60,000 daltons.

13. The compound of claim 1, wherein R₁ has an average molecular weight from about 5,000 to about 25,000 daltons or from about 20,000 to about 45,000 daltons.

14. The compound of claim 1 selected from the group consisting of:

15. The compound of claim 1 wherein R₄ and R′

₄ are independently selected from the group consisting of SEQ ID NO;

1, SEQ ID NOs;

2 and 3, SEQ ID NO;

4, and SEQ ID NO;

5.

16. A method of preparing a hindered acyl or ester moiety-containing polymeric conjugate of Formula (I) of claim 1 comprising:

reacting a compound of Formula (VI);