Neutral nanotransporters

• US 10,138,485 B2
• Filed: 09/22/2009
• Issued: 11/27/2018
• Est. Priority Date: 09/22/2008
• Status: Active Grant

• Current Assignee: Phio Pharmaceuticals Corp.
• Original Assignee: Rxi Pharmaceuticals Corporation
• Inventors: Khvorova, Anastasia, Salomon, William, Kamens, Joanne, Samarsky, Dmitry, Woolf, Tod M., Miller, Michelle, Bulock, Karen G.
• Primary Examiner(s): Audet, Maury A
• Application Number:
  US13/120,341
• Publication Number:
  US 20110237522A1
• Time in Patent Office: 3,353 Days
• Field of Search: None
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/3231 : having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 : 5-Methylcytosine
• CPC Class Codes: C12N 2310/3515 : Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3519 : Fusion with another nucleic acid
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2320/32 : Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/51 : modulating the chemical stability, e.g. nuclease-resistance
• CPC Class Codes: C12N 2320/53 : reducing unwanted side-effects

Abstract

Neutral lipid formulations for nucleic acid delivery are provided according to the invention. The neutral lipid formulations include hydrophobically modified polynucleotides and fat mixtures. Methods of using the neutral lipid formulations are also provided.

Claims

1. A composition comprising:

a hydrophobic modified polynucleotide, wherein the hydrophobic modified polynucleotide comprises an isolated double stranded nucleic acid molecule comprising a guide strand and a passenger strand, wherein the isolated double stranded nucleic acid molecule includes a double stranded region and a single stranded region, wherein the double stranded region is from 8-14 nucleotides long, wherein the single stranded region is at the 3′

end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 phosphorothioate modifications, wherein at least 40% of the nucleotides of the isolated double stranded nucleic acid molecule are modified, and wherein a hydrophobic conjugate is attached to the isolated double stranded nucleic acid molecule at the 3′

end of the sense strand, and wherein the isolated double stranded nucleic acid molecule does not form a hairpin;

a neutral fatty mixture comprising at least 20% dioleoylphosphatidylcholine (DOPC) or distearoylphosphatidylcholine (DSPC), and at least 20% sterol; and

optionally a cargo molecule,

wherein the hydrophobic modified polynucleotide and the neutral fatty mixture forms a micelle.

2. The composition of claim 1, wherein the polynucleotide is RNA.

3. The composition of claim 1, wherein the neutral fatty mixture comprises at least 25% DOPC or 25% DSPC.

4. The composition of claim 3, wherein the neutral fatty mixture comprises at least 25% sterol.

5. The composition of claim 1, wherein the composition includes at least 20% DOPC and at least 20% cholesterol.

6. A method of inducing RNAi in a cell comprising:

contacting a cell with a composition of claim 1, wherein the polynucleotide has at least a region of sequence correspondence to a target gene and wherein the polynucleotide induces RNAi of mRNA of the target gene.

7. A method of inducing RNAi in a subject in need thereof comprising:

administering to the subject an effective amount for inducing RNAi of mRNA of a target gene, a composition of claim 1, wherein the polynucleotide has at least a region of sequence correspondence to the target gene.

8. The method of claim 7, wherein the subject is a human.

9. A method for delivering a polynucleotide to a target tissue of a subject in need thereof, comprising

administering to the subject a composition of claim 1 wherein the composition includes a fatty acid in the neutral fatty mixture or cargo molecule associated with targeting to the target tissue in order to deliver the polynucleotide to the target tissue in an amount effective to deliver the polynucleotide to the target tissue.

10. A method of inducing RNAi in a subject in need thereof comprising:

administering to the subject an effective amount for inducing RNAi of mRNA of a target gene, a composition of claim 1, wherein the polynucleotide has at least a region of sequence correspondence to the target gene, wherein the step of administering is systemic, intravenous, intraperitoneal, intradermal, topical, intranasal, inhalation, oral, intramucosal or intraocular.

11. The composition of claim 1, wherein each of the at least 40% of nucleotides independently comprises a chemical modification selected from the group consisting of 2′

d, 2′

OMe, 2′

F, hydrophobic and phosphorothioate.

12. The composition of claim 1, wherein the cargo molecule is an intralipid or a nutralipid.

13. The composition of claim 1, wherein the cargo molecule is a fatty acid mixture containing more than 74% linoleic acid, a fatty acid mixture containing at least 6% of cardiolipin, or a fatty acid mixture containing at least 74% linoleic acid and at least 6% of cardiolipin.

14. The composition of claim 1, wherein the cargo molecule is a fusogenic lipid.

15. The composition of claim 14, wherein the fusogenic lipid is DOPE.