Liposome composition for delivery of nucleic acid

• US 7,112,337 B2
• Filed: 12/12/2001
• Issued: 09/26/2006
• Est. Priority Date: 04/23/1999
• Status: Expired due to Term

• Current Assignee: ALZA Corporation (Johnson & Johnson)
• Original Assignee: ALZA Corporation (Johnson & Johnson)
• Inventors: Zhang, Wei-Ming, Zalipsky, Samuel, Huang, Shi-Kun
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US10/020,671
• Publication Number:
  US 20030031704A1
• Time in Patent Office: 1,749 Days
• Field of Search: 424/450, 424/1.21, 424/9.321, 424/9.51, 424/417, 428/402.2, 514/44
• US Class Current: 424/450
• CPC Class Codes: A61K 47/542 Carboxylic acids, e.g. a fatty acid or an amino acid
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/551 one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• 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: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07D 233/64 with substituted hydrocarbon radicals attached to ring carbon atoms, e.g. histidine
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

A liposome composition for delivery of a nucleic acid in vivo or ex vivo is described. The liposomes in the composition are comprised of (i) a lipid that is neutral in charge at physiologic pH and positively charged at pH values less than physiologic pH and (ii) a lipid joined to a hydrophilic polymer by a dithiobenzyl linkage. The liposomes are associated with a nucleic acid for delivery to a cell.

Claims

1. A composition for administration of a nucleic acid, comprising:

(a) liposomes comprised of

(i) a lipid having the formula

where each of R¹ and R² is an alkyl or alkenyl chain having between 8-24 carbon atoms, and each of R¹ or R² are independently selected;

n=0-20;

L is selected from the group consisting of (1) —

X—

(C═

O)—

Y—

[[CH₂—

]], (2) —

X—

(C═

O)—

, and (3) —

X—

[[CH₂—

]], where X and Y are independently selected from oxygen, NH and a direct bond;

Z is a weakly basic moiety that has a pk of less than 7.4 and greater than about 4.0; and

(ii) a compound having the general structure;

wherein R³ is a hydrophilic polymer comprising a linkage for attachment to the dithiobenzyl moiety;

R⁴ is selected from the group consisting of H, alkyl and aryl;

R⁵ is selected from the group consisting of O(C═

O)R⁷, S(C═

O)R⁷, and O(C═

S)R⁷;

R⁷ comprises an amine-containing lipid; and

R⁶ is selected from the group consisting of H, alkyl and aryl; and

where orientation of CH₂—

R⁵ is selected from the ortho position and the para position; and

(b) a nucleic acid associated with said liposomes.

2. The composition of claim 1, wherein X is NH and Y is oxygen.

3. The composition of claim 1, wherein L is a carbamate linkage, an ester linkage, or a carbonate linkage.

4. The composition of claim 1, wherein L is NH—

(C═

O)—

O—

[[CH₂]].

5. The composition of claim 1, wherein Z is an imidazole.

6. The composition of claim 1, comprising between 1-80 mole percent of the lipid.

7. The composition of claim 1, wherein Z is a moiety having a pK value between 5.0-6.5.

8. The composition of claim 1, wherein each of R¹ and R² is an unbranched alkyl or alkenyl chain having between 8-24 carbon atoms.

9. The composition of claim 8, wherein each of R¹ and R² is C₁₇H₃₅.

10. The composition of claim 1, wherein n is between 1-10.

11. The composition of claim 1, wherein R⁶ is H and R⁴ is selected from the group consisting of CH₃, C₂H₅ and C₃H₈.

12. The composition of claim 1, wherein the amine-containing lipid comprises either a single hydrocarbon tail or a double hydrocarbon tail.

13. The composition of claim 1, wherein the amine-containing lipid is a phospholipid having a double hydrocarbon tail.

14. The composition of claim 1, wherein R⁴ and R⁶ are alkyls.

15. The composition of claim 1, wherein R³ is selected from the group consisting of polyvinylpyrrolidone, polyvinylmethylether, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyloxazoline, polyhydroxypropyl-methacrylamide, polymethacrylamide, polydimethyl-acrylamide, polyhydroxypropylmethacrylate, polyhydroxyethylacrylate, hydroxymethylcellulose, hydroxyethylcellulose, polyethyleneglycol, polyaspartamide, copolymers thereof, and polyethyleneoxide-polypropylene oxide.

16. The composition of claim 1, wherein R³ is polyethyleneglycol.

17. The composition of claim 16, wherein R⁶ is H and R⁴ is CH₃ or C₂H₅.

18. The composition of claim 1, wherein said liposomes include between 5-20 mole percent of the compound.

19. The composition of claim 1, further including a therapeutic compound entrapped in the liposomes.

20. The composition of claim 1, wherein said nucleic acid is entrapped in at least a portion of said liposomes.

21. The composition of claim 20, wherein the nucleic acid is selected from DNA, RNA, fragments thereof and oligonucleotides.

22. The composition of claim 1, further including a ligand for targeting the liposomes to a target site, said ligand covalently attached to a distal end of the hydrophilic polymer R³ on said compound.

23. The composition of claim 22, wherein the ligand has binding affinity for endothelial tumor cells for internalization by such cells.

24. The composition of claim 22, wherein the ligand is selected from the group consisting of E-selectin, Her-2 and FGF.

25. The composition of claim 22, wherein said ligand is selected from the group consisting of c-erbB-2 protein product of the HER2/neu oncogene, epidermal growth factor (EGF) receptor, basic fibroblast growth receptor (basic FGF) receptor, vascular endothelial growth factor receptor, E-selectin receptor, L-selectin receptor, P-selectin receptor, folate receptor, CD4 receptor, CD19 receptor, α

β

integrin receptors, and chemokine receptors.

26. The composition of claim 1, wherein said liposomes further comprise a cationic lipid.