Cationic lipids for intracellular delivery of biologically active molecules

• US 5,264,618 A
• Filed: 04/16/1991
• Issued: 11/23/1993
• Est. Priority Date: 04/19/1990
• Status: Expired due to Term

• Current Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Original Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Inventors: Kumar, Raj, Felgner, Philip L., Basava, Channa, Hwang-Felgner, Jiin-Yu, Border, Richard C.
• Primary Examiner(s): Killos, Paul J.
• Application Number:
  US07/686,746
• Time in Patent Office: 952 Days
• Field of Search: 560/224, 554/227, 554/226, 554/223, 530/323, 574/292, 574/549, 574/552
• US Class Current: 560/224
• CPC Class Codes: A61K 47/186 Quaternary ammonium compounds, e.g. benzalkonium chloride or cetrimide
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• 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 31/12 Antivirals
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

Disclosed are cationic lipids capable of facilitating transport of biologically active agents into cells, including the transfection of cells by therapeutic polynucleotides, the delivery of antiviral drugs, and the introduction of immunogenic peptides. The cationic lipids, comprising an ammonium group, have the general structure ##STR1## Also disclosed are adducts of these compounds comprising additional cationic sites that enhance the transfective or transport activity. Structure-activity correlations provide for the selection of preferred compounds to be synthesized for this purpose. Compositions disclosed for use of these cationic lipid include formulations for in vitro transfection and pharmaceutical formulations for parenteral and topical administration of therapeutic agents.

Claims

1. A composition having the structure ##STR15## wherein Y¹ and Y² are the same or different and are, --O--C(O)--, or --O--;

R¹ is H, or C₁ to C₂₄ alkyl or alkenyl;

R² is C₁ to C₂₄ alkyl or alkenyl;

R³ and R⁴ are the same or different and are C₁ to C₂₄ alkyl, or H;

R⁵ is C₁ to C₂₄ alkyl straight chain or branched chain;

R⁶ is --C(O)--(CH₂)_m --NH--, a diaminocarboxylate ester group which is alkyl, aryl, or aralkyl, or --C(O)--(CH₂)_m --NH-- linked to said diaminocarboxylate ester group, or is absent;

R⁷ is H, spermine, spermidine, a histone, or a protein with DNA-binding specificity, or the same groups wherein the amine functionalities of the R⁷ moiety are quaternized with R³, R⁴, or R⁵ groups;

or

R⁷ is an L- or D-alpha amino acid having a positively charged group on the side chain, said amino acids comprising arginine, histidine, lysine or ornithine or analogues thereof, or wherein the amine of the R⁷ moiety is quaternized with R³, R⁴ or R⁵ groups;

or

R⁷ is a polypeptide selected from the group consisting of L-or D-alpha amino acids, wherein at least one of the amino acids residues comprises arginine, histidine, lysine, ornithine, or analogues thereof;

n is 1 to 8;

m is 1 to 18; and

X is a non-toxic anion.

2. A composition according to claim 1 wherein R³ and R⁴ are individually C₁ to C₂₃ alkyl groups, R⁵ is --(CH₂)_m --, R⁶ is absent, R⁷ is H, and R¹ and R² individually have from 0 to 6 sites of unsaturation, and have the structure

3. A composition according to claim 2, wherein Y¹ and Y² are alike and are --O--C(O)--.

4. A composition according to claim 3, which is DL-1,2-dioleoyl-3-dimethylaminopropyl-β

-hydroxyethylammonium and its salts.

5. A composition according to claim 2 wherein Y¹ and Y² are alike and are --O--.

6. A composition according to claim 5, which is 1,2-O-dioleyl-3-dimethylaminopropyl-β

-hydroxyethylammonium and its salts.

7. A composition according to claim 2 wherein Y¹ and Y² are different and are either --O-- or --O--C(O)--.

8. A composition according to claim 7 which is 1-O-oleyl-2-oleyl-3-dimethylaminopropyl-β

-hydroxyethylammonium and its salts.

9. 3,5-(N,N-dilysyl)-diaminobenzoyl-3-(DL-1,2-dioleoyl-dimethylaminopropyl-β

-hydroxyethylamine).

10. 3,5-(N,N-dilysyl)-diaminobenzoylglycyl-3-(DL-1,2-dioleoyl-dimethylaminopropyl-β

-hydroxyethylamine).

11. L-spermine-5-carboxyl-3-(DL-1,2-dioleoyldimethylaminopropyl-β

-hydroxyethylamine).

12. A composition having the structure ##STR16## or an optical isomer thereof, wherein Y¹ and Y² are different and are either, --O--C(O)-- or --O--;

R¹ is C₁ to C₂₄ alkyl or alkenyl, or H;

R² is C₁ to C₂₄ alkyl or alkenyl;

R³, R⁴ and R⁵ are the same or different and are H, C₁ to C₁₄ alkyl, C₇ to C₁₁ aryl or alkaryl, or at least two of R³, R⁴ and

R⁵ are taken together to form quinuclidino, piperidino, pyrrolidino, or morpholino; and

X is a non-toxic anion.