N-(.omega., (.omega.-1)-dialkyloxy)- and N-(.omega., (.omega.-1)-dialkenyloxy)-alk-1-yl-N,N,N-tetrasubstituted ammonium lipids and uses therefor

• US 5,208,036 A
• Filed: 11/16/1990
• Issued: 05/04/1993
• Est. Priority Date: 01/07/1985
• Status: Expired due to Term

• Current Assignee: Syntec Incorporated (Atos SE)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Felgner, Philip L., Gadek, Thomas R., Roman, Richard B., Jones, Gordon H., Eppstein, Deborah A.
• Primary Examiner(s): Page, Thurman K.
• Assistant Examiner(s): AZPURU, CARLOS A
• Application Number:
  US07/614,412
• Time in Patent Office: 900 Days
• Field of Search: 424/422, 424/423, 424/427, 424/428, 424/449, 424/450, 435/829, 264/4.1, 264/4.33, 264/4.6
• US Class Current: 424/450
• CPC Class Codes: A61K 47/186 Quaternary ammonium compounds, e.g. benzalkonium chloride or cetrimide
• 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: C07D 453/02 containing not further condensed quinuclidine ring systems
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: Y10S 435/829 Alcaligenes

Abstract

This invention relates to compounds of the formula ##STR1## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms; R³, R⁴ and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R³, R⁴, and R⁵ are taken together to form quinuclidino, piperidino, pyrrolidino, or morpholino; n is 1 to 8; and X is a pharmaceutically acceptable anion.

Claims

1. A polyanion-lipid complex formed from a compound of Formula I and a polyanion, wherein Formula I is represented by the formula:

##STR8## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms;

R³, R⁴, and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms or when two or three of R³, R⁴, and R⁵ are taken together to form quinoclidino, pyrrolidono, piperidino or morpholino;

n is 1 to 8; and

X is a pharmaceutically acceptable atom; and

an aqueous solution in a quantity sufficient to make 100% by volume.

2. The complex of claim 1 wherein the polyanion is a polynucleotide.

3. The complex of claim 2 wherein the compound of Formula I and the polyanion are represented in the complex in a charge ratio such that the complex has a net positive charge.

4. The complex of claim 3 wherein the charge ratio is in the range of 1000:

1 to 1;

1.

5. The complex of claim 3 wherein the charge ratio is in the range of 20:

1 to 1;

1.

6. The complex of claim 3 wherein the charge ratio is in the range of 5:

1 to 2;

1.

7. The complex of claim 3 wherein the polyanion is DNA.

8. The complex of claim 3 wherein the polyanion is RNA.

10. The complex of claim 3 wherein the polyanion is a polypeptide.

9. The complex of claim 9 wherein the RNA is double stranded.

11. A method for forming a polyanion-lipid complex, said method comprising the step of contacting a liposomal composition prepared from a compound of Formula I with a negatively charged polyanion, wherein Formula I is represented by the formula:

##STR9## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms;

R³, R⁴, and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms or when two or three of R³, R⁴, and R⁵ are taken together to form quinoclidino, pyrrolidono, piperidino or morpholino;

n is 1 to 8; and

X is a pharmaceutically acceptable atom and an aqueous solution in a quantity sufficient to make 100% by volume.

12. The method of claim 11 wherein the polyanion is a polynucleotide.

13. The method of claim 11 wherein the polyanion is a polypeptide.

14. The method of claim 11 wherein the positively charged liposome-forming lipid and the polyanion are represented in the complex in a charge ratio such that the complex has a net positive charge.

15. The method of claim 11 wherein the ratio is in the range of 5:

1 to 2;

1.

16. The method of claim 12 wherein the polynucleotide is DNA.

17. The method of claim 12 wherein the polynucleotide is RNA.

18. A positively-charged polynucleotide-liposome complex, comprising:

a polynucleotide; and

a lipid of the Formula;

##STR10## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms;

R³, R⁴ and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R³, R⁴, and R⁵ are taken together to form quinculidino, pyrrolidino, piperidino, or morpholino;

n is 1 to 8; and

X is a pharmaceutically acceptable anion.

19. A complex according to claim 18 wherein R¹ and R² are the same and are alkyl of 10 to 20 carbon atoms, R³, R⁴, and R⁵ are methyl or ethyl, n is 1 to 4 and X is a halide ion.

20. A complex according to claim 19 wherein n is 1.

21. The complex of claim 20, wherein the compound is (±

) N-(2,3-di-(9-(Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammonium chloride or an optical isomer thereof.

22. A method for preparing a positively-charged polynucleotide-lipid complex, said method comprising the steps of:

forming a liposome from a positively charged lipid of Formula I ##STR11## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms;

R³, R⁴ and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R³, R⁴, and R⁵ are taken together to form quinuclidino, pyrrolidino, piperidino, or morpholino;

n is 1 to 8; and

X is a pharmaceutically acceptable anion; and

contacting the liposome with a polynucleotide to obtain a polynucleotide-lipid complex.

23. A method according to claim 22 wherein R¹ and R² are the same and are alkyl of 10 to 20 carbon atoms, R³, R⁴, and R⁵ are methyl or ethyl, n is 1 to 4 and X is a halide ion.

24. A method according to claim 23 wherein n is 1.

25. The method of claim 24, wherein the compound of Formula I is (±

) N-(2,3-di-(9-(Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammonium chloride or an optical isomer thereof.

26. The method of claim 22, comprising the further step of providing, for the contacting step, a quantity of liposome and a quantity of polynucleotide equivalent to a molar ratio of the compound of Formula I to the polynucleotide in the range of from 5:

1 to 1;

1.

27. A positively-charged polynucleotide liposome complex, comprising:

a polynucleotide

a lipid of the Formula;

##STR12## or an optical isomer thereof wherein R¹ and R² are the same or different and are an alkyl or alkenyl group of 6 to 24 carbons;

R³ R⁴ and R⁵ are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R³, R⁴ and R⁵ are taken together to form quinoclidino, pyrrolidino, piperidino, or morpholino;

n is 1 to 8; and

x is a pharmaceutically acceptable anion.

28. A complex according to claim 27 wherein R¹ and R² are the same and are alkyl of 10 to 20 carbon atoms, R³, R⁴ and R⁵ are methyl or ethyl, n is 1 to 4 and X is a halide ion.

29. A complex according to claim 28 wherein n is 1.

30. The complex of claim 28 wherein the lipid is (±

) N-(2,3-di(9-(Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammonium chlorie or an optical isomer thereof.