Transfection reagents

Patent metadata

Field	Value
Publication number	US-9358300-B2
Application number	US-201414336887-A
Country	US
Kind code	B2
Filing date	Jul 21, 2014
Priority date	Nov 12, 1998
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

Disclosed are compounds capable of facilitating transport of biologically active agents or substances into cells having the general structure:

wherein Q is selected from the group consisting of N, O and S; L is any bivalent organic radical capable of linking each Q, such as C, CH, (CH 2 ) l , or {(CH 2 ) i —Y—(CH 2 ) j } k , wherein Y is selected from the group consisting of CH 2 , an ether, a polyether, an amide, a polyamide, an ester, a sulfide, a urea, a thiourea, a guanidyl, a carbamoyl, a carbonate, a phosphate, a sulfate, a sulfoxide, an imine, a carbonyl, and a secondary amino group and wherein Y is optionally substituted by —X 1 -L′-X 2 —Z or —Z; R 1 R 6 , independently of one another, are selected from the group consisting of H, —(CH 2 ) p -D-Z, an alkyl, an alkenyl, an aryl, and an alkyl or alkyl ether optionally substituted by one or more of an alcohol, an aminoalcohol, an amine, an amide, an ether, a polyether, a polyamide, an ester, a mercaptan, an alkylthio, a urea, a thiourea, a guanidyl, or a carbamoyl group, and wherein at least one of R 1 , R 3 , R 4 and R 6 is a straight chain or branched, cyclic, alkyl, alkenyl, alkynyl or aryl group; and anyone of R 1 , R 3 , R 4 and/or R 6 may optionally be covalently linked with each other, with Y or with L when L is C or CH to form a cyclic moiety; Z is selected from the group consisting of amine, spermiyl, carboxyspermiyl, guanidyl, spermidinyl, putricinyl, diaminoalkyl, pyridyl, piperidinyl, pyrrolidinyl, polyamine, amino acid, peptide, and protein; X 1 and X 2 , independently of one another, are selected from the group consisting of NH, O, S, alkylene, and arylene; L′ is selected from the group consisting of alkylene, alkenylene, alkynylene, arylene, alkylene ether, and polyether; D is Q or a bond; A 1 and A 2 , independently of one another, are selected from the group consisting of CH 2 O, CH 2 S, CH 2 NH, C(O), C(NH), C(S) and (CH 2 )t; X is a physiologically acceptable anion; m, n, r, s, u, v, w and y are 0 or 1, with the proviso that when both m and n are 0 at least one of r, s, u and y is other than 0; i, j, k, l, p and are integers from 0 to about 100; q is an integer from 1 to about 1000; and a is the number of positive charge divided by the valence of the anion.

First claim

We claim:

1. A transfection complex formed in an aqueous medium, said transfection complex comprising:
an expressible nucleic acid;
a compound having the structure:

  or a polycation thereof;
wherein R 4 is a straight-chain, branched or cyclic, alkyl or alkenyl group having 8 to 24carbon atoms, R 5 is an alkyl group substituted with one or more amines and one or more alcohols, and R 7 is H or a carbohydrate; and

at least one protein or peptide transfection enhancer.

2. The transfection complex according to claim 1 , wherein the protein or peptide transfection enhancer enhances cell-targeting, uptake, internalization, nuclear targeting or expression of the expressible nucleic acid.

3. The transfection complex according to claim 1 , wherein the protein or peptide transfection enhances uptake of the expressible nucleic acid.

4. The transfection complex according to claim 1 , wherein R 5 is an alkyl group substituted with one amine and one or more alcohols.

5. The transfection complex according to claim 1 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

6. The transfection complex according to claim 5 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

7. The transfection complex according to claim 1 , wherein R 5 is an alkyl group having 3-30 carbon atoms and is substituted by one amine and one or more alcohols.

8. The transfection complex according to claim 7 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

9. The transfection complex according to claim 7 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

10. The transfection complex according to claim 2 , wherein the protein or peptide transfection enhancer enhances uptake, internalization or nuclear targeting of the expressible nucleic acid.

11. The transfection complex according to claim 10 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

12. The transfection complex according to claim 1 , wherein R 7 is H.

13. The transfection complex according to claim 12 , wherein R 4 is a straight-chain alkyl group having 8 to 24 carbon atoms.

14. The transfection complex according to claim 13 , wherein R 5 is an alkyl group having 3-30 carbon atoms and is substituted with one amine and one or more alcohols.

15. The transfection complex according to claim 14 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

16. The transfection complex according to claim 14 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

17. The transfection complex according to claim 14 , wherein R 5 is a propyl or a butyl group which is substituted by one amine and one or more alcohols.

18. The transfection complex according to claim 17 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

19. The transfection complex according to claim 18 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

20. The transfection complex according to claim 1 , wherein R 4 is a Straight-chain alkyl group having 8 to 24 carbon atoms, R 5 is a butyl group substituted with one amine and one or more alcohols, and R 7 is H.

21. The transfection complex according to claim 20 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

22. The transfection complex according to claim 21 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

23. The transfection complex according to claim 1 , wherein R 4 is a straight-chain alkyl group having 8 to 24 carbon atoms, R 5 is a propyl group substituted with one amine and one alcohol, and R 7 is H.

24. The transfection complex according to claim 23 , wherein the amine substituent of the R 5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group.

25. The transfection complex according to claim 24 , wherein the amine substituent of the R 5 alkyl group has the structure:

wherein n is an integer from 7-23.

26. The transfection complex according to claim 1 , wherein the transfection complex further comprises one or more neutral lipids.

27. The transfection complex according to claim 26 , wherein the neutral lipid is DOPE, DOPC or cholesterol.

28. The transfection complex according to claim 1 , wherein the transfection complex comprises a liposome.

Assignees

• Life Technologies Corp

Inventors

• Chu Yongliang
• Masoud Malek
• Gebeyehu Gulilat

Classifications

• A61K31/14 Primary

  Quaternary ammonium compounds, e.g. edrophonium, choline (betaines A61K31/205) · CPC title

• A61K38/28

  Insulins · CPC title

• A61K47/48046 Primary

  Human Necessities · mapped topic

• C07C211/21

  Monoamines · CPC title

• C07D209/48

  with oxygen atoms in positions 1 and 3, e.g. phthalimide · CPC title