TRANSFECTION REAGENTS

• US 20120238747A1
• Filed: 03/14/2012
• Published: 09/20/2012
• Est. Priority Date: 11/12/1998
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: CHU, Yongliang, Masoud, Malek, Gebeyehu, Gulilat
• Granted Patent: US 8,785,200 B2
• Time in Patent Office: Days
• US Class Current: 540/467
• CPC Class Codes: A61K 31/14 Quaternary ammonium compounds, e.g. edrophonium, choline betaines A61K31/205
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 38/40 Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 9/16 Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction A61K9/20 takes precedence if the final form is a tablet; microspheres with drug-free outer coating, microcapsules A61K9/50; mixture of different granules, microcapsules, (coated) microparticles A61K9/5084; nanoparticles A61K9/51
• CPC Class Codes: C07C 211/21 Monoamines
• CPC Class Codes: C07C 211/63 having quaternised nitrogen atoms bound to acyclic carbon atoms
• CPC Class Codes: C07C 211/64 having quaternised nitrogen atoms bound to carbon atoms of six-membered aromatic rings
• CPC Class Codes: C07C 229/26 having more than one amino group bound to the carbon skeleton, e.g. lysine
• CPC Class Codes: C07D 209/48 with oxygen atoms in positions 1 and 3, e.g. phthalimide
• CPC Class Codes: C07D 241/12 with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: Y10S 435/81 Packaged device or kit

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, (CH₂)l, or {(CH₂)i-Y—(CH₂)j}k, wherein Y is selected from the group consisting of CH₂, 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 —X₁-L′-X₂—Z or —Z; R₁-R₆, independently of one another, are selected from the group consisting of H, —(CH₂)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 R₁, R₃, R₄ and R₆ is a straight chain or branched, cyclic, alkyl, alkenyl, alkynyl or aryl group; and anyone of R₁, R₃, R₄ and/or R₆ 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; X₁ and X₂, independently of one another, are selected from the group consisting of NH, O, S, alkylene, and arylene; U is selected from the group consisting of alkylene, alkenylene, alkynylene, arylene, alkylene ether, and polyether; D is Q or a bond; A₁ and A₂, independently of one another, are selected from the group consisting of CH₂O, CH₂S, CH₂NH, C(O), C{NH), C(S) and (CH₂)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.

Claims

1. A compound or polycation having the formula:

2. The compound or polycation of claim 1 wherein at least two of R₁, R₃, R₄ and R₆ are a straight-chain, branched, or cyclic alkyl, alkenyl, alkynyl or aryl groups having from about 6 to about 64 carbon atoms.

3. The compound of claim 1 wherein at least two of R₁, R₃, R₄ and R₆ are a straight-chain, branched, or cyclic alkyl, alkenyl, alkynyl or aryl groups having from about 8 to about 24 carbon atoms.

4. The compound of polycation of claim 1 wherein Q is N.

5. The compound or polycation of claim 1 wherein L is C, CH, (CH₂)_l or {(CH₂)_i—

Y—

(CH₂)_j}_k, where Y is selected from the group consisting of CH₂, 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 —

X₁-L′

-X₂—

Z or —

Z;

X₁ and X₂, 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;

A₁ and A₂ independently of one another, are selected from the group consisting of CH₂O, CH₂S, CH₂NH, C(O), C(NH), C(S) and (CH₂)_t;

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 t are integers from 1 to about 100; and

q is an integer from 1 to about 1000.

6. The compound or polycation of claim 1 where L is (CH2)l or {(CH2)i-Y—

(CH2)j}k, wherein Y is selected from the group consisting of CH2, an ether, a polyether, an amide, a polyamide, an ester, a sulfide, a urea, a thiourea, a guanidyl, a carbamoyl, a carbonate, and a secondary amino group; and

l, j and k are integers from 1 to 4.

7. The compound of claim 1 wherein at least two of R₁-R₆ are —

(CH₂)_p-D-Z groups.

8. The compound or polycation of claim 1 wherein Z is selected from the group consisting of spermiyl, spermidiyl, amino acid, peptidyl, diaminoalkyl, and polyamine when Z is part of any of the groups —

(CH₂)bNHC(O)Z, —

(CH₂)cNHC(O)Z, —

CH₂CH(OR₇)CH₂NHC(O)Z, or —

CH₂CH(OR₈)CH₂NHC(O)Z.

9. The compound or polycation of claim 1 having the formula:

10. The compound or polycation of claim 9 wherein at least two of R₁, R₃, R₄ and R₆ are a straight-chain, branched or cyclic alkenyl, alkynyl, or aryl group.

11. The compound or polycation of claim 19 wherein l is 4.

12. The compound of claim 1 having the formula:

13. The compound of polycation of claim 12 wherein R₁ and R₄ are straight-chain, branched or cyclic alkyl, alkenyl, alkynyl or aryl groups having from about 8 to about 24 carbon atoms.

14. The compound or polycation of claim 1 wherein R3 and R6 are alkyl amine groups.

15. The compound or polycation of claim 1 wherein R₃ and R₆ are

16. The compound of polycation of claim 1 wherein R₃ and R₆ are

17. The compound or polycation of claim 1 wherein R₃ and R₆ are

18. The compound or polycation of claim 1 wherein R₃ and R₆ are

19. The compound or polycation of claim 1 wherein R₃ and R₆ are

20. The compound or polycation of claim 1 wherein R₃ and R₆ are