Stable nanolipoprotein particles and related compositions methods and systems

Patent metadata

Field	Value
Publication number	US-12226529-B2
Application number	US-201615755018-A
Country	US
Kind code	B2
Filing date	Aug 25, 2016
Priority date	Aug 25, 2015
Publication date	Feb 18, 2025
Grant date	Feb 18, 2025

Abstract

Nanolipoprotein particles having at least a scaffold protein component and a membrane lipid component and related compositions, methods and systems are described. The membrane lipid component includes at least one or more membrane forming lipids, one or more polymerized lipids and/or one or more polymerizable lipids.

First claim

The invention claimed is:

1. A nanolipoprotein particle comprising:
a membrane forming lipid, a polymerized lipid and a scaffold protein,

the membrane forming lipid and the polymerized lipid arranged in a membrane forming lipid discoidal bilayer stabilized by the scaffold protein and by the polymerized lipid,
wherein the polymerized lipid is formed by crosslinking polymerizable lipids within the membrane forming lipid discoidal bilayer in a molar concentration of about 5 to about 40 mol %.

2. The nanolipoprotein particle of claim 1 , wherein the polymerized lipid is in a molar concentration of about 10 to about 40 mol %.

3. The nanolipoprotein particle of claim 1 , wherein a total lipid to scaffold protein molar percent ratio ranges from 20:1 to 240:1.

4. The nanolipoprotein particle of claim 1 , wherein the membrane forming lipid is in amount from 95 to 60% and the polymerized lipid is in an amount from 5 to 40% with respect to a total lipid concentration.

5. The nanolipoprotein particle of claim 1 , wherein the membrane forming lipid and polymerized lipid are in a molar percent ratio membrane forming lipid: polymerized lipids ranging from 95:5 to 60:40.

6. The nanolipoprotein particle of claim 1 , wherein the polymerizable lipids comprise lipids of Formula (I)

wherein
R1 and R2 are independently selected from C7-C29 branched or straight, substituted or unsubstituted aliphatic carbon chain, at least one of R1 and R2 present at least one polymerizable functional group;

in which R 11 , R 12 and R 13 are independently H or a C1-C4 branched or straight aliphatic carbon chain;

R 21 is H, OH, or a carboxy group
m=0-3; and
n and o are independently 0 and 1.

7. The nanolipoprotein particle of claim 6 , wherein R1, and R2 present at least two polymerizable functional groups for polymerization with corresponding functional groups within the membrane lipid bilayer.

8. The nanolipoprotein particle of claim 1 , wherein the polymerizable lipids comprise lipids of Formula (VI)

wherein
R 1 , R 2 are independently a C7-C29 branched or straight, substituted or unsubstituted aliphatic carbon chain;

in which R 11 , R 12 are independently H or a C1-C4 branched or straight aliphatic carbon chain and R3 is a C7-C29 branched or straight, substituted or unsubstituted aliphatic carbon chain,
n and o are independently 0 and 1;
and wherein at least one of R 1 , R 2 and R 3 comprise at least one polymerizable functional group.

9. The nanolipoprotein particle of claim 8 , wherein at R 1 , R 2 and R 3 present at least two polymerizable functional groups for polymerization with corresponding functional groups within the membrane lipid bilayer.

10. The nanolipoprotein particle of claim 1 , wherein the polymerizable lipids comprise lipids of Formula (X)

wherein
R 4 , R 5 and R 6 are independently C7-C29 branched or straight, substituted or unsubstituted aliphatic carbon chain, at least one of R 4 , R 5 and R 6 contains at least one polymerizable functional group, and at least one of R 4 , R 5 and R 6 contains at least one amino nitrogen.

11. The nanolipoprotein particle of claim 10 , wherein at least one of R 2 , R 3 and R 4 contains 1-6 units of an ethyleneoxy group —(CH 2 CH 2 O)—.

12. The nanolipoprotein particle of claim 6 , wherein the at least one polymerizable functional group in the polymerizable lipids of Formula (I) is selected from diacetylene groups, methacrylate groups, acryloyl groups, sorbyl ester groups, diene groups, styrene groups, vinyl groups and isocyano groups.

13. The nanolipoprotein particle of claim 1 , wherein the nanolipoprotein particle further comprises one or more functionalized amphipathic compound.

14. The nanolipoprotein particle of claim 13 , wherein the one or more functionalized amphipathic compounds comprise one or more functionalized membrane forming lipids.

15. The nanolipoprotein particle of claim 14 , wherein the one or more functionalized membrane forming lipids is present in 0.1 mol % to 95 mol % relative to polymerized lipid in the membrane lipid bilayer.

16. The nanolipoprotein particle of claim 1 , further comprising one or more membrane proteins attached to the membrane lipid bilayer through interaction of the target protein hydrophobic region with the membrane lipid bilayer.

17. The nanolipoprotein particle of claim 1 , further comprising one or more functional molecules selected from a functional molecule embedded in the membrane lipid bilayer, a functional molecule conjugated to a lipophilic anchor compound inserted into the membrane lipid bilayer and a functional molecule conjugated through binding of a functional group with a corresponding functional group presented on functionalized membrane forming lipid of the membrane lipid bilayer.

18. The nanolipoprotein particle of claim 17 , wherein the one or more functional molecule comprises drug or cellular targeting compound.

19. The nanolipoprotein particle of claim 18 , wherein the drug or cellular targeting compound are in an amount ranging from 0.1-10 mol %.

20. A nanolipoprotein particle comprising a cross-linked membrane lipid bilayer confined in a discoidal configuration by a scaffold protein, the cross-linked membrane lipid bilayer comprising one or more polymerized lipids and one or more membrane forming lipids.

21. The nanolipoprotein particle of claim 20 , wherein the one or more polymerized lipids comprise lipids of Formula (I)

wherein
R 1 and R 2 are independently selected from C7-C29 branched or straight, substituted or unsubstituted aliphatic carbon chain, at least one of R 1 and R 2 present at least one polymerizable functional group;

in which R 11 , R 12 and R 13 are independently H or a C1-C4 branched or straight aliphatic carbon chain;
R 21 is H, OH, or a carboxy group
m=0-3; and
n and o are independently 0 and;
and wherein the at least one polymerizable functional group binds a corresponding polymerizable functional group in the membrane lipid bilayer providing a polymerized lipid within the membrane lipid bilayer.

22. The nanolipoprotein particle of claim 21 , wherein at least two polymerizable functional group binds a corresponding polymerizable functional group in the membrane lipid bilayer thus providing a polymerized lipid within the membrane lipid bilayer.

23. The nanolipoprotein particle of claim 20 , wherein the one or more polymerized lipids comprise a lipid of Formula (VI)

wherein
R 1 , R 2 are independen

Assignees

• L Livermore Nat Security Llc
• Univ California

Inventors

• Blanchette Craig D
• Fischer Nicholas
• Gilmore Sean Fitzpatrick
• Rasley Amy
• Henderson Paul

Classifications

• A61K49/0086

  Polymersome, i.e. liposome with polymerisable or polymerized bilayered-forming substances · CPC title

• A61K9/1278

  Post-loading, e.g. by ion or pH gradient · CPC title

• A61K47/6915

  the form being a liposome with polymerisable or polymerized bilayer-forming substances, e.g. polymersomes · CPC title

• C07K14/705 Primary

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title

• C07F9/106

  Adducts, complexes, salts of phosphatides · CPC title