Lipid-derived neutral nanoparticles

Patent metadata

Field	Value
Publication number	US-10137086-B2
Application number	US-201815878131-A
Country	US
Kind code	B2
Filing date	Jan 23, 2018
Priority date	Mar 29, 2012
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

Disclosed herein are novel lipids and liposomal compositions prepared using such compounds and related methods of neutralizing or otherwise modifying such liposomal compositions. The lipids described herein are useful for example, as liposomal vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and the subsequent transfection of such target cells. In certain embodiments, one or more of the compounds that comprise the liposomal delivery vehicle may be neutralized or further modified such that the properties of the liposomal delivery vehicle are modified.

First claim

We claim:

1. A composition comprising a lipid nanoparticle having an overall neutral surface charge and encapsulating negatively charged mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein, wherein said lipid nanoparticle is made by a method comprising the steps of:
(a) encapsulating said negatively charged mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein within a lipid nanoparticle, wherein the lipid nanoparticle comprises:
a cationic lipid, the cationic lipid comprising a polar head-group bound to a lipophilic tail-group via a linker group, the polar head-group being exposed on the surface of the lipid nanoparticle, and the linker group being susceptible to chemical or enzymatic cleavage;
a PEG-modified lipid;
one or more non-cationic lipids that is zwitterionic or anionic; and
one or more non-cationic lipids that is neutral;
and
(b) exposing the lipid nanoparticle to chemical or enzymatic cleavage thereby releasing the polar head-group from the surface of the lipid nanoparticle to provide an overall neutral surface charge to the lipid nanoparticle.

2. The composition of claim 1 , wherein the encapsulation efficiency of step (a) is at least 75%.

3. The composition of claim 2 , wherein the encapsulation efficiency is at least 90%.

4. The composition of claim 1 , wherein following step (b) the lipid nanoparticle has a surface charge or zeta potential of about −2.5 to about +2.5 mV.

5. The composition of claim 1 , wherein the cationic lipid with the releasable polar head group is represented by the structural formula:

or a pharmaceutically acceptable salt thereof,
wherein:
R 1 is selected from the group consisting of imidazole; guanidinium; imine; enamine; amino; an alkyl amino optionally-substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide; or a pyridyl optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide;
R 2 is selected from the group consisting of alkyl, alkenyl, acyl, pyridyl,

each optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, nitro, or amide;
R 3 and R 4 are each independently selected from the group consisting of C 6 -C 20 alkyl, C 6 -C 20 alkenyl, and C 6 -C 20 acyl, each optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide; and
n is zero or any positive integer;
and
wherein the modulating step comprises exposing the lipid nanoparticle to a reducing agent thereby cleaving the linker group and releasing the polar head-group from the lipophilic tail-group.

6. The composition of claim 1 , wherein the cationic lipid is selected from the group consisting of:

7. The composition of claim 1 , wherein the composition comprises the following structure after the modulating step:

or a pharmaceutically acceptable salt thereof,
wherein:
R 2 is selected from the group consisting of alkyl, alkenyl, acyl, pyridyl,

each optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, nitro, or amide;
R 3 and R 4 are each independently selected from the group consisting of C 5 -C 20 alkyl, C 6 -C 20 alkenyl, and C 6 -C 20 acyl, each optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide; and
n is zero or any positive integer;
and
wherein the modulating step comprises exposing the lipid nanoparticle to a reducing agent thereby cleaving the linker group and releasing the polar head-group from the lipophilic tail-group.

8. The composition of claim 1 , wherein said one or more non-cationic lipids that is zwitterionic or anionic is: distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), DLPE (1,2-dilauroyl-sn-glycero-3-phosphoethanolamine), DPPS (1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), a sphingomyelin, or a mixture thereof.

9. The composition of claim 1 , wherein said one or more non-cationic lipids that is neutral comprises cholesterol.

10. A pharmaceutical composition comprising a lipid nanoparticle having an overall neutral surface charge and encapsulating negatively charged mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein, wherein said lipid nanoparticle is made by a method comprising the steps of:
(a) encapsulating said negatively charged mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein within a lipid nanoparticle, wherein the lipid nanoparticle comprises:
a cationic lipid, the cationic lipid comprising a polar head-group bound to a lipophilic tail-group via a linker group, the polar head-group being exposed on the surface of the lipid nanoparticle, and the linker group being susceptible to chemical or enzymatic cleavage;
a PEG-modified lipid;
one or more non-cationic lipids that is zwitterionic or anionic; and
one or more non-cationic lipids that is neutral; and
(b) exposing the surface of the lipid nanoparticle to chemical or enzymatic cleavage thereby releasing the polar head-group from the surface of the lipid nanoparticle to provide an overall neutral surface to the lipid nanoparticle.

11. The pharmaceutical composition of claim 10 , wherein the encapsulation efficiency of step (a) is at least 75%.

12. The pharmaceutical composition of claim 11 , wherein the encapsulation efficiency is at least 90%.

13. The pharmaceutical composition of claim 10 , wherein following step (b) the lipid nanoparticle has a surface charge or zeta potential of about −2.5 to about +2.5 mV.

14. The pharmaceutical composition of claim 10 , wherein the cationic lipid with the releasable polar head group is represented by the structural formula:

or a pharmaceutically acceptable salt thereof,
wherein:
R 1 is selected from the group consisting of imidazole; guanidinium; imine; enamine; amino; an alkyl amino optionally-substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide; or a pyridyl optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide;
R 2 is selected from the group consisting of alkyl, alkenyl, acyl, pyridyl,

each optionally substituted with alkyl, halo, alkoxy, hydroxyl, amino, aryl, ether, ester, nitro, or amide;

Assignees

• Translate Bio Inc

Inventors

• Derosa Frank
• Guild Braydon Charles
• Heartlein Michael

Classifications

• A61K9/1271

  Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) · CPC title

• A61K9/1277 Primary

  Preparation processes; Proliposomes · CPC title

• A61K38/44

  Oxidoreductases (1) · CPC title

• C12N2320/32

  Special delivery means, e.g. tissue-specific · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title