EP-4734959-A1 - USE OF GLYCERIDES FOR LNP FORMULATIONS

Abstract

Provided are lipid nanoparticles for delivering nucleic acids molecules such as mRNA. Also provided are methods of making and using thereof.

Inventors

• DEROSA, FRANK
• KARVE, Shrirang
• KHADSE, Nikita
• SARODE, ASHISH

Assignees

• Sanofi Pasteur Inc.

Dates

Publication Date

20260506

Application Date

20240628

Claims (1)

1. 755643: SA9-383PC CLAIMS 1. A composition comprising a lipid nanoparticle (LNP), wherein the LNP comprises: (I) an ionizable lipid; (II) a glyceride or an acylglycol; and (III) one or more lipids selected from the group consisting of: (a) a structural lipid; (b) a helper lipid; and (c) a stealth lipid. 2. The composition of claim 1, wherein the LNP comprises: (I) an ionizable lipid; (II) a glyceride or an acylglycol; (III) a structural lipid; (IV) a helper lipid; and (V) a stealth lipid. 3. The composition of claim 1 or claim 2, wherein the glyceride or acylglycol is a monoglyceride, a diglyceride, a triglyceride, or a diacylglycol. 4. The composition of claim 1 or claim 2, wherein the glyceride or acylglycol has a structure according to Formula I or Formula II: (I) (II) wherein: R G1 , R G2 , and R G3 are each independently H, -C (1-25) alkyl, -C (1-25) alkenyl, -C(O)C (1- 25) alkyl, or -C(O)C (1-25) alkenyl, wherein the -C (1-25) alkyl, -C (1-25) alkenyl, -C(O)C (1-25) alkyl, and -C(O)C (1-25) alkenyl are optionally substituted with one to three groups independently selected from -OC(O)C (1-25) alkyl, -OC(O)C (1-25) alkenyl, -C(O)OC (1-25) alkyl, -C(O)OC (1-25) alkenyl, - OC (1-25) alkyl, -OC (1-25) alkenyl, -C(O)C (1-25) alkyl, and -C(O)C (1-25) alkenyl; provided that no more than two of R G1 , R G2 and R G3 are H; R G4 is -C (1-25) alkyl, -C (1-25) alkenyl, -C(O)C (1-25) alkyl, or -C(O)C (1-25) alkenyl, each of which is optionally substituted with one to three groups independently selected from - 755643: SA9-383PC OC(O)C (1-25) alkyl, -OC(O)C (1-25) alkenyl, -C(O)OC (1-25) alkyl, -C(O)OC (1-25) alkenyl, -OC (1- 25) alkyl, -OC (1-25) alkenyl, -C(O)C (1-25) alkyl, and -C(O)C (1-25) alkenyl; R G5 is H, -C (1-25) alkyl, -C (1-25) alkenyl, -C(O)C (1-25) alkyl, or -C(O)C (1-25) alkenyl, wherein the -C (1-25) alkyl, -C (1-25) alkenyl, -C(O)C (1-25) alkyl, and -C(O)C (1-25) alkenyl are optionally substituted with one to three groups independently selected from -OC(O)C (1- 25) alkyl, -OC(O)C (1-25) alkenyl, -C(O)OC (1-25) alkyl, -C(O)OC (1-25) alkenyl, -OC (1-25) alkyl, -OC (1- 25) alkenyl, -C(O)C (1-25) alkyl, and -C(O)C (1-25) alkenyl. 5. The composition of claim any one of claims 1-4, wherein the glyceride or acylglycol is hydrolyzable by lipase. 6. The composition of any one of claims 1-3, wherein the glyceride or acylglycol is selected from the group consisting of: (S) 1-C16 Ether MG O OH HO H OH Monoolein O OH O C 18(plasm) MG O H O H OH OH Monolinolein O OH O O 08:0 DG O OH O H O O 10:0 DG O OH O H O O 12:0 DG O OH O H O O 14:0 DG O OH O H O O 15:0-18:1 DG O OH O O O 16:0 Ethylene O Glycol O O O 16:0 DG O OH O H O 755643: SA9-383PC O :0-18:1 DG O OH O H O O 18:0 DG O OH O H O O O Diolein OH O O O :0-16:0 DG O OH O H O O :0-18:2 DG O OH O H O O :0-20:4 DG O OH O H O O :0-22:6 DG O OH O H O O :1 Ethylene O Glycol O O O 18:1 DG O OH O H O O 8:1-2:0 DG O OH O H O O 18:2 DG O OH O H O O O Dilinolein OH O O O O O Tributyrin O O H O O O O Tricaproin O O H O 755643: SA9-383PC O O O Trioctanoin O O H O O O O Tricaprin O O H O O O O Trilaurin O O H O O O O Trimyristin O O H O O O 5:0-18:1-15:0 TG O O O O 6:0-(12-PAHSA)- 18:1 TG O O O Tristearin O O H O O O O Triolein O O H O O O O Trilinolein O O H O Glyceryl O O O trinonadecanoate O O H O O O O Triarachidin O O H O 755643: SA9-383PC O O O Tripalmitin O O H O or a combination thereof. 7. The composition of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein: the ionizable lipid has a structure according to Formula CAT-I: (CAT-I), wherein: p is an integer of between 1 and 9, inclusive; each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl; each instance of L is independently an optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene, or combination thereof; each instance of R 6 and R 7 is independently a group of formula (i), (ii), or (iii); Formulae (i), (ii), and (iii) are: 755643: SA9-383PC (iii), wherein: each instance of R′ is independently hydrogen or optionally substituted alkyl; X is O, S, or NR X , wherein R X is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; Y is O, S, or NR Y , wherein R Y is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; R P is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to a sulfur atom, or a nitrogen protecting group when attached to a nitrogen atom; and R L is optionally substituted C 1-50 alkyl, optionally substituted C 2-50 alkenyl, optionally substituted C 2-50 alkynyl, optionally substituted heteroC 1-50 alkyl, optionally substituted heteroC 2-50 alkenyl, optionally substituted heteroC 2-50 alkynyl, or a polymer; or the ionizable lipid has a structure according to Formula CAT-II: side of each depicted structure is bound to the -(CH 2 )a-; 755643: SA9-383PC O O Z 1 is selected from , wherein the right hand side of each depicted structure is bound to the -(CH 2 )a-; R 1A and R 1B are each independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted acyl, and -W 1 -X 1 -Y 1 ; each W 1 is independently selected from optionally substituted alkyl and optionally substituted alkenyl; each X 1 is independently selected from -*O-(C=O)-optionally substituted alkyl, - (*C=O)-O-optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and - (*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W 1 , each Y 1 is independently selected from hydrogen, -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O-optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to X 1 ; b is 1, 2, 3, 4, or 5; and each a is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. 8. The composition of any one of claims 1-6, wherein the ionizable lipid is selected from 755643: SA9-383PC O O N HO N HO N OH N S S OH OH HO O O N N O O N HO OH OH HO O OH O N O O N OH OH OH OH N O N O N S N S OH HO C 10 H 21 OH C 10 H 21 OH O N O N N N O O HO C10 H 21 HO C 10 H 21 O O O O O O O O O O N O O O O 9. The composition of any one of claims 1-8, wherein: the structural lipid is a sterol, for example cholesterol; the helper lipid is 1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE); 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC); 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS); 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (DEPE); and 1,2-dioleoyl-sn- 755643: SA9-383PC glycero-3-phosphocholine (DPOC), dipalmitoylphosphatidylcholine (DPPC), 1,2-dilauroyl- sn-glycero-3-phosphocholine (DLPC), 1,2-Distearoylphosphatidylethanolamine (DSPE), or 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE); and the stealth lipid is a polyethylene glycol-conjugated (PEGylated) lipid selected from the group consisting of 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG- PEG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol (DSPE-PEG), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol (DLPE-PEG), and 1,2- distearoyl-rac-glycero-polyethelene glycol (DSG-PEG). 10. The composition of any one of claims 1-9, wherein the LNP comprises: the ionizable lipid at a molar ratio between 35% and 55%, the structural lipid at a molar ratio between 20% and 35%, the stealth lipid at a molar ratio between 0.25% and 2.75%, and the helper lipid and the glyceride or acylglycol at a combined molar ratio of between 10% and 35%. 11. The composition of claim 1, wherein LNP comprises: GL-HEPES-E3-E12-DS-4-E10 at a molar ratio of about 40%, cholesterol at a molar ratio of about 28.5%, DMG-PEG2000 at a molar ratio of about 1.5%, DOPE at a molar ratio of about 25%, and trimyristin at a molar ratio of about 5%. 12. The composition of any one of claims 1-11, further comprising a nucleic acid molecule, wherein the nucleic acid molecule is encapsulated in the LNP, and wherein the nucleic acid molecule is an mRNA molecule. 13. The composition of claim 12, wherein the mRNA molecule encodes an antigen, optionally a viral antigen or a bacterial antigen. 14. The composition of claim 12 or 13, wherein the composition comprises two or more LNPs, wherein each LNP encapsulates an mRNA encoding a different antigen, optionally wherein the different antigens are from the same pathogen or from different pathogens. 755643: SA9-383PC 15. Use of the composition of any one of claims 12-14 for the manufacture of a medicament for eliciting an immune response in a subject. 16. Use of the composition of any one of claims 12-14 for the manufacture of a medicament for preventing an infection or reducing one or more symptoms of an infection. 17. A method of eliciting an immune response in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of the composition of any one of claims 12-14. 18. A method of preventing an infection or reducing one or more symptoms of an infection in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of the composition of any one of claims 12-14.

Description

755643: SA9-383PC Use of Glycerides for LNP Formulations CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to European Patent Application No.23306047.4, filed June 28, 2023, the disclosure of which is incorporated herein by reference in its entirety. BACKGROUND Effective targeted delivery of biologically active substances such as nucleic acid molecules (e.g., mRNA) represents a continuing medical challenge. In particular, the delivery of nucleic acids to cells is made difficult by their low in vivo stability, propensity toward rapid degradation, and low cell permeability. Thus, there exists a need to develop methods and compositions to facilitate the delivery of therapeutic and/or prophylactics such as nucleic acids to cells. Lipid-containing nanoparticle compositions have proven effective as transport vehicles into cells and/or intracellular compartments for biologically active substances such as small molecule drugs, proteins, and nucleic acids. Such compositions generally include one or more ionizable (e.g., cationic) lipids, phospholipids including polyunsaturated lipids, cholesterol-based lipids, and/or lipids containing polyethylene glycol (PEGylated lipids). Though a variety of such lipid-containing nanoparticle compositions have been demonstrated, there remains a need for lipid nanoparticle formulations having improved efficacy. SUMMARY The present disclosure provides, inter alia, a composition comprising a lipid nanoparticle (LNP), wherein the LNP comprises: (I) an ionizable lipid; (II) a glyceride or an acylglycol; and (III) one or more lipids selected from the group consisting of: (a) a structural lipid; (b) a helper lipid; and (c) a stealth lipid. In some embodiments, the LNP comprises: (I) an ionizable lipid; (II) a glyceride or an acylglycol; (III) a structural lipid; (IV) a helper lipid; and (V) a stealth lipid. In some embodiments, the LNP comprises: (I) an ionizable lipid; (II) a glyceride or an acylglycol having a structure according to Formula I or Formula II, as defined herein; (III) a structural lipid; (IV) a helper lipid; and (V) a stealth lipid. The present disclosure further provides a composition comprising a lipid nanoparticle (LNP), wherein the LNP comprises: (I) an ionizable lipid having a structure according to 755643: SA9-383PC Formula CAT-I or CAT-II, as defined herein; (II) a glyceride or an acylglycol; (III) a structural lipid, (IV) a stealth lipid; and (V) a helper lipid. In some embodiments, the LNP comprises: (I) an ionizable lipid having a structure according to Formula CAT-I or CAT-II, as defined herein; (II) a glyceride or an acylglycol having a structure according to Formula I or II, as defined herein; (III) a structural lipid, (IV) a stealth lipid; and (V) a helper lipid. The present disclosure further provides an LNP as described herein, further comprising a nucleic acid molecule, wherein the nucleic acid molecule is encapsulated in the LNP. In some embodiments, the nucleic acid molecule is an mRNA molecule. The present disclosure further provides a method of preventing an infection or reducing one or more symptoms of an infection, comprising administering to the subject, optionally intramuscularly, intranasally, intravenously, subcutaneously, or intradermally, a prophylactically effective amount of a composition described herein. The present disclosure further provides the use of a composition described herein for the manufacture of a medicament for use in treating a subject in need thereof. The present disclosure further provides a kit comprising a container comprising a single-use or multi-use dosage of a composition described herein, optionally wherein the container is a vial or a pre-filled syringe or injector. DETAILED DESCRIPTION The present disclosure provides lipid nanoparticle (LNP) formulations for delivering cargo, such as a nucleic acid molecule (e.g., mRNA), to a target cell. In particular, the LNPs of the present disclosure comprise an ionizable lipid, a glyceride or an acylglycol, and at least one of a structural lipid, a helper lipid, and a stealth lipid (e.g., PEGylated). In some embodiments, the LNPs comprise an ionizable lipid, a glyceride or an acylglycol, a structural lipid, a helper lipid, and a stealth lipid. It has been discovered that addition of a glyceride or an acylglycol to LNP formulations can improve the delivery efficiency of the mRNA, thereby increasing the expression of the protein encoded by the nucleic acid molecule when compared to LNP formulations without a glyceride or an acylglycol. For example, LNP formulations of the present disclosure comprising hEPO mRNA were found to significantly improve protein expression over control formulations. 755643: SA9-383PC Definitions Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. As used in the specification and in the claims, the