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EP-4737436-A1 - CATIONIC LIPID CONTAINING TWO TERTIARY AMINE GROUPS, AND USE THEREOF

EP4737436A1EP 4737436 A1EP4737436 A1EP 4737436A1EP-4737436-A1

Abstract

The present application provides a novel cationic lipid containing two tertiary amine groups, represented by the general formula (1), wherein each symbol is defined as described herein. This cationic lipid containing two tertiary amine groups is pharmaceutically acceptable, biodegradable, or highly biocompatible, offering advantages such as low toxicity, low immunogenicity, and high biocompatibility. One or more biodegradable groups are incorporated between the hydrophilic head and lipophilic tail chains of the cationic lipid containing two tertiary amine groups in the present application, which prevents endosomal accumulation in the LNPs prepared therefrom, enhances the endosomal escape efficiency of LNPs within cells, and promotes the release of drugs into the cytoplasm to exert their effects.

Inventors

  • LIN, SHENG
  • LIN, Minggui
  • WANG, LINLIN
  • WANG, Ailan
  • WEI, GUOHUA
  • ZHU, QI
  • WENG, WENGUI
  • LIU, CHAO

Assignees

  • Xiamen Sinopeg Biotech Co., Ltd

Dates

Publication Date
20260506
Application Date
20240528

Claims (20)

  1. A cationic lipid, wherein the structure is represented by the general formula (1): wherein, R a , R b , R c and R d are each independently a C 1-3 alkyl group; or, R a and R b , together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group; or, R c and R d , together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group; M is O, S, or NH; L d is a linking bond or a divalent linking group; X is N or CR m , and the R m is H or a C 1-12 alkyl group; L A is represented by -B 3 -L 3 -B 1 -L 1 -R 1 , and L B is represented by -B 4 -L 4 -B 2 -L 2 -R 2 ; wherein, L 1 , L 2 , L 3 , and L 4 are each independently a linking bond or a divalent linking group L; B 1 , B 2 , B 3 , and B 4 are each independently a linking bond or a C 1-20 alkylene group; R 1 and R 2 are each independently a C 1-30 hydrocarbon group or a C 1-30 hydrocarbon derivative residue containing 1 to 4 units of -O-; or a pharmaceutically acceptable salt, tautomer, stereoisomer, deuterated derivative, or solvate thereof.
  2. The cationic lipid according to claim 1, wherein L d is selected from the group consisting of a linking bond, -(CH 2 ) t -, -(CH 2 ) t Z-, -(CH 2 ) t Z-(CH 2 ) t Z-, and -(CH 2 ) t Z(CH 2 ) t Z-; wherein, t is independently an integer from 1 to 12 at each occurrence; Z is independently selected from the group consisting of -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -O-, -S-, -C(=O)S-, -SC(=O)-, -NRC(=O)-, -C(=O)NR g , -NR g C(=O)NR g -, -OC(=O)NR g -, -NR g C(=O)O-, -SC(=O)NR g -, and -NR g C(=O)S- at each occurrence; wherein, R g is independently H or a C 1-12 alkyl group at each occurrence; L d is preferably selected from the group consisting of a linking bond, -(CH 2 ) t -, -(CH 2 ) t O-, -(CH 2 ) t C(=O)-, -(CH 2 ) t C(=O)O-, -(CH 2 ) t OC(=O)-, -(CH 2 ) t C(=O)NH-, -(CH 2 ) t NHC(=O)-, -(CH 2 ) t OC(=O)O-, -(CH 2 ) t NHC(=O)O-, -(CH 2 ) t OC(=O)NH-, -(CH 2 ) t NHC(=O)NH-, -(CH 2 ) t O(CH 2 ) t -, -(CH 2 ) t C(=O)(CH 2 ) t -, -(CH 2 ) t C(=O)O(CH 2 ) t -, -(CH 2 ) t OC(=O)(CH 2 ) t -, -(CH 2 ) t C(=O)NH(CH 2 ) t -, -(CH 2 ) t NHC(=O)(CH 2 ) t -, -(CH 2 ) t OC(=O)O(CH 2 ) t -, -(CH 2 ) t NHC(=O)O(CH 2 ) t -, -(CH 2 ) t OC(=O)NH(CH 2 ) t -, and -(CH 2 ) t NHC(=O)NH(CH 2 ) t -; most preferably, L d is a linking bond or -(CH 2 ) t -.
  3. The cationic lipid according to claim 1, wherein L is independently selected from the group consisting of -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -NH-, -O(CR g R g ) s O-, -S-, -C(=O)S-, -SC(=O)-, -NR g C(=O)-, -C(=O)NR g -, -NR g C(=O)NR g -, -OC(=O)NR g -, -NR g C(=O)O-, -SC(=O)NR g -, -NR g C(=O)S-, -C(=S)-, -OC(=S)-, -C(=S)O-, -OC(=S)O-, -NR g C(=S)-, -C(=S)NR g -, -NR g C(=S)NR g -, -OC(=S)NR g -, and -NR g C(=S)O- at each occurrence; wherein, R g is independently a hydrogen atom or a C 1-12 alkyl group at each occurrence, and s is 2, 3 or 4; preferably, L 1 , L 2 , L 3 , and L 4 are selected from any of the following cases: Case (1): L 1 , L 2 , L 3 , and L 4 are each independently L; Case (2): any one of L 1 , L 2 , L 3 , and L 4 is a linking bond, and the other three are each independently L; preferably, L 3 is a linking bond, and L 1 , L 2 , and L 4 are each independently L; Case (3): any two of L 1 , L 2 , L 3 , and L 4 are linking bonds, and the other two are each independently L; preferably, L 3 and L 4 are linking bonds, and L 1 and L 2 are each independently L; Case (4): any three of L 1 , L 2 , L 3 , and L 4 are linking bonds, and the other is L; Case (5): L 1 , L 2 , L 3 , and L 4 are all linking bonds; more preferably, L 1 , L 2 , L 3 , and L 4 are selected from any one of the aforementioned Case (1) to Case (3); most preferably, L 1 , L 2 , L 3 , and L 4 are selected from any of the following cases: Case (a): L 1 , L 2 , L 3 , and L 4 are each independently selected from the group consisting of -OC(=O)-, -C(=O)O-, -C(=O)NH-, -OC(=O)O-, and -NHC(=O)O-; Case (b): L 3 is a linking bond; L 1 , L 2 , and L 4 are each independently selected from the group consisting of -OC(=O)-, -C(=O)O-, -C(=O)NH-, -OC(=O)O-, and -NHC(=O)O-; Case (c): L 3 and L 4 are linking bonds; L 1 and L 2 are each independently selected from the group consisting of -OC(=O)-, -C(=O)O-, -C(=O)NH-, -OC(=O)O-, and -NHC(=O)O-.
  4. The cationic lipid according to claim 1, wherein B 1 , B 2 , B 3 , and B 4 are selected from any of the following cases: Case (1): B 1 , B 2 , B 3 , and B 4 are each independently a C 1-20 alkylene group, more preferably a C 1-10 alkylene group; Case (2): any one of B 1 , B 2 , B 3 , and B 4 is a linking bond, and the other three are C 1-10 alkylene groups; preferably, B 3 is a linking bond, with B 1 , B 2 , and B 4 each independently being a C 1-10 alkylene group; Case (3): any two of B 1 , B 2 , B 3 , and B 4 are linking bonds, and the other two are C 1-10 alkylene groups; preferably, B 3 and B 4 are linking bonds, with B 1 and B 2 each independently being a C 1-10 alkylene group; Case (4): any three of B 1 , B 2 , B 3 , and B 4 are linking bonds, and the other is a C 1-10 alkylene group; `Case (5): B 1 , B 2 , B 3 , and B 4 are all linking bonds; more preferably, B 1 , B 2 , B 3 , and B 4 are selected from any one of the aforementioned Case (1) to Case (3); wherein, the aforementioned C 1-10 alkylene group is preferably selected from the group consisting of a methylene group, an ethylene group, a propylene group, a butylene group, a pentylene group, a hexylene group, a heptylene group, and an octylene group.
  5. The cationic lipid according to claim 1, wherein R 1 and R 2 are selected from any of the following cases: Case (1): both R 1 and R 2 are C 1-30 hydrocarbon groups; Case (2): both R 1 and R 2 are C 1-30 hydrocarbon derivative residues; Case (3): one of R 1 and R 2 is a C 1-30 hydrocarbon group, and the other is a C 1-30 hydrocarbon derivative residue; the C 1-30 hydrocarbon derivative residue is represented by wherein, the tn is independently an integer from 0 to 12 at each occurrence; R e and R f are each independently selected from the group consisting of a C 1-15 alkyl group, a C 2-15 alkenyl group, and a C 2-15 alkynyl group.
  6. The cationic lipid according to claim 5, wherein the C 1-30 hydrocarbon group is a linear C 1-30 hydrocarbon group or a branched C 1-30 hydrocarbon group; the linear C 1-30 hydrocarbon group is selected from the group consisting of a linear C 1-30 alkyl group, a linear C 2-30 alkenyl group, and a linear C 2-30 alkynyl group; preferably, the linear C 1-30 hydrocarbon group is selected from the group consisting of a linear C 1-25 alkyl group, a linear C 2-25 alkenyl group, and a linear C 2-25 alkynyl group; more preferably, the linear C 1-30 hydrocarbon group is a linear C 2-25 alkenyl group; the branched C 1-30 hydrocarbon group is selected from the group consisting of a branched C 1-30 alkyl group, a branched C 2-30 alkenyl group, and a branched C 2-30 alkynyl group, each independently represented by wherein, tm is an integer from 0 to 12; R e and R f are each independently selected from the group consisting of a C 1-15 alkyl group, a C 2-15 alkenyl group, and a C 2-15 alkynyl group; the branched C 1-30 hydrocarbon group is preferably selected from the group consisting of the following structures: is preferably selected from the group consisting of is further preferably selected from the group consisting of the following structures:
  7. The cationic lipid according to claim 1, wherein L A and L B are each independently selected from the group consisting of the following structures: wherein, g is independently an integer from 1 to 10 at each occurrence, more preferably 1, 2, or 3.
  8. The cationic lipid according to claim 1, wherein R a and R b are identical and are both methyl groups or ethyl groups; R c and R d are identical and are both methyl groups or ethyl groups; or, R a and R b , together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group; R c and R d , together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group; the nitrogen-containing heterocyclic group is more preferably, R a , R b , R c , and R d are all methyl groups.
  9. The cationic lipid according to claim 1, wherein the structure of the cationic lipid is represented by the following general formula (1-A) or (1-B): preferably, the structure of the cationic lipid is represented by general formula (2-1) or (2-2): more preferably, the structure of the cationic lipid is selected from the group consisting of the following general formulas: and in the formulas (3-1) to (3-30), B 1 , B 2 , B 3 , and B 4 are not linking bonds; more preferably, B 1 , B 2 , B 3 , and B 4 are each independently a C 1-10 alkylene group at each occurrence.
  10. The cationic lipid according to claim 1, wherein the structure of the cationic lipid is selected from the group consisting of the following structures: and or, the structure of the cationic lipid is selected from the group consisting of the following structures: and
  11. A lipid composition containing a cationic lipid of any one of claims 1 to 10.
  12. The lipid composition according to claim 11, which additionally contains one or more types of lipids selected from the group consisting of phospholipid, steroid lipid, and PEGylated lipid, and is selected from any one of the following cases: Case (1): the lipid composition additionally contains a phospholipid; Case (2): the lipid composition additionally contains a steroid lipid; Case (3): the lipid composition additionally contains a PEGylated lipid; Case (4): the lipid composition additionally contains a phospholipid and a steroid lipid; Case (5): the lipid composition additionally contains a phospholipid and a PEGylated lipid; Case (6): the lipid composition additionally contains a steroid lipid and a PEGylated lipid; Case (7): the lipid composition additionally contains a phospholipid, a steroid lipid, and a PEGylated lipid; more preferably, the lipid composition additionally contains a phospholipid lipid, a steroid lipid, and a PEGylated lipid, simultaneously.
  13. The lipid composition according to claim 12, wherein the phospholipid is selected from the group consisting of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoleoyl-sn-glycero-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-diundecanoyl-sn-glycero-3-phosphocholine, 1-plamitoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1,2-di- O -octadecenyl-sn-glycero-3-phosphocholine, 1-oleoyl-2-cholesterylhemisuccinyl-sn-glycero-3-phosphocholine, 1-hexadecyl-sn-glycero-3-phosphocholine, 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didecosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-diphytanyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didecosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt, dioleoyl phosphatidylserine, dipalmitoyl phosphatidylglycerol, palmitoyloleoyl phosphatidylethanolamine, distearoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, dimyristoleoyl phosphoethanolamine, 1-stearoyl-2-oleoyl-stearoylethanolamine, 1-stearoyl-2-oleoyl-phosphatidylcholine, sphingomyelin, phosphatidylcholine, phosphatidylethnolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoyl phosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, and combinations thereof; or, the steroid lipid is selected from the group consisting of cholesterol, coprostanol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, and combinations thereof; or, the PEGylated lipid is selected from the group consisting of polyethylene glycol-1,2-dimyristoylglycerol, polyethylene glycol-distearoylphosphatidylethanolamine, PEG-cholesterol, polyethylene glycol-diacylglycerol, and polyethylene glycol-dialkyloxypropyl, and is specifically selected from the group consisting of polyethylene glycol 500-dipalmitoylphosphatidylcholine, polyethylene glycol 2000-dipalmitoylphosphatidylcholine, polyethylene glycol 500-distearylphosphatidylethanolamine polyethylene glycol 2000-distearylphosphatidylethanolamine, polyethylene glycol 500-1,2-oleoylphosphatidylethanolamine, polyethylene glycol 2000-1,2-oleoylphosphatidylethanolamine, polyethylene glycol 2000-2,3-dimyristoylglycerol, and combinations thereof; or, the PEGylated lipid is selected from the group consisting of the following structures: and wherein, n 1 is an integer from 25 to 300, and more preferably, n 1 is selected from the group consisting of 44, 45, 46, 47 and 48.
  14. The lipid composition according to any one of claims 11 to 13, which contains 20% to 80% cationic lipid, 5% to 15% phospholipid, 25% to 55% steroid lipid, and 0.5% to 10% PEGylated lipid, wherein the percentages are the molar percentages of each lipid in the total lipids.
  15. The lipid composition according to claim 12, wherein the molar percentage of cationic lipid in the total lipids is 30% to 65%, more preferably 35%, 40%, 45%, 46%, 47%, 48%, 49%, 50%, or 55%; or, the molar percentage of phospholipid in the total lipids is 7.5% to 13%, more preferably 8%, 9%, 10%, 11%, or 12%; or, the molar percentage of steroid lipid in the total lipids is 35% to 50%, more preferably 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50%; or, the molar percentage of PEGylated lipid in the total is 0.5% to 5%, preferably 1% to 3%, and more preferably 1.5%, 1.6%, 1.7%, 1.8%, or 1.9%.
  16. A lipid pharmaceutical composition containing a lipid composition of any one of claims 11 to 15 and a drug, wherein the drug is selected from the group consisting of a nucleic acid drug, a gene vaccine, an antitumor drug, a small molecule drug, a peptide drug, and a protein drug.
  17. The lipid pharmaceutical composition according to claim 16, wherein the nucleic acid drug is selected from the group consisting of RNA, DNA, antisense nucleic acid, plasmid, interfering nucleic acid, aptamer, antagomir, and ribozyme; wherein, the RNA is selected from the group consisting of mRNA, saRNA, circRNA, miRNA, and siRNA; preferably, the nucleic acid drug is selected from the group consisting of DNA, mRNA, miRNA, and siRNA.
  18. A lipid pharmaceutical composition according to any one of claims 16 to 17, wherein the lipid pharmaceutical composition is used as a drug, and is selected from the group consisting of the following drugs: an antineoplastic agent, an antiviral agent, an antifungal agent, and a vaccine.
  19. A formulation of lipid pharmaceutical composition containing a lipid pharmaceutical composition of any one of claims 15 to 17 and a pharmaceutically acceptable diluent or excipient; the diluent or excipient is preferably deionized water, ultrapure water, phosphate buffer, or normal saline, more preferably phosphate buffer or normal saline, and most preferably normal saline.
  20. A liposome or lipid nanoparticle containing a lipid composition of any one of claims 11 to 15.

Description

TECHNICAL FIELD The present application belongs to the field of drug delivery, relating to a cationic lipid in general, in particular to a cationic lipid containing two tertiary amine groups, which can be used as a drug carrier component, and a lipid composition containing the cationic lipid, a lipid pharmaceutical composition, and formulations and applications thereof. BACKGROUND Lipid nanoparticles (LNPs) are spherical particles made of homogeneous lipids, which enter cells through endocytosis and are transported to endosomes. Then, through the endosomal escape process, the contents of the LNP, such as nucleic acids, are released into the cytoplasm. In the field of delivery, LNP has the advantages of easy preparation, modularity, good biocompatibility and strong payload capacity, and is widely used to deliver various drugs, such as small molecule drug, peptide, protein, nucleic acid, etc., in order to improve the stability and solubility of drugs, enhance the delivery effect, achieve directional delivery and reduce the toxic and side effects of drugs. LNP drug delivery systems also have the advantage of being highly tunable in nature, and their stability, uptake, toxicity, side effects, drug release rates, and targeting properties can be adjusted by changing lipid component contents or lipid species, adding targeting functional molecules, and surface modifications. In addition, LNPs support both re-administration and transient administration, and the safety has been validated through the widespread use of COVID-19 vaccines. LNPs typically contain four lipid components, including a phospholipid, a cationic lipid, a cholesterol, and a PEGylated lipid; wherein, the cationic lipid interacts with drug molecules (e.g., negatively charged nucleic acids) through electrostatic interaction; the phospholipid lipid prevents lipid oxidation or link ligands to the surface of lipid nanoparticles. The steroid lipid has strong membrane fusion ability, which promotes the intracellular uptake and cytosolic entry of drug molecules. The PEGylated lipid is located on the surface of lipid nanoparticles, which can improve the hydrophilicity, avoid rapid removal by the immune system, prevent particle aggregation, and increase stability. Despite recent advances in the use of cationic lipid for drug delivery, there is a need for alternative cationic lipids suitable for conventional therapeutic use. In literature CN114751835A, a series of ionizable tertiary amine lipids containing multiple ester bonds were prepared, and the preparation method was complex and cumbersome, requiring at least seven chemical reactions to obtain a cationic lipid with asymmetrical tail chains and two tertiary amine structures. A number of lipids featuring an amino acid core have been disclosed in CN101674853A; wherein, the amino acid-based lipid with a lysine core contains only one tertiary amine structure and two saturated and unsaturated tails, which is not conducive to the formation of conical geometry, incapable of promoting the destabilization of endosomal membranes and the intracytoplasmic release of nucleic acids. The object of the present application is to provide a class of cationic lipids containing two tertiary amine groups with simple preparation methods, low cytotoxicity, and high gene transfection efficiency in vitro, and synthesis methods and application thereof. SUMMARY To solve the problems above, the application provides a novel cationic lipid and preparation methods thereof, a lipid composition containing the cationic lipid, a lipid pharmaceutical composition containing the lipid composition and formulation thereof, and a liposome or a lipid nanoparticle containing the lipid composition; especially, the LNP-nucleic acid pharmaceutical composition containing the cationic lipid and formulation thereof are provided, with the advantages of high delivery efficiency, safety and low toxicity, and high biocompatibility, which can improve the therapeutic and/or preventive effects of drugs. The above-described purposes of this application can be realized via the embodiments below. In one embodiment, provided herein is a cationic lipid: A cationic lipid, wherein the structure is represented by the general formula (1): wherein, Ra, Rb, Rc and Rd are each independently a C1-3 alkyl group; or, Ra and Rb, together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group; or, Rc and Rd, together with the nitrogen atom to which they are connected, form a nitrogen-containing heterocyclic group;M is O, S, or NH;Ld is a linking bond or a divalent linking group;X is N or CRm, and the Rm is H or a C1-12 alkyl group;LA is represented by -B3-L3-B1-L1-R1, and LB is represented by -B4-L4-B2-L2-R2; wherein, L1, L2, L3, and L4 are each independently a linking bond or a divalent linking group L; B1, B2, B3, and B4 are each independently a linking bond or a C1-20 alkylene group; R1 and R2 are each independently a C1-30 hydroc