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EP-4737504-A2 - CATIONIC LIPID, LIPOSOME CONTAINING CATIONIC LIPID, AND NUCLEIC-ACID PHARMACEUTICAL COMPOSITION CONTAINING LIPOSOME AND FORMULATION AND APPLICATION THEREOF

EP4737504A2EP 4737504 A2EP4737504 A2EP 4737504A2EP-4737504-A2

Abstract

The present invention provides a novel cationic lipid having a structure as represented by general formula (1) and specifically relates to a nitrogen-branched cationic lipid, a liposome containing the cationic lipid, a nucleic-acid pharmaceutical composition containing the liposome, a preparation method and application thereof. The definition of each symbol in the formula (1) is as defined in the specification. The cationic liposome containing the cationic lipid as represented by formula (1) can improve the loading rate and transport efficiency of nucleic-acid drugs. The formulation of the cationic liposome nucleic-acid pharmaceutical composition of the present invention has good biocompatibility and higher gene transfection efficiency, and can improve the treatment and/or prevention effects of nucleic-acid drugs.

Inventors

  • WENG, WENGUI
  • LIU, CHAO
  • WANG, Ailan
  • LIN, SHENG

Assignees

  • Xiamen Sinopeg Biotech Co., Ltd.

Dates

Publication Date
20260506
Application Date
20220401

Claims (13)

  1. A cationic lipid, wherein, the structure is represented by the general formula (1): wherein, N is the nitrogen-atom branching center; wherein, one of L 1 and L 2 is -OC(=O)-, -C(=O)O-, or -OC(=O)O-, and the other is - C(=O)O-, -OC(=O)O-, or -C(=O)-; or both L 1 and L 2 are -OC(=O)-; wherein, L 3 is a linking bond; wherein, B 1 and B 2 are each independently a linking bond or a C 1-30 alkylene group; wherein, one of R 1 and R 2 is a C 5-30 aliphatic group or and the other is wherein, t is an integer from 0 to 12; wherein, t 1 and t 2 are each independently an integer from 0 to 5; wherein, t 3 and t 4 are 1; wherein, 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; wherein, R 3 is selected from the group consisting of a hydrogen atom, -R d , -C(=O)R d , and -C(=O)OR d ; wherein, R d is, at each occurrence, independently a C 1-12 alkyl group; wherein, A is -CH 2 CH 2 O-, and n is an integer from 2 to 6.
  2. The cationic lipid according to claim 1, wherein, said B 1 and B 2 are each independently a linking bond or a C 1-20 alkylene group; more preferably, one of B 1 and B 2 is a linking bond, and the other is a C 1-20 alkylene group; more preferably, B 1 and B 2 are each independently 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, an octylene group, a nonylene group, a decylene group, an undecylene group, a dodecylene group, a tridecylene group, a tetradecylene group, a pentadecylene group, a hexadecylene group, a heptadecylene group, an octadecylene group, a nonadecylene group, and an eicosylene group.
  3. The cationic lipid according to claim 1, wherein, said L 1 and L 2 are selected from one of the following situations: (1) L 1 and L 2 are both -C(=O)O-, or both -OC(=O)O-; (2) one of L 1 and L 2 is selected from the group consisting of -OC(=O)-, -C(=O)O-, and - OC(=O)O-, and the other is -C(=O)-.
  4. The cationic lipid according to claim 1, wherein, said R 1 and R 2 are each independentlya nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl group, a heptadecyl group, an octadecyl group, a nonadecyl group, an eicosyl group, a heneicosyl group, a docosyl group, a tricosyl group, a lignoceroylv group, a ( Z )-tridec-8-enyl group, a ( Z )-tetradec-9-enyl group, a ( Z )-pentadec-8-enyl group, a ( Z )-hexadec-9-enyl group, a ( Z )-heptadec-5-enyl group, a ( Z )-heptadec-8-enyl group, an ( E )-heptadec-8-enyl group, a ( Z )-heptadec-10-enyl group, an (8 Z , 11 Z )-heptadec-8,11-dienyl group, a ( Z )-octodec-6-enyl group, a ( Z )-octodec-9-enyl group, an ( E )-octodec-9-enyl group, a ( Z )-octodec-11-enyl group, a (9 Z , 12 Z )-octodec-9,12-dienyl group, a (9 Z , 12 Z , 15 Z )-octodec-9,12,15-trienyl group, an (8 Z , 11 Z, 14 Z )-octodec-8,11,14-trienyl group, a ( Z )-eicos-11-enyl group, an (11 Z , 1 4 Z )-eicos-11,14-dienyl group, a ( Z )-nonadec-10-enyl group, a (10 Z , 13 Z )-nonadec-10,13-dienyl group, a 2,6,10-trimethylundec-1,5,9-trienyl group, a 3,7,11-trimethyldodec-2,6,10-trienyl group, or a 3,7,11,15-tetramethylhexadec-2-enyltridecyl group; or said R 1 and R 2 are each independently represented as or wherein R e and R f are each independently selected from the group consisting of a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a vinyl group, a propenyl group, an allyl group, a butenyl group, an allyl carbinyl group, a pentenyl group, a neopentyl group, a hexenyl group, a neohexenyl group, a heptenyl group, a neoheptenyl group, an octenyl group, a neooctenyl group, a nonenyl group, a neononenyl group, a decenoyl group, a neodecenoyl group, an ethynyl group, a propynyl group, a propargyl group, a butynyl group, a butynediyl group, a pentynyl group, a neopentyl group, a hexynyl group, a neohexyl group, a heptynyl group, a neoheptyl group, an octynyl group, a neooctyl group, a nonynyl group, a neononyl group, a decynylgroup, and a neodecyl group; more preferably R e and R f are each independently selected from the group consisting of a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, and a decyl group; and further more R 1 and R 2 are each independently selected from the group consisting of the following structures: and
  5. The cationic lipid according to claim 1, wherein, the structure of said cationic lipid is selected from the group consisting of the following formulas:
  6. The cationic lipid according to claim 1, wherein, the structure of said cationic lipid includes but is not limited to the following structures: and
  7. A cationic liposome, wherein, the cationic liposome contains any foregoing cationic lipid according to any one of claims 1-6.
  8. The cationic liposome according to claim 7, wherein, the cationic liposome contains one or more lipids selected from the group consisting of neutral lipids, steroid lipids, and PEGylated lipids, more preferably contains neutral lipids, steroid lipids, and PEGylated lipids simultaneously; wherein, said neutral lipid is preferably phospholipid; more preferably wherein, said neutral lipid is selected from the group consisting of 1,2-dilinoleoyl-sn-glycero-3-phosphocholines, 1,2-dimyristoleoyl-sn-glycero-3-phosphocholines, 1,2-dioleoyl-sn-glycero-3-phosphocholines, 1,2-dipalmitoyl-sn-glycero-3-phosphocholines, 1,2-distearoyl-sn-glycero-3-phosphatidylcholines, 1,2-diundecanoyl-sn-glycero-3-phosphatidylcholines, 1-plamitoyl-2-oleoyl-sn-glycero-3-phosphocholines, 1,2-di- O -octadecenyl-sn-glycero-3-phosphatidylcholines, 1-oleoyl-2-cholesterylhemisuccinyl-sn-glycero-3-phosphocholines, 1- O -hexadecyl-sn-glycero-3-phosphatidylcholines, 1,2-dilinolenoyl-sn-glycero-3-phosphatidylcholines, 1,2-diarachidonoyl-sn-glycero-3-phosphatidylcholines, 1,2-didecosahexaenoyl-sn-glycero-3-phosphocholines, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamines, 1,2-diphytanyl-sn-glycero-3-phosphoethanolamines, 1,2-distearoyl-sn-glycero-3-phosphoethanolamines, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamines, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamines, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamines, 1,2-didecosahexaenoyl-sn-glycero-3-phosphoethanolamines, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salts, dioleoyl phosphatidylserines, dipalmitoylphosphatidylglycerols, palmitoyloleoyl phosphatidylethanolamines, distearoyl phosphatidylethanolamines, dipalmitoyl phosphatidylethanolamines, dimyristoleoyl phosphoethanolamines, 1-stearoyl-2-oleoyl- stearoylethanolamines, 1-stearoyl-2-oleoyl-phosphatidylcholines, sphingomyelins, phosphatidylcholines, phosphatidylethnolamines, phosphatidylserines, phosphatidylinositols, phosphatidic acids, palmitoyloleoyl phosphatidylcholines, lysophosphatidylcholines, lysophosphatidylethanolamines, and combinations thereof; pwherein, said steroid lipid is selected from the group consisting of cholesterols, coprostanols, sitosterols, ergosterols, campesterols, stigmasterols, brassicasterol tomatidines, ursolic acids, α-tocopherols, and any mixture thereof; wherein, said PEGylated lipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)], PEG-cholesterol, PEG-diacylglycamide, and PEG-dialkyloxypropyl, specifically includes PEG500-dipalmitoylphosphatidylcholine, PEG2000-dipalmitoylphosphatidylcholine, PEG500-stearylphosphatidylethanolamine, PEG2000-distearylphosphatidylethanolamine, PEG500-1,2-dioleoylphosphatidylethanolamine, PEG2000-1,2-dioleoylphosphatidylethanolamine, and PEG2000-2,3-distearoylglycerol; wherein, the structure of said PEGylated lipid is represented by the general formula (2): or pharmaceutically acceptable salts, tautomers, and stereoisomers thereof; wherein, L 7 and L 8 are each independently a linking bond or a divalent linking group, said divalent linking group is selected from the group consisting of -OC(=O)-, -C(=O)O-, - OC(=O)O-, -C(=O)-, -O-, -S-, -C(=O)S-, -SC(=O)-, -NR c C(=O)-, -C(=O)NR c -, - NR c C(=O)NR c -, -OC(=O)NR c -, -NR c C(=O)O-, -SC(=O)NR c -, and -NR c C(=O)S-; wherein, R c is, at each occurrence, independently a hydrogen atom or a C 1-12 alkyl group; wherein, L 3 is selected from the group consisting of a linking bond, -L 4 -, -Z-L 4 -Z-, -L 4 -Z-L 5 -, -Z-L 4 -Z-L 5 -, and -L 4 -Z-L 5 -Z-; said L 4 and L 5 are carbon-chain linking groups, which are each independently represented by -(CR a R b ) t -(CR a R b ) o -(CR a R b ) p -; wherein, t, o, and p are each independently an integer from 0 to 12, not being 0 simultaneously; wherein, R a and R b are, at each occurrence, independently a hydrogen atom or an alkyl group; wherein, said Z is, at each occurrence, independently selected from the group consisting of -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -O-, -S-, -C(=O)S-, -SC(=O)-, -NR c C(=O)-, -C(=O)NR c -, -NR c C(=O)NR c -, - OC(=O)NR c -, -NR c C(=O)O-, -SC(=O)NR c -, -NR c C(=O)S-, and wherein, R c is, at each occurrence, independently a hydrogen atom or a C 1-12 alkyl group; wherein, B 3 and B 4 are each independently a linking bond or a C 1-12 alkylene group; wherein, R 1 and R 2 are each independently a C 1-30 aliphatic group; wherein, R is selected from the group consisting of a hydrogen atom, an alkyl group, an alkoxy group, -C(=O)R d , -C(=O)OR d , -OC(=O)R d , and -OC(=O)OR d ; wherein, R d is a C 1-12 alkyl group; wherein, A is -(CR a R b ) s O- or -O(CR a R b ) s -; wherein, s is 2, 3 or 4; wherein, R a and R b are each independently a hydrogen atom or a C 1-12 alkyl group; wherein, n 1 is an integer from 20 to 250; wherein, said alkyl group, alkylene group, alkoxy group, and aliphatic group are each independently substituted or unsubstituted; orwherein, the structure of said PEGylated lipid is selected from the following structures: and
  9. The cationic liposome according to claim 8, wherein, cationic liposome contains 20-80% cationic lipids, 5-15% neutral lipids, 25-55% steroid lipids, and 0.5-10% PEGylated lipids , said percentage being the molar percentage of each type of lipid relative to the total lipids in a solution containing a solvent.
  10. The cationic liposome according to any one of claims 8-9, wherein, the molar percentage of cationic lipids relative to the total lipids in a solution containing a solvent is 30% to 65%, preferably about 35%, 40%, 45%, 46%, 47%, 48%, 49%, 50%, or 55%; or wherein, the molar percentage of neutral lipids relative to the total lipids in a solution containing a solvent is 7.5% to 13%, preferably about 8%, 9%, 10%, 11%, or 12%; or wherein, the molar percentage of steroid lipids relative to the total lipids in a solution containing a solvent is 35% to 50%, preferably about 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50%; or wherein, the molar percentage of PEGylated lipids relative to the total lipids in a solution containing a solvent is 0.5% to 5%, preferably 1% to 3%, and more preferably about 1.5%, 1.6%, 1.7%, 1.8%, or 1.9%.
  11. A cationic liposome-nucleic acid pharmaceutical composition, wherein, the cationic liposome-nucleic acid pharmaceutical composition contains the cationic liposome according to any one of claims 8-10 and nucleic acid drug; wherein, said nucleic acid drug is selected from the group consisting of DNA, antisense nucleic acid, plasmid, mRNA, interfering nucleic acid, aptamer, antagomir, miRNA, ribozyme, and siRNA, preferably DNA, mRNA, miRNA, or siRNA.
  12. The cationic liposome-nucleic acid pharmaceutical composition according to claim 11 for use as a medicament in the treatment of a disease selected from cancer and malignant tumor, infection, a viral disease, a fungal disease, or for use in the prevention of a disease wherein the composition is a vaccine.
  13. A formulation of cationic liposome-nucleic acid pharmaceutical composition, wherein, the formulation of cationic liposome-nucleic acid pharmaceutical composition contains the cationic liposome-nucleic acid pharmaceutical composition according to any one of claims 11-12 and pharmaceutically acceptable diluents or excipients; wherein, said diluent or excipient is preferably deionized water, ultrapure water, phosphate buffer, or physiological saline, more preferably phosphate buffer or physiological saline, and most preferably physiological saline.

Description

TECHNICAL FIELD The present invention relates to the field of drug delivery, specifically to a cationic lipid as pharmaceutical carrier, in particular to a cationic lipid branched via nitrogen, liposomes containing the lipid, nucleic acid pharmaceutical compositions containing the liposome, formulations of the compositions and application thereof. BACKGROUND OF THE INVENTION Liposome is a microvesicle formed by encapsulating drugs in the lipid bilayer. Liposomal nanoparticle contains liposomes and nucleic acid drugs, which is similar in structure to biofilm, and is a biocompatible and non-toxic nanomaterial. Liposome can encapsulate water-soluble and lipo-soluble drugs, with advantages such as reducing drug dose, sustained release, targeted drug release and protecting encapsulated nucleic acids from degradation and clearance in serum. In addition, nanoliposome is also an excellent antigen carrier, which can not only encapsulate a series of antigens with different physicochemical properties and immune adjuvants, protect protein polypeptide antigens from degradation, but also promote the phagocytosis and presentation of antigen-presenting cells to antigen, thus enhancing the specific immune response of the body. Therefore, liposomal nanoparticles are widely used in the field of drug delivery. Based on the aforementioned advantages, liposomal nanoparticle, as a new type of vaccine vector, has attracted wide attention. Liposomal nanoparticles are gradually used in the development of antiviral vaccines, antibacterial vaccines, antiparasitic vaccines, antitumor vaccines, and so on. Neutral liposomes and cationic liposomes with the surface carrying positive charges are the most commonly used nano-sized vaccine vectors. Cationic liposomes are not only excellent protein/peptide antigen carriers, but also new immunological adjuvants, which can directly activate antigen-presenting cells and enhance the immune response induced by vaccines. Therefore, cationic liposomes are widely used for encapsulating and transporting nucleic acid drugs in the field of vaccine. The process of encapsulating and transporting nucleic acid drugs by cationic liposomes is as follows: (i) cationic liposome-nucleic acid drug complex forms via electrostatic interaction between positively charged cationic lipids and negatively charged nucleic acids; (ii) electrostatic interaction between the positively charged liposome and the negatively charged cell membrane promotes the adsorption of cationic liposome-nucleic acid drug complexes to the cell surface, followed by endocytosis and endosome formation; (iii) movement of negatively charged lipids from the outer leaflet to the inner leaflet of endosomal membrane is electrostatically promoted due to the presence of cationic lipids inside the cavity of endosome, and thus neutral ion pairs are formed, followed by the nucleic acid drugs detaching from cationic liposomes and entering the cytoplasm; (iv) nucleic acid drugs are transfected into cells and perform the corresponding functions within the cell. Cationic liposomes play an important role in the whole process of nucleic acid drugs being delivered into cells, and nucleic acid molecules expressing pathogens (e.g., bacteria and viruses), specific peptides or protein (antigen) fragments of tumor antigens in vivo and finally stimulating specific immune responses. Cationic liposomes are usually composed of cationic lipids, neutral lipids, steroid lipids and PEGylated lipids under appropriate conditions. Cationic lipids and nucleic acids are electrostatically bound together, while co-lipids (neutral lipids and steroid lipids) play a role in preventing lipid oxidation, connecting ligands to the surface of liposomes, or reducing the aggregation of lipid particles. Cationic lipids, generally amphiphilic molecules, have a lipophilic region containing one or more hydrocarbon groups and a hydrophilic region containing at least one positively charged polar headgroup. Cationic lipids and nucleic acids or other giant molecules form a complex bearing overall positive charge, while the surface of cell membrane is overall negatively charged, which makes it easy for nucleic acids and other giant molecules to enter the cytoplasm through the protoplasmic membrane of cells, thus improving the transport efficiency of nucleic acid drugs. Although cationic lipids have made the latest progress in drug delivery, there is still a need for improved cationic lipids that are suitable for regular therapeutic uses in this field. SUMMARY OF THE INVENTION The invention provides novel cationic lipids, cationic liposomes containing the lipids, nucleic acid pharmaceutical compositions containing the liposomes and formulations thereof. The formulations of cationic liposome-nucleic acid pharmaceutical compositions could deliver nucleic acid drugs into cells, improve the transport efficiency of nucleic acid drugs, thereby improving the treatment effect of nucleic acid drugs. The above-desc