Gene delivery system and application thereof in preparation of drugs for treatment of tumors
A gene delivery system and applications thereof in the technical field of biological medicines that is particularly useful in the preparation of drugs for treatment of tumors are disclosed. The gene delivery system and applications relate to technology for inducing the differentiation of malignant tumor cells into mature cells, in which regulating the expression of HNF4 alpha protein in the malignant tumor cells using messenger ribonucleic acid, the malignant phenotype of the malignant solid tumor cells is inhibited, and the effective treatment of the malignant solid tumors is achieved. The gene delivery system and applications are therefore applicable to a method of preparing a drug for treating malignant solid tumors and to a method of treating a patient having a malignant solid tumor.
1 . A gene delivery system, comprising a nucleic acid vector and a delivery vehicle, wherein the nucleic acid vector is a self-replicating RNA encoding HNF4 alpha; the gene delivery system delivers the HNF4 alpha or the self-replicating RNA into tumor cells, induces differentiation of the tumor cells into normal mature cells, inhibits proliferation of the tumor cells, and/or induces apoptosis of the tumor cells; the self-replicating RNA has a 35-100 nt polyadenine tail and is an engineered alphavirus genome that includes a 5′-cap, a non-coding or untranslated region (5′ UTR), four genes encoding nonstructural proteins that enable RNA replication, a 26S subgenome promoter, a gene encoding the HNF4 alpha replacing structural protein sequences of the engineered alphavirus genome, and a 3′ non-coding or untranslated region (3′ UTR); and the delivery vehicle comprises lipid-based nanoparticles.
2 . The gene delivery system according to claim 1 , wherein the self-replicating RNA has or carries a 60-75 nt polyadenine tail.
3 . The gene delivery system according to claim 1 , wherein the self-replicating RNA has or carries a 67 nt polyadenine tail.
4 . The gene delivery system according to claim 1 , wherein the self-replicating RNA includes the sequence shown in SEQ ID NO:3.
5 . The gene delivery system according to claim 1 , wherein the lipid-based nanoparticles comprise:
1,2-distearoyl-sn-glycero-3-phosphocholine or 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine, in an amount of ranging from 5% to 20% by moles;
cholesterol, in an amount of ranging from 30% to 55% by moles;
dimyristoylglycerol-polyethylene glycol 2000 or a lipid containing a polyethylene glycol chain, in an amount of ranging from 0.5% to 3% by moles; and
one or more ionizable lipids, in an amount of ranging from 30% to 60% by moles.
6 . The gene delivery system according to claim 5 , wherein the lipid-based nanoparticles have a particle size of 40-300 nm.
7 . A pharmaceutical composition, comprising the gene delivery system according to claim 1 .
8 . The gene delivery system according to claim 5 , wherein the one or more ionizable lipids comprise an N-acyloxyalkyl N-hydroxyalkylamine.
9 . The gene delivery system according to claim 5 , wherein the N-acyloxyalkyl N-hydroxyalkylamine has a formula NR 1 R 2 R 3 , where R 1 is an ω-hydroxy-C 2 -C 10 -alkyl or an ω-hydroxy-C 4 -C 10 -dialkylene ether group, and R 2 and R 3 are independently a C 4 -C 10 -alkylene group substituted with a straight-chain or branched C 10 -C 20 -alkanoyloxy group.
10 . The gene delivery system according to claim 5 , wherein the one or more ionizable lipids comprise N,N-bis [6-(2-hexyldecanoyl)oxyhexyl]-4-hydroxy-butylamine, 8-[(2-hydroxyethyl) [6-oxo-6-(undecyloxy) hexyl]amino]-octanoic acid, 1-octylnonyl ester, or (2-hydroxyethyl)ethylazanediylbis(hexane-6,1-diyl)bis(2-hexyldecanoate).
11 . The gene delivery system according to claim 5 , wherein the 1,2-distearoyl-sn-glycero-3-phosphocholine or 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine is distearoyl-sn-glycero-3-phosphocholine (DSPC), and the dimyristoylglycerol-polyethylene glycol 2000 or the lipid containing a polyethylene glycol chain is dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG 2000).
12 . The gene delivery system according to claim 11 , wherein the DSPC, the cholesterol, the DMG-PEG 2000 and the one or more ionizable lipids are present in a molar ratio of DSPC:cholesterol: DMG-PEG 2000: ionizable lipid(s) of 9.4:42.5:1.8:46.3.
13 . The gene delivery system according to claim 1 , wherein the lipid-based nanoparticles comprise 1,2-distearoyl-sn-glycero-3-phosphocholine or 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine, cholesterol, a lipid containing a polyethylene glycol chain, and one or more ionizable lipids.