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Novel folate receptor targeted liposome

A folic acid receptor and liposome technology, applied in liposome delivery, genetic material components, gene therapy, etc., can solve the problem of lack of targeting

Inactive Publication Date: 2013-05-01
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Targeted delivery of siRNA in vivo holds important clinical promise, but currently available methods lack targeting and have toxicity concerns

Method used

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  • Novel folate receptor targeted liposome
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Experimental program
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Embodiment Construction

[0024] 1. Materials and Methods

[0025] 1) cell line

[0026] LA-N-5 cell tumors were cultured with 1640 medium containing 10% fetal bovine serum; 2mmol / L L-glutamic acid; 100IU / ml penicillin G; 100mg / mL streptomycin; Box incubation. The medium was changed every 3 days.

[0027] 2) Construction of siRNA

[0028] The siRNA oligonucleotide is aimed at the human MYCN gene, there are two thymidine (DTD) residues at the 3' end, and MYCN-siRNA is labeled with cyanide-3 fluorescence. The sequence is as follows:

[0029] MYCN-1-sense: 5'-CGG AGA UGC UGC UUG AGA Adtdt-3' (SEQ ID NO: 1),

[0030] MYCN-1-antisense: 5'-UUC UCA AGC AGC AUC UCC Gdtdt-3' (SEQ ID NO: 2); (the 26th base of the starting target GI 189234)

[0031] MYCN-2-sense: 5'-GCA GCA GUU GCU AAA GAA Adtdt-3' (SEQ ID NO: 3),

[0032] MYCN-2-antisense: 5′-UUU CUU UAG CAA CUG CUG Cdtdt-3′ (SEQ ID NO: 4); (the 1143th base of the starting target GI 189234)

[0033] MYCN-3-sense: 5'-CAG CAG UUG CUA AAG AAA Adtdt-3' (SEQ ...

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Abstract

The invention discloses a novel folate receptor targeted liposome, which can exclusively inhibit growth of cells of neuroblastoma and consists of distearoylphosphatidylcholine, cholesterol, 1,2-dioleic-3-dimethyl amide propane, polyethylene glycol-CerC16, folate and siRNA of MYCN gene. The liposome has high delivery efficiency in cells of neuroblastoma where a folate receptor is highly expressed and the finally proportion of liquid / oligonucleotides achieves 0.15-0.25 (w / w). The novel liposome maintains the objective siRNA after general administration for a considerably long period of time in blood circulation. The study represents that the targeted liposome carried with siRNA gene, which is served as the product of the invention, is delivered to LA-N-5 cells by the folate receptor, which obviously inhibits MYCN gene expression and cell growth. The liposome has strong capacity of inhibiting cells of neuroblastoma where the folate receptor is over-expressed and can be used as a medicinefor potential targeted treatment of neuroblastoma.

Description

technical field [0001] The present invention relates to novel liposomes and their uses. Based on the high expression of folate receptors in neuroblastoma and the use of RNA interference, the invention discloses a novel liposome targeting folate receptors. And the functional verification of the liposome was carried out by cell biology technology and immunology experiment. Background technique [0002] RNA interference (RNA interference, RNAi) refers to the phenomenon of efficient and specific degradation of homologous mRNA induced by double-stranded RNA (double-stranded RNA, dsRNA), which is highly conserved during evolution. The mechanism is that the double-stranded RNA is degraded by specific nucleases to produce small interfering RNAs (siRNAs). These siRNAs are complementary to homologous target RNAs, and the specific enzymes degrade the target RNAs, thereby inhibiting and down-regulating gene expression. The RNA interference technology developed on this basis refers to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K48/00A61K47/22A61P35/00A61P25/00
Inventor 唐锁勤
Owner GENERAL HOSPITAL OF PLA