Novel folate receptor targeted liposome

A folic acid receptor and liposome technology, which can be applied in the directions of liposome delivery, genetic material components, and medical preparations of non-active components, etc., can solve the problems of lack of targeting and other problems.

Inactive Publication Date: 2011-05-18
GENERAL HOSPITAL OF PLA
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  • Abstract
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  • Claims
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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
  • Novel folate receptor targeted liposome
  • 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 in 1640 medium containing 10% fetal bovine serum; 2mmol / L L-glutamic acid; 100IU / ml penicillin G; 100mg / mL streptomycin; 37°C, 5% CO2 incubation incubator. The medium was changed every 3 days.

[0027] 2) Construction of siRNA

[0028] The oligonucleotide of siRNA is directed against human MYCN gene, with two thymidine (DTD) residues at the 3' end, and MYCN-siRNA is labeled with cyano-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 1143rd base of the starting target GI 189234)

[0033] MYCN-3-sense: 5'-CAG CAG UUG CUA AAG A...

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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 present invention discloses a novel liposome targeting folate receptors. The liposomes were functionally verified by using cell biology technology and immunological experiments. Background technique [0002] RNA interference (RNAi) refers to a phenomenon that is highly conserved during evolution, induced by double-stranded RNA (dsRNA), and homologous mRNA is efficiently and specifically degraded. The mechanism is that double-stranded RNA is degraded by specific nucleases to generate interfering small RNAs (siRNAs). These siRNAs are complementary to homologous target RNAs, and specific enzymes degrade the target RNAs, thereby inhibiting and down-regulating gene expression. The RNA interference technology developed on this basis refers to the technology in which the double-stranded RNA (...

Claims

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Application Information

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