Lipid nanoparticle for inhibiting antisense oligonucleotide of bcl-2 and preparation method of lipid nanoparticle

Abstract

The invention discloses a lipid nanoparticle for inhibiting antisense oligonucleotide of bcl-2, and belongs to the field of bio-technology. The lipid nanoparticle for inhibiting the antisense oligonucleotide of bcl-2 is prepared by covering a section of antisense oligonucleotide by virtue of a membrane material, wherein the nucleotide sequence of the lipid nanoparticle is 5'-TCTCCCAGCGTGCGCCAT-3'or 5'-UCUCCCAGCGTGCGCCAU-3'. The invention also discloses a preparation method of the lipid nanoparticle. The nanoparticle provided by the invention can take a good blocking effect on the growth of tumor cells and specific target genes, and especially for KB cervical cancer cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a lipid nanoparticle of antisense oligomeric nucleic acid inhibiting bcl-2 and a preparation method thereof. Background technique [0002] Antisense oligonucleotides are generally composed of 18-22 nucleotides, which selectively bind to target messenger RNA through the principle of complementary base pairing, thereby blocking or inhibiting the function of specific messenger RNA and regulating the subsequent target gene protein expression. [0003] Bcl-2 is a gene that inhibits apoptosis. This gene can promote cell division, expansion, and differentiation, and it is highly expressed in most tumor cells. For the purpose of limiting growth and spreading. G3139 is an antisense oligonucleotide composed of 18 nucleotides. The nucleic acid sequence is 5'-TCT CCC AGCGTG CGC CAT-3'. It can be combined with the messenger RNA encoding bcl-2 through the principle of complementary base pairing....

AI Technical Summary

Problems solved by technology

However, according to previous literature reports, most antisense oligonucleotides lack a suitable drug delivery system, and it is difficult for them to enter cells to play a role by virtue of their own functions. They also have a weak binding effect on target messenger RNA and are difficult to grow. Time keeps inhibiting binding, and most antisense oligonucleotides are easily degraded by nucleases in plasma and lose their therapeutic effect

Although G3139 has a certain curative effect after years of clinical trials, it failed to be approved because the curative effect could not meet the standards of the US FDA.

[0004] Lipid nanoparticle membrane materials in the prior art do not combine Tween (Tween) compounds and polyethylene glycol (PEG) derivatives such as TPGS, if Tween is used alone: ​​Tween has a shorter PEG chain, so Short-chain PEG can increase the repulsion between nanoparticles to prevent their aggregation and reduce stability

On the other hand, due to the lack of long-chain PEG chimera on the surface of nanoparticles, this nanoformulation is easily phagocytized by phagocytes and loses its effect because Tween is easily lost in the systemic circulation.

[0005] Another example is the use of polyethylene glycol (PEG) derivatives such as TPGS alone: ​​Although TPGS has a longer PEG chain, it can avoid the recognition of the phagocytic system to a certain extent, and it will not be engulfed by phagocytes and lose its effect, increasing the long circulation time. However, if there are a lot of TPGS, it will be difficult for the nanoparticles to be effectively taken up by the target tumor cells because of the steric hindrance effect.

[0006] For the antisense oligonucleotide of G3139, most of the previous modifications are phosphorothioated (PS), although this modification can improve the stability of the antisense oligonucleotide to a certain extent, but it is different from the messenger The binding ability, strength and matching degree of RNA are greatly weakened

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