Flavivirus replicon constructs for tumour therapy

A flavivirus and replicon technology, applied in the field of expression structure, can solve the problem of unreliable treatment of tumors

Inactive Publication Date: 2008-02-06
レプリカン バイオテク プロプライエタリー リミテッド
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although promising, current systems are not reliable in treating tumors

Method used

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  • Flavivirus replicon constructs for tumour therapy
  • Flavivirus replicon constructs for tumour therapy
  • Flavivirus replicon constructs for tumour therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0165] Construction of KUN replicon expressing murine GMCSF and production of replicon VLPs

[0166] The Kuking replicon Sp6KUNrep4 is obtained by replacing the CMV promoter of the Kuking replicon pKUNrep4 with the SP6 promoter (Varnavski et al., 2000, J Virol 74, 4394-4403), so RNA can be transcribed in vitro relying on SP6 RNA polymerase. Sp6KUNrep4 encodes a puromycin selectable marker, foot-and-mouth disease virus (FMDV) 2A autoprotease (autoprotease), which can cut the inserted foreign protein at the N-terminus, and also contains the internal ribosomal insertion site (IRES) of encephalitis myocarditis virus (EMCV) , which can initiate translation of KUN nonstructural genes essential for RNA replication. The intrinsic ribosomal insertion site (IRES) also takes into account the presence of a stop codon for the foreign gene to ensure that the foreign protein product has a defined C-terminus.

[0167] To further enhance durable RNA replication in cells, cell line-adaptable m...

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Abstract

A flaviviral replicon-based construct is provided for delivery and expression of granulocyte-macrophage colony stimulating factor to facilitate tumour therapy. In particular, the replicon construct encodes a Kunjin virus replicon having one or more mutations in an NS2A non-structural protein that induce enhanced levels of cellular IFN that synergize with recombinant granulocyte-macrophage colony stimulating factor delivered according to the invention. The construct may be administered intra-tumourally or peri-tumourally to an animal as DNA, RNA or packaged into a VLP, for the therapeutic and / or prophylactic treatment of tumours and cancers such as melanoma, lung carcinoma, cervical carcinoma, lung epithelial carcinoma, prostate cancer, breast cancer, renal carcinoma, colon cancer, epithelial cancers and mesothelioma.

Description

technical field [0001] The present invention relates to an expression structure based on a flavivirus replicon, which is used to carry and express granulocyte-macrophage colony-stimulating factor (GMCSF). More particularly, the present invention relates to expression constructs based on Kunjin virus replicons for carrying and expressing GMCSF for tumor treatment. Background technique [0002] GMSCF is a potentially valuable cytokine that can be used in the treatment of cancer. For example, when B16 melanoma cells transfected to express recombinant GMCSF are irradiated with radiation and injected into nude mice, they can be used as a live whole-cell vaccine. These immunized mice were protected against subsequent challenge with B16 tumor cells. These mouse experiments, initially performed to prevent disease, offer the same strategy for testing cancer treatments in humans. [0003] Vaccination with post-irradiated melanoma cells secreting GMCSF enhances the host immune respo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A61K48/00C12N5/10A61K31/7084C12N15/33A61K35/76C12N5/08C12N15/40A61K35/768
CPCC12N2770/24122C12N2770/24123C12N2840/203A61K35/768A61K38/193C07K14/005C12N7/00A61K35/76C12N2770/24132C12N2770/24143A61K35/13C12N15/86C07K14/535A61K38/00A61P35/00A61P35/04A61P43/00
Inventor A·祖尔比尔A·A·赫罗梅赫
Owner レプリカン バイオテク プロプライエタリー リミテッド
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