Use and constructing plan of anticancer recombined gland virus with tumour CHK1 as target of medicine

Abstract

This invention relates to establishment of artificially reformed human adenovirus 5 and use in treatment of cancers by reorganized configuration of adenovirus. It is obtained by extension of fixed-point deficiency, homologous reorganization, transfection, and monoclone purification of adenovirus. Its novelty includes deletion of 24 alkali groups in encoding sequence of ADV 5 gene set E1A conservative sequence 2, deletion of 28532 - 29360nt of ADV 5 E3 with reversed inserting some of CHK1 cDNA segments. The reorganized configuration of adenovirus can selectively regenerate in tumors, with specific expression of reversed CHK1 cDNA segments to kill tumor cells but normal cells. The configuration is unique effectiveness in diagnosis and treatment of tumors. It provides an ideal therapy of gene target specificity.

Description

1. Technical field [0001] The present invention relates to a human adenovirus type 5 (Human adenovirus type 5, ADV5) recombinant construction scheme, which is characterized in that: directional deletion of the 923-946nt sequence of the ADV5 genome (CTTACC TGC CAC GAG GCT GGC TTT, the sequence codes E1A protein 122 to 129 amino acids), on this basis, further delete the ADV5 E3 region 28532-29360nt, introduce a ClaI restriction site in the deletion region; reversely insert a 620bp exogenous CHK1 cDNA fragment (equivalent to 853-250nt of CHK1 mRNA), so as to obtain the recombinant adenovirus construct Δ923-946ADV5 / ASCHK1 which has a therapeutic effect on tumors. The invention belongs to the technical field of medical genetic engineering. 2. Background technology [0002] Malignant tumors are becoming the primary disease that endangers human health. According to the latest domestic epidemiological survey data, there are more than 3 million cancer patients in my country, about ...

AI Technical Summary

Problems solved by technology

However, the second-generation ADV5 gene therapy vectors have some common disadvantages: (1) Although the vectors designed based on the biological basis of adenoviruses have the therapeutic advantage of replicability in tumor conditions, due to various design deficiencies, the obtained adenoviruses are The efficacy of replication and lysis of tumor cells in tumor cells is far weaker than that of wild-type ADV5. For example, the anticancer effective rate of ONYX-015 alone is only 0-14%

(2) The application of the second-generation ADV5 gene therapy vector is still based on local tumor injection, which is greatly limited in application. In most cases, tumor is a systemic disease and needs to be administered intravenously Systematic treatment is carried out to control primary and metastatic lesions. Therefore, the selectivity of the second-generation ADV5 gene therapy vector and the killing efficiency of tumor cells must be further enhanced to meet the needs of improving drug delivery routes

(3) None of the second-generation ADV5 gene therapy vectors carry effective therapeutic target genes, which limits the therapeutic effect

(4) Since most of the existing molecular targets lack tumor specificity, the strategy of using the second-generation ADV5 gene therapy vector combined with an exogenous promoter, although relatively tumor-selective, still cannot avoid Toxic effects of normal tissue cells

[0010] In addition, many tumor molecular drug targets have been identified in recent years. Designing monoclonal antibodies and small molecular compounds against these molecular drug targets has become the main trend in the development of tumor treatment. Some of them have become prescription drugs, and there are still a large number of molecular therapeutic methods It is in the evaluation stage of clinical trials and has achieved good preliminary results

Nevertheless, most of these new molecular targets and their molecular suppression methods lack tumor specificity, and the resulting dose-limiting toxicity is still a key problem that these therapies generally face and urgently need to be solved, which greatly limits the solved the tumor problem

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