Genetic medicine for treating cancer and other diseases characterized by run-off cell division and its prepn process

A cell division and gene drug technology, applied in gene therapy, drug combination, pharmaceutical formulation, etc., can solve problems such as division and dynamic instability

Inactive Publication Date: 2003-05-14
王晓
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Once assembly or disassembly is affected, dynamic ins

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0015] Extract total RNA from human cells, use the poly(A) RNA as a template to synthesize cDNA with reverse transcriptase, use PCR (polymerase chain reaction) to amplify α-tubulin cDNA, purify, and clone α-tubulin cDNA into The pATub was obtained from the plasmid, and its sequence was verified by sequencing. The oligonucleotide primers of two human α-tubulin genes were synthesized for PCR, wherein the 5'-primer sequence was AATTCGCAAGCTGGC, the 3'-primer sequence was AATGCTGTCAGGTCA, and the 3'-primer spanned the glutamic acid 254 coding region, but The 254th amino acid code is synthesized as 5'TGC3' (its complementary chain is 5'GCA3'), so that it changes from encoding glutamic acid (nucleotide sequence is GAA) to encoding alanine (nucleoside acid sequence is GCA). Use these two primers to carry out PCR synthesis with pATub as a template to obtain a product with a length of 403bp (base pairs), cut it into blunt ends with single-stranded exonuclease, and then digest it with ...

Embodiment 2

[0022] Synthesize oligonucleotide primers of two human α-tubulin genes for PCR, wherein the 5'-primer sequence is AATTCGCAAGCTGGC, the 3'-primer sequence is AATGTCAGGTCAACATT, the 3'-primer spans the glutamic acid 254 coding region, and Make the 254th amino acid code vacant, that is, not synthesize 5'TTC3' (its complementary chain is 5'GAA3' encoding glutamic acid), resulting in the deletion of glutamic acid 254. Use these two primers to carry out PCR synthesis with the pATub obtained in Example 1 as a template to obtain a product with a length of 400bp (base pair), cut it into blunt ends with single-stranded exonuclease, and use a restriction nucleic acid endogenous Dicer ApaLI was digested to obtain a 378bp ApaLI-EcoRI fragment. pATub was digested with restriction endonuclease EcoRI, cut into blunt ends with single-stranded exonuclease, and then digested with restriction endonuclease NcoI to obtain a 165bp EcoRI-NcoI fragment. The ApaLI-EcoRI fragment and the EcoRI-NcoI fra...

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Abstract

The genetic medicine for treating cancer and other diseases characterized by run-off cell division includes carrier and active component, and the active component contains mutant human alpha-tubulin gene with the nucleotide sequence encoding the 254 position amino acid Glu defected or displaced by nucleotide sequence of any other amino acid. The preparation process includes in vitro change of wild human alpha-tubulin gene or its fragment to displace or delete alpha-tubulin Glu 254 and obtain mutant alpha-tubulin gene; and implanting vector in expressable mode. When medicine is guided to the disease part, the expressed mutant alpha-tubulin and wild beta-tubulin are combined to form dimer and assemble microtubule. This will block disassembly of microtubule, break dynamic microtubule instability, disturb mitosis and treat cancer and other diseases characterized by run-off cell division.

Description

Technical field: [0001] The invention relates to a drug for gene therapy for cancer and other diseases characterized by uncontrolled cell division and a preparation method thereof. Background technique: [0002] Disruption or inhibition of cell division is an effective treatment for diseases such as cancer that are characterized by uncontrolled cell division. Spindle is an organelle that undergoes mitosis, while microtubules are the main components of spindles, and microtubules themselves are composed of tubulin. Therefore, if these structures and components are changed or exerted, it is possible to achieve the purpose of interfering with mitosis. Paclitaxel (paclitaxel) has achieved unprecedented success in the treatment of cancer with chemotherapy drugs after it is applied to cancer treatment, which proves that this is a completely feasible way. Paclitaxel binds to β-tubulin, which enhances the stability of microtubules, breaks the normal dynamic instability of microtubu...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61K48/00A61P35/00
Inventor 王晓
Owner 王晓
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