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Folded chlorotoxin, chlorotoxin mutant, folded chlorotoxin mutant and preparation process thereof

A chlorinated toxin and a preparation process technology, applied in the field of folded chlorinated toxins, can solve the problems of lack of research and innovation of folded structure and variant structure, limited application potential of chlorinated toxins, etc., achieve good economic and social benefits, and realize automatic control , the effect of simple separation and purification process

Inactive Publication Date: 2015-09-02
WENZHOU INST OF BIOMATERIALS & ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current academic research direction is based on chlorotoxins, but there is a lack of research and innovation on the folding structure and variant structure of chlorotoxins. Therefore, the application potential of chlorotoxins is limited.

Method used

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  • Folded chlorotoxin, chlorotoxin mutant, folded chlorotoxin mutant and preparation process thereof
  • Folded chlorotoxin, chlorotoxin mutant, folded chlorotoxin mutant and preparation process thereof
  • Folded chlorotoxin, chlorotoxin mutant, folded chlorotoxin mutant and preparation process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: Chlorotoxin (40 mg) is weighed and placed in a test tube containing 100 mmol of ammonium bicarbonate, 2 mmol of guanidine hydrochloride, 10 mmol of glutathione, and 10% dimethyl sulfoxide mixed solution , vortex at least 3 times until the solution is clear and clear. The system was placed in a refrigerator at 4 degrees Celsius, and the product was taken out at different time points (1 hour, 2 hours, 4 hours, 6 hours, 10 hours, 24 hours). High performance liquid chromatography uses Dionex C18 Acclaim 120 analytical column as the chromatographic column, the flow rate is 1.00 ml / min, and the elution phase is: 0.1% trifluoroacetic acid aqueous solution; 0.1% trifluoroacetic acid acetonitrile solution, 5-65% gradient elution 30 Minutes, get the separation spectrum of the folded chlorinated toxin.

Embodiment 2

[0046] Embodiment 2: Place chlorinated toxin (40 milligrams) in the test tube that 100 millimoles ammonium bicarbonate, 4 millimoles guanidine hydrochloride, 10 millimoles glutathione, 10% dimethyl sulfoxide mixed solution are placed after weighing , vortex at least 3 times until the solution is clear and clear. The system was placed in a refrigerator at 4 degrees Celsius, and the product was taken out at different time points (3 hours, 7 hours, 24 hours). High performance liquid chromatography uses Dionex C18 Acclaim 120 analytical column as the chromatographic column, the flow rate is 1.00 ml / min, and the elution phase is: 0.1% trifluoroacetic acid aqueous solution; 0.1% trifluoroacetic acid acetonitrile solution, 5-65% gradient elution 30 Minutes, get the separation spectrum of the folded chlorinated toxin.

Embodiment 3

[0047] Embodiment 3: after weighing chlorinated toxin (40 milligrams), be placed in the test tube that 100 millimoles ammonium bicarbonate, 2 millimoles guanidine hydrochloride, 100 millimoles glutathione, 10% dimethyl sulfoxide mixed solution are placed , vortex at least 3 times until the solution is clear and clear. The system was placed in a refrigerator at 4 degrees Celsius, and the product was taken out at different time points (3 hours, 7 hours, 24 hours). High performance liquid chromatography uses Dionex C18 Acclaim 120 analytical column as the chromatographic column, the flow rate is 1.00 ml / min, and the elution phase is: 0.1% trifluoroacetic acid aqueous solution; 0.1% trifluoroacetic acid acetonitrile solution, 5-65% gradient elution 30 Minutes, get the separation spectrum of the folded chlorinated toxin.

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PUM

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Abstract

The invention discloses a folded chlorotoxin, a chlorotoxin mutant, a folded chlorotoxin mutant and a preparation process thereof. The peptide sequence of the folded chlorotoxin is MCMPCFTTDHQMARKCDDCCGGKGRGKCYGPQCLCR-NH2. The peptide sequence of folded chlorotoxin mutant is MCMPCFTTDHQMARSCDDCCGGSGRGSCYGPQCLCR-NH2, and the folded chlorotoxin mutant is formed by replacing lysine (Lys, K) inside the peptide sequence of the chlorotoxin with serine (Ser, S). According to the preparation process disclosed by the invention, the potential application value of the chlorotoxin and a ramification thereof in biology and pharmacy can be extended, and good economic benefit and social benefit on the aspects of lift health, individual medical treatment and the like can be achieved.

Description

technical field [0001] The invention relates to a folded chlorinated toxin, a variant of the chlorinated toxin, a variant of the folded chlorinated toxin and a preparation process thereof. Background technique [0002] In recent years, some scholars have discovered a polypeptide consisting of 36 amino acid residues - chlorotoxin (CTX for short). The CTX has good tumor targeting and can specifically bind to various tumor cells, such as glioma, malignant sarcoma, intestinal cancer and prostate cancer. Studies have shown that CTX enters tumor cells through matrix metalloproteinase-2 (MMP-2), and MMP-2 is only highly expressed on the surface of tumor cells, but not on the surface of normal cells, which explains the specific binding of CTX to tumor cells. s reason. At the same time, studies have shown that although CTX is highly toxic to invertebrates, it is not toxic to mammals. At present, 131I-TM-601, a radiotherapy drug modified by CTX, is being approved by the FDA and has...

Claims

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Application Information

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IPC IPC(8): C07K14/00
CPCC07K14/00
Inventor 刘哲
Owner WENZHOU INST OF BIOMATERIALS & ENG
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