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Heaven dragon polypeptide derivative, as well as preparation and application thereof

A Tianlong, natural technology, applied in the directions of biochemical equipment and methods, botanical equipment and methods, applications, etc., can solve the problem that Tianlong anti-tuberculosis pharmacological active ingredients have no relevant literature reports, and achieve the effect of strong killing activity

Inactive Publication Date: 2009-08-05
王辰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Yu Lijun et al.’s patent application “Traditional Chinese Medicine for Treating Tuberculosis and Its Preparation Method” (Patent No. 03132605.6) reported that Tianlong is used to treat scrofula and tuberculosis. Although Tianlong is effective in treating tuberculosis, there is no information about the pharmacological active ingredients of Tianlong against tuberculosis Related literature reports

Method used

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  • Heaven dragon polypeptide derivative, as well as preparation and application thereof
  • Heaven dragon polypeptide derivative, as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: The preparation method of Tianlong antimicrobial peptide

[0027] Crush the dried medicinal materials of Tianlong through an 80-mesh sieve, weigh 2500g of the powder, mix it with 4500ml of distilled water to swell, leave it in a 4-degree refrigerator for 24 hours, centrifuge at low temperature and high speed to obtain 800ml of supernatant, and filter it with a 0.45um microporous membrane to obtain the crude extract. The crude extract was loaded on a solid-phase extraction column ODS-SPE (C18), eluted with water and methanol respectively, and the eluent was lyophilized to obtain a water-soluble part and an alcohol-soluble part, and the alcohol-soluble part was subjected to gel filtration column chromatography The peak where the active ingredient is located is collected, dialyzed and desalted, and separated and purified by reversed-phase high-pressure liquid chromatography to obtain the natural polypeptide.

[0028] in:

[0029] Gel filtration column chromatog...

Embodiment 2

[0034] Embodiment two synthetic antimicrobial peptides are to the mensuration of the minimum inhibitory concentration of Mycobacterium tuberculosis

[0035]1. Medium: 7H9 agar semi-fluid medium: Middlebrook 7H9 dry powder (SIGMA M0178) 4.7g, glycerol 4ml, agar powder 0.5g, add distilled water to 900ml, 15 pounds 121 ℃ autoclave for 10 minutes, add after cooling OADC to 1000ml, set aside.

[0036] Sutong semi-fluid medium: weigh 4.0g sodium glutamate, 1g citric acid, K 2 HPO 4 0.25g, MgSO 4 ·7H 2 0.25g, ferric ammonium citrate 0.025g, glycerin 15ml, agar powder 0.25g, after adding distilled water, heat to make it dissolve completely, after cooling, adjust the pH to 7.2 with 8% NaOH, settle to 500ml with distilled water, pack, Autoclave at 121°C for 20 minutes and set aside.

[0037] 2. Drugs: The control drug is isoniazid; the test drug is a synthetic antimicrobial peptide.

[0038] 3. Strain: Mycobacterium tuberculosis standard strain (Mycobacterium tuberculosisH 37 Rv,...

Embodiment 3

[0044] Example 3 Clinical Observation Curative Effects of Synthetic Antimicrobial Peptides

[0045] Female, 41 years old, was diagnosed with pulmonary tuberculosis in 2000 due to cough and sputum. After treatment with isoniazid, resifopin, pyrazinamide and ethambutol, the condition did not improve significantly, and the drug sensitivity test was polyresistant Drug-refractory tuberculosis (resistant to isoniazid and resifopin), cough, sputum, positive sputum bacteria, and empty lungs. The antibacterial peptide 20 mg was taken orally, bid, and maintained for 3 weeks, the cavity was closed, the sputum bacteria turned negative, and the clinical symptoms disappeared, so the drug was stopped.

[0046] Male, 59 years old. In 2001, due to cough, sputum, fatigue and discomfort, X-ray examination found that he was suffering from cavitary tuberculosis. He was treated with isoniazid, rifampicin, ethambutol and other anti-tuberculosis treatments, but his condition did not improve signific...

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Abstract

The invention relates to a new synthetic antibacterial peptide as well as a preparation method and an application thereof, which belong to the technical field of biomedicine. The antibacterial peptide is a derivative of the natural antibacterial peptide of a dragon and is a single-chain polypeptide further synthesized by a natural polypeptide obtained from the dragon (amphibians in China) by separation. The preparation method comprises the following steps: dry herbs of the dragon are swelled, the sediment is centrifugally removed, the dry herbs are separated and purified after being frozen dry through solid-phase extraction column chromatography, gel filtration column chromatography and reverse-phase high performance liquid chromatography to obtain the natural polypeptide, and the new synthetic antibacterial peptide is designed after the amino acid sequence is determined and the structure is analyzed for the natural polypeptide and is obtained through being expressed by the genetic engineering. The synthetic antibacterial peptide can remarkably inhibit the growth of the mycobacterium tuberculosis including the multi-drug-resistant mycobacterium tuberculosis and can be used for preparing drugs for curing diseases caused by the infection of the mycobacterium tuberculosis including the multi-drug-resistant mycobacterium tuberculosis.

Description

Technical field: [0001] The invention relates to a novel synthetic antibacterial peptide and its preparation method and application, belonging to the technical field of biomedicine. Background technique: [0002] (1) Current status of multi-drug resistant tuberculosis [0003] Tuberculosis is one of the infectious diseases that threaten human health the most. According to WHO statistics, currently about 2 billion people in the world are infected with Mycobacterium tuberculosis, with about 8 million new cases each year and about 3 million deaths, accounting for 7% of the population mortality rate. One of the main reasons for the increase in the incidence and mortality of tuberculosis is the emergence and spread of drug-resistant strains of Mycobacterium tuberculosis. According to reports, about 36% of global tuberculosis patients are infected with drug-resistant bacteria, and this phenomenon is particularly important in developing countries and AIDS patients. In addition, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C12N15/12C12N15/63A61K38/17A61P31/06
Inventor 王辰王鹤尧孔繁翠
Owner 王辰
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