Preparation and application of analgesic peptide precursor protein ssm-a and its products ssm-a1 and ssm-a2

A technology of less spiny centipede and precursor protein, applied in the field of biomedicine, can solve the problems of insufficient material basis research, achieve good analgesic effect, good clinical application prospect, and simple structure effect

Inactive Publication Date: 2017-03-08
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the material basis research related to its clinical effect of dredging collaterals and relieving pain in centipede is still insufficient

Method used

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  • Preparation and application of analgesic peptide precursor protein ssm-a and its products ssm-a1 and ssm-a2
  • Preparation and application of analgesic peptide precursor protein ssm-a and its products ssm-a1 and ssm-a2
  • Preparation and application of analgesic peptide precursor protein ssm-a and its products ssm-a1 and ssm-a2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Determination of the primary structure of the analgesic peptide precursor protein Ssm-A of the spiny centipede

[0034] 1.1. Transcriptome determination of the venom gland of the less spinous centipede

[0035] Live centipedes were collected from Suizhou, Hubei. Four individuals were randomly selected, and the venom glands of centipedes were isolated at 4 degrees under a stereomicroscope, and immediately put into liquid nitrogen for quick freezing. Then it was transported on dry ice to Shenzhen BGI Institute for sequencing of the venom gland transcriptome. The main experimental steps for the determination of the transcriptome of the commercialized spiny centipede are as follows: extract the total RNA from the venom gland sample of the spiny spiny centipede and use DNaseI to digest the DNA, then use Oligo(dT) magnetic beads to enrich the eukaryotic mRNA; add the interrupting reagent In the Thermomixer, the mRNA is broken into short fragments at an appropriate...

Embodiment 2

[0040] Example 2: Separation, purification and identification of analgesic peptides Ssm-A1 and Ssm-A2 from Centipede spp.

[0041] 2.1. Separation and purification

[0042] Live centipede was collected from Suizhou, Hubei, its venom (dissolved in normal saline) was extracted by electrical stimulation method, vacuum freeze-dried, and stored at -80° for later use.

[0043]The first step: Sephadex G-50 molecular sieve: according to the above method to obtain lyophilized powder of venom, dissolve it in deionized water, take 1ml (protein content is 100mg) and pre-spray it with 20mM Tris-HCl buffer (pH=7.8, containing 0.1M NaCl) to equilibrate the Sephadex G-50 (GE Healthcare, superfine) column (length 122cm, inner diameter width 1.4cm) of 24 hours, carry out elution with the same buffer solution, flow rate is 1.5ml / 10min, collect once every 10min, measure Its absorbance at 280nm, the resulting separation and purification profile is as figure 1 As shown, the analgesic peptides Ssm...

Embodiment 3

[0053] Example 3: Determination of the analgesic activity of the analgesic peptides Ssm-A1 and Ssm-A2

[0054] In order to deeply understand the analgesic activity of the analgesic peptides Ssm-A1 and Ssm-A2 of the spiny centipede, the present invention uses different pain animal models for research, and the results show that the analgesic peptides Ssm-A1 and Ssm-A2 of the spiny centipede are in different All of the animal pain models have shown strong analgesic activity, as follows:

[0055] 3.1. The analgesic effect of the analgesic peptide Ssm-A1 and Ssm-A2 of centipede spp. on the mouse acetic acid writhing pain model:

[0056] As experimental animals, SPF grade Kunming male mice (body weight 18-22 g) were used. The purified analgesic peptides Ssm-A1 and Ssm-A2 were respectively dissolved in sterile saline, and the left side of the experimental animal was intraperitoneally injected (100 μl) with different concentrations of the analgesic peptide Ssm-A1 or Ssm-A2 for 30 min...

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Abstract

The invention relates to preparation of a Scolopendra mutilans analgesic peptide precursor protein Ssm-A and products thereof (Ssm-A1 and Ssm-A2). The Scolopendra mutilans analgesic peptides (Ssm-A1 and Ssm-A2) have the beneficial characteristics of simple structure and special activity. The Scolopendra mutilans analgesic peptides (Ssm-A1 and Ssm-A2) can be separated and purified from Scolopendra mutilans venom. The invention also relates to application of the Scolopendra mutilans analgesic peptides (Ssm-A1 and Ssm-A2); and the Scolopendra mutilans analgesic peptides (Ssm-A1 and Ssm-A2) have analgesic effect on pains caused by multiple factors, and can be used for preparing analgesic drugs.

Description

[0001] Technical field: [0002] The invention relates to the preparation and application of the analgesic peptide precursor protein Ssm-A of centipede spp. and its products Ssm-A1 and Ssm-A2, belonging to the field of biomedicine. Background technique [0003] Centipede is an important traditional animal medicinal material, and its medicinal use has a history of more than 2,000 years. Centipede is warm in nature, poisonous, enters Liver Meridian. Pungent, warm and dry, intense in nature, violent in nature, pervasive in action and expression, can search wind, dredge collaterals and calm convulsions, open phlegm and stagnation, relieve stasis and calm convulsions; it can also quench wind and relieve pain. Detoxification and dissipating stagnation, promoting blood circulation, removing blood stasis and dredging collaterals, tonifying deficiency and promoting impotence, dispelling dampness and dispelling pathogenic factors to detoxify, is a single-use or compound Chinese medicin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435A61K38/17A61P29/00
CPCA61K38/00C07K14/43504
Inventor 张云李文辉杨新旺张悦
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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