Application of human derived antibacterial peptide LL-37 in preparation of drugs for treating or preventing thrombotic diseases

A human antibacterial peptide, LL-37 technology, applied in the field of biomedicine, can solve problems such as no public reports, and achieve the effects of good antithrombotic activity, inhibition of platelet aggregation, and difficulty in antigen-antibody reaction

Inactive Publication Date: 2015-10-21
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The application of the human-derived antimicrobial peptide LL-37 studied in the present invention in the prevention and / or treatment of thrombotic diseases has been searched in the literature, and there is no public report identical to the present invention.

Method used

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  • Application of human derived antibacterial peptide LL-37 in preparation of drugs for treating or preventing thrombotic diseases
  • Application of human derived antibacterial peptide LL-37 in preparation of drugs for treating or preventing thrombotic diseases
  • Application of human derived antibacterial peptide LL-37 in preparation of drugs for treating or preventing thrombotic diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Determination of the extension of APTT and PT time by human antimicrobial peptide LL-37

[0037] 1) Separate platelet-poor plasma (PPP): Take rabbit carotid blood and place it in a 15ml centrifuge tube containing 1ml of 3.8% sodium citrate (the final volume of each tube is about 10ml, to ensure that the rabbit blood and anticoagulant The volume ratio is 9:1). Centrifuge at 900r / min for 15min. Take the upper layer, which is platelet rich plasma (PRP), and centrifuge the remaining blood sample at 3000r / min for 10min to obtain platelet poor plasma (PPP). .

[0038] 2) APTT time measurement: turn on the coagulometer and warm up to 37°C. And APTT reagent is preheated at 37℃ for 10min, CaCl 2 The solution is also preheated for 15 minutes. First, preheat 20μl PPP and 5μl sample solution at 37°C for 3min in the preheating zone of the coagulometer; then add 25μl APTT reagent to the mixture, and then incubate for 3min at 37°C; change the mixed system into blood After the ...

Embodiment 2

[0047] Example 2 The inhibitory effect of human antimicrobial peptide LL-37 on platelet aggregation

[0048] Healthy human platelet collection (provided by Jiangsu Blood Center) Dilute 1:1 to 2.5×10 with Tyrose`s buffer 8 Pcs / ml, namely platelet rich plasma (PRP), and the remaining healthy human blood platelets were centrifuged at 3200 r / min for 5 min to obtain platelet poor plasma (PPP). For each test, 300μl of PPP is drawn and placed in the test area of ​​the platelet aggregator to zero. During the test, draw 270μl of PRP and place it in the preheating tank, add 20μl of test samples and test beads with different concentration gradients respectively; at 37℃, preheat for 5 minutes and place in the test area; then add 10μl of related inducers such as ADP (Hyphen Biomed, France, final concentration 10μM), thrombin (Hyphen Biomed, France, final concentration 0.26U / ml), U46619 (Sigma-Aldrich, USA, final concentration 0.75μM) or collagen (Sigma-Aldrich, USA, final Concentration 1μg / m...

Embodiment 3

[0051] Example 3 Antithrombotic effect of human antimicrobial peptide LL-37 in rat arteriovenous bypass model

[0052] 1) Experimental animals: 50 SD rats (180-220g), half male and half male, purchased from Qinglongshan Animal Center, Nanjing City, Jiangsu Province.

[0053] 2) Experimental process: The rats were randomly divided into a negative group (saline) and a positive group (aspirin 50 mg / kg), a sample group (5, 10, 15 mg / kg), and a dose of 5 ml / kg. After 15 minutes of tail vein administration, the rats were anesthetized by intraperitoneal injection of chloral hydrate (10% chloral hydrate, 4.5 ml / kg), and an incision was made at the midline of the neck to expose the right carotid artery and left jugular vein. Take two 4cm long PE-50 tubes and one 12cm long PE-60 tube, in which 10cm long cotton thread is built in the PE-60 tube (scraping with a surgical blade to facilitate the formation of thrombus). Then insert a PE-50 tube filled with 50IU / ml heparin into the right carotid...

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Abstract

The invention discloses applications of human derived antibacterial peptide LL-37 in the preparation of drugs for treating and / or preventing thrombotic diseases and functional food for preventing thrombus and coagulation and reducing blood density. The human derived antibacterial peptide LL-37 can inhibit the platelet aggregation, prominently prolongs the APTT and PT time, has an activity for preventing thrombosis, can be used to prepare drugs for preventing and / or treating thrombotic diseases, and can also be used to produce functional food for preventing thrombus and coagulation and reducing blood density.

Description

Technical field [0001] The present invention relates to the field of biomedical technology, in particular to the new medical use of human antimicrobial peptide LL-37 in the preparation of drugs for treating thrombotic diseases and in the preparation of functional foods for preventing thrombosis, anticoagulation and reducing blood viscosity. Background technique [0002] With the continuous improvement of people's living standards, the incidence of high cholesterol, hyperlipidemia, atherosclerosis and the cardiovascular and cerebrovascular thromboembolic diseases caused by them is increasing year by year. Thrombotic diseases are various diseases that cause vascular lumen stenosis and occlusion caused by thrombus, which causes ischemia and infarction of major organs and causes dysfunction. It belongs to cardiovascular and cerebrovascular diseases. The body's normal hemostasis mainly depends on the complete structure and function of the blood vessel wall, effective platelet quality ...

Claims

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Application Information

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IPC IPC(8): A61K38/17A23L1/29A61P7/02A61P9/10
Inventor 孔毅苏雯陈亚会
Owner CHINA PHARM UNIV
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