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Lumbricus extract possessing antithrombotic effect

An extract and antithrombotic technology, which is applied in blood diseases, drug combinations, medical raw materials derived from arthropods, etc., can solve the problems of no anticoagulant activity, ineffective extraction of amino acids and nucleic acids, and ease of extraction conditions , to achieve the effect of simple and efficient extraction method

Inactive Publication Date: 2012-03-28
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the homogenized extract can effectively extract protein and better preserve the activity of polypeptide components, the extraction process has not been heated, and the extraction conditions are mild, so that amino acids and nucleic acids cannot be effectively extracted; experiments have shown that ordinary homogenization method The resulting proteinaceous material has thrombolytic activity but not anticoagulant activity

Method used

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  • Lumbricus extract possessing antithrombotic effect
  • Lumbricus extract possessing antithrombotic effect
  • Lumbricus extract possessing antithrombotic effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Analysis of the main chemical components of the extract of Earthworm wet pulverization and the extract of Earthworm decoction.

[0020] 1. Preparation of Dilong wet crush extract: Weigh an appropriate amount of Guangdilong decoction pieces, coarsely pulverize them, and pass through a No. 2 sieve. Accurately weigh 100g of the sieved Earthworm coarsely crushed material, wash away the surface sediment with water, add 1000mL of water, soak for 4 hours, put the material liquid and Earthworm together into a vibrating ultrafine pulverizer (Jinan Beili Powder Co., Ltd. Produced by the company, BL-6 type), crushed for 10 minutes to obtain a slurry with an average fineness D50≤100μm, after 4000r·min -1 Centrifuge, take the supernatant, and pass the resulting supernatant through the membrane pore size 0.45 μm ZrO 2 Ceramic membrane, which can contain crude drug 0.1g·mL -1 The earthworm wet pulverized extract.

[0021] 2. Preparation of the decoction extract of Dilong...

Embodiment 2

[0036] Example 2: Comparison of the antithrombotic efficacy of the ground-wet crushed extract of Earthworm and the decoction of Earthworm.

[0037]The Dilong wet pulverization extract obtained in Example 1 is labeled as sample 1; the obtained Dilong water decoction extract is labeled as Sample 2; , labeled as sample 3; the decoction extract of earthworm water was passed through a polyethersulfone membrane with a molecular weight cut-off of 10,000, and the permeate was taken, labeled as sample 4.

[0038] (1) In vitro anticoagulation experiment:

[0039] Method: On a sterilized ceramic orifice plate, 200 μL of venous blood from healthy SD rats was added to each well, and 200 μL of sample 1, sample 2, sample 3, sample 4, and normal saline (all pre-treated) were added to each well. temperature consistent with the body temperature of the mouse), and finally add 4 μL thrombin (1IU) to each well, mix, put into a constant temperature and humidity incubator at 37°C, and record the co...

Embodiment 3

[0047] Embodiment 3: Comparative test of ordinary homogenate and wet pulverization.

[0048] Operation: After crushing an appropriate amount of earthworm medicinal materials through the No. 2 sieve, take two equal weights of earthworm after sieving, grab water to wash away the surface sediment of earthworm, add 10 times the weight of water, and use Flucker tissue to mix Pulp machine (10000r·min -1 ) and a vibrating ultrafine pulverizer, the homogenized pulverized material and the wet pulverized material were sampled at different time points of 1, 3, 5, 10, 15, and 30 min respectively, centrifuged immediately, and the supernatant was set aside, centrifuged The final dregs are used for particle size measurement.

[0049] (1) Grinding particle size test:

[0050] Re-disperse the centrifuged dregs of the samples obtained at each time point of 1, 3, 5, 10, 15, and 30 minutes after the pulverization time is 1, 3, 5, 10, 15, and 30 min. A parallel determination was performed 3 tim...

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Abstract

The invention relates to a lumbricus extract possessing antithrombotic effect, which takes lumbricus decoction pieces as a raw material. The preparation method comprises the following steps: coarse cracking the raw material of the lumbricus decoction pieces, immersing for 4-5 hours by adding 5-15 times water by weight, adding immersion liquid together in a pulverizer to crush by a wet method so as to obtain slurry with average particle fineness of D50<=100 mum, centrifuging and taking a supernatant, passing through a ceramic membrane with average aperture of 0.45 mum, refrigerating a filtrateat the temperature less than or equal to minus 20 DEG C for 2 hours, and then refrigerating and drying the filtrate under the condition of minus 4 DEG C-minus 6 DEG C of temperature and 0.1mbr-0.05mbr of absolute pressure. The lumbricus extract is capable of extracting amino acid and nucleic acid substances, and keeping the activity of effective chemical ingredients of protein and polypeptide with maximum limit. The lumbricus extract has dual effects of anticoagulation and thromboclasis, animal experiments show that the lumbricus extract has the advantages of good antithrombotic effect and simple extraction method with high efficiency.

Description

technical field [0001] The invention relates to an earthworm extract with antithrombotic effect, which belongs to the field of pharmacy. Background technique [0002] Earth Dragon is the dry product of Pheretima aspergillum (E.Perrier), Pheretima vulgaris Chen, Pheretima.guillelmi (Michaelsen) or Pheretima pectinifera Michaelsen body. The former is commonly called "Guangdilong", and the latter three are commonly called "Shanghaidilong". Dilong, or earthworm, is cold in nature and slightly salty in taste, and is a traditional Chinese medicinal material. According to records, more than 2,000 years ago, our ancestors used earthworm to treat qi deficiency and blood stagnation, meridian stroke caused by blood stasis, hemiplegia and other diseases. There are 40 kinds of earthworm prescriptions listed in "Compendium of Materia Medica". Among these prescriptions, earthworms are extracted by decoction or powdered and directly taken orally, and the curative effect is remarkable. ...

Claims

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Application Information

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IPC IPC(8): A61K35/64A61P7/02A61K35/62
Inventor 郭立玮付廷明杨丰云
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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