CN106361776A - Effect of giant salamander mucus for hemostasis - Google Patents
Effect of giant salamander mucus for hemostasis Download PDFInfo
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- CN106361776A CN106361776A CN201611008091.9A CN201611008091A CN106361776A CN 106361776 A CN106361776 A CN 106361776A CN 201611008091 A CN201611008091 A CN 201611008091A CN 106361776 A CN106361776 A CN 106361776A
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- Prior art keywords
- andrias davidianus
- davidianus blanchard
- mucus
- water
- take
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- 210000003097 mucus Anatomy 0.000 title claims abstract description 82
- 230000023597 hemostasis Effects 0.000 title claims abstract description 19
- 230000000694 effects Effects 0.000 title abstract description 5
- 241000157862 Dicamptodontidae Species 0.000 title abstract 5
- 239000000284 extract Substances 0.000 claims abstract description 30
- 230000000740 bleeding effect Effects 0.000 claims abstract description 29
- 239000003814 drug Substances 0.000 claims abstract description 21
- 206010053615 Thermal burn Diseases 0.000 claims abstract description 9
- 241001253208 Andrias davidianus Species 0.000 claims description 122
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 58
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 58
- 239000013049 sediment Substances 0.000 claims description 32
- 239000008215 water for injection Substances 0.000 claims description 32
- 239000000243 solution Substances 0.000 claims description 26
- 208000032843 Hemorrhage Diseases 0.000 claims description 25
- 238000003756 stirring Methods 0.000 claims description 24
- 239000000706 filtrate Substances 0.000 claims description 23
- 238000001556 precipitation Methods 0.000 claims description 22
- 230000000025 haemostatic effect Effects 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 17
- 238000010438 heat treatment Methods 0.000 claims description 16
- 239000012466 permeate Substances 0.000 claims description 16
- 238000000108 ultra-filtration Methods 0.000 claims description 16
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- 229940030225 antihemorrhagics Drugs 0.000 claims description 14
- 230000001817 pituitary effect Effects 0.000 claims description 12
- 238000004108 freeze drying Methods 0.000 claims description 10
- 108090000145 Bacillolysin Proteins 0.000 claims description 8
- 108010004032 Bromelains Proteins 0.000 claims description 8
- 102000035092 Neutral proteases Human genes 0.000 claims description 8
- 108091005507 Neutral proteases Proteins 0.000 claims description 8
- 239000004365 Protease Substances 0.000 claims description 8
- 206010053476 Traumatic haemorrhage Diseases 0.000 claims description 8
- 208000034158 bleeding Diseases 0.000 claims description 8
- 235000019835 bromelain Nutrition 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- 239000001509 sodium citrate Substances 0.000 claims description 8
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical group O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 6
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000011975 tartaric acid Substances 0.000 claims description 4
- 235000002906 tartaric acid Nutrition 0.000 claims description 4
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 3
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 3
- 239000001630 malic acid Substances 0.000 claims description 3
- 235000011090 malic acid Nutrition 0.000 claims description 3
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 claims description 2
- 235000015165 citric acid Nutrition 0.000 claims description 2
- 230000007062 hydrolysis Effects 0.000 claims description 2
- 238000006460 hydrolysis reaction Methods 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000003292 glue Substances 0.000 claims 1
- 229940079593 drug Drugs 0.000 abstract description 5
- 230000002439 hemostatic effect Effects 0.000 abstract description 4
- 230000000144 pharmacologic effect Effects 0.000 abstract description 2
- 230000006378 damage Effects 0.000 abstract 1
- 238000001356 surgical procedure Methods 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 16
- 230000003301 hydrolyzing effect Effects 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 5
- 239000011664 nicotinic acid Substances 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 241000395633 Andrias japonicus Species 0.000 description 3
- 241000269333 Caudata Species 0.000 description 3
- 241001415306 Cryptobranchidae Species 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 241000269331 Ambystoma Species 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000003032 molecular docking Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 241000269328 Amphibia Species 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 206010053567 Coagulopathies Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001396014 Priacanthus arenatus Species 0.000 description 1
- 208000000260 Warts Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
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- 238000009835 boiling Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- -1 chlorine Aldehyde Chemical class 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 210000004247 hand Anatomy 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
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- 238000007654 immersion Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 230000003533 narcotic effect Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- 230000035943 smell Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/65—Amphibians, e.g. toads, frogs, salamanders or newts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/40—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing ingredients of undetermined constitution or reaction products thereof, e.g. plant or animal extracts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/44—Medicaments
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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- Pharmacology & Pharmacy (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses an effect of giant salamander mucus for hemostasis and application of a giant salamander mucus extract in hemostatic drugs and hemostatic medical devices. The giant salamander mucus extract is used for bleeding caused by external injuries, bleeding caused by burns, bleeding caused by scalds and bleeding caused by surgery; pharmacological experiments indicate that the obtained giant salamander mucus extract has the very good hemostasis effect.
Description
Technical field
The invention belongs to the technical field such as medical science or health food and in particular to a kind of Andrias davidianus Blanchard mucus for stopping blooding
New application.
Background technology
Andrias davidianus Blanchard (andrias davidianus blanchard) is large-scale Amphibia, Caudata, Cryptobranchidae, popular name
" Megalobatrachus japonicus daoidianuas (Blanchard) ", belongs to national second class protection animal, is referred to as " biosphere living fossil ".Cryptobranchidae only deposits 3 kinds of 2 genus at present, wherein
Andrias davidianus Blanchard belongs to 2 kinds: i.e. Chinese giant salamander, Andrias davidianus (andrias davidianus) and Andrias japonicus (andrias japonicus)；The hidden gill
Salamander belongs to cryptobranchid (cryptobranshusalleganiensis) one kind being only distributed in North America.Andrias davidianus Blanchard bisexualism, 7-8
The moon lays eggs on rock, hatching of drifting about in the water.Andrias davidianus Blanchard head width and flat wealthy, the big eye of mouth is little, and extremity are short and small, and skin both sides have bright
Aobvious fold, is distributed wart grain body more.Andrias davidianus Blanchard migrate that ability is poor, big to the dependency of water environment, therefore it is clear in water quality often to dwell
The ground of the dark humidity in the empty more intermountain brook of cool, rock；Its disposition is violent, based on meat, has hibernation more, grows up big
Salamander body length is up to more than 2m.Andrias davidianus Blanchard body surface smooth no squama, aobvious furvous and taupe more than skin, skin has various speckles and rich
Rich gland structure, can secrete white shape mucus in a large number when Andrias davidianus Blanchard is subject to external pessimal stimulation, and originally mucus be in pulpous state, after gradually
It is changed into viscous gum, and with particularity abnormal smells from the patient.Andrias davidianus Blanchard is almost all distributed throughout the country, mainly spreads over the Yangtze river basin
Middle and upper reaches and middle and south mountain area, save distribution with Shaanxi, Hubei, Hunan, Jiangxi, Guizhou etc. and more concentrate.Shan Nan gives birth to as Andrias davidianus Blanchard
, there is the title in " township of Andrias davidianus Blanchard " one of long most preferably suitable living area.In the Chenggu of Hanzhong Area, Nanzheng, Mian County, Yang County, stay
The counties such as dam, Xixiang, Lueyang are widely distributed.At present, Hanzhong Area giant salamander culture has about 2,000,000 tails, and the young salamander of year hatching breaks through 50
More than ten thousand tails, raiser reaches family more than 1500, is that Andrias davidianus Blanchard is laid a good foundation to commercialization, scale, industrialized development.
Andrias davidianus Blanchard whole body is all precious, and the mucus secreted by its epidermis is a particularly important new material, can be used for stopping blooding
Materials application；But developing rapidly with hemostatic material, kind gets more and more, and the constituent of product and structure are also more and more multiple
Miscellaneous.Research shows, the macromolecule hemostatic material toxic and side effects of synthetic are big, and biocompatibility is general, and induces body to produce
Inflammation, pyrogen, anaphylaxiss all more apparent, therefore, carry out in-depth study for Andrias davidianus Blanchard mucus anastalsis, have important
Meaning.
Content of the invention
For these reasons, our researchs obtain a kind of new Andrias davidianus Blanchard pituitary extract, for haemostatic medicament, hemostasis medical
In apparatus, the extraction of the Andrias davidianus Blanchard mucus of the present invention is by Andrias davidianus Blanchard mucus ethanol low-temperature precipitation, and precipitate adjusts ph value, enzymolysis, surpasses
The method of filter, obtains the extract of Andrias davidianus Blanchard mucus；Pharmacological testing shows, the present invention obtains the extract of Andrias davidianus Blanchard mucus, has very
Good anastalsis.
Andrias davidianus Blanchard of the present invention is: raises the Andrias davidianus Blanchard of the second filial obtaining or F3 through artificial bionic state environment.
Andrias davidianus Blanchard mucus of the present invention also known as: Andrias davidianus Blanchard mucus.
The present invention is achieved through the following technical solutions.
A kind of extract of Andrias davidianus Blanchard mucus is for the application in haemostatic medicament, hemostasis medical apparatus；
Wherein haemostatic medicament is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
Wherein hemostasis medical apparatus is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
The preparation method of Andrias davidianus Blanchard mucus includes: Andrias davidianus Blanchard passes through electrostimulation, vacuum method, scrapes rubbing method acquisition Andrias davidianus Blanchard mucus, its
In middle electrostimulation, voltage is 5-10v, vacuum 1kpa-10kpa in vacuum method；The method for preparing extractive of Andrias davidianus Blanchard mucus is:
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of Andrias davidianus Blanchard mucus 1-3 times, heating in water bath, to 30 DEG C -45 DEG C, keeps system temperature
30 DEG C -45 DEG C of degree, stirs 10-30min, adds the ethanol of Andrias davidianus Blanchard mucus 0.5-1.5 times, stirs 5min-10min, from water-bath
Take out, place and stand 8-12 hour under 0 DEG C of -5 DEG C of environment, filter, be precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 30 DEG C -45 DEG C, keeps 30 DEG C -45 DEG C of system temperature, adds
The ethanol of Andrias davidianus Blanchard mucus 3-4 times, stirs 5min-10min, takes out from water-bath, places standing 8-12 under 0 DEG C of -5 DEG C of environment little
When, filter, be precipitated b, precipitation a is merged with precipitation b, is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 2-3 times of Sediment weight, adjusted with ph regulator a
Ph value 4.5-5.5, adds the neutral protease of 0.20-0.40 times of Sediment weight, after hydrolysis 2-4 hour, plus water for injection, use ph
It is 6.0-7.0 that regulator b adjusts ph value, adds the bromelain of 1-2 times of Sediment weight, hydrolyzes 3-6 hour, is digested
Liquid；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 50%-60%, quiet
Put 12-24 hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 4-8 times of Sediment weight, with 0.22 μm of filter element filtering, filter
Liquid with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retains permeate, permeate lyophilization again, obtains Andrias davidianus Blanchard mucus and extracts
Thing.
Wherein ph regulator a is citric acid, malic acid or tartaric acid.
Ph regulator b is sodium citrate.
Andrias davidianus Blanchard of the present invention is: raises the Andrias davidianus Blanchard of the second filial obtaining or F3 through artificial bionic state environment.
Andrias davidianus Blanchard mucus of the present invention also known as: Andrias davidianus Blanchard mucus.
Hemostasis trial is studied
Test 1 external coagulating effectiveness test
Trial drug: present invention preparation embodiment obtains Andrias davidianus Blanchard pituitary extract；
Comparison medicine: take Andrias davidianus Blanchard 1, vacuum method (vacuum 5kpa), obtain Andrias davidianus Blanchard mucus, sterilizing, lyophilization.
Test method: sd rat weighs weight, narcotic amount according to needed for weight calculates, lumbar injection 10% is hydrated chlorine
Aldehyde；After sd rat anesthesia, open thoracic cavity, gather heart blood with syringe, in injection anticoagulant tube, standby；By the basis weighing up
The extract 3mg of inventive embodiments Andrias davidianus Blanchard mucus and positive control drug 10mg adds in test tube, then is added thereto to 2ml anticoagulated whole
Blood solution, is finally added thereto to the cacl of 0.2ml2Solution, starts timing；Observe blood change, until blood occurs substantially
Curdled appearance, stops timing, records clotting time, and 8 Duplicate Samples surveyed by each sample.
Result of the test is shown in Table 1.
The external haemostatic effect comparing result of table 1
Note: compare * p < 0.05 with positive controls.
Conclusion (of pressure testing): above-mentioned test shows, the Blood clotting of the present invention will be significantly better than comparative example, absolutely proves the present invention
The extract of Andrias davidianus Blanchard mucus has more preferable anastalsis
Test 2 mice docking haemostatic effect tests
Trial drug: present invention preparation embodiment obtains Andrias davidianus Blanchard pituitary extract；
Comparison medicine: take Andrias davidianus Blanchard 1, vacuum method (vacuum 5kpa), obtain Andrias davidianus Blanchard mucus, sterilizing, lyophilization.
Test method: weigh the prepared extract sample of Andrias davidianus Blanchard mucus of 0.1g embodiment respectively and 1g positive control drug is big
Salamander mucus lyophilized powder, is respectively placed in displaying bottle, sterilizing is standby；Take the c57bl/6 mice of same batch, weigh, measure tail length；Use handss
After art cuts off 1/3rd of mice tail length, the wound that will dock immediately insertion is shown in bottle, timing simultaneously.When calculating blood flow stopping
Mice amount of bleeding.
Result of the test is shown in Table 2.
Table 2 mice docking haemostatic effect result of the test
Note: compare * p < 0.05 with positive controls.
Conclusion (of pressure testing): above-mentioned test shows, Andrias davidianus Blanchard pituitary extract of the present invention has more outstanding haemostatic effect.
Andrias davidianus Blanchard of the present invention is: raises the Andrias davidianus Blanchard of the second filial obtaining or F3 through artificial bionic state environment.
Andrias davidianus Blanchard mucus of the present invention also known as: Andrias davidianus Blanchard mucus.
A kind of extract of Andrias davidianus Blanchard mucus is for the application in haemostatic medicament, hemostasis medical apparatus；
Wherein haemostatic medicament is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
Wherein hemostasis medical apparatus is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
Preparation embodiment
Embodiment 1
Take Andrias davidianus Blanchard 10, using vacuum method (vacuum 1kpa), obtain Andrias davidianus Blanchard mucus 508g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 1 times of Andrias davidianus Blanchard mucus, heating in water bath, to 30 DEG C, keeps system
30 DEG C of temperature, stirs 10min, adds the ethanol of 0.5 times of Andrias davidianus Blanchard mucus, stirs 5min, takes out from water-bath, places 0 DEG C of environment
Lower standing 8 hours, filters, is precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 30 DEG C, keeps 30 DEG C of system temperature, adds 3 times of Andrias davidianus Blanchard mucus
Ethanol, stir 5min, take out from water-bath, place and stand 8 hours under 0 DEG C of environment, filter, be precipitated b, precipitation a with heavy
Shallow lake b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 2 times of Sediment weight, adjust ph value with citric acid
4.5, add the neutral protease of 0.20 times of Sediment weight, after hydrolyzing 2 hours, plus water for injection, adjust ph value with sodium citrate
For 6.0, add the bromelain of 1 times of Sediment weight, hydrolyze 6 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 50%, stand 24
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 4 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
53.9g (total sugar content is 61.3%).
Embodiment 2
Take Andrias davidianus Blanchard 10, using vacuum method (vacuum 10kpa), obtain Andrias davidianus Blanchard mucus 496g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 3 times of Andrias davidianus Blanchard mucus, heating in water bath, to 45 DEG C, keeps system
Temperature 45 C, stirs 30min, adds the ethanol of 1.5 times of Andrias davidianus Blanchard mucus, stirs 10min, takes out from water-bath, places 5 DEG C of environment
Lower standing 12 hours, filters, is precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 45 DEG C, keeps system temperature 45 C, adds 4 times of Andrias davidianus Blanchard mucus
Ethanol, stir 10min, take out from water-bath, place and stand 12 hours under 5 DEG C of environment, filter, be precipitated b, precipitation a with
Precipitation b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 3 times of Sediment weight, adjust ph value with malic acid
5.5, add the neutral protease of 0.40 times of Sediment weight, after hydrolyzing 4 hours, plus water for injection, adjust ph value with sodium citrate
For 7.0, add the bromelain of 2 times of Sediment weight, hydrolyze 6 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 60%, stand 24
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 8 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
50.6g (total sugar content is 61.3%).
Embodiment 3
Take Andrias davidianus Blanchard 10, using vacuum method (vacuum 5kpa), obtain Andrias davidianus Blanchard mucus 513g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 2 times of Andrias davidianus Blanchard mucus, heating in water bath, to 40 DEG C, keeps system
40 DEG C of temperature, stirs 20min, adds the ethanol of 1 times of Andrias davidianus Blanchard mucus, stirs 8min, takes out from water-bath, places under 3 DEG C of environment
Standing 10 hours, filters, is precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 40 DEG C, keeps 40 DEG C of system temperature, adds Andrias davidianus Blanchard mucus 3.5
Times ethanol, stir 8min, take out from water-bath, place and stand 10 hours under 3 DEG C of environment, filter, be precipitated b, precipitation a with
Precipitation b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 2.5 times of Sediment weight, adjust ph with citric acid
Value 5.0, adds the neutral protease of 0.30 times of Sediment weight, after hydrolyzing 3 hours, plus water for injection, adjust ph with sodium citrate
It is worth for 6.5, adds the bromelain of 1.5 times of Sediment weight, hydrolyze 4.5 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 55%, stand 18
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 6 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
53.7g (total sugar content is 65.0%).
Embodiment 4
Take Andrias davidianus Blanchard 10, using electric shocking method, obtain Andrias davidianus Blanchard mucus 481g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 1.5 times of Andrias davidianus Blanchard mucus, heating in water bath to 35 DEG C, keeping body
It is 35 DEG C of temperature, stirs 15min, add the ethanol of 0.8 times of Andrias davidianus Blanchard mucus, stir 6min, take out from water-bath, place 2 DEG C of rings
Stand 9 hours under border, filter, be precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 35 DEG C, keeps 35 DEG C of system temperature, adds Andrias davidianus Blanchard mucus 3.5
Times ethanol, stir 6min, take out from water-bath, place and stand 9 hours under 2 DEG C of environment, filter, be precipitated b, precipitation a with
Precipitation b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 3 times of Sediment weight, adjust ph value with tartaric acid
4.5, add the neutral protease of 0.25 times of Sediment weight, after hydrolyzing 3 hours, plus water for injection, adjust ph value with sodium citrate
For 6.0, add the bromelain of 1.2 times of Sediment weight, hydrolyze 4 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 55%, stand 16
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 5 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
49.1g (total sugar content is 59.4%).
Embodiment 5
Take Andrias davidianus Blanchard 10, using electric shocking method, obtain Andrias davidianus Blanchard mucus 482g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 3 times of Andrias davidianus Blanchard mucus, heating in water bath, to 45 DEG C, keeps system
Temperature 45 C, stirs 25min, adds the ethanol of 1.4 times of Andrias davidianus Blanchard mucus, stirs 9min, takes out from water-bath, places 4 DEG C of environment
Lower standing 11 hours, filters, is precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 40 DEG C, keeps 40 DEG C of system temperature, adds 4 times of Andrias davidianus Blanchard mucus
Ethanol, stir 9min, take out from water-bath, place and stand 11 hours under 5 DEG C of environment, filter, be precipitated b, precipitation a with heavy
Shallow lake b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 3 times of Sediment weight, adjust ph value with citric acid
5.5, add the neutral protease of 0.35 times of Sediment weight, after hydrolyzing 3.5 hours, plus water for injection, adjust ph with sodium citrate
It is worth for 7.0, adds the bromelain of 2 times of Sediment weight, hydrolyze 5.5 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 60%, stand 20
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 7 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
48.8g (total sugar content is 59.1%).
Embodiment 6
Take Andrias davidianus Blanchard 10, using scraping rubbing method, obtain Andrias davidianus Blanchard mucus 491g；
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of 3 times of Andrias davidianus Blanchard mucus, heating in water bath, to 45 DEG C, keeps system
Temperature 45 C, stirs 25min, adds the ethanol of 1.4 times of Andrias davidianus Blanchard mucus, stirs 9min, takes out from water-bath, places 4 DEG C of environment
Lower standing 11 hours, filters, is precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 40 DEG C, keeps 40 DEG C of system temperature, adds 4 times of Andrias davidianus Blanchard mucus
Ethanol, stir 9min, take out from water-bath, place and stand 11 hours under 5 DEG C of environment, filter, be precipitated b, precipitation a with heavy
Shallow lake b merges, and is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 3 times of Sediment weight, adjust ph value with tartaric acid
5.5, add the neutral protease of 0.35 times of Sediment weight, after hydrolyzing 3.5 hours, plus water for injection, adjust ph with sodium citrate
It is worth for 7.0, adds the bromelain of 2 times of Sediment weight, hydrolyze 5.5 hours, obtain enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 60%, stand 20
Hour, filter, be precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 7 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate
Again with intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract
48.8g (total sugar content is 59.1%).
Total sugar determination method
Sample treatment: take different tests group to obtain Andrias davidianus Blanchard pituitary extract, accurately weighed 0.10g, plus deionized water 10ml,
Dissolving completely, is dissolved in 50ml volumetric flask surely；
The making of standard curve: accurately weigh 0.10g glucose, deionized water constant volume, to 100ml, is configured to standard female
Liquid.The mother solution deionized water constant volume taking different volumes, to 50m l, is made into concentration and is respectively 20 μ g/ml, 40 μ g/ml, 60 μ g/
Ml, 80 μ g/ml, the standard solution of 100 μ g/ml, do blank with deionized water, respectively take 1m l in test tube, add 4ml
Anthrone reagent (2g/l), boils 10min in 100 DEG C of boiling water of immersion, with flowing water cooling after taking-up.On spectrophotometer, enter
Row spectral scan, detection has maximum absorption band in wavelength 620nm, and selects to carry out colorimetric determination at 620nm, and it is bent to draw standard
Line, obtaining formula is y=0.0085x-0.092, r2=0.99.
Sample determination: take 1ml sample in test tube, operated using above-mentioned same step, carry out colorimetric at 620nm
Measure, calculated according to standard curve.
Andrias davidianus Blanchard of the present invention is: raises the Andrias davidianus Blanchard of the second filial obtaining or F3 through artificial bionic state environment.
Andrias davidianus Blanchard mucus of the present invention also known as: Andrias davidianus Blanchard mucus.
A kind of extract of Andrias davidianus Blanchard mucus is for the application in haemostatic medicament, hemostasis medical apparatus；
Wherein haemostatic medicament is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
Wherein hemostasis medical apparatus is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
Andrias davidianus Blanchard of the present invention is: raises the Andrias davidianus Blanchard of the second filial obtaining or F3 through artificial bionic state environment.
Although embodiment of the invention below is described, the invention is not limited in above-mentioned specific embodiment party
Case and application, following specific embodiments are only schematic, guiding, rather than restricted.This area
Those of ordinary skill under the enlightenment of this specification and in the case of the scope protected without departing from the claims in the present invention,
The form of many kinds can also be made, these belong to the row of present invention protection.
Claims (6)
1. a kind of extract of Andrias davidianus Blanchard mucus is for the application in haemostatic medicament, hemostasis medical apparatus.
2. a kind of extract of Andrias davidianus Blanchard mucus according to claim 1 is for answering in haemostatic medicament, hemostasis medical apparatus
With wherein haemostatic medicament is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
3. a kind of extract of Andrias davidianus Blanchard mucus according to claim 1 is for answering in haemostatic medicament, hemostasis medical apparatus
With wherein hemostasis medical apparatus is used for traumatic hemorrhage, burn bleeding, scalds bleeding, surgical operation bleeding.
4. a kind of extract of Andrias davidianus Blanchard mucus according to claim 1 is for answering in haemostatic medicament, hemostasis medical apparatus
With it is characterised in that the preparation method of Andrias davidianus Blanchard mucus includes: Andrias davidianus Blanchard pass through electrostimulation, vacuum method, scrape rubbing method obtain Andrias davidianus Blanchard glue
In liquid, wherein electrostimulation, voltage is 5-10v, vacuum 1kpa-10kpa in vacuum method；The preparation method of Andrias davidianus Blanchard mucus is:
Step (1): take Andrias davidianus Blanchard mucus, add the water for injection of Andrias davidianus Blanchard mucus 1-3 times, heating in water bath to 30 DEG C -45 DEG C, keeping body
It is 30 DEG C -45 DEG C of temperature, stirs 10-30min, add the ethanol of Andrias davidianus Blanchard mucus 0.5-1.5 times, stir 5min-10min, from water
Take out in bath, place and stand 8-12 hour under 0 DEG C of -5 DEG C of environment, filter, be precipitated a, filtrate is standby；
Step (2): take step (1) filtrate, heating in water bath, to 30 DEG C -45 DEG C, keeps 30 DEG C -45 DEG C of system temperature, adds Andrias davidianus Blanchard
The ethanol of mucus 3-4 times, stirs 5min-10min, takes out from water-bath, places and stands 8-12 hour, mistake under 0 DEG C of -5 DEG C of environment
Filter, is precipitated b, and precipitation a is merged with precipitation b, is precipitated；
Step (3): take the precipitation that step (2) obtains, plus the water for injection of 2-3 times of Sediment weight, adjust ph value with ph regulator a
4.5-5.5, adds the neutral protease of 0.20-0.40 times of Sediment weight, after hydrolysis 2-4 hour, plus water for injection, adjusted with ph
It is 6.0-7.0 that agent b adjusts ph value, adds the bromelain of 1-2 times of Sediment weight, hydrolyzes 3-6 hour, obtains enzymolysis solution；
Step (4): take step (3) to obtain enzymolysis solution, add ethanol, make alcohol content in enzymolysis solution reach 50%-60%, standing
12-24 hour, filters, is precipitated；
Step (5): take step (4) to precipitate, add the water for injection of 4-8 times of Sediment weight, with 0.22 μm of filter element filtering, filtrate is again
With intercepting the ultrafiltration membrane ultrafiltration of molecular weight 10000, retain permeate, permeate lyophilization, obtain Andrias davidianus Blanchard pituitary extract.
5. a kind of extract of Andrias davidianus Blanchard mucus according to claim 4 is for answering in haemostatic medicament, hemostasis medical apparatus
With wherein ph regulator a is citric acid, malic acid or tartaric acid.
6. a kind of extract of Andrias davidianus Blanchard mucus according to claim 4 is for answering in haemostatic medicament, hemostasis medical apparatus
With wherein ph regulator b is sodium citrate.
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