CN109700998A - A kind of compound skin injury regeneration renovation agent and preparation method thereof - Google Patents
A kind of compound skin injury regeneration renovation agent and preparation method thereof Download PDFInfo
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Abstract
The present invention is to provide a kind of compound skin injury regeneration renovation agents and preparation method thereof, it is characterized in that, the renovation agent is to be prepared using blood plasma macromolecule protein extract, tea tree root extract and ceramide as major ingredient with borneol, polyethylene glycol (4000-8000), glycerol, ε-poly-D-lysine, carbomer etc. for auxiliary material.The present invention is by a variety of active ingredients such as high molecular reactive protein extract, tea tree root extract and ceramide in animal blood plasma through a kind of compound skin injury regeneration renovation agent made of scientific matching.It, which has, improves skin immunity, improves surface of a wound microcirculation, angiogenesis and granulation tissue is promoted to grow, substantially speed up wound healing speed, realize the effect of surface of a wound perfection is healed without spot.The invention belongs to pure natural biological product, be easy to body absorption, compared with existing products in markets, product of the present invention it is effective in cure it is fast, price is low, easy to use, the advantages that having no toxic side effect, there is good medical Development volue and market application prospect.
Description
Technical field
The invention patent belongs to biomedicine field, and in particular to a kind of compound skin injury regeneration renovation agent and its preparation
Method.
Background technique
Acute and chronic wound is especially the research hotspot that chronic refractory conjunction ulcer is always medical domain.In recent years, although closing
More and more in the research of wound repair, progress is rapid, but under existing treatment level, it is more and more various anxious, slow
Property dermal chronic ulcer allows doctors and patients to be difficult to cope with, and with the development of continuous improvement of people's living standards and regenerative medicine, exploitation can
To promote damage location, structurally and functionally the Regeneration and Repair product of perfect scarless healing and technology have become wound repair
The new research hotspot in field and direction.
Body surface chronic wounds are caused by a series of wounds and disease.It is mainly quiet including traumatic ulcer, lower limb
Arteries and veins ulcer, pressure ulcer, diabetic ulcer etc..Although this kind of surface of a wound rapid causing death unlike cancer, due to
It occurs have the characteristics that the course of disease is long, complication is more, the big body and mind to patient of appearance effects, life and work quality in body surface
It causes great harm.Therefore, the reproducibility reparation for how accelerating skin to damage realizes the surface of a wound from perfection structurally and functionally
No spot repairs oneself problem one of urgently to be resolved as tissue repair field.
The chronic refractory pathogenesis for closing ulcer is caused not yet to illustrate completely so far, doctor trained in Western medicine argument is as follows at present: 1, the surface of a wound
The down regulation of gene expression of extracellular matrix components cytoskeleton causes it to generate reduction, as a result makes the mobile branch of tissue repair cell
Frame goes to pot to delay wound healing.2, in addition to cellular environment, main repair cell is especially fibroblastic to wither rapidly
It dies and further affects extracellular matrix, especially the generation of collagen.3, the influence of local woanded surface growth factor concentration variation, growth
Factor active decline and receptor in target cell structure change with function, combine the factor normally with receptor and obstacle occurs, produce
Raw so-called " losing coupling " phenomenon.4, obstacle occurs for the network of wound healing regulation.Chinese medicine think chronic ulcer of skin be due to
Wound, crush injury, skin trauma etc. have directly injured human body and have experienced malicious evil again, cause QI-blood circulation not normal, lead to skin ulcer,
The protection of skin cannot be carried out again or is detached from environmental stimulation, caused obstinate, seriously endangered patient health.
Clinical treatment doctor trained in Western medicine common antibiotics class at present, not only weak curative effect and also be used for a long time antibiotic can make human body
The microbial balance for relying on, destroying human body is generated, hepatic and renal function can also be damaged.Genomic medicine-was i.e. various in recent years
Porcine HGF enhances skin repair and regenerated function applied to clinically significant, accelerates the effect of wound ulcer healing, but
Also occur many problems in application process.Such as EGF-epidermal growth factor, also known as people oligopeptides -1, it can directly facilitate table
The growth of chrotoplast, with the development of science and technology, detection research discovery EGF is Unidirectional stimulation hyperplasia, even unordered
Ground stimulate hyperplasia, it is this stimulation may result in Skin Cell occur it is difficult to predict the problem of.It is shown according to pertinent literature,
Abuse EGF can also generate side effect, may cause epidermal cell excessively or indeterminate growth, so that cuticula be made to thicken or induce
Other cancerations.Cosmetic surgery hospital and cosmetic industry have occurred much abusing the case of EGF and " disfeaturing " at present.
(bibliography 1, the general outer basis of China and clinical journals the 3rd phase of volume 15 in March, 2008, " epidermal growth factor promoted
The experimental study invaded and shifted into pancreatic cancer cell ", reference paper 2, National Drug Administration on 01 10th, 2019
Publication " EGF cannot function as cosmetic material report ")
Chinese medicine mostly uses Chinese herbal medicine, such as Groomwell root oil, ruyi jinhuang powder, ruyi jinhuang san, powder medicine for removing necrotic tissue and promoting muscle growing to the treatment of chronic ulcer of skin, secondary
Act on small, but indolence, treatment cycle are long and curative effect is not good enough.
Investigation according to the country such as China, Japan, U.S. shows: the hair of chronic wounds patient in comprehensive disease
Sick rate is 7-23%；Such patient is with cardiovascular disease incidence about 41%, with neurological disorders person 27%, in plastic surgery
It is 15%.One investigation display of Britain, the incidence of diabetic ulcer is in gradually increase trend in recent years, is suffered from type II diabetes
In person, ulcer person occurs and accounts for 5.3%；Hospitalizing accounts for 20% to diabetic due to the ulcer of lower limb, wherein amputee about 0.6%.China
The survey showed that for part: ulcer occurrence cause is mainly wound infection (67.5%), pressure ulcer (9.2%), and veins are burst
Ulcer (6.5%), diabetic ulcer (4.9%), other factors (11.9%).
The rather expensive of body surface chronic wounds treatment, it was reported that: the U.S. is used for venous ulcer every year
Medical expense is hundred million dollars of 1.65-10.78,2400 dollars of medical expense of the year of each patient, elderly higher and have year by year
The trend of rise.Therefore, the incidence and social danger of body surface chronic wounds have become modernized society and must face
Major issue.Therefore, seeking the low skin Regeneration and Repair product of a kind of fast curative effect, Small side effects, expense is surgical clinical
It is badly in need of and many patients solves painful serious hope and demand.
Summary of the invention
The purpose of the present invention is based on above-mentioned condition and specializes in a kind of compound skin injury regeneration reparation of exploitation
Agent and preparation method thereof.The present invention is by high molecular reactive protein extract, tea tree root extract and ceramide in animal blood plasma
Etc. a variety of active ingredients through a kind of compound skin injury regeneration renovation agent made of scientific matching.It has raising skin immune
Power improves surface of a wound microcirculation, angiogenesis and granulation tissue is promoted to grow, substantially speeds up wound healing speed, realize wound
The effect of face perfection is healed without spot.
The invention belongs to pure natural biological products, are easy to body absorption, have no toxic side effect, can be widely applied to a variety of causes
Caused wound, burn and scald are burnt and various acute and chronic ulcers etc..Compared with existing products in markets, product of the present invention is effective in cure
Fastly, price is low, easy to use, the advantages that having no toxic side effect.Reparation and regeneration to skin damage, especially to chronic refractory conjunction
The surface of a wound is significant in efficacy, has good medical Development volue and market application prospect.
The purpose of the present invention is achieved through the following technical solutions:
A kind of compound skin injury regeneration renovation agent is with blood plasma macromolecule protein extract, tea tree root extract and neural acyl
Amine is major ingredient, with one of borneol, polyethylene glycol (4000-8000), glycerol, ε-poly-D-lysine, carbomer or a variety of is
Auxiliary material is prepared；Wherein, the accounting of blood plasma macromolecule protein extract is 1.5-10%, and the accounting of tea tree root extract is 10-
30%, the accounting of ceramide is 0.05-0.2%.
The dosage forms such as aqua, paste, freeze-dried powder can be made according to patient and surface of a wound demand in product of the present invention.
The aqua product recipe ingredient mass percent is as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Glycerol 10-20%
ε-poly-D-lysine 0.01-0.5%
Physiological saline surplus, pH7.5-9.0.
The plaster products recipe ingredient mass percent is as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Borneol 1-5%
ε-poly-D-lysine 0.01-0.5%
Glycerol 5-15%
Carbomer 0.1-2%
Physiological saline surplus, pH7.5-9.0.
The freeze dried powder recipe ingredient mass percent is as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Borneol 1-5%
Polyethylene glycol (4000-8000) 5-10%
ε-poly-D-lysine 0.01-0.5%
Physiological saline surplus, pH7.5-9.0.
A kind of preparation method of compound skin injury regeneration renovation agent, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) 20-25% saturation degree is added by volume in fresh animal (pig, ox, sheep etc.) the anticoagulant blood plasma for acquiring sanitary inspection qualification
Ammonium sulfate, stirring are completely dissolved ammonium sulfate, stand 2 hours or so, filter (300-500 mesh) or centrifugation (3000-4000r/
Min, 15-20 points), collection is precipitated as macromolecule protein extract crude product.Filtrate or the heated desalination of centrifugate can do animal egg
White byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: the macromolecule protein extract crude product after inactivation of virus is used in the ratio of 1:4-1:5
Pure water is diluted to solution, and centrifugation (3000-4000r/min, 15-20 points) removal insoluble precipitate is collected centrifugate, will be centrifuged
Liquid carries out hyperfiltration treatment through the ultrafiltration apparatus of molecular cut off 100KD, when retention liquid is concentrated into the 1/4-1/5 of original volume,
The pure water of 4-5 times of trapped fluid volume is added into trapped fluid again, continues the 1/4-1/5 of ultrafiltration desalination to original volume, repeats above-mentioned
Step 2-3 times collects trapped fluid A until sulfate ion is all sloughed in permeate.Recycle the saturating of the liquid containing ammonium sulfate of desalination processes
Cross liquid can be used as next time dissolution macromolecule protein extract crude product lysate reused.
(4) ultrafiltration refines: the ultrafiltration apparatus by the trapped fluid A finally obtained in step (3) again through molecular cut off 300KD
It is concentrated by ultrafiltration, ultrafiltration to trapped fluid volume is the 1/3-1/5 of original volume, and collecting trapped fluid is that blood plasma macromolecule protein mentions
Take object refined liquid (total protein content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.0-7.0 of 8-10 times of gross weight of material adds sodium chloride immersion 24 hours of 10-20% or more.(2) heat is with ultrasonic treatment: anti-
It answers kettle to be heated to keeping the temperature 30-60 minutes in 85~90 DEG C of heat under stiring, room temperature is down to, when carrying out ultrasound cracking (400-500W)
Between 30-60 minutes.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0-12.0, addition 1-5% mixed enzyme (pancreatin:
Cellulase=1:2), the active carbon of 0.5-2% is uniformly mixed, and (revolving speed 130-150r/min) is warming up to 45- with continuous stirring
50 DEG C are hydrolyzed 8-10 hours.
(4) filtering clarification: above-mentioned enzymolysis liquid 100-300 mesh is filtered, filtrate is clarified with sedimentation-type centrifuge again, then is passed through
0.45-0.6 μm of inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product production technology:
1, aqua production technology;
1.5-10% macromolecule protein refined liquid (total protein content >=20mg/ml), 10-30% tea tree are sequentially added by aqua formula
Root extract (general flavone content >=30%, content of tea polysaccharide answer >=10%), 0.05-0.2% ceramide, 0.01-0.5% ε-poly
Lysine is made into active feed liquid by formula rate with the physiological saline of surplus, and active feed liquid is used 1.0 μm respectively, and 0.65 μm,
0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 10-20% through high pressure sterilization glycerol, after mixing, and
With 40% sodium hydroxide tune pH value in 7.5-9.0, aseptic subpackaged, freezen protective.
2, paste finished product production technology:
1.5-10% macromolecule protein refined liquid (total protein content >=20mg/ml), 10-30% tea tree are sequentially added by paste ingredient
Root extract (general flavone content >=30%, content of tea polysaccharide answer >=10%), 0.05-0.2% ceramide, 0.01-0.5% ε-poly
Lysine is made into active feed liquid by formula rate with the physiological saline of surplus, and active feed liquid is used 1.0 μm respectively, and 0.65 μm,
0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, be proportionally added into 10-20% through high pressure sterilization glycerol, 1-5% borneol and
0.1-2% carbomer after mixing, and adjusts pH value in 7.5-9.0, aseptic subpackaged, stored refrigerated.
3, freeze dried powder production technology:
1.5-10% macromolecule protein refined liquid (total protein content >=20mg/ml), 10-30% tea tree are sequentially added by wp formula
Root extract (general flavone content >=30%, content of tea polysaccharide answer >=10%), 0.05-0.2% ceramide, 0.01-0.5% ε-poly
Lysine is made into active feed liquid by formula rate with the physiological saline of surplus, and active feed liquid is used 1.0 μm respectively, and 0.65 μm,
0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into, 1-5% borneol and 5-10% sterilizing polyethylene glycol (4000-
8000) it, after mixing, and adjusts pH value in 7.5-9.0, is put into freeze dryer and is lyophilized, is aseptic subpackaged, it is stored refrigerated.
Product formula mechanism:
Fibrinogen (molecular weight 340KD), fibre are mainly contained in blood plasma macromolecule protein extract in the above formula components
Company albumen (molecular weight 450KD), laminin (molecular weight 804KD), (such as factor VIII molecule amount is greater than coagulation factor
200KD, factor XIII molecular weight 15-30KD) etc. high molecular reactives albumen, although these protein structure, property and
It is different in physiological function, but key effect is all played during the entire process of repair of damaged tissues.They have cell multiplex
Biological activity can stimulate the isocellular generation of epithelial cell, fibroblast, vascular endothelial cell, macrophage, mediate thin
Interaction between born of the same parents and various protein.These macromolecule proteins may participate in the inflammation stress reaction of body complexity, enhancing
To extracellular or cytoplasm protective effect, by forming good coordination with enzymes various in human body in conjunction with receptor corresponding to its
Effect, the permeability for playing hemostasis rapidly, adjusting vascular wall, regulatory protein matter, water, electrolyte diffusion and operating etc..It is same with this
When, they are multi-functional Porcine HGF again, the growth from mesoderm and neuroectodermal cells can be stimulated, such as at fibre
Cell, vascular endothelial cell, epithelial cell, nerve cell, myocyte, osteocyte etc. are tieed up, there is extensive bioactivity.
Pharmacological research shows the three phases in processes of wound repair, i.e. local inflammation reaction stage, cell Proliferation point
Change and granulation tissue formation stages, tissue reconstruction stage, all kinds of blood plasma macromolecule proteins have different degrees of facilitation.?
Inflammatory phase has apparent chemotactic activity to Traumatic cell, can stimulate fibroblast, and vascular endothelial cell etc. is to wound site
It is mobile.When cell migration to damage location, initially enter proliferation and repair the phase, wherein the step of most critical first is that granulation group
The formation knitted.The essence of granulation tissue is a large amount of capillary and fibroblast abundant.Fibrinogen, fine even egg
The macromolecule proteins such as white, laminin, coagulation factor are exactly fibroblast, vascular endothelial cell and vascular smooth muscle cells
Efficient growth accelerator, the formation of new capillary vessel can be strongly facilitated, dramatically increase granulation tissue capillary quantity and
Blood flow improves surface of a wound microcirculation, oxygen necessary to providing for tissue repair and nutriment abundant；Simultaneously to damage group
The regeneration for knitting position nerve fibre also has significant facilitation, can accelerate the recovery of injury tissue function.Numerous studies with face
Bed facts have proved: in wound healing whole process, the bioactivity of these macromolecule proteins can complement each other in body,
It mutually promotes, mutually restrict and form good synergistic effect, make the various cells of body and fiber ordered arrangement and growth, avoid scar
The formation of trace tissue.If any composition purification in them, which is used alone, not only increases production cost, but also can be big
The big effect for reducing wound tissue's healing.So the present invention is by macromolecule protein combined extractings various in blood plasma and synthesis is answered
With, give full play to the team effect of the macromolecule proteins such as fibrinogen, fibronectin, laminin, coagulation factor, realize
The mutual supplement with each other's advantages of various macromolecule proteins participates in comprehensively and mediates three processes of wound repair, promotes the surface of a wound quickly without scar
It repairs.
(bibliography: 2 months 2008 the 2nd phase " fibrin base bio-medicals of volume 22 of Chinese Reconstructive surgery magazine
Material "；
Bibliography: Sichuan physiological science magazine 2002；24(2) " research of human fibrinogen's structure and function "；
Bibliography: " clinical application of plasma fibronectin reparation skin trauma ", Xu Yan Xu Shuan etc., Products in China
Learn magazine the 4th phase of volume 14 in 2001；
Bibliography: foreign medical science physiology, pathology science and clinical fascicle 1998,18 (1) " laminin progress "；
Bibliography: doctor studies the 6th phase of magazine nineteen eighty-three " coagulation factor and mechanism of coagulation "；
Bibliography: " cryoprecipitate clinical application new development " Lei Qianhong, leaf east etc., Nanjing General Hospital, Nanjing Military Area Command, PLA's Blood Transfusion Dept., river
210002 medical graduate students journal of southern Jiangsu capital in January, 2004 the 1st phase of volume 17.)
Formula components tea root is common Chinese herbal medicine simply, and bitter is mild-natured, have heart tonifying diuresis, reducing blood lipid, it is hypoglycemic, antitumor,
Antibacterial anti-inflammatory, antigout and other effects.Clinically it is used for heart disease;Oedema;Hepatitis;Sore swollen toxin;Aphtha;Soup fire burn;It is band-like
Bleb;Psoriasis etc..Mainly containing general flavone, total saposins, procyanidine, tea polysaccharide and polyphenolic substance etc. in tea tree root extract
Ingredient；With antioxidation, the free radicals such as superoxide ion can be removed；Blood viscosity can be obviously improved；Anti-cancer resists and dashes forward
Become；Tea polysaccharide is the important substance that another class has physiological activity in the tea found after tea polyphenols, is a kind of acidoglycoprotein,
And it is combined with a large amount of mineral element.Wherein protein part is mainly made of about 20 kinds of common amino acid, and sugared part is main
By arabinose, xylose, fucose, glucose, galactolipin etc., mineral element is mainly by calcium, magnesium, iron, manganese etc. and on a small quantity micro-
Secondary element, such as rare earth element composition.Pharmacological research shows that tea polysaccharide has hypoglycemic, reducing blood lipid, blood pressure lowering, anticancer, anticoagulant
Blood, antiatherosclerosis, resist oxygen lack, radiation protection, prevention and cure of cardiovascular disease, increases coronary blood flow, enhancing at antithrombotic
The multiple functions such as immunity of organisms.In conclusion in tea tree root extract the various active constituents such as general flavone, tea polysaccharide have it is anti-
Body surface ulcer is repaired in inflammation analgesia, and enhancing immunity of organisms improves the multiple functions such as local blood circulation, can effectively be assisted
The Regeneration and Repair of the chronic wounds of pathogenesis complexity.
(bibliography 1: " anti-inflammatory and analgesic effect of tea root is ground R&D of modern TCM with the 4th phase of volume 26 in 2012 is practiced
Study carefully "；Bibliography 2: Shanghai Chinese magazine the 8th phase in 1992 " corrective action of the tea root to rat plasma lipid "；With reference to
Document 3: 1999 years the 2nd phases " effect of the tea tree root extract to solid tumor " of volume 12 of foreign medical science traditional Chinese medicine fascicle)
Ceramide is the main component for constituting keratoderma lipid, has moisturizing, antiallergy, anti-aging effects, can be with
Whitening is assisted, it is anti-oxidant.Ceramide can form reticular structure in horny layer of epidermis come the hydrone that associates, and keep the water of skin
Point, therefore there is very strong moisture-keeping function to skin；Skin is in aging course, lipid synthesis decline, neural acyl in cuticula
Amine content is reduced, and shows as having dry skin, furfur is coarse, tarnishes；It can be made using ceramide neural in keratoderma
Amide content increases, and can improve the symptoms such as dry skin, furfur, coarse；Meanwhile ceramide can increase skin keratin thickness
Degree improves the moisture holding capacity of skin, reduces wrinkle, increases skin elasticity, it is good that delay skin aging obtains Skin Cell
Restore and regenerates.(bibliography 4: the Chinese medicine o. 11th of volume 36 in November, 2013 " progress of dihydrophytoceramide "；Ginseng
Examine document 5: grain and the grease first phase in 2007 " ceramide acts on skin physiology and its applies in cosmetics ")
Other compositions in formula: borneol, polyethylene glycol (4000-8000), glycerol, ε-poly-D-lysine, Acritamer 940 are
Auxiliary material in product formula.Borneol transmucosal, subcutaneous tissue easily absorb, and there is clearing heat and detoxicating, erosion prevention and muscle growing, swelling and pain relieving to make
With.Polyethylene glycol (4000-8000) and glycerol have lipophilic hydrophily, biocompatibility and nontoxic, non-immunogenicity, and very
Good lubricant, sustained release agent and stabilizer.Water in the also absorbable solution of polyethylene glycol, improves the medical high score with contacting blood
The biocompatibility of sub- material is conventionally used to facilitate cell fusion or protoplast fusion and facilitates organism and takes in conversion
DNA and versatile, can be used for animal and plant and the various cells of microorganism.Carbomer has inherent distinctive cross-linked structure, good
Good viscosity and hydrophilic gel makes it have preferable regulation, slow releasing function to the release of drug.Carbomer has good
Bioadhesion performance, and have hydrogel and nontoxic to body, nonirritant, the hydrogel homogeneous transparent of formation, stability
It is good, it is comfortable on after, it is easy to apply easy to clean.The above auxiliary material is that China's pharmacopeia has been recorded.
ε-poly-D-lysine is a kind of natural antiseptic agent, and antimicrobial spectrum is wide, to gram-positive bacteria, Gram-negative bacteria, ferment
Female bacterium, mould have certain fungistatic effect.ε-poly-D-lysine is not easy the Gram-negative inhibited to other natural antiseptic agents
Escherichia coli, salmonella fungistatic effect it is very good, and its to heat resistance bacillus and it is some virus also have inhibition make
With.
Product of the present invention is made according to the mechanism of action of the above functional component and the performance of various auxiliary materials through scientific matching,
To so far, having no that same document and patent report.
Compared with prior art, the innovative point of the invention patent mainly has:
1, abundant raw material, it can be achieved that resource high value added utilization
Product of the present invention expands the comprehensive of these resources using raw material is wanted based on resourceful animal blood plasma and plant tea root
It closes and utilizes, improve its added value.It is safe and effective and cheap with pure natural, nonirritant, no chemical residues, have
Wide market application prospect, good economic benefit and social benefit.
2, process operability is strong, is convenient for large-scale production
We have been completed from the trial-production to middle and small scale PRODUCTION TRAITS and product stability of laboratory, it was demonstrated that the production technology
Strong operability, at low cost, quality is stable, it can be achieved that large-scale production.What the present invention innovated improves main composition blood
Starch the extraction process that macromolecule protein extract and tea root extract.The technique has no like product and production both at home and abroad at present
Report, so technical advantage is obvious.
Macromolecule protein traditional extraction technique often uses the high production cost such as freeze-thaw method, cryoprecipitate method, affinity chromatography, inconvenient
In large-scale production.This technique combines method with the desalination of different molecular weight cut off loop ultrafiltration and ultrafiltration exquisiteness using saltouing, raw
Produce it is high-efficient, at low cost, convenient for operating, be produced on a large scale.
The pharmaceutical component to work in tea root is mainly general flavone, polysaccharide and its saponin(e metaclass etc., and prior art is a large amount of
Using organic solvents such as alcohol, the pollution to personnel and environment is caused, limits the large-scale production and development and application of the product.
The present invention effectively prevents the highly energy-consuming of conventional organic solvent extraction, heavily contaminated using ultrasound, enzyme, basic hydrolysis process integration
Disadvantage, and it is simple and easy to control, target component yield is high, is produced on a large scale, and is conducive to higher value application tea tree Root Resource.
3, product formula is innovated, and ingredient has complementary advantages, significant in efficacy
Find through our nearly years of researches: blood plasma macromolecule protein extract is combined with tea tree root extract, ceramide answers
With than better effect is used alone.Three kinds of effective ingredient use in conjunction are extracted in conjunction with Chinese traditional herbs extract and animal tissue
The effect of object advantage, complementary shortage can be played, complement each other, mutually promote, the effect that curative effect is stable, selected auxiliary material without
It is malicious, non-stimulated, it is good lubricant, sustained release agent and stabilizer, the bioavilability of functional component and steady can be effectively improved
It is qualitative.We have carried out the follow-up investigation of multiple zoopery and voluntary tested crowd to application effect of products simultaneously, and curative effect is aobvious
It writes, obtains the favorable comment of patient.To so far, which has no that same document and patent are reported.
4, production overall process is environmentally protective
By production procedure of the invention it is found that the leftover bits and pieces generated in each extraction step can comprehensively utilize, other by-products are produced
Product are can be recycled, the discharge without waste water, waste residue substantially, accomplished truly environmentally protective.
Specific embodiment
Following example is only to further illustrate the present invention, and the invention is not limited in any way.
Embodiment 1
Aqua can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma macromolecule egg
White extract 10%, tea tree root extract 30%, ceramide 0.2%, glycerol 20%, ε-poly-D-lysine 0.5%, surplus are made a living
Manage salt water.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) the fresh animal goat-anti blood coagulation slurry for acquiring sanitary inspection qualification, is added 25% saturation degree ammonium sulfate by volume, stirring makes
Ammonium sulfate is completely dissolved, stands 2 hours or so, and 500 mesh filtering, collection is precipitated as macromolecule protein extract crude product.Filtrate warp
Heating desalination can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:5
It is diluted to solution, centrifugation (4000r/min, 20 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/5 of original volume, then 5 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/5 of original volume, repeats the above steps 3 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/5, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH7.0 of 10 times of gross weight of material adds sodium chloride immersion 24 hours of 20% or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
It mixes down and is heated to keeping the temperature 60 minutes in 90 DEG C of heat, be down to room temperature, carry out ultrasonic 60 minutes cracking (500W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH12.0, addition 5% mixed enzyme (pancreatin and cellulose
Enzyme), 2% active carbon is uniformly mixed, and (revolving speed 150r/min) is warming up to 50 DEG C and is hydrolyzed 10 hours with continuous stirring.
(4) filtering clarification: 300 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.6 μ
M inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
10% macromolecule protein refined liquid, 30% tea tree root extract, 0.2% ceramide, 0.5% ε-are sequentially added by aqua formula
Poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 20% through high pressure sterilization glycerol, is uniformly mixed
Afterwards, and adjust pH value 9.0, aseptic subpackaged, freezen protective.
Embodiment 2
Aqua can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma macromolecule egg
White extract 2%, tea tree root extract 10%, ceramide 0.05%, glycerol 10%, ε-poly-D-lysine 0.02%, surplus are made a living
Manage salt water.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) 20 saturation degree ammonium sulfate are added by volume in the fresh animal Niu Kangning blood plasma for acquiring sanitary inspection qualification, and stirring makes sulphur
Sour ammonium is completely dissolved, stands 2 hours or so, is centrifuged (4000r/min, 20 points), and it is thick that collection is precipitated as macromolecule protein extract
Product.The heated desalination of centrifugate can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:4
It is diluted to solution, centrifugation (3000r/min, 15 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/4 of original volume, then 4 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/4 of original volume, repeats the above steps 2 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/3, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.0 of 8 times of gross weight of material adds sodium chloride immersion 24 hours of 10 or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
Under be heated to 85 DEG C heat heat preservation 40 minutes, be down to room temperature, carry out ultrasonic 50 minutes cracking (400W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0, addition 2% mixed enzyme (pancreatin and cellulose
Enzyme), 0.5% active carbon is uniformly mixed, and (revolving speed 130r/min) is warming up to 45-50 DEG C and 8-10 is hydrolyzed with continuous stirring
Hour.
(4) filtering clarification: 100 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.45
μm inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
2% macromolecule protein refined liquid, 10% tea tree root extract, 0.05% ceramide, 0.02% are sequentially added by aqua formula
ε-poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 10% through high pressure sterilization glycerol, is uniformly mixed
Afterwards, and adjust pH value 7.5, aseptic subpackaged, freezen protective.
Embodiment 3
Paste can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma macromolecule egg
White extract 10%, tea tree root extract 30%, ceramide 0.2%, borneol 5%, ε-poly-D-lysine 0.5%, glycerol 15%, card wave
Nurse 2%, surplus are physiological saline.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) 22% saturation degree ammonium sulfate is added by volume in the anticoagulant blood plasma of fresh animal pig for acquiring sanitary inspection qualification, and stirring makes
Ammonium sulfate is completely dissolved, stands 2 hours or so, filters (400 mesh), and collection is precipitated as macromolecule protein extract crude product.Filtrate
Heated desalination can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:5
It is diluted to solution, centrifugation (4000r/min, 20 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/5 of original volume, then 5 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/5 of original volume, repeats the above steps 3 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/5, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.5 of 10 times of gross weight of material adds sodium chloride immersion 24 hours of 15% or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
It mixes down and is heated to keeping the temperature 50 minutes in 90 DEG C of heat, be down to room temperature, carry out ultrasonic 50 minutes cracking (450W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0, addition 4% mixed enzyme (pancreatin and cellulose
Enzyme), 2% active carbon is uniformly mixed, and (revolving speed 150r/min) is warming up to 50 DEG C and is hydrolyzed 10 hours with continuous stirring.
(4) filtering clarification: 200 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.6 μ
M inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
10% macromolecule protein refined liquid, 30% tea tree root extract, 0.2% ceramide, 0.5% ε-are sequentially added by paste ingredient
Poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, be proportionally added into 15% through high pressure sterilization glycerol, 5% borneol and
2% carbomer after mixing, and adjusts pH value 9.0, aseptic subpackaged, stored refrigerated.
Embodiment 4
Paste can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma macromolecule egg
White extract 3%, tea tree root extract 20%, ceramide 0.06%, borneol 1%, ε-poly-D-lysine 0.02%, glycerol 5%, card wave
Nurse 0.5%, surplus are physiological saline.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) 20 saturation degree ammonium sulfate are added by volume in the fresh animal Niu Kangning blood plasma for acquiring sanitary inspection qualification, and stirring makes sulphur
Sour ammonium is completely dissolved, stands 2 hours or so, is centrifuged (3000,15 points), and collection is precipitated as macromolecule protein extract crude product.From
The heated desalination of heart liquid can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:4
It is diluted to solution, centrifugation (3000r/min, 18 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/4 of original volume, then 4 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/4 of original volume, repeats the above steps 2 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/3, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.0 of 8 times of gross weight of material adds sodium chloride immersion 24 hours of 10% or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
Under be heated to 85 DEG C heat heat preservation 30 minutes, be down to room temperature, carry out ultrasonic 30 minutes cracking (400W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0, addition 1% mixed enzyme (pancreatin and cellulose
Enzyme), 0.5% active carbon is uniformly mixed, and (revolving speed 130r/min) is warming up to 45 DEG C and is hydrolyzed 8 hours with continuous stirring.
(4) filtering clarification: 100 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.45
μm inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
3% macromolecule protein refined liquid, 20% tea tree root extract, 0.06% ceramide, 0.02% are sequentially added by paste ingredient
ε-poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, be proportionally added into 5% through high pressure sterilization glycerol, 1% borneol and
0.5% carbomer after mixing, and adjusts pH value 8.5, aseptic subpackaged, stored refrigerated.
Embodiment 5
Freeze dried powder can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma high score
Sub- protein extract 9%, tea tree root extract 25%, ceramide 0.2%, borneol 4%, polyethylene glycol (6000) 10%, ε-poly rely
Propylhomoserin 0.4%, surplus are physiological saline.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) 25% saturation degree ammonium sulfate is added by volume in the fresh animal Niu Kangning blood plasma for acquiring sanitary inspection qualification, and stirring makes
Ammonium sulfate is completely dissolved, stands 2 hours or so, is centrifuged (3500r/min, 20 points), and collection is precipitated as macromolecule protein extract
Crude product.The heated desalination of centrifugate can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:5
It is diluted to solution, centrifugation (4000r/min, 20 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/5 of original volume, then 5 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/5 of original volume, repeats the above steps 3 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/4, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH7.0 of 9 times of gross weight of material adds sodium chloride immersion 24 hours of 15% or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
Under be heated to 88 DEG C heat heat preservation 55 minutes, be down to room temperature, carry out ultrasonic 60 minutes cracking (500W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH12.0, addition 5% mixed enzyme (pancreatin and cellulose
Enzyme), 2% active carbon is uniformly mixed, and (revolving speed 140r/min) is warming up to 50 DEG C and is hydrolyzed 10 hours with continuous stirring.
(4) filtering clarification: 250 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.6 μ
M inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
9% macromolecule protein refined liquid, 25% tea tree root extract, 0.2% ceramide, 0.4% ε-are sequentially added by wp formula
Poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 4% borneol and 10% sterilizing polyethylene glycol
(6000), it after mixing, and adjusts pH value 9.0, is put into freeze dryer and is lyophilized, is aseptic subpackaged, it is stored refrigerated.
Embodiment 6
Freeze dried powder can be made according to patient and surface of a wound demand in product of the present invention.Product specific formula parts by weight are as follows: blood plasma high score
Sub- protein extract 3%, tea tree root extract 12%, ceramide 0.08%, borneol 2%, polyethylene glycol (8000) 6%, ε-poly rely
Propylhomoserin 0.05%, surplus are physiological saline.
Preparation method, comprising the following steps:
(1) blood plasma macromolecule protein extract is prepared
(1) the fresh animal goat-anti blood coagulation slurry for acquiring sanitary inspection qualification, is added 20% saturation degree ammonium sulfate by volume, stirring makes
Ammonium sulfate is completely dissolved, stands 2 hours or so, filters (300 mesh), and collection is precipitated as macromolecule protein extract crude product.Filtrate
Heated desalination can do animal protein byproduct.
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction
It is heated to 60 DEG C in kettle, keeps the temperature 10 hours, inactivation of viruses.
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:4
It is diluted to solution, centrifugation (3000r/min, 15 points) removes insoluble precipitate, centrifugate is collected, by centrifugate through retaining molecule
The ultrafiltration apparatus for measuring 100KD carries out hyperfiltration treatment, when retention liquid is concentrated into the 1/4 of original volume, then 4 is added into trapped fluid
The pure water of times trapped fluid volume continues ultrafiltration desalination to the 1/4 of original volume, repeats the above steps 2 times, until sulfate radical in permeate
Ion is all sloughed, and trapped fluid is collected.The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule egg
The lysate of white extract crude product is reused.
(4) ultrafiltration refines: trapped fluid in step (2) is concentrated by ultrafiltration through the ultrafiltration apparatus of molecular cut off 300KD,
To trapped fluid volume is original volume 1/3, collecting trapped fluid is blood plasma macromolecule protein extract refined liquid (total protein for ultrafiltration
Content >=20mg/ml).
(2) tea tree root extract is prepared
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.0 of 8 times of gross weight of material adds sodium chloride immersion 24 hours of 10% or more.(2) heat is with ultrasonic treatment: reaction kettle is stirring
Under be heated to 85 DEG C heat heat preservation 35 minutes, be down to room temperature, carry out ultrasonic 35 minutes cracking (400W) time.
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0, addition 2% mixed enzyme (pancreatin and cellulose
Enzyme), 0.5% active carbon is uniformly mixed, and (revolving speed 130r/min) is warming up to 45 DEG C and is hydrolyzed 8 hours with continuous stirring.
(4) filtering clarification: 100 mesh of above-mentioned enzymolysis liquid are filtered, filtrate is clarified with sedimentation-type centrifuge again, then through 0.45
μm inoranic membrane refined filtration tea root crude product liquid.Filter residue does feed byproduct for treatment.
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea root purification
Extracting solution (general flavone content >=30%, content of tea polysaccharide answer >=10%).Collecting trapped fluid can be added down in batch enzymolysis step with alkali
Liquid recycles.
(3) finished product is prepared
3% macromolecule protein refined liquid, 12% tea tree root extract, 0.08% ceramide, 0.05% are sequentially added by wp formula
ε-poly-D-lysine is made into active feed liquid by formula rate with the physiological saline of surplus, active feed liquid is used 1.0 μm respectively,
0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 2% borneol and 6% sterilizing polyethylene glycol
(8000), it after mixing, and adjusts pH value 8.5, is put into freeze dryer and is lyophilized, is aseptic subpackaged, it is stored refrigerated.
7 pharmacodynamic experiment of embodiment: compound skin injury regeneration renovation agent of the invention burns using effect to rabbit III degree
The research of fruit.
Experimental animal: 60 common new zealand white rabbits, cleaning grade, rabbit 3~April of age, half male and half female, weight (2.5 ±
0.2) kg purchases Zhengzhou University's Experimental Animal Center, and all rabbits, which mix, divides cage artificial feeding, and environment temperature is 20~22 DEG C, phase
It is 50% to humidity, after artificial mixed fodder is raised 1 week, constructs III degree bum model with ethanol combustion method.
Test grouping and administration: by 60 model rabbits, 5 groups are randomly divided into, experimental group, control group 1, control group 2, control group
3, blank control group, every group 12.It is administered within 24 hours after each group wound, each primary daily morning and evening, each medicament is to be completely covered the surface of a wound
Subject to, the specific situation that is grouped is shown in Table 1:
1 experimental group of table and each control group medicining condition
| Group | Medication | Preparation method |
| Experimental group | Compound skin injury regeneration renovation agent | It is identical with the preparation prepared in embodiment 1 |
| Control group 1 | Fibronectin preparation | Zhengzhou Forberun Biotechnology Co., Ltd. provides |
| Control group 2 | BFGF-ESSEX | It buys from Yisheng Biological Pharmaceutical Co., Ltd., Shuhai |
| Control group 3 | Powder medicine for removing necrotic tissue and promoting muscle growing | Institute of traditional Chinese medicine, Henan Province provides |
| Compare blank group | Physiological saline | 0.85% sterile saline |
Observation index:
Wound healing time: i.e. time required for the complete epithelialization of the surface of a wound.Wound healing criterion is surface of a wound decrustation,
New epithelium is formed completely.Naked eyes are observed continuously 35 days.
Skin healing degree:
The I phase heals: repair place contains a small amount of fibrosed tissue, no scar or a small amount of scar proliferation, and skin corium is clear in structure, complete,
Fibroblast and arrangement of collagen fibers is neat, sebum film is complete.
The II phase heals: for wound repair based on fibr tissue, there are cicatricial contracture or the unordered row of hyperplasia subcutaneous fibrotic in repair place
Column, flesh adipose membrane are imperfect.
The compound skin injury regeneration renovation agent of the invention of table 2 is to rabbit III degree burn treating effect
| Group | Healing time | The I phase heals quantity | The II phase heals quantity |
| Experimental group | 17±1.2 | 9 | 3 |
| Control group 1 | 18±1.5 | 8 | 4 |
| Control group 2 | 22±1.7 | 2 | 10 |
| Control group 3 | 24±0.7 | 3 | 9 |
| Compare blank group | 26±1.3 | 12 | 0 |
From Table 2, it can be seen that compound skin injury regeneration renovation agent of the invention, fibronectin group, bFGF-ESSEX group, dispel it is saprophytic
Flesh dissipates group can shorten wound healing time compared with the control group, have the function of healing after promoting wound, while of the invention
Compound skin injury regeneration renovation agent and fibronectin group also have remarkably promote III degree burn the I phase heal, reduce III degree burning
Hurt the effect of cicatricial tissue hyperplasia.The effect that compound skin injury regeneration renovation agent group of the invention promotes the wound I phase to heal is excellent
In fibronectin group, there were significant differences.Therefore compound skin injury regeneration renovation agent of the invention have promote the surface of a wound without scar again
The raw effect repaired, clinic can be applied to the treatment of all kinds of skin damages.
Claims (9)
1. a kind of compound skin injury regeneration renovation agent, which is characterized in that mentioned with blood plasma macromolecule protein extract, tea root
Taking object and ceramide is major ingredient, in borneol, polyethylene glycol (4000-8000), glycerol, ε-poly-D-lysine, carbomer
It is one or more to be prepared for auxiliary material；Wherein, the accounting of blood plasma macromolecule protein extract is 1.5-10%, and tea root extracts
The accounting of object is 10-30%, and the accounting of ceramide is 0.05-0.2%；The compound skin injury regeneration renovation agent be aqua,
Paste or freeze-dried powder dosage form.
2. a kind of compound skin injury regeneration renovation agent as described in claim 1, which is characterized in that the aqua recipe ingredient
Mass percent are as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Glycerol 10-20%
ε-poly-D-lysine 0.01-0.5%
Physiological saline surplus, pH7.5-9.0.
3. a kind of compound skin injury regeneration renovation agent as described in claim 1, which is characterized in that the paste ingredient component
Mass percent are as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Borneol 1-5%
ε-poly-D-lysine 0.01-0.5%
Glycerol 5-15%
Carbomer 0.1-2%
Physiological saline surplus, pH7.5-9.0.
4. a kind of compound skin injury regeneration renovation agent as described in claim 1, which is characterized in that the freeze dried powder formula
Constituent mass percentage are as follows:
Blood plasma macromolecule protein extract 1.5-10%
Tea tree root extract 10-30%
Ceramide 0.05-0.2%
Borneol 1-5%
Polyethylene glycol (4000-8000) 5-10%
ε-poly-D-lysine 0.01-0.5%
Physiological saline surplus, pH7.5-9.0.
5. a kind of compound skin injury regeneration renovation agent as claimed in claim 1 or 2 or 3 or 4, which is characterized in that the blood
Macromolecule protein extract is starched through the following steps that being prepared:
(1) acquire sanitary inspection qualification fresh animal: pig, ox, sheep anticoagulant blood plasma, by volume addition 20-25% saturation degree sulphur
Sour ammonium, stirring are completely dissolved ammonium sulfate, stand 2 hours or so, and the filtering of 300-500 mesh or centrifugation, collection are precipitated as macromolecule
Protein extract crude product, filtrate or the heated desalination of centrifugate can do animal protein byproduct；
(2) inactivation of virus: by macromolecule protein extract crude product, 3 times of volume pure water is added and are diluted to atherosclerotic, are put into reaction kettle
60 DEG C are heated to, keeps the temperature 10 hours, inactivation of viruses；
(3) loop ultrafiltration desalination: by the macromolecule protein extract crude product after inactivation of virus in the ratio pure water of 1:4-1:5
It is diluted to solution, centrifugation removal insoluble precipitate collects centrifugate, centrifugate is set through the ultrafiltration of molecular cut off 100KD
It is standby to carry out hyperfiltration treatment, when retention liquid is concentrated into the 1/4-1/5 of original volume, then 4-5 times of trapped fluid is added into trapped fluid
The pure water of volume continues ultrafiltration desalination to the 1/4-1/5 of original volume, repeats the above steps 2-3 times, until in permeate sulfate radical from
It is sub all to slough, collect trapped fluid A；The permeate for recycling the liquid containing ammonium sulfate of desalination processes can be used as dissolution next time macromolecule protein
The lysate of extract crude product is reused；
(4) ultrafiltration refines: by the trapped fluid A finally obtained in step (3), the ultrafiltration apparatus through molecular cut off 300KD is carried out again
It is concentrated by ultrafiltration, ultrafiltration to trapped fluid volume is the 1/3-1/5 of original volume, and collecting trapped fluid is blood plasma macromolecule protein extract
Refined liquid, total protein content >=20mg/ml in the extract refined liquid.
6. a kind of compound skin injury regeneration renovation agent as claimed in claim 1 or 2 or 3 or 4, which is characterized in that the tea
Tree root extract is through the following steps that be prepared:
(1) it pulverizes: collecting the tea root raw material of hygienic qualification, crushed, be put into reaction kettle with micronizer, be added former
The water tune pH6.0-7.0 of 8-10 times of gross weight of material adds sodium chloride immersion 24 hours of 10-20% or more；
(2) heat and ultrasonic treatment: reaction kettle is heated to keeping the temperature 30-60 minutes in 85~90 DEG C of heat under stiring, is down to room temperature, into
Row ultrasound pyrolysis time 30-60 minutes；
(3) alkalinity enzymatic hydrolysis: by lysate use 10% sodium hydroxide tune pH11.0-12.0, addition 1-5% mixed enzyme: pancreatin and fibre
Plain enzyme is tieed up, the active carbon of 0.5-2% is uniformly mixed, is warming up to 45-50 DEG C with continuous stirring and is hydrolyzed 8-10 hours；
(4) filtering clarification: above-mentioned enzymolysis liquid 100-300 mesh is filtered, filtrate is clarified with sedimentation-type centrifuge again, then is passed through
0.45-0.6 μm of inoranic membrane refined filtration tea root crude product liquid；Filter residue does feed byproduct for treatment；
(5) ultrafiltration refines: crude product liquid being passed through to the ultrafiltration apparatus of molecular cut off 10,000, permeate is tea tree root extract essence
Liquid processed, extract refined liquid general flavone content >=30%, content of tea polysaccharide answer >=10%；Batch enzymatic hydrolysis can be added down by collecting trapped fluid
It is recycled in step with lye.
7. a kind of preparation method of compound skin injury regeneration renovation agent as claimed in claim 1 or 2, it is characterised in that: should
Renovation agent aqua the preparation method is as follows:
1.5-10% blood plasma macromolecule protein extract, 10-30% tea tree root extract, 0.05- are sequentially added by aqua formula
0.2% ceramide, 0.01-0.5% ε-poly-D-lysine are made into active feed liquid by formula rate with the physiological saline of surplus, will
Active feed liquid uses 1.0 μm respectively, and 0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 10-
20% through high pressure sterilization glycerol, after mixing, and adjusts pH value in 7.5-9.0, aseptic subpackaged, freezen protective.
8. a kind of preparation method of compound skin injury regeneration renovation agent as claimed in claim 1 or 3, it is characterised in that: should
Renovation agent paste the preparation method is as follows:
1.5-10% blood plasma macromolecule protein extract, 10-30% tea tree root extract, 0.05- are sequentially added by paste ingredient
0.2% ceramide, 0.01-0.5% ε-poly-D-lysine are made into active feed liquid by formula rate with the physiological saline of surplus, will
Active feed liquid uses 1.0 μm respectively, and 0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into 10-
20% through high pressure sterilization glycerol, 1-5% borneol and 0.1-2% carbomer, after mixing, and adjusts pH value in 7.5-9.0, and sterile point
Dress, it is stored refrigerated.
9. a kind of preparation method of compound skin injury regeneration renovation agent as described in claim 1 or 4, it is characterised in that: should
Renovation agent freeze dried powder the preparation method is as follows:
1.5-10% blood plasma macromolecule protein extract, 10-30% tea tree root extract, 0.05- are sequentially added by wp formula
0.2% ceramide, 0.01-0.5% ε-poly-D-lysine are made into active feed liquid by formula rate with the physiological saline of surplus, will
Active feed liquid uses 1.0 μm respectively, and 0.65 μm, 0.22 μm of filter membrane carries out refined filtration and miillpore filter degerming, is proportionally added into, 1-5%
Borneol and 5-10% sterilizing polyethylene glycol (4000-8000) after mixing, and adjust pH value in 7.5-9.0, are put into freeze dryer jelly
It is dry, aseptic subpackaged, it is stored refrigerated.
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| CN109700998B (en) | 2022-02-08 |
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