CN101548935A - Use of acidic fibroblast growth factor in preparing cosmetics - Google Patents
Use of acidic fibroblast growth factor in preparing cosmetics Download PDFInfo
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- CN101548935A CN101548935A CNA2009100669522A CN200910066952A CN101548935A CN 101548935 A CN101548935 A CN 101548935A CN A2009100669522 A CNA2009100669522 A CN A2009100669522A CN 200910066952 A CN200910066952 A CN 200910066952A CN 101548935 A CN101548935 A CN 101548935A
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Abstract
The present invention discloses a user of acidic fibroblast growth factor in preparing cosmetics, and simultaneously provides a production process. The invention uses biotechnology for beautifying. The acidic fibroblast growth factor is used for external beautifying. The physiologic status of skin is improved at root and the restoring and metabolism of epidermic tissue are promoted through complementing the biological active factors which promote the metabolism and proliferation of epidermic cell. The skin can be smoothed and young. The color spot is faded and the wrinkle is pacified. The damage of sunshine to the skin is reduced. The skin smoothing and beautifying effect is obvious. Furthermore the acidic fibroblast growth factor is safe and reliable without side effect.
Description
Technical field
The invention provides the new purposes of acid fibroblast growth factor, especially disclose the purposes of acid fibroblast growth factor in the preparation cosmetics, its production technology also is provided simultaneously, belong to the biological beauty product technique field.
Background technology
The acid fibroblast growth factor that the present invention relates to (Acid Fibroblast Growth Factor is called for short aFGF) is by the secreted a kind of short cell division somatomedin of body various kinds of cell.AFGF can stimulate and regulate multiple mesoderm, the neuroectodermal cell differentiation propagation of originating from such as vascular endothelial cell, epithelial cell, sarcoplast, osteoblast and neurogliocyte, plays an important role in fetal development, organization healing.
The research of aFGF at present or application all are confined to promote the reparation aspect of damage (especially nerve injury), and Shang Weijian has its research or application aspect beauty treatment.
Summary of the invention
The invention discloses the purposes of a kind of acid fibroblast growth factor in the preparation cosmetics.
The present invention also provides the cosmetics of being made by acid fibroblast growth factor, has the effect of beauty and skin care.
Technical solution of the present invention comprises:
1, with the acid fibroblast growth factor is main active, adopts special vacuum freeze-drying technique and conventional cosmetics production technology to make cosmetics.
2, a kind of aFGF of containing also is provided is the cosmetics (ratio of weight and number) of main active in the present invention:
AFGF 1～100,000 AU
Active stabilizer: 4～8g
Accelerative activator: 0.3～3g
Protease inhibitor: 0.1～0.6g
Percutaneous absorption enhancer: 0.1～2g
Used protease inhibitor is any one in boric acid leucine, boric acid valine, boric acid alanine, the N-acetylcystein.
Used Percutaneous absorption enhancer is any one in laurocapram, propylene glycol, the calcium mercaptoacetate.
Used active stabilizer is any one of hyaluronic acid, mannitol, dextran;
Used activity promotes that into agent is a water-solubility collagen.
The preparation method of described cosmetics may further comprise the steps:
In aFGF, add active stabilizer, accelerative activator, protease inhibitor and Percutaneous absorption enhancer any one or multiple, be mixed with aqueous solution with purified water, adopt vacuum freeze-drying technique to make lyophilized powder again.
Relate to following composition in the technical scheme of the present invention:
Active component: aFGF
Active stabilizer: hyaluronic acid, mannitol, dextran;
Accelerative activator: water-solubility collagen;
Protease inhibitor: boric acid leucine, boric acid valine, boric acid alanine, N-acetylcystein;
Percutaneous absorption enhancer: Zuo En (laurocapram, Azone), propylene glycol, calcium mercaptoacetate.
In above-mentioned active component, add in active stabilizer, accelerative activator, protease inhibitor and the Percutaneous absorption enhancer any one or multiple, be mixed with aqueous solution with cryogenic purincation water, and place ice bath, to be packed.With above-mentioned preparation and aqueous solution, divide to install in the cillin bottle, place vacuum freeze drier to carry out after the lyophilizing promptly.
The manufacturing of special solvent
0.1～2 part of Percutaneous absorption enhancer
0.1～0.6 part of protease inhibitor
4～8 parts of active stabilizers
2～6 parts of glycerol
PH5.0～7.4PBS solution adds to 100 parts
According to the above ratio Percutaneous absorption enhancer, protease inhibitor, active stabilizer are mixed,, add glycerol with the dissolving of PBS solution, mixing, packing is jumped a queue, with behind the flowing steam sterilization promptly.
The preparation manufacturing of lyophilized powder
With active component, active stabilizer, accelerative activator and/or active stabilizer and/or protease inhibitor and/or Percutaneous absorption enhancer, be mixed with aqueous solution according to the above ratio, and place ice bath with cryogenic purincation water; With above-mentioned preparation and aqueous solution, divide to install in the cillin bottle, with plug false add plug, place vacuum freeze drier to carry out lyophilizing.
Freeze-dry process and condition are: open vacuum freeze drier, goods are carried out pre-cooling, products temperature is reduced to-60 ℃, temperature fall time is 4-5 hour.Start vacuum pump, vacuum in the case is evacuated to 0.3Mbar.Heat, distillation, the distillation time is 14-18 hour, and distillation finishes, and tamponade charges into aseptic gas in the case, unpack, and takes out goods, rolls lid, warehouse-in.
Product form is that lyophilized powder adds special solvent, needs during use to make lytic agent with special solvent.
The prescription of special solvent and technology are:
With Percutaneous absorption enhancer, protease inhibitor, active stabilizer, with pH5.0～7.4PBS solution dissolving, add glycerol, mixing, packing is jumped a queue, with 100 ℃ of flowing steam sterilizations after 30 minutes promptly.
Active stabilizer can be used wherein one or more, but three kinds of active stabilizers and a kind of accelerative activator while result of use the best.Percutaneous absorption enhancer and protease inhibitor be each general only a kind of with wherein in prescription, also can severally share, and its effect is similar.
Below experiment shows cosmetic result of the present invention:
1 material
Active component: aFGF is commercially available product, also can adopt gene engineering research to produce voluntarily.Will carry out determination of activity before using, what the aFGF determination of activity was adopted is cell proliferation/mtt assay.
Active stabilizer: hyaluronic acid, mannitol, dextran.Commercially available, chemical pure.
Accelerative activator: water-solubility collagen, commercially available, purity is greater than 95%.
Protease inhibitor: boric acid leucine, boric acid valine, boric acid alanine, N-acetylcystein, commercially available, chemical pure.
Percutaneous absorption enhancer: Zuo En (laurocapram, Azone), propylene glycol, calcium mercaptoacetate.Commercially available, chemical pure.
Protease: pepsin, trypsin, commercially available, chemical pure.
2 methods
2.1 the manufacturing of freeze-dried powder preparation: referring to above
2.2 the manufacturing of special solvent: referring to above
2.3 protease inhibitor degraded test
Experiment product: the freeze-dried powder preparation that contains active component of above-mentioned manufacturing is dissolved with special solvent, makes aqueous solution, and measure the activity of its active component, with this activity value as 100%.
Reference substance: except that not containing the protease inhibitor all the other same experiment products.Make aqueous solution with purified water, and measure the activity of its active component, with this activity value as 100%.
Protein enzyme solution:, be made into the tryptic mixed aqueous solution that contains 1% pepsin and 1% with purified water with pepsin and trypsin.
Get each 20ml of experiment product and reference substance respectively, add protein enzyme solution 0.5ml respectively, mixing is put in 37 ℃ of water-baths and is hatched, and respectively at different time samplings, carries out determination of activity.
2.4 stability test
Get the above-mentioned freeze-dried powder preparation that contains active component that makes, place room temperature and 7 ℃ of refrigerators respectively, after placing the different time, take a sample, carry out content and determination of activity.
2.5 cosmetic result test
Experiment product: the lyophilized formulations that contains aFGF
Blank product: the lyophilized formulations that does not contain aFGF
Commercially available reference substance: commercially available certain brand skin care wrinkle removing cosmetics
Experiment product and blank product all with the special solvent dissolving, afterwards, are coated skin of face before use, and are carried out the skin importing with specific method (having declared utility model patent).Carry out following cosmetic result test respectively:
2.5.1 skin care, wrinkle removing test
Choose 40～55 years old old people 120 people, the men and women half and half.Be divided into 3 groups at random, every group 40 people, the men and women half and half.Every group is used above-mentioned experiment product or reference substance respectively, continuous 2 months, estimates the cosmetic result of its skin care, wrinkle removing then respectively.
2.5.2 speckle dispelling test
Choose 20～35 years old facial women 120 people that mottle is arranged, be divided into 3 groups at random, every group 40 people.Every group is used above-mentioned experiment product or reference substance respectively, continuous 2 months, estimates its freckle effect then respectively.
2.5.3 anti-acne test
Choose 18～25 years old facial people 80 people that comedo is arranged, the men and women half and half, is divided into 2 groups at random, every group 40 people, and the men and women half and half.Every group is used above-mentioned each experiment product or blank product respectively, continuous 2 months, estimates its anti-acne effect then respectively.
2.5.4 safety testing
In above-mentioned skin care wrinkle removing test, speckle dispelling test and anti-acne test, observe its zest, anaphylaxis and other untoward reaction simultaneously to skin.
3 results
3.1 the manufacturing result of freeze-dried powder preparation
Through the produced freeze-dried powder preparation of above-mentioned technology, outward appearance is loose bulk or powdery, is white in color or off-white color.Add water electrode and easily dissolve, be the solution of clear after the dissolving.Detect through microbiology, do not detect microorganism alive, be sterile solution.The sanitary standard that surpasses the relevant cosmetics of country.
3.2 the manufacturing result of special solvent
Through the produced special solvent of above-mentioned technology, outward appearance clear.Detect through microbiology, do not detect microorganism alive, be sterile solution.The sanitary standard that surpasses the relevant cosmetics of country.
3.3 protease inhibitor degraded result of the test
Table 1 protease inhibitor degraded result of the test
As can be seen from the above table, experimental group under 37 ℃ of temperature with pepsin and trypsin combined effect after 30 minutes, its activity does not descend substantially, acts on after 1 hour, its activity still reduces seldom.And matched group with protease combined effect after 10 minutes, it is active promptly only surplus original below 1/5th.Presentation of results, the present invention's goods have good protease inhibitor Degradation.And when carrying out beauty and skin care with this product, goods on skin stop and the time of importing be no more than 10 minutes, so the time of the protease inhibitor Degradation of the present invention's goods is enough.
3.4 stability test result
Table 2 stability test result
As can be seen from the above table, the present invention's goods, room temperature place and 7 ℃ under when being placed to 36 months, its content there is no significant change; When being placed to 18 months at normal temperatures, its activity is not seen significant change; When being placed to 24 months under 7 ℃, its activity is not seen significant change yet.The result shows, the present invention's goods can be preserved 18 months at normal temperatures, as placed below 7 ℃, then the holding time longer, be more than 2 years.
3.5 cosmetic result result of the test
3.5.1 skin care, wrinkle removing result of the test
Table 3 skin care, wrinkle removing result of the test
As can be seen from the above table, aFGF has good skin care wrinkle removing effect; The blank group that does not contain aFGF does not have the effect of skin care wrinkle removing; The cosmetic result of aFGF group all significantly is better than commercially available reference substance group.
3.5.2 speckle dispelling result of the test
Table 4 speckle dispelling result of the test
As can be seen from the above table, the aFGF group has freckle effect preferably.
3.5.3 anti-acne result of the test
Table 5 anti-acne result of the test
As can be seen from the above table, the aFGF group has anti-acne effect preferably.
3.6 safety testing result
Table 6 safety testing result
After 320 people used, not finding had zest to skin, only found that 7 human faces are slightly irritated, had 2 people parts to itch or flushing.
When the present invention is applied to aFGF to improve looks, solved three key difficult problems, otherwise it can't practical application.That is:
(1) aFGF reaches at normal temperatures and is difficult to keep its biologic activity in the ordinary cosmetics dosage form.After placing the short time at normal temperatures, it promptly loses its biologic activity, so its production, transportation, preservation all become insurmountable barrier, makes its practical application be difficult to realize.In addition, its performance biologic activity needs specific pH value, ionic strength and temperature, so make common cosmetics dosage form such as cream, it will not have any biologic activity.
(2) aFGF is easily lost activity by proteasome degradation.The chemical nature of aFGF is a protein and peptide, and there is a large amount of protease in the application on human skin surface, behind aFGF contact skin, can be lost activity by the proteasome degradation of skin surface very soon.
(3) aFGF is difficult to Transdermal absorption.AFGF all is macromole bioactive substances, and under normal circumstances, it is difficult to see through skin and absorbs.
The present invention has successfully solved an above-mentioned difficult problem, makes aFGF be applied to beauty treatment and becomes a reality.
Beneficial effect of the present invention is: aFGF is used for the external beauty treatment, by replenishing the bioactie agent that promotes cellular metabolism and propagation, fundamentally improves the physiological status of skin, promote the reparation and the metabolism of skin histology; Make skin abnormality delicate young; The desalination mottle calms down wrinkle; Alleviate the damage of Exposure to Sunlight to skin.So the skin-softening beauty effect is remarkable, and safe and reliable, has no side effect.
Percutaneous absorption enhancer is applied in the beauty treatment prescription of aFGF, has solved the problem that aFGF is difficult to Transdermal absorption, improved its cosmetic result significantly.
Protease inhibitor is applied in the beauty treatment prescription of aFGF, has solved aFGF, kept the activity of aFGF, promoted the absorption of aFGF, improved the effect of its skin-softening beauty significantly easily by the problem of proteasome degradation.
AFGF is made freeze-dried powder preparation, solved its easy at normal temperatures deactivated problem.The freeze dried powder that this prescription is made can keep active 18 months at normal temperatures; In 0～7 ℃ of freezer, can keep active more than 2 years.But so room temperature transportation and preservation, as place freezer or refrigerator to keep, effect is better.Can't keep active problem when having solved aFGF production, transportation and preservation, the practical application of aFGF is become a reality.
Its manufacturing process is reasonable, is suitable for large-scale industrialization production.
The specific embodiment
By following examples the present invention is described for example further, and do not limit the present invention in any way, under the prerequisite that does not deviate from technical solution of the present invention, any change or change that those of ordinary skills that the present invention did are realized easily all will fall within the claim scope of the present invention.
Main material:
Active component: aFGF.Can make voluntarily, also buyable is purchased acquisition.Its main processes of making voluntarily is: the genetic engineering bacterium of aFGF is fermented in fermentation tank, and the centrifugal thalline of abandoning is got supernatant, saltouts.Centrifugal, get precipitation.To precipitate redissolution, carry out purification, collect the aFGF peak, put in-20 ℃ of low temperature refrigerators frozen with chromatography.Purity and determination of activity are carried out in sampling.
Active stabilizer: hyaluronic acid, mannitol, dextran, commercially available, chemical pure.
Accelerative activator: water-solubility collagen, commercially available, purity is greater than 95%.
Protease inhibitor: boric acid leucine, boric acid valine, boric acid alanine, N-acetylcystein, commercially available, chemical pure.
Percutaneous absorption enhancer: Zuo En (laurocapram, Azone), propylene glycol, calcium mercaptoacetate.Commercially available, chemical pure.
Embodiment 1:
Measure aFGF 1～20,000 AU, hyaluronic acid 1～6g, mannitol 3～8g, dextran 3～8g, water-solubility collagen 1～6g, purified water adds to 100ml; Above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Embodiment 2:
Measure aFGF 2～30,000 AU, hyaluronic acid 1～6g, mannitol 3～8g, dextran 3～88g, water-solubility collagen 1～6g, purified water adds to 100ml, above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Embodiment 3:
Measure aFGF 4～50,000 AU, mannitol 3～8g, dextran 3～8g, water-solubility collagen 1～6g, purified water adds to 100ml, above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Embodiment 4:
Measure aFGF 6～70,000 AU, mannitol 3～8g, dextran 3～8g, hyaluronic acid 1～6g, purified water adds to 100ml, above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Embodiment 5:
Measure aFGF8～90,000 AU, mannitol 3～8g, water-solubility collagen 1～6g, hyaluronic acid 1～6g, purified water adds to 100ml, above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Embodiment 7:
Measure aFGF10 ten thousand AU, mannitol 3～8g, hyaluronic acid 1～6g, purified water adds to 100ml, above-mentioned each composition is dissolved in the cryogenic purincation water, packing, lyophilizing, promptly.
Claims (5)
1, the application of acid fibroblast growth factor in cosmetics.
2, cosmetics according to claim 1 is characterized in that:
Acidic fibroblast stimulating factor 1～100,000 AU
Active stabilizer 4～8g
Accelerative activator 0.3～3g
Protease inhibitor 0.1～0.6g
Percutaneous absorption enhancer 0.1～2g.
3, the described cosmetics of claim 2 is characterized in that: described protease inhibitor is any one in boric acid leucine, boric acid valine, boric acid alanine, the N-acetylcystein.
4, the described cosmetics of claim 2 is characterized in that: described Percutaneous absorption enhancer is any one in laurocapram, propylene glycol, the calcium mercaptoacetate.
5, according to the preparation method of the described cosmetics of claim 1, may further comprise the steps: add in the acid fibroblast growth factor in active stabilizer, accelerative activator, protease inhibitor, the Percutaneous absorption enhancer any one or multiple, be mixed with aqueous solution with purified water, adopt vacuum freeze-drying technique to make lyophilized powder again;
Freeze drying process and condition are: goods are carried out pre-cooling, make it reduce to-60 ℃, temperature fall time is 4-5 hour; Be evacuated to 0.3Mbar; Heat, distillation, the distillation time is 14-18 hour; Water sublimed to goods finishes, and tamponade charges into aseptic gas in the case.
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| CNA2009100669522A CN101548935A (en) | 2009-05-15 | 2009-05-15 | Use of acidic fibroblast growth factor in preparing cosmetics |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109646671A (en) * | 2018-12-26 | 2019-04-19 | 陕西三八妇乐科技股份有限公司 | A kind of growth-factor preparation and its preparation method and application |
-
2009
- 2009-05-15 CN CNA2009100669522A patent/CN101548935A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109646671A (en) * | 2018-12-26 | 2019-04-19 | 陕西三八妇乐科技股份有限公司 | A kind of growth-factor preparation and its preparation method and application |
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