CN104606667B - BFGF bovine basic fibroblast growth factor gel - Google Patents
BFGF bovine basic fibroblast growth factor gel Download PDFInfo
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- CN104606667B CN104606667B CN201510074110.7A CN201510074110A CN104606667B CN 104606667 B CN104606667 B CN 104606667B CN 201510074110 A CN201510074110 A CN 201510074110A CN 104606667 B CN104606667 B CN 104606667B
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Abstract
The present invention relates to field of biological product, particularly a kind of bFGF bovine basic fibroblast growth factor gel and preparation method thereof.The component of described gel and consumption are: bFGF bovine basic fibroblast growth factor 20000-22000IU, human albumin 0.8-1.5mg, carbomer 20-30mg, glycerol 100-120mg, and pH adjusting agent is appropriate, and water for injection adds to 5ml.Gel-type vehicle compatibility in inventive gel agent is reasonable, and easily coating, ductility are good, fine and smooth evenly, non-stimulated to skin, moistening effect is good and be easy to skin absorption; And this gel stability is high, lasting medicine, quality is secure, can wound healing more effectively, repairs trauma skin; Be conducive to the promotion and application of this bFGF bovine basic fibroblast growth factor gel.
Description
Technical field
The present invention relates to field of biological product, particularly relate to a kind of bFGF bovine basic fibroblast growth factor gel and preparation method thereof.
Background technology
Skin is the maximum organ of people's bulk area, because it has stop that foreign body and pathogen invade, prevents the functions such as body fluid loss, thus plays important barrier protection effect.Skin also has sensation, protection, reconciles the important physiological function such as body temperature, and to maintain opposition and the unification of body and external environment, skin is again the important component part of body immune system simultaneously.Any reason causes the destroyed and certainty damage of skin seriality, if repair not in time, then can produce the acute and chronic infection of wound surface and corresponding complication thereof, causes tremendous influence to the body & mind of patient.
Traditional skin wound reparation generally adopts the methods such as autograft, allograft, using artificial substitute and organization engineering skin.Although autologous tissue or organ transplantation have good repairing and treating effect, face the restriction that donor source lacks, needs the conditions such as second operation; Allosome, xenotransplantation not only face the few restriction of donor source, and existence causes immunological rejection, propagates cause of disease equivalent risk; Artificial substituent then only can play reparation or the filling effect of limited tissue or organ; Although organization engineering skin has easy acquisition, external can the holding time long, there is the advantage of the barrier function preventing antibacterial intrusion and loss of moist, but the multiplication capacity due to the human tissue cell of original cuiture is limited and zooblast may cause the problems such as immunological rejection, is difficult to meet clinical demand.Therefore, a kind of method working out effective reparation skin trauma is the clinical difficult problem needing solution badly.
Basic fibroblast growth factor (Basicfibroblastgrowthfactor, be abbreviated as bFGF) be also called corium somatomedin, in the keratinocyte of main existence and epidermal basal, it is important short cell mitogen Summing Factor angiogenesis factor, to deriving from mesoderm and neuroectodermal polytype cell has biologic activity widely, cell proliferation can be promoted, growth, differentiation, promote angiogenesis, improving cell life is environment, promote injured nerve regeneration and blood vessel, muscle, the reparations such as skin, reduce the infringement that ultraviolet radiation causes skin, neurotrophic and regeneration factor.A large amount of basic research and clinical trial confirm, bFGF can act on the different aspects of skin, promotes cell regeneration comprehensively, increase the immunologic function of skin self, improve circulation, promote skin metabolism, make skin reach best states of physiologic function, repair damaged skin.
BFGF bovine basic fibroblast growth factor (Recombinantbovinebasicfibroblastgrowthfactor, be abbreviated as rb-bFGF) be by the escherichia coli containing high expression human/bovine basic fibroblast growth factor gene, the active factors obtained after fermenting, be separated and be highly purified.Find after deliberation, rb-bFGF can wound healing, for wound skin such as empyrosis wound surface, chronic wound and fresh wound surface.
Chinese patent CN102357242B discloses a kind of bFGF bovine basic fibroblast growth factor gel, the gel-type vehicle of this somatomedin gel is hydroxypropyl emthylcellulose and carbomer, this gel good stability and the good advantage of release property, but, slight opalescence is had after hydroxypropyl emthylcellulose in the gel-type vehicle of this gel is water-soluble, and dissolubility is less in the hot water, easy precipitation, gel-type vehicle containing hydroxypropyl emthylcellulose is not suitable for high temperature sterilize, certain safety is there is in this gel to wound skin, but also the ductility of this gel can be affected, be unfavorable for that patient uses.
Summary of the invention
The technical problem to be solved in the present invention is lower to the safety of existing bFGF gel, the defect of poor ductility, object of the present invention provides a kind of good stability, nonirritant, moistening effect good and easily absorb bFGF bovine basic fibroblast growth factor gel and preparation method thereof.
Goal of the invention of the present invention is realized by following embodiment.
BFGF bovine basic fibroblast growth factor gel of the present invention, the component of described gel and consumption are:
BFGF bovine basic fibroblast growth factor 20000-22000IU;
Human albumin 0.8-1.5mg;
Carbomer 20-30mg;
Glycerol 100-120mg;
PH adjusting agent is appropriate;
Water for injection adds to 5ml;
Wherein, described pH adjusting agent is sodium hydroxide or triethanolamine.
BFGF bovine basic fibroblast growth factor gel of the present invention, the component of described gel and consumption are preferably:
BFGF bovine basic fibroblast growth factor 21000IU;
Human albumin 1mg;
Carbomer 25mg;
Glycerol 110mg;
PH adjusting agent is appropriate;
Water for injection adds to 5ml;
Wherein, described pH adjusting agent is sodium hydroxide or triethanolamine.
In addition, the present invention also provides a kind of preparation method of bFGF bovine basic fibroblast growth factor gel, comprises following steps:
(1) water for injection is poured in stainless steel cask, under stirring, add carbomer and the glycerol of recipe quantity, swellingly to spend the night, obtain carbomer solution for subsequent use;
(2) pH adjusting agent of recipe quantity is added the carbomer solution obtained in step (1), adjust ph is stir after 6.5-7.5, pours dilute preparing tank into;
(3) add appropriate injection pond and wash stainless steel cask, swing the injection water after washing and pour dilute preparing tank in the lump, sterilizing 30min under 121 DEG C of hot pressing, be cooled to 37 DEG C clear gel for subsequent use;
(4) recipe quantity bFGF bovine basic fibroblast growth factor and human albumin are dissolved in appropriate water for injection, degerming with 0.22 μm of membrane filtration;
(5) the solution mix homogeneously that the clear gel aseptically step (3) obtained and step (4) obtain, subpackage and get final product.
Carbomer substrate of the present invention is a kind of aqueous gel substrate, the effect of low tempertaure storage and high-temperature heat sterilization can be tolerated, ductility is good, be easy to smear, and have good biocompatibility, eyes and skin are not stimulated, simultaneously, the carbomer substrate of adding glycerol can promote the absorption of skin, greatly increase its moistening effect, moistening environment is more conducive to the healing of wound, prove that bFGF bovine basic fibroblast growth factor gel of the present invention is that ductility is high from the ductility of embodiments of the invention 7 and moisture retention test, the gel that moistening effect is high, promote reparation and the healing of the skin person of being wound, be beneficial to the promotion and application of this gel bFGF bovine basic fibroblast growth factor gel.
The present invention is drawn by the wound tissue healing test of test example 8 pairs of Cavia porcelluss, bFGF bovine basic fibroblast growth factor gel of the present invention obviously can reduce the time of the incrustation of guinea pig back scalded skin and healing, promote the healing of scalded skin, reduce healing time, illustrate that bFGF bovine basic fibroblast growth factor gel of the present invention has remarkable facilitation to wound tissue healing.
The present invention can be drawn by the clinical trial of test example 9, and from I degree of burn and scald therapeutic effect, the cure rate for the treatment of 1 group is 100%, 16 equal preventing from scar of clinical volunteers; Light degree Ⅱ burn and scald therapeutic effect is seen, the cure rate for the treatment of 4 groups is 92.14%, owing to treating the cure rate of 5 groups and treatment 6 groups, and the cicatrix rate for the treatment of 4 groups is only 3.86%, with treatment 5 groups with treat 6 groups and compare, significantly reduce its cicatrix rate, prove that bFGF bovine basic fibroblast growth factor gel of the present invention achieves unforeseeable technique effect at raising cure rate and staying in scar problem of burnt degree.
In a word, the present invention compared with prior art, has following advantage and effect:
1. the gel-type vehicle compatibility in inventive gel agent is reasonable, and easily coating, ductility are good, fine and smooth evenly, non-stimulated to skin, good and the easy and skin of moistening effect absorbs;
2. gel stability of the present invention is high, lasting medicine, and quality is secure, can wound healing more effectively, shortens scab forming time, reduces cicatrix rate;
3. inventive gel agent is easy to use, economical and practical, adds the compliance of patient.
Detailed description of the invention
Be described further below in conjunction with preferred embodiment of the present invention, these embodiments are interpreted as limiting practical range of the present invention, and its modification and equivalent way all covered in protection scope of the present invention.
The preparation of embodiment 1, bFGF bovine basic fibroblast growth factor gel of the present invention
Prescription:
BFGF bovine basic fibroblast growth factor 21000000IU;
Human albumin 1.0g;
Carbomer 25g;
Glycerol 110g;
Triethanolamine is appropriate;
Water for injection adds to 5000ml;
Make 1000 altogether
Preparation technology is as follows:
(1) water for injection is poured in stainless steel cask, under stirring, add carbomer and the glycerol of recipe quantity, swellingly to spend the night, obtain carbomer solution for subsequent use;
(2) triethanolamine of recipe quantity is added the carbomer solution obtained in step (1), adjust ph is stir after 6.5-7.5, pours dilute preparing tank into;
(3) add appropriate injection pond and wash stainless steel cask, swing the injection water after washing and pour dilute preparing tank in the lump, sterilizing 30min under 121 DEG C of hot pressing, be cooled to 37 DEG C clear gel for subsequent use;
(4) recipe quantity bFGF bovine basic fibroblast growth factor and human albumin are dissolved in appropriate water for injection, degerming with 0.22 μm of membrane filtration;
(5) the solution mix homogeneously that the clear gel aseptically step (3) obtained and step (4) obtain, subpackage and get final product.
The preparation of embodiment 2, bFGF bovine basic fibroblast growth factor gel of the present invention
Prescription:
BFGF bovine basic fibroblast growth factor 20000000IU;
Human albumin 0.8g;
Carbomer 20g;
Glycerol 100g;
Sodium hydroxide q. s;
Water for injection adds to 5000g;
Make 1000 altogether
Preparation technology is as embodiment 1.
The preparation of embodiment 3, bFGF bovine basic fibroblast growth factor gel of the present invention
Prescription:
BFGF bovine basic fibroblast growth factor 22000000IU;
Human albumin 1.5g;
Carbomer 30g;
Glycerol 120g;
Sodium hydroxide q. s
Water for injection adds to 5000g;
Make 1000 altogether
Preparation technology is as embodiment 1.
The preparation of comparative example 4, a kind of bFGF bovine basic fibroblast growth factor gel
Prescription:
BFGF bovine basic fibroblast growth factor 28000000IU;
Human albumin 2.4g;
Hydroxypropyl emthylcellulose 10g
Sodium hydroxide q. s
Water for injection adds to 5000g;
Make 1000 altogether
Preparation technology is as embodiment 1.
The preparation of comparative example 5, a kind of bFGF bovine basic fibroblast growth factor gel
Prescription:
BFGF bovine basic fibroblast growth factor 18000000IU;
Human albumin 2.0g;
Carbomer 25g;
Sodium hydroxide q. s;
Water for injection adds to 5000g;
Make 1000 altogether
Preparation technology is as embodiment 1.
Test example 6, bFGF bovine basic fibroblast growth factor gel long-term stable experiment of the present invention
BFGF bovine basic fibroblast growth factor gel fill embodiment 1 prepared is in undercoating sealing aluminum pipe, test at sealing is placed in 2 DEG C ~ 8 DEG C, respectively at 0th month, 1 month, 3 months, 6 months, 9 months, 12 the end of month, 18 outward appearances detecting gel the end of month, loading amount, pH value, biologic activity (%) and sterility tests.Result is as following table:
Table 1, bFGF bovine basic fibroblast growth factor gel stability of the present invention are tested
Drawn by table 1 data, bFGF bovine basic fibroblast growth factor gel of the present invention, at 2 DEG C ~ 8 DEG C, indices conformance with standard in 18 months, illustrate that bFGF bovine basic fibroblast growth factor gel stability of the present invention is high, be beneficial to suitability for industrialized production and the application of this bFGF bovine basic fibroblast growth factor gel.
The ductility of test example 7, bFGF bovine basic fibroblast growth factor gel and moisture retention test
1, test material
1.1, medication is tested: somatomedin gel prepared by the embodiment of the present invention 1, somatomedin gel prepared by comparative example 4 and somatomedin gel prepared by comparative example 5.
1.2, experimenter: leave cicatrix after filtering out burn and scald healing, 50 volunteers without dermatosis allergies test.
2, test method
2.1, the test method of ductility: 0.02mL gel (only have the grain of rice large) extruded by bFGF bovine basic fibroblast growth factor gel embodiment 1, comparative example 4 and comparative example 5 prepared at 0 scale place of backing plate, use spreading rod pulls, and measures it and pulls length.
2.2, the test method of moisture retention: the initial water score value leaving cicatrix position after first adopting moisture detector to test the burn-healing of 50 volunteers, then cicatrix position is left after bFGF bovine basic fibroblast growth factor gel prepared by embodiment 1, comparative example 4 and comparative example 5 being spread upon burn-healing, the difference size analyzing moisture content value evaluates the moisture retention of somatomedin gel, the moisture value percentage rate wherein after skin moisture value climbing=coating-initial water score value percentage rate.
3, result of the test
Result of the test is as shown in table 2, pull length be 50 volunteers on average pull length, skin moisture value climbing is the average skin moisture value climbing of 50 volunteers.
The ductility of table 2 bFGF bovine basic fibroblast growth factor gel and moisture retention result of the test
As shown in Table 2, the ductility of bFGF bovine basic fibroblast growth factor gel of the present invention and moisture retention are all better than comparative example 4 and comparative example 5, illustrate that bFGF bovine basic fibroblast growth factor gel of the present invention is that ductility is high, the gel that moistening effect is high, promote reparation and the healing of the skin person of being wound, be beneficial to the promotion and application of this gel bFGF bovine basic fibroblast growth factor gel.
Test example 8, bFGF bovine basic fibroblast growth factor gel of the present invention are tested the impact of wound tissue healing
1, test material
1.1, experimental animal: Cavia porcellus, male and female half and half, body weight 280-300g;
1.2, trial drug: recombinant bovine basic fibroblast growth somatomedin gel prepared by the embodiment of the present invention 1, control drug: recombinant bovine basic fibroblast growth somatomedin gel prepared by comparative example 4.
2, test method
Modeling: Cavia porcellus is at the equal normal water of duration of test, and full diet, guinea pig back to lose hair or feathers 4cm*4cm by 24h before test.Scald guinea pig back skin with 80 DEG C of thermostatted waters respectively by a glass cylinder, cause II degree scalding model.Be divided into blank group, treatment group and matched group after Cavia porcellus modeling at random, often organize each 10.Treatment group and matched group give relative medicine 4 times every day respectively, raise separately, continuous 7 days.Observe wound healing situation, record skin scab forming time and wound healing time.
3, result of the test
Adopt variance analysis to carry out statistical procedures to result of the test, represent with X ± S, result of the test is as follows:
Table 3 is group Cavia porcellus wound tissue healing results contrast (X ± S) respectively
Compare with blank group: * P<0.05, * * P<0.01; Compare with matched group, #P<0.05.
Shown by above result, bFGF bovine basic fibroblast growth factor gel of the present invention obviously can reduce the time of the incrustation of guinea pig back scalded skin and healing, and healing comparatively matched group is more obvious, illustrate that bFGF bovine basic fibroblast growth factor gel of the present invention has remarkable facilitation to wound tissue healing.
The experimental study of test example 9, bFGF bovine basic fibroblast growth factor gel for treating burn and scald clinical volunteers of the present invention
1, testing data
1.1 physical data
Choose clinical volunteers 102 example of Zhuhai City's burn and scald, man 52 example, female 50 example, 15 ~ 68 years old age, 27 years old mean age; Be divided into according to burn and scald reason: boiling water burn 30 example, flame burn 37 example, steam scalds 30 examples, chemical burn 5 example; Be divided into according to burn and scald degree: I degree of burn and scald 48 example, light degree Ⅱ burn and scald 52 example.
1.2 grouping and medications
I degree of burn and scald clinical volunteers 48 example, be divided into 3 groups, often organize 16 people, concrete medication is as follows:
Treat 1 group: the outer bFGF bovine basic fibroblast growth factor gel being coated with embodiment 1 and preparing;
Treat 2 groups: the outer bFGF bovine basic fibroblast growth factor gel being coated with comparative example 4 and preparing;
Treat 3 groups: the outer bFGF bovine basic fibroblast growth factor gel being coated with comparative example 5 and preparing.
Light degree Ⅱ burn and scald clinical volunteers 54 example, be divided into 3 groups, often organize 18 people, concrete medication is as follows:
Treat 4 groups: the outer bFGF bovine basic fibroblast growth factor gel being coated with embodiment 1 and preparing;
Treat 5 groups: the outer bFGF bovine basic fibroblast growth factor gel being coated with comparative example 4 and preparing;
Treat 6 groups: the outer bFGF bovine basic fibroblast growth factor gel being coated with comparative example 5 and preparing.
2, Therapeutic Method
Under sterile working, by the wound surface routine disinfection of boiling hot burn, first simply clean wound surface with normal saline, remove attachment and the elephant skin that come off, the vesicle tap that tension force is high, keep residual epithelium as far as possible, wipe out and come off or downright bad epidermis, the capable thorough debridement of infective wound surface, be coated with outside wound surface or spray corresponding medicinal liquid, every 3 hours daytimes 1 time, every day coating 4 times.Fully expose wound surface after medication, can not cover, prevent into water, winter is noted warming.
Recovery from illness criterion of therapeutical effect: after medication, swelling disappears, and pain disappears, and wound surface is without infection, and subcrustal healing, has epidermal growth, and wound surface is substantially repaired or all repaired.Record recovery from illness number in 15d after medication, calculate 7d cure rate, recovery from illness number/case total number of persons × 100% in 7d cure rate=7d; Time needed for definition wound surface complete epithelization is healing time, records the healing time of every routine clinical volunteers, and carries out t inspection to healing time and compare; The wound surface cicatrix situation of record burn and scald clinical volunteers, clinical volunteers's number of cases of record scar, calculates cicatrix rate, clinical volunteers's number of cases/case total number of persons × 100% of cicatrix rate=scar; Applied statistics software SPSS11.0 carries out the χ of each treatment group cure rate and cicatrix rate
2inspection.If χ
2> χ
2 0.05,1=3.84, with p<0.05, there is significant difference.The results are shown in Table 4.
3, result of the test
Table 4 bFGF bovine basic fibroblast growth factor gel is to the clinical treatment situation of burn and scald clinical volunteers
As shown in Table 4, from I degree of burn and scald therapeutic effect, the cure rate for the treatment of 1 group is 100%, 16 equal preventing from scar of clinical volunteers; Light degree Ⅱ burn and scald therapeutic effect is seen, the cure rate for the treatment of 4 groups is 92.14%, and owing to treating 5 groups and the treatment cure rate of 6 groups, and the cicatrix rate for the treatment of 4 groups is only 3.86%, with treatment 5 groups with treat 6 groups and compare, significantly reduce its cicatrix rate.Visible, bFGF bovine basic fibroblast growth factor gel of the present invention achieves unforeseeable technique effect at raising cure rate and staying in scar problem of burnt degree.
Claims (2)
1. a bFGF bovine basic fibroblast growth factor gel, is characterized in that, the component of described gel and consumption are:
BFGF bovine basic fibroblast growth factor 21000IU;
Human albumin 1mg;
Carbomer 25mg;
Glycerol 110mg;
PH adjusting agent is appropriate;
Water for injection adds to 5ml;
Wherein, described pH adjusting agent is sodium hydroxide or triethanolamine.
2. the preparation method of bFGF bovine basic fibroblast growth factor gel according to claim 1, is characterized in that comprising following steps:
(1) water for injection is poured in stainless steel cask, under stirring, add carbomer and the glycerol of recipe quantity, swellingly to spend the night, obtain carbomer solution for subsequent use;
(2) pH adjusting agent of recipe quantity is added the carbomer solution obtained in step (1), adjust ph is stir after 6.5-7.5, pours dilute preparing tank into;
(3) add appropriate injection pond and wash stainless steel cask, swing the injection water after washing and pour dilute preparing tank in the lump, sterilizing 30min under 121 DEG C of hot pressing, be cooled to 37 DEG C clear gel for subsequent use;
(4) recipe quantity bFGF bovine basic fibroblast growth factor and human albumin are dissolved in appropriate water for injection, degerming with 0.22 μm of membrane filtration;
(5) the solution mix homogeneously that the clear gel aseptically step (3) obtained and step (4) obtain, subpackage and get final product.
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| CN109350735A (en) * | 2018-07-18 | 2019-02-19 | 吕向阳 | A kind of preparation for external application to skin and preparation method and application for scar reparation |
| CN113274486A (en) * | 2021-04-15 | 2021-08-20 | 上海腾瑞制药股份有限公司 | Stable acidic fibroblast growth factor preparation and preparation method and application thereof |
| CN115501125A (en) * | 2022-10-20 | 2022-12-23 | 澳门科技大学 | H for repairing skin lesions 2 S donor gel and H 2 Preparation method of S donor gel |
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| CN102357242A (en) * | 2011-11-02 | 2012-02-22 | 珠海亿胜生物制药有限公司 | Recombination cattle alkaline fibroblast growth factor gel |
| CN102389392A (en) * | 2011-11-02 | 2012-03-28 | 珠海亿胜生物制药有限公司 | Recombinant bovine basic fibroblast growth factor gel for eye use |
| CN103083226A (en) * | 2012-10-26 | 2013-05-08 | 温州医学院 | Preparation of gel containing fibroblast growth factor-1 modified body and application thereof for treating diabetic foot |
| CN103768017A (en) * | 2012-10-25 | 2014-05-07 | 复旦大学 | Basic fibroblast growth factor nasal spray for treating Alzheimer's disease |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102357242A (en) * | 2011-11-02 | 2012-02-22 | 珠海亿胜生物制药有限公司 | Recombination cattle alkaline fibroblast growth factor gel |
| CN102389392A (en) * | 2011-11-02 | 2012-03-28 | 珠海亿胜生物制药有限公司 | Recombinant bovine basic fibroblast growth factor gel for eye use |
| CN103768017A (en) * | 2012-10-25 | 2014-05-07 | 复旦大学 | Basic fibroblast growth factor nasal spray for treating Alzheimer's disease |
| CN103083226A (en) * | 2012-10-26 | 2013-05-08 | 温州医学院 | Preparation of gel containing fibroblast growth factor-1 modified body and application thereof for treating diabetic foot |
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Inventor after: Fang Haizhou Inventor after: Dai Peimin Inventor after: Wang Yunhua Inventor after: Jiang Danli Inventor after: Zheng Zanshun Inventor after: Jiang Ruofeng Inventor after: Qiu Qiyu Inventor after: Zhu Aitang Inventor after: Xun Xinping Inventor after: Chen Xionglai Inventor after: Qu Wei Inventor after: Wang Xinzhi Inventor before: Fang Haizhou Inventor before: Zheng Zanshun Inventor before: Jiang Ruofeng Inventor before: Qiu Qiyu Inventor before: Zhu Aitang Inventor before: Lan Xuan Inventor before: Xun Xinping Inventor before: Chen Xionglai |