CN102389392A - Recombinant bovine basic fibroblast growth factor gel for eye use - Google Patents
Recombinant bovine basic fibroblast growth factor gel for eye use Download PDFInfo
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- CN102389392A CN102389392A CN2011103424365A CN201110342436A CN102389392A CN 102389392 A CN102389392 A CN 102389392A CN 2011103424365 A CN2011103424365 A CN 2011103424365A CN 201110342436 A CN201110342436 A CN 201110342436A CN 102389392 A CN102389392 A CN 102389392A
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Abstract
The invention discloses a recombinant bovine basic fibroblast growth factor gel for eye use and a preparation method thereof. The gel for eye use contains the following ingredients with the dosage: 1000 to 3000 IU of recombinant bovine basic fibroblast growth factors, 0.01 to 0.25 mg of a protein protectant, 0.5 to 1.5 mg of a gel matrix, an appropriate amount of a pH regulator, and water for injection added to 0.4 g. The product has the characteristics of having less irritation, being easy to apply and having a long-lasting action.
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 for eye use and preparation method thereof.
Background technology
Basic fibroblast growth factor (Basic fibroblast growth factor; Be abbreviated as bFGF) mainly be distributed in the histoorgan in mesoderm and neural germinal layer source, many histiocytes there is effect, can promote cell division; Regulate cellular metabolism; Influence form, infiltration, differentiation and the aging of cell, the epithelial cell stromal cell of corneal nuclear endotheliocyte all can exert an influence, and the reparation of regeneration, corneal stroma and the endodermis of alkali burn corneal epithelium is all had facilitation.
BFGF bovine basic fibroblast growth factor (Recombinant bovine basic fibroblast growth factor; Be abbreviated as rb-bFGF) be the escherichia coli that efficiently express the human/bovine basic fibroblast growth factor gene by containing, through fermentation, separation and highly purified after the active factors that obtains.Rb-bFGF can treat corneal epithelial defect and point-like keratopathy, recurrent superficial punctate keratopathy, light moderate xerophthalmia, bullous keratitis, corneal abrasion, light moderate chemical burn, the operation on cornea that a variety of causes causes and the postoperative healing is bad, the single phlyctenular corneal ulcer of map shape (or trophism) etc.
Gel for eye use is a kind of gel that is used for eye and since the preparation that is used for eye require fine and smooth even, non-stimulated, be prone to coating etc., therefore, gel for eye use is extremely tight with the consumption requirement to the kind of adjuvant.Simultaneously, in order to obtain persistent drug effect, the selection of gel-type vehicle is also extremely important in the gel for eye use.
One Chinese patent application 200510036465 (the application people is Nanhai Longtime Pharmaceutical Co., Ltd.) discloses a kind of recombination human basic fibroblast growth factor exterior-applied gel that is used to treat diseases such as burn wound; It is made up of recombination human basic fibroblast growth factor stock solution and pharmaceutically acceptable auxiliaries, and wherein pharmaceutically acceptable auxiliaries is selected from Acritamer 940 NF, hyaluronate sodium, trehalose, mannitol, Dextran 40, heparin sodium.One Chinese patent application 200510009218 (the application people is the upright Pharmaceutical Technology Co., Ltd of the two aigrets in Beijing) discloses a kind of recombination human basic fibroblast growth factor exterior-applied gel of treating diseases such as wound, burn and scald, and its gel-type vehicle comprises thickening agent (being selected from sodium carboxymethyl cellulose, methylcellulose, Polyethylene Glycol, carbomer, chitosan that percentage by weight is 0.1-10%), wetting agent, antiseptic, stabilizing agent, pH regulator agent and water.
Though disclosing the gel and the adjuvant thereof of recombination human basic fibroblast growth factor, above-mentioned two patent applications form; But they only are fit to treat wound etc. through the approach of topical administration, and do not do any instruction for how bFGF bovine basic fibroblast growth factor being selected to prepare the gel that is applicable to eye through specific adjuvant.
Therefore, the recombination human basic fibroblast growth factor gel for eye use that needs little, the easy coating of a kind of zest of exploitation, lasting medicine is still arranged.
Summary of the invention
The purpose of this invention is to provide bFGF bovine basic fibroblast growth factor gel for eye use of little, the easy coating of a kind of zest, lasting medicine and preparation method thereof.
Goal of the invention of the present invention realizes through following embodiment.
A kind of bFGF bovine basic fibroblast growth factor gel for eye use is characterized in that the component of said gel for eye use and consumption are:
BFGF bovine basic fibroblast growth factor 1000-3000IU;
Protein protective agent 0.01-0.25mg;
Gel-type vehicle 0.5-1.5mg;
The pH regulator agent is an amount of;
Water for injection adds to 0.4ml;
Wherein, described protein protective agent is selected from human albumin, mannitol and Polyethylene Glycol, preferred human albumin; Described gel-type vehicle is selected from hydroxypropyl emthylcellulose and carbomer; Described pH regulator agent is sodium hydroxide or hydrochloric acid;
One preferred embodiment in, the component of bFGF bovine basic fibroblast growth factor gel for eye use and consumption are:
BFGF bovine basic fibroblast growth factor 1680IU;
Protein protective agent 0.2mg;
Gel-type vehicle 1.00mg;
The pH regulator agent is an amount of;
Water for injection adds to 0.4ml;
Wherein, described protein protective agent is the human albumin; Described gel-type vehicle is hydroxypropyl emthylcellulose and carbomer; Described pH regulator agent is sodium hydroxide or hydrochloric acid;
The method for preparing of bFGF bovine basic fibroblast growth factor gel for eye use of the present invention comprises following steps:
(1) with gel-type vehicle with an amount of water for injection swelling, adjust pH makes the formation clear gel, then at 121 ℃ of pressure sterilizings, cooling back is subsequent use;
(2) bFGF bovine basic fibroblast growth factor and protein protective agent are dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming;
(3) the solution mix homogeneously that clear gel that under aseptic condition, step (1) is obtained and step (2) obtain, packing promptly gets.
Preferably, method for preparing is:
(1) with the 0.5-1.5mg gel-type vehicle with an amount of water for injection swelling, adjust pH makes the formation clear gel, then at 121 ℃ of pressure sterilizings, cooling back is subsequent use;
(2) 1000-3000IU bFGF bovine basic fibroblast growth factor and 0.01-0.25mg protein protective agent are dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming;
(3) the solution mix homogeneously that clear gel that under aseptic condition, step (1) is obtained and step (2) obtain, packing promptly gets.
The present invention has following advantage and effect:
1. gel for eye of the present invention is to suffer from corneal epithelial defect and point-like keratopathy, recurrent superficial punctate keratopathy, light moderate xerophthalmia, bullous keratitis, corneal abrasion, light moderate chemical burn, the operation on cornea that a variety of causes causes and the postoperative healing is bad, the ocular disease patient of the single phlyctenular corneal ulcer of map shape (or trophism) etc. provides a kind of interchangeable administration form;
2. gel for eye of the present invention be prone to coating, fine and smooth evenly, persistent, to the eye nonirritant;
3. gel for eye of the present invention is easy to use, economical and practical, has increased patient's compliance.
The specific embodiment
Do further explanation below in conjunction with preferred embodiment of the present invention, these embodiment are interpreted as can not limiting practical range of the present invention, and its modification and equivalent way all covered in protection scope of the present invention.
Embodiment 1:
Prescription:
Concrete method for preparing is:
With hydroxypropyl emthylcellulose with an amount of water for injection swelling, adjust pH makes the formation clear gel, and then at 121 ℃ of pressure sterilizings, the cooling back is subsequent use; BFGF bovine basic fibroblast growth factor and human albumin are dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming; With above-mentioned clear gel and filtrating mix homogeneously, packing promptly gets under aseptic condition.
Embodiment 2:
Prescription:
Concrete method for preparing is:
With carbomer, hydroxypropyl emthylcellulose with an amount of water for injection swelling, adjust pH makes the formation clear gel, and then at 121 ℃ of pressure sterilizings, the cooling back is subsequent use; BFGF bovine basic fibroblast growth factor, human albumin and mannitol risen be dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming; With above-mentioned clear gel and filtrating mix homogeneously, packing promptly gets under aseptic condition.
Embodiment 3:
Prescription:
Concrete method for preparing is:
With carbomer with an amount of water for injection swelling, adjust pH makes the formation clear gel, and then at 121 ℃ of pressure sterilizings, the cooling back is subsequent use; BFGF bovine basic fibroblast growth factor, mannitol and Polyethylene Glycol are dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming; With above-mentioned clear gel and filtrating mix homogeneously, packing promptly gets under aseptic condition.
Test Example 1:rb-bFGF gel for eye use promotes the experimentation of corneal alkali burn model healing
1, test method
Experimental animal: healthy new zealand white rabbit, body weight 2-3kg, male and female and usefulness, pocket lamp and slit lamp examination anterior chamber of eye no abnormality seen.120 animals are divided into five groups at random, every group of 24 animals.
The rabbit cornea model of alkali burned: the examination of external eye intact animal, normal saline drips washes right eye 2 times, with 0.5% tetracaine eye drop eye drip 2 times; Topical anesthesia, intramuscular injection ketamine (50mg/kg), general anesthesia; Filter paper dick is soaked in 0.5mol/L NaOH solution, remove unnecessary liquid, carefully sticked in the cornea of right eye center district of fixing eyeball position several minutes; Use normal saline flushing cornea and conjunctival sac immediately after taking off filter paper, promptly process the corneal alkali burn model.
Test specimen: the negative control article are not for containing the gel for eye use of bFGF bovine basic fibroblast growth factor, and adjuvant is formed with embodiment 1, are applied to eye injury place every day sooner or later respectively once; Sample of the present invention is embodiment 1 gel for eye use, is applied to eye injury place every day sooner or later respectively once; Positive reference substance is a Solcoseryl Eye-Jel, each one, and four times a day.
Medication: negative control article, gel for eye use of the present invention and positive reference substance respectively to the animal pattern administration, were treated for two weeks continuously.
Evaluation index: respectively treatment 1,2,4,6,10,14 day the time with fluorescent staining slit lamp microscope standard photograph photo, analyze through the computer picture systematic survey and respectively to organize different time corneal epithelium healing area (result sees table 1) and corneal epithelium healing rate (result sees table 2).Corneal epithelium healing area and corneal epithelium healing rate are high more, show that therapeutic effect is good more.
2, result of the test
Table 1, respectively organize the corneal epithelium healing area (mm of different time
2X ± S)
| Treatment time (my god) | Negative control group | Of the present invention group | Positive controls |
| 1 | 12.1±1.5 | 46.1±2.1 | 21.2±4.3 |
| 2 | 39.5±4.1 | 50.2±2.8 | 44.2±2.4 |
| 4 | 31.3±3.9 | 40.7±2.4 | 31.1±4.5 |
| 6 | 32.3±7.2 | 48.5±3.3 | 39.4±2.5 |
| 10 | 37.2±2.4 | 50.7±2.4 | 46.2±4.3 |
| 14 | 35.2±6.5 | 51.2±3.4 | 44.9±4.5 |
Table 2. is respectively organized the corneal epithelium healing rate (% of different time; X ± S)
| Treatment time (my god) | Negative control group | Of the present invention group | Positive controls |
| 1 | 28.3±1.5 | 95.0±1.6 | 41.8±7.3 |
| 2 | 77.1±2.2 | 99.8±0.7 | 87.2±2.5 |
| 4 | 61.1±6.7 | 80.9±6.2 | 61.3±7.4 |
| 6 | 63.1±7.3 | 96.4±1.9 | 77.7±4.3 |
| 10 | 72.7±4.2 | 100±1.1 | 91.1±6.3 |
| 14 | 68.7±8.3 | 100±0.8 | 88.6±3.8 |
Result by table 1 and table 2 can find out that in the time of the 1st day, of the present invention group epithelium healing area is obviously greater than negative control group and positive controls; In the time of the 4th day, the epithelium healing rate appears necessarily repeatedly; When the 10th day and the 14th day, of the present invention group corneal epithelium healing rate is greater than negative control group and positive controls, and with negative control group significant difference arranged.
Test Example 2: stability study
1, high temperature and humidity test:
The gel for eye use fill of embodiment 1 is sealed in the aluminum pipe in undercoating; Sealing places under 60 ℃, relative humidity 75% condition carries out accelerated tests, respectively at the content, pH, outward appearance, stretchability, the aseptic and centrifugal test (300r/min) that detected gel in 1,5,10 day.Result such as following table:
Table 3, hot and humid result
| 1 day | 5 days | 10 days | |
| Content | 99.99％ | 99.98％ | 99.95％ |
| pH | 7.0 | 7.0 | 7.0 |
| Outward appearance | Water white transparency | Water white transparency | Water white transparency |
| Stretchability | Good | Good | Good |
| Aseptic | Qualified | Qualified | Qualified |
| Centrifugal test | No layering | No layering | No layering |
The result shows, gel for eye use of the present invention steady quality in high temperature and humidity test.
2, low-temperature test:
The gel for eye use fill of embodiment 1 is sealed in the aluminum pipe in undercoating, and sealing places under 4 ℃ and experimentizes, respectively at 1,3, detect content, pH, outward appearance, stretchability, the aseptic and centrifugal test (300r/min) of gel June.Result such as following table:
Table 4, low-temperature test result
| 1 month | 3 months | 6 months | |
| Content | 100.01％ | 99.98％ | 99.95％ |
| pH | 7.0 | 7.0 | 7.0 |
| Outward appearance | Water white transparency | Water white transparency | Water white transparency |
| Stretchability | Good | Good | Good |
| Aseptic | Qualified | Qualified | Qualified |
| Centrifugal test | No layering | No layering | No layering |
The result shows, gel for eye use of the present invention steady quality in low-temperature test.
Claims (3)
1. bFGF bovine basic fibroblast growth factor gel for eye use is characterized in that the component of said gel for eye use and consumption are:
BFGF bovine basic fibroblast growth factor 1000-3000IU;
Protein protective agent 0.01-0.25mg;
Gel-type vehicle 0.5-1.5mg;
The pH regulator agent is an amount of;
Water for injection adds to 0.4ml;
Wherein, described protein protective agent is selected from human albumin, mannitol and Polyethylene Glycol, and described gel-type vehicle is selected from hydroxypropyl emthylcellulose and carbomer, and described pH regulator agent is sodium hydroxide or hydrochloric acid;
2. bFGF bovine basic fibroblast growth factor gel for eye use according to claim 1 is characterized in that the component of said gel for eye use and consumption are:
BFGF bovine basic fibroblast growth factor 1680IU;
Protein protective agent 0.2mg;
Gel-type vehicle 1.00mg;
The pH regulator agent is an amount of;
Water for injection adds to 0.4ml;
Wherein, described protein protective agent is the human albumin; Described gel-type vehicle is hydroxypropyl emthylcellulose and carbomer; Described pH regulator agent is sodium hydroxide or hydrochloric acid;
3. the method for preparing of bFGF bovine basic fibroblast growth factor gel for eye use according to claim 1 is characterized in that comprising following steps:
(1) with gel-type vehicle with an amount of water for injection swelling, adjust pH makes the formation clear gel, then at 121 ℃ of pressure sterilizings, cooling back is subsequent use;
(2) bFGF bovine basic fibroblast growth factor and protein protective agent are dissolved in the proper amount of water for injection, with 0.22 μ m membrane filtration degerming;
(3) the solution mix homogeneously that clear gel that under aseptic condition, step (1) is obtained and step (2) obtain, packing promptly gets.
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| CN201110342436A CN102389392B (en) | 2011-11-02 | 2011-11-02 | Recombinant bovine basic fibroblast growth factor gel for eye use |
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| CN201110342436A CN102389392B (en) | 2011-11-02 | 2011-11-02 | Recombinant bovine basic fibroblast growth factor gel for eye use |
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| CN102389392B CN102389392B (en) | 2012-10-24 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104546692A (en) * | 2015-02-12 | 2015-04-29 | 珠海亿胜生物制药有限公司 | Recombinant calf alkaline fibroblast growth factor ophthalmic gel |
| CN104606667A (en) * | 2015-02-12 | 2015-05-13 | 珠海亿胜生物制药有限公司 | Recombinant bovine basic fibroblast growth factor gel |
| CN104984326A (en) * | 2015-07-14 | 2015-10-21 | 珠海亿胜生物制药有限公司 | Recombinant human basic fibroblast growth factor gel without bacteriostatic agent and preparation method thereof |
| JP2020533015A (en) * | 2017-06-23 | 2020-11-19 | 珠海億勝生物製薬有限公司 | A pharmaceutical composition containing recombinant human basic fibroblast growth factor (rh-bFGF) and rh-bFGF. |
| CN113069533A (en) * | 2021-03-08 | 2021-07-06 | 温州医科大学 | Long-acting fibroblast growth factor gel |
| CN115607506A (en) * | 2022-12-17 | 2023-01-17 | 朗肽生物制药股份有限公司 | Preparation method of anhydrous paste containing recombinant human basic fibroblast growth factor |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104546692A (en) * | 2015-02-12 | 2015-04-29 | 珠海亿胜生物制药有限公司 | Recombinant calf alkaline fibroblast growth factor ophthalmic gel |
| CN104606667A (en) * | 2015-02-12 | 2015-05-13 | 珠海亿胜生物制药有限公司 | Recombinant bovine basic fibroblast growth factor gel |
| CN104606667B (en) * | 2015-02-12 | 2016-04-13 | 珠海亿胜生物制药有限公司 | BFGF bovine basic fibroblast growth factor gel |
| CN104984326A (en) * | 2015-07-14 | 2015-10-21 | 珠海亿胜生物制药有限公司 | Recombinant human basic fibroblast growth factor gel without bacteriostatic agent and preparation method thereof |
| JP2020533015A (en) * | 2017-06-23 | 2020-11-19 | 珠海億勝生物製薬有限公司 | A pharmaceutical composition containing recombinant human basic fibroblast growth factor (rh-bFGF) and rh-bFGF. |
| US11254723B2 (en) | 2017-06-23 | 2022-02-22 | Zhuhai Essex Bio-Pharmaceutical Co., Ltd. | Recombinant human-basic fibroblast growth factor (rh-bFGF) and pharmaceutical composition comprising rh-bFGF |
| JP7044870B2 (en) | 2017-06-23 | 2022-03-30 | 珠海億勝生物製薬有限公司 | A pharmaceutical composition containing recombinant human basic fibroblast growth factor (rh-bFGF) and rh-bFGF. |
| CN113069533A (en) * | 2021-03-08 | 2021-07-06 | 温州医科大学 | Long-acting fibroblast growth factor gel |
| CN115607506A (en) * | 2022-12-17 | 2023-01-17 | 朗肽生物制药股份有限公司 | Preparation method of anhydrous paste containing recombinant human basic fibroblast growth factor |
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