CN1895275A - Calf-blood deprotein extract gel - Google Patents
Calf-blood deprotein extract gel Download PDFInfo
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- CN1895275A CN1895275A CNA2005100830412A CN200510083041A CN1895275A CN 1895275 A CN1895275 A CN 1895275A CN A2005100830412 A CNA2005100830412 A CN A2005100830412A CN 200510083041 A CN200510083041 A CN 200510083041A CN 1895275 A CN1895275 A CN 1895275A
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Abstract
A gel of the deproteinized calf blood's extract is prepared proportionally from the liquid extract of deproteinized calf blood, carbomer, the water for injection, sodium hydroxide and bacterial depressant.
Description
Technical field
The present invention relates to a kind of novel form of calf blood protein-removed extraction, particularly calf-blood deprotein extract gel.
Background technology
Present external used medicine dosage form is based on unguentum and liquid dosage form (liniment and eye drop etc.).Liquid dosage form easily trickles, and drug loss is big, and outer time spent fast drying makes effective ingredient lose carrier and difficult the absorption; Much more the substrate of unguentum all be oily matter (seeing with vaseline and glycerol), though unguentum discharges drug slow, also can not trickle, and substrate is difficult for absorbing, and also is difficult for drying, and it is very inconvenient to use.Gel is a substrate with Carmomer etc., and outward appearance is limpid transparent, and is mobile little, organizes easily to absorb, very welcome as the clinical practice dosage form, presents the trend that progressively replaces unguentum.
The calf blood (protein removed) series products can promote picked-up and the utilization of cell to glucose and oxygen.Under situations such as low blood oxygen and energy requirement increase, this product can promote energy metabolism, increases oxygen-supplying amount.Can treat the caused neurologic impairment of brain blood circulation and Dystrophy (ischemic lesions, craniocerebral trauma); The artery vessel disease that can treat last tremulous pulse, venous circulation obstacle slightly and cause, leg ulcer; Also can treat skin burn, scald, erosion, radio-induced skin, mucosa injury; Healing of wound (wound, decubital ulcer).
At present the calf blood (protein removed) series products mainly contains liquid drugs injection, unguentum, tablet, does not have gel, and liquid drugs injection and tablet are the whole body administrations, can not topical application; Unguentum is the topical application dosage form, but the poor permeability of its substrate, local absorption is bad.Gel preparation is the new pharmaceutical dosage form of succeeding in developing in recent years, uses still not general; In calf blood goods field, be engaged in gel preparation exploitation major part at present and still be in the exploratory stage.
Summary of the invention
A kind of novel form that the purpose of this invention is to provide calf blood protein-removed extraction: gel, both can topical application, also can overcome the shortcoming of unguentum poor permeability.
Calf-blood deprotein extract gel provided by the present invention is made by the raw material that comprises following compositions: calf blood (protein removed) extracting solution 20-50 weight portion, carbomer 0.5-1.2 weight portion, water for injection 45-80 weight portion; The quality percentage composition of solid content is 2%-7% in the described calf blood (protein removed) extracting solution.
In order to regulate the pH value of substrate, described raw material also comprises the sodium hydroxide of 0.2-0.5 weight portion.
Described raw material also comprises the antibacterial of 0.05-0.1 weight portion.
Described antibacterial can be nipagin A, dimethyl fumarate or low-grade fatty acid ester.
Described nipagin A can be ethylparaben; Described low-grade fatty acid ester can be monoglyceride.
Described calf blood (protein removed) extracting solution can be ZL 96120777.9 according to the patent No., the method preparation of denomination of invention for describing in " a kind of method of extracting calf blood protein-removed extraction ": adopt 1-6 monthly age children cattle venous blood, isolate serum, use the ethanol Deproteinization, ethanol is removed in decompression, adds the compound protein enzyme hydrolysis in remaining liquid, and supernatant is stayed in centrifugalize, described supernatant is held back with 5000 dalton's hollow fiber column ultrafilter, collects the filtrate that obtains and is the calf blood (protein removed) extracting solution; Wherein said during with the ethanol Deproteinization consumption of ethanol be that the quality percentage composition that doubles the serum volume is 96% ethanol; Wherein said decompression is removed ethanol for utilizing the rotary evaporation in vacuo instrument, and 37 ℃, 1 atmospheric pressure decompression removes ethanol; Wherein said adding compound protease is hydrolyzed to and adds compound protein enzyme hydrolysis 24 hours; Centrifugalize is 10000rpm, 4 ℃ after the enzyme hydrolysis of wherein said adding compound protein, centrifugal 30 minutes, stays supernatant.
Described carbomer can be carbomer 934 GE, 910,934,934P, 940,941,1342 etc.
Calf-blood deprotein extract gel of the present invention is substrate with the carbomer, and release is fast, no greasy, easily be coated with exhibition, the easy infiltration; Sodium hydroxide and carbomer be proportioning according to a certain percentage, and the pH value that can regulate substrate guarantees the comfortableness that gel uses to optimum; A certain amount of antibacterial can guarantee the stability of exterior-applied formulation.The calf blood (protein removed) gel has remedied the defective that injection and tablet can not topical application, has also overcome local deficiency with the unguentum poor permeability.Calf-blood deprotein extract gel of the present invention can be used for treating traumatic, struvite and trophism cornea, conjunctive disorder; Xerophthalmia; The initial therapy of the ulcer that a variety of causes causes (as various foots ulcer, x ray ulcer, decubitus ulcer); The blood circulation serious hindrance that arteriosclerosis and/or diabetes cause and the dystrophic infringement that causes and burn, skin burn.Experimental result shows that calf-blood deprotein extract gel of the present invention is used for local patholoic changes such as cornea ulcer and burn, and evident in efficacy, its outward appearance is limpid transparent, and viscosity is moderate, is applied to eye and can cause obstacle to eye function.
The specific embodiment
Experimental technique among the following embodiment if no special instructions, is conventional method.
Percentage composition among the following embodiment if no special instructions, is the quality percentage composition.
Embodiment 1, production calf blood (protein removed) extracting solution content are 20% calf-blood deprotein extract gel
Produce 1000 calf-blood deprotein extract gels (every 20g contains calf blood (protein removed) extracting solution 20%, carbomer 0.8%, ethylparaben 0.1%).
The raw material of this calf-blood deprotein extract gel is composed as follows:
Calf blood (protein removed) extracting solution 4000g
Carbomer (934) 160g
Sodium hydroxide 64g
Ethylparaben 20g
Water for injection 15756g
Above-mentioned raw materials is made 1000 calf-blood deprotein extract gels (every 20g contains 20% calf blood protein-removed extraction), and concrete grammar is as follows:
1, preparation calf blood (protein removed) extracting solution
Adopt the venous blood of the purebred cattle in northeast at 1-6 monthly age under the aseptic condition, under the aseptic condition, 4000rpm, 4 ℃, centrifugal 15 minutes separation of serum, get 20L serum, add 40L 96% ethanol, constantly stir, left standstill 60 minutes, make protein denaturation precipitation, through 4 ℃ of centrifugal 30min of 20000rpm, abandon precipitation then, obtain the 50L supernatant, utilize the rotary evaporation in vacuo instrument, 37 ℃, 1 atmospheric pressure decompression removes ethanol, in remaining liquid, add the 6g compound protease, 37 ℃ of hydrolysis 24 hours, at 10000rpm, 4 ℃, centrifugal 30 minutes, stay supernatant, supernatant is held back with 5000 dalton's hollow fiber column ultrafilter, collects the filtrate that obtains and is the calf blood (protein removed) extracting solution, obtains 4000ml (g) calf blood (protein removed) extracting solution.
2, preparation calf-blood deprotein extract gel
(1) utensil is prepared: all utensils (stirred tank, sealing tucker) are cleaned, sterilized.
(2) substrate preparation: take by weighing carbomer (934) 160g, ethylparaben 20g adds injection water 15756g, sodium hydroxide 64g, mixing in stirred tank.
(3) substrate sterilization: with substrate under 121 ℃ of conditions, autoclaving 20min.
(4) half-finished preparation: get calf blood (protein removed) extracting solution 4000g, add in the aseptic substrate, abundant mixing, being adjusted to pH value with 0.1mol/L NaOH or 0.1mol/L HCl is 5.5～8.5.
(5) packing: install in the aluminum pipe every 20g with sealing ointment tucker branch.
(6) check, packing: the quality standard by calf blood (protein removed) ointment finished product is tested, and eligible is packaged into finished product.
Embodiment 2, production calf blood (protein removed) extracting solution content are 50% calf-blood deprotein extract gel
Produce 1000 calf blood (protein removed) gels (every 5g contains calf blood (protein removed) extracting solution 50%, carbomer (934) 1%, the ratio of dimethyl fumarate is 0.05%)
The raw material of this calf-blood deprotein extract gel is composed as follows:
Calf blood (protein removed) extracting solution 2500g
Carbomer (934) 50g
Sodium hydroxide 20g
Dimethyl fumarate 2.5g
Water for injection 2427.5g
Above-mentioned raw materials is made 1000 calf-blood deprotein extract gels (every 5g contains 50% calf blood (protein removed) extracting solution), and concrete grammar is as follows:
1, preparation calf blood (protein removed) extracting solution
Adopt the venous blood of the purebred cattle in northeast at 1-6 monthly age under the aseptic condition, under the aseptic condition, 4000rpm, 4 ℃, centrifugal 15 minutes separation of serum, get 12L serum, add 24L 96% ethanol, constantly stir, left standstill 60 minutes, make protein denaturation precipitation, through 4 ℃ of centrifugal 30min of 20000rpm, abandon precipitation then, obtain the 30L supernatant, utilize the rotary evaporation in vacuo instrument, 37 ℃, 1 atmospheric pressure decompression removes ethanol, in remaining liquid, add the 3.6g compound protease, 37 ℃ of hydrolysis 24 hours, at 10000rpm, 4 ℃, centrifugal 30 minutes, stay supernatant, supernatant is held back with 5000 dalton's hollow fiber column ultrafilter, collects the filtrate that obtains and is the calf blood (protein removed) extracting solution, obtains 2500g calf blood (protein removed) extracting solution.
2, preparation calf-blood deprotein extract gel
(1) utensil is prepared: all utensils (stirred tank, sealing tucker) are cleaned, sterilized.
(2) substrate preparation: take by weighing carbomer (934) 50g, dimethyl fumarate 2.5g adds injection water 2427.5g, sodium hydroxide 20g, mixing.
(3) substrate sterilization: with substrate under 121 ℃ of conditions, autoclaving 20min.
(4) half-finished preparation: get calf blood (protein removed) extracting solution 2500g, add in the aseptic substrate, abundant mixing, being adjusted to pH value with 0.1mol/L NaOH or 0.1mol/L HCl is 5.5～8.5.
(5) packing: install in the aluminum pipe every 5g with sealing ointment tucker branch.
(6) check, packing: the quality standard by calf blood (protein removed) ointment finished product is tested, and eligible is packaged into finished product.
Embodiment 3, calf-blood deprotein extract gel are to the influence of rabbit corneal alkali burn epithelial repair
Experimental drug:
Calf blood (protein removed) extracting solution content is 20%, 50%, 70% calf blood (protein removed) extracting solution gel, and specification is 5 and restrains/prop up.Their production technology is with embodiment 1, just raw material is formed different, calf blood (protein removed) extracting solution content is that the raw material of 20% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 1g/ and props up, carbomer (934) 0.04g/ props up, sodium hydroxide 0.016g/ props up, ethylparaben 0.005g/ props up, and water for injection 3.939g/ props up; Calf blood (protein removed) extracting solution content is that the raw material of 50% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 2.5g/ and props up, carbomer (934GE) 0.05g/ props up, sodium hydroxide 0.02g/ props up, and ethylparaben 0.0025g/ props up, and water for injection 2.4275g/ props up; Calf blood (protein removed) extracting solution content is that the raw material of 70% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 3.5g/ and props up, carbomer (934GE) 0.05g/ props up, sodium hydroxide 0.02g/ props up, and ethylparaben 0.0025g/ props up, and water for injection 1.4275g/ props up.
Solcoseryl Eye-Jel (young Sanguis Bovis seu Bubali extract eye ointment): Switzerland Ba Sai and plain tall and big pharmaceutical factory produces, specification is 20% (calf blood (protein removed) extracting solution content), 5 grams/.
Japanese white big ear rabbit is adopted in this experiment, and (Jinzhou Medical College's Experimental Animal Center, the quality certification number: No. 038, distant real kinoplaszm card word) 40, body weight is 2.5～3.5Kg, male and female dual-purpose, healthy no oculopathy.With 1% tetracaine topical anesthesia.With the diameter that is soaked with 0.5mol/L NaOH solution is the filter paper of 8mm, puts anterior corneal surface central authorities, keeps 1min and takes off, and washes 2min continuously with sterile saline, makes model of alkali burned.Behind every rabbit right side corneal alkali burn, animal is divided into 5 groups at random, i.e. three experimental grouies of calf-blood deprotein extract gel (II, III, IV group), Solcoseryl Eye-Jel experimental group (V group), PBS buffer blank group (I group).
I group: eye drop solvent (PBS buffer)
II group: 20% calf-blood deprotein extract gel
III group: 50% calf-blood deprotein extract gel
IV group: 70% calf-blood deprotein extract gel
V group: Solcoseryl Eye-Jel
Medication: each 1, every day 4 times, continuous 2 weeks.Carry out the scoring of cornea clinical sign and calculate the corneal epithelium healing rate.
1, cornea sign scoring:
All animals were respectively at the 0th, 1,2,3,5,7, the 10 and 14 day cornea sign with each group of slit lamp examination.Standards of grading with reference to Ando etc. are evaluated:
1. the corneal opacity
0 minute: corneal transparency, do not have muddy;
1 minute: nubecula, iris texture were as seen;
2 minutes: cornea moderate muddiness, iris texture is unclear;
3 minutes: cornea moderate muddiness, conceal and see pupil;
4 minutes: cornea severe muddiness, pupil loseed.
2. epithelium fluorescent staining
0 minute: the cornea non-coloring;
1 minute: corneal dyeing area≤1/4 quadrant;
2 minutes: corneal dyeing area＞1/4 but≤1/2 quadrant;
3 minutes: corneal dyeing area＞1/2 but≤3/4 quadrant;
4 minutes: corneal dyeing area 〉=3/4 quadrant.
All have any performance and all keep the score, then the obatained score addition is clinical cumulative point (clinical accumulation score, CAS).
Cornea clinical sign appraisal result is as shown in table 1.
Table 1 test group and the clinical cumulative point of matched group cornea (X ± SD, n=8)
| Group | 0 day | The 1st day | The 2nd day | The 3rd day | The 5th day | The 7th day | The 10th day | The 14th day |
| I II III IV V | 8.0±0 8.0±0 8.0±0 8.0±0 8.0±0 | 7.67±0.58 7.76±0.58 7.33±0.58 7.30±0.00 7.68±0.58 | 6.33±0.58 5.68±0.58 6.00±0.00 5.67±0.58 6.00±0.00 | 5.00±0.00 4.33±0.58 4.33±0.58 4.00±0.00 4.21±0.00 | 6.33±0.58 5.33±0.58 a 5.00±0.00 a 5.33±0.58 a 5.25±0.00 a | 6.00±0.00 4.68±0.58 a 4.33±0.58 a 4.33±0.58 a 4.67±0.58 a | 5.68±0.58 3.68±0.58 a 3.00±0.00 a，b，c 2.98±0.58 a，b，c 3.60±0.00 a | 4.33±0.58 3.33±0.58 a 2.33±0.577 a，b，c 2.33±0.58 a，b，c 3.33±0.58 a |
Annotate: a compares P＜0.05:b and compares P＜0.05:c with the II group and compare P＜0.05 with the V group with the I group
2, corneal epithelium healing:
Adopt fluorescent staining method: with one of 20% fluorescein sodium sterile solution, splash in the rabbit conjunctival sac, make its passive 5min of closing one's eyes, stop a moment then slightly, if ophthalmic still has more dyestuff, available normal saline solution flushing is removed it, uses slit lamp observation then.
Carried out slit lamp examination in 0,1,2,3,5,7,10,14 day respectively at hindering the back, fluorescent staining is taken a picture, and photo machine image analysis system is as calculated handled, and calculates its corneal epithelium healing rate.
Date processing: experiment gained data are represented with means standard deviation, with t check carrying out statistical analysis.
Corneal epithelial wound area such as table 2, epithelium healing rate such as table 3.
Table 2 corneal epithelial wound area (mm
2, X ± SD)
| Group | 0 day | The 1st day | The 2nd day | The 3rd day | The 5th day | The 7th day | The 10th day | The 14th day |
| I II III IV V | 50.12±0.15 50.12±0.16 50.26±0.20 49.93±0.26 50.58±0.35 | 36.62±1.42 35.98±2.02 34.45±1.74 35.73±2.37 37.82±1.35 | 36.48±1.87 35.82±1.85 34.15±2.3 35.71±2.5 37.70±0.42 | 27.57±3.32 30.92±1.46 30.44±1.59 28.77±1.25 28.16±1.10 | 39.47±3.5 35.52±2.3 a 33.71±2.18 abc 33.29±1.6 ab 37.29±1.08 ab | 39.17±3.21 28.21±5.21 a 25.78±3.61 abc 25.68±3.81 ab 27.26±3.50 ab | 32.5±2.5 21.8±2.73 a 18.11±2.07 abc 17.95±1.34 ab 20.25±1.42 ab | 28.67±1.91 20.7±0.93 a 16.8±0.33 abc 16.67±0.44 abc 20.23±0.36 ab |
Annotate: a compares P＜0.05:b and compares P＜0.05 with the II group with the I group; C compares P＜0.05 with the V group
Table 3 corneal epithelium healing rate (%)
| Group | 0 day | The 1st day | The 2nd day | The 3rd day | The 5th day | The 7th day | The 10th day | The 14th day |
| I II III IV V | 26.9 28.2 27.5 28.4 28.2 | 27.2 28.5 28.1 28.5 28.5 | 45 38.3 42.4 39.4 39.3 | 21.2 29.1 32.9 29.3 28.3 | 21.8 43.7 48.7 48.5 43.1 | 35.2 56.6 64 64 57.9 | 42.8 58.8 66.6 66.7 59.9 |
The result shows that the basic, normal, high dosage group of calf-blood deprotein extract gel and each data cornea cumulative point of Solcoseryl Eye-Jel group and corneal epithelial wound area have remarkable decline, and comparing with the blank group all has significant difference P＜0.05; The calf-blood deprotein extract gel effect of middle and high dosage group is apparently higher than low dose group and Solcoseryl Eye-Jel group, and there was no significant difference between the middle and high dosage group calf-blood deprotein extract gel.Table it can also be seen that the effect of medicine prolongation in time acts on more obvious thus.
The main component of calf-blood deprotein extract gel is a calf blood protein-removed extraction, and it is made up of small organic molecules such as inorganic ions and aminoacid, small-molecular peptides, nucleotide, oligosaccharide, lipids.These small-molecule substances that calf blood protein-removed extraction comprised can improve picked-up and the utilization of histiocyte to oxygen and glucose, can also promote the migration and the propagation of fibroblast and vascular endothelial cell, thereby can promote wound healing.
Low dosage calf-blood deprotein extract gel and matched group and Solcoseryl Eye-Jel group relatively, calf-blood deprotein extract gel group epithelium healing rate height is significantly higher than matched group, with more but there was no significant difference of Solcoseryl Eye-Jel group; The corneal epithelium damage phenomenon that comes off again appearred on the 3rd day, but relatively degree of injury is lighter for calf-blood deprotein extract gel group and Solcoseryl Eye-Jel group and matched group, the scope that comes off is little, and middle and high dosage group prolongation in time, the corneal epithelium healing rate is all above 65%, be significantly higher than low dose group and Solcoseryl Eye-Jel group, and the epithelium healing rate of matched group only 42.8%, and healing rate is slow.
The calf-blood deprotein extract gel group can significantly promote the reparation of corneal epithelium, improves the immediate union rate, delay to a certain extent or alleviate the corneal epithelium damage that comes off again, and middle concentration curative effect is preferable.
The influence that embodiment 4, calf-blood deprotein extract gel heal to the rat diabetes skin ulcer
Experimental drug:
Calf blood (protein removed) extracting solution content is 10%, 20%, 30% calf-blood deprotein extract gel, and specification is 20 and restrains/prop up.Their production technology is with embodiment 1, just raw material is formed different, calf blood (protein removed) extracting solution content is that the raw material of 10% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 2g/ and props up, carbomer (934GE) 0.16g/ props up, sodium hydroxide 0.064g/ props up, ethylparaben 0.02g/ props up, and water for injection 17.756g/ props up; Calf blood (protein removed) extracting solution content is that the raw material of 20% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 4g/ and props up, carbomer (934GE) 0.16g/ props up, sodium hydroxide 0.064g/ props up, and ethylparaben 0.02g/ props up, and water for injection 15.756g/ props up; Calf blood (protein removed) extracting solution content is that the raw material of 30% calf-blood deprotein extract gel consists of calf blood (protein removed) extracting solution 6g/ and props up, carbomer (934GE) 0.16g/ props up, sodium hydroxide 0.064g/ props up, and ethylparaben 0.02g/ props up, and water for injection 13.756g/ props up.
Solcoseryl Eye-Jel (calf blood protein-removed extraction ointment): Switzerland Ba Sai and plain tall and big pharmaceutical factory produces, specification is 10% (calf blood (protein removed) extracting solution content), 20 grams/.
1, preparation diabetes wound surface model
Get male regular grade SD rats of 3 monthly ages (Jinzhou Medical College's Experimental Animal Center), 12h fasting before the body weight 200g-250g experiment, quantitatively drinking-water.Experiment was weighed the same day, tail vein blood and collection urine, with blood glucose meter (One touch profile blood glucose meter: Lifescn Inc American) and test paper method measure basic blood glucose and glucose in urine respectively, then, press 150mg/kg body weight lumbar injection 3% alloxan.After 1 week when animal blood glucose concentration be greater than 11mol/L and glucose in urine ++ ++ (＞20g/L) time, diabetes model duplicates successfully.In the experimentation, diabetic animal is measured its glucose in urine variation every day, adopt the mode of replenishing injection alloxan or insulin (2u/ only) to keep its blood glucose at 11-16.5mmol/L.
Get imagineering's diabetes rat model, inject anesthesia with 3% pentobarbital sodium (25mg/kg) abdominal cavity after, hair is shaved at the back, routine disinfection.Each makes a call to a circular hole in spinal column both sides, Mus back of the body middle part with card punch under aseptic condition, be deep to subcutaneous, diameter 1.8cm, wound surface area 2.54cm
2The single cage in hemostasis back is fed, and quantitatively drinking-water and feeding obtain diabetes wound surface model.
2, experiment grouping and treatment
Get 40 of diabetes wound surface model mouses, be divided into five groups, I, II, III, IV and V group.Get 8 healthy non-diabetic rats, inject anesthesia with 3% pentobarbital sodium (25mg/kg) abdominal cavity after, hair is shaved at the back, routine disinfection.Each makes a call to a circular hole in spinal column both sides, Mus back of the body middle part with card punch under aseptic condition, be deep to subcutaneous, diameter 1.8cm, wound surface area 2.54cm
2The single cage in hemostasis back is fed, and quantitatively drinking-water and feeding obtain simple skin wound model, and not administration is as VI group (non-diabetic matched group).
I organizes (blank group): wound surface is coating not;
II organizes (low dosage experimental group): wound surface is coated with 10% calf-blood deprotein extract gel;
III organizes (middle dosage experiments group): wound surface is coated with 20% calf-blood deprotein extract gel;
IV organizes (high dose experimental group): wound surface is coated with 30% calf-blood deprotein extract gel;
V organizes (positive controls): wound surface coating Solcoseryl Jelly;
VI organizes (non-diabetic matched group): wound surface is coating not.
Every group of each 8 animal (16 wound surface).Clean wound surface every day, protect from infection, each group is all taked exposure method, will not wrap up.II-V organizes to have made in model and begins medication next day, evenly is coated with skim on wound surface, and thickness is about 1mm, changes dressings once successive administration 14 days in per 4 hours～6 hours.Write down the wound surface area of each experimental group and matched group in the 0th, 5,10,15 days respectively in administration, adopt computer aided video system to handle.Each is organized data and all represents with means standard deviation, with t check carrying out statistical analysis.
It is as shown in table 4 that each organizes the wound healing situation of rat:
Table 4 respectively organize the rat skin wound surface healing state (x ± SD, n=16)
| Group | 0d wound surface area (cm 2) | 5d wound surface area (cm 2) | 10d wound surface area (cm 2) | 15d wound surface area (cm 2) |
| I II III IV V VI | 2.54±0.00 2.54±0.00 2.54±0.00 2.54±0.00 2.54±0.00 2.54±0.00 | 2.22±0.31a 2.04±0.32 1.87±0.28 1.84±0.27 1.98±0.31 1.71±0.27 | 1.04±0.21b 0.73±0.22c 0.58±0.17d 0.53±0.15d 0.64±0.18d 0.43±0.17 | 0.45±0.13 0.29±0.11c 0.22±0.09d 0.20±0.07d 0.26±0.09c 0.00±0.00 |
Annotate: a and VI group be P＜0.05 relatively, and b and VI group be P＜0.01 relatively, and c and I group be P＜0.05 relatively, and d and I group be P＜0.01 relatively
Experimental result shows that the healing speed of diabetes rat (being the blank group) is obviously slow than non-diabetic rat, is 0.43cm at back 10 days average residual wound surface of non-diabetic rat of wound
2, the healing area reaches approximately 83%, and the average residual wound surface of diabetes rat is 1.04cm
2(P＜0.01), the healing area is 59% only, the wound surface of hindering back 15 days non-diabetic rats heals fully, and still there is about 0.45cm in diabetes rat
2Residual wound do not heal.Calf-blood deprotein extract gel can make the healing speed of diabetes rat accelerate, and after 2 weeks of treatment, low dose group residual wound area is 0.29cm
2, obviously dwindling (P＜0.05) than the blank group, middle dosage group and high dose group residual wound area are respectively 0.22cm
2And 0.20cm
2, compare difference with the blank group and have significance meaning (P＜0.01).Calf-blood deprotein extract gel group and Solcoseryl Jelly group be no significant difference relatively.The experimental group of various dose is compared, and middle and high dosage calf-blood deprotein extract gel group residual wound area is little than low dose group correspondingly in the identical time, but the difference not statistically significant.
Calf blood protein-removed extraction can not only improve picked-up and the utilization of histiocyte to oxygen and glucose, can also promote the migration and the propagation of fibroblast and vascular endothelial cell, microcirculation improvement, and protection ischemic tissue, thereby can promote wound healing.This experimental result shows that calf-blood deprotein extract gel can quicken the healing of rat diabetes skin ulcer wound surface, obviously dwindles the wound surface area.
The influence that embodiment 5, calf blood (protein removed) gel heal to the rabbit corneal epithelial defect
Experimental drug is with embodiment 3.
Adopt Japanese white big ear rabbit, body weight is 2.5～3.5Kg, male and female dual-purpose, healthy no oculopathy.Animal origin: Jinzhou Medical College's Experimental Animal Center (quality certification number: No. 038, distant real kinoplaszm card word).
1, animal model duplicates
Get healthy Japanese white big ear rabbit, the slow 5-8ml that annotates of 35% urethane ear vein, suction adds diethyl ether when insufficient.With the eye speculum fissura palpebrae of performing fighting, drip two of 2 1% tetracaine, making the topical anesthesia diameter is trepan tabulation lamination trace in the middle part of cornea of 8mm, use normal saline flushing behind 1% fluorescent staining, scrape whole epitheliums in the decyclization with lancet after appearing annellation, fluorescent staining is evenly painted discoid epithelial defect district of 8mm again, makes rabbit corneal epithelial defect model.
2, experiment grouping and Therapeutic Method
Get 40 of rabbit, as stated above the right eye of every rabbit is made the corneal epithelial defect model, be divided into 5 groups at random, 8 every group, eye drip treatment respectively.
I group: be the blank group, drip normal saline;
The II group: experimental group, use 20% calf-blood deprotein extract gel;
The III group: experimental group, use 50% calf-blood deprotein extract gel;
The IV group: experimental group, use 70% calf-blood deprotein extract gel;
The V group: positive matched group, use Solcoseryl Eye-Jel.
Each is organized medication and is each 1, and every day 4 times is to wound healing.Measured corneal epithelial wound healing area in per 24 hours as follows and calculate healing rate: after adopting every day fluorescence to dye the uniformly dyeing color,, average, press S=π R as the wound surface diameter with the length of 4 direction fluoresceins of pupillometry instrumentation amount colour attaching area
2, obtain the residue damaged area; Healing rate=healing area/initial damage area * 100%.Experiment gained data are represented with means standard deviation, with t check carrying out statistical analysis.The result is as shown in table 5:
Table 5 corneal epithelial wound healing area and healing rate be (mm relatively
2, X ± SD)
| Group | 24h | 48h | 72h | 96h | 120h |
| I group II group III group IV group V group | 17.22±2.06(34.26％) 24.31±1.53a(48.43％) 28.35±1.42abc(56.40％) 28.93±1.35abc(57.55％) 26.45±1.18ab(52.62％) | 26.76±1.53(53.23％) 33.85±1.33a(67.34％) 40.77±1.28abc(80.96％) 41.31±1.23abc(82.18％) 37.28±1.06ab(74.16％) | 36.23±1.64(72.07％) 44.78±0.74a(89.08％) 49.57±0.46abc(98.61％) 49.72±0.37abc(98.91％) 47.33±0.52ab(94.15％) | 46.18±0.84(91.86％) 50.27a(100％) 50.27a(100％) 50.27a(100％) 50.27a(100％) | 50.27(100％) - - - - |
A compares P＜0.01 with the I group; B compares P＜0.01 with the II group; C compares P＜0.01 with the V group
The wound healing area that shows basic, normal, high dosage group of calf-blood deprotein extract gel and Solcoseryl Eye-Jel group all healed in hindering the back all faster than the normal saline group on the 4th day fully, average specific blank group early 1 day, and difference has statistical significance.The healing speed of the middle and high dosage group of calf-blood deprotein extract gel is obviously faster than low dose group and Solcoseryl Eye-Jel group (P＜0.01, and there was no significant difference between the middle and high dosage experiments group.
The Main Ingredients and Appearance of calf-blood deprotein extract gel is a calf blood protein-removed extraction, and it is made up of small organic molecules such as inorganic ions and aminoacid, small-molecular peptides, nucleotide, oligosaccharide, lipids.These small-molecule substances that calf blood protein-removed extraction comprised can improve picked-up and the utilization of histiocyte to oxygen and glucose, can also promote the migration and the propagation of fibroblast and vascular endothelial cell, thereby can promote wound healing.This experimental result shows that calf-blood deprotein extract gel can obviously accelerate the healing of corneal epithelial wound.
Claims (7)
1, calf-blood deprotein extract gel is made by the raw material that comprises following compositions: calf blood (protein removed) extracting solution 20-50 weight portion, carbomer 0.5-1.2 weight portion, water for injection 45-80 weight portion; The quality percentage composition of solid content is 2%-7% in the described calf blood (protein removed) extracting solution.
2, calf-blood deprotein extract gel according to claim 1 is characterized in that: described raw material also comprises sodium hydroxide 0.2-0.5 weight portion.
3, calf-blood deprotein extract gel according to claim 2 is characterized in that: described raw material also comprises antibacterial 0.05-0.1 weight portion.
4, calf-blood deprotein extract gel according to claim 3 is characterized in that: described antibacterial is nipagin A, dimethyl fumarate or low-grade fatty acid ester.
5, calf-blood deprotein extract gel according to claim 4 is characterized in that: described nipagin A is an ethylparaben; Described low-grade fatty acid ester is a monoglyceride.
6, according to arbitrary described calf-blood deprotein extract gel among the claim 1-5, it is characterized in that: described calf blood (protein removed) extracting solution can be prepared as follows: adopt 1-6 monthly age children cattle venous blood, isolate serum, use the ethanol Deproteinization, ethanol is removed in decompression, adds the compound protein enzyme hydrolysis in remaining liquid, and supernatant is stayed in centrifugalize, described supernatant is held back with 5000 dalton's hollow fiber column ultrafilter, collects the filtrate that obtains and is the calf blood (protein removed) extracting solution; Wherein said during with the ethanol Deproteinization consumption of ethanol be that the quality percentage composition that doubles the serum volume is 96% ethanol; Wherein said decompression is removed ethanol for utilizing the rotary evaporation in vacuo instrument, and 37 ℃, 1 atmospheric pressure decompression removes ethanol; Wherein said adding compound protease is hydrolyzed to and adds compound protein enzyme hydrolysis 24 hours; Centrifugalize is 10000rpm, 4 ℃ after the enzyme hydrolysis of wherein said adding compound protein, centrifugal 30 minutes, stays supernatant.
7, according to arbitrary described calf-blood deprotein extract gel among the claim 1-5, it is characterized in that: described carbomer is 934GE, 910,934,934P, 940,941 or 1342.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010083636A1 (en) * | 2009-01-20 | 2010-07-29 | 沈阳兴齐制药有限公司 | An eyedrops of the deproteinized calf blood extract |
| CN102302514A (en) * | 2011-09-23 | 2012-01-04 | 沈阳斯佳科技发展有限公司 | Pig blood deproteinized extract gel and preparation method thereof |
| CN101518547B (en) * | 2008-02-27 | 2013-01-30 | 沈阳兴齐眼药股份有限公司 | Ophthalmic preparation combination containing deproteinised calf blood extract |
| CN107412116A (en) * | 2017-03-21 | 2017-12-01 | 陕西万维健康咨询服务有限公司 | A kind of reparation liquid of ox blood deproteinized, preparation method and applications |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR0163563B1 (en) * | 1994-03-23 | 1998-12-01 | 김종인 | Topical drug in combination for treatment of skin lesions |
| CN1062135C (en) * | 1996-11-27 | 2001-02-21 | 锦州医学院科技开发部 | Method for extracting deproteinized calf blood extract |
-
2005
- 2005-07-12 CN CNB2005100830412A patent/CN100360141C/en active Active
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101518547B (en) * | 2008-02-27 | 2013-01-30 | 沈阳兴齐眼药股份有限公司 | Ophthalmic preparation combination containing deproteinised calf blood extract |
| WO2010083636A1 (en) * | 2009-01-20 | 2010-07-29 | 沈阳兴齐制药有限公司 | An eyedrops of the deproteinized calf blood extract |
| CN101780105B (en) * | 2009-01-20 | 2011-09-28 | 沈阳兴齐制药有限公司 | Eye drop of deproteinized calf blood extractive |
| CN102302514A (en) * | 2011-09-23 | 2012-01-04 | 沈阳斯佳科技发展有限公司 | Pig blood deproteinized extract gel and preparation method thereof |
| CN102302514B (en) * | 2011-09-23 | 2013-05-01 | 沈阳斯佳科技发展有限公司 | Pig blood deproteinized extract gel and preparation method thereof |
| CN107412116A (en) * | 2017-03-21 | 2017-12-01 | 陕西万维健康咨询服务有限公司 | A kind of reparation liquid of ox blood deproteinized, preparation method and applications |
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