Summary of the invention
A kind of novel form that the purpose of this invention is to provide calf blood protein-removed extraction, calf-blood deprotein capsules.
Calf-blood deprotein capsules provided by the present invention is made up of capsule 's content and capsule shells; Described capsule 's content is made by following raw material: calf blood protein-removed extraction (solid content) 30-50 weight portion, starch 49-70 weight portion, antibacterial 0.1-1 weight portion; The quality percentage composition that described calf blood protein-removed extraction derives from solid content is the calf blood (protein removed) extracting solution of 2%-7%; Described capsule shells is an enteric capsule shell.
Calf-blood deprotein capsules can be hard capsule and also can be soft capsule.
The quality percentage composition that consists of of described calf blood protein extract is 60% K
+, Na
+And Ca
2+And the quality percentage composition is 40% nucleotide, aminoacid, small-molecular peptides and keto acid.Described calf blood protein-removed extraction 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 ".
Described calf blood protein-removed extraction is obtained at 40-50 ℃ of vacuum drying 20-24 hour by the calf blood (protein removed) extracting solution.
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; Consumption of ethanol is 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 starch is potato starch, corn starch, tapioca, wheaten starch etc.; Described corn starch is preferably pregelatinized Starch.
Described antibacterial can be nipagin A, dimethyl fumarate or low-grade fatty acid ester.Wherein, described nipagin A is an ethylparaben.
Another object of the present invention provides a kind of method for preparing above-mentioned calf-blood deprotein capsules.
The method for preparing calf-blood deprotein capsules provided by the present invention, be that quality percentage composition with solid content is the calf blood (protein removed) extracting solution of 2%--7%, 40-50 ℃ vacuum drying 20-24 hour, account for the ratio of 30-50 weight portion again according to calf blood protein-removed extraction (solid content), the starch that adds the 49-70 weight portion, 40-50 ℃ vacuum drying 20-24 hour, the antibacterial that adds the 0.1-1 weight portion again, cross 40 orders-80 mesh sieve, 40-50 ℃ of vacuum drying is 2 hours then, the enteric capsule shell of packing into obtains calf-blood deprotein capsules.
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, in remaining liquid, add the compound protein enzyme hydrolysis, supernatant is stayed in centrifugalize, and 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.
The consumption of said medicine is generally 900-1200mg calf blood protein-removed extraction every day/50Kg body weight, and be 10 to 20 days the course of treatment.
The present invention makes a kind of peroral dosage form of conveniently taking with calf blood protein-removed extraction: enteric coated capsule, both remedy liquid drugs injection and used inconvenient defective, overcome the unsettled deficiency of oral tablet again, can be used for treating 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).Zoopery and clinical trial prove that all it is evident in efficacy.
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 1,1000 calf-blood deprotein capsules of preparation
Capsule 's content is made up of following raw material: calf blood (protein removed) extracting solution (the quality percentage composition of solid content is 4%) 2500g, potato starch 149.75g, ethylparaben (ethyl hydroxybenzoate) 0.25g.
1, preparation calf blood (protein removed) extracting solution
2500g calf blood (protein removed) extracting solution (the quality percentage composition of solid content is 4%) is ZL96120777.9 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 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 are got 12L serum, add 24L 96% ethanol, constantly stir, left standstill 60 minutes, and made protein denaturation precipitation, then through 4 ℃ of centrifugal 30min of 20000rpm, abandon precipitation, 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, obtain 2500g calf blood (protein removed) extracting solution, the quality percentage composition of its solid content is 4%.
2, preparation calf-blood deprotein capsules content
The quality percentage composition of 2500g solid content is that 40 ℃ of vacuum dryings of calf blood (protein removed) extracting solution of 2%-7% are after 24 hours; to wherein adding 149.75 potato starch; continued 40 ℃ of vacuum dryings 24 hours; add the 0.25g ethyl hydroxybenzoate again, pulverize mixing; cross 80 mesh sieves; 40 ℃ of vacuum dryings 2 hours are granulated with granulator, obtain the calf-blood deprotein capsules content.
3, preparation calf-blood deprotein capsules
Preparation method according to conventional enteric coated capsule preparation, utilize JNP-1200 type capsule filling machine (purchase great magnificent pharmaceutical machine branch company) to fill the 250g capsule 's content in the long peak of space flight limited company Beijing, every contains the 250mg capsule 's content, obtains 1000 enteric coated capsulees.The calf-blood deprotein capsules of packing is poured in the Aluminium-coating Packer, with 10/plate of aluminum-plastic packaged one-tenth.
Embodiment 2,1000 calf-blood deprotein capsules of production
Capsule 's content is made up of following raw material: calf blood (protein removed) extracting solution (the quality percentage composition of solid content is 4%) 3125g, pregelatinized Starch 122.5g, dimethyl fumarate 2.5g.
1, preparation calf blood (protein removed) extracting solution
According to the patent No. is ZL 96120777.9, the method preparation of denomination of invention for describing in " a kind of method of extracting calf blood protein-removed extraction ":
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 are got 15L serum, add 30L 96% ethanol, constantly stir, left standstill 60 minutes, and made protein denaturation precipitation, then through 4 ℃ of centrifugal 30min of 20000rpm, abandon precipitation, obtain the 37.5L supernatant, utilize the rotary evaporation in vacuo instrument, 37 ℃, 1 atmospheric pressure decompression removes ethanol, in remaining liquid, add the 4.5g 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, obtain 3125g calf blood (protein removed) extracting solution, the quality percentage composition of its solid content is 4%.
2, preparation calf-blood deprotein capsules content
The quality percentage composition of 3125g solid content is that 40 ℃ of vacuum dryings of calf blood (protein removed) extracting solution of 4% are after 24 hours; to wherein adding the 122.5g pregelatinized Starch; continued 40 ℃ of vacuum dryings 24 hours; add the 2.5g dimethyl fumarate again, pulverize mixing; cross 80 mesh sieves; 40 ℃ of vacuum dryings 2 hours are granulated with granulator, obtain the calf-blood deprotein capsules content.
3, preparation calf-blood deprotein capsules
Preparation method according to conventional enteric coated capsule preparation, utilize JNP-1200 type capsule filling machine (purchase great magnificent pharmaceutical machine branch company) to fill the 250g capsule 's content in the long peak of space flight limited company Beijing, every contains the 250mg capsule 's content, obtains 1000 enteric coated capsulees.Pour the calf-blood deprotein capsules of packing into Aluminium-coating Packer, with 10/plate of aluminum-plastic packaged one-tenth.
Embodiment 3, zoopery
Experiment material
Calf blood (protein removed) injection (prescription: every contains solid content matter content is 4% calf blood (protein removed) extracting solution 5ml, and Jinzhou Aohong Pharmaceutical Co.,Ltd produces).
The preparation of calf-blood deprotein capsules: the calf-blood deprotein capsules content is with embodiment 1.
The preparation of starch enteric coated capsule: after the potato starch oven dry, use the enteric microcapsule packing with animal.
One, calf-blood deprotein capsules is to the influence of white mice normal pressure oxygen-resistant ability
Select 100 of healthy Kunming kind one-level white mice, body weight 18-22g (providing) by Jinzhou Medical College's Experimental Animal Center, male and female half and half, be divided into five groups, i.e. three experimental grouies of calf-blood deprotein capsules (I, II, III group), calf blood (protein removed) injection experimental group (IV group), starch capsule matched group (V group).I, II, III group are the high, medium and low dosage group of calf-blood deprotein capsules, give calf-blood deprotein capsules (in calf blood protein-removed extraction) 30mg/kg body weight, 15mg/kg body weight, 10mg/kg body weight every day respectively, once a day, administering mode is oral.The IV group is calf blood (protein removed) injection tail intravenously administrable, and dosage (by calf blood protein-removed extraction) and administration time are with dosage group in the calf-blood deprotein capsules, and administering mode is a tail vein injection, once a day.The V group gives 75mg starch/kg body weight every day for the blank group, and once a day, administering mode is oral.Each organized the white mice successive administration 11 days, after the last administration, all white mice were put in 30 minutes the 250ml wide mouthed bottle that 10g soda lime is housed, airtight normobaric hypoxia is observed the white mice time-to-live, measures the content of glucose and lactic acid in the cerebral tissue behind the mice dying.Normobaric hypoxia white mice time-to-live measurement result is as shown in table 1:
The table 1 normobaric hypoxia white mice time-to-live
Group | The example number | Route of administration | Dosage (calf blood protein-removed extraction meter) mg/kg body weight | Time-to-live (min) |
I II III IV V | 20 20 20 20 20 | Oral oral intravenous injection is oral | 30 15 10 15 75 (starch) | 46.63±7.03
a 46.37±8.20
a. 44.44±8.82
a 46.73±6.32
a.b. 35.65±5.61
|
Annotate: a, P<0.01 with the V group relatively; B, P>0.05 with the II group relatively
Table 1 shows that all calf-blood deprotein capsules groups and calf blood (protein removed) injection group white mice time-to-live all obviously be longer than the blank group, there was no significant difference between the high, medium and low dosage group (P>005), calf blood protein-removed extraction injection group and calf-blood deprotein capsules group there was no significant difference (P>005).
Normobaric hypoxia white mice brain lactic acid content measurement result is as shown in table 2:
Table 2 normobaric hypoxia white mice brain lactic acid content
Group |
The example number |
Route of administration |
Dosage (calf blood protein-removed extraction meter) mg/kgB.W |
Brain lactic acid content (mmol/g.wet.wt) |
I II III IV V |
20 20 20 20 20 |
Oral oral intravenous injection is oral |
30 15 10 15 75 (starch) |
9.68±0.61
a 10.01±0.56
a 11.3±0.80
a 9.84±0.41
a.b 12.36±0.85
|
Annotate: a, P<0.01 with the V group relatively; B, P>0.05 with the II group relatively
Table 2 shows that all calf-blood deprotein capsules groups and calf blood protein-removed extraction injection group white mice brain lactic acid content are starkly lower than matched group, there was no significant difference (P>005) between calf-blood deprotein capsules height, the middle dosage group, middle dosage group brain lactic acid content is starkly lower than low dose group, dosage group there was no significant difference (P>005) in calf blood protein-removed extraction injection group and the calf-blood deprotein capsules.
Normobaric hypoxia white mice brain glucose content measurement result is as shown in table 3
Table 3 normobaric hypoxia white mice brain glucose content (
Group | The example number | Route of administration | Dosage (calf blood protein-removed extraction meter) mg/kgB.W | Brain glucose content (mmol/g.wet.wt) |
I II III IV V | 20 20 20 20 20 | Oral oral intravenous injection is oral | 30 15 10 15 75 (starch) | 5.65±0.72
a 5.46±0.68
a. 4.65±0.60
a 5.66±0.45
a.b 3.74±0.18
|
Annotate: a, P<0.01 with the V group relatively; B, P>0.05 with the II group relatively
Table 3 shows, all calf-blood deprotein capsules groups and calf blood (protein removed) injection group white mice brain glucose content are apparently higher than matched group, there was no significant difference (p>005) between calf-blood deprotein capsules height, the middle dosage group, in dosage group brain glucose content apparently higher than low dose group, dosage group there was no significant difference (P>005) in calf blood (protein removed) injection group white mice brain glucose content and the calf-blood deprotein capsules.
Calf-blood deprotein capsules is a kind of oral formulations of calf blood protein-removed extraction, contains several physiological active substances, and this medicine can strengthen glucose and cell is gone in oxygen transport, increases the utilization of glucose.This experimental result shows that all normobaric hypoxia experimental group white mice brain lactic acid contents are starkly lower than matched group, prove that this capsule can promote normobaric hypoxia white mice brain cell to carry out aerobic metabolism, find simultaneously, dose-effect relationship is not obvious between calf-blood deprotein capsules height, the middle dosage group, in, obviously positive dose-effect relationship, calf blood (protein removed) injection and calf-blood deprotein capsules therapeutic equivalence arranged between the low dose group; Normobaric hypoxia white mice brain glucose content measurement result shows, all experimental group white mice brain glucose contents are all apparently higher than matched group, therefore, proof calf-blood deprotein capsules and calf blood (protein removed) injection can promote blood or other tissue glucose to transport to cerebral tissue, studies show that simultaneously, dose-effect relationship is not obvious between calf-blood deprotein capsules height, the middle dosage group, in, obviously positive dose-effect relationship, calf blood (protein removed) injection and calf-blood deprotein capsules therapeutic equivalence arranged between the low dose group; Normobaric hypoxia white mice time-to-live result of study shows, calf-blood deprotein capsules can prolong the normobaric hypoxia white mice time-to-live, dose-effect relationship is not obvious between the high, medium and low dosage group of calf-blood deprotein capsules, calf blood (protein removed) injection and calf-blood deprotein capsules therapeutic equivalence.