CN101361881B - Preparation method as protein medicine lysozyme intestinal-tract sorbefacient - Google Patents
Preparation method as protein medicine lysozyme intestinal-tract sorbefacient Download PDFInfo
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- CN101361881B CN101361881B CN2008100426600A CN200810042660A CN101361881B CN 101361881 B CN101361881 B CN 101361881B CN 2008100426600 A CN2008100426600 A CN 2008100426600A CN 200810042660 A CN200810042660 A CN 200810042660A CN 101361881 B CN101361881 B CN 101361881B
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- lysozyme
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Abstract
The invention discloses the application of yam as an accelerant for intestinal absorption of protein drug lysozyme. According to a normal method, commercially available yam is decocted, soaked and boiled and then centrifugated at low speed to get supernatant liquid, namely, the part of aqueous phase extract of the yam is the yam accelerant which is used for lavage and medication of stomach together with lysozyme according to a certain proportion. The experiments show that the yam can be served as the accelerant of intestinal absorption of macromolecular protein drugs to greatly promote the intestinal absorption rate of lysozyme so as to increase the blood concentration of lysozyme. The results of the experiments also show that the yam accelerant generally has no influences on the blood-drug background of mouse lysozyme since the body condition of the mouse is normal, and can obviously and effectively improve the intestinal absorption of the protein drug lysozyme at the same time, thusbeing an effective intestinal absorption accelerant. The experiments also indicate that the yam accelerant has no obvious influences on the metabolism of endogenous protein lysozyme and exogenous protein lysozyme drugs.
Description
Technical field
The present invention relates to the Chinese medicine Rhizoma Dioscoreae as the new purposes, particularly Rhizoma Dioscoreae of intestinal absorption promoter preparation method as the intestinal absorption promoter of protein medicine lysozyme.
Background technology
Lysozyme lysozyme, molecular weight is about 14kDa, isoelectric point, IP 10.7, Britain bacteriologist Fleming at first found in nineteen twenty-two, was the alkaline enzyme of mucopolysaccharide in a kind of energy hydrolysis pathogenic bacterium.This enzyme extensively is present in the body fluid such as the Ovum Gallus domesticus album of birds and poultry, mammiferous tear, saliva, blood plasma, urine, milk, also contains this enzyme in the microorganism.Wherein abundant with Ovum Gallus domesticus album content, be a kind of alkaline protein, to thermally-stabilised.
The function of lysozyme: 1) as one of body nonspecific immunity factor, participate in the reaction of body panimmunity, replenished the intravital nonspecific immunity factor of animal, it can improve and strengthen the macrophage phagocytic digestive function, reduce the leukopenia that cytostatics causes, ward off in conjunction with antibacterial fat more, alleviate the endotoxin effect, thus enhancing human body immunity power.2) lysozyme has natural antibacterial and anti-inflammation functions, and it causes the gram-positive bacteria cell wall to break under the osmotic pressure effect by β-1,4 glycosidic bond in N-second phthalein 3-O-.alpha.-carboxyethyl-D-glucosamine. in the range of hydrolysed peptides polysaccharide and the N-second phthalein glucamine, and bacteriolysis takes place; In vivo under the environment; owing to have SigA sIgA, complement to participate in; lysozyme can also some gram negative bacteria of hydrolysis; as escherichia coli etc., thereby kill corrupt coccus in the intestinal, keep flora normalization in the intestinal; promote bacillus bifidus propagation; strengthen white clear sterilization albumen, the defense factor of Y-ball egg strengthens resistance infection.3) lysozyme also can directly combine with electronegative virus protein, forms double salt with DNA, RNA, apoprotein, makes virally inactivatedly, has certain anti-virus ability.And general antibacterial is felt simply helpless to virus.4) lysozyme also has the hematoblastic function of activation, can improve the tissue local disturbance of blood circulation, and the secretion pus strengthens local defense function, thereby embodies effects such as its hemostasis, detumescence.Can not cause the residual and drug accumulation of poisoning.
As national OTC chemical drugs, the oral buccal tablet of lysozyme and enteric coated capsule are widely-used, and compound recipe lysozyme eye drop, compound recipe lysozyme capsule and recombinant human lysozyme spray etc. also approved be the new drug of clinical research.But these oral class lysozyme medicines mostly are and directly play antibacterial and anti-inflammation functions in digestive tract, can't pass through gut barrier and enter blood, therefore can not effectively improve blood drug level, cause the interior absorption efficiency of body very low, bioavailability is very limited, can not give full play to the medical value of lysozyme.
The short-cut method that improves blood drug level is an intravenous drug, the excellent drug effect of existing multinomial patent of invention and reported in literature high level lysozyme blood drug level, as intravenous injection lysozyme 30000U/20g mice, the effective percentage of multiple inflammation such as treatment pneumonia, nephritis, cystitis, pharyngolaryngitis can reach 100%.But the injection system administration not only causes pain, particularly long-term repeat administration brings spirit and physiological immense pressure for patient, cause the local fat atrophy, cutaneous pigmentation etc., even may take place as serious drug anaphylaxiss such as Arthurs reactions, the immune complex that produces causes pathology damage in local vascular basement membrane deposition, and general immunity complex disease etc. (as serum sickness).
In recent years, the administering mode of non-injecting pathway has been subjected to paying close attention to widely, and wherein the oral administration mode is undoubtedly one of most convenient, safest approach.Yet oral the only way which must be passed---gastrointestinal tract is the maximum natural cover for defense of high molecular weight protein drug oral administration.The absorption experiment of isolated rat jejunum tissue shows that as a kind of protide macromolecular drug, the bioavailability of lysozyme only is 1.47 ± 0.20%, and absorption efficiency is then lower in the body.For effectively improving blood drug level, the exploitation oral Preparation can adopt various means to promote the intestinal absorption of protide macromolecular drug, and the promoter of wherein adding intestinal absorption in medicine is a kind of good strategy.
Summary of the invention
At the deficiency that prior art exists, the object of the present invention is to provide a kind of preparation method of intestinal absorption promoter of protein medicine lysozyme.
Goal of the invention of the present invention is achieved by the following technical solution:
From tcm theory, by screening tens of kinds of Chinese medicines, Chinese medicine to be studied as the intestinal absorption promoter of protein medicine lysozyme, experiment finds that Rhizoma Dioscoreae can obviously promote the intestinal absorption of lysozyme medicine, improves its bioavailability of medicament.
Acquired the Chinese medicine-Rhizoma Dioscoreae that can effectively promote the intestinal absorption of lysozyme by screening tens of kinds of Chinese herbal medicine, selecting." Chinese pharmacopoeia and " national Chinese herbal medicine compilation " record, Rhizoma Dioscoreae: sweet; Flat; Return spleen, lung, kidney channel, have the spleen reinforcing nourishing the stomach, the lung benefiting that promotes the production of body fluid, the effect of the kidney invigorating arresting seminal emission is a kind of common, also is one of Chinese medicine of using always.
The preparation method step of promoter of the present invention is as follows:
Get commercial goods Chinese crude drug Rhizoma Dioscoreae, fried according to a conventional method, soak about 0 minute in the ratio of every 100ml cold water soak 120g Rhizoma Dioscoreae then, fast fire boiled back slow fire fried about 20 minutes, get supernatant behind the low-speed centrifugal, promptly obtaining Rhizoma Dioscoreae water extract position is Rhizoma Dioscoreae promoter, and 4 ℃ of preservations are standby.
By lysozyme dosage is 0.5mg-5.0mg, and the Rhizoma Dioscoreae accelerator dosage is that the ratio use of 0.2ml-2.0ml is carried out gastric infusion to mice.Experimental results show that Rhizoma Dioscoreae can promote the intestinal absorption efficient of lysozyme significantly as the promoter of high molecular weight protein class medicine intestinal absorption really, improve the blood drug level of lysozyme.
Experimental result shows that also Rhizoma Dioscoreae promoter does not have influence substantially to mice lysozyme blood medicine background, and the mice body condition is normal, can promote to significant effective the intestinal absorption of protein medicine lysozyme simultaneously, is a kind of effective intestinal absorption promoter.
Experiment simultaneously shows that Rhizoma Dioscoreae promoter is to endogenous protein lysozyme and not obviously influence of extrinsic protein lysozyme drug metabolism.
Description of drawings
Fig. 1 is the facilitation that Rhizoma Dioscoreae absorbs lysozyme intestinal-tract.
The specific embodiment
How further specify the present invention below by specific embodiment realizes:
Embodiment 1
1, the preparation of lysozyme formulation and consumption
Hen's egg-white lysozyme (20000U/mg)
Preparation method: the oral pharmaceutical solutions of normal saline preparation 2mg/ml or 10mg/ml, 4 ℃ of preservations are standby.Using dosage: a 0.5mg-5.0mg/ mice
2, the preparation of Rhizoma Dioscoreae promoter and consumption
Commercial goods Chinese crude drug Rhizoma Dioscoreae, according to " usage and dosage of Chinese pharmacopoeia.
Preparation method: conventional method is fried, every 100ml cold water soak 120g Rhizoma Dioscoreae 30 minutes, and fast fire boiled back slow fire fried 20 minutes, got supernatant behind the low-speed centrifugal, promptly obtained Rhizoma Dioscoreae water extract position, and 4 ℃ of preservations are standby.
Using dosage: a 0.2ml-2.0ml/ mice
3, the mensuration of lysozyme blood drug level
Assay method: lysozyme micrococcus luteus colorimetry, the range of linearity are 2 mcg/ml～10 mcg/ml.Experimental result:
1, Rhizoma Dioscoreae promoter is improved the vivo effect of lysozyme blood drug level
Experimental design:
Experimental group---lysozyme+Rhizoma Dioscoreae promoter group
Matched group---1) Rhizoma Dioscoreae promoter group
2) lysozyme group
3) background group (mice endogenous lysozyme)
Each organizes sample size is 6 mices;
Experimental group oral administration gavage administration lysozyme 1mg/ and Rhizoma Dioscoreae promoter 1ml/; Matched group 1 oral administration gavage Rhizoma Dioscoreae promoter 1ml/ only; Matched group 2 oral administration gavage administration lysozyme 1mg/ only; Matched group 3 oral administration gavage normal saline 1ml/ only.
8 hours posterior orbits of administration are got blood, measure lysozyme blood drug level.
Shown in the following chart 1 of result:
Table 1:
The above results shows, there was no significant difference between each matched group, and experimental group lysozyme blood drug level reaches 15.68 mcg/ml, and blood drug level improves more than two times, compares significant difference with matched group.Only take matched group bioavailability＜1% of lysozyme, and experimental group bioavailability＞5%, experimental results show that Rhizoma Dioscoreae can promote the intestinal absorption efficient of lysozyme significantly as the promoter of high molecular weight protein class medicine intestinal absorption really, improve the blood drug level of lysozyme.
2, Rhizoma Dioscoreae promoter is to the influence of lysozyme endogenous levels
Because lysozyme also is one of mice body nonspecific immunity factor, is the endogenous material in the mouse blood, therefore need to investigate the influence situation of Rhizoma Dioscoreae promoter itself to the mice background level.
Mice gavages one week of Rhizoma Dioscoreae promoter continuously, and eye socket is got blood examination and surveyed lysozyme blood drug level.Find: the lysozyme blood drug level of Rhizoma Dioscoreae promoter group mice is 4.63 ± 0.8, mice background group is 3.89 ± 0.63 (see figure 1)s, there is not significant difference between the two, the result shows that Rhizoma Dioscoreae promoter does not have influence substantially to mice lysozyme blood medicine background, the mice body condition is normal, can promote to significant effective the intestinal absorption of protein medicine lysozyme simultaneously, be a kind of effective intestinal absorption promoter.
3, Rhizoma Dioscoreae promoter is to the metabolic influence of lysozyme blood medicine
In order to investigate Rhizoma Dioscoreae promoter,, detect lysozyme blood medicine and keep situation the mice track determining of experimental group oral administration after 8 hours and 16 hours to the metabolic influence of lysozyme blood medicine.Found that 8 hours blood drug level 15.68 ± 5.88 mcg/ml, blood drug level 5.69 ± 0.92 in 16 hours, mice background group blood drug level is respectively 3.96 ± 0.89 and 4.04 ± 0.42 simultaneously, the result shows, compare with 8 hours blood medicine maximum concentrations, the blood drug level of administration experimental mice after 16 hours obviously descends, the basic recovery normally, do not have significant difference with control group mice, illustrating can be by the mice homergy by the sorbefacient medicine lysozyme albumen of Rhizoma Dioscoreae.The situation of 10-12 hour human blood medicine of this situation and lysozyme buccal tablet effective drug duration is consistent.Therefore, Rhizoma Dioscoreae promoter is to endogenous protein lysozyme and not obviously influence of extrinsic protein lysozyme drug metabolism.
Claims (1)
1. the preparation method of protein medicine lysozyme intestinal-tract absorption enhancer, its step is as follows: commercially available Chinese medicine Rhizoma Dioscoreae is fried according to a conventional method, ratio in every 100ml cold water soak 120g Rhizoma Dioscoreae is soaked, it is fried that fast fire boils back slow fire, get supernatant behind the low-speed centrifugal, obtain Rhizoma Dioscoreae water extract position, promptly obtain the protein medicine lysozyme intestinal-tract absorption enhancer, this promoter is preserved down at 4 ℃.
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| CN2008100426600A CN101361881B (en) | 2008-09-09 | 2008-09-09 | Preparation method as protein medicine lysozyme intestinal-tract sorbefacient |
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| CN2008100426600A CN101361881B (en) | 2008-09-09 | 2008-09-09 | Preparation method as protein medicine lysozyme intestinal-tract sorbefacient |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101947315B (en) * | 2010-09-03 | 2013-01-30 | 上海沈李科工贸有限公司 | Application of Chinese medicinal bitter apricot seeds in lysozyme sodium alginate microspheres |
| CN102670677B (en) * | 2012-05-22 | 2014-04-02 | 上海大学 | New application of traditional Chinese medicine fennel |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1213282A (en) * | 1996-03-15 | 1999-04-07 | 尤尼金实验室股份有限公司 | Oral peptide pharmaceutical products |
| US20060068036A1 (en) * | 2002-12-31 | 2006-03-30 | National Yang-Ming University | Extract of Dioscorea sp. and the medical uses thereof |
| CN1805733A (en) * | 2003-04-15 | 2006-07-19 | 阿克塞斯有限公司 | Absorption enhancers such as e.g. BHT,BHA or propyl gallate |
-
2008
- 2008-09-09 CN CN2008100426600A patent/CN101361881B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1213282A (en) * | 1996-03-15 | 1999-04-07 | 尤尼金实验室股份有限公司 | Oral peptide pharmaceutical products |
| US20060068036A1 (en) * | 2002-12-31 | 2006-03-30 | National Yang-Ming University | Extract of Dioscorea sp. and the medical uses thereof |
| CN1805733A (en) * | 2003-04-15 | 2006-07-19 | 阿克塞斯有限公司 | Absorption enhancers such as e.g. BHT,BHA or propyl gallate |
Non-Patent Citations (2)
| Title |
|---|
| 徐延震.中药免疫机制初探.《中国兽药杂志》.1995,第29卷(第3期),第50-52页. * |
| 谭允育等.29种中药免疫药理活性的初步筛选.《北京中医药大学学报》.1994,第17卷(第2期),28-30. * |
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