CN1314448C - Medical use and medicinal preparation of trichonema in preventing human and animals from diseases by immunologic inadequacy - Google Patents
Medical use and medicinal preparation of trichonema in preventing human and animals from diseases by immunologic inadequacy Download PDFInfo
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- CN1314448C CN1314448C CNB031110800A CN03111080A CN1314448C CN 1314448 C CN1314448 C CN 1314448C CN B031110800 A CNB031110800 A CN B031110800A CN 03111080 A CN03111080 A CN 03111080A CN 1314448 C CN1314448 C CN 1314448C
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- trichinella spiralis
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Abstract
The present invention discloses an application of trichina in the prevention and treatment of the diseases caused by the immunologic hypofunction of hunam beings and animals and corresponding medicinal preparations, namely trichina cell vaccines, trichina attenuated vaccines and trichina biologically active materials. The trichina cell vaccines, trichina attenuated vaccines and trichina biologically active materials can cause the inoculated human being or animals to generate complex natural antigen materials for enhancing the immunizing power of human beings and animals and further achieving the goals of preventing and treating the diseases caused by the immunologic hypofunction of hunam beings and animals by inoculating the trichina cell vaccines in an intramuscular inoculation mode, the trichina attenuated vaccines in a peroral inoculation mode, and the trichina biologically active materials in an infection inoculation mode or a peroral inoculation mode into human being or animals, and can simultaneously be used as vaccines for preventing and treating the trichinosis of human beings or animals.
Description
Technical field:
The present invention relates to the medical application that mao shape belongs to (Trichinella) nematicide, a kind of trichinellidae nematodes causes disease at control humans and animals immunologic hypofunction medical application is especially disclosed, corresponding pharmaceutical preparation is provided simultaneously, has belonged to biological pharmacy technical field.
Background technology:
For many years, people think that always parasite is the intravital insect of a kind of human or animal of colonizing in, but from dialectical materialist point of view, parasite also should have its useful available one side.In long-term biological evolution process, trichina(Trichinella spiralis) (Trichinella spiralis, Owen 1835) be called for short trichinella and in human or animal body, survive, many useful native antigen materials have been produced, though these complicated materials are not got its 26S Proteasome Structure and Function so far as yet clear, it has the effect of non-specific enhancing human or animal's immunologic function really.
Summary of the invention:
The invention discloses a kind of trichinellidae nematodes preventing and treating the medical application that the humans and animals immunologic hypofunction causes disease, corresponding pharmaceutical preparation is provided simultaneously.
Technical scheme of the present invention is as follows:
The preparation of trichina(Trichinella spiralis) cell vaccine
Aseptic collection trichina(Trichinella spiralis) 500-1, article 000, after, put into the RPMI1640 or the MEM culture medium (culture medium contains animal muscle protease digestion liquid) of improvement, after the polypide grinding, the centrifuging and taking supernatant carry out former be commissioned to train foster, treat that cell grows up to monolayer substantially after, outwell suspension and add the cultivation of going down to posterity of new culture medium, treat that cell reached for 30 generations, measure its rate of growth curve, cell colony formation rate and passage cell Chromosome number.After be used for the human or animal.
Trichina(Trichinella spiralis) causes weak Seedling preparation:
One of its preparation method
With trichina(Trichinella spiralis) by rat, rabbit or equus etc., continuous passage 10-15 generation, therebetween every 1 month with trichina(Trichinella spiralis) (500-10000 bar polypide) immunoprophylaxis, 3-5 time altogether.Collect its polypide and be used for the human or animal.
Two of preparation method
Directly collect the polypide that infects in the trichina(Trichinella spiralis) animal muscle, put into PBS, with the cobalt-60 (Co of 10-20 kiloroentgen (K γ) dosage
60) irradiation (or it is weak to adopt other physical method to cause), be used for the human or animal.
The preparation of trichina(Trichinella spiralis) bioactive substance:
Extract or carry out biochemical separation and Extraction adding the organic solvent of variable concentrations such as ethanol, petroleum ether and ethyl acetate etc. after the trichina(Trichinella spiralis) polypide fragmentation of the trichina(Trichinella spiralis) excrete thing of In vitro culture or collection, obtain different bioactive substances, through concentrate, packing after the sterilization treatment, be used for the human or animal.
Cause weak Seedling in human or animal body by intramuscular inoculation trichina(Trichinella spiralis) cell vaccine, per oral inoculation trichina(Trichinella spiralis), give the trichina(Trichinella spiralis) bioactive substance in human or animal body by injection or per os, make it produce complicated native antigen material, strengthen humans and animals immunity, and then reach the purpose that control humans and animals immunologic hypofunction or defective cause disease, also can prevent and treat the worm Seedling of humans and animals trichinosis simultaneously.
" indication ":
Be mainly used in immunologic hypofunction such as oncosis, early-stage cancer, sexual disorder and acquired immune deficiency syndrome (AIDS) or defective and cause prevention and treatment of diseases, also can prevent and treat the worm Seedling of humans and animals trichinosis simultaneously.
" using method ";
Trichina(Trichinella spiralis) cell vaccine: with 1 * 10
3-1 * 10
10Individual cell is given an intramuscular injection of animal and human, and 3 months repeat once at interval, and its dosage and number of times are on the state of an illness, age, sex, the body constitution of humans and animals and prevent and treat purpose etc. and decide.
Trichina(Trichinella spiralis) causes weak Seedling: with 50-10,000 polypide gives the human or animal once oral, and 3 months repeat once at interval, and its dosage and number of times are on the state of an illness, age, sex, the body constitution of humans and animals and prevent and treat purpose etc. and decide.
The trichina(Trichinella spiralis) bioactive substance:
According to the animal pattern experimental result, determine approach and consumption to people's medication.
Following pharmacological evaluation has confirmed that pharmaceutical preparation of the present invention causes the medical application of disease at control humans and animals immunologic hypofunction:
Case 1:
BALB/C Mus (male and female half and half, body gets 18-20g) with the health pure lines is divided into 7 groups at random, every group 10, except that the 1st group of injection HanksShi liquid 100 μ L, 2-4 organizes mice, give 50,100,200 respectively and cause weak trichina(Trichinella spiralis) polypide, after 3 weeks, contain 1 * 10 with 100 μ L respectively
6Individual SP2/0 myeloma cell is in the left hind subcutaneous injection of Mus.5-7 group mice contains 1 * 10 with 100 μ L respectively
6Individual SP2/0 myeloma cell is in the left hind subcutaneous injection of Mus, after 3 weeks respectively per os give 50,100,200 and cause weak trichina(Trichinella spiralis) polypide, result's application trichina(Trichinella spiralis) mice immunized in-vivo tumour volume before mouse-borne tumor and after the lotus tumor is significantly less than the control group mice gross tumor volume without the trichina(Trichinella spiralis) immunity, and indivedual mices do not see that tumor takes place, and illustrates that trichina(Trichinella spiralis) has very strong antitumor action (seeing Table 1).
Lotus tumor situation behind the trichina(Trichinella spiralis) polypide a little less than the inoculation of table 1. separate groups of mice causes
| Group | Tumor size (cm) | No tumor mouse number (only) |
| 1 2 3 4 5 6 7 | 2.00-2.50×3.00-3.30 1.20-1.50×1.10-1.20 1.10-1.30×1.10-1.20 1.10-1.30×1.00-1.15 1.00-1.20×1.00-1.10 0.98-1.10×0.90-1.10 0.90-1.10×1.00-1.15 | 0 1 1 2 2 2 2 |
Case 2:
Certain man, 71 years old, have a delicate constitution for a long time, calendar year 2001 December took trichina(Trichinella spiralis) on the 15th and cause weak 4000 polypides of Seedling, slight exothermic reaction was arranged after 20 days, adopt medicine to proofread and correct, transfer to normally, body constitution is recovered after three months, so far energetic, the hair blackening is not caught a cold.
Good effect of the present invention is: the trichina(Trichinella spiralis) cell vaccine: can reach the effect that trichina(Trichinella spiralis) causes weak Seedling immunity, and easily a large amount of preparations, cost is low, safety, curative effect height, sphere of action is wide.Trichina(Trichinella spiralis) causes weak Seedling: preparation is easy, and easy to use, cost is low, and is effective, and sphere of action is wide.The trichina(Trichinella spiralis) bioactive substance: composition is single, acts on clearly defined objective.
The specific embodiment:
Embodiment 1: the preparation of trichina(Trichinella spiralis) cell vaccine:
5000 of aseptic collection trichina(Trichinella spiralis)s, putting into mortar grinds polypide, make cell suspension with 0.25% trypsinization 30min, with PH7.2PBS centrifuge washing three times, the RPMI1640 culture fluid of reuse improvement is made suspension with the cell precipitation thing, and to be sub-packed in 3 capacity be 30ML and use in advance in the Tissue Culture Flask of glue primordial covering, beyond the Great Wall plug, cultivate in 37 ℃ of calorstats, when treating that cell grows up to monolayer substantially, outwelling suspension, (is to remove calcium salt in the HanksShi formula of liquid with GNH, magnesium salt and phosphate) washing is once, 0.25% trypsin solution immersion 20min with pre-temperature makes the cell desalination, centrifugal then, outwell supernatant, add the new culture fluid of 6ml, mixing, two bottles of packing continue to cultivate, and continuous passage cultivated for 30 generations, observe its rate of growth curve, the cell colony formation rate.
Embodiment 2: trichina(Trichinella spiralis) causes weak Seedling preparation
The rat of artificial challenge trichina(Trichinella spiralis) is put to death, get muscle and with meat mincer muscle is blended, put in the Digestive system (pepsin 0.5g, hydrochloric acid 0.7ml, water 100ml), polypide is collected in the digestion back, polypide is put into PBS carry out Co with 12 kiloroentgen dosage
60Preserve the irradiation back.
Embodiment 3: trichina(Trichinella spiralis) bioactive substance chemical extraction method
With 100, article 000, weigh after the polypide fragmentation, put into round-bottomed flask and add an amount of 70% soak with ethanol and spend the night post-heating backflow 2-3h, filter, concentrate ethanol extract, residual residue continues to use 70% alcohol heating reflux 4-5 time, merges the ethanol concentrated solution, residual residue reuse water boil 2 times, concentrate water cooking liquid, be incorporated in the ethanol concentrated solution, residue dry weigh standby.Get part ethanol concentrated solution and add an amount of petroleum ether extraction repeatedly, residual residue continues with ethyl acetate extraction repeatedly.It is standby to obtain total ethanol extract, total ethanol extract residue, ligroin extraction, acetic acid ether extract and 5 parts of residue at last.
Claims (3)
1, a kind of trichina(Trichinella spiralis) causes weak Seedling preparation method, may further comprise the steps: trichina(Trichinella spiralis) by rat, rabbit or equus, continuous passage 10-15 generation, was used the trichina(Trichinella spiralis) immunoprophylaxis therebetween every 1 month, 3-5 time altogether, collect its polypide and must cause weak Seedling.
2, a kind of trichina(Trichinella spiralis) causes weak Seedling preparation method, may further comprise the steps: directly collect the polypide that infects in the trichina(Trichinella spiralis) animal muscle, put into PBS, with the cobalt-60 of 10-20 kiloroentgen dosage shine cause a little less than, must cause weak Seedling.
3, the weak Seedling that causes of claim 1 or the preparation of 2 described methods is used in preparation treatment humans and animals immunologic hypofunction causes the medicine of disease.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031110800A CN1314448C (en) | 2003-02-25 | 2003-02-25 | Medical use and medicinal preparation of trichonema in preventing human and animals from diseases by immunologic inadequacy |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031110800A CN1314448C (en) | 2003-02-25 | 2003-02-25 | Medical use and medicinal preparation of trichonema in preventing human and animals from diseases by immunologic inadequacy |
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| CN1439428A CN1439428A (en) | 2003-09-03 |
| CN1314448C true CN1314448C (en) | 2007-05-09 |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114163525B (en) * | 2020-09-11 | 2023-11-10 | 吉林大学 | Egg yolk antibody for resisting excreted and secreted antigen of trichina, and preparation method and application thereof |
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2003
- 2003-02-25 CN CNB031110800A patent/CN1314448C/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 60Co γ射线辐照对旋毛虫影响的研究 刘景祥等，河南畜牧兽医，第15卷第1期 1994 * |
| 60Co γ射线辐照对旋毛虫影响的研究 刘景祥等，河南畜牧兽医，第15卷第1期 1994;旋毛虫幼虫分泌物抗原在小鼠体内诱生肿瘤细胞毒因子的研究 谢春玲等，中国人蓄共患病杂志，第10卷第6期 1994 * |
| 旋毛虫幼虫分泌物抗原在小鼠体内诱生肿瘤细胞毒因子的研究 谢春玲等，中国人蓄共患病杂志，第10卷第6期 1994 * |
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