CN102824417B - New method for treating helicobacter pylori related diseases - Google Patents

New method for treating helicobacter pylori related diseases Download PDF

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CN102824417B
CN102824417B CN 201210352856 CN201210352856A CN102824417B CN 102824417 B CN102824417 B CN 102824417B CN 201210352856 CN201210352856 CN 201210352856 CN 201210352856 A CN201210352856 A CN 201210352856A CN 102824417 B CN102824417 B CN 102824417B
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extract
herba polygoni
polygoni capitati
ethanol
helicobacter pylori
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CN102824417A (en
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王子厚
莫非
张淑华
张姝
胡伏莲
杨国珍
罗昭逊
夏曙华
孙朝琴
张然
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Zhejiang Public Health Pharmaceutical Co ltd
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王子厚
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Abstract

The invention relates to a new method for treating helicobacter pylori related diseases, in particular to application of polygonum capitatum or extract thereof in preparing a medicine for treating and/or preventing the diseases or symptoms related to helicobacter pylori infection. The invention discovers that the extract can effectively suppress the helicobacter pylori.

Description

The new method for the treatment of helicobacter pylori relevant disease
Technical field
The present invention relates to the field of Chinese medicines, particularly, the present invention relates to treat the new method of helicobacter pylori relevant disease, be specifically related to a kind of polygonaceae plant and extract thereof and be used for the treatment of the new method of helicobacter pylori relevant disease, particularly Chinese herbal medicine Herba Polygoni Capitati or its extract are used for the treatment of the new method of helicobacter pylori relevant disease.
Background technology
Herba Polygoni Capitati (Polygonum capitatum Buch.-Ham.ex D.Don) is a kind of Polygonaceae arsesmart, is perennial herb, another name Herba Polygoni Capitati, Herba Polygoni Capitati, Polygonum capitatum Buch, water silk ball, Flos Carthami Herba Violae, completely red etc.In areas such as Guizhou In China, Yunnan, Guangxi, Sichuan, Tibet, Hubei, Hunan and Jiangxi, distribution is all arranged, mainly be grown in hillside, mountain valley and be rich in the husky shale area in coal seam.Herb has the effect of heat-clearing and toxic substances removing, dissipating blood stasis, inducing diuresis for treating stranguria syndrome.According to 1963 " Guangxi Chinese medicinal herbal " second record, Herba Polygoni Capitati is mainly used in treating rheumatalgia; " Chinese medicine voluminous dictionary " first volume record in 1977, Herba Polygoni Capitati is mainly used in treating dysentery, nephritis, cystitis and lithangiuria etc.Among the people generally by Herba Polygoni Capitati with being decocted in water for oral dose, therapeutic effect is apparent in view, but takes inconvenience.Chinese patent application prospectus CN1054899A discloses a kind of miganling instant herbal medicine, syrup production technology in (Chinese Patent Application No. 90107810.7, open day on October 2nd, 1991).This production technology is to take four seasons grass as raw material water decoction 30-60 minute, extracting liquid filtering is concentrated, its supernatant concentrating under reduced pressure is become to cream, use again the 60-70% ethanol extraction, by the ethanol extract vacuum drying, obtain Herba Polygoni Capitati extractum, further be mixed with electuary or syrup, it is believed that this has removing toxic substances, dissipating blood stasis, diuresis, treating stranguria effect.
Recorded the Chinese patent medicine preparation of " the clear granule of pyretic stranguria " by name in Ministry of Health of the People's Republic of China's " drug standard-Traditional Chinese medicine historical preparation " (the 17) that Ministry of Health of the People's Republic of China's committee of pharmacopeia is compiled in 1998, it is made by Herba Polygoni Capitati being decocted with water to twice rear filtering and concentrating.
CN 1481832A (Chinese Patent Application No. 02129686.3, disclosed day on March 17th, 2004) and CN 1483466A (Chinese Patent Application No. 03146381.9, open day on March 24th, 2004) Herba Polygoni Capitati extract is disclosed, it is prepared by following steps basically: a. is by the fresh goods of Herba Polygoni Capitati herb or the dry product water decocts at twice or divide reflux, extract, two to three times with alcohol-water mixture, each 1-2 hour, merge decoction liquor, relative density during filtering and concentrating to 20 ° C is 1.2, and spray drying or drying under reduced pressure obtain; Perhaps, b. is obtained through carbon dioxide supercritical fluid extraction by Herba Polygoni Capitati herb and water extraction medicinal residues thereof.It is believed that this extract can be used for antibiotic, antiinflammatory, analgesia, diuresis, treatment urinary system calculus, treatment pyelonephritis and prostatitis.
Helicobacter pylori (Helicobacter pylori, in this article can referred to as Hp or HP) is a kind of microaerophilic gram negative bacteria be prevalent in people's gastric mucosa.Helicobacter pylori is the antibacterial of a kind of one pole, many flagellums, the blunt circle of end, helically bent, long 2.5~4.0 μ m, wide 0.5~1.0 μ m, often be typical helical form or arc on the gastric epithelial cell surface, while growing on solid medium, except typical form, sometimes shaft-like or spherical shape can appear.Helicobacter pylori surpasses 50% at the infection rate of global general population, and the population that does not almost have in the world a kind of chronic infectious disease can make the whole world surpass half is infected, and developing country is higher than developed country.China belongs to developing country, and the Helicobacter pylori infection rate is high.Digest by Chinese Medical Association the Helicobacter pylori infection Epidemiological study demonstration that relates to the general population at 20 ,40Ge of provinces and cities centers, the whole nation that disease branch helicobacter pylori group is done: China's Helicobacter pylori infection rate is 40-90%, average out to 59%; Existing disease infection rate is 42-64%, average 55%.It is believed that Hp infects and the main pathogenic of multiple prevalent upper gastrointestinal tract disease as peptic ulcer, gastritis, gastric mucosa dependency lymphoma (MALT) and gastric cancer etc.Known for example following disease or disease infect relevant with Hp directly or indirectly: gastric mucosa dependency lymphoma, peptic ulcer, gastric ulcer, duodenal ulcer, acute or chronic gastritis, superficial gastritis, atrophic gastritis, gastric cancer, cardiovascular disease, coronary heart disease, arteriosclerosis, cerebral infarction, hypertension, anemia, iron deficiency anemia, recurrent peptic ulcer, Mucosal atrophy, intestinal epithelial metaplasia, and thrombocytopenic purpura (Baidupedia: http://baike.baidu.com/view/36784.htm, 2011-09-15), can also comprise the complication of following these diseases or disease, suppress the relapse rate that helicobacter pylori is conducive to reduce peptic ulcer, blocking-up or delay further developing of Mucosal atrophy and intestinal epithelial metaplasia.Within 1994, World Health Organization (WHO)/international cancer research institution (WHO/IARC) is decided to be I class carcinogen by helicobacter pylori.Hp anti-infective therapy take antibiotic to be three or the quadruple chemotherapy on basis, but combines and be widely used in anti-Hp treatment along with Multiple Classes of Antibiotics, and Hp is day by day serious to antibiotic drug resistance problem, has become the main cause of eradication therapy failure.
The new treatment of helicobacter pylori relevant disease is expected to be useful in this area.
Summary of the invention
The object of the invention is to for a kind of disease relevant with Helicobacter pylori infection or method of disease for the treatment of and/or preventing is provided clinically.The inventor is surprisingly found out that, Herba Polygoni Capitati or its extract are suppressing helicobacter pylori and treating and/or preventing the disease relevant with Helicobacter pylori infection or disease has the effect of desirable.The present invention is based on this and be accomplished.
For this reason, first aspect present invention provides Herba Polygoni Capitati or the purposes of its extract in the medicine for the preparation for the treatment of and/or preventing the disease relevant with Helicobacter pylori infection or disease.
According to the purposes of first aspect present invention, wherein said Herba Polygoni Capitati is fresh goods or the dry product of Herba Polygoni Capitati.
According to the purposes of first aspect present invention, wherein said Herba Polygoni Capitati is herb, aerial parts, leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
According to the purposes of first aspect present invention, wherein said Herba Polygoni Capitati is the aerial parts that is selected from Herba Polygoni Capitati.
According to the purposes of first aspect present invention, wherein said Herba Polygoni Capitati is leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
According to the purposes of first aspect present invention, wherein said extract is that Herba Polygoni Capitati water, ethanol, ethanol water or its combination are as solvent, through extracting the extract obtained; Or Herba Polygoni Capitati is through the extract of carbon dioxide supercritical fluid extraction method acquisition.
According to the purposes of first aspect present invention, wherein said extract is the extract of the ethanol water of Herba Polygoni Capitati.In one embodiment, described ethanol water is 30 ~ 99% ethanol waters.In one embodiment, described ethanol water is 50 ~ 98% ethanol waters.In one embodiment, described ethanol water is 60 ~ 98% ethanol waters.In one embodiment, described ethanol water is 70 ~ 95% ethanol waters.
According to the purposes of first aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: decocted and/or reflux, extract, concentrated extracting solution, drying by medicinal material of polygonum capilalum water, ethanol or ethanol water.
According to the purposes of first aspect present invention, wherein said extract obtains with the carbon dioxide supercritical fluid extraction Herba Polygoni Capitati.In one embodiment, wherein said extract is the extract prepared according to the method comprised the steps: use the carbon dioxide supercritical fluid extraction medicinal material of polygonum capilalum, and/or with the carbon dioxide supercritical fluid extraction medicinal material of polygonum capilalum through water, ethanol or ethanol water decocts and/or reflux, extract, obtains medicinal residues.
Purposes according to first aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: get medicinal material of polygonum capilalum, add 5 ~ 20 times (for example 5 ~ 15 times, for example 8 ~ 12 times) water, ethanol or ethanol water decoct and/or reflux, extract, 1 ~ 5 time (for example 2 ~ 3 times), each 0.5 ~ 5 hour (for example 0.5 ~ 3 hour, for example 1 ~ 3 hour), collecting decoction, filter, filtrate decompression is concentrated into the thick paste that relative density is 1.1 ~ 1.5 (25 ° of C) (for example 1.2 ~ 1.4), drying, obtain.
Purposes according to first aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: get medicinal material of polygonum capilalum, add 5 ~ 20 times (for example 5 ~ 15 times, for example 8 ~ 12 times) water, decoct and extract 0.5 ~ 5 hour (for example 0.5 ~ 3 hour, for example 1 ~ 3 hour) 80 ~ 100 ° of C left and right; Filter, filtrate decompression is condensed into paste; Doubly measure 50 ~ 90% alcohol reflux 2-3 time with 2-5, by ethanol extract in 70-90 ° of C except ethanol, vacuum drying, obtain.
Purposes according to first aspect present invention, the wherein said disease relevant with Helicobacter pylori infection or disease include but not limited to, gastric mucosa dependency lymphoma, peptic ulcer, gastric ulcer, duodenal ulcer, acute or chronic gastritis, superficial gastritis, atrophic gastritis, gastric cancer, cardiovascular disease, coronary heart disease, arteriosclerosis, cerebral infarction, hypertension, anemia, iron deficiency anemia, recurrent peptic ulcer, Mucosal atrophy, intestinal epithelial metaplasia and thrombocytopenic purpura, and their complication.
The known disease relevant with Helicobacter pylori infection or disease can adopt the drug combination therapeutic scheme, for example three or the tetrad therapeutic scheme, for example, the combined treatment of amoxicillin+clarithromycin+metronidazole, perhaps proton pump inhibitor (omeprazole, lansoprazole and/or pantoprazole)+combined treatment of amoxicillin+clarithromycin, the perhaps combined treatment of omeprazole+clarithromycin+amoxicillin+ranitidine, perhaps on their basis, be aided with gastric mucosa protectant such as but not limited to sucralfate, the combined treatment of dioctahedral smectite etc.And the present invention finds that Herba Polygoni Capitati extract has good bacteriostatic activity, particularly surprisingly find that Herba Polygoni Capitati extract is also effective to the pathogen of some anti-amoxicillin, clarithromycin and/or metronidazole, therefore can expect, Herba Polygoni Capitati extract of the present invention can be used with these antiviral drug regimens or substitute them and use.Therefore, according to the purposes of first aspect present invention, also comprise one or more other medicine in wherein said medicine, this other medicine is selected from: antimicrobial drug, proton pump inhibitor, gastric mucosa protectant etc.Described antimicrobial drug such as but not limited to: penicillins is amoxicillin for example, and Macrolide is clarithromycin for example, and the nitre imidazoles is metronidazole for example.Described proton pump inhibitor is such as but not limited to omeprazole, lansoprazole, pantoprazole.Described gastric mucosa protectant is such as but not limited to sucralfate, dioctahedral smectite etc.How those skilled in the art are combined Herba Polygoni Capitati of the present invention or its extract and described other medicine according to existing Knowledge.
Second aspect present invention provides and has been used for the treatment of and/or disease that prevention is relevant with Helicobacter pylori infection or the pharmaceutical composition of disease, wherein comprises Herba Polygoni Capitati or its extract, and optional pharmaceutically acceptable carrier or excipient.
According to the pharmaceutical composition of second aspect present invention, wherein said Herba Polygoni Capitati is fresh goods or the dry product of Herba Polygoni Capitati.
According to the pharmaceutical composition of second aspect present invention, wherein said Herba Polygoni Capitati is herb, aerial parts, leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
According to the pharmaceutical composition of second aspect present invention, wherein said Herba Polygoni Capitati is the aerial parts that is selected from Herba Polygoni Capitati.
According to the pharmaceutical composition of second aspect present invention, wherein said Herba Polygoni Capitati is leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
According to the pharmaceutical composition of second aspect present invention, wherein said extract is that Herba Polygoni Capitati water, ethanol, ethanol water or its combination are as solvent, through extracting the extract obtained; Or Herba Polygoni Capitati is through the extract of carbon dioxide supercritical fluid extraction method acquisition.
According to the pharmaceutical composition of second aspect present invention, wherein said extract is the extract of the ethanol water of Herba Polygoni Capitati.In one embodiment, described ethanol water is 30 ~ 99% ethanol waters.In one embodiment, described ethanol water is 50 ~ 98% ethanol waters.In one embodiment, described ethanol water is 60 ~ 98% ethanol waters.In one embodiment, described ethanol water is 70 ~ 95% ethanol waters.
According to the pharmaceutical composition of second aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: decocted and/or reflux, extract, concentrated extracting solution, drying by medicinal material of polygonum capilalum water, ethanol or ethanol water.
According to the pharmaceutical composition of second aspect present invention, wherein said extract obtains with the carbon dioxide supercritical fluid extraction Herba Polygoni Capitati.In one embodiment, wherein said extract is the extract prepared according to the method comprised the steps: use the carbon dioxide supercritical fluid extraction medicinal material of polygonum capilalum, and/or with the carbon dioxide supercritical fluid extraction medicinal material of polygonum capilalum through water, ethanol or ethanol water decocts and/or reflux, extract, obtains medicinal residues.
Pharmaceutical composition according to second aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: get medicinal material of polygonum capilalum, add 5 ~ 20 times (for example 5 ~ 15 times, for example 8 ~ 12 times) water, ethanol or ethanol water decoct and/or reflux, extract, 1 ~ 5 time (for example 2 ~ 3 times), each 0.5 ~ 5 hour (for example 0.5 ~ 3 hour, for example 1 ~ 3 hour), collecting decoction, filter, filtrate decompression is concentrated into the thick paste that relative density is 1.1 ~ 1.5 (25 ° of C) (for example 1.2 ~ 1.4), drying, obtain.
Pharmaceutical composition according to second aspect present invention, wherein said extract is the extract prepared according to the method comprised the steps: get medicinal material of polygonum capilalum, add 5 ~ 20 times (for example 5 ~ 15 times, for example 8 ~ 12 times) water, decoct and extract 0.5 ~ 5 hour (for example 0.5 ~ 3 hour, for example 1 ~ 3 hour) 80 ~ 100 ° of C left and right; Filter, filtrate decompression is condensed into paste; Doubly measure 50 ~ 90% alcohol reflux 2-3 time with 2-5, by ethanol extract in 70-90 ° of C except ethanol, vacuum drying, obtain.
Pharmaceutical composition according to second aspect present invention, the wherein said disease relevant with Helicobacter pylori infection or disease include but not limited to, gastric mucosa dependency lymphoma, peptic ulcer, gastric ulcer, duodenal ulcer, acute or chronic gastritis, superficial gastritis, atrophic gastritis, gastric cancer, cardiovascular disease, coronary heart disease, arteriosclerosis, cerebral infarction, hypertension, anemia, iron deficiency anemia, recurrent peptic ulcer, Mucosal atrophy, intestinal epithelial metaplasia and thrombocytopenic purpura, and their complication.
According to the pharmaceutical composition of second aspect present invention, wherein also comprise one or more and be selected from following medicine: antimicrobial drug, proton pump inhibitor, gastric mucosa protectant etc.Described antimicrobial drug such as but not limited to: penicillins is amoxicillin for example, and Macrolide is clarithromycin for example, and the nitre imidazoles is metronidazole for example.Described proton pump inhibitor is such as but not limited to omeprazole, lansoprazole, pantoprazole.Described gastric mucosa protectant is such as but not limited to sucralfate, dioctahedral smectite etc.
According to the pharmaceutical composition of second aspect present invention, it is the dosage form of oral or drug administration by injection.In one embodiment, described pharmaceutical composition is the form of tablet, capsule, granule, pill, oral solutions, injection (liquid drugs injection and/or powder pin) etc.
Third aspect present invention provides the disease relevant with Helicobacter pylori infection or the method for disease for the treatment of and/or preventing, and the method comprises to the Herba Polygoni Capitati of the administration effective dose that needs are arranged or its extract.
Method according to third aspect present invention, the wherein said disease relevant with Helicobacter pylori infection or disease include but not limited to, gastric mucosa dependency lymphoma, peptic ulcer, gastric ulcer, duodenal ulcer, acute or chronic gastritis, superficial gastritis, atrophic gastritis, gastric cancer, cardiovascular disease, coronary heart disease, arteriosclerosis, cerebral infarction, hypertension, anemia, iron deficiency anemia, recurrent peptic ulcer, Mucosal atrophy, intestinal epithelial metaplasia and thrombocytopenic purpura, and their complication.
According to the method for third aspect present invention, wherein also before using described Herba Polygoni Capitati or its extract, simultaneously or use afterwards one or more and be selected from following medicine: antimicrobial drug, proton pump inhibitor, gastric mucosa protectant etc.Described antimicrobial drug such as but not limited to: penicillins is amoxicillin for example, and Macrolide is clarithromycin for example, and the nitre imidazoles is metronidazole for example.Described proton pump inhibitor is such as but not limited to omeprazole, lansoprazole, pantoprazole.Described gastric mucosa protectant is such as but not limited to sucralfate, dioctahedral smectite etc.
Method according to third aspect present invention, wherein said mammal is that any position at health is (particularly in digestive tract, more particularly in the harmonization of the stomach duodenum) infect helicobacter pylori or have by the tendency of Helicobacter pylori infection or possible any mammal, comprise the mankind, and performing animal (such as but not limited to: pig, cattle, sheep, horse etc.), experimental animal (such as but not limited to: Canis familiaris L., cat, rat, mice, rabbit, monkey etc.), companion animals (such as but not limited to: pet dog, pet cat etc.), wild animal (such as but not limited to: tiger, ape, monkey, panda etc.), animal for display (mammal that for example raise in zoo).
Arbitrary embodiment of applicable equally other the arbitrary embodiment of arbitrary technical characterictic that arbitrary embodiment of either side of the present invention or this either side has or other either side, as long as they can be not conflicting, certainly, at where applicable each other, necessary words can be done suitably to modify to individual features.Below with characteristics, be further described to various aspects of the present invention.
All documents that the present invention quotes from, their full content is incorporated to this paper by reference, and if, when the expressed implication of these documents and the present invention are inconsistent, be as the criterion with statement of the present invention.In addition, various terms and phrase that the present invention uses have the general sense of well known to a person skilled in the art, nonetheless, the present invention still wishes at this, these terms and phrase to be described in more detail and to explain, the term of mentioning and phrase, if any inconsistent with known implication, are as the criterion with the implication that the present invention was explained.
In the present invention, unless the linguistic context of addressing clearly contains other implication, otherwise the Herba Polygoni Capitati of mentioning refers to Polygonaceae arsesmart Herba Polygoni Capitati (Polygonum capitatum Buch.-Ham.ex D.D on), comprise its fresh goods or dry product, comprise its herb, aerial parts, leaf, flower, stem, seed or its combination.
In the present invention, phrase " disease relevant with Helicobacter pylori infection or disease " can mean any disease, and its origin cause of formation is directly or indirectly relevant with Helicobacter pylori infection.In addition, especially, " disease relevant with Helicobacter pylori infection or disease " in most cases directly translates into gastropathy, such as gastritis, gastric ulcer etc., therefore, the present invention uses the disease of Chinese medicine involved in the present invention or its extract for treating or prevention, comprises directly or indirectly disease or the disease relevant with Helicobacter pylori infection of any origin cause of formation; And; as long as the tested individual body endogenous cause of ill origin cause of formation directly or indirectly and Helicobacter pylori infection and the body illness or the disease that cause; use Chinese medicine involved in the present invention or its extract for treating or prevention can demonstrate clinical effectiveness, all fall into protection scope of the present invention.For example certain style is as the patient; that health occurs is any (certain/some) the stomach discomfort symptom; and infect helicobacter pylori is arranged in its stomach after testing; use Chinese medicine involved in the present invention or its extract for treating or prevention can demonstrate clinical effectiveness; and this treatment or prevent related therapeutic use and pharmaceutical composition and Therapeutic Method, all fall into protection scope of the present invention.
In the present invention, term " extraction " can be the dipping under room temperature, or extraction at elevated temperatures (for example decoct and/or reflux), or the combination of these modes of operation.Can further include extract is further processed, for example further carry out purification, such as desolventizing, to remove impurity by means of precipitation matter, solvent extraction, resin absorption separation etc.
As described herein, term " extract " will comprise the extract of any purity that can be used in the present invention purpose, and according to the present invention, spirit is appreciated that the DNA purity of extract of the present invention can change in the larger context to those skilled in the art.For example Herba Polygoni Capitati of the present invention extracts the extract obtained, difference according to different process conditions, the 1kg Herba Polygoni Capitati for example, through extracting the arbitrary amount between 10g to 500g that can obtain (10g to 200g, 10g to 150g for example, 10g to 100g for example) that is the extract of different purity, those skilled in the art can, according to different needs, allocate the compositions that is applicable to needs with the extract of different purity.In some embodiments of the present invention, described extract can be called extract powder.
In the present invention, can use the present composition of effective dose to be applied to the tested individuality that needs are arranged.As described herein, term " effective dose " refers to the purpose dosage that can realize preventing and/or treating situation of the present invention, obstacle, disease or disease in the experimenter.Those skilled in the art, according to the context of the invention, can easily determine the using dosage of the present composition.Especially, this is according to the present invention, term " effective dose " can be understood as the present composition with reasonable effect/risk of being applicable to any therapeutic treatment and/or prevention than the q.s that treats and/or prevents described situation, obstacle, disease or disease.But the total consumption per day that it should be understood that the present composition can maked decision in the medical judgment scope reliably by those skilled in the art.For any concrete experimenter, the concrete horizontal fibrous root of prevention effective dose is determined according to many factors, and described factor comprises experimenter's age, body weight, general health situation, sex and diet; The concrete compositions adopted; Other therapeutic active substance that is used in combination with the present composition or uses simultaneously; And the known similar factor of medical field.
The invention provides and comprise the compositions formulated together with one or more nontoxic physiology's acceptable carriers.Described compositions can be mixed with solid or liquid form especially specially for Orally administered or for rectal administration, or is mixed with for injection and uses.
Include but not limited to capsule, tablet, pill, powder and granule for Orally administered solid dosage forms.In this type of solid dosage forms, reactive compound can be accepted excipient or carrier with the physiology of at least one inertia and mix as sodium citrate or dicalcium phosphate and/or following material: a) filler or extender are as starch, lactose, sucrose, glucose, mannitol and silicic acid; B) binding agent is as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and Radix Acaciae senegalis; C) wetting agent is as glycerol; D) disintegrating agent is as agar, calcium carbonate, Rhizoma Solani tuber osi or tapioca, alginic acid, some silicate and sodium carbonate; E) the solution blocker is as paraffin; F) absorb accelerator as quaternary ammonium compound; G) wetting agent is as spermol and glyceryl monostearate; H) adsorbent is as Kaolin and bentonite and i) lubricant is as Pulvis Talci, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulphate and their mixture.In the situation that capsule, tablet and pill also can comprise buffer agent in described dosage form.
Comprise the acceptable Emulsion of physiology, solution, suspensoid, syrup and elixir for Orally administered liquid dosage form.Liquid dosage form also can contain this area inert diluent commonly used except containing active component, for example water or other solvents, solubilizing agent and emulsifying agent be ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3 butylene glycol, dimethyl formamide, oils (particularly Oleum Gossypii semen, Oleum Arachidis hypogaeae semen, Semen Maydis oil, germ oil, olive oil, Oleum Ricini and Oleum sesami), glycerol, oxolane alcohol, Polyethylene Glycol and the fatty acid ester of sorbitan and their mixture for example.
Since the people such as nineteen eighty-three Marshall isolate Hp, domestic and international many scholars have carried out a large amount of research to Hp, find to eradicate the infection of Hp and can treat chronic gastritis, reduce peptic ulcer relapse rate, block or delay further developing of Mucosal atrophy and intestinal epithelial metaplasia.But kill the basic principle that Hp becomes the medical treatment Hp of Gastroenterology dept. related diseases.Developed for many years the multiple therapeutic scheme for Hp, standard scheme is that PPI adds two kinds of antibiotic triple therapies at present.But when infecting with antibiotic therapy Hp, in body, normal flora also is killed or suppresses, and causes the dysbacteriosis at gastrointestinal tract, pars oralis pharyngis, vagina and each position of skin, causes superinfection.Conventional triple therapy descends gradually to its eradication rate.Therefore find the alternative triple therapy of suitable medicine and seem particularly important.
Herba Polygoni Capitati is the conventional Chinese medicine of minority area, within 2000, enters National essential drugs list.It is one of kind in " the seven large Chinese medicine industrial chains " of Guizhou Province's modernization of Chinese medicine " the six large Miao Ethnomedicine " given priority to before 2010 and Guizhou Tenth Five-Year Plan Period emphasis cultivation and development.From the Herba Polygoni Capitati herb, isolated organic compound is individual more than totally six ten at present, mainly contains flavonoid, aldehyde, ketone, alcohols and terpenoid, and main effective ingredient is gallic acid and flavonoid.Be mainly used at present the symptoms such as heat-clearing and toxic substances removing, inducing diuresis for treating stranguria syndrome, pyelonephritis, lithangiuria, cystitis, dysentery, rheumatalgia, traumatic injury, urinary tract infection, skin infection eczema.
Although people know, the water extract of Herba Polygoni Capitati all has certain antibacterial activity in vivo and in vitro, especially gonococcus is had to stronger antibacterial activity, (the minimal inhibitory concentration scope is 8-32g/L, and meansigma methods is 11.2g/L).Secondly staphylococcus aureus, staphylococcus epidermidis, escherichia coli, dysentery bacterium, Bacillus proteus and Klebsiella Pneumoniae are also had to stronger antibacterial activity.Bacillus fragilis in part fungus and anaerobe, dyspepsiacoccus, peptostreptococcus and propionibacterium acnes, propionibacterium granulosum etc. are all had to certain antibacterial action.Yet the inventor finds unexpectedly, Herba Polygoni Capitati is effective for this intractable pathogenic microorganism of helicobacter pylori and caused associated conditions thereof.
The present invention adopts agar dilution to carry out antibacterial activity test.Reliable and stable for experimental result, strict experiment contrast, repetition and parallel laboratory test are set up in this experiment, and each sample all carries out three times identical experiment continuously, and each experiment is all at least inoculated two flat boards to each concentration.Result shows, three times result is consistent, and negative control (not adding bacterium) is without growth, growth control (i.e. not dosing) well-grown, solvent control (only solubilizer does not add sample)) have no the growth inhibited phenomenon.We are by the In Vitro Bacteriostasis experiment discovery of Herba Polygoni Capitati extract, and it is to two strain helicobacter pylori ATCC700392 and ATCC700824 fungistatic effect obvious (MIC is 4mg/mL).Herba Polygoni Capitati extract is to clinical isolates strain Hp B-1 type (to the amoxicillin drug resistance), Hp B-2 type (to the clarithromycin drug resistance), Hp B-3 type (to the metronidazole drug resistance), Hp B-4 type is (to amoxicillin, clarithromycin, the bacterial strain of three kinds of equal drug resistances of medicine of metronidazole), Hp B-5 type is (to amoxicillin, clarithromycin, the equal responsive bacterial strain of three kinds of medicines of metronidazole) in the antibacterial activity test of each 3 strain clinical strains, for experimental result reliable and stable, strict experiment contrast is set up in this experiment, repeat and parallel laboratory test, each sample all carries out three times identical experiment continuously, each experiment is all at least inoculated two flat boards to each concentration.Result shows, three times result is consistent, and it is consistent with the pre-stage test result that negative control (not adding bacterium) has no growth inhibited phenomenon, bacterial strain contrast (do Hp international standard check order strain ATCC700392) without growth, growth control (i.e. not dosing) well-grown, solvent control (only solubilizer) simultaneously.
Result of the present invention shows, Herba Polygoni Capitati extract all has stronger vitro inhibition Hp effect to helicobacter pylori international standard order-checking strain ATCC700392 and ATCC700824 two strain bacterium.In addition, there is stronger vitro inhibition Hp effect in the experiment of Herba Polygoni Capitati extract to clinical isolates strain Hp B-1 type (to the amoxicillin drug resistance), Hp B-2 type (to the clarithromycin drug resistance), Hp B-3 type (to the metronidazole drug resistance), HpB-4 type (to the bacterial strain of amoxicillin, clarithromycin, three kinds of equal drug resistances of medicine of metronidazole), Hp B-5 type (to amoxicillin, clarithromycin, the equal responsive bacterial strain of three kinds of medicines of metronidazole).
The present invention, by the experiment of Herba Polygoni Capitati extract antibacterial activity in vitro, is surprisingly found out that it has the activity of anti-helicobacter pylori.For example in one embodiment, by Herba Polygoni Capitati extract respectively to two strain helicobacter pylori (Helicobacter pylori, be called for short H.pylori, or Hp) ATCC700392 and ATCC700824 carry out the In Vitro Bacteriostasis experiment, at negative control (not adding bacterium), without growth, growth control (i.e. not dosing) well-grown, solvent control (only solubilizer), have no under the condition of growth inhibited phenomenon, the high dilution of the sample of the integral asepsis of take growth is the minimum inhibitory concentration (minimal inhibitory concentration, MIC) of this bacterial strain to respective sample.In another embodiment, Herba Polygoni Capitati extract is carried out to the In Vitro Bacteriostasis experiment to these five types each 3 strain clinical separation strains respectively, in the growth of negative control (not adding bacterium) nothing, growth control (i.e. not dosing) well-grown, solvent control (only solubilizer) has no the growth inhibited phenomenon, sample contrast (simultaneously making first stage H-2-65 sample) is consistent with the first stage experimental result, bacterial strain contrast (simultaneously being Hp international standard order-checking strain ATCC700392) and pre-stage test are as a result under uniform condition, the high dilution of the sample of the integral asepsis of take growth is the minimum inhibitory concentration of this bacterial strain to respective sample.The inventor finds: fungistatic effect obvious (MIC is≤4mg/mL) when (1) Herba Polygoni Capitati extract acts on Hp ATCC700392 and ATCC700824.(2) fungistatic effect obvious (MIC be 4 μ g/mLs) of Herba Polygoni Capitati extract to the clinical separation 3 strain bacterium of Hp B-1 type (to the amoxicillin drug resistance); Fungistatic effect obvious (MIC is 4 μ g/mL) to the clinical separation 3 strain bacterium of Hp B-2 type (to the clarithromycin drug resistance); The fungistatic effect of the clinical separation 3 strain bacterium of Hp B-3 type (to the metronidazole drug resistance) is (MIC is≤2 μ g/mL) obviously; The fungistatic effect of the clinical separation 3 strain bacterium of Hp B-4 type (to the bacterial strain of amoxicillin, clarithromycin, three kinds of equal drug resistances of medicine of metronidazole) is (MIC is≤8 μ g/mL) obviously, and the fungistatic effect of the clinical separation 3 strain bacterium of Hp B-5 type (to amoxicillin, clarithromycin, the equal responsive bacterial strain of three kinds of medicines of metronidazole) is (MIC is≤4 μ g/mL) obviously; (3) DH-2-65, the DH-2-24 fungistatic effect obvious (MIC is≤4 μ g/mL) to the Hp clinical separation strain.Thus, Herba Polygoni Capitati extract all has stronger In Vitro Bacteriostasis effect to two strain helicobacter pylori international standard order-checking strain ATCC700392 and ATCC700824 two strain bacterium; There is stronger vitro inhibition Hp effect in the experiment of Herba Polygoni Capitati extract to the clinical isolates strain, thereby be expected to become a kind of brand-new antibacterials particularly can be used for the treatment of and/or prevention is relevant with Helicobacter pylori infection disease or disease.
The accompanying drawing explanation
Fig. 1 has shown the result of Herba Polygoni Capitati extract to the helicobacter pylori In Vitro Bacteriostasis.
Fig. 2 has shown that Herba Polygoni Capitati extract processes the Electronic Speculum figure of antibacterial microscopic morphology after helicobacter pylori.
The specific embodiment
Further illustrate the present invention below by specific embodiment/experimental example, still, should be understood to, these embodiment and experimental example are only used for the use specifically described more in detail, and should not be construed as for limiting in any form the present invention.
The present invention carries out generality and/or concrete description to the material and the test method that use in test.Although, for to realize that many materials and operational approach that the object of the invention is used are well known in the art, the present invention still does to describe in detail as far as possible at this.It will be apparent to those skilled in the art that hereinafter, if not specified, material therefor of the present invention and operational approach are well known in the art.
a, Preparation Example part
embodiment 1: prepare Herba Polygoni Capitati aerial parts water extract
Get Herba Polygoni Capitati aerial parts (dry product) 220kg, clean, add 7 times of water gagings, heated and boiled 1.5 hours, filter, and adds 6 times of water gagings in medicinal residues, boils 1.0 hours, filters.Merging filtrate, being concentrated into relative density is that 1.2 (20 ℃) obtain extractum, spray drying obtains extract powder 23.1kg (yield 10.4%), is the Herba Polygoni Capitati extract of the present embodiment: brown ceramic powder; Every g sample is equivalent to crude drug in whole 8.7g; The employing spectrophotometer method is measured, flavones content: 18.21mg/g, gallic acid content: 17.82mg/g.
Get 1 part of above extract powder, add 2 parts of starch, 2 parts of microcrystalline Cellulose, mix homogeneously, encapsulated, be pharmaceutical composition of the present invention.
embodiment 2: prepare Herba Polygoni Capitati herb water extract (spray drying)
The method of reference example 1, difference is to replace the Herba Polygoni Capitati aerial parts to be extracted with Herba Polygoni Capitati herb (fresh goods), obtains the Herba Polygoni Capitati extract of the present embodiment.Yield 8.12%.
embodiment 3: prepare Herba Polygoni Capitati root 95% ethanol extraction
Get Herba Polygoni Capitati dry root 10kg, clean, put in the rustless steel concentration tank, add 95% ethanol of 7.5 times of amounts, reflux, extract, 1.5 hours, filter.Medicinal residues add 7.5 times of amount 95% ethanol, and reflux, extract, 1.0 hours, filter.Merge filtrate twice, concentrate to obtain extractum, drying under reduced pressure, obtain the present embodiment extract, 0.34kg.
embodiment 4: prepare Herba Polygoni Capitati stem 75% ethanol extraction
The method that reference example 3 is identical, difference is extracted the dry stem of Herba Polygoni Capitati with 75% ethanol, obtain the present embodiment extract.Yield 5.74%.
embodiment 5: prepare headdress flower Folium Polygonum 60% ethanol extraction
The method that reference example 3 is identical, difference is extracted the Herba Polygoni Capitati dried leaves with 60% ethanol, obtain the present embodiment extract.Yield 8.62%.
embodiment 6: prepare Herba Polygoni Capitati seed 30% ethanol extraction
The method that reference example 3 is identical, difference is extracted the Herba Polygoni Capitati dry seed with 30% ethanol, obtain the present embodiment extract.Yield 13.4%.
embodiment 7: prepare Herba Polygoni Capitati soaking with sugar thing
Get Herba Polygoni Capitati aerial parts (dry product), be ground into fine powder, mix (1:1, weight) with saturated sucrose solution and mix (attention prevents growth of microorganism), place more than 3 months.The mixture of placing 3 months makes solid-liquid separation through extruding, and makes the solid content water content lower than 15%, and the gained leaching liquid is used for biological test example part of the present invention as extract of the present invention.Hereinafter in test example, result shows the effectiveness of this leaching liquid, shows purposes that medicinal material of polygonum capilalum can directly (for example directly be mixed and made into honeyed pill with sugar) and expects for the present invention.
embodiment 8: prepare Herba Polygoni Capitati herb 7% ethanol extraction
Get Herba Polygoni Capitati herb 10kg, clean, put in the rustless steel concentration tank, add 7-8 doubly to measure 80% ethanol, reflux, extract, 1.5 hours, filter.Medicinal residues add 5 times of amount 60% ethanol, and reflux, extract, 1.5 hours, filter.Merge filtrate twice, concentrate to obtain extractum, drying under reduced pressure obtains extract 0.74kg.
embodiment 9: prepare Herba Polygoni Capitati herb water decoction-alcohol sedimentation solution powder
Get Herba Polygoni Capitati herb 10kg, clean, add 350kg water, heated and boiled 1.5 hours, shift supernatant, adds 150kg water in residue, boils 1.0 hours.Merge decoction liquor, filtering and concentrating to relative density is that 1.04 (24 ℃) obtain extractum 16kg.Adding 95% ethanol to be adjusted to containing the alcohol amount is 60%, quiet to 24 hours, divides and gets solution.To precipitation, adding 95% ethanol to be adjusted to containing the alcohol amount is 60%, quiet to 24 hours, divides and gets solution, merges solution twice, flings to ethanol, and 80 ℃ of drying under reduced pressure obtain Herba Polygoni Capitati herb water decoction-alcohol sedimentation solution powder 0.78kg, as the extract of the present embodiment.
embodiment 10: prepare Herba Polygoni Capitati herb water decoction-alcohol sedimentation precipitated powder
Get Herba Polygoni Capitati herb 10kg, clean, add 350kg water, heated and boiled 1.5 hours, shift supernatant, adds 150kg water in residue, boils 1.0 hours.Merge decoction liquor, filtering and concentrating to relative density is that 1.04 (24 ℃) obtain extractum 16kg.Adding 95% ethanol to be adjusted to containing the alcohol amount is 60%, quiet to 24 hours, divides and gets solution.To precipitation, adding 95% ethanol to be adjusted to containing the alcohol amount is 60%, quiet to 24 hours, divides and gets solution, merges solution twice, flings to ethanol, and 80 ℃ of drying under reduced pressure obtain Herba Polygoni Capitati herb water decoction-alcohol sedimentation solution powder 0.78kg; Be deposited in 80 ℃ of drying under reduced pressure by 16kg extractum with the post-processing step gained, obtain Herba Polygoni Capitati herb water decoction-alcohol sedimentation precipitated powder 0.49kg, as the extract of the present embodiment.
embodiment 11: prepare the former medicated powder carbon dioxide supercritical fluid extraction of Herba Polygoni Capitati extract
Get the former medicated powder 10kg of Herba Polygoni Capitati aerial parts, clean, drop in supercritical extraction reactor, to extraction kettle and two extraction-container heating, to basin, carry out cooling, when temperature reaches predetermined temperature, open dioxide bottle and supply gas, when pressure reaches predetermined pressure, start cycling extraction, adjust flux, and add entrainer, constant temperature and pressure extraction 2 hours.The liquid pressure-reducing obtained is concentrated and obtains extractum 0.17kg, as the extract of the present embodiment.
embodiment 12: prepare Herba Polygoni Capitati medicinal residues carbon dioxide supercritical fluid extraction extract
Get Herba Polygoni Capitati medicinal residues (the embodiment total medicinal residues of 1 gained and the total medicinal residues of embodiment 8 gained are with the mixture of weight ratio 1:1) 10kg, drop in supercritical extraction reactor, to extraction kettle and the heating of two extraction-containers, basin is carried out cooling, when temperature reaches predetermined temperature, opening dioxide bottle supplies gas, when pressure reaches predetermined pressure, start cycling extraction, adjust flux, and adding entrainer, constant temperature and pressure extracts 2 hours.By the concentrated extractum 0.131kg that obtains of the liquid pressure-reducing obtained.
embodiment 13: prepare Herba Polygoni Capitati medicinal residues carbon dioxide supercritical fluid extraction extract
Get Herba Polygoni Capitati aerial parts (dry product) 30kg, the water that adds 8 times of amounts, between 80-100 ° of C, decoct 60 minutes, extracting solution enters precipitating jar, again to the water that adds 4 times of amounts of medical material in decoction pot, between 80-100 ° of C, decoct 60 minutes, extracting solution enters precipitating jar and merges with extracting solution last time, get supernatant and send into vacuum concentration pan after clarifying, be evaporated to paste under 60-80 ° of C, then this paste is sent in the ethanol extraction pot, measure 65% ethanol extractions 2 times with 3 times, again ethanol extract is sent in vacuum concentration pan, remove ethanol under 70-90 ° of C, concentrating under reduced pressure, dry, obtain the present embodiment extract, yield 3.82%.
embodiment 14: prepare Herba Polygoni Capitati extract
Herba Polygoni Capitati aerial parts (dry product) 10kg, add 80 liters, water, reflux, extract, 2 times, and each 2h, merge extractive liquid,, standing over night, centrifugal.Macroporous adsorbent resin AB-8 (10kg) on filtrate, first wash 2 column volumes with water, discards eluent, then wash 3 column volumes with 85% ethanol, collects eluent, and constant pressure and dry, obtain Herba Polygoni Capitati extract.
After testing, this extract is 15% containing total flavones, and gallic acid is 2.5%.
embodiment 15: prepare Herba Polygoni Capitati extract
Herba Polygoni Capitati aerial parts (dry product) 10kg, add 10 times of amount 50% ethanol, reflux, extract, 3 times, each 1.5h, merge extractive liquid,, decompression recycling ethanol to ethanol content lower than 10%, upper macroporous adsorbent resin NKA-9, first wash 2 column volumes with water, discards eluent, use again 2 column volumes of 10% ethanol elution, discard eluent, then wash 3 column volumes with 80% ethanol, collect eluent, drying under reduced pressure, obtain Herba Polygoni Capitati extract.
After testing, this extract is 41% containing total flavones, and gallic acid is 11%.
embodiment 16: prepare Herba Polygoni Capitati extract
Herba Polygoni Capitati aerial parts (dry product) 10kg, add 10 times of amount 95% ethanol, reflux, extract, 2 times, each 2h, merge extractive liquid,, decompression recycling ethanol to ethanol content lower than 10%, upper macroporous adsorbent resin HPD400, first wash 2 column volumes with water, discard eluent, then wash 2 column volumes with 90% ethanol, collect eluent, spray drying, obtain Herba Polygoni Capitati extract.
After testing, this extract is 39% containing total flavones, and gallic acid is 7%.
embodiment 17: prepare Herba Polygoni Capitati extract
Herba Polygoni Capitati aerial parts (dry product) 10kg, add 10 times of amount 95% ethanol, reflux, extract, 2 times, each 2h, merge extractive liquid,, decompression recycling ethanol to ethanol content lower than 10%, upper macroporous adsorbent resin HPD100, first wash 2 column volumes with water, discard eluent, then wash 2 column volumes with 90% ethanol, collect eluent, drying under reduced pressure, obtain Herba Polygoni Capitati extract.
After testing, this extract is 32% containing total flavones, and gallic acid is 8%.
embodiment 18: prepare Herba Polygoni Capitati extract
Herba Polygoni Capitati aerial parts (dry product) 10kg, add 8 times of water gagings, reflux, extract, 2 times, and each 2h, merge extractive liquid,, standing over night, centrifugal.Macroporous adsorbent resin ADS-17 on filtrate, first wash 2 column volumes with water, discards eluent, then use 2 column volumes of 10% ethanol elution, discards eluent, then wash 2 column volumes with 90% ethanol, collects eluent, and drying under reduced pressure, obtain Herba Polygoni Capitati extract.
After testing, this extract is 37% containing total flavones, and gallic acid is 9%.
b, biological test example part
test example 1: Herba Polygoni Capitati extract (embodiment 1) was to helicobacter pylori type strain and clinical minute antibacterial activity from bacterial strain detects
1) experimental strain
reference culture:two strain bacterial strains used are helicobacter pylori ATCC700392 and helicobacter pylori ATCC700824 international standard order-checking strain, and purchased from ATCC (American Type Culture Collecti), passage number is within 10 generations.
the clinical isolates strain:for the strain of helicobacter pylori clinical isolates, all that the Guiyang Medical College Digestive System Department will be done gastroscopy and turn out to be peptic ulcer and chronic gastritis, under mirror, biopsy mucosa RUT is determined as positive patient simultaneously, and the gastric antrum section mucosa of its biopsy is separated to the Hp bacterial strain.The evaluation of Hp bacterial strain adopts the authentication method of internationally recognized helicobacter pylori, is divided into morphology, urease, oxidase, hydrogen peroxide test, serological test.Clinically be separated to five kinds of bacterial strains (name) and characteristic is respectively:
Hp B-1 type: it is to the amoxicillin drug resistance,
Hp B-2 type: it is to the clarithromycin drug resistance,
Hp B-3 type: it is to the metronidazole drug resistance,
Hp B-4 type: its bacterial strain to amoxicillin, clarithromycin, three kinds of equal drug resistances of medicine of metronidazole,
Hp B-5 type: it is to amoxicillin, clarithromycin, the equal responsive bacterial strain of three kinds of medicines of metronidazole.
2) some main agents that test is used
Colombia's blood agar and brain heart infusion powder derive from Britain OXOID, aseptic defiber Sanguis caprae seu ovis derives from Beijing swallow and gives birth to the rich bio tech ltd of political affairs, oxidase reagent, urease reagent, catalase reagent and E-test strip derive from Biomerieux SA, and Lyphocin (Fujisawa), amphotericin, sulfanilamide (TMP) and polymyxin B derive from U.S. Sigma company.
3) test specimen
The present invention's part embodiment gained Herba Polygoni Capitati extract.
4) preparation of main agents and material:
4.1) mix antibiotic: the method for by specification is prepared
Lyphocin (Fujisawa) 30mg, TMP25mg, polymyxin B20mg, both sexes rhzomorph 20mg, sterile purified water 100ml.
4.2) preparation of common Colombia blood plate:
The preparation of by specification method, take Colombia's agar basis 8g in triangular flask, be placed in the 250ml conical flask, add the mono-steaming of 180ml water, after 121 ℃ of 20min high pressure steam sterilizations, be cooled to 56 ℃ of left and right, add in superclean bench and mix antibiotic 2ml and aseptic defiber Sanguis caprae seu ovis 20ml, adjusting pH value is 7.2, and vibration mixes rear quick pour plate, the about 20ml of every ware, put into 4 ℃ of Refrigerator stores after the agar cooled and solidified, in 3 days, use, the random ware of selecting is put into 37 ℃ of incubators cultivation 24h simultaneously, guarantees that this batch of culture dish of asepsis growth can be used.
4.3) containing the preparation of sample blood plate:
The sample blood plate preparation of variable concentrations: take Colombia blood agar powder 3.9g in conical flask, add the mono-steaming of 89ml water, after high pressure steam sterilization, be cooled to 50 ℃ of left and right, the blood plate of the different gradient concentrations of each sample adds the respective sample mother solution from high to low successively according to concentration, making its final concentration is 8mg/mL, 4mg/mL, 1024 μ g/mL, 512 μ g mL, 256 μ g/mL, 128 μ g/mL, 64 μ g/mL, 32 μ g/mL, add the aseptic defiber Sanguis caprae seu ovis of 1ml mixing antibiotic and 10ml simultaneously, vibration mixes rear quick pour plate (plate diameter is 90mm), after the agar cooled and solidified, can use.
4.4) containing the preparation (solvent control) of solvent blood plate: method, with the preparation containing the sample blood plate, adds the dimethyl sulfoxine of respective volume or dehydrated alcohol to substitute the sample mother solution, and the distilled water of usining does not need preparation as solvent.
5) experimental technique
5.1) the recovery cultivation of helicobacter pylori type strain and the evaluation of bacterial strain
Take out helicobacter pylori ATCC700392 and helicobacter pylori ATCC700824 reference culture from-80 ℃ of refrigerators, being placed in 37 ℃ of water baths dissolves, getting bacterium liquid with the inoculating loop libation at an ancient wedding ceremony evenly coats containing mixing on antibiotic Colombia blood plate, be placed in anaerobism and cultivate box, and place two bags of micro-aerobic aerogenesis bag (85%N in box 2, 10%CO 2, 5%O 2), the rearmounted 37 ℃ of cultivation 72-96h of sealing.This two strains reference culture is carried out to cultural character, Gram's staining, urease evaluation, catalase evaluation, oxidase evaluation.
5.2) preparation of bacterium liquid: the bacterium colony on blood plate or lawn are scraped to pre-cooling containing in 2ml brain heart infusion meat soup pipe, after centrifuge washing three times, be resuspended in 4 ℃ of brain heart infusion meat soups, after this bacterium liquid is done to 10 times, 100 times, 1000 times dilutions, survey the OD600 value of bacterium liquid on ultraviolet spectrophotometer, OD value and the bacterial concentration of result between 0.1-1.0 has linear relationship preferably, and the computing formula of bacterium liquid hold-up is as follows: bacterium amount/ml=2.2 * 10 8* OD 600value * extension rate.The dilution of bacterium liquid is obtained to 10 8cFU/mL.Completed the preparation of bacterium liquid and inoculation in 15 minutes, in seeded process, bacterium liquid need remain on 4 ℃.
5.3) experiment of the antibacterial activity in vitro of Herba Polygoni Capitati extract
5.3.1) coating of bacterium liquid: drawing 200 μ l concentration is 10 8cFU/mL bacterium liquid, evenly be coated with at agar surface with the L-type rod.All be coated with 2 flat boards by each concentration of every kind of sample.Establish negative control (not adding bacterium), growth control (not adding laboratory sample), each two flat boards of solvent control simultaneously.
5.3.2) cultivate: method to specifications, be placed in anaerobism and cultivate box, and place two bags of micro-aerobic aerogenesis bags in box, seal latter 37 ℃ and cultivate observed result after 72-96h.The antibacterial activity in vitro experiment of each sample repeats twice.
5.3.3) the Herba Polygoni Capitati extract fungistatic effect judges: under negative control (not adding bacterium) asepsis growth, growth control (not adding laboratory sample) well-grown, the condition of solvent control without the growth inhibited phenomenon, the high dilution of the sample of the integral asepsis of take growth is the MIC of this bacterial strain to respective sample.
6) result of experiment:
6.1) evaluation of helicobacter pylori
Two strain helicobacter pylori ATCC700392 and ATCC700824 and clinical separation strain are placed under micro-aerobic condition on common Colombia blood plate cultivates 72-96 hour, and bacterium colony is the tip-like water white transparency, smooth moistening, neat in edge, projection, be little water droplet sample, the lawn canescence, hair oil light.
Biochemical test: the urease test positive, the oxidase test positive, the catalase test positive; In microscope oil Microscopic observation, typical sea-gull shape, S shape, Gram's staining feminine gender.
6.2) antibacterial activity in vitro of Herba Polygoni Capitati extract test experimental result is in Table 1.
The extract of table 1, the embodiment of the present invention 1 is to helicobacter pylori In Vitro Bacteriostasis result (MIC)
Figure BDA00002167364700191
Figure BDA00002167364700201
In the parallel test of the present embodiment, simultaneous verification amoxicillin, clarithromycin, the metronidazole three sensitivity to seven kinds of helicobacter pylori in upper table, in result and common the detection, gained is basically identical, three kinds of medicines are all responsive to ATCC700392 and ATCC700824, and Hp B-1 is to the amoxicillin drug resistance, to other two kinds of medicaments insensitives; Hp B-2 is to the clarithromycin drug resistance, to other two kinds of medicaments insensitives; Hp B-3 is to the metronidazole drug resistance, to other two kinds of medicaments insensitives; Hp B-4 is to amoxicillin, clarithromycin, three kinds of equal drug resistances of medicine of metronidazole; Hp B-5 is all responsive to amoxicillin, clarithromycin, three kinds of medicines of metronidazole.
In addition, each test group antibacterial (ATCC700392) growing state of perusal, the results are shown in Figure 1.Wherein Fig. 1 a is the growth control group that antibacterial grows in culture medium, shows that bacterial growth is normal, is homodisperse small particles; Fig. 1 b is antibacterial having added the solvent matched group of growing for the culture medium of the solvent of preparating liquid, shows that bacterial growth is normal, is homodisperse small particles; Fig. 1 c is the plate that does not add antibacterial in parallel test, and negative matched group shows without bacterial growth, without the small particles of typical bacteria growth; Fig. 1 d, 1e, 1f are respectively that the embodiment of the present invention 1 extract adds to latter 2 days, 3 days, 4 days macroscopic results in the plate of bacterial growth, be presented at 2 days antibacterials obviously few than the growth control group, within the 3rd day, only appearance is individual else is dispersed in the antibacterial small particles, and can not see antibacterial fully by the 4th day, similar with the plate of negative control group.
In addition, in above test, in ATCC700392 bacterium group, get blank 0 day, blank 4 days, Herba Polygoni Capitati is processed 2 days, Herba Polygoni Capitati and is processed the culture of 4 days, carries out respectively electron microscopic examination, and result is shown in respectively Fig. 2 a, 2b, 2c and 2d.As seen from the figure, that the antibacterial of 0 day culture of blank is is shaft-like, bagel sample, the variform thalline smooth surface such as U-shaped, neat in edge, and (Fig. 2 is a) for the electron density homogeneous; The antibacterial of 4 days cultures of blank is rod-short, bagel sample, the variform such as U-shaped, thalline smooth surface, neat in edge (Fig. 2 b); The antibacterial that Herba Polygoni Capitati is processed 2 days cultures is the variforms such as rod-short, U-shaped, bagel sample, has part thalline surface that the performances (Fig. 2 c) such as depression, disappearance are arranged; It is serious that the antibacterial that Herba Polygoni Capitati is processed 4 days cultures is the form part thalline surface depressions such as elongated rod shape, screw shaped, an end curve inwardly, and division (Fig. 2 d) appears in part thalline (as the rightmost bacterium) from centre.
test example 2: Herba Polygoni Capitati extract is anti-to helicobacter pylori type strain and clinical isolates strain bacterium is active to be detected
Basically the method for reference test example 1, the embodiment 2-18 of take respectively is extract obtained is reagent, test the antibacterial activity of these extracts to ATCC700392, ATCC700824, Hp B-1, Hp B-2, Hp B-3, Hp B-4, Hp B-5, the all extracts of result to the MIC value of these seven kinds of helicobacter pylori between 0.05 ~ 8mg/mL, the MIC value of seven kinds of helicobacter pylori that for example embodiment 2-10,17 extracts are right is between 1 ~ 8mg/mL, and the MIC value of seven kinds of helicobacter pylori that embodiment 11-16,18 extracts are right is between 0.05 ~ 2mg/mL.
test example 3: experiment in the body of Herba Polygoni Capitati extract (embodiment 1 is extract obtained)
By building the model of Helicobacter pylori infection mice, then, with after the Herba Polygoni Capitati treatment, observe the elimination situation of Hp and the improvement situation of stomach lining inflammation, the judgement Herba Polygoni Capitati suppresses or kills the effect of Hp in vivo.Experimental strain selection standard strains A TCC700392 and SS1, laboratory animal is selected SPF level BALB/c mouse.Experimental technique is as follows:
1) build the animal model grouping: type strain 700392 and SS1: negative control group, positive controls and experimental group.
2) recovery bacterial strain: recovery reference culture ATCC700392 and SS1, and go down to posterity once, be respectively to the animal gavage and prepare.
3) animal pretreatment: to animal marshalling numbering; Jejunitas, 24h runs out of rice and fuel; 50% ethanol 0.3ml gavage, continue jejunitas running out of rice and fuel.
4) infect Hp: after alcohol pre-treatment 24h with 10 9self-regulated bacterium liquid 0.5ml gavage; 0.5ml/ only upper and lower noon each 1 time, fill with and feed 1 time with dosage with same concentration the morning every other day, matched group is filled with and is fed with aseptic meat soup, for food after 4h, water.Infect after 8 weeks and make the animal grouping carry out administration.Get in addition Some Animals, after infecting, last puts to death respectively mice after the 4th week, the 8th week and the 12nd week, observe the mouse model success of whether building Helicobacter pylori infection, and whether infect sustainable existence, observation item is as follows: (1) takes out Mus gastric antrum section mucosal tissue after putting to death, and half is cultivated, smear and the test of the RUT scraps of paper; (2) animal eyeball blood-letting, separation of serum, preserve the ELISA test for-20 ℃ and detect anti-Hp antibody; (3) second half mucosal tissue dyes to observe the pathological changes such as degree of inflammation of its Hp field planting situation and mucosal tissue as HE.Result show to infect after 8 weeks all animals modeling success and still sustainable existence infection during by 12 weeks.
5) Drug therapy and detection: infect after 8 weeks and make the animal grouping carry out administration, every group of 20 animals.Experimental group gives embodiment 11 extracts, 250mg/kg/d (5% CMC-Na solution dissolving/suspendible), continuous 14 days; Positive controls gives amoxicillin, 250mg/kg/d (5% CMC-Na solution dissolving/suspendible), continuous 14 days), negative control group gives the CMC-Na solution of same volume.Whether 4 weeks observation mice stomach symptoms after treatment finishes, improve and whether helicobacter pylori is eradicated.Perusal, the result demonstration, in the test of every kind of bacterial strain, the gastric mucosa of animal in experimental group and positive controls more than 80% is normal, and without ulcer, the demonstration helicobacter pylori is eradicated, and negative control group all animals stomach all has obvious ulcer surface.
test example 4: Herba Polygoni Capitati extract infects the gastric mucosal protective effect of C57BL/6 mice to Hp
1, materials and methods
Bacterial strain: helicobacter pylori ATCC700392 international standard order-checking strain, purchased from ATCC (American Type Culture Collecti), passage number is within 10 generations.Helicobacter pylori Hp Sydney strain SS2000 is so kind as to give by infectious disease institute of Chinese Center for Disease Control (CDC) Zhang Jianzhong, professor He Lihua.Animal: 70 SPF level C57BL/6 mices, in 6 ~ 8 week age, ♀ 35,
Figure BDA00002167364700221
body weight (20 ± 5) g is provided the quality certification number by Medical University Of Chongqing's Experimental Animal Center: SYXK (Chongqing) 2007-0001.
Medicine: Herba Polygoni Capitati extract powder, brown ceramic powder, crude drug content is 8.72g/g, flavones content: 18.2mg/g, gallic acid content: 17.8mg/g, the powdered extract powder obtained according to the preparation method of " Pharmacopoeia of People's Republic of China 2010 version one one " " the clear granule of pyretic stranguria " kind, do not add adjuvant, below available code name HL-1112 mean this medicine.
During use, can be dissolved into concentrated solution (for example 320mg/mL) with aseptic double-distilled water, 4 ℃ save backup, and face with dilution.
2, experimental technique
(1) detection of Miao Ethnomedicine HL--1112 to Hp minimum inhibitory concentration (MIC)
The preparation of Colombia's blood agar plate: take Colombia blood agar powder 3.9g, add the 89mL deionized water, (121 ℃ of autoclavings, 20min) after processing, when agar is cooled to 50 ℃~55 ℃, adding the aseptic defiber Sanguis caprae seu ovis of 10mL (10%), 1mL (1%) to mix antibiotic fully palms off as, being poured into diameter is in the aseptic plate of 90mm, making agar thickness is 3mm~4mm, and get at random a ware and be placed in 37 ℃ of calorstats and cultivate 24h and do sterility test, it is standby that all the other all put into 4 ℃ of refrigerators.
The cultivation of helicobacter pylori SS2000 and ATCC 700392: take out bacterial strain to be measured-helicobacter pylori SS2000 and ATCC 700392 from-80 ℃ of refrigerators, room temperature is poured in Colombia's blood agar plate after dissolving, coating evenly, is cultivated 72h (5%O in 37 ℃ of calorstats 2, 10%CO 2, 85%N 2, relative humidity is more than 95%), observe the bacterial growth situation and carry out the Hp identification experiment: Gram’s staining, oxidase, catalase, urease experiment; By identification experiment all positive antibacterial gone down to posterity and cultivate after 72h repeated observation bacterial growth situation and reach and carry out the Bacteria Identification experiment, by antibacterial again transferred species cultivate 72h in Colombia's blood agar plate.
The configuration of HL-1112 medicine plate: take 17.55g Colombia solid agar powder and be dissolved in the 378mL deionized water, after being sub-packed in the sterilizing of 9 conical flask inner high voltages, when being cooled to 50~55 ℃ of left and right, culture medium add 5mL (10%) defiber Sanguis caprae seu ovis, 0.5mL (1%) to mix antibiotic, add again corresponding dilution medicinal liquid 2.5mL, after fully mixing, pour in 2 aseptic plates (90mm diameter), solidify rear 4 ℃ of preservations.The dilution factor of Miao Ethnomedicine HL-1112 solution and corresponding dull and stereotyped seeing the following form containing concentration.
Figure BDA00002167364700231
Drug sensitive experiment: the helicobacter pylori that will cultivate the 72h cell age heart infusion broth of requiring mental skill prepares bacteria suspension, allocates its concentration 3 * 10 8cFU/mL, aseptic cotton carrier picks bacteria suspension and evenly coats on the medicine culture dish and blank culture dish containing variable concentrations, and each drug level is smeared two plates, is placed in 37 ℃ of micro-aerobic environments and cultivates 72h.On blank culture dish, bacterial growth is good, in the situation that Miao Ethnomedicine HL-1112 is 4mg/mL to the MIC of Hp ATCC700392, the minimum drug level that has no bacterial growth of take on the pastille culture dish is the minimal inhibitory concentration (MIC) of Miao Ethnomedicine HL-1112 to Hp SS2000, and grown antibacterial is cooked to Gram’s staining and biochemical identification experiment.
(2) model of mice with helicobacter pylori infection chronic gastritis is set up
Helicobacter pylori is cultivated: take out the frozen Hp SS2000 bacterial strain at-80 ℃, after room temperature is dissolved, be poured on Colombia's blood agar culture-medium, after mixing, the plate of inoculation is put into to three gas incubator (5%O 2, 10%CO 2, 85%N 2), humidity keeps more than 90%, cultivates 72h, twice of continuous passage.
Bacterium solution preparation: the C57BL/6 adaptability is raised 1 week before modeling, after animal is divided into to normal group (20) and modeling group (50) by body weight, male and female stratified random; Hp cultivates the 72h results, the scraping bacterium colony, and in suspendible and aseptic brain heart infusion meat soup, ultraviolet spectrophotometer is measured bacterial concentration, and when A600=1, bacterial concentration is 2.2 * 10 8cfu/mL, by bacterial concentration modulation 1 * 10 9cfu/mL, 4 ℃ of preservations.
Hp Mice Inoculated: according to the Introduction of Literatures inoculated with Hp; Concrete steps are as follows: water 12h is prohibited in fasting, at first to mice, fills with and raises 0.3mL 50% ethanol, the Hp reference culture SS2000 bacterium liquid 0.5mL (1 * 10 that inoculation prepares every other day 9cfu/mL), interval 1d, gavage is 5 times altogether, and normal group replaces with the aseptic brain heart infusion meat soup of equivalent; All mouse stomaches are processed front fasting, are prohibited water 12h, fasting after gavage, taboo water 4h; After last gavage 8w, fasting, fasting 24h, randomly draw 10 mices from normal group and modeling group, after execution, dissect immediately, take out gastral cavity, cut off along the greater gastric curvature side, aseptic PBS cleans and rinses gastric content, vertically be divided into 3 parts, wherein 1 block organization's row RUT detects, and carries out the capable Hp of doubling dilution after the homogenate of 1 block organization and cultivates, and another 1 block organization is through paraffin embedding, after section, row HE and Giemsa dyeing, carry out histopathology and Hp and check; Specimen has typical characteristic and the equal positive of urease experiment, catalase test and oxidase test to suspicious bacterium colony Gram’s staining after cultivating and is judged to be Hp and infects; Whether the histopathology judgement, according to the diagnostic criteria of chronic gastritis Sydney, adopts the sxemiquantitative point system to be marked to Hp density, chronic inflammatory disease, course inflammatory activity, atrophy and intestinal epithelial metaplasia, successful with the inflammation modeling.
Sample disposal:
(a) Hp field planting standard: RUT experiment, fresh Mouse Gastric Mucous Membrane tissue is put into to urease reagent, tissue is put into rear 30min reagent, and to become red or aubergine person by orange colour be the positive.Giemsa dyeing, paraffin section enters in dimethylbenzene to dewax, ethanol gradient dehydrations at different levels, the 30min that dyes in the 2%Giemsa dyeing liquor, dimethylbenzene is transparent, mounting, the Hp thalline is navy blue.The tissue culture of Hp: after the weighing mucosal tissue, homogenate, get 200 μ L evenly coating and blank dull and stereotyped cultivation of Colombia's blood agar, after cultivating 5 ~ 7d, the capable urease of bacterium colony, catalase and the oxidase of turning out carried out to the Hp evaluation, and smear for microscopic examination Hp form.In the three, two positives are the Hp field planting positive.
(b) Hp plantation density: after the weighing mucosal tissue, homogenate, then go homogenate to do 10 -1serial dilution, each dilution factor is got 200 μ L evenly coating and blank dull and stereotyped cultivation of Colombia's blood agar, after cultivating 5 ~ 7d, and the viable count in the counting flat board, then be converted into the viable count (cfu) in every gram gastric tissue, plantation density means with the Igcfu/g tissue;
(c) Mouse Stomach histopathology: piece of tissue is conventional fixing with 10% formaldehyde, ethanol dehydration, and dimethylbenzene is transparent, paraffin embedding, 4 μ m sections.60 ℃ of roasting sheet 2h.HE dyeing: tissue slice, through dewaxing and the dehydration of gradient Ethanol Treatment, dyes in haematoxylin dyeing liquid, hydrochloride alcohol color separation, the color separation of ammonia ethanol, dyeing 5min in Yihong after rinsing, conventional dehydration, transparent, mounting.Karyon is dyed blueness, kytoplasm and Hp levelling pinken.
(d) criterion of pathological change: the gastritis of mice is with reference to the classification method of people's gastritis (2000 sorting technique in national chronic gastritis seminar National Consensus), and the degree of depth that infiltrates coat of the stomach according to the order of magnitude of chronic inflammatory cells (lymphocyte, plasma cell and eosinophilic granulocyte) is divided into 3 grades and score: NIP 0 minute; Inflammation is confined to mucosa shallow-layer 1/3 and thinks for slightly counting 1 minute; Deeply reaching 2/3 is moderate meter 2 minutes; Surpass 2/3 or holostrome the intensive person of cell be severe meter 3 minutes.The course inflammatory activity score: lamina propria has the neutrophilic granulocyte meter that is dispersed in 1 minute, and lamina propria has more neutrophilic granulocyte meter 2 minutes, and lamina propria has a large amount of neutrophilic granulocyte meters 3 minutes.
(e) modeling success criterion: have Hp field planting and gastric mucosa to occur that the chronic inflammatory disease pathological change is judged as the modeling success in Mouse Stomach.
(3) Miao Ethnomedicine HL-1112 therapeutic intervention Hp infecting mouse chronic gastritis
(a) mice group: modeling group mice is divided into to 4 groups, 10 every group, is respectively dosage group and HL-1112 low dose group in model group, HL-1112 high dose group, HL-1112.
(b) dosage regimen: Miao Ethnomedicine HL-1112 extractum, crude drug content is that 8.72g/g gets, can't measure its LD50, Miao Ethnomedicine HL-1112 to the maximum tolerated dose of mice be its 1/5MTD of 249.0g/kg/d, 1/10MTD, 1/20MTD respectively as high, medium and low group of dosage, be made into the high dose that concentration is 49.8g/kg/d, the middle dosage of 24.9g/kg/d and the low dosage of 12.45g/kg/d with distilled water.Start gastric infusion after the modeling success, model group all gives aseptic PBS 0.5mL, and every of the high, medium and low dosage group of Miao Ethnomedicine HL-1112 gives respectively the HL-1112 aqueous solution 0.5mL of high, medium and low dosage.Below respectively organize equal every day of gavage 1 time, successive administration 2w.Blank group: the normal raising, not gavage.
(c) observe the curative effect: 4w after treatment finishes, put to death experiment mice cervical vertebra dislocation method the taking-up gastric tissue of cutting open the belly, along the greater gastric curvature stringer, cut open, remove gastric content with sterilizing PBS washing, get gastric antrum section lesser curvature side tissue, vertically be divided into 3 parts, wherein 1 block organization's row RUT detects, carry out the capable Hp of doubling dilution after the homogenate of 1 block organization and cultivate, it is fixing that another 1 block organization is placed in 10% formalin, paraffin embedding, after section, row HE and Giemsa dyeing, carry out histopathology and Hp and check.Before mice is put to death, water 24h is prohibited in fasting.
(d) statistical analysis: measurement data means with " mean value ± standard deviation ", uses SPSS 15.0 software kits to carry out statistical analysis.Adopt each group difference of F inspection statistics, relatively adopt in twos the q check.
3, experimental result
(1) cultivation of Hp bacterial strain and evaluation
After Hp international standard strain SS2000 cultivates 72h on Colombia's blood agar plate, the tiny tip-like, smooth moistening, translucent glossy that is of bacterium colony.Microscopy after the smear Gram’s staining, the G-bacillus that antibacterial is helical form, S shape, arc or sea-gull shape, biochemical identification experiment urease, catalase, oxidase are all positive.
(2) detection of Miao Ethnomedicine HL-1112 to Hp minimum inhibitory concentration (MIC)
Good with bacterial growth in blank plate, under the contrast of Miao Ethnomedicine HL-1112 to the minimum inhibitory concentration MIC=4mg/mL of Hp international standard order-checking strain ATCC 700392, experiment repeats 3 times, all observe Hp growth conditions in the pastille plate of 0.125mg/mL, 0.5mg/mL, 1mg/mL good, in the pastille plate of 2mg/mL the growth thin, in the plate of 4mg/mL, 8mg/mL and 16mg/mL there are no bacterial growth.Therefore Miao Ethnomedicine HL-1112 is 4mg/mL to the minimum inhibitory concentration MIC of Hp SS2000.
(3) Hp field planting and distribution
The urease experiment of normal group animal gastric antrum, body of stomach and duodenal mucosa, Giemsa dyeing and gastric mucosa Hp cultivation results all show feminine gender.After inoculated with Hp during 8w, model group mice gastric antrum, gastric body mucosa urease-positive rate, Giemsa dyeing show that field planting rate, Hp cultivate positive rate and be 100%, Giemsa dyeing is presented at the field planting that body of stomach and gastric antrum section all can be observed antibacterial, the field planting antibacterial observed with the mucomembranous gland at gastric antrum position is maximum, also visible a large amount of antibacterials in gastric gland surface and upper strata mucus.After HL-1112 intervenes, its high, medium and low dosage group mice histopathology Giemsa dyeing, major part has no the Hp field planting and organizes at gastric antrum.
(4) mice histopathologic examination result
The pathological change of the gastric mucosa of model group mice appearance and the similar chronic active gastritis of the mankind after last bacterium liquid gavage 8w, inflammatory cell infiltration appears, integrity and the seriality of gastric mucosa are damaged, reach the visible a large amount of lymphocytic infiltrations of lamina propria between the gastric epithelial cell centered by the pylorus mucosa, centre is mingled with a small amount of neutrophil infiltration and plasmocyte infiltrating, and the inflammation scoring is 2.5 minutes.The gastric mucosa of blank group mice is complete, the body of gland queueing discipline, and interstitial, mucosa lamination show no obvious abnormalities, rare lymphocytic infiltration, pathological score is 0.5 minute.By statistics, model group compares with blank group, and the infiltration degree difference of esogastritis sexual cell has significant.
HL-1112 low dose group Mouse Gastric Mucous Membrane is 1 ~ 1.5 minute around the cell infiltration be dispersed on a small quantity, pathological score being arranged around muscular layer of mucosa and body of gland as seen; The middle and high dosage treatment group of HL-1112 Mouse Gastric Mucous Membrane pathological changes, be shown in the inflammatory cell that minute quantity is arranged between mucous layer and body of gland, and pathological score is 1 minute.The mice chronic gastritis that visible HL-1112 causes helicobacter pylori infections is significantly improved effect.In the other test, with reference to said method, give amoxicillin or omeprazole (the latter two dosage are respectively 10mg/kg/d, 1mg/kg/d) when giving HL-1112 simultaneously, result shows than the respective sets of alone HL-1112 compares pathological score all low 0.5 minute, shows that two class drug regimen results of use are better.
(5) respectively organize the Hp eradication rate: the result demonstration, the high, medium and low dosage group of HL-1112 and model group more all have significant difference (P<0.05); The high, medium and low dosage group of HL-1112 and the more equal no difference of science of statistics of blank group (P > 0.05), not statistically significant (P > 0.05) relatively in twos between the high, medium and low dosage group of HL-1112, illustrate between the high, medium and low dosage group of HL-1112 relatively the eradication rate no significant difference to Hp.

Claims (10)

1. Herba Polygoni Capitati or the purposes of its extract in the medicine for the preparation for the treatment of and/or preventing the disease relevant with Helicobacter pylori infection or disease.
2. the purposes of claim 1, wherein said Herba Polygoni Capitati is fresh goods or the dry product of Herba Polygoni Capitati.
3. the purposes of claim 1, wherein said Herba Polygoni Capitati is herb, aerial parts, leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
4. the purposes of claim 1, wherein said Herba Polygoni Capitati is the aerial parts of Herba Polygoni Capitati.
5. the purposes of claim 1, wherein said Herba Polygoni Capitati is leaf, flower, stem, seed or its combination that is selected from Herba Polygoni Capitati.
6. the purposes of claim 1, wherein said extract is that Herba Polygoni Capitati water, ethanol, ethanol water or its combination are as solvent, through extracting the extract obtained; Or Herba Polygoni Capitati is through the extract of carbon dioxide supercritical fluid extraction method acquisition.
7. the purposes of claim 6, wherein said ethanol water is 30~99% ethanol waters.
8. the purposes of claim 1, wherein said extract is the extract prepared according to the method comprised the steps: decocted and/or reflux, extract, concentrated extracting solution, drying by medicinal material of polygonum capilalum water, ethanol or ethanol water.
9. the purposes of claim 1, wherein said extract is the extract prepared according to the method comprised the steps: get medicinal material of polygonum capilalum, add the water, ethanol of 5~20 times or ethanol water decocts and/or reflux, extract, 1~5 time, each 0.5~5 hour, collecting decoction, filter, and filtrate decompression is concentrated into the thick paste that relative density is 1.1~1.5 (25 ℃), drying, obtain.
10. the purposes of claim 1 to 9 any one, the wherein said disease relevant with Helicobacter pylori infection or disease are selected from: gastric mucosa dependency lymphoma, peptic ulcer, gastric ulcer, duodenal ulcer, acute or chronic gastritis, superficial gastritis, atrophic gastritis, gastric cancer, cardiovascular disease, coronary heart disease, arteriosclerosis, cerebral infarction, hypertension, anemia, iron deficiency anemia, recurrent peptic ulcer, Mucosal atrophy, intestinal epithelial metaplasia and thrombocytopenic purpura, and their complication.
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