CN105250259A - Application of Kanshone C to preparation of medicine for resisting helicobacter pylori - Google Patents

Abstract

In vitro activity experiments indicate that Kanshone C has strong activity in resisting helicobacter pylori (Hp). Kanshone C can be applied to treatment of acute gastritis, chronic gastritis, gastric ulcers, duodenal ulcers and other diseases as well as preparation of medicine for treating acute gastritis, chronic gastritis, gastric ulcers and duodenal ulcers. The usage of Kanshone C in preparing the medicine for treating and resisting helicobacter pylori is disclosed for the first time. Due to the fact that the framework type is brand new and the activity of Kanshone C in inhibiting helicobacter pylori is too strong to imagine, it is impossible to give any revelation by other compounds; Kanshone C has outstanding substantive characteristics and apparently has remarkable progress when applied to prevention and treatment of helicobacter pylori infection.

Description

Kanshone C is preparing the application in Anti-helicobacter pylori drugs

Technical field

The present invention relates to the novelty teabag of compound K anshoneC, particularly relate to KanshoneC and preparing the application in Anti-helicobacter pylori drugs.

Background technology

Helicobacter pylori (Helicobacterpylori, Hp) is a kind of Gram-negative spiral bacteria.Research display, helicobacter pylori is the primary pathogenic event of acute and chronic gastritis and taste-blindness rate, and may fall ill relevant with gastric cancer stomach function regulating mucosa-associated lymphoid tissue (MALT) malignant lymphoma.Recently, Hp is classified as I class carcinogen by World Health Organization (WHO), and it plays a leading role in stomach cancer development.Simultaneously the scheme that popular at present treatment Hp infects takes the triple therapy that proton pump inhibitor (PPI) adds two kinds of antibiotic (clarithromycin, amoxicillin, tetracycline, metronidazole etc. select two kinds).The main factor affecting triple therapy is considered to the drug resistance of Hp to antibacterial; Another serious problems are that proton pump inhibitor can bring out dyspepsia, and a large amount of antibacterial then causes the serious destruction of flora in digestive tract.Therefore, find the active kind new medicine thing of efficient, safe anti-Hp and become an important and urgent task.

The compound K anshoneC that the present invention relates to is one and delivers (KishorL in 2014, etal., TotalSynthesisof (±)-NardoaristoloneBandItsAnalogues.OrganicLetters, 2014, 16, noval chemical compound 4252-4255.), this compound has brand-new framework types, current purposes is the myocardial cell (KishorL of protection neonate rat, etal., TotalSynthesisof (±)-NardoaristoloneBandItsAnalogues.OrganicLetters, 2014, 16, 4252-4255.), the KanshoneC that the present invention relates to is belonged to first public preparing the purposes in Anti-helicobacter pylori drugs, owing to belonging to brand-new structure type, and its activity for anti-helicobacter pylori is unexpectedly strong, there is not the possibility being provided any enlightenment by other compounds, possesses outstanding substantive distinguishing features, for anti-helicobacter pylori, obviously there is significant progress simultaneously.

Summary of the invention

The object of the invention is to not find that it has the present situation of the report of anti-helicobactor pylori activity according in existing KanshoneC research, provide KanshoneC and preparing the application in Anti-helicobacter pylori drugs.

Described compound K anshoneC, structure is as shown in formula I:

The experiment in vitro of KanshoneC shows, KanshoneC has very strong anti Helicobacter pylori activity, and it is 16mm (ATCC43504) that paper disk method shows its antibacterial circle diameter.With agar dilution display, it can suppress the growth of 5 random clinical strains (Hp001, Hp003, Hp004, Hp018 and Hp036) and 1 reference culture (ATCC43504) completely, and minimal inhibitory concentration (MIC) is 0.34 μ g/ml.Make positive control with ampicillin, its Cmin (MIC) suppressed completely 6 strain test bacterium is 4.1 μ g/ml.

This result of study shows, the energy force rate ampicillin of the suppression helicobacter pylori activity of KanshoneC is strong, illustrate for the diseases such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, KanshoneC is the compound of a great exploitation potential for its.It can be directly used in the treatment of corresponding disease and the preparation of related drugs.

The purposes of the KanshoneC that the present invention relates in preparation treatment Anti-helicobacter pylori drugs belongs to first public, because framework types belongs to brand-new framework types, and it is unexpectedly strong for helicobacter pylori inhibit activities, there is not the possibility being provided any enlightenment by other compounds, possess outstanding substantive distinguishing features, the control simultaneously for helicobacter pylori infections obviously has significant progress.

Detailed description of the invention

The preparation method of compound K anshoneC involved in the present invention is see document (FushuangLi, etal., 2013.KanshoneC, aNewFlavanol-FusedStilbeneGlycosidefromPolygonumcuspidat um.OrganicLetters3 (15), 674 – 677.)

The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not by any restriction of specific embodiment, but be limited by claim.

Embodiment 1: the preparation of compound K anshoneC tablet involved in the present invention:

Get 20 g of compound KanshoneC, add the customary adjuvant 180 grams preparing tablet, mixing, conventional tablet presses makes 1000.

Embodiment 2: the preparation of compound K anshoneC capsule involved in the present invention:

Get 20 g of compound KanshoneC, add prepare capsule customary adjuvant as starch 180 grams, mixing, encapsulatedly makes 1000.

Its pharmaceutically active is further illustrated below by pharmacodynamic experiment.

The pharmacological evaluation of KanshoneC

1) strains tested

Helicobacter pylori (Helicobacterpylori, Hp) reference culture ATCC43504 is purchased from U.S.'s Culture Collection (AmericanTypeCultureCollection, ATCC).15 strain Hp clinical strains are picked up from Jiangsu Prov. People's Hospital Gastroenterology dept., clinical laboratory of Jiangsu TCM Hospital and Nanjing Children's Hospital Dndoscope Laboratory and are accepted gastroscopic patient; To the patient of peptic ulcer, duodenal bulbar inflammation or gastritis verrucosa in continuous gastroscopy, first be defined as Hp positive through RUT experiment, get antral gastric mucosa 1-2 block again, be inoculated in containing 8% horse serum, trimethoprim (trimethropin after chopping, in the Columbia selectivity agar culture medium of TMP) 1.25g/L, polymyxin (polymyxin) B2500U/L, vancomycin (vancomycin) 10mg/L, in 37 DEG C under micro-oxygen environment (5%O2,10%CO2 and 85%N2) cultivate 72 hours.Collect antibacterial, through smear Gram’s staining, after oxidase, catalase and urease are accredited as the positive, pure culture of going down to posterity, obtained strains is as experimental strain.

2) strain culturing

We adopt micro-aerobic bag (purchased from Shanghai Medical Univ) to carry out the strain culturing of HP, and it produces the micro-aerobic environment required for Hp by chemical reaction.

3) biological activity determination

Paper disk method (Microbiologicalpapermethod) is adopted to measure the inhibitory action of compound to helicobacter pylori, the minimum inhibitory concentration (minimalinhibitoryconcentration, MIC) of test sample is measured with agar dilution.

I. paper disk method experiment

(A) culture medium is prepared by the Columbia culture medium for preparing after high pressure steam sterilization, be cooled to 50-60 DEG C, add 8% horse serum or Sheep Blood, mixing is poured in the culture dish of sterilizing, every ware 7-10ml, culture medium thickness is 1.5mm (sterile working).

(B) experimental bacteria of transferring (being coated with bacterium) gets 108CFU/ml (1OD660=108CFU/ml) Hp diluted bacteria suspension 0.1ml with microscale sampler spreads upon suitable culture dish surface equably.Be inverted in 37 DEG C of drying bakers and take out after 15min, object makes agar surface dry, for subsequent use.

(C) paste sample scraps of paper microscale sampler to get 6 μ l testing samples (mass concentration 2mg/ml) and inject on the round filter paper of sterilizing.With aseptic nipper tweezer containing the scraps of paper of sample and the blank scraps of paper of contrast, by sterile working respectively the scraps of paper be close to containing bacterio-agar surface, paste a piece of paper sheet at a certain distance.Often kind of bacterium is cooked 3 wares, and acquired results asks its meansigma methods.

(D) cultivate each plate is placed in micro-aerobic bag, sealing, opens gas generator, then is placed in 37 DEG C of incubators and cultivates 72h.

(E), after surveying antibacterial circle diameter taking-up flat board, the size of antibacterial circle diameter around each scraps of paper is measured respectively.With reference to the result of matched group, the result of testing sample sensitive experiment can be drawn.In triplicate.

II. agar dilution measures MIC

(A) first compound dimethyl sulfoxide (DMSO) solution preparation of test is become the mother solution of 0.5mg/ml by the preparation of Drug plates, then with sterilized water dilution, is finally made into 10.0,8.0,6.0,4.5,4.0,3.5,3.0,2.5,2.0,1.5,1.0, the concentration series of 0.5 and 0.25 μ g/ml, DMSO concentration is in media as well less than 1%.The test compounds solution prepared by 1ml separately adds in the 9ml Columbia medium of 50 DEG C the horse serum that 1ml is incubated in 50 DEG C and fully mixes with being incubated, and casts in culture dish to cool.

(B) experimental bacteria of transferring (being coated with bacterium) draws the 1 × 108CFU/mlHp diluted bacteria suspension 0.1ml with microscale sampler spreads upon culture dish surface equably, be inverted in 37 DEG C of drying bakers and take out after 15min, object makes agar surface dry, for subsequent use.

(C) determine that test dish (contains: 85%N2,10%CO at micro-aerobic bag by MIC ₂and 5%O ₂) in, be incubated 37 DEG C and cultivate 72 hours, observe Hp growing state, contrast with blank group, there is no the sample least concentration of bacteria growing completely for minimum inhibitory concentration (minimalinhibitoryconcentration, MIC) value.Positive control is ampicillin (Ampicillin).

3, the pharmacological results of KanshoneC

Experiment in vitro shows, KanshoneC has very strong anti Helicobacter pylori activity, and it is 16mm (ATCC43504) that paper disk method shows its antibacterial circle diameter.With agar dilution display, it can suppress the growth of 5 random clinical strains (Hp001, Hp003, Hp004, Hp018 and Hp036) and 1 reference culture (ATCC43504) completely, and minimal inhibitory concentration (MIC) is 0.34 μ g/ml.Make positive control with ampicillin, its Cmin (MIC) suppressed completely 6 strain test bacterium is 4.1 μ g/ml.

Conclusion: KanshoneC suppresses the energy force rate ampicillin of helicobacter pylori activity strong, illustrate for the diseases such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, KanshoneC is the compound of a great exploitation potential for its.It can be directly used in the treatment of corresponding disease and the preparation of related drugs.

Claims (1)

1. The application of 1.KanshoneC in Anti-helicobacter pylori drugs, described compound K anshoneC structure is as shown in formula I:

   Formula I.