CN103599097A - Application of Artoxanthochromane in helicobacter pyloridis-resistant drug - Google Patents

Abstract

The invention discloses application of Artoxanthochromane in preparation of a helicobacter pyloridis-resistant drug. Artoxanthochromane can be used for treating acute gastritis, chronic gastritis, gastric ulcer, duodenum ulcer and other diseases and is applied to the preparation of drugs for treating acute gastritis, chronic gastritis, gastric ulcer and duodenum ulcer. Artoxanthochromane has a brand new type of skeleton, has high inhibitory activity for helicobacter pylori as well as outstanding substantial characteristics, and also shows a remarkable progress on prevention and treatment of helicobacter pyloridis infection.

Description

The application of Artoxanthochromane in Anti-helicobacter pylori drugs

Technical field

The present invention relates to the new purposes of compd A rtoxanthochromane, relate in particular to the application of Artoxanthochromane in preparing Anti-helicobacter pylori drugs.

Background technology

Helicobacter pylori (Helicobacter pylori, Hp) is a kind of Gram-negative spiral bacteria.Studies show that, helicobacter pylori is the main pathogenesis of acute and chronic gastritis and Stomach duodenum ulcer, and may be relevant with the morbidity of gastric cancer stomach function regulating mucosa-associated lymphoid tissue (MALT) malignant lymphoma.Recently, World Health Organization (WHO) is classified as I class carcinogen by Hp, and it plays a leading role in gastric cancer development.The scheme that at present popular treatment Hp infects is to take the triple therapy that proton pump inhibitor (PPI) adds two kinds of antibiotic (clarithromycin, amoxicillin, tetracycline, metronidazole etc. select two kinds) simultaneously.The main factor that affects 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 cause 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 compd A rtoxanthochromane the present invention relates to is one and within 2013, delivers (Horng-Huey Ko, et al., A Novel MonoterpeneStilbene Adduct with a4, 4-Dimethyl-2, 3-diphenylchromane Skeleton from Artocarpus xanthocarpus.chemistry & biodiversity, 2013 (10): noval chemical compound 1269-1275.), this compound has brand-new framework types, current purposes finds that it can remove oxygen-derived free radicals (Horng-Huey Ko, et al., A Novel MonoterpeneStilbene Adduct with a4, 4-Dimethyl-2, 3-diphenylchromane Skeleton from Artocarpus xanthocarpus.chemistry & biodiversity, 2013 (10): 1269-1275.), it is open first that the purposes of the Artoxanthochromane the present invention relates in preparing Anti-helicobacter pylori drugs belongs to.

Summary of the invention

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

Described compd A rtoxanthochromane, structure is as shown in formula I:

The experiment in vitro of Artoxanthochromane shows, Artoxanthochromane has very strong anti Helicobacter pylori activity, and paper disk method shows that its antibacterial circle diameter is 17mm (ATCC43504).With agar dilution, show that it can suppress the growth of 5 random clinical strains (Hp001, Hp003, Hp004, Hp018 and Hp036) and 1 reference culture (ATCC43504) completely, minimal inhibitory concentration (MIC) is 0.43 μ g/ml.With ampicillin, make positive control, its Cmin (MIC) that 6 strain test bacterium are suppressed is completely 3.9 μ g/ml.

This result of study shows, the energy force rate ampicillin of the inhibition helicobacter pylori activity of Artoxanthochromane is strong, explanation for the diseases such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, Artoxanthochromane 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 Artoxanthochromane the present invention relates in preparation treatment Anti-helicobacter pylori drugs belongs to open first, because framework types belongs to brand-new framework types, and it suppresses active strong for helicobacter pylori, possess outstanding substantive distinguishing features, the control for helicobacter pylori infections simultaneously obviously has significant progress.

The specific embodiment

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

The preparation method of compd A rtoxanthochromane involved in the present invention is referring to document (Horng-Huey Ko, et al., A Novel MonoterpeneStilbene Adduct with a4,4-Dimethyl-2,3-diphenylchromane Skeleton from Artocarpus xanthocarpus.chemistry & biodiversity, 2013 (10): 1269-1275.), prepare according to the method described above compd A rtoxanthochromane.

Embodiment 1: the preparation of compd A rtoxanthochromane tablet involved in the present invention:

Get 5 and digest compound Artoxanthochromane, add 195 grams, dextrin, mix, conventional tabletting is made 1000.

Embodiment 2: the preparation of compd A rtoxanthochromane capsule involved in the present invention:

Get 5 and digest compound Artoxanthochromane, add 195 grams of starch, mix, encapsulatedly make 1000.

Below by pharmacodynamic experiment, further illustrate its pharmaceutically active.

The pharmacological evaluation of experimental example 1:Artoxanthochromane

1) strains tested

Helicobacter pylori reference culture ATCC43504 is purchased from U.S.'s strain and preserves center (American Type Culture Collection, 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; Patient to peptic ulcer, duodenal bulbar inflammation or gastritis verrucosa in continuous gastroscopy, first through RUT experiment, be defined as Hp positive, get again antral gastric mucosa 1-2 piece, after chopping, be inoculated in the Columbia selectivity agar culture medium containing 8% horse serum, trimethoprim 1.25g/L, Polymyxin B2 500U/L, vancomycin 10mg/L, in 37 ℃, under micro-oxygen environment, (5%O2,10%CO2 and 85%N2) cultivates 72 hours.Collect antibacterial, through smear Gram’s staining, oxidase, catalase and urease are accredited as after the positive, the pure culture of going down to posterity, and 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 can produce the needed micro-aerobic environment of Hp by chemical reaction.

3) biological activity determination

Adopt paper disk method to measure the inhibitory action of compound to helicobacter pylori, with agar dilution, measure the minimum inhibitory concentration of test sample.

I. paper disk method experiment

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

(B) switching experimental bacteria (be coated with bacterium) with microscale sampler get diluted 10 ⁸cFU/ml (1OD660=10 ⁸cFU/ml) the bacteria suspension 0.1ml of Hp spreads upon suitable culture dish surface equably.Be inverted in 37 ℃ of drying bakers and take out after 15min, object makes agar surface dry, standby.

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

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

(E) survey antibacterial circle diameter and take out after flat board, measure respectively each scraps of paper size of antibacterial circle diameter around.With reference to the result of matched group, can draw the result of testing sample sensitive experiment.In triplicate.

II. agar dilution is measured MIC

(A) first the preparation of medicine flat board becomes dimethyl sulfoxide for compound (DMSO) solution preparation of test the mother solution of 0.5mg/ml, 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, the concentration of DMSO in medium is less than 1%.The test compounds solution that 1ml is prepared separately adds 1ml in the 9ml Colombia culture medium of 50 ℃ and is incubated in the horse serum of 50 ℃ and fully mixes with being incubated, and casts in culture dish cooling.

(B) switching experimental bacteria (be coated with bacterium) with microscale sampler draw diluted 1 * 10 ⁸the bacteria suspension 0.1ml of CFU/ml Hp spreads upon culture dish surface equably, is inverted in 37 ℃ of drying bakers and takes out after 15min, and object makes agar surface dry, standby.

(C) determine that MIC (contains culture dish to be measured: 85%N2,10%CO at micro-aerobic bag ₂and 5%O ₂) in, insulation 37oC cultivates 72 hours, observes Hp growing state, with blank group contrast, take and there is no the sample of bacteria growing least concentration completely as minimum inhibitory concentration value.Positive control is ampicillin.

3, the pharmacological results of Artoxanthochromane

Experiment in vitro shows, Artoxanthochromane has very strong anti Helicobacter pylori activity, and paper disk method shows that its antibacterial circle diameter is 17mm (ATCC43504).With agar dilution, show that it can suppress the growth of 5 random clinical strains (Hp001, Hp003, Hp004, Hp018 and Hp036) and 1 reference culture (ATCC43504) completely, minimal inhibitory concentration (MIC) is 0.43 μ g/ml.With ampicillin, make positive control, its Cmin (MIC) that 6 strain test bacterium are suppressed is completely 3.9 μ g/ml.

It is strong that conclusion: Artoxanthochromane suppresses the energy force rate ampicillin of helicobacter pylori activity, explanation for the diseases such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, Artoxanthochromane 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.Artoxanthochromane in Anti-helicobacter pylori drugs, described compd A rtoxanthochromane structure is as shown in formula I:

   

   Formula I.