Anti-helicobacter pylori natural product composition and application thereof

Abstract

The invention belongs to the technical field of application of natural products, and particularly relates to an anti-helicobacter pylori natural product composition and application, the anti-helicobacter pylori natural product composition comprises raspberry extract and cinnamon extract, and the composition contains anti-helicobacter pylori active substances. Besides, the natural product composition is combined with a clinical standard treatment method for treating helicobacter pylori, so that the curative effect can be further improved, and the gastrointestinal motility can be enhanced by adjusting intestinal flora disorder caused by pylorus infection, reducing toxic and side effects of antibiotic treatment, protecting gastric mucosa and enhancing gastrointestinal motility. The composition consists of two natural medicinal and edible plant extracts, is suitable for long-term use and has few adverse reactions. The Lycojaponicumin C can be used for preparing anti-helicobacter pylori medicines and preventive health-care foods, has obvious significance in prevention and treatment of helicobacter pylori infection, and has a good development prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of the application of natural products, and particularly relates to a natural product composition for anti-Helicobacter pylori and its application. Background Art

[0002] Helicobacter pylori (Hp) is a microaerophilic Gram-negative spiral bacterium colonizing on the surface of gastric mucosal epithelium. Currently, about half of the people in the world are infected with Helicobacter pylori. Research shows that Helicobacter pylori infection often leads to digestive system diseases, including chronic gastritis and peptic ulcer, etc. In addition, it may even lead to the occurrence of gastric cancer and mucosa-associated lymphoid tissue (MALT) malignant lymphoma. Research shows that eradicating the gastric infection of Helicobacter pylori helps to treat peptic ulcer and can significantly reduce the incidence of gastric cancer. Currently, most treatments for Helicobacter pylori infection are based on the use of antibiotics, including standard triple therapy (a combination of a proton pump inhibitor (PPI) + two broad-spectrum antibiotics) and quadruple therapy (a combination of bismuth agent and the former), which have achieved good treatment effects. However, in recent years, due to the abuse of antibiotics in clinical practice, drug-resistant strains have emerged, resulting in a significant reduction in the efficacy of antibiotics.

[0003] Finding highly effective, safe, and low-toxic anti-Helicobacter pylori active drugs has become an urgent problem to be solved. China has a huge natural Chinese herbal medicine resource. In recent years, research shows that many Chinese herbal medicines have a direct inhibitory effect on Helicobacter pylori. Currently, the existing anti-Helicobacter pylori natural products generally have problems such as slow curative effect on Helicobacter pylori-induced gastric diseases, long treatment cycle, low efficiency, complex formula, and difficult preparation. Therefore, it is of great significance to develop a traditional Chinese medicine composition that can effectively treat Helicobacter pylori. Summary of the Invention

[0004] To solve the above problems, the present invention provides a traditional Chinese medicine composition for anti-Helicobacter pylori. This natural product composition has excellent anti-Helicobacter pylori effect and high biological safety. And it has the characteristics of simple preparation, can directly reach the lesion, short cure time, and not easy to relapse after cure.

[0005] The inventor of the present invention found through research that by jointly using two natural products that can regulate the cellular immunity of the body, namely raspberry and cinnamon, or extracts of the two, the prevention and treatment effect on Helicobacter pylori can be significantly improved.

[0006] The specific technical solution of the present invention is as follows: A composition, the raw materials of the composition include: raspberry extract and cinnamon extract.

[0007] The raspberry extract is obtained by subjecting raspberry fruits to three - time reflux extraction with 30 - 50 vol% ethanol, centrifuging to obtain the supernatant, and then performing vacuum concentration and drying. The cinnamon extract is obtained by subjecting cinnamon bark to three - time reflux extraction with 40 - 70 vol% ethanol, separating, performing reduced - pressure concentration, and spray - drying.

[0008] Cinnamon (Cinnamomum cassia Presl) is a medium - sized tree, belonging to the class of dicotyledons, the order of Laurales, and the genus of Cinnamomum. The main medicinal components of cinnamon are cinnamaldehyde, cinnamyl acetate, ethyl cinnamate, benzyl benzoate, benzaldehyde, coumarin, β - cadinene, calamenene, β - elemene, protocatechuic acid, trans cinnamic acid, cinnamaldehyde cyclic glycerol - 1,3 - acetal (9,2'-cis), Cassio side, cinnamoside, and cinnamanAX and other compounds. Research shows that cinnamon has the functions of supplementing primordial yang, warming the spleen and stomach, and benefiting the five internal organs, and has extremely strong antibacterial activity, and has good curative effects on various digestive tract diseases such as chronic gastritis, duodenal ulcer, and gastric ulcer.

[0009] Raspberry (Rubus idaeus L.) is derived from the dried fruit of Rubus chingii Hu. It is warm in nature, sweet and sour in taste, and belongs to the liver and kidney meridians. The organic acids, polysaccharides, diterpenes, triterpenes, flavonoids, alkaloids, phenolic acids, etc. contained in it all have extremely strong antibacterial, anti - inflammatory, antioxidant, and anti - tumor activities. At the same time, it can scavenge free radicals, promote platelet aggregation, induce apoptosis, and prevent fat accumulation. It can enhance the body's immunity, regulate endocrine function, and increase physical fitness.

[0010] Preferably, the mass ratio of the raspberry extract to the cinnamon extract is (1 - 5):(3 - 7).

[0011] Preferably, the mass ratio of the raspberry extract to the cinnamon extract is (3 - 5):(5 - 7).

[0012] In the present invention, the raspberry extract is obtained by subjecting raspberries to three - time reflux extraction with 30 - 50 vol% ethanol, centrifuging to obtain the supernatant, and then performing vacuum concentration and drying; the cinnamon extract is obtained by subjecting cinnamon to three - time reflux extraction with 40 - 70 vol% ethanol, separating, performing reduced - pressure concentration, and spray - drying.

[0013] In the present invention, both the raspberry extract and the cinnamon extract are powders.

[0014] The present invention also provides the use of the composition according to the above technical solution in the preparation of a medicament for anti-Helicobacter pylori.

[0015] The present invention also provides a medicament for anti-Helicobacter pylori, comprising the composition according to the above technical solution and pharmaceutically acceptable excipients.

[0016] Preferably, the pharmaceutically acceptable excipients include, but are not limited to, one or more of natural, semi-natural and fully synthetic flavoring agents, fillers, flavoring agents and disintegrants.

[0017] Preferably, the dosage form of the medicament includes, but is not limited to, tablets, powders or capsules.

[0018] In the present invention, the pharmaceutical powder can be prepared by the following steps: mixing cinnamon (extract) and raspberry (extract), adding excipients to obtain a powder material. Granulating, drying, packaging, labeling, and warehousing after passing the inspection.

[0019] In the present invention, the pharmaceutical tablets can be prepared by the following steps: mixing cinnamaldehyde and its extract and raspberry (extract), adding excipients to obtain a powder material, tabletting with a tabletting machine, packaging, labeling, and warehousing after passing the inspection.

[0020] The inventor of the present invention combines the natural products raspberry and cinnamon or their extracts, and obtains a remarkable effect of preventing Helicobacter pylori infection. Moreover, the composition of the present invention is a natural product and its extract, which has unique action mechanisms, small drug resistance, safety, good curative effect and other advantages. Description of the Drawings

[0021] Figure 1 It shows that under the conditions of raspberry extract:cinnamon extract = (1-5):(3-7) and raspberry extract:cinnamon extract = (3-5):(5-7), it has good anti-Helicobacter pylori ability. In the figure, A: raspberry extract:cinnamon extract = (1-5):(3-7) (taking 1:3 as an example); B: raspberry extract:cinnamon extract = (3-5):(5-7) (taking 3:5 as an example).

[0022] Figure 2 It shows that under different mass ratios, the composition has a better anti-Helicobacter pylori effect than a single natural product. In the figure, A: raspberry extract; B: cinnamon extract; C: raspberry extract:cinnamon extract = (1-5):(3-7) (taking 1:3 as an example); D: raspberry extract:cinnamon extract = (3-5):(5-7) (taking 3:5 as an example).

[0023] Figure 3 Illustrated is the minimum inhibitory concentration (MIC) of the composition against Helicobacter pylori at different mass ratios. In the figure, A: raspberry extract; B: cinnamon extract; C: raspberry extract:cinnamon extract = (1 - 5):(3 - 7) (taking 1:3 as an example); D: raspberry extract:cinnamon extract = (3 - 5):(5 - 7) (taking 3:5 as an example). Detailed implementation manners

[0024] The present invention provides a composition for resisting Helicobacter pylori and its application, which is used to solve the problems of high drug resistance and easy recurrence in the treatment of Helicobacter pylori by existing therapies.

[0025] The following examples are intended to illustrate the present invention rather than further limit the present invention. The present invention will be further elaborated in detail below with reference to the examples, but the present invention is not limited to these examples and the preparation methods used. Moreover, those skilled in the art can make equivalent substitutions, combinations, improvements or modifications to the present invention according to the description of the present invention, but these will all be included within the scope of the present invention.

[0026] Example 1 Extraction of raspberry extract The raspberry fruits were crushed and screened through a 40-mesh sieve to obtain raspberry powder.

[0027] An accurate amount of raspberry powder was weighed and 50 vol% ethanol was added according to a solid-liquid ratio of 1:38, and heated under reflux at 80 °C for 3 h. After the extraction, the extract was centrifuged at 5000 r / min for 10 min, and the supernatant was collected.

[0028] The obtained supernatant was concentrated and dried to obtain raspberry extract.

[0029] Example 2 Extraction of cinnamaldehyde extract The cinnamon bark was crushed and screened through sieves of (250 ± 9.9) μm and (180 ± 7.6) μm to obtain uniform medium cinnamon powder.

[0030] An appropriate amount of medium cinnamon powder was weighed and placed in a round-bottom flask. A 70% ethanol solution was added according to a liquid-solid ratio of 1:8 (mass ratio), and extracted for 30 min, heated under reflux at 80 °C for 3 h (starting from when the solution began to boil), repeated 2 times, filtered, and the filtrates were combined; concentrated under reduced pressure at 50 °C until there was no alcohol to obtain a water-containing residue, and spray-dried to obtain cinnamon extract powder.

[0031] Example 3 Determination of motility inhibition The Helicobacter pylori solution was diluted with brain heart infusion broth (BHI) medium, and the bacterial concentration was adjusted to 1.2×10 9CFU / mL. Using a disposable inoculation needle, insert the adjusted bacterial suspension into the semi-solid motility medium. For each concentration, use three plates and incubate at 37°C in a microaerophilic environment for 5 days and observe. It was found that no obvious colonies grew under this combination (Group A and Group B), indicating that this composition has an inhibitory effect on the growth of Helicobacter pylori. (A: Raspberry extract: Cinnamon extract = (1 - 5):(3 - 7) (taking 1:3 as an example); B: Raspberry extract: Cinnamon extract = (3 - 5):(5 - 7) (taking 3:5 as an example)). See Figure 1 。

[0032] Example 4 Determination of Inhibition Zone Collect the bacteria cultured in Columbia blood agar medium at 37°C in a microaerobic environment (5% O2, 10% CO2 and 85% N2) for 48 - 72 hours. After the bacteria cover the plate, collect the bacteria, smear and Gram stain them. After being identified as positive for oxidase, catalase and urease, subculture and pure culture are carried out, and the obtained strain is used as the experimental strain.

[0033] Use a 1 mL pipette tip to aspirate 200 μL of the diluted 10 8 CFU / mL (1 OD 660 = 10 8 CFU / mL) Helicobacter pylori suspension, and use a sterilized stainless steel spreader to evenly spread it on the surface of a sterilized Columbia solid culture dish. Allow the bacteria on the agar surface to dry completely.

[0034] Prepare antibiotic and sample liquid medicine paper discs. Dissolve 5 mg of metronidazole in 1 mL of sterilized water, filter it through a 0.22 μm filter head to obtain a 5 mg / mL stock solution, and perform two-fold dilution with high-pressure sterilized water to obtain a 128 μg / mL working solution. Dissolve 5 mg of each group of drugs in 1 mL of sterilized water, filter it through a 0.22 μm filter head to obtain a 5 mg / mL stock solution, and perform two-fold dilution to obtain sample liquid sterilized paper discs with different concentrations.

[0035] Use sterile forceps to pick up the prepared paper discs containing sample liquid medicine with different concentrations (two-fold dilution) (5 mg / mL - 19.53 μg / mL) to form different experimental groups. The negative control (sterile aqueous solution) and the positive control (0.025% metronidazole by mass concentration) are placed on the agar surface containing bacteria according to aseptic operation, with one paper disc attached every certain distance. Use the paper discs to measure the effect of inhibiting the growth of Helicobacter pylori at 48 h and 72 h, and evaluate the ability to inhibit the growth of Helicobacter pylori by the size of the inhibition zone. Each drug concentration is repeated three times. Figure 2The diameters of the inhibition zones of 625 μg / ml raspberry extract, 625 μg / ml cinnamon extract and their mixtures against Helicobacter pylori growth (A: raspberry extract; B: cinnamon extract; C: raspberry extract:cinnamon extract = (1 - 5):(3 - 7) (taking 1:3 as an example); D: raspberry extract:cinnamon extract = (3 - 5):(5 - 7)) (taking 3:5 as an example).

[0036] It can be seen from Figure 2 that Columbia blood agar medium has no inhibitory effect on Helicobacter pylori, while both raspberry extract and cinnamon extract have inhibitory effects on the growth of Helicobacter pylori. When cultured for 48 h, the diameter of the inhibition zone of raspberry extract:cinnamon extract = (1 - 5):(3 - 7) can reach more than 13.2 ± 1.19 mm, and the diameter of the inhibition zone of raspberry extract:cinnamon extract = (3 - 5):(5 - 7) can reach more than 14.9 ± 1.16 mm. When cultured for 72 h, the diameter of the inhibition zone of raspberry extract:cinnamon extract = (1 - 5):(3 - 7) can reach more than 15.3 ± 1.36 mm, and the diameter of the inhibition zone of raspberry extract:cinnamon extract = (3 - 5):(5 - 7) can reach more than 16.5 ± 0.92 mm; the antibacterial effect is obvious and can be applied to the preparation of Helicobacter pylori inhibitors.

[0037] Determination of the minimum inhibitory concentration (MIC) in Example 5 The minimum inhibitory concentrations of raspberry extract, cinnamon extract, raspberry extract:cinnamon (extract) = (1 - 5):(3 - 7); raspberry extract:cinnamon extract = (3 - 5):(5 - 7); raspberry extract:cinnamon extract = 4:6 against Helicobacter pylori were determined by the microdilution method in 96-well plates.

[0038] The Helicobacter pylori solution was diluted with brain heart infusion broth (BHI) medium, 100 μL per well, and the final concentration was 1×10 per well 6CFU. Each compound to be tested (the compounds in Example 2) was diluted with 100 μL BHI containing 5% fetal bovine serum and added to a 96-well plate, and co-cultured for 48 - 72 h at 37 °C in a microaerophilic environment (5% O2, 10% CO2 and 85% N2). The minimum inhibitory concentration was measured at 625 nm using a multifunctional microplate reader. The growth of Hp was observed, and compared with the blank group. The lowest concentration of the sample with no bacterial growth was taken as the minimum inhibitory concentration (MIC) value. The positive control was metronidazole (MIC = 128 μg / mL had been screened out). The experimental results are shown in Table 3. Raspberry extract:cinnamon extract = (1 - 5):(3 - 7); raspberry extract:cinnamon extract = (3 - 5):(5 - 7) showed good anti-Helicobacter pylori activity and all exhibited antibacterial activity, showing good activity between 39.0625 - 156.25 μg / mL, see Figure 3 , and further screening was carried out. Finally, the MICs were 89 - 125 μg / mL and 89 - 116 μg / mL respectively. Figure 3 :(A: Raspberry (extract); B: Cinnamon (extract); C: Raspberry extract:cinnamon extract = (1 - 5):(3 - 7) (taking 1:3 as an example); D: Raspberry extract:cinnamon extract = (3 - 5):(5 - 7) (taking 3:5 as an example)).

Claims

1. A natural product composition, characterized in that , The raw materials of the composition include: raspberry extract and cinnamon extract. The raspberry extract is obtained by refluxing raspberry fruits three times with 30 - 50 vol% ethanol, centrifuging to obtain the supernatant, and then concentrating and drying it under vacuum. The cinnamon extract is obtained by refluxing cinnamon bark three times with 40 - 70 vol% ethanol, separating, concentrating under reduced pressure, and spray drying. The mass ratio of the raspberry extract to the cinnamon extract is (1 - 5):(3 - 7).

2. The composition according to claim 1, wherein , The mass ratio of the raspberry extract to the cinnamon extract is (3 - 5):(5 - 7).

3. Use of the natural product composition according to claim 1 or 2 in the preparation of a medicament for anti - Helicobacter pylori.

4. A natural product preparation against Helicobacter pylori, characterized in that , Using the natural composition according to claim 1 or 2, the dosage forms include but are not limited to tablets, capsules, infusion preparations, granules, pills, oral liquids or powders.

5. A health food, characterized in that , Comprising the natural product composition according to claim 1 or 2.

6. A health food, characterized in that , Comprising the natural product composition according to claim 1 or 2 and food - acceptable excipients.

7. The food according to claim 5, characterized in that , The food - acceptable excipients include but are not limited to one or more of natural, semi - natural and fully synthetic flavoring agents, fillers, flavoring agents and disintegrants.

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