Chromone and acorane-type sesquiterpene hybrid compounds with inhibitory activity on helicobacter pylori and application thereof

Abstract

The application discloses a chromone and acorane-type sesquiterpene hybrid compound with helicobacter pylori inhibiting activity and application thereof, wherein the chromone and acorane-type sesquiterpene hybrid compound has a structure shown in formula I: formula I. The application further discloses application of the chromone and acorane-type sesquiterpene hybrid compound in preparation of a product with helicobacter pylori inhibiting activity and in preparation of a medicine for treating and / or relieving a disease related to helicobacter pylori. The chromone and acorane-type sesquiterpene hybrid compound, whose structural formula is not disclosed, can also inhibit generation of helicobacter pylori; can be used in preparation of a product with helicobacter pylori inhibiting activity, and further used in preparation of a medicine for treating and / or relieving a disease related to helicobacter pylori.

Description

Technical Field

[0001] This invention belongs to the field of natural medicine technology, specifically relating to a chromone and calamus spirulane type sesquiterpene hybrid compound with inhibitory activity against Helicobacter pylori and its application. Background Technology

[0002] Helicobacter pylori ( Helicobacter pylori As a major pathogen causing chronic gastritis, peptic ulcers, and even gastric cancer, its treatment has long relied on triple or quadruple therapy, primarily using commonly used antibiotics such as clarithromycin, metronidazole, and amoxicillin. However, antibiotic overuse has led to a significant increase in drug resistance rates globally, with some regions exceeding warning levels, and the eradication rate of traditional therapies declining year by year. Simultaneously, the side effects of antibiotics, such as gut microbiota dysbiosis and liver and kidney toxicity, have prompted researchers to turn their attention to natural products, attempting to screen for safer drugs.

[0003] Agarwood belongs to the genus Aquilaria in the family Thymelaeaceae. Aquilaria Agarwood is the wood formed from the resin accumulated and secreted by plants of the genus *Gyrinops* after external damage. In the treasure trove of traditional Chinese medicine, agarwood is an extremely precious medicinal material. It is not only a top-grade spice but also a good medicine for regulating the spleen and stomach and treating stomach ailments. Agarwood is warm in nature, pungent and bitter in taste, and enters the spleen, stomach, and kidney meridians. It has the effects of promoting qi circulation and relieving pain, warming the middle jiao and stopping vomiting, and regulating qi and relieving asthma. The main characteristic components of agarwood are sesquiterpenes and 2-(2-phenylethyl)chromone compounds, which have anti-inflammatory, antibacterial, and antitumor pharmacological effects.

[0004] Currently, there are no reports in the prior art regarding compounds that have inhibitory activity against Helicobacter pylori isolated from Sri Lankan Wara agarwood. Therefore, this application aims to provide a chromone and calamus spirulane type sesquiterpene hybrid compound with inhibitory activity against Helicobacter pylori and its application. Summary of the Invention

[0005] The purpose of this invention is to provide a chromone and calamus spirulane-type sesquiterpene hybrid compound that has the activity of inhibiting Helicobacter pylori.

[0006] The present invention also aims to provide a method for preparing the above-mentioned chromone and calamus spirulane type sesquiterpene hybrid compound with the activity of inhibiting Helicobacter pylori.

[0007] The final object of the present invention is to provide the use of the above-mentioned chromone and calamus spirulane type sesquiterpene hybrid or the chromone and calamus spirulane type sesquiterpene hybrid obtained by the preparation method described above in the preparation of products having activity against Helicobacter pylori and in the preparation of medicaments having therapeutic and / or alleviating reactions related to Helicobacter pylori.

[0008] The first objective of this invention can be achieved by the following technical solution: a chromone and calamus spirulane-type sesquiterpene hybrid compound having activity inhibiting Helicobacter pylori, wherein the chromone and calamus spirulane-type sesquiterpene hybrid compound has the structure shown in Formula I:

[0009] Formula I.

[0010] The second objective of this invention can be achieved through the following technical solution: the preparation method of the above-mentioned chromone and calamus spirulane-type sesquiterpene hybrid compound with Helicobacter pylori inhibitory activity includes the following steps: (1) Different fractions were obtained by heating and reflux extraction, extraction and vacuum normal phase silica gel column separation of Sri Lankan Wara agarwood. Then, liquid chromatography-mass spectrometry analysis was performed to determine the fraction containing chromone and sesquiterpene impurities, which was called fraction a. (2) The fraction a was separated and eluted sequentially using a C18 reversed-phase silica gel column and a Sephadex LH-20 gel column to obtain a mixed system b; (3) The mixture b was eluted by semi-preparative high performance liquid chromatography to obtain a chromone and a strophane-type sesquiterpene hybrid with the structure shown in Formula I.

[0011] In the above-mentioned method for preparing the chromone and calamus spirulane-type sesquiterpene hybrid compound with inhibitory activity against Helicobacter pylori: Preferably, step (1) after heating and reflux extraction further includes removing the solvent to obtain an extract paste, mixing the extract paste with water to obtain a suspension, and then extracting the suspension.

[0012] Preferably, the solvent used in step (1) for heating and reflux extraction is ethanol.

[0013] Preferably, the solvents used for extraction in step (1) are petroleum ether, ethyl acetate and n-butanol, in sequence.

[0014] Preferably, in step (1), the reduced pressure normal phase silica gel column separation is performed by gradient elution using a mixed solvent of chloroform and methanol to obtain 11 fractions, denoted as Fr.1 ~ Fr.11, and fraction Fr.5 is taken as fraction a.

[0015] More preferably, the reduced pressure normal phase silica gel column separation in step (1) is performed by gradient elution with a mixed solvent of chloroform and methanol at a volume ratio of 200:1 to 5:1 (200:1, 100:1, 50:1, 40:1, 30:1, 20:1, 10:1, 5:1, V / V) to obtain 11 fractions, denoted as Fr.1 to Fr.11, and the Fr.5 fraction is taken as fraction a.

[0016] Preferably, in step (2), the separation and elution of the C18 reversed-phase silica gel column is performed by gradient elution of fraction a with a mixed solvent of methanol and water to obtain 15 fractions, denoted as Fr.5-1 to Fr.5-15. Fraction Fr.5-15 is then used for the next step of separation and elution with the Sephadex LH-20 gel column.

[0017] More preferably, in step (2), the separation and elution of the C18 reversed-phase silica gel column is performed by gradient elution of fraction a using a mixed solvent of methanol and water at a volume ratio of 3:7 to 1:0 (the volume percentage of methanol is 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%, respectively), to obtain 15 fractions, denoted as Fr.5-1 to Fr.5-15. Fraction Fr.5-15 is then used for the next step of separation and elution using the Sephadex LH-20 gel column.

[0018] Preferably, in step (2), the separation and elution of the Sephadex LH-20 gel column is performed by eluting Fr.5-15 with methanol to obtain 5 fractions, denoted as Fr.5-15-1 to Fr.5-15-5.

[0019] Preferably, the elution of the semi-preparative high performance liquid chromatography in step (3) is to elute the stream Fr.5-15-2 with a mixed solvent of acetonitrile and water at a volume ratio of 50:50 to obtain a chromone and a strophane-type sesquiterpene heteropolymer having the structure shown in Formula I.

[0020] As a preferred embodiment of the present invention, the method for preparing a chromone and calamus spirulane-type sesquiterpene hybrid compound with Helicobacter pylori inhibitory activity provided by the present invention includes the following steps: S1: After crushing the Sri Lankan Wara agarwood, it was extracted five times by heating and refluxing with 2.5L of ethanol to obtain the extract. The extracts were then combined and concentrated into an ethanol extract. S2: Prepare a suspension by mixing the ethanol extract with water at a volume ratio of 1:1, and extract it sequentially with petroleum ether, ethyl acetate and n-butanol. Then concentrate each extract to prepare an extract. S3: The ethyl acetate extract was passed through a normal-phase silica gel column under reduced pressure and eluted with chloroform-methanol (200:1~5:1, V / V; specifically 200:1, 100:1, 50:1, 40:1, 30:1, 20:1, 10:1, 5:1) to obtain 11 fractions, denoted as Fr.1 ~ Fr.11; S4: Liquid chromatography-mass spectrometry analysis of the Fr.1 ~ Fr.11 components revealed that Fr.5 contained a chromone polymer with a molecular weight between 500-700 Da; S5: The fraction Fr.5 was subjected to gradient elution with methanol-water (3:7~1:0, V / V, with methanol volume percentages of 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%) on an octadecylsilane-bonded silica column to obtain 15 fractions, denoted as Fr.5-1~Fr.5-15; S6: The fraction Fr.5-15 was eluted with methanol through a Sephadex LH-20 gel column to obtain 5 fractions, denoted as Fr.5-15-1 to Fr.5-15-5; S7: The fraction Fr.5-15-2 was prepared by semi-preparative high performance liquid chromatography to obtain a compound with a molecular weight between 500-700 Da of Formula I. The high-performance liquid chromatography (HPLC) conditions in step S7 are as follows: C18 column; acetonitrile / water, 50:50, V / V; flow rate 4.0 mL / min; UV detection wavelength 210 / 254 nm.

[0021] The third objective of the present invention can be achieved by the following technical solution: the application of the above-mentioned chromone and calamus spirulane type sesquiterpene hybrid or the chromone and calamus spirulane type sesquiterpene hybrid obtained by the above-mentioned preparation method in the preparation of products with Helicobacter pylori inhibitory activity.

[0022] The present invention also provides a product having activity of inhibiting Helicobacter pylori, comprising the aforementioned chromone and calamus spirulane type sesquiterpene hybrid or the chromone and calamus spirulane type sesquiterpene hybrid obtained by the aforementioned preparation method.

[0023] Preferably, the product is a drug.

[0024] Preferably, the product further includes pharmaceutically acceptable excipients.

[0025] Preferably, the dosage form of the drug is tablets, capsules, powder for injection, or suspension.

[0026] The present invention further provides the use of the above-mentioned chromone and calamus spirulane type sesquiterpene hybrid or the chromone and calamus spirulane type sesquiterpene hybrid obtained by the above-mentioned preparation method in the preparation of medicaments for treating and / or alleviating diseases related to Helicobacter pylori.

[0027] Preferably, the Helicobacter pylori-related diseases include one or more of chronic gastritis, atrophic gastritis, and gastric ulcer.

[0028] The present invention has the following beneficial effects: The chromone and calamus spirulane-type sesquiterpene hybrid provided by the present invention, which have the activity of inhibiting Helicobacter pylori, have an undisclosed structural formula. The inhibition of Helicobacter pylori production was evaluated by MIC and MBC assays. The chromone and calamus spirulane-type sesquiterpene hybrid provided by the present invention can inhibit the production of Helicobacter pylori. Therefore, it is shown that the chromone and calamus spirulane-type sesquiterpene hybrid provided by the present invention can be used to prepare products with the activity of inhibiting Helicobacter pylori, and further used to prepare pharmaceuticals for treating and / or alleviating the activity of Helicobacter pylori. Attached Figure Description

[0029] Figure 1 It is the compound of formula I prepared in Example 1. 1 The 1H NMR spectrum is shown, where the horizontal axis represents the chemical shift f1 (ppm) and the vertical axis represents the peak height.

[0030] Figure 2 It is the compound of formula I prepared in Example 1. 13 C NMR spectrum, where the horizontal axis represents chemical shift f1 (ppm) and the vertical axis represents peak height.

[0031] Figure 3 The image shows the HSQC spectrum of the compound with structure I prepared in Example 1, where the horizontal axis is the chemical shift f2 (ppm) of 1H and the vertical axis is the chemical shift f1 (ppm) of 13C.

[0032] Figure 4 It is the compound of formula I prepared in Example 1. 1 H- 1 The H COSY spectrum shows the chemical shift of hydrogen f2 (ppm) on the horizontal axis and the chemical shift of hydrogen f1 (ppm) on the vertical axis.

[0033] Figure 5 The image shows the HMBC spectrum of the compound with structure I prepared in Example 1, where the horizontal axis represents the chemical shift of hydrogen f2 (ppm) and the vertical axis represents the chemical shift of carbon f1 (ppm).

[0034] Figure 6 This is the ROESY spectrum of the compound with structure I prepared in Example 1, where the horizontal axis represents the chemical shift f2 (ppm) of one nucleus ¹H and the vertical axis represents the chemical shift f1 (ppm) of the other nucleus ¹H.

[0035] Figure 7 This is the HRESIMS spectrum of the compound with structure I prepared in Example 1, where the horizontal axis represents the mass-to-charge ratio of the ions (m / z) and the vertical axis represents the intensity of the ion current (abbreviated as Intens). Detailed Implementation

[0036] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0037] The implementation of the present invention will be described in detail below with reference to specific embodiments.

[0038] The chromone and calamus spirulane-type sesquiterpene hybrids with inhibitory activity against Helicobacter pylori provided in this invention can be implemented by those skilled in the art with appropriate modifications to the process parameters, based on the content of this document. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention are described through the following preferred embodiments. Those skilled in the art will clearly be able to modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to implement and apply the technology of this invention.

[0039] The experimental materials used in the following embodiments of the present invention are all commercially available products that can be purchased on the market.

[0040] Reference Figures 1-7 As shown, the following are preferred embodiments provided by the present invention.

[0041] The present invention provides a chromone and calamus spirulane-type sesquiterpene hybrid with inhibitory activity against Helicobacter pylori, a method for its preparation, and its application. The general structural formula of the chromone and calamus spirulane-type sesquiterpene hybrid is shown in Formula I:

[0042] Formula I.

[0043] The following is a specific implementation of the present invention: Example 1

[0044] 1.1 Recommendations for instruments, reagents, and materials, but not limited to: Bruker AV-500 superconducting nuclear magnetic resonance spectrometer (Bruker, Switzerland); Autospec 300 mass spectrometer (VG, UK); analytical high-performance liquid chromatograph (Agilent Technologies, USA); semi-preparative high-performance liquid chromatograph (Agilent Technologies, USA); N-1000 (2 L) vertical rotary evaporator and CA-1111 cooling water circulation system (Shanghai Ailang Instrument Co., Ltd.); SHZ-D (Ⅲ) circulating vacuum pump (Shanghai Longtuo Instrument Equipment Co., Ltd.); AS220.R2 0.001 g electronic balance (RADWAG Wagi Elektroniczne); Sephadex LH-20 gel (Merck Co., Ltd.); C18 reversed-phase silica gel (20~45µm, Fuji Silysia Chemical Ltd., Japan); C18 column (10.0 ID × 10.0 m³ / h). 250.0 mm, Nacalaitesque; silica gel for column chromatography and silica gel plates for thin-layer chromatography (Qingdao Ocean Chemical Plant); deuterated reagents and chromatographic methanol (Merck, Germany); commonly used organic reagents such as 95% ethanol, redistilled methanol, ethyl acetate, chloroform, petroleum ether, and acetone (Tianjin Kemeio, Tianjin Fuchen, Guangzhou Guanghua, etc.).

[0045] The Sri Lankan Wara agarwood sample (384.0g) was purchased in Bangkok, Thailand in August 2014. It was identified by Researcher Dai Haofu of the Institute of Tropical Biotechnology, Chinese Academy of Tropical Agricultural Sciences, as the original plant of Wara agarwood, belonging to the genus Gyrinopswalla Gaertn of the family Thymelaeaceae. The voucher specimen (No. 201408SLLK) is preserved at the Institute of Tropical Biotechnology, Chinese Academy of Tropical Agricultural Sciences.

[0046] 1.2 Preparation and structural identification of compounds Includes the following steps: (1) After the Sri Lankan Wara agarwood sample (384.0g) was crushed, it was extracted five times by heating (slight boiling) with 2.5L of ethanol aqueous solution with a volume percentage of 95%. The resulting extracts were filtered and combined, and concentrated under reduced pressure at 45℃ to obtain ethanol extract paste (129.4g). (2) Dilute the extract with 1.0L of water to form a suspension, and extract it three times in sequence with petroleum ether (1.0L each time), ethyl acetate (1.0L each time) and n-butanol (1.0L each time). The extract layers are evaporated and concentrated at 45°C to prepare petroleum ether phase extract, ethyl acetate phase extract and n-butanol phase extract, respectively. (3) Take 107.8 g of ethyl acetate extract and pass it through a normal-phase silica gel column (silica gel H) under reduced pressure and elute with a gradient of chloroform:methanol (200:1, 100:1, 50:1, 40:1, 30:1, 20:1, 10:1, 5:1, V / V). Collect 250 mL of each flask and use 5.0 L of mixed solution for each gradient. Detect each fraction by thin-layer chromatography (TLC). Combine similar fractions to obtain 11 fractions, which are denoted as Fr.1 ~ Fr.11. (4) Liquid chromatography-mass spectrometry analysis of the Fr.1 ~ Fr.11 components revealed that Fr.5 contained chromone and sesquiterpene heteropolymers with a molecular weight between 500-700 Da; (5) The fraction Fr.5 (3.2 g) was separated by C18 reversed-phase column chromatography (4.0 cm × 30.0 cm), eluted with methanol-water (methanol volume percentages of 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%), and collected in 100 mL bottles. Each gradient was mixed with 1.5 L of the mixture. The obtained fractions were detected by thin-layer chromatography (TLC) and similar fractions were combined to obtain 15 fractions, which were denoted as Fr.5-1 to Fr.5-15. (6) The fraction Fr.5-15 (161.7 mg) was eluted with methanol on a Sephadex LH-20 gel column. Each tube was collected in 8 mL. The fractions were detected by thin-layer chromatography (TLC). Similar fractions were combined to obtain 5 fractions, which were named Fr.5-15-1 to Fr.5-15-5. (7) Fr.5-15-2 (14.0 mg) was eluted by semi-preparative high performance liquid chromatography (C18; acetonitrile / water, 50:50, V / V; flow rate 4.0 mL / min; UV detection wavelength 210 / 254 nm) to obtain chromone and strophane-type sesquiterpene heteropolymers with molecular weights between 500-700 Da (tR 66.0 min; 2.8 mg).

[0047] The structures of the prepared chromone and scoralaspirane-type sesquiterpene heteropolymers were identified, among which the compound with structure I... 1 H NMR spectrum as follows Figure 1 As shown, the compound with structure I 13 C NMR spectra as follows Figure 2 As shown, the HSQC spectrum of the compound with structure I is as follows: Figure 3 As shown, the compound with structure I 1 H- 1 H COSY spectrum as follows Figure 4 As shown, the HMBC spectrum of the compound with structure I is as follows: Figure 5As shown, the ROESY spectrum of the compound with structure I is as follows: Figure 6 As shown, the HRESIMS spectrum of the compound with structure I is as follows: Figure 7 As shown, the results indicate that the high-resolution mass spectrometry of chromone and strophane-type sesquiterpene hybrids... m / z is 561.2839 [M+Na] + The molecular formula is C 32 H 42 O7; The product structure is shown in Formula I:

[0048] Formula I.

[0049] Chromone and strychnine-type sesquiterpene hybrids 1 H (500 MHz) and 13 The C NMR (125 MHz) data are shown in Table 1: Table 1. Chromone and Acorus spirulane-type sesquiterpene hybrids 1 H and 13 C NMR data ( δ in ppm, J (in Hz, in CD3OD)

[0050] Example 2: Assay of the inhibitory activity of chromone and calamus spirulane-type sesquiterpene hybrids against Helicobacter pylori Frozen Helicobacter pylori strain ( H. pylori (Nanjing Ouke Biotechnology Co., Ltd.) Thaw the bacterial culture at room temperature, inoculate the culture onto a solid agar plate for resuscitation, and incubate in an inverted incubator at 5% O2, 10% CO2, 85% N2, and 37℃ for 5-7 days. Scrape off the cultured colonies and incubate in liquid medium for 4-5 days. Take the cultured Helicobacter pylori strain culture, dilute it with liquid medium (BHI, Haibo Biotechnology HBKP8297-5), and adjust the bacterial concentration to 1×10⁻⁶. 8 CFU / mL. The compound (a chromone and strychnine-type sesquiterpene hybrid prepared in Example 1) was dissolved in dimethyl sulfoxide (DMSO), and the initial screening drug concentration was set at 100 μmol·L⁻¹. −1 At this concentration, the anti-Hp activity of all chromones and strychnine-type sesquiterpene hybrids was tested.

[0051] The antibacterial rate test method used microdilution to remove inhibitory concentrations greater than 100 μmol·L⁻¹. -1 The concentration of the compound was adjusted to 3.125–100 μmol·L using a serial dilution method. -1The final volume was 200 μL / well. Amoxicillin, a commonly used antibiotic in triple or quadruple therapy for Helicobacter pylori, was used as the positive control, while BHI and DMSO served as blank controls. Each experimental group was repeated in triplicate. The 96-well plates were incubated at 37°C with microaerophilic conditions and shaken at 150 rpm for 48 h. The absorbance was measured at 600 nm using a microplate reader, combined with the turbidity in the wells. The MIC was the lowest concentration at which no bacterial growth was observed; the MBC was the lowest concentration at which bacteria in the culture medium could be killed.

[0052] Table 2. Results of the inhibitory activity of chromone and strychnine-type sesquiterpene hybrids against Helicobacter pylori.

[0053] As can be seen from Table 2, although the MIC and MBC of the chromone and sesquiterpene hybrids in this application are slightly higher than those of the positive control group amoxicillin, amoxicillin, as a synthetic antibiotic, has adverse reactions such as allergic reactions and gastrointestinal irritation. Moreover, long-term overuse has led to bacterial resistance and can also disrupt the body's normal flora. The chromone and strychnine-type sesquiterpene hybrids in this invention are the main active ingredients among the two major characteristic components of agarwood, chromone and sesquiterpene. Their separation conditions are relatively mild, resulting in less pollution and a more prominent green attribute compared to chemical synthesis. Furthermore, the single natural compound exhibits better activity. As isolates from natural plants, the chromone and sesquiterpene hybrids in this application have great potential for treating or alleviating gastric diseases caused by Helicobacter pylori infection. In addition, they are safe to obtain, have low toxicity, low irritation, few adverse reactions, and good tolerance to the body. Sri Lankan Wara agarwood can be artificially cultivated and harvested cyclically, providing a stable and sustainable source of raw materials, making it more promising for the development of green antibacterial agents.

[0054] As can be seen from Examples 1 and 2 above, the present invention has discovered a new type of chromone and calamus spirulane-type sesquiterpene hybrid compound with inhibitory activity against Helicobacter pylori; this compound is derived from Sri Lankan Wara agarwood and has good application prospects in the development of drugs for treating chronic gastritis, atrophic gastritis and gastric ulcers caused by Helicobacter pylori infection.

[0055] The above embodiments are only used to illustrate the present invention, and the scope of protection of the present invention is not limited to the above embodiments. Those skilled in the art can achieve the purpose of the present invention based on the above disclosure. Any improvements and modifications made based on the concept of the present invention fall within the scope of protection of the present invention, and the specific scope of protection is determined by the claims.

Claims

1. A chromone and calamus spirulane-type sesquiterpene hybrid compound with inhibitory activity against Helicobacter pylori, characterized in that, The chromone and calamus spirulane type sesquiterpene hybrid has the structure shown in Formula I: ， Formula I; The preparation of the chromone and strychnine-spirulane type sesquiterpene hybrid includes the following steps: (1) After pulverizing 384.0g of Sri Lankan Wara agarwood sample, it was extracted five times by reflux with 2.5L of ethanol aqueous solution with a volume percentage of 95% heated to a slight boiling state. The resulting extracts were filtered and combined, and concentrated under reduced pressure at 45℃ to obtain 129.4g of ethanol extract. (2) Dilute the ethanol extract into a suspension with 1.0L of water, and extract it three times in sequence with petroleum ether, ethyl acetate and n-butanol. The amount of petroleum ether, ethyl acetate and n-butanol used each time is 1.0L. The extract layers are evaporated and concentrated at 45°C to prepare petroleum ether phase extract, ethyl acetate phase extract and n-butanol phase extract, respectively. (3) Take 107.8g of ethyl acetate extract and pass it through a silica gel H reduced pressure normal phase silica gel column with gradient elution of chloroform:methanol volume ratio of 200:1, 100:1, 50:1, 40:1, 30:1, 20:1, 10:1, and 5:

1. Collect 250mL of the sample in one bottle. Use 5.0L of mixed solution for each gradient. Detect each fraction by thin-layer chromatography plate. Combine similar fractions to obtain 11 fractions, which are denoted as Fr.1 ~ Fr.

11. (4) Liquid chromatography-mass spectrometry analysis of the Fr.1 ~ Fr.11 components revealed that Fr.5 contained chromone and sesquiterpene heteropolymers with a molecular weight between 500-700 Da; (5) Separate the fraction Fr.5 3.2g by chromatography on a C18 reversed-phase column of 4.0cm×30.0cm. Elute sequentially with methanol-water with methanol volume percentages of 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%. Collect 100mL in one bottle. Use 1.5L of mixed solution for each gradient. Detect each fraction by thin-layer chromatography plate. Combine similar fractions to obtain 15 fractions, which are recorded as Fr.5-1~Fr.5-15. (6) The fraction Fr.5-15 161.7mg was eluted with methanol on a Sephadex LH-20 gel column. Each tube was collected in 8mL. The fractions were detected by thin-layer chromatography plate. Similar fractions were combined to obtain 5 fractions, which were named Fr. 5-15-1~Fr.5-15-5. (7) 14.0 mg of Fr.5-15-2 was eluted with acetonitrile-water at a volume ratio of 50:50 on a C18 semi-preparative high performance liquid chromatography column at a flow rate of 4.0 mL / min and UV detection wavelengths of 210 and 254 nm. The fraction with a retention time tR of 66.0 min was collected to obtain 2.8 mg of chromone and strophane-type sesquiterpene heteropolymer with a molecular weight between 500-700 Da.

2. A drug that inhibits Helicobacter pylori activity, characterized in that, It includes the chromone and strophanthiolane-type sesquiterpene hybrids as described in claim 1, as well as pharmaceutically acceptable excipients.

3. The use of the chromone and calamus spirulane type sesquiterpene hybrid compound according to claim 1 in the preparation of a medicament for treating and / or alleviating diseases related to Helicobacter pylori.

4. The application according to claim 3, characterized in that, The diseases associated with Helicobacter pylori are selected from chronic gastritis and gastric ulcer.

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