阅读说明：本技术 一种海藻倍半萜类化合物在制备抑菌药物中的用途 (Application of seaweed sesquiterpene compound in preparation of antibacterial drugs ) 是由 贾睿 苏永政 钱恒 何培民 刘萌萌 蔡春尔 于 2019-10-17 设计创作，主要内容包括：本发明公开了一种海藻倍半萜类化合物在制备抑菌药物中的用途,该化合物来源于海洋红藻冈村凹顶藻(Laurencia okamurai),其分子式为C<Sub>15</Sub>H<Sub>19</Sub>O<Sub>2</Sub>Br,结构式如下式所示：<Image he="320" wi="445" file="DDA0002237185910000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>本发明首次发现海藻倍半萜类化合物对副溶血性弧菌和溶藻弧菌显示出很好的抑制活性,与广谱抗生素相比,其作为新型抑菌剂对副溶血性弧菌和溶藻弧菌具有特异性,对其他微生物影响小,不影响微生态,虽然抗菌效果比抗生素效果稍弱,但其低毒、环境友好、纯天然和不产生耐药性,应用前景广阔。(The invention discloses an application of a seaweed sesquiterpene compound in preparation of antibacterial drugs, wherein the compound is derived from Laurencia okamurai (Laurencia okamurai), and the molecular formula of the compound is C 15 H 19 O 2 Br, formula shown below: the invention discovers for the first time that the alga sesquiterpenoids have good inhibitory activity on vibrio parahaemolyticus and vibrio alginolyticus, compared with broad-spectrum antibiotics, the alga sesquiterpenoids have specificity on vibrio parahaemolyticus and vibrio alginolyticus, have small influence on other microorganisms, do not influence on microecology, have low toxicity, environmental friendliness, pure nature and no influence although the antibacterial effect is slightly weaker than that of antibioticsDrug resistance is generated, and the application prospect is wide.)

1. The application of the seaweed sesquiterpene compound in preparing the antibacterial drug is characterized in that the seaweed sesquiterpene compound is derived from sea red algae Laurencia okamurai (Laurencia okamurai), and the molecular formula of the seaweed sesquiterpene compound is C₁₅H₁₉O₂Br, formula (I):

2. the use of claim 1, wherein the bacteria are vibrio parahaemolyticus and vibrio alginolyticus, and the inhibition of the vibrio parahaemolyticus and vibrio alginolyticus activity by the algal sesquiterpene compound is used.

3. Use according to claim 1 or 2, characterized in that said seaweed sesquiterpenoids are prepared by a process comprising:

(1) drying and crushing Laurencia okamurana in Okamura village in shade, extracting with an organic solvent A for 2-5 times, combining the extracting solutions, concentrating, extracting with ethyl acetate, and concentrating to obtain a crude extract; wherein:

the organic solvent A is one or more of dichloromethane (AR), Acetone (AR), chloroform (AR), ethyl Acetate (AR), ethanol (AR) or methanol (AR);

(2) carrying out silica gel column chromatography on the crude extract in the step (1), carrying out gradient elution by using an organic solvent B, and collecting eluent; wherein:

the organic solvent B is a mixture (AR) of petroleum ether and acetone in any volume ratio, the gradient elution is carried out according to the volume ratios of the petroleum ether to the acetone of 100:0, 99:1,98:2,95:5,9:1,8:2 and 7:3 respectively, and the elution part with the volume ratio of the petroleum ether to the acetone of 9:1 is collected;

(3) separating and purifying the collected eluate by gel column and thin layer chromatography, and collecting R_fObtaining a component with a value of 0.5-0.6 to obtain the seaweed sesquiterpenoids shown in the formula (I); wherein:

performing TLC detection by using chloroform-methanol mixed solvent with the volume ratio of 1:1 as chromatographic eluent of the gel column, developing the chromatographic eluent by using vanillin-sulfuric acid, and collecting red spot components;

the developing agent for the thin layer chromatography is a petroleum ether-acetone mixture with the volume ratio of 9: 1.

Technical Field

The invention relates to the field of biological medicine, in particular to application of seaweed sesquiterpene compounds with a structural formula shown in (I) in preparation of bacteriostatic agents for inhibiting vibrio parahaemolyticus and vibrio alginolyticus.

Background

The Graptospira okamurai (Laurencia okamurai) belongs to Rhodophyta, Rhodophyceae, Ceramiales, Arthropodaceae, and Laurencia in classification status, is mainly distributed in coastal areas of Pacific ocean such as China, Japan, and Korean peninsula, and generally grows on rocks in intertidal zones (Shidao, New Zea, Xifeng, etc. oceanic science, 2007,31(4):81-91.Masuda M.; AbeT.; Suzuki T.Phytology, 1996,35(6): 550-562.). The sesquiterpene compound is the most abundant compound in secondary metabolites of Laurencia gambogonii, and has various biological activities.

Vibrio parahaemolyticus (Vibrio parahaemolyticus) belongs to Vibrio of Vibrionaceae, is gram-negative bacillus, is a food-borne pathogenic bacterium, is widely existed in sea water, submarine sediments, marine products such as fish, shellfish, shrimp, crab and the like, and people eating seafood containing the pathogenic bacterium can cause food poisoning, cause symptoms such as abdominal pain, vomiting, diarrhea and the like, and belongs to zoonosis pathogenic bacteria. Vibrio alginolyticus (Vibrio alginilyticus) is halophilic gram-negative short bacillus, is a common marine pathogenic bacterium, has biological characteristics similar to that of Vibrio parahaemolyticus, can cause diseases such as wound infection, food poisoning, tympanitis and the like to people, is also a conditional pathogenic bacterium of marine cultured animals such as fish, shrimp, shellfish and the like, and is easy to outbreak when the organism immune function is reduced when the environment is deteriorated.

At present, antibiotics are mainly used for treating the two germs, but the antibiotics easily cause drug resistance of the germs, and the long-term use of the antibiotics can destroy the ecological environment. The alga sesquiterpene compound as a bacteriostatic agent has the advantages of low toxicity, pure nature and the like, researches show good ecological benefit, economic benefit and social benefit, and the application of the alga sesquiterpene compound has great influence on the aquaculture industry and the human medical industry.

Disclosure of Invention

The invention discovers that the seaweed sesquiterpenoid compound aplysinol shows good inhibitory activity to vibrio parahaemolyticus and vibrio alginolyticus for the first time, and provides the application of the natural seaweed sesquiterpenoid compound in preparing antibacterial drugs, in particular to the preparation of antibacterial agents for inhibiting the activity of vibrio parahaemolyticus and vibrio alginolyticus.

The sesquiterpenoids are derived from Laurencia okamurai (Laurencia okamurai) and have molecular formula of C₁₅H₁₉O₂Br, formula (I):

the preparation method of the seaweed sesquiterpenoids comprises the following steps:

(1) drying and crushing Laurencia okamurana in Okamura village in shade, extracting with an organic solvent A for 2-5 times, combining the extracting solutions, concentrating, extracting with ethyl acetate, and concentrating to obtain a crude extract; wherein the organic solvent A is one or more of dichloromethane (AR), Acetone (AR), chloroform (AR), ethyl Acetate (AR), ethanol (AR) or methanol (AR);

(2) carrying out silica gel column chromatography on the crude extract in the step (1), carrying out gradient elution by using an organic solvent B, and collecting eluent; wherein the organic solvent B is a mixture (AR) of petroleum ether and acetone in any volume ratio, the gradient elution is carried out according to the volume ratios of the petroleum ether to the acetone of 100:0, 99:1,98:2,95:5,9:1,8:2 and 7:3 respectively, and the elution part with the volume ratio of the petroleum ether to the acetone of 9:1 is collected;

(3) separating and purifying the collected eluate by gel column and thin layer chromatography, and collecting R_fObtaining a component with a value of 0.5-0.6 to obtain the seaweed sesquiterpenoids shown in the formula (I); wherein, the chromatographic eluent of the gel column is a chloroform-methanol mixed solvent with the volume ratio of 1:1, TLC detection is carried out, vanillin-sulfuric acid is used for color development, and red spot components are collected; the developing agent for the thin layer chromatography is a petroleum ether-acetone mixture with the volume ratio of 9: 1.

Compared with the prior art, the invention has the following beneficial effects:

1. the invention discovers for the first time that the seaweed sesquiterpenoid compound aplysinol shows good inhibitory activity to vibrio parahaemolyticus and vibrio alginolyticus, and the compound belongs to a small molecular natural organic compound, is easy to prepare and obtain and can be used as a novel bacteriostatic agent.

2. Compared with broad-spectrum antibiotics, the alga sesquiterpenoid compound aplysinol has specificity on vibrio parahaemolyticus and vibrio alginolyticus, has small influence on other microorganisms, and does not influence the microecology. Although the antibacterial effect is slightly weaker than that of antibiotics, the seaweed sesquiterpenoids are low in toxicity, environment-friendly, pure natural and free of drug resistance, and have wide prospects when used for preparing novel antibacterial drugs in the form that the current domestic antibiotic drugs are required to be used carefully in aquaculture industry and medical industry.

Drawings

FIG. 1 shows the preparation of algal sesquiterpene compound aplysinol¹HNMR spectrogram.

FIG. 2 shows the bacteriostatic effect of the algal sesquiterpene compound aplysinol (Vibrio parahaemolyticus, Vibrio alginolyticus).

Detailed Description

In order to make the technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be described in detail below.