Group of methsadazole compounds as well as preparation method and application thereof

Abstract

The invention discloses a group of methsadazole compounds which are one of compounds with a structure as shown in a formula (I). The compound is obtained by culturing and fermenting Myxococcus sp. SDU36CCTCC NO: M 2021520, and separating from the fermentation liquid. Experiments prove that the methsadazole compounds disclosed by the invention can obviously promote the generation of blood vessels of zebra fish with damaged blood vessels, and also have a certain relieving effect on thrombus. The compounds disclosed by the invention are expected to gain the application value in promoting angiogenesis and antithrombotic activity, a new way is provided for development and application of preparation of a new generation of cardiovascular system medicines, and the compounds have better social benefits, economic values and market prospects in clinical application.

Description

technical field

[0001] The invention relates to a group of mesadazole compounds, as well as the fermentation, separation and preparation of the mesadazole compounds and their application in the preparation of medicines for treating and preventing cardiovascular diseases. It belongs to the technical field of microbial technology and its products and applications. Background technique

[0002] Myxobacteria are a class of Gram-negative bacteria with complex cell social behavior and multicellular development cycle, capable of gliding, forming morphologically specific fruiting bodies, sporulating and preying on other bacteria, with abundant secondary metabolites and diverse biological activities, More and more attention has been paid by scholars at home and abroad (J. F.J.Marcos-Torres,E.García-Bravo,A. J. Pérez, Front. Microbiol. 2016, 7, 781-781.). Myxobacteria are a rich source of hybrid natural products, the most famous of which is a series of NRPS-PKS hybrid epothilones...

Examples

Embodiment 1

[0061] Example 1: Obtaining, Preserving, and Separation and Purification of Mesadazole Compounds (with Mesadazole A 1 , Mesadazole A 2 , Mesadazole B 2 example)

[0062] 1.1 Isolation of strains

[0063] The isolation method is as follows: After the WCX medium has solidified, smeared with E. coli. The fresh or air-dried samples (the soil samples from the lake bottom of Dongcuo Lake, Gaize County, Tibet Autonomous Region) were evenly spread on the E. coli bacteria pads. After culturing to the 3rd day, the plates were checked under a stereo microscope every day for the appearance of fruiting bodies or biofilms of myxobacteria. If there is, carefully pick the top of the fruiting body or scrape the edge of the pellicle, and transfer it to a new WCX medium plate with E. coli pads or VY / 2 medium for purification. In the experiment, a yellow bacterium was obtained, named SDU36.

[0064] Wherein, the formula of above-mentioned WCX medium: CaCl 2 ·2H 2 O 0.1g / 100ml, agar 1.5g / 1...

Embodiment 2

[0086] Example 2 Structure identification of compound 1, compound 2 and compound 3

[0087] 2.1 Identification of the planar structure of compound 1

[0088] High resolution mass spectrometry gives the quasi-molecular ion peak [M+H] of compound 1 + m / z546.3168 (calcd 546.3174), so it is speculated that its molecular weight is 545, and its molecular formula is C 29 H 44 N 3 O 7 , the desaturation is 10.

[0089] Infrared absorption spectrum at 3340cm respectively -1 and 1563cm -1 The characteristic absorption bands of hydroxyl (associative) and strong N–O stretching vibrations are shown at .

[0090] DEPTQ spectrum of compound 1 ( 13 C spectrum) gives 29 carbon signals: including 4 methyl groups, 9 methylene groups, 5 methine groups (three oxygens), 10 aromatic carbons and 1 carboxyl group (Table 1) . 1 H– 1 The H COSY and related HMBC signals indicate that the structure contains three sets of spin-coupling systems, which are: the fatty chain from C-1' to C-7', the f...

Embodiment 3

[0120] Example 3 Test of pro-angiogenic activity of compounds 1-3

[0121] Healthy zebrafish larvae (AB line) and transgenic line Tg (fli1-EGFP) were provided by the Institute of Biology, Shandong Academy of Sciences. According to the Zebrafish Handbook, they were incubated in a closed-flow system of activated carbon-filtered tap water at 28°C with 10-hour dark and 14-hour light intervals. AB line wild-type zebrafish can be used for the observation of thrombus. Because of the expression of green fluorescent protein (GFP) in endothelial cells of transgenic zebrafish Tg (fli1-EGFP), it can be used for the observation of angiogenesis experiments.

[0122] The methods of angiogenic activity refer to the literature (M.Zhang, P.Li, F.Wang, S.Zhang, H.Li, Y.Zhang, X. Wang, K.Liu, X.Li, Food Funct. 2021, 12 , 2282-2291.). The PTK787-induced zebrafish intersegmental vascular (ISV) injury model was used to evaluate the effects of compounds 1–3 on angiogenesis.

[0123]The same paren...

technical field

[0001] The invention relates to a group of mesadazole compounds, as well as the fermentation, separation and preparation of the mesadazole compounds and their application in the preparation of medicines for treating and preventing cardiovascular diseases. It belongs to the technical field of microbial technology and its products and applications. Background technique

[0002] Myxobacteria are a class of Gram-negative bacteria with complex cell social behavior and multicellular development cycle, capable of gliding, forming morphologically specific fruiting bodies, sporulating and preying on other bacteria, with abundant secondary metabolites and diverse biological activities, More and more attention has been paid by scholars at home and abroad (J. F.J.Marcos-Torres,E.García-Bravo,A. J. Pérez, Front. Microbiol. 2016, 7, 781-781.). Myxobacteria are a rich source of hybrid natural products, the most famous of which is a series of NRPS-PKS hybrid epothilones...

Examples

Embodiment 1

[0061] Example 1: Obtaining, Preserving, and Separation and Purification of Mesadazole Compounds (with Mesadazole A 1 , Mesadazole A 2 , Mesadazole B 2 example)

[0062] 1.1 Isolation of strains

[0063] The isolation method is as follows: After the WCX medium has solidified, smeared with E. coli. The fresh or air-dried samples (the soil samples from the lake bottom of Dongcuo Lake, Gaize County, Tibet Autonomous Region) were evenly spread on the E. coli bacteria pads. After culturing to the 3rd day, the plates were checked under a stereo microscope every day for the appearance of fruiting bodies or biofilms of myxobacteria. If there is, carefully pick the top of the fruiting body or scrape the edge of the pellicle, and transfer it to a new WCX medium plate with E. coli pads or VY / 2 medium for purification. In the experiment, a yellow bacterium was obtained, named SDU36.

[0064] Wherein, the formula of above-mentioned WCX medium: CaCl 2 ·2H 2 O 0.1g / 100ml, agar 1.5g / 1...

Embodiment 2

[0086] Example 2 Structure identification of compound 1, compound 2 and compound 3

[0087] 2.1 Identification of the planar structure of compound 1

[0088] High resolution mass spectrometry gives the quasi-molecular ion peak [M+H] of compound 1 + m / z546.3168 (calcd 546.3174), so it is speculated that its molecular weight is 545, and its molecular formula is C 29 H 44 N 3 O 7 , the desaturation is 10.

[0089] Infrared absorption spectrum at 3340cm respectively -1 and 1563cm -1 The characteristic absorption bands of hydroxyl (associative) and strong N–O stretching vibrations are shown at .

[0090] DEPTQ spectrum of compound 1 ( 13 C spectrum) gives 29 carbon signals: including 4 methyl groups, 9 methylene groups, 5 methine groups (three oxygens), 10 aromatic carbons and 1 carboxyl group (Table 1) . 1 H– 1 The H COSY and related HMBC signals indicate that the structure contains three sets of spin-coupling systems, which are: the fatty chain from C-1' to C-7', the f...

Embodiment 3

[0120] Example 3 Test of pro-angiogenic activity of compounds 1-3

[0121] Healthy zebrafish larvae (AB line) and transgenic line Tg (fli1-EGFP) were provided by the Institute of Biology, Shandong Academy of Sciences. According to the Zebrafish Handbook, they were incubated in a closed-flow system of activated carbon-filtered tap water at 28°C with 10-hour dark and 14-hour light intervals. AB line wild-type zebrafish can be used for the observation of thrombus. Because of the expression of green fluorescent protein (GFP) in endothelial cells of transgenic zebrafish Tg (fli1-EGFP), it can be used for the observation of angiogenesis experiments.

[0122] The methods of angiogenic activity refer to the literature (M.Zhang, P.Li, F.Wang, S.Zhang, H.Li, Y.Zhang, X. Wang, K.Liu, X.Li, Food Funct. 2021, 12 , 2282-2291.). The PTK787-induced zebrafish intersegmental vascular (ISV) injury model was used to evaluate the effects of compounds 1–3 on angiogenesis.

[0123]The same paren...

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