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CN-120081817-B - Chloro-spiro gentinol derivative and preparation method and application thereof

CN120081817BCN 120081817 BCN120081817 BCN 120081817BCN-120081817-B

Abstract

This application discloses a class of chlorinated spirocyclogentian derivatives, their preparation methods, and applications, which belong to the field of analysis technology for marine fungal active ingredients. The chlorinated spirocyclic gentian derivative is a derivative with a hybrid skeleton of chlorogentisyl alcohol and gabosine. The present invention utilizes the polarity difference between chlorogentisyl alcohol and gabosine derivatives to isolate a novel structure of chlorinated spirocyclic gentian derivative from the fermentation culture of marine fungus Epicoccus sorghinum-GST-5. In vitro anti β - glucuronidase assays have shown that the chlorinated spirocyclic gentian derivative provided by the present invention has good β - glucuronidase inhibitory activity and low cytotoxicity, and has development prospects in the preparation of drugs against β - glucuronidase.

Inventors

  • WU BIN
  • GE YICHAO

Assignees

  • 浙江大学

Dates

Publication Date
20260508
Application Date
20220810

Claims (9)

  1. 1. A chlorospiro-gentinol derivative is characterized in that the chlorospiro-gentinol derivative is a derivative with chlorogentisyl alcohol and gabosine heterozygous frameworks, the structural formula is shown in any one of formulas (9), (16) and (17), (9)、 (16)、 (17)。
  2. 2. A process for the preparation of chlorospiro-gentinol derivatives according to claim 1, comprising the steps of: (1) Activating Fucus fungus (Epicoccum sorghinum) GST-5 with a preservation number of CGMCC 3.20238, inoculating the activated Fucus fungus GST-5 into a liquid culture medium, and fermenting and culturing; (2) Separating to obtain mycelium and fermentation liquor after fermentation culture is finished, adding organic solvent into mycelium to extract, separating to obtain extract, concentrating the extract, suspending with distilled water to obtain water suspension, extracting the water suspension with ethyl acetate or n-butanol to obtain extract A; (3) Concentrating the extract A or the extract B, then carrying out normal phase silica gel column chromatography separation, carrying out gradient elution on dichloromethane/methanol mixed liquid with the volume ratio of 100:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1 and 10:1 in sequence, collecting fractions eluted by the dichloromethane/methanol mixed liquid with the volume ratio of 10:1, and carrying out reverse phase silica gel column chromatography and high performance liquid chromatography separation to obtain the derivative.
  3. 3. The method according to claim 2, wherein in the step (1), the liquid medium comprises, based on 1L by volume, 1-5g of starch, 10-20g of bran, 3-15g of yeast extract, 0.1-0.8 of KH 2 PO 4 1-8 g,MgSO 4 •7H 2 O g, and the balance of water; Or the liquid culture medium comprises the following raw materials, by volume of 1L, 200-600g of potatoes, 2-10g of peptone, 1-5 g of yeast extract, 5-20 of glucose g and the balance of water, wherein the initial pH value of the culture solution is 6.0-7.0; Or the liquid culture medium comprises the following raw materials, by volume of 1L, 10-40 g parts of sucrose, 5-20 parts of corn meal g parts of NaNO 3 -4 g parts of NaNO 3 , and the balance of yeast extract 1-4 g、KH 2 PO 4 0.2-0.8 g、MgSO 4 •7H 2 O 0.2-1 g、KCl 0.2-1 g、FeSO 4 0.001-0.005 g, parts of water; Or the liquid culture medium comprises the following raw materials, by volume of 1L, 20-30 g parts of malt extract powder, 15-20 parts of glucose g parts of casein peptone 1-2 parts of g parts of casein peptone and the balance of water.
  4. 4. The method according to claim 2, wherein in the step (1), the condition of the fermentation culture is 20 to 30℃for 10 to 40 days.
  5. 5. The preparation method of claim 2, wherein in the step (3), the fractions are subjected to reverse phase silica gel column chromatography, gradient elution is sequentially carried out on methanol/water mixed solution with volume ratio of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2 and 9:1, each gradient elution is carried out 5 times, serial numbers are sequentially 1-45, fractions 18-25 are combined into a subfraction 7, fractions 26-28 are combined into a subfraction 8, then high performance liquid chromatography separation is carried out, the subfraction 7 is eluted under the methanol/water mixed solution with volume ratio of 42%, the retention time is respectively 18.2 minutes, 24.4 minutes, 26.9 minutes, and 22.3 minutes, the peaks with the structural formulas of 9,11,12 and 10 are respectively shown in formula (9), the peak with the subfraction 8 with volume ratio of 18% acetonitrile/water mixed solution with the retention time is respectively 23.3 minutes, the peak with the retention time of 26.3 minutes is respectively 16 and 17, and the structural formula of 16 is respectively shown in formula (17).
  6. 6. Use of a chlorospirocyclic gentian derivative according to claim 1 in the manufacture of a medicament against β -glucuronidase.
  7. 7. The use according to claim 6, wherein the beta-glucuronidase is enterobacteria beta-glucuronidase.
  8. 8. The use of a chlorospiro gentinol derivative according to claim 1 in the manufacture of a medicament for alleviating gastrointestinal side effects caused by irinotecan, indomethacin medicaments.
  9. 9. The application of the compound with the structural formula shown in the formula (12) in preparing the anti-tumor medicament is characterized in that the tumor is lymphoma, (12)。

Description

Chloro-spiro gentinol derivative and preparation method and application thereof The application discloses a polyhydroxy cyclohexane derivative with application number 202210955910.X, application day 2022, 8 and 10, and the application name of the polyhydroxy cyclohexane derivative, a preparation method and application of the polyhydroxy cyclohexane derivative. Technical Field The invention relates to the technical field of marine fungus active ingredient analysis, in particular to a derivative with gabosine and chlorogentisyl alcohol heterozygous skeletons extracted from epiphyte (Epicoccum sorghinum) GST-5 and application thereof. Background Marine microorganisms are able to withstand extreme conditions such as high salt, high pressure, low oxygen, low light, etc., which are characteristic of the ocean, relative to land microorganisms, and the specificity of living environments leads to diversity of marine microorganisms in species, genetic composition, and ecological functions. The special nature of the marine environment and the progress of the marine microorganism resource acquisition technology bring unprecedented opportunities to the research of natural medicine source compounds of marine microorganism sources. Marine fungi in marine microorganisms are a rich source of active secondary metabolites, 70-80% of which have biological activity, including small-molecule lactone compounds, mycotoxins, novel substances with inhibitory activity on the central nervous system, 1-dodecanol, unsaturated hydrocarbons, acids and esters, and lipopeptid antibiotics capable of inhibiting plant and human mycoviruses from acting on fungal cell walls to synthesize novel targets. The natural product with specific structure type is found to have important significance for research and development of marine drugs by taking marine fungi as raw materials. Beta-Glucuronidase (GUS) is an important class of hydrolytic enzymes produced by the intestinal flora, which catalyze the hydrolysis of glucuronic acid conjugates and the production of the corresponding aglycones. In the human body, GUS distributed in the intestinal tract can also catalyze the hydrolysis of various medicines (such as indomethacin, diclofenac and active products of irinotecan, namely 7-ethyl-10-hydroxycamptothecin) and glucuronide of endogenous hormone, and released aglycone can be absorbed by the intestinal tract again, so that a pharmacokinetics bimodal phenomenon is formed. Notably, this process may cause serious gastrointestinal adverse effects, for example, GUS may catalyze the hydrolysis of 7-ethyl-10-hydroxycamptothecin-O-glucuronic acid (SN-38G) and produce SN-38, a product with potent cytotoxic effects, which accumulates in the intestinal tract and may cause intestinal mucosa to fall off and cause serious adverse effects such as delayed diarrhea. Therefore, the development of high-efficiency and safe GUS inhibitors is of great importance for relieving the lethal diarrhea caused by medicines such as irinotecan and indomethacin (Weng Zaimiao, lv Shanshan, ge Anbo, wang Ping, hou Jie, pharmaceutical progress, research progress of intestinal bacteria beta-glucuronidase inhibitors, 2018,42 (03)). There is currently little research on the chemical composition of epiphyte fungi (Epicoccum). Hao Baocong and the like are separated from dandelion endophytic fungi Epicoccum sorghinum to obtain (4R, 5R, 6S) -4, 5-dihydroxyl-6- (6 '-methylsalicyloxy) -2-methoxymethyl-2-cyclohexene-1-one, (4R, 5R, 6S) -4, 5-dihydroxyl-6- (6' -methylsalicyloxy) -2-hydroxymethyl-2-cyclohexene-1-one, (-) -gabosine E, theobroxide, 3-chlorogentinol and 3-hydroxybenzyl alcohol (Hao Baocong, zheng Yaoyao, chen Xu, chen Qiuxia, ji Renan, chen Min, pharmaceutical science report, and the dandelion endophytic fungi Epicoccum sorghinum-2 were subjected to secondary metabolite research, 2022,57 (07)). The meta-superclass researches on chemical components of a strain of endophytic fungus Epicoccus nigrum Epicoccum nigrum one separated from lichen Leptogium masiaticum, respectively identified as 1 alkaloid compound fusaricide,7 benzofuran compounds epicoccone B,4,6-dihydroxy-5-methoxy-7-methyl-1,3-dihydro isobenzofuran,5-methyl- epicoccone B,3,6,7-trihydroxy-5-methoxy-4- methylisobenzo furan-1(3H)- one ,3-methoxyepicoccone B,2,3,4-trihydroxy -6-(hydroxymethyl)-5- methylbenzyl-alcohol ,isoochracinic acid and 3 epicoccolide compounds epicocconigrones A, epicoccolide B and epicocconigrones B, wherein alkaloids fusaricide and epicoccolide compounds have strong antibacterial activity, and alkaloid fusaricide has better in-vitro inhibitory activity on lung cancer cells M109 (meta-superclass Guo Yuhua, zhang Yingbo, hu Xuan and Wang Dan, fulai, li Gang, chinese journal of traditional Chinese medicine, research on secondary metabolites of a strain of plant endophytic fungus Epicoccum nigrum one, 2019,44 (18), 4021-4025). However, the isolation of chlorogentisyl alcohol and gabosine anti-GUS enzyme compou