A catechol compound and its preparation method and application

Abstract

The present invention relates to the fields of microbiology and pharmaceutical technology, and in particular to a catechol compound, a preparation method, and applications thereof. A catechol compound characterized by having the following structural formula: wherein R1 represents a methyl group, an ethyl group, an isopropyl group, a benzyl group, or a p-hydroxybenzyl group; and R2 represents a benzyl group. The present invention provides a catechol compound, a preparation method, and applications thereof. The catechol compound is a natural product obtained by microbial fermentation, has excellent anti-fibrosis and anti-inflammatory activities, and has promising clinical application prospects for anti-fibrosis and anti-inflammatory treatments.

Description

Technical Field

[0001] The present invention relates to the technical fields of microbiology and pharmacy, and in particular to a catechol compound and a preparation method and application thereof. Background Art

[0002] Catechol is the common name for catechol. Microbial-derived catechol-type iron carriers are usually catecholamide compounds formed by the condensation of one or more 2,3-dihydroxybenzoic acid (DHBA) with amino acids or short peptides and dehydration. For example, enterobactin orenterochelin found in Escherichia coli contains 3 DHBs and 6 phenolic hydroxyl groups in its structure, which can react with trivalent iron ions (Fe 3+) form hexadentate octahedral ion chelates, or iron-siderophore complexes, that assist microorganisms in transporting iron ions into cells and exerting their biological functions. Catechol-type siderophore compounds are often named after the producing bacteria, such as Serratiochelins, which are produced by Serratia strains. Myxochelins are a class of catechol-type siderophore compounds primarily produced by myxobacteria. They include myxochelin AD, hyalachelins AC, myxochelin N and O, myxochelin N1, and myxochelin B succinate. In addition to the 11 myxochelins produced by myxobacteria mentioned above, a new myxochelin, Pseudochelin A, was discovered from Pseudoalteromonas piscicida S2040. The synthesis of natural myxochelins relies on the catalysis of nonribosomal peptide synthetases (NRPSs). With the assistance of enzymes MxcE and MxcF, the nonribosomal peptide synthetase (NRPS) MxcG links two 2,3-DHBA units to the amino group of L-lysine. The resulting thioester intermediate is reduced to an aldehyde intermediate by the reductase domain of mxcG, which is further reduced to form myxochelin A or catalyzed by an aminotransferase (MxcL) to form myxochelin B. 2,3-DHBA is catalyzed by the protein encoded by the biosynthetic gene mxcCDEF. Studies have shown that the key synthase MxcE in some strains is not specific for the substrate 2,3-DHBA, but rather exhibits a certain tolerance. This results in the formation of atypical myxochelins such as Myxochelin N and Myxochelin O. Although these lack iron-chelating groups and are not siderophores, their biosynthetic pathways are consistent with those of typical myxochelins and are still referred to as myxochelins. Targeting this type of substrate-tolerant strain, a precursor-directed biosynthesis (PDB) strategy was adopted. By adding structural analogs of the substrate 2,3-DHBA, 31 non-natural mycochelin compounds have been obtained.

[0003] Previous studies have shown that mucochelins possess antimicrobial, antioxidant, and antitumor activities. Mucochelins have been most extensively studied as inhibitors of 5-lipoxygenase (5-LOX). As a key upstream enzyme in the production of leukotrienes (LT), 5-LOX has become an important target for drugs treating these diseases. A-64077, also known as Zileuton, developed by Abbott Pharmaceuticals in the United States, is currently the only marketed 5-LOX inhibitor, primarily for the prevention and treatment of chronic asthma in patients aged 12 years and older. Furthermore, research on mucochelin activities against tumor cell proliferation, metastasis, and invasion is also a focus. Studies have shown that myxochelin A, at non-cytotoxic concentrations, significantly inhibits the metastasis and invasion of mouse colon cancer 26-L5 cells. Summary of the Invention

[0004] The present invention provides a catechol compound and a preparation method and application thereof, which are used for anti-fibrosis and anti-inflammatory treatments.

[0005] In a first aspect, the present invention first provides a catechol compound having any of the following structural formulas:

[0006]

[0007] Wherein, R1 represents methyl, ethyl, isopropyl, benzyl or p-hydroxybenzyl; R2 represents benzyl.

[0008] The present invention first discovered mycochelins with modified hydroxyl groups on their aliphatic chains. Since they were first discovered from secondary metabolites of Micromonospora, they were named sporachelins. Studies on the activity of sporachelins have shown for the first time that they have a strong and concentration-dependent inhibitory effect on the expression of fibrosis marker genes COL1A1, ACTA2, and TGFB1 in human hepatic stellate cells (LX-2), and have low cytotoxicity against human hepatic stellate cells (CC). 50 >200 μM), suggesting its potential use in anti-fibrosis therapy. Furthermore, at low concentrations, sporechelin also exhibited a strong inhibitory effect on human 5-lipoxygenase activity and the expression of inflammatory marker genes Tnfα and Cox2 in mouse peritoneal macrophages Raw264.7, suggesting its potential use in anti-inflammatory therapy.

[0009] As a preferred embodiment of the present invention, the catechol compound comprises:

[0010] The molecular formula of sporechelin A is C 28 H 30 N2O8, molecular weight is 522; the molecular formula of spore chelatein A1 is C27 H 29 N3O6, molecular weight is 491; the molecular formula of spore chelate is C 28 H 30 N2O9, molecular weight is 538; the molecular formula of spore chelate is C 22 H 26 N2O8, molecular weight is 446; the molecular formula of spore chelate is C 23 H 28 N2O8, molecular weight is 460; the molecular formula of spore chelate is C 24 H 30 N2O8, molecular weight is 474.

[0011] In a second aspect, the present invention provides the use of the catechol compounds in the preparation of anti-fibrosis drugs.

[0012] In the present invention, pharmaceutically acceptable salts of catechol compounds also fall within the scope of protection of the present invention. The catechol compounds of the present invention can be used in the form of pharmaceutically acceptable salts derived from inorganic acids or organic acids. The term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., within the scope of reliable medical judgment, and are commensurate with a reasonable effect / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge, et al., J. Pharmaceutical Sciences, 1977, 66: 1 provide a detailed description of pharmaceutically acceptable salts. The salts can be prepared by reacting the free base functionality of the compound of the present invention with a suitable organic acid.

[0013] In a third aspect, the present invention further provides use of the pharmaceutically acceptable salt of the catechol compound in the preparation of anti-fibrosis drugs.

[0014] In a fourth aspect, the present invention further provides use of the catechol compounds or pharmaceutically acceptable salts thereof in the preparation of anti-inflammatory drugs.

[0015] In a fifth aspect, the present invention further provides an anti-fibrosis drug containing the catechol compound or a pharmaceutically acceptable salt thereof.

[0016] In a sixth aspect, the present invention further provides an anti-inflammatory drug containing the catechol compound or a pharmaceutically acceptable salt thereof.

[0017] In the present invention, the anti-fibrosis drug or anti-inflammatory drug can be a drug for treating and / or preventing the disease.

[0018] In the present invention, the anti-fibrosis or anti-inflammatory drug may contain, in addition to the catechol compound or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier may account for 0.1 to 99.9% by weight of the total weight of the anti-fibrosis or anti-inflammatory drug.

[0019] In the present invention, the pharmaceutically acceptable carrier includes one or more of microcrystalline cellulose, sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, mannitol, sorbitol, sorbic acid or potassium salt, surfactant, polyethylene glycol, cyclodextrin, β-cyclodextrin, phospholipid material, kaolin, talc, calcium stearate, magnesium stearate and microcrystals.

[0020] The anti-fibrosis drug or anti-inflammatory drug of the present invention can be prepared into any desired dosage form, such as tablets, capsules, injections, ointments, creams, and sprays.

[0021] In the present invention, the active ingredient of the anti-fibrosis drug or anti-inflammatory drug includes the catechol compound or a pharmaceutically acceptable salt thereof. The active ingredient of the above-mentioned anti-liver fibrosis drug or anti-inflammatory drug may also contain other biological components or non-biological components. Those skilled in the art can determine the other active ingredients based on the anti-fibrosis or anti-inflammatory effects.

[0022] In a seventh aspect, the present invention further provides the use of the catechol compounds in treating fibrotic diseases or inflammation.

[0023] In the present invention, the fibrotic disease may be caused by inflammation, and may include liver fibrosis, lung fibrosis, kidney fibrosis or cardiac fibrosis.

[0024] The compounds sporechelin AE and sporechelin A1 of the present invention have a good and concentration-dependent inhibitory effect on the expression of fibrosis marker genes COL1A1, ACTA2 and TGFB1 in human hepatic stellate cells LX-2 at concentrations of 1 μM and 5 μM, and have low cytotoxicity to human hepatic stellate cells LX-2 (CC 50 >200μM), and can be used for the treatment of liver fibrosis. Furthermore, sporechelin AE and sporechelin A1 both inhibited human 5-lipoxygenase by >50% at a 2μM concentration. Furthermore, at a 1μM concentration, they also significantly inhibited the expression of inflammatory marker genes Tnfα and Cox2 in mouse peritoneal macrophages Raw264.7, making them useful for anti-inflammatory treatments.

[0025] Sporochelin AE and sporochelin A1 in the present invention are natural products obtained through microbial fermentation. The raw materials are readily available, and therefore they can be used as lead compounds for anti-fibrosis or anti-inflammatory treatments, undergoing structural optimization to develop more active anti-fibrosis or anti-inflammatory drugs. Sporochelin AE and sporochelin A1, as active ingredients, can be combined with one or more pharmaceutically acceptable carriers, excipients, or excipients to produce anti-fibrosis or anti-inflammatory pharmaceutical compositions. Sporochelin AE and sporochelin A1, and their pharmaceutical compositions, can be used in the clinical treatment of fibrosis or inflammation.

[0026] In an eighth aspect, the present invention also provides a method for preparing the catechol compounds, comprising S1 to S2, wherein S1: fermenting and culturing Micromonospora to obtain a fermentation broth; S2: extracting and separating the catechol compounds from the fermentation broth, wherein the Micromonospora is Micromonospora sp.TMD166, and its registration number in the General Microorganism Center of the China Culture Collection Administration is CGMCC No. 21622.

[0027] In the present invention, in the preparation method of the catechol compounds, the culture medium can be a culture medium capable of culturing actinomycetes, which can be a solid culture medium, a semi-solid culture medium or a liquid culture medium. The actinomycete culture medium can be an International Streptomyces Program culture medium (ISP2, ISP3, ISP4, etc.), Gao's medium No. 1, trypticase soy broth medium (TSB) or other actinomycete fermentation medium well known to professionals in the field. The ISP2 culture medium can be made of glucose, malt extract, yeast extract and water, or can be made of glucose, malt extract, yeast extract, inorganic salts and water, or can be made of glucose, malt extract, yeast extract, inorganic salts, vitamin complexes and water. The Gao's medium No. 1 can be made of starch, inorganic salts and water, or can be made of starch, inorganic salts, vitamin complexes and water. The trypticase soy broth medium can be made of trypticase peptone, soy peptone, glucose and water, or can be made of trypticase peptone, soy peptone, glucose, inorganic salts and water, or can be made of trypticase peptone, soy peptone, glucose, inorganic salts, vitamin complex and water. The other actinomycete fermentation media well known to professionals in the field can be made of one or more fast-acting or slow-acting carbon sources, one or more fast-acting or slow-acting nitrogen sources, water and / or inorganic salts. Carbon source is a type of nutrient for microbial growth, which is a carbon-containing compound, including fast-acting or slow-acting carbon sources such as sugars, oils, organic acids and organic acid esters and small molecule alcohols. Nitrogen source refers to a substance that provides nitrogen elements required for microbial nutrition, including fast-acting or slow-acting nitrogen sources such as peanut cake powder, soybean cake powder, yeast powder, peptone, ammonia water, ammonium salts and nitrates.

[0028] The preparation method of the catechol compounds can be liquid fermentation preparation.

[0029] In the above-mentioned liquid fermentation for preparing the catechol compounds, the culture medium may be a liquid culture medium, and the fermentation product may be a liquid fermentation product.

[0030] Preferably, the fermentation culture comprises: inoculating the Micromonospora on a seed culture medium for culture, and then transferring the culture medium to a fermentation culture medium for shaking culture.

[0031] Preferably, the culture conditions on the seed culture medium are 28-30°C, 150-200 r / min, and the culture time is 3-4 days; the culture conditions on the fermentation culture medium are 28-30°C, 150-200 r / min, and the culture time is 6-7 days.

[0032] Preferably, the formula of the seed culture medium and the fermentation medium is the same; further preferably, medium No. 38 is selected; further preferably, the formula is: glucose 4.0g, yeast powder 4.0g, malt extract powder 5.0g, ZnSO4.7H2O 0.001g, MnCl2.4H2O 0.001g, FeSO4.7H2O0.002g, phenylalanine 0.001g, alanine 0.0003g, vitamin B1 0.001g, vitamin B2 0.001g, vitamin B6 0.001g, niacin 0.001g, biotin 0.001g, and sterile distilled water 1.0L; the pH of the seed culture medium and the fermentation medium is 7.2.

[0033] Preferably, the S2 includes S21 to S22, S21: centrifuging the fermentation broth to obtain a supernatant and mycelium, taking the supernatant for extraction treatment to obtain a supernatant extract; S22: preliminarily separating the supernatant extract by gel column chromatography, then separating it by reverse-phase silica gel column chromatography, gradient eluting with a solvent, and finally preparing and separating the catechol compounds by HPLC.

[0034] Preferably, during the extraction treatment, the extraction solvent is one or more of chloroform, dichloromethane and ethyl acetate; in one embodiment of the present invention, it is ethyl acetate.

[0035] Preferably, the medium used in the gel column chromatography separation is Sephadex LH-20 or Tosoh Toyopearl HW-40; in one embodiment of the present invention, it is Sephadex LH-20.

[0036] Preferably, the medium used in the reverse phase silica gel column is C18 reverse phase silica gel or phenyl reverse phase silica gel; in one embodiment of the present invention, it is C18 reverse phase silica gel.

[0037] Preferably, the chromatographic column used in the HPLC preparative separation is a C18 chromatographic column, a C8 chromatographic column, a C3 chromatographic column or a phenyl bonded column. In one embodiment of the present invention, it is a C18 chromatographic column or a C8 chromatographic column.

[0038] Preferably, the mobile phase used in the gel column chromatography is one or more of chloroform, dichloromethane, ethyl acetate, alcohols with a carbon chain ≤ 4, acetone and butanone; or a mixed solution of one or more of the above solvents and water; in one embodiment of the present invention, it is methanol.

[0039] Preferably, the solution used for the reverse phase silica gel column chromatography separation is one or more of acetone aqueous solution, alcohol solution with carbon chain ≤ 4, acetonitrile aqueous solution and methanol; in one embodiment of the present invention, it is methanol aqueous solution.

[0040] Preferably, the mobile phase used in the HPLC preparative separation is one or more of an acetone aqueous solution, an alcohol solution with a carbon chain ≤ 4, and an acetonitrile aqueous solution; in one embodiment of the present invention, it is an acetonitrile aqueous solution.

[0041] As a preferred embodiment of the present invention, the medium used in the gel column chromatography can be Sephadex LH-20, the mobile phase used can be methanol, and the liquid eluted with the mobile phase is named component L1-L 10 In the above liquid fermentation preparation, the preparation method includes separating sporechelin AE and sporechelin A1 from the component L3.

[0042] In the above-mentioned liquid fermentation preparation of spore chelating agent AE and spore chelating agent A1, the separation of spore chelating agent AE and spore chelating agent A1 from the component L3 includes subjecting the component L3 to reverse silica gel column chromatography, gradient eluting with 10%-100% methanol aqueous solution, and naming the liquid eluted with the mobile phase as L 3-1 -L 3-11 , component L 3-6 Sporochelin C and component L were also obtained by HPLC preparation and separation. 3-7 Sporochelin BD and component L were also obtained by HPLC preparation and separation. 3-8 Sporochelin BE and A1 were also obtained by HPLC preparation and separation, and component L 3-9 Sporochelin A is obtained by HPLC preparative separation. The mobile phase used in the HPLC preparative separation can be an acetonitrile and water system.

[0043] The present invention provides a catechol compound and a preparation method and application thereof. The catechol compound is a natural product obtained by microbial fermentation, has good anti-fibrosis and anti-inflammatory activities, and has good clinical application prospects for anti-fibrosis and anti-inflammatory treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] In order to more clearly illustrate the technical solutions in the present invention or the prior art, a brief introduction is given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0045] Figure 1 The compound represented by formula I-VI 1 H- 1 H COSY and HMBC related signals.

[0046] Figure 2 This is a high-resolution electrospray ionization mass spectrum of the compound represented by Formula I.

[0047] Figure 3 For the compound shown in formula I 1 H NMR spectrum.

[0048] Figure 4 For the compound shown in formula I 13 C NMR spectrum.

[0049] Figure 5 This is the high-resolution electrospray ionization mass spectrum of the compound represented by formula II.

[0050] Figure 6 The compound represented by formula II 1 H NMR spectrum.

[0051] Figure 7 The compound represented by formula II 13 C NMR spectrum.

[0052] Figure 8 This is the high-resolution electrospray ionization mass spectrum of the compound represented by formula III.

[0053] Figure 9 The compound represented by formula III 1 H NMR spectrum.

[0054] Figure 10 The compound represented by formula III 13 C NMR spectrum.

[0055] Figure 11 This is the high-resolution electrospray mass spectrum of the compound represented by formula IV.

[0056] Figure 12 The compound represented by formula IV 1 H NMR spectrum.

[0057] Figure 13 The compound represented by formula IV 13 C NMR spectrum.

[0058] Figure 14 This is a high-resolution electrospray ionization mass spectrum of the compound represented by formula V.

[0059] Figure 15 The compound represented by formula V 1 H NMR spectrum.

[0060] Figure 16 The compound represented by formula V 13 C NMR spectrum.

[0061] Figure 17 This is a high-resolution electrospray ionization mass spectrum of the compound represented by formula VI.

[0062] Figure 18 The compound represented by formula VI 1 H NMR spectrum.

[0063] Figure 19 The compound represented by formula VI 13 C NMR spectrum.

[0064] Figure 20 The electronic circular dichroism spectra of the compound represented by formula I and 2S / 2R-sporocytin A calculated by quantum chemistry are shown.

[0065] Figure 21 The electronic circular dichroism spectra of the compounds represented by formula I-VI are shown in FIG.

[0066] Figure 22 The cytotoxicity of sporechelin AE and sporechelin A1 in LX-2 cells.

[0067] Figure 23 The cytotoxicity of sporechelin AE and sporechelin A1 in NIH3T3 cells stimulated by TGF-β1.

[0068] Figure 24 Toxicity evaluation of sporechelin AE and sporechelin A1 in LPS-induced Raw264.7 cells (1 μM). DETAILED DESCRIPTION

[0069] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0070] Example 1

[0071] This embodiment provides a catechol compound and a preparation method thereof, comprising:

[0072] 1. Fermentation of Micromonospora sp.TMD166

[0073] The spore solution was inoculated onto a No. 38 culture medium plate. The culture medium formula was as follows: glucose 4.0g, yeast extract 4.0g, malt extract 5.0g, ZnSO4.7H2O 0.001g, MnCl2.4H2O0.001g, FeSO4.7H2O 0.002g, phenylalanine 0.001g, alanine 0.0003g, vitamin B1 0.001g, vitamin B2 0.001g, vitamin B6 0.001g, niacin 0.001g, biotin 0.001g, agar 15g, sterile distilled water 1.0L, pH 7.2. The plate was incubated at 28℃ for 7-10 days. A sterile bamboo stick was used to remove a 1cm 2 The agar block containing bacteria was inoculated into 100 mL of liquid No. 38 culture medium in a 500 mL Erlenmeyer flask and cultured at 28°C, 180 rpm, and shaken for 3-4 days to prepare the seed solution.

[0074] 100 mL of the above seed solution was transferred to a 5 L conical flask containing 1 L of No. 38 liquid culture medium, for a total of 30 L, and cultured in a shake flask at 28°C and 180 rpm for 6-7 days to obtain the fermentation product of Micromonospora sp. TMD166.

[0075] 2. Extraction of Micromonospora sp.TMD166 fermentation products and preparation of crude extracts

[0076] The liquid fermentation product (30 L) obtained above was centrifuged to obtain a supernatant, which was extracted three times with an equal volume of ethyl acetate (30 L×3). After concentration at low temperature and reduced pressure to remove the organic solvent, a crude ethyl acetate extract (3.2 g) was obtained.

[0077] 3. Isolation, Preparation and Structural Identification of Compounds Represented by Formulas I-VI

[0078] The crude ethyl acetate extract obtained above was dissolved in methanol and separated by gel column chromatography, with pure methanol solution as the mobile phase. The separation medium used in the gel column chromatography was Sephadex LH-20, the column size was 25×2400 mm (diameter×length), and the column volume was 1100 mL. From the beginning of the elution process, the eluted liquid was continuously collected, 15 mL was collected in each tube, and a total of 99 tubes were collected. The collected effluent was detected by TLC and combined to obtain 10 components (L1-L 10 ). Fraction L3 (732 mg, i.e., the collected solution from tubes 17 to 26) was purified by C18 Reversed phase flash column chromatography, C 18 The medium used in the reversed-phase silica gel column was column chromatography reversed-phase silica gel (50 g). The specifications of the reversed-phase silica gel column used were 45 × 90 mm (diameter × length). The elution program was 0-100 min, 10%-100% methanol, 100-120 min, 100% methanol, 15 mL / min. The eluted liquid was continuously collected from the beginning of the elution program, 20 mL was collected in each tube, and a total of 90 tubes were collected. They were combined according to the UV absorption graph to obtain 11 components (L 3-1 -L 3-11 ); component L 3-6 (32 mg, collected from tubes 33 to 38) was separated and purified by HPLC (chromatographic column parameters: Capcell-Pak-C8 DD, 5 μm, 10 mm×250 mm), and eluted with acetonitrile-water solution (acetonitrile and water volume ratio of 38:62) (flow rate of 2 mL / min) to obtain the compound of formula IV (4.6 mg, t R =25.7min); component L 3-7 (39 mg, collected from tubes 39 to 42) was separated and purified by HPLC (chromatographic column parameters: Capcell-Pak-C 18 MGⅡ, 5μm, 10mm×250mm), eluted with acetonitrile-water solution (acetonitrile and water volume ratio of 43:57) (flow rate of 2mL / min) to obtain the compound of formula III (9mg, t R =23.0min), the compound of formula Ⅳ (4 mg, t R =18.2min), the compound of formula V (6.3 mg, t R =25.7min); component L 3-8 (22 mg, collected from tubes 43 to 46) were separated and purified by HPLC (chromatographic column parameters: Capcell-Pak-C 18 MGⅡ, 5μm, 10mm×250mm), eluted with acetonitrile-water solution (acetonitrile and water volume ratio of 43:57) (flow rate of 2mL / min) to obtain the compound of formula Ⅱ (0.9mg, t R =22.1min), the compound of formula III (1.3 mg, t R =22.6min), the compound of formula Ⅳ (1.6mg, t R =18.3min), the compound of formula V (0.8mg, t R =25.4min), the compound of formula VI (5.6 mg, t R =36.0min), component L 3-9(62 mg, collected from tubes 47 to 57) were separated and purified by HPLC (chromatographic column parameters: Capcell-Pak-C 18 MGⅡ, 5μm, 10mm×250mm), eluted with acetonitrile-water solution (acetonitrile and water volume ratio of 45:55) (flow rate of 2mL / min) to obtain the compound of formula I (42mg, t R =44.3min).

[0079] Structural identification of the compound spore chelating agent A represented by formula I: High resolution electron spray mass spectrometry (HRESIMS) showed a quasi-molecular ion peak m / z 523.2081 [M+H] + , combined with nuclear magnetic resonance spectroscopy (NMR) data, its molecular formula was determined to be C 28 H 31 N2O8 (theoretical value is 523.2080, Figure 2 ), the degree of unsaturation is 15. The compound represented by formula I 1 H NMR spectra (CD3OD, Figure 3 , Table 1) shows 11 aromatic hydrogen proton signals in the low field region of 6.68-7.19 ppm, 5 methylene signals in the high field region of 1.46-3.61 ppm, and one oxymethylene signal δ H 4.21 (dd, H-1a), 4.15 (dd, H-1b), a methine signal δ H 4.38 (m, H-2). 13 C NMR spectra (CD3OD, Figure 4 , Table 2) showed 28 carbon signals, and DEPT and HSQC spectra showed that these carbons were 10 quaternary carbons (3 carbonyl carbons, 7 sp 2 quaternary carbon), 12 methine carbons [11 of which are sp 2 Hybridized carbon, one is nitrogen-bonded carbon δ C 49.6 (C-5')], 6 methylene carbons [one of which is an oxygen-linked carbon δ C 67.2 (C-1)]. The above NMR data are similar to those of Myxochelin A isolated from Myxobacteria (see Kunze B, Bedorf N, Kohl W, G, Reichenbach H. Myxochelin A, a new iron-chelating compound from Angiococcus disciformis (Myxobacterales). Production, isolation, physico-chemical and biological properties. J Antibiot[J]. 1989, 42(1): 14-17.), suggesting that the compound is a myxochelin derivative. The COSY spectrum of the compound of formula I shows that the structure contains the following peaks: Figure 1 The four structural fragments shown include a 2,6-diaminohexanol, a monosubstituted benzene ring, and two trisubstituted o-diphenol benzene rings. In the HMBC spectrum, the correlations of H-6" with C-7" and H-6' with C-7' indicate the presence of two o-diphenol benzoyl fragments in the structure. The correlations of H-6 with C-7" and H-2 with C-7' indicate that the two o-diphenol benzoyl fragments are linked to the 2,6-diaminohexanol via amide bonds, confirming the compound's mucochelin skeleton. Furthermore, the correlations of H-2"' with C-1", C-3", C-4", and C-8", combined with the chemical shifts of these carbons, indicate the presence of a phenylacetyl unit in the structure. The correlations of C-1 and C-1"' indicate that the phenylacetyl group is attached to the C-1 position via an ester bond, confirming the planar structure of the compound of Formula I.

[0080] Structural identification of the compound spore chelating agent A1 represented by formula II: HRESIMS showed a quasi-molecular ion peak m / z 492.2147 [M+H] + (C 27 H 30 The theoretical value of N3O6 is 492.2135, Figure 5 ), combined with NMR data, its molecular formula was determined to be C 27 H 29 N3O6, which is 31Da less than the molecular weight of the compound of formula I. The NMR data of the compound of formula II are very similar to those of the compound of formula I ( Figure 6 , Figure 7 ), the main difference is that the compound of formula II 1 A group of δ appears in H NMR H The hydrogen signals at 8.08, 8.65 and 8.84 ( Figure 6 ), indicating the presence of an aromatic heterocycle in the structure. HMBC and COSY analysis showed that the heterocycle was a nicotinyl group. The correlation between H-2 and C-7' showed that the nicotinyl group was attached to the C-2 position. Therefore, the structure of the compound was determined to be as shown in Formula II.

[0081] Table 1 1H NMR data of compounds represented by formula I-VI (CD3OD, 600 MHz)

[0082]

[0083] Table 2 13C NMR data of compounds represented by formula I-VI (CD3OD, 150 MHz)

[0084] serial number Sporochelin A Sporochelin A1 Sporochelin B Sporochelin C Sporochelin D Sporochelin E 1 <![CDATA[67.2,CH2]]> <![CDATA[67.2,CH2]]> <![CDATA[67.1,CH2]]> <![CDATA[67.2,CH2]]> <![CDATA[66.9,CH2]]> <![CDATA[66.8,CH2]]> 2 49.6,CH 50.3,CH 49.6,CH 49.7,CH 49.7,CH 49.7,CH 3 <![CDATA[31.5,CH2]]> <![CDATA[31.5,CH2]]> <![CDATA[31.5,CH2]]> <![CDATA[31.6,CH2]]> <![CDATA[31.6,CH2]]> <![CDATA[31.6,CH2]]> 4 <![CDATA[24.3,CH2]]> <![CDATA[24.3,CH2]]> <![CDATA[24.3,CH2]]> <![CDATA[24.4,CH2]]> <![CDATA[24.4,CH2]]> <![CDATA[24.4,CH2]]> 5 <![CDATA[30.1,CH2]]> <![CDATA[30.1,CH2]]> <![CDATA[30.1,CH2]]> <![CDATA[30.1,CH2]]> <![CDATA[30.1,CH2]]> <![CDATA[30.1,CH2]]> 6 <![CDATA[40.2,CH2]]> <![CDATA[40.1,CH2]]> <![CDATA[40.2,CH2]]> <![CDATA[40.2,CH2]]> <![CDATA[40.2,CH2]]> <![CDATA[40.2,CH2]]> 1′ 116.8，C 116.8，C 116.9，C 116.8，C 116.8，C 2′ 150.2，C 149.1, CH 150.2，C 150.1，C 150.3，C 150.1，C 3′ 147.3，C 132.0,C 147.3，C 147.3，C 147.3，C 147.3，C 4′ 119.6,CH 137.0, CH 119.6, CH 119.7, CH 119.6,CH 119.7, CH 5′ 119.5, CH 125.1, CH 119.5, CH 119.5, CH 119.5, CH 119.5, CH 6′ 118.6, CH 152.5, CH 118.6, CH 118.9, CH 118.6, CH 118.8, CH 7′ 171.5，C 168.0，C 171.5，C 171.5,C 171.5，C 171.5，C 1″ 116.8，C 116.7，C 116.8，C 116.8，C 116.9，C 116.7，C 2″ 150.2，C 150.3,C 150.1，C 150.3，C 150.1，C 150.2，C 3″ 147.3，C 147.3，C 147.3，C 147.3，C 147.3，C 147.3，C 4″ 119.7, CH 119.5,CH 119.7, CH 119.6,CH 119.6, CH 119.6,CH 5″ 119.5,CH 119.5, CH 119.5,CH 119.5,CH 119.5,CH 119.5,CH 6" 118.9,CH 118.5,CH 118.9, CH 118.6,CH 118.9,CH 118.6, CH 7″ 171.5，C 171.5,C 171.5,C 171.5,C 171.5,C 171.5,C 1″′ 173.4,C 173.3,C 173.9,C 172.8，C 176.1,C 178.6,C 2″′ <![CDATA[42.0,CH2]]> <![CDATA[42.0,CH2]]> <![CDATA[41.1,CH2]]> <![CDATA[20.7,CH3]]> <![CDATA[28.2,CH2]]> 35.2,CH 3″′ 135.4,C 135.5,C 126.2，C <![CDATA[9.4,CH3]]> <![CDATA[19.2,CH3]]> 4″′ 130.3,CH 130.3,CH 131.3,CH <![CDATA[19.3,CH3]]> 5″′,7″′ 129.5,CH 129.5, CH 116.3, CH 6″′ 128.0,CH 128.0, CH 157.5，C 8″′ 130.3,CH 130.3,CH 131.3,CH

[0085] Structural identification of the compound represented by formula III: HRESIMS showed a quasi-molecular ion peak m / z 539.2021 [M+H] + (C 28 H 31 The theoretical value of N2O9 is 539.2030, Figure 8 ), combined with NMR data, its molecular formula was determined to be C 28 H 30 N2O9 has one more oxygen atom than the compound represented by formula I. The NMR data of the compound represented by formula III are very similar to those of the compound represented by formula I ( Figure 9 , Figure 10 ), the main difference is in the phenylacetyl structural fragment. 13 C NMR has an additional oxygen-linked aromatic carbon δ C 157.5, in addition, C-3″′, C-5″′, and C-7″′ shifted to -9.2, -13.2, and -13.2 ppm upfield, respectively ( Figure 10 ), suggesting that H-6"' of the compound of formula I is substituted by OH. 2D NMR data analysis further confirmed that the compound of formula III is the C-6"'-hydroxylation product of the compound of formula I.

[0086] Structural identification of the compound represented by formula IV: HRESIMS showed a quasi-molecular ion peak m / z 447.1786 [M+H] + (C 22 H 27 The theoretical value of N2O8 is 447.1767, Figure 11 ), combined with NMR data, its molecular formula was determined to be C 22 H 26 N2O8, compared with the NMR data of the compound of formula I, the compound of formula IV lacks a group of phenethyl signals ( Figure 12 , Figure 13 ), 1 H NMR has an additional singlet methyl signal δ H2.02, and the methyl group and C-1"' have related signals in the HMBC spectrum. This suggests that the phenylacetyl group in the structure of formula I is replaced by an acetyl group. The above speculation is confirmed by 2D NMR data analysis, and the structure of the compound of formula IV is determined.

[0087] The compound represented by Formula V, spore chelating agent D, is a white powder that is easily soluble in methanol, chloroform, and DMSO. High-resolution electron spray mass spectrometry (HRESIMS) shows a quasi-molecular ion peak of m / z 461.1914 [M+H] + (C 23 H 29 The theoretical value of N2O8 is 461.1924, Figure 14 ), combined with NMR data, its molecular formula was determined to be C 23 H 28 N2O8. The NMR data of the compound of formula V is very similar to that of the compound of formula IV, except that 1 H and 13 C NMR has an additional methylene signal (δ H 2.32, δ C 28.2)( Figure 15 , Figure 16 ), and the methyl carbon signal in the compound of formula IV shifts 11.3 ppm to the upfield, and C-1"' shifts 3.3 ppm to the downfield, and 1 H- 1 H-2"'(q,δ H 2.32) and H-3"'(t,δ H 1.07) and the correlation between H-2"' and C-1"' (δ C The correlation between the results of the above analysis indicated that the acetyl group in the structure of formula IV was replaced by a propionyl group, and the structure of the compound shown in formula V was determined and named as sporophore D.

[0088] Structural identification of the compound spore chelating agent E represented by formula VI: High-resolution electron spray mass spectrometry (HR-ESI-MS) showed a quasi-molecular ion peak of m / z 475.2063 [M+H] + (C 24 H 31 The theoretical value of N2O8 is 475.2080, Figure 17 ), combined with NMR data, its molecular formula was determined to be C 24 H 30 N2O8, has one more methylene group than the compound of formula V. Compared with the NMR data of the compound of formula V, the compound of formula VI 1 H and 13 C NMR has one less ethyl signal and one more methine signal (δ H 2.53, δ C35.2) and two doublets of methyl signals (δ H 1.09,1.10δ C 19.2,19.3), and C-1"' shifted 2.5ppm downfield ( Figure 18 , Figure 19 ). Through 2D NMR data analysis, it was found that the two newly appeared methyl groups were respectively H 2.53, δ C 35.2) in COSY spectrum and are correlated with C-1"' in HMBC spectrum, indicating that the propionyl group in the compound of formula V is replaced by isobutyryl to obtain the compound of formula VI.

[0089] The absolute configuration of the compound of formula I was determined by electron circular dichroism (ECD) calculation and literature comparison. The measured circular dichroism spectrum of the compound of formula I was fitted with the electron circular dichroism spectra of the quantum calculated S-1a and R-1b configuration compounds. The compound of formula I showed a negative cotton effect at 220nm and 260nm, and a positive cotton effect at 240nm and 310nm, which was the same as the electron circular dichroism spectrum of the S-configuration compound (1a) calculated by quantum chemistry ( Figure 20 ), the absolute configuration of the compound of formula I is determined to be 2S. In addition, the specific rotation value of the compound of formula I is [α] 20 D -8.8 (c 0.5, MeOH). The literature reports that the specific rotation value of the synthesized S-configuration myxochelin A is also negative (see Furumai T, Igarashi Y. Absolute configuration and antitumor activity of myxochelin A produced by Nonomuraeapusilla TP-A0861. J. Antibiot. 2006, 59, 698-703.), further confirming that the chiral center C-2 of the compound of formula I is S-configuration. The specific rotation values ​​of the compounds represented by formula II-VI are all negative, and the ECD spectra are similar to those of the compound of formula I ( Figure 21 ), therefore, the absolute configuration of the compound represented by formula II-VI is also 2S.

[0090] The physicochemical properties of the compounds shown above:

[0091] Formula I compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -8.8(c0.5,MeOH);UV(MeOH)λ max(logε)207(4.97),249(4.36),312(3.91)nm; ECD(c 2.39×10 -4 M)λ max (Δε)203(0.43),218(-3.20),240(+1.46),261(-1.10),314(+0.42)nm; 1 H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 523.2081 [M+H] + (C 28 H 31 The theoretical value of N2O8 is 523.2080).

[0092] Formula II compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -4.8(c0.25,MeOH);UV(MeOH)λ max (logε)207(4.27),248(3.69),313(3.15)nm; ECD(c 1.22×10 -4 M)λ max (Δε)207(-2.05),216(-3.55),240(-1.45),317(+0.32); 1 H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 492.2147 [M+H] + (C 27 H 30 The theoretical value of N3O6 is 492.2135).

[0093] Formula III compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -8.0(c0.5,MeOH);UV(MeOH)λ max (logε)206(4.96),249(4.39),314(3.94)nm; ECD(c 2.32×10 -4 M)λ max (Δε)204(+1.52),220(-2.10),240(+1.57),260(-1.18),310(+0.31)nm; 1H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 539.2021 [M+H] + (C 28 H 31 The theoretical value of N2O9 is 539.2030).

[0094] Formula IV compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -8.8(c1.0,MeOH);UV(MeOH)λ max (logε)207(4.91),250(4.37),314(3.93)nm; ECD(c 2.80×10 -4 M)λ max (Δε)207(0.09),218(-0.87),239(+1.26),260(-1.37),314(+0.53); 1 H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 447.1786 [M+H] + (C 22 H 27 The theoretical value of N2O8 is 447.1767).

[0095] Formula V compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -4.0(c0.1,MeOH);UV(MeOH)λ max (logε)207(4.82),249(4.28),314(3.84)nm; ECD(c 2.72×10 -4 M)λ max (Δε)207(+0.02),215(-0.71),240(+0.81),263(-0.76),315(+0.41); 1 H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 461.1914 [M+H]+ (C 23 H 29 The theoretical value of N2O8 is 461.1924).

[0096] Formula VI compound: white powder, easily soluble in methanol, chloroform, and DMSO. [α] 20 D -6.4(c0.5,MeOH);UV(MeOH)λ max (logε)207(4.91),249(4.37),314(3.92)nm; ECD(c 2.64×10 -4 M)λ max (Δε)207(-0.96),214(-1.48),240(+0.90),264(-1.01),314(+0.38)nm; 1 H-NMR (CD3OD, 600MHz) data are shown in Table 1. 13 C NMR (CD3OD, 150 MHz) data are shown in Table 2; HRESMS: m / z 475.2063 [M+H] + (C 24 H 31 The theoretical value of N2O8 is 475.2080).

[0097] 4. In vitro anti-fibrotic activity and cytotoxicity evaluation test of sporechelin AE and sporechelin A1: This part uses human hepatic stellate cells LX-2 and mouse embryonic fibroblasts NIH3T3 to evaluate the anti-fibrotic effect of the compounds. The former is a commonly used model for screening anti-fibrotic drugs, and the latter is widely used in the study of cell models of organ fibrosis induced by tissue hypoxia such as pulmonary fibrosis, cardiovascular fibrosis, liver fibrosis, renal fibrosis, skin fibrosis and clinical interventional embolization.

[0098] Cell culture and compound treatment: Human hepatic stellate cells LX-2 were cultured in DMEM medium (Gibco) containing 10% fetal bovine serum and 1% penicillin-streptomycin (Gibco) at a concentration of 3 × 10 4 / cm 2 Cells were plated in 96-well plates at a density of 20 μg / mL and incubated in a 37°C, 5% CO2 incubator until 60-80% confluency was achieved. After that, serum-free DMEM medium was replaced with starvation medium for 24 hours. Subsequently, the original medium was discarded and treated for 24 hours with TGF-β1 at a final concentration of 2 ng / mL, 5 μM and 1 μM sporechelin AE and sporechelin A1, or 20 μM of the positive agent EGCG (epigallocatechin). A control group (no TGF-β1 or compound) and a TGF-β1-induced group (only TGF-β1) were also established. Mouse embryonic fibroblasts (NIH3T3) were plated and cultured similarly to LX-2 cells. After culturing to 60-80% confluency, 5 ng / mL TGF-β1 and the aforementioned compounds were added and treated for 24 hours.

[0099] Total RNA extraction and detection of fibrosis marker gene expression: The supernatant of the cell culture medium in the 96-well plate was discarded, and total cell RNA was extracted using a total RNA extraction kit (Magen) according to the operating instructions. A 10 μL reaction system was prepared according to the instructions of the HiScript II One Step qRT-PCR SYBR Green Kit (Vazyme), and an ABI 7500Fast instrument was used to perform a one-step qRT-PCR method to quantitatively detect the mRNA expression of human fibrosis marker genes COL1A1, ACTA2, TGFB1 and mouse fibrosis marker genes Col1a1, Acta2. The internal reference gene was GAPDH (Gapdh). The primer sequences used are shown in Table 3. Table 3 qRT-PCR primer sequences

[0100]

[0101] Compared with the model group, the results of the inhibition rates of sporechelin AE and sporechelin A1 on the mRNA expression of fibrosis marker genes are shown in Tables 4 and 5:

[0102] Table 4 Inhibition rate of fibrosis gene mRNA expression by sporechelin AE and sporechelin A1 in LX-2 cells (%)

[0103]

[0104] Table 5 Inhibition rate of fibrosis gene mRNA expression by sporechelin AE and sporechelin A1 in NIH3T3 cells (%)

[0105]

[0106] Cytotoxicity evaluation of spore sequestrant AE and spore sequestrant A1: Human hepatic stellate cells LX-2 were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at a rate of 3×10 4 / cm 2 Cells were plated in a 96-well plate at a density of 50-80% at 37°C in a 5% CO2 cell culture incubator. Then, different concentrations of spore chelating agent AE and spore chelating agent A1 were added for 24 hours. Experimental groups (compounds added to cells), control groups (DMSO solvent added to cells) and blank groups (culture medium only) were set up. Subsequently, the culture medium was discarded, and a mixture of 90 μL of fresh culture medium and 10 μL of CCK-8 was added to each well. The cells were cultured in a 37°C 5% CO2 cell culture incubator for 1-3 hours. The absorbance at 450 nm (OD value) was read using a microplate reader. Cell survival rate = [(experimental group - blank group) / (control group - blank group)] × 100%. Cell survival rate was calculated as follows: Figure 22Similarly, after NIH3T3 cells were stimulated with 5 ng / mL TGF-β1 and treated with 5 μM compound for 24 hours, their cytotoxicity was evaluated. The results are shown in Figure 23 shown.

[0107] 5. Evaluation of the Inhibition of 5-Lipoxygenase Activity by Sporochelin AE and Sporochelin A1 in a Cell-Free System. This assay used 5-Lipoxygenase active protein in vitro under cell-free conditions to assess the inhibitory effects of compounds on its activity. The 5-Lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), was used as a positive agent. The assay was performed as follows: 1 μL of compound, DMSO solvent, or NDGA positive agent was pre-added to a 96-well plate and mixed with 50 μL of enzyme assay solution (containing purified 5-LOX active protein and assay solution). The plate was then incubated at 37°C for 15 minutes. Subsequently, 50 μL of reaction buffer (containing 6 μM arachidonic acid, 20 μM ATP, and 10 μM H2DCFDA) was added. After 30 minutes, fluorescence intensity was measured at 485 / 535 nm (excitation / emission wavelengths). The inhibitory activity of the compound against 5-Lipoxygenase was calculated based on the fluorescence intensity of each group. The results are shown in Table 6. The inhibition rates of the above compounds on 5-lipoxygenase activity at a concentration of 2 μM were all >50%:

[0108] Table 6 Inhibition rate of 5-lipoxygenase activity by sporechelin AE and sporechelin A1 in the cell-free system (%)

[0109] Compound (2 μM) 5-lipoxygenase activity inhibition rate (%) Sporochelin A 72.45±13.30 Sporochelin A1 66.00±17.51 Sporochelin B 76.85±13.79 Sporochelin C 78.31±18.61 Sporochelin D 74.77±12.38 Sporochelin E 79.63±23.41 NDGA 69.68±8.09

[0110] 6. In vitro anti-inflammatory activity evaluation test of spore sequestrant AE and spore sequestrant A1: This part uses bacterial lipopolysaccharide (LPS) to induce mouse peritoneal macrophage Raw264.7 inflammatory response model to evaluate the anti-inflammatory effect of the compounds. This model is a cell model widely used in various inflammatory injury diseases, and is also a commonly used model for anti-inflammatory drug screening and efficacy evaluation.

[0111] Cell culture and compound treatment: Raw264.7 cells were cultured in DMEM medium (Gibco) containing 10% fetal bovine serum and 1% penicillin-streptomycin (Gibco) at a rate of 3 × 10 4 / cm 2The cells were plated in a 96-well plate at a density of 50 μL and cultured in a 37°C 5% CO2 cell culture incubator until 60-80% was reached. 50 μL of spore-chelating agent AE, spore-chelating agent A1 and positive drug dexamethasone (Dex) were added for pretreatment for 2 hours. Subsequently, 50 μL of 1 μg / mL LPS was added to make the final concentrations of spore-chelating agent AE and spore-chelating agent A1 1 μM and the final concentration of Dex 5 μM. Total RNA was extracted after 18 hours of induction (the method was the same as before). The mRNA expression of pro-inflammatory factors Tnfα and Cox2 genes was detected by qRT-PCR. At the same time, the same conditions were set to evaluate their cytotoxicity using the CCK-8 method. The primer sequences used are shown in Table 3. Compared with the model group, the inhibition rates of spore-chelating agent AE and spore-chelating agent A1 on the mRNA expression of inflammatory response marker genes and the cytotoxicity results on Raw264.7 under this condition are shown in Table 7 and Table 8. Figure 24 As shown:

[0112] Table 7 Inhibition rate of sporechelin AE and sporechelin A1 on mRNA expression of inflammatory response marker genes (%)

[0113]

[0114] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A catechol compound, characterized in that Having any of the following structural formulas: Wherein, R1 represents methyl, ethyl, isopropyl, benzyl or p-hydroxybenzyl; R2 represents benzyl.

2. Use of the catechol compound according to claim 1 in the preparation of anti-fibrosis drugs.

3. Use of the catechol compound according to claim 1 in the preparation of anti-inflammatory drugs.

4. An anti-fibrosis drug, characterized in that: Contains the catechol compound according to claim 1 or a pharmaceutically acceptable salt thereof.

5. An anti-inflammatory drug, characterized in that Contains the catechol compound according to claim 1 or a pharmaceutically acceptable salt thereof.

6. The method for preparing catechol compounds according to claim 1, characterized in that: The method comprises S1 and S2, wherein S1: fermenting and culturing Micromonospora to obtain fermentation liquid; and S2: extracting and separating the catechol compound from the fermentation liquid; the Micromonospora is Micromonospora sp.TMD166.

7. The method for preparing catechol compounds according to claim 6, characterized in that: The S2 includes S21 to S22, S21: centrifuging the fermentation broth to obtain a supernatant and mycelium, taking the supernatant for extraction treatment to obtain a supernatant extract; S22: preliminarily separating the supernatant extract through gel column chromatography, then separating it through reverse-phase silica gel column chromatography, performing gradient elution with a solvent, and finally using HPLC to prepare and separate to obtain catechol compounds.

8. The method for preparing catechol compounds according to claim 7, characterized in that: During the extraction treatment, the extraction solvent is one or more of chloroform, dichloromethane, ethyl acetate and acetone.

9. The method for preparing catechol compounds according to claim 7, characterized in that: The medium used in the gel column chromatography separation is Sephadex LH-20 or Tosoh Toyopearl HW-40; and / or, the medium used in the reversed-phase silica gel column is C18 reversed-phase silica gel; and / or, the chromatographic column used in the HPLC preparative separation is a C18 chromatographic column, a C8 chromatographic column, a C3 chromatographic column or a phenyl bonded column.

10. The method for preparing catechol compounds according to claim 9, characterized in that: The mobile phase of the gel column chromatography is one or more of chloroform, dichloromethane, ethyl acetate, alcohols with a carbon chain of ≤4, acetone and butanone; or a mixed solution of one or more of the above solvents and water.

11. The method for preparing catechol compounds according to claim 9, characterized in that: The solution used for the reversed-phase silica gel column chromatography separation is one or more of an acetone aqueous solution, an alcohol solution with a carbon chain of ≤4, and an acetonitrile aqueous solution.

12. The method for preparing catechol compounds according to claim 9, characterized in that: The mobile phase used in the HPLC preparative separation is one or more of an acetone aqueous solution, an alcohol solution with a carbon chain of ≤4, and an acetonitrile aqueous solution.

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