A halenaquinol derivative and its application in anti-marine biofouling

Abstract

The present application relates to the technical field of marine antifouling, in particular to a bruceanol derivative and its application in resisting marine biofouling, using a directional multi-target ligand strategy, one of the main compounds in the widely sourced plant brucea javanica, bruceanol, is structurally modified, and strong marine source antifouling active groups (halogen, cyano, benzene ring, heterocycle, etc.) are hybridized on the bruceanol mother nucleus molecule, a series of new hybrid marine source antifouling active group bruceanol derivatives are successfully synthesized. These compounds show good anti-barnacle attachment activity in barnacle anti-attachment experiments, and have low toxicity to target organisms. The method enriches the types of candidate compounds of antifouling agents, provides a new way for developing new and efficient antifouling coatings, and has good application prospect and industrial value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of marine antifouling, and particularly relates to a jacobiin derivative and application thereof in resisting marine biofouling. BACKGROUND

[0002] Marine biofouling refers to the attachment and growth of marine organisms such as barnacles, bryozoa and sponges on man-made structures such as ships and offshore platforms, and causes physical and chemical damage to these structures. Since the mid-20th century, the use of metal ions and biocides to make antifouling coatings to reduce or eliminate fouling organisms on the surface of offshore facilities is the most economical and feasible method. However, these practices pose a great threat to the environment. The release of harmful chemicals by toxic coatings can also harm the normal growth and development of non-target organisms, and accumulate biological toxins through the food chain, ultimately endangering human health. Therefore, there is an urgent need to find new types of antifouling agents that are efficient, low-toxic and environmentally friendly to replace traditional antifouling agents. Natural products not only can effectively prevent and control fouling organisms such as barnacles, but also are friendly and safe to the environment, which is a potential natural molecule library for exploring new antifouling agents, and an important material basis for subsequent independent innovation and development of new antifouling coatings.

[0003] Natural products from marine sources have long played an important role in antifouling research, thanks to the ability of a wide variety of marine organisms to synthesize a series of natural compounds with unique structures and significant activity. Many marine natural products show excellent activity in preventing the attachment of fouling organisms such as barnacles. However, land-derived natural products also have unique advantages, such as wide sources, easy access, easy artificial cultivation, high yield and low production cost. A variety of land-derived natural products also show excellent antifouling activity, such as amaroid, capsaicin, cypermethrin, juvenile hormone, terpenes, cyclic peptides, alkaloids, flavonoids, tannins and other phenolic compounds, coumarin, polyketides and dihydro stilbenes, providing an ideal choice for the sustainable development of antifouling agents. Therefore, further use of the directed multi-target ligand strategy on the abundant land-derived antifouling active core molecules to hybridize the strong antifouling active groups (halogen, cyano, benzene ring, heterocycle, etc.) from marine sources to construct a variety of synergistic derivatives will open up a new window for the exploration of new natural antifouling agents, and its application prospect is immeasurable. SUMMARY

[0004] In view of the above, it is necessary to provide a land-derived jacobiin hybrid derivative and its application in resisting marine biofouling, which enriches the types of candidate compounds for antifouling agents and provides a new way for the development of new and efficient antifouling coatings, and has good application prospect and industrial value.

[0005] To achieve the above object, the technical scheme adopted by the present application is:

[0006] A hybrid ocean source strong fouling active group quassine chaulmoogryl alcohol derivative or its salt, the specific structure is shown in general formula (I):

[0007]

[0008] In which R is the marine source of anti-fouling active group including halogenated alkyl, halogenated benzene ring, heterocyclic ring and the like.

[0009] The above-mentioned quassine chaulmoogryl alcohol derivative or its pharmaceutically acceptable salt is used for marine anti-fouling.

[0010] The present application provides a kind of new type of antifouling agent for preventing and treating marine biofouling using the chaulmoogryl alcohol derivative.

[0011] The present application provides a kind of preparation method of chaulmoogryl alcohol derivative, the method comprises the following steps:

[0012] (1) chaulmoogryl alcohol is reacted with tert-butyl dimethyl silyl trifluoromethanesulfonate under the catalysis of imidazole to generate 3-OH silyl protection intermediate B2;

[0013] (2) intermediate B2 is hydrolyzed under the catalysis of NaOH, and 15 side chain occurs hydrolysis, and hydrolysis product B3 containing 15-OH is obtained;

[0014] (3) hydrolysis product B3 is reacted with acyl chloride reagent or acid reagent under the catalysis of triethylamine, and intermediate product is obtained, and then the desilicon reaction of TBAF is used to obtain the final target product.

[0015] In the present application, further, the reaction temperature in the step (1) is 5-25 DEG C, and the reaction time is 12h;The reaction temperature of the step (2) is 0 DEG C, and the reaction time is 8h;The reaction temperature of the step (3) is 5-25 DEG C, and the reaction time is 8h.

[0016] The reaction general formula of the above preparation method is as follows:

[0017]

[0018] Based on the above preparation method, compounds 1-15 can be prepared;

[0019] Specifically, the preparation method of the first type of chaulmoogryl alcohol derivative compound 1-7 is as follows:

[0020] A) The intermediate B2 is obtained by reacting the bruceanol (B1) with tert-butyldimethylsilyl trifluoromethanesulfonate (TBSOTf) under the catalysis of imidazole, and the purpose of this step is to protect the 3-OH which is more active; the reaction temperature of this step is 5-25°C, and the reaction time is 12h;

[0021] B) The hydrolysis product B3 containing 15-OH is obtained by hydrolyzing the side chain at position 15 of the intermediate B2 under the catalysis of NaOH; the reaction temperature of this step is 0°C, and the reaction time is 8h;

[0022] C) The intermediate product is obtained by reacting the hydrolysis product B3 with an acyl chloride reagent under the catalysis of triethylamine (Et3N), and then the final target product is obtained by desilylation of TBAF; the reaction temperature of this step is 5-25°C, and the reaction time is 8h; different target products are obtained by reacting B3 with different acyl chloride reagents, and are denoted as compounds 1-7.

[0023] The reaction equation corresponding to the above steps is as follows:

[0024]

[0025]

[0026] The present application provides the second type of bruceanol derivative compounds 8-11 and compounds 12-14, and the preparation method comprises the following steps:

[0027] a) First, activate different types of carboxylic acids by using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) to form an activated intermediate state, and the reaction temperature of this step is 5-25°C, and the reaction time is 1h; the activated intermediate state is used to attack the 15-OH of the hydrolysis product B3 under the catalysis of DMAP, and then the target product 8-11 is successfully obtained at position 15 by desilylation, and is denoted as compounds 8-11, and the reaction temperature of this step is 5-25°C, and the reaction time is 0.5h.

[0028] b) First, activate different types of carboxylic acids by using N,N'-diisopropylcarbodiimide (EDCI) and 1-hydroxybenzotriazole (HOBT) to form an activated intermediate state, and the reaction temperature of this step is 5-25°C, and the reaction time is 1h; the activated intermediate state is used to attack the 15-OH of the hydrolysis product B3 under the catalysis of DMAP, and then the target product 12-14 is successfully obtained at position 15 by desilylation, and is denoted as compounds 12-14, and the reaction temperature of this step is 5-25°C, and the reaction time is 0.5h.

[0029] The reaction equation corresponding to the above steps is as follows:

[0030]

[0031] The present invention provides a method for preparing a third class of brucein derivative compound 15, the method comprising the following steps:

[0032] First, 4-aminoindole was activated using N,N-carbonyldiimidazole (CDI) to form an activated intermediate under the catalysis of triethylamine. The activated intermediate was then used to attack the 15-OH group of the hydrolysis product B3. Under the catalysis of DMAP, a desilication reaction was then carried out to successfully obtain the target product 15 at the 15 position, which was recorded as compound 15.

[0033] The reaction equations corresponding to the above steps are as follows:

[0034]

[0035] The present invention has the following beneficial effects:

[0036] This invention proposes for the first time the application of brucein derivatives containing hybrid marine-derived strong active groups in anti-marine biofouling. Brucein is a characteristic natural quassinolide structure derived from the terrestrial plant Brucea javanica. In previous studies, the applicants found that it has significant anti-barnacle adhesion activity. Building on this discovery, this study further utilized a targeted multi-target ligand strategy to structurally modify brucein, incorporating strong marine-derived antifouling active groups into a rich variety of synergistic derivatives to enhance their antifouling properties. Based on existing research results, the present invention significantly improves the antifouling activity of bruceiol through structural optimization, verifies the great potential of natural products from terrestrial plants in the development of antifouling agents, provides technical support for the research and development of new environmentally friendly antifouling agents, and supplements and expands the antifouling technology system based on marine natural products. Specifically, the present invention uses bruceiol, an active ingredient in Brucea javanica, as a raw material to synthesize several bruceiol derivatives through an economical and easy method. The compounds have good antifouling activity. The method has a simple process, is suitable for large-scale production, has a reliable and stable source, and has great potential for promotion and application. This method can greatly expand its application prospects and provide guidance for the screening of antifouling compounds from terrestrial sources. Antifouling technology system based on natural products from marine sources.

Brief Description of the Drawings

[0037] Figure 1 The inhibitory effect of brusatol derivatives on the attachment of barnacle (Balanus Amphitrite) larvae.

[0038] SeaNine 211 was used as a positive control. 50 / EC 50 The value represents the toxicity level, LC 50 / EC50 A ratio greater than 15 indicates lower toxicity. p<0.05(*), p<0.01(**), p<0.001(***), and p>0.05(ns).

[0039] Figure 2 These are the results of a resuscitation experiment on the larvae of the barnacle (Balanus amphitrite). [Specific implementation method]

[0040] To make the above-mentioned objects, features, and advantages of the present invention more readily apparent, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings. The following description sets forth numerous specific details to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways than those described herein, and those skilled in the art may make similar modifications without departing from the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0041] Example 1: Preparation of Intermediate B2

[0042] Bruceol B1 (200 mg, 0.384 mmol) was dissolved in N,N-dimethylformamide (0.8 mL) solution, followed by the addition of tert-butyldimethylsilyl trifluoromethanesulfonate (2-5 equivalents) and imidazole (5-8 equivalents), and stirred at room temperature for 12 hours under nitrogen protection. Among them, the conditions of adding 4 equivalents of tert-butyldimethylsilyl trifluoromethanesulfonate and 8 equivalents of imidazole are the optimal reaction conditions. Subsequently, extraction was carried out with ethyl acetate and water, and a crude extract was obtained by recrystallization from ethanol and water. The introduction of the silane group greatly reduced the water solubility of the intermediate B2. Utilizing this change in solubility, this example used ethanol and water recrystallization for the first time to purify the intermediate B2. Compared with traditional column chromatography and liquid chromatography separation methods, the purification efficiency of ethanol-water recrystallization was significantly increased and the cost was significantly reduced. The precipitate was collected by filtration. The crude extracts were further purified by silica gel column chromatography using a mixture of ethyl acetate and petroleum ether (1:1) as eluent to obtain the target product, intermediate B2, as a white powdery solid in a yield of 87%.

[0043] The intermediate B2 obtained by column chromatography was analyzed using conventional nuclear magnetic resonance spectroscopy and mass spectrometry, and the analysis results are as follows:

[0044] 1H NMR (600 MHz, Chloroform-d) δ 6.25 (s, 1H, H-15), 4.78 (s, 1H, H-7), 4.72 (d, J = 7.9 Hz, 1H, H-20ax), 4.26 (d, J = 4.2 Hz, 1H, H-11), 4.19 (s, 1H, H-12), 3.79 (dd, J = 7.9 Hz, 1H, H-20eq), 3.78 (s, 3H, 17-OCH3), 3.13 (s, 1H, H-14), 2.93 (d, J = 12.9 Hz, 1H, H-5), 2.89 (d, J = 15.8 Hz, 1H, H-lax), 2.38 (dt, J = 14.7, 2.9 Hz, 1H, H-6ax), 2.33 (d, J = 15.8 Hz, 1H, H-leq), 2.18 (d, J = 1.3 Hz, 3H, 5'-CH3), 2.07 (br s, 1H, H-9), 1.92 (d, J = 1.4 Hz, 3H, 4'-CH3), 1.85 (d, J = 1.7 Hz, 3H, 18-CH3), 1.75 (ddd, J = 14.6, 13.0, 2.7 Hz, 1H, H-6eq), 1.38 (d, J = 1.1 Hz, 3H, 19-CH3), 0.94 (s, 9H, H-Me3CSi), 0.17 (s, 3H, H-Me2Si), 0.13 (s, 3H, H-Me2Si). 13 CNMR (151 MHz, Chloroform-d) δ 192.4 (C-2), 172.3 (C-21), 167.5 (C-16), 164.8 (C-l'), 161.3 (C-3'), 145.4 (C-3), 136.2 (C-4), 114.4 (C-2'), 82.9 (C-7), 81.7 (C-13), 76.1 (C-12), 74.4 (C-20), 71.3 (C-11), 66.1 (C-15), 53.4 (17-OCH3), 51.9 (C-14), 50.9 (C-l), 45.7 (C-8), 43.0 (C-5), 42.3 (C-9), 40.8 (C-10), 29.7 (C-6), 28.0 (C-4'), 26.4 (Me3CSi, 3C), 21.0 (C-5'), 19.2 (Me3CSi, C), 15.8 (19-CH3), 14.7 (18-OCH3), -3.4 (Me2Si), -3.6 (Me2Si), MS (ESI): m / z = 635.39 [M+H] + , 652.42 [M+NH4] + , 657.38 [M+Na] +Anal. calcd. for C 32 H 46 O 11 Si (634.28 g / mol).

[0045] Example 2: Preparation of hydrolysis product B3

[0046] Intermediate B2 (100 mg, 0.1577 mmol) was dissolved in methanol (0.5 mL) and hydrolyzed by adding a base at 0 °C in a water bath, where the base can be sodium methoxide, potassium methoxide, sodium hydroxide, lithium hydroxide, potassium tert-butoxide, potassium carbonate, and the reaction equivalent range is 0.5-2, and the optimal reaction condition is 1.5 equivalent of sodium hydroxide. The mixture was continuously stirred at this temperature for 8 hours, and the reaction was stopped when the TLC detection showed that the reaction was complete, and the remaining sodium hydroxide was neutralized to neutral pH with dilute hydrochloric acid. Subsequently, extraction was performed with ethyl acetate and water, dried over anhydrous sodium sulfate, and concentrated in vacuum. The crude product was purified by silica gel column chromatography using a mixture of ethyl acetate-petroleum ether (7:3) as the eluent, and finally hydrolysis product B3 was obtained as a white powdery solid, with a yield of 84%.

[0047] The hydrolysis product B3 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectroscopy and mass spectrometry methods, and the analysis results are as follows:

[0048] 1 H NMR (600 MHz, Chloroform-d) δ 5.28 (d, J = 14.7, 2.9 Hz 1H, H-15), 4.73 (s, 1H, H-7), 4.71 (d, J = 7.9 Hz, 1H, H-20ax), 4.24 (br s, 1H, H-11), 4.21 (s, 1H, H-12), 3.78 (d, J = 7.9 Hz, 1H, H-20eq), 3.78 (s, 3H, 17-OCH3), 3.03 (s, 1H, H-14), 2.86 (d, J = 12.9 Hz, 1H, H-5), 2.91 (d, J = 15.8 Hz, 1H, H-1ax), 2.38 (dt, J = 14.7, 2.9 Hz, 1H, H-6ax), 2.28 (d, J = 15.8 Hz, 1H, H-1eq), 2.02 (d, J = 4.61 Hz, 1H, H-9), 1.85 (d, J = 1.7 Hz, 3H, 18-CH3), 1.77 (ddd, J = 14.6, 13.0, 2.7 Hz, 1H, H-6eq), 1.39 (s, 3H, 19-CH3), 0.95 (s, 9H, H-Me3CSi), 0.17 (s, 3H, H-Me2Si), 0.14 (s, 3H, H-Me2Si). 13C NMR (151 MHz, Chloroform-d) δ 192.2 (C-2), 172.9 (C-21), 165.6 (C-16), 145.4 (C-3), 135.7 (C-4), 83.7 (C-7), 82.0 (C-13), 76.8 (C-12), 74.6 (C-20), 71.2 (C-11), 65.9 (C-15), 54.9 (17-OCH3), 53.6 (C-14), 51.0 (C-1), 45.6 (C-8), 43.2 (C-5), 43.1 (C-9), 40.7 (C-10), 29.7 (C-6), 26.4 (Me3CSi, 3C), 19.2 (Me3CSi, C), 15.8 (19-CH3), 14.7 (18-CH3), -3.4 (Me2Si), -3.6 (Me2Si), MS (ESI): m / z = 553.35 [M+H] + , 575.24 [M+Na] + , 591.22 [M+K] + Anal. calcd. for C 27 H 40 O 10 Si (552.24 g / mol).

[0049] Example 3: Preparation of compound 1

[0050] The preparation method comprises the following steps:

[0051] The hydrolysis product B3 (50 mg, 0.905 mmol) in Example 2 was dissolved in an organic solvent 0.5 mL (the organic solvent can be acetonitrile, dichloromethane, ethyl acetate, methanol), followed by the addition of 5-bromovaleryl chloride (27.1 mg, 0.1358 mmol), triethylamine (1-5 equivalents) and 4-dimethylaminopyridine (0.1-2 equivalents), wherein the solvent is dichloromethane, the addition of 3 equivalents of triethylamine and 0.5 equivalents of 4-dimethylaminopyridine is the best reaction condition. The mixture was stirred at room temperature for 8 hours. After confirming the completion of the reaction by thin layer chromatography (TLC) analysis, the reaction mixture was concentrated by vacuum evaporation. Then it was re-dissolved in tetrahydrofuran (0.5 mL), and tetrabutylammonium fluoride (57.1 mg, 0.1810 mmol) was added to break the silicon protecting group of 3-OH to generate compound 1. The crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:2) eluent to obtain compound 1. Compound 1 is a white powdery solid, with a yield of 66%.

[0052] The compound 1 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0053] 1 H NMR (600 MHz, Chloroform-d) δ 6.36 (s, 1H, 12-OH), 6.10 (s, 1H, 11-OH), 4.75 (s, 1H, H-7), 4.73 (d, J = 7.9 Hz, 1H, H-20ax), 4.23-4.26 (m, 1H, H-11), 4.19 (s, 1H, H-12), 3.84 (s, 3H, 17-OCH3), 3.79 (d, J = 7.9 Hz, 1H, H-20eq), 3.56 (t, J = 6.0 Hz, 2H, H-5'), 3.38 (s, 1H, H-14), 3.04 (d, J = 12.9 Hz, 1H, H-5), 2.96 (d, J = 15.8 Hz, 1H, H-lax), 2.45-2.40 (m, 2H, H-2'), 2.38 (m, 2H, H-leq, H-6ax), 2.15 (br s, 1H, H-9), 1.84 (d, J = 1.7 Hz, 3H, 18-CH3), 1.83-1.80 (m, 4H, H-3', H-4'), 1.73-1.80 (m, 1H, H-6eq), 1.39 (d, J = 1.1 Hz, 3H, 19-CH3). 13 C NMR (151 MHz, Chloroform-d) δ 192.2 (C-2), 172.3 (C-21), 171.5 (C-l'), 166.8 (C-16), 144.2 (C-3), 127.9 (C-4), 82.7 (C-7), 81.4 (C-13), 75.9 (C-12), 74.2 (C-20), 71.1 (C-11), 66.5 (C-15), 53.6 (17-OCH3), 53.3 (C-14), 48.7 (C-l), 45.6 (C-8), 44.6 (C-5'), 42.1 (C-5), 42.0 (C-9), 41.2 (C-10), 32.9 (C-2'), 31.6 (C-4'), 29.2 (C-6), 22.0 (C-3'), 15.6 (19-CH3), 13.5 (18-CH3), MS (ESI): m / z = 601.54 [M+H] + .Anal. calcd. for C 26 H 33 BrO 11 (600.12 g / mol).

[0054] Example 4: Preparation of compound 2

[0055] In accordance with the synthesis method of compound 1, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:2) eluent, and finally compound 2 was obtained. Compound 2 was a white powdery solid, yield: 64%.

[0056] The compound 2 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0057] 1 H NMR (600 MHz, Chloroform-d) δ 6.36 (s, 1H, OH), 6.10 (s, 1H, OH), 4.75 (s, 1H, H-7), 4.73 (d, J = 7.9 Hz, 1H, H-20ax), 4.23-4.26 (m, 1H, H-11), 4.19 (s, 1H, H-12), 3.84 (s, 3H, 17-OCH3), 3.79 (d, J = 7.9 Hz, 1H, H-20eq), 3.56 (t, J = 6.0 Hz, 2H, H-5'), 3.38 (s, 1H, H-14), 3.04 (d, J = 12.9 Hz, 1H, H-5), 2.96 (d, J = 15.8 Hz, 1H, H-1ax), 2.45-2.40 (m, 2H, H-2'), 2.38 (m, 2H, H-1eq, H-6ax), 2.15 (br s, 1H, H-9), 1.84 (d, J = 1.7 Hz, 3H, 18-CH3), 1.83-1.80 (m, 4H, H-3', H-4'), 1.73-1.80 (m, 1H, H-6eq), 1.39 (d, J = 1.1 Hz, 3H, 19-CH3). 13 C NMR (151 MHz, Chloroform-d) δ 192.3 (C-2), 172.4 (C-21), 172.2 (C-1'), 166.9 (C-16), 144.2 (C-3), 128.0 (C-4), 82.7 (C-7), 81.4 (C-13), 75.9 (C-12), 74.1 (C-20), 71.1 (C-11), 66.5 (C-15), 53.3 (17-OCH3), 53.3 (C-14), 48.7 (C-1), 45.6 (C-8), 44.6 (C-5'), 42.0 (C-5), 42.0 (C-9), 41.2 (C-10), 32.9 (C-2'), 31.6 (C-4'), 29.2 (C-6), 21.9 (C-3'), 15.6 (19-CH3), 13.5 (18-CH3), MS (ESI): m / z = 557.20 [M+H] + , 579.21 [M+Na] +Anal. calcd. for C 26 H 33 ClO 11 (556.17g / mol)。

[0058] Example 5: Preparation of compound 3

[0059] In accordance with the synthesis method of compound 1, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:2) eluent, and finally compound 3 was obtained. Compound 3 was a white powdery solid, yield: 70%.

[0060] The compound 3 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0061] 1 H NMR (600 MHz, Chloroform-d) δ 6.12 (s, 1H, 11-OH), 4.75 (s, 1H, H-7), 4.72 (d, J = 7.9 Hz, 1H, H-20ax), 4.23 (d, J = 4.37 Hz, 1H, H-11), 4.19 (s, 1H, H-12), 3.84 (s, 3H, 17-OCH3), 3.77 (d, J = 7.9 Hz, 1H, H-20eq), 3.55 (t, J = 6.0 Hz, 2H, H-4'), 3.04 (s, 1H, H-14), 2.96 (d, J = 12.9 Hz, 1H, H-5), 2.95 (d, J = 15.8 Hz, 1H, H-1ax), 2.42 (d, J = 15.8 Hz, 1H, H-leq), 2.37 (m, 1H, H-6ax), 2.15 (br s, 1H, H-9), 1.83 (d, J = 1.7 Hz, 3H, 18-CH3), 1.83-1.80 (m, 4H, H-2', H-3'), 1.75 (m, 1H, H-6eq), 1.38 (d, J = 1.1 Hz, 3H, 19-CH3). 13C NMR (151 MHz, Chloroform-d) δ 192.2 (C-2), 172.3 (C-21), 171.5 (C-l’), 166.8 (C-16), 144.2 (C-3), 127.9 (C-4), 82.7 (C-7), 81.4 (C-13), 76.0 (C-12), 74.2 (C-20), 71.1 (C-l l), 66.5 (C-15), 53.3 (17-OCH3), 53.3 (C-14), 48.7 (C-l), 45.6 (C-8), 44.6 (C-5’), 42.1 (C-5), 42.0 (C-9), 41.2 (C-10), 32.9 (C-2’), 31.6 (C-4’), 29.2 (C-6), 22.0 (C-3’), 15.6 (19-CH3), 13.5 (18-CH3), MS (ESI): m / z = 543.42 [M+H] + Anal. calcd. for C 25 H 31 ClO 11 (542.16 g / mol).

[0062] Example 6: Preparation of compound 4

[0063] In accordance with the synthesis method of compound 1, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (1:1) eluent, and finally compound 4 was obtained. Compound 4 was a white powdery solid, yield: 66%.

[0064] The compound 4 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0065] 1H NMR (600 MHz, pyridine-d5) δ 9.98 (s, 1H, 3-OH), 8.12 (d, J = 8.4 Hz, 2H, H-4', H-6'), 7.42 (d, J = 8.4 Hz, 2H, H-3', H-7'), 6.60 (s, 1H, 12-OH), 5.17-5.13 (3H, overlapped, H-7, H-15, H-20eq), 5.08 (s, 1H, H-12), 4.83 (d, J = 4.9 Hz, 1H, H-11), 4.01 (d, J = 7.9 Hz, 1H, H-20ax), 4.13-4.08 (m, 1H, H-14), 3.61 (s, 3H, 17-OCH3), 3.31 (d, J = 15.8 Hz, 1H, H-lax), 3.07 (br s, 1H, H-5), 2.57-2.51 (m, 1H, H-leq), 2.36 (dt, J = 14.3, 3.3 Hz, 1H, H-6ax), 2.69 (d, J = 4.61 Hz, 1H, H-9), 2.00 (d, J = 1.7 Hz, 3H, 18-CH3), 1.83-1.77 (m, 1H, H-6eq), 1.66 (s, 3H, 19-CH3). 13 C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 171.7 (C-21), 168.4 (C-16), 164.3 (C-l), 146.5 (C-3), 140.0 (C-5), 132.1 (C-4', C-6'), 129.5 (C-3'), 129.3 (C-7'), 128.6 (C-2'), 84.4 (C-7), 83.2 (C-13), 76.1 (C-12), 74.2 (C-20), 73.4 (C-l l), 67.5 (C-15), 52.8 (17-OCH3), 50.4 (C-l), 46.7 (C-8), 42.8 (C-5), 42.7 (C-9), 41.7 (C-10), 30.4 (C-6), 16.1 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 577.15 [M+H] + Anal. calcd. for C 28 H 29 ClO 11 (576.14 g / mol).

[0066] Example 7: Preparation of compound 5

[0067] In accordance with the synthesis method of compound 1, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (1:1) eluent, and finally compound 5 was obtained. Compound 5 was a white powdery solid, yield: 71%.

[0068] The compound 5 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0069] 1 H NMR (600 MHz, Chloroform-d) δ 7.69 (d, J = 4.1 Hz, 1H, H-5'), 6.95 (dd, J = 4.1, 1.9 Hz, H-4'), 5.43 (t, J = 1.5 Hz, 1H, H-20ax), 4.96 (d, J = 12.7 Hz, 1H, H-7), 4.84-4.80 (m, 1H, H-11), 4.77 (d, J = 7.9 Hz, 1H, H-20ax), 4.29 (d, J = 4.9 Hz, 1H, H-12), 3.79 (d, J = 7.9 Hz, 1H, H-20eq), 3.77 (s, 3H, 17-OCH3), 3.04 (d, J = 12.9 Hz, 1H, H-5), 2.95 (d, J = 15.8 Hz, 1H, H-1ax), 2.89 (d, J = 12.78 Hz, 1H, H-14), 2.30 (d, J = 15.8 Hz, 1H, H-1eq), 2.39 (d, J = 14.3 Hz, 1H, H-6ax), 2.04 (s, 1H, H-9), 1.84 (d, J = 1.7 Hz, 3H, 18-CH3), 1.80 (m, 1H, H-6eq), 1.40 (brs, 3H, 19-CH3). 13 CNMR (151 MHz, Chloroform-d) δ 191.9 (C-2), 172.3 (C-21), 169.9 (C-16), 159.8 (C-1'), 144.2 (C-3), 139.3 (C-3'), 134.9 (C-6'), 130.3 (C-5'), 127.8 (C-4'), 128.1 (C-4), 83.4 (C-7), 81.3 (C-13), 76.2 (C-12), 73.8 (C-20), 69.5 (C-11), 65.8 (C-15), 53.6 (17-OCH3), 53.4 (C-14), 48.7 (C-1), 45.1 (C-8), 43.1 (C-5), 42.3 (C-9), 41.3 (C-10), 29.2 (C-6), 15.6 (19-CH3), 13.5 (18-CH3), MS (ESI): m / z = 583.14 [M+H] +, 605.10 [M+Na] + Anal. calcd. for C 26 H 27 ClO 11 S (582.10 g / mol).

[0070] Example 8: Preparation of compound 6

[0071] In accordance with the synthesis method of compound 1, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (1:1) eluent, and finally compound 6 was obtained. Compound 6 was a white powdery solid, yield: 66%.

[0072] The compound 6 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0073] 1 H NMR (600 MHz, pyridine-d5) δ 9.17 (d, J = 2.4 Hz, 1H, H-3'), 8.29 (dd, J = 8.3, 2.4 Hz, 1H, H-7'), 7.42 (d, J = 8.3 Hz, 1H, H-6'), 5.19 (d, J = 2.8 Hz, 1H, H-15), 5.15 (d, J = 7.4 Hz, 1H, H-7), 5.07 (s, 1H, H-12), 4.84-4.81 (m, 1H, H-11), 4.11-4.07 (m, 1H, H-20eq), 4.02 (d, J = 7.9 Hz, 1H, H-20ax), 3.68 (s, 3H, 17-OCH3), 3.31 (d, J = 15.8 Hz, 1H, H-1ax), 3.07 (br s, 1H, H-5), 2.55 (d, J = 15.8 Hz, 1H, H-1eq), 2.37 (dt, J = 14.3, 3.3 Hz, 1H, H-6ax), 2.71 (d, J = 4.61 Hz, 1H, H-9), 2.00 (d, J = 1.7 Hz, 3H, 18-CH3), 1.85-1.75 (m, 1H, H-6eq), 1.66 (s, 3H, 19-CH3). 13C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 171.7 (C-21), 168.2 (C-16), 167.6 (C-1'), 156.3 (C-5'), 151.9 (C-3'), 146.4 (C-3), 140.6 (C-7'), 125.7 (C-2'), 125.1 (C-6'), 84.5 (C-7), 83.3 (C-13), 76.1 (C-12), 74.2 (C-20), 73.4 (C-11), 66.0 (C-15), 52.9 (17-OCH3), 50.4 (C-1), 46.8 (C-8), 42.8 (C-5), 42.7 (C-9), 41.7 (C-10), 30.3 (C-6), 16.1 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 578.15 [M+H] + Anal. calcd. for C 27 H 28 ClNO 11 (577.14 g / mol).

[0074] Example 9: Preparation of compound 7

[0075] In accordance with the synthesis method of compound 1, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:1) eluent, and finally compound 7 was obtained. Compound 7 was a white powdery solid, yield: 64%.

[0076] The compound 7 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0077] 1H NMR (600 MHz, pyridine-d5) δ 9.99 (s, 1H, 3-OH), 7.73 (overlapped, 2H, H-3', H-7'), 7.64-7.54 (overlapped, 2H, H-4', H-6'), 6.62 (s, 1H, 12-OH), 5.19-5.15 (2H, overlapped, H-7, H-15), 5.08 (s, 1H, H-12), 4.84 (s, 1H, H-11), 4.25-3.95 (3H, overlapped, H-14, H-20ax, H-20eq), 3.66 (s, 3H, 17-OCH3), 3.32 (d, J = 15.8 Hz, 1H, H-lax), 3.08 (br s, 1H, H-5), 2.55 (d, J = 15.8 Hz, 1H, H-leq), 2.37 (d, J = 14.3 Hz, 1H, H-6ax), 2.75-2.64 (m, 1H, H-9), 1.99 (s, 3H, 18-CH3), 1.85-1.75 (m, 1H, H-6eq), 1.67 (s, 3H, 19-CH3). 13 C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 171.7 (C-21), 168.3 (C-16), 146.4 (C-3), 134.7 (C-5'), 134.6 (C-3'), 134.1 (C-7'), 131.1 (C-2'), 126.2 (C-4'), 125.5 (C-6'), 125.1 (C-8'), 84.5 (C-7), 83.3 (C-13), 76.1 (C-12), 74.2 (C-20), 73.4 (C-11), 52.8 (17-OCH3), 50.4 (C-l), 46.8 (C-8), 42.8 (C-5), 42.8 (C-9), 41.7 (C-10), 30.4 (C-6), 16.1 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 611.18 [M+H] + ,628.20 [M+NH4] + Anal. calcd. for C 29 H 29 F3O 11 (610.17 g / mol).

[0078] Example 10: Preparation of compound 8

[0079] 3-Fluorophenylacetic acid (20.9 mg, 0.1358 mmol), l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (EDCI, 1-4 equivalents) and triethylamine (1-4 equivalents) were dissolved in an organic solvent (the organic solvent can be acetonitrile, dichloromethane, ethyl acetate, methanol) and stirred at room temperature for 2 hours. TLC analysis showed that the carboxylic acid reacted with EDCI to form an activated intermediate. Then the hydrolysis product B3 in Example 2 (50 mg, 0.0905 mmol) and 4-dimethylaminopyridine (0.1-2 equivalents) were added, and the mixture was allowed to react for 8 hours. When the organic solvent was dichloromethane, the optimal reaction conditions were 1.5 equivalents of EDCI, 2 equivalents of triethylamine, and 0.5 equivalents of 4-dimethylaminopyridine. After TLC confirmed that the reaction was complete, the reaction mixture was concentrated under reduced pressure. The product was extracted with ethyl acetate and water, and the organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure again. TBAF (57.1 mg, 0.1810 mmol) was dissolved in THF (0.5 mL) and added, and the mixture was stirred at room temperature for 0.5 hours. The target product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (2:1) as the eluent, and compound 8 was obtained as a white powdery solid with a yield of 59%.

[0080] The compound 8 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0081] 1 H NMR (600 MHz, Chloroform-d) δ 8.13 (s, 1H, H-3'), 6.11 (s, 1H, 11-OH), 4.79 (s, 1H, H-7), 4.76 (d, J = 7.9 Hz, 1H, H-20ax), 4.28 (d, J = 4.37 Hz, 1H, H-11), 4.21 (s, 1H, H-12), 3.83 (d, J = 7.9 Hz, 1H, H-20eq), 3.79 (s, 3H, 17-OCH3), 3.25 (d, J = 12.78 Hz, 1H, H-14), 2.98 (d, J = 12.9 Hz, 1H, H-5), 2.97 (d, J = 15.8 Hz, 1H, H-lax), 2.46 (d, J = 15.8 Hz, 1H, H-leq), 2.46 (d, J = 14.3 Hz, 1H, H-6ax), 2.22 (d, J = 3.90 Hz, 1H, H-9), 1.84 (d, J = 1.7 Hz, 3H, 18-CH3), 1.78 (br s, 1H, H-6eq), 1.39 (br s, 3H, 19-CH3). 13C NMR (151 MHz, Chloroform-d) δ 192.1 (C-2), 172.1 (C-21), 169.6 (C-1'), 166.6 (C-16), 162.9 (C-5', J CF = 246.3 Hz), 144.2 (C-3), 135.2 (C-3', J CF = 8.41 Hz), 130.3 (C-7', J CF = 8.41 Hz), 127.8 (C-4), 125.3 (C-8', J CF = 2.92 Hz), 116.7 (C-4', J CF = 21.15 Hz), 114.5 (C-6', J CF = 21.15 Hz), 82.7 (C-7), 81.5 (C-13), 75.9 (C-12), 74.1 (C-20), 71.1 (C-11), 65.7 (C-15), 53.6 (17-OCH3), 53.2 (C-14), 48.7 (C-1), 45.7 (C-8), 42.1 (C-5), 42.0 (C-9), 41.2 (C-10), 40.2 (C-2'), 29.8 (C-6), 15.6 (19-CH3), 14.3 (18-CH3), MS (ESI): m / z = 597.23 [M+Na] + , 613.25 [M+K] + Anal. calcd. for C 29 H 31 FO 11 (574.16 g / mol).

[0082] Example 11: Preparation of compound 9

[0083] In accordance with the synthesis method of compound 8, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (2:1) eluent, and finally compound 9 was obtained. Compound 9 was a white powdery solid, yield: 64%.

[0084] The compound 9 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0085] 1H NMR (600 MHz, pyridine-d5) δ 9.93 (s, 1H, 3-OH), 8.49 (s, 1H, H-3'), 7.62 (d, J = 3.71 Hz, 1H, H-6'), 6.58 (s, 1H, 12-OH), 5.15-4.96 (overlapped, 4H, H-7, H-12, H-15, H-20eq), 4.82 (d, J = 4.37 Hz, 1H, H-11), 4.09 (d, J = 12.56 Hz, 1H, H-14), 3.96 (d, J = 7.29 Hz, 1H, H-20ax), 3.59 (s, 3H, 17-OCH3), 3.30 (d, J = 15.8 Hz, 1H, H-lax), 3.05 (d, J = 12.90 Hz, 1H, H-5), 2.54 (d, J = 14.3 Hz, 1H, H-6ax), 2.33 (d, J = 15.8 Hz, 1H, H-leq), 2.65 (d, J = 4.61 Hz, 1H, H-9), 1.97 (d, J = 1.7 Hz, 3H, 18-CH3), 1.78 (m, 1H, H-6eq), 1.64 (s, 3H, 19-CH3). 13 C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 171.6 (C-21), 168.3 (C-16), 160.8 (C-l), 146.4 (C-3), 136.7 (C-5), 130.9 (C-6), 127.2 (C-3), 111.7 (C-4), 84.2 (C-7), 83.2 (C-13), 76.2 (C-12), 74.1 (C-20), 73.4 (C-l l), 52.7 (17-OCH3), 50.4 (C-l), 46.6 (C-8), 42.8 (C-5), 42.6 (C-9), 41.7 (C-10), 29.9 (C-6), 16.1 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 644.10 [M+NH4] + , 649.10 [M+Na] + Anal. calcd. for C 26 H 27 BrO 11 S (626.05 g / mol).

[0086] Example 12: Preparation of compound 10

[0087] In accordance with the synthesis method of compound 8, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (2:1) eluent, and finally compound 10 was obtained. Compound 10 was a white powdery solid, yield: 71%.

[0088] The compound 10 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0089] 1 H NMR (600 MHz, Chloroform-d) δ 8.13 (s, 1H, H-6'), 6.11 (s, 1H, 11-OH), 4.79 (s, 1H, H-7), 4.76 (d, J = 7.9 Hz, 1H, H-20ax), 4.28 (d, J = 4.37 Hz, 1H, H-11), 4.21 (s, 1H, H-12), 3.83 (d, J = 7.9 Hz, 1H, H-20eq), 3.79 (s, 3H, 17-OCH3), 3.25 (d, J = 12.78 Hz, 1H, H-14), 2.98 (d, J = 12.9 Hz, 1H, H-5), 2.97 (d, J = 15.8 Hz, 1H, H-1ax), 2.46 (d, J = 15.8 Hz, 1H, H-1eq), 2.46 (d, J = 14.3 Hz, 1H, H-6ax), 2.22 (d, J = 4.61 Hz, 1H, H-9), 1.84 (d, J = 1.7 Hz, 3H, 18-CH3), 1.78 (br s, 1H, H-6eq), 1.39 (br s, 3H, 19-CH3). 13 C NMR (151 MHz, Chloroform-d) δ 192.2 (C-2), 172.0 (C-21), 166.3 (C-16), 158.6 (C-1'), 152.9 (C-3'), 144.2 (C-3), 144.0 (C-5'), 131.3 (C-6'), 127.9 (C-4), 83.0 (C-7), 81.4 (C-13), 75.8 (C-12), 74.2 (C-20), 71.1 (C-11), 67.4 (C-15), 53.6 (17-OCH3), 53.5 (C-14), 48.6 (C-1), 45.6 (C-8), 42.1 (C-5), 42.0 (C-9), 41.2 (C-10), 29.8 (C-6), 15.6 (19-CH3), 14.3 (18-CH3), MS (ESI): m / z = 584.18 [M+H] + , 606.09 [M+Na] + .Anal. calcd. for C25 H 26 ClNO 11 S (583.09 g / mol).

[0090] Example 13: Preparation of compound 11

[0091] In accordance with the synthesis method of compound 8, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (4:1) eluent, and finally compound 11 was obtained. Compound 11 was a white powdery solid, yield: 55%.

[0092] Compound 11 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0093] 1 H NMR (600 MHz, pyridine-d5) δ 9.96 (s, 1H, 3-OH), 8.42 (d, J = 8.33 Hz, 1H, H-4'), 8.26 (d, J = 8.33 Hz, 1H, H-5'), 8.15 (dd, J = 7.42, 1.01 Hz, 1H, H-3'), 7.81 (dd, J = 8.33, 1.01 Hz, 1H, H-11'), 7.42 (t, J = 7.73 Hz, 1H, H-10'), 5.18-4.96 (overlapped, 4H, H-7, H-12, H-15, H-20eq), 4.85 (d, J = 4.37 Hz, 1H, H-11), 4.21 (d, J = 12.56 Hz, 1H, H-14), 4.02 (d, J = 7.29 Hz, 1H, H-20ax), 3.68 (s, 3H, 17-OCH3), 3.31 (d, J = 15.8 Hz, 1H, H-1ax), 3.07 (br s Hz 1H, H-5), 2.71 (d, J = 4.61 Hz, 1H, H-9), 2.36 (d, J = 14.3 Hz, 1H, H-6ax), 2.33 (d, J = 15.8 Hz, 1H, H-1eq), 2.00 (d, J = 1.7 Hz, 3H, 18-CH3), 1.80 (m, 1H, H-6eq), 1.67 (s, 3H, 19-CH3). 13CNMR (151 MHz, pyridine-d5) d 193.4 (C-2), 171.8 (C-21), 168.2 (C-16), 164.2 (C-1'), 149.3 (C-3'), 146.4 (C-3), 145.4 (C-7'), 138.7 (C-5'), 134.9 (C-9'), 131.3 (C-6'), 129.8 (C-11'), 128.6 (C-4), 128.5 (C-9'), 126.7 (C-4'), 122.9 (C-7'), 84.4 (C-7), 83.2 (C-13), 76.2 (C-12), 74.2 (C-20), 73.4 (C-11), 53.0 (17-OCH3), 50.4 (C-1), 46.8 (C-8), 42.8 (C-5), 42.7 (C-9), 41.7 (C-10), 29.9 (C-6), 16.1 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 672.12 [M+H] + , 689.12 [M+NH4] + , 694.19 [M+Na] + Anal. calcd. for C 31 H 30 BrNO 11 (671.10 g / mol).

[0094] Example 14: Preparation of compound 12

[0095] The preparation method comprises the following steps:

[0096] The hydrolysis product B3 (50 mg, 0.0905 mmol) in Example 2, 3-cyanopropionic acid (17.9 mg, 0.1810 mmol), N,N'-diisopropylcarbodiimide (DIC, 22.8 mg, 0.1810 mmol), 1-hydroxybenzotriazole (HOBT, 24.5 mg, 0.1810 mmol) and 4-dimethylaminopyridine (5.5 mg, 0.0453 mmol) were dissolved in dichloromethane and stirred at room temperature for 8 hours. After the reaction, the mixture was concentrated under reduced pressure. The product was extracted with ethyl acetate / 10% hydrochloric acid, ethyl acetate / saturated sodium bicarbonate and ethyl acetate / water in this order. The organic layer was dried with anhydrous sodium sulfate and then concentrated under vacuum again. TBAF (57.1 mg, 0.1810 mmol) was dissolved in THF (0.5 mL) and added, and the mixture was stirred at room temperature for 0.5 hours. After one reduction pressure concentration and extraction with ethyl acetate and water, the target product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (1:1) as an eluent to obtain compound 12. Compound 12 was a white powdery solid, and the yield was 72%.

[0097] Compound 12 obtained by column chromatography was analyzed using conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0098] 1 H NMR (600 MHz, Chloroform-d) δ 6.10 (s, 1H, 11-OH), 4.77 (s, 1H, H-7), 4.74 (d, J = 7.9 Hz, 1H, H-20ax), 4.25 (br s, 1H, H-11), 4.21 (s, 1H, H-12), 3.79 (d, J = 7.9 Hz, 1H, H-20eq), 3.81 (s, 3H, 17-OCH3), 3.05 (d, J = 12.78 Hz, 1H, H-14), 2.96 (d, J = 15.80 Hz, 1H, H-1ax), 2.94 (d, J = 12.90 Hz, 1H, H-5), 2.75 (m, 1H, H-3'), 2.68 (m, 1H, H-2'), 2.46 (d, J = 14.3 Hz, 1H, H-6ax), 2.44 (d, J = 15.80 Hz, 1H, H-1eq), 2.17 (m, 1H, H-9), 1.85 (d, J = 1.7 Hz, 3H, 18-CH3), 1.78 (m, 1H, H-6eq), 1.39 (s, 3H, 19-CH3). 13C NMR (151 MHz, Chloroform-d) δ 192.2 (C-2), 174.1 (C-21), 168.5 (C-16), 166.5 (C-1'), 144.2 (C-3), 129.0 (C-4), 118.5 (C-4'), 83.3 (C-7), 81.6 (C-13), 75.9 (C-12), 74.1 (C-20), 73.9 (C-11), 65.7 (C-15), 53.6 (17-OCH3), 53.5 (C-14), 48.6 (C-1), 45.8 (C-8), 42.2 (C-5), 42.0 (C-9), 41.2 (C-10), 32.1 (C-2'), 29.8 (C-6), 15.6 (19-CH3), 14.3 (18-CH3), 13.0 (C-3'), MS (ESI): m / z = 520.18 [M+H] + , 542.18 [M+Na] + , 537.22 [M+NH4] + Anal. calcd. for C 25 H 29 NO 11 (519.17 g / mol).

[0099] Example 15: Preparation of compound 13

[0100] In accordance with the synthesis method of compound 12, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (2:1) eluent, and finally compound 13 was obtained. Compound 13 was a white powdery solid, yield: 70%.

[0101] The compound 13 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis method, and the analysis results are as follows:

[0102] 1H NMR (600 MHz, pyridine-d5) δ 9.93 (s, 1H, 3-OH), 8.02 (d, J = 8.31 Hz, 1H, H-6'), 7.71 (br s, 1H, H-4'), 7.51 (d, J = 7.37 Hz, 1H, H-7'), 7.26 (d, J = 7.73 Hz, 1H, H-8'), δ 6.59 (s, 1H, 12-OH), 5.14-4.96 (overlapped, 4H, H-7, H-12, H-15, H-20eq), 4.78 (d, J = 4.37 Hz, 1H, H-11), 3.94 (d, J = 7.29 Hz, 1H, H-20ax), 3.83 (s, 3H, 17-OCH3), 3.75 (br s, 1H, H-2'), 3.27 (d, J = 15.8 Hz, 1H, H-lax), 3.02 (br s Hz 1H, H-5), 2.60 (d, J = 4.61 Hz, 1H, H-9), 2.50 (d, J = 14.3 Hz, 1H, H-6ax), 2.30 (d, J = 15.8 Hz, 1H, H-leq), 1.96 (d, J = 1.7 Hz, 3H, 18-CH3), 1.76 (m, 1H, H-6eq), 1.63 (s, 3H, 19-CH3). 13 C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 171.8 (C-21), 168.2 (C-16), 168.3 (C-l'), 146.4 (C-3), 134.9 (C-3'), 133.9 (C-4'), 131.3 (C-8'), 129.9 (C-6'), 139.6 (C-7'), 128.6 (C-4), 119.5 (C-9'), 113.0 (C-5'), 84.3 (C-7), 83.3 (C-13), 76.2 (C-12), 74.1 (C-20), 73.4 (C-l l), 52.8 (17-OCH3), 50.4 (C-l), 46.6 (C-8), 42.7 (C-5), 42.5 (C-9), 41.7 (C-10), 40.4 (C-2'), 29.9 (C-6), 16.0 (19-CH3), 13.8 (18-OCH3), MS (ESI): m / z = 582.08 [M+H] + , 635.29 [M+H] + Anal. calcd. for C 30 H 31 NO 11 (581.19 g / mol).

[0103] Example 16: Preparation of compound 14

[0104] In accordance with the synthesis method of compound 12, the crude product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:1) eluent, and compound 14 was finally obtained. Compound 14 was a white powdery solid, with a yield of 65%.

[0105] Compound 14 obtained by column chromatography was analyzed by using existing conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0106] 1 H NMR (600 MHz, Chloroform-d) δ 7.83 (d, J = 4.04 Hz, 1H, H-5'), 7.64 (d, J = 4.10 Hz, 1H, H-4'), 4.83 (s, 1H, H-7), 4.76 (d, J = 7.90 Hz, 1H, H-20ax), 4.29 (d, J = 4.37 Hz, 1H, H-11), 4.22 (s, 1H, H-12), 3.85 (d, J = 7.90 Hz, 1H, H-20eq), 3.73 (s, 3H, 17-OCH3), 3.20 (d, J = 12.78 Hz, 1H, H-14), 2.99 (d, J = 15.80 Hz, 1H, H-lax), 2.98 (d, J = 12.90 Hz, 1H, H-5), 2.60 (s, 3H, 8'-CH3) 2.45 (d, J = 15.80 Hz, 1H, H-leq), 2.42 (d, J = 14.3 Hz, 1H, H-6ax), 2.18 (d, J = 4.61 Hz, 1H, H-9), 1.86 (d, J = 1.7 Hz, 3H, 18-CH3), 1.78 (ddd, J = 15.40, 13.10, 2.80 Hz, 1H, H-6eq), 1.42 (s, 3H, 19-CH3). 13C NMR (151 MHz, Chloroform-d) δ 192.0 (C-2), 190.9 (C-7'), 172.0 (C-21), 166.2 (C-16), 160.2 (C-l'), 152.9 (C-5'), 150.0 (C-3'), 144.2 (C-3), 137.5 (C-6'), 135.1 (C-5), 131.9 (C-4'), 127.6 (C-4), 82.9 (C-7), 81.5 (C-13), 75.9 (C-12), 74.2 (C-20), 71.1 (C-l l), 65.3 (C-15), 53.6 (17-OCH3), 48.7 (C-l), 45.9 (C-8), 42.2 (C-5), 42.0 (C-9), 41.3 (C-10), 29.8 (C-6), 27.1 (C-8'), 15.6 (19-CH3), 13.5 (18-CH3), MS (ESI): m / z = 591.16 [M+H] + , 608.18 [M+NH4] + , 613.15 [M+Na] + Anal. calcd. for C 28 H 30 O 12 S (590.15 g / mol).

[0107] Example 17: Preparation of compound 15

[0108] The preparation method comprises the following steps:

[0109] 4-aminoindole (33.5 mg, 0.2534 mmol), carbonyldiimidazole (1-3 eq), triethylamine (0.5-2 eq) and 4-dimethylaminopyridine (0.1-2 eq) were dissolved in 1 mL of organic solvent (the organic solvent can be acetonitrile, dichloromethane, ethyl acetate, methanol) and stirred for 2 hours to form an intermediate product. Among them, the optimal reaction conditions are 1 eq of carbonyldiimidazole, 1.5 eq of triethylamine, 0.2 eq of 4-dimethylaminopyridine, and acetonitrile as the solvent. Then the hydrolysis product B3 in Example 2 (50 mg, 0.0905 mmol) was added, and the mixture was continuously stirred for 6 hours. After TLC confirmed that the reaction was completed, the reaction mixture was concentrated under reduced pressure. The product was extracted with ethyl acetate and water, and the organic phase was dried over anhydrous sodium sulfate, then concentrated under vacuum again. TBAF (57.1 mg, 0.1810 mmol) was dissolved in THF (0.5 mL) and added, and the mixture was stirred at room temperature for 0.5 hours. After one-time concentration under reduced pressure and extraction with ethyl acetate and water, the target product was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:1) as the eluent to obtain compound 15. Compound 15 is a white powdery solid, yield: 64%.

[0110] Compound 15 obtained by column chromatography was analyzed by existing conventional nuclear magnetic resonance spectrum and mass spectrometry analysis methods, and the analysis results are as follows:

[0111] 1H NMR (600 MHz, pyridine-d5) δ 12.89 (s, 1H, N-H), 9.66 (s, 1H, 3-OH), 9.02 (d, J = 1.43 Hz, H-5'), 8.43 (dd, J = 8.52, 1.43 Hz, 1H, H-7'), 7.52 (br s, 1H, H-8'), 7.41 (d, J = 8.52 Hz, 1H, H-3'), 6.57 (br s, 1H, 12-OH), 6.47 (br s, 1H, H-4'), 5.9 (s, 1H, 11-OH), 5.14-4.96 (overlapped, 5H, H-7, H-12, H-11, H-15, H-20eq), 3.94 (d, J = 7.29 Hz, 1H, H-20ax), 3.69 (s, 3H, 17-OCH3), 3.29 (d, J = 15.8 Hz, 1H, H-lax), 3.20 (br s Hz 1H, H-5), 2.75 (d, J = 4.61 Hz, 1H, H-9), 2.59 (d, J = 14.3 Hz, 1H, H-6ax), 2.30 (d, J = 15.8 Hz, 1H, H-leq), 1.97 (d, J = 1.7 Hz, 3H, 18-CH3), 1.79 (m, 1H, H-6eq), 1.70 (s, 3H, 19-CH3). 13 C NMR (151 MHz, pyridine-d5) δ 193.4 (C-2), 173.9 (C-l'), 171.4 (C-21), 167.8 (C-16), 146.3 (C-3), 140.3 (C-10'), 128.8 (C-4), 128.6 (C-9'), 127.7 (C-6'), 125.1 (C-3'), 122.0 (C-5'), 112.2 (C-7', C-8'), 103.8 (C-4'), 83.9 (C-7), 82.2 (C-13), 77.6 (C-12), 74.7 (C-20), 70.3 (C-l l), 52.8 (17-OCH3), 50.2 (C-l), 45.9 (C-8), 43.2 (C-5), 42.8 (C-9), 41.9 (C-10), 30.3 (C-6), 16.3 (19-CH3), 14.1 (18-OCH3), MS (ESI): m / z = 608.18 [M+NH4] + , 613.16 [M+Na] + Anal. calcd. for C 30 H 31 NO 11 (581.19 g / mol).

[0112] Test Example 1: Anti-attachment experiment of compounds

[0113] The attachment organism used: Balanus amphitrite cyprids;

[0114] The anti-attachment compounds used: Jacaranone derivatives, compounds 1-15;

[0115] Collection and culture of adult barnacles: Sexually mature adult barnacles were collected from the mangrove tree branches in the intertidal zone of Shanxin village, Fangchenggang city, Guangxi Zhuang Autonomous Region, China (108°11'12.32"E, 21°34'45.97"N). The culture of barnacles was carried out under laboratory conditions with the salinity of artificial seawater maintained between 27% and 28% and the temperature stabilized at approximately 28°C. Oxygenation of seawater was ensured by using an oxygen pump while simulating a natural 12-hour light cycle. Two types of feed were used for the culture of barnacles, using animal-derived Artemia nauplii and the plant-derived Phaeodactylum tricornutum as the source of nutrition. Frozen Artemia nauplii were introduced into each glass aquarium at a density of approximately 200 individuals / L. In addition, Phaeodactylum tricornutum algal liquid with an OD value of approximately 0.3 was added to the seawater containing the barnacles at a concentration of 500 mL algal liquid / 30 L seawater. The adults were given water changes and fed every three days. 600

[0116] Collection of barnacle nauplii and metamorphosis development of cyprids: During the morning hours from 9 a.m. to 12 p.m., the phototaxis of the larvae was used to guide the aggregation of barnacle nauplii by using a handheld strong light flashlight for a half hour. The aggregated nauplii were collected into a 500-milliliter beaker. Four 40-liter transparent glass aquariums were prepared, each filled with previously sterilized seawater (the salinity was strictly controlled between 29% and 30% and the temperature was controlled at 28 to 29°C). On this basis, 3 liters of rich Phaeodactylum tricornutum algal liquid were added to 24-25 liters of sterilized seawater, and 6 mL of potassium penicillin (concentration of 87.5 g / L) and 6 mL of streptomycin (50 g / L) were added to prevent bacterial contamination. A small oxygen pump was used for light aeration to ensure optimal oxygen levels. The barnacle nauplii were transferred to these aquariums for feeding, with water changes every two days and daily observations to monitor larval development. After approximately four days, some sixth-stage nauplii began to metamorphose into cyprids. These cyprids were then collected into sterile culture dishes for further observation. These culture dishes were stored in a refrigerator at a temperature of 2 to 4°C overnight, ready for subsequent studies or experiments.

[0117] Anti-attachment experiment: The compounds dissolved in DMSO were added to the 24-well plates in turn, starting from a concentration of 25 μg / mL and performing a halving dilution to test their EC 50 ​value. The volume of compound solution in each well was kept at 5 μL, with seawater as a blank control and DMSO as a negative control. Under the assistance of a stereomicroscope, 995 microliters of seawater and 21 to 25 larvae were added to each well of a 24-well plate using a 1 mL pipette. Subsequently, the 24-well plate was incubated in a dark, temperature-controlled environment for 48 hours. After incubation, the number of dead and attached Venus larvae was counted by stereomicroscope examination, and the EC that prevented the anti-attachment of barnacle Venus larvae was determined. 50 Value and LC of Venus larvae death 50 The values ​​provide an evaluation of the effectiveness of the compounds in inhibiting the attachment of barnacle larvae and their related toxicity. The results are shown in Table 1:

[0118] Table 1 Anti-adhesion activity of compounds

[0119]

[0120] In this study, the derivative compound 3 showed superior activity and lower toxicity, and its EC 50 The LC value of this compound was 0.457 μg / mL, which was significantly lower than that of the original compound brusatol (2.7826 μg / mL) and the positive control SeaNine 211 (1.9363 μg / mL). 50 / EC 50 The value was 42.2922, reflecting its extremely low toxicity. Similarly, compound 6 showed an EC 50 The value was 0.3402 μg / mL, LC 50 / EC 50 The value is 16.9633, which also indicates that it has high activity and reduced toxicity. The success of these two compounds highlights the effectiveness of structural modification by the introduction of chlorine side chains, which significantly enhances the biological activity of these compounds while effectively reducing toxicity. In contrast, although the derivative compounds 4 and 11 showed relatively high activity, their LC 50 / EC 50 The values ​​were 2.1018 and 3.4814, respectively, indicating increased toxicity, which limits their potential for practical applications. In addition, the activities of other derivatives were relatively low, but still showed certain anti-adhesion effects, although some were more toxic (see Figure 1 and Table 1 ).

[0121] Test Example 2: Resuscitation experiment of highly active compounds

[0122] In this study, eight compounds that showed significantly enhanced activity compared to brusatol (B1) (p < 0.001) were subjected to recovery experiments at a concentration of 1 μg / mL. The experiment evaluated the ability of surviving but unattached barnacle larvae to recover their attachment ability after drug treatment, and the recovery rate was defined as the proportion of recovered attachment ability. The results of the recovery experiment were compared with the LC values ​​obtained in the anti-attachment activity assay. 50 / EC 50 The toxicity predictions were highly consistent, further verifying the reliability of the experimental results.

[0123] like Figure 2 As shown, compounds 2, 3, and 6 exhibited higher recovery rates, indicating that their effects on barnacle larvae tended to be more anesthetic rather than toxic. This suggests that these compounds have lower toxicity and may be safer for non-target organisms, thus having the potential for further development and application. In contrast, compound 11 had a zero recovery rate at this concentration, indicating that its toxic effect was stronger.

[0124] The above-mentioned embodiments merely express several implementation methods of the present invention. Although the description thereof is relatively specific and detailed, it should not be understood as limiting the scope of the present invention.

Claims

1. A brucein derivative, characterized in that: It has the following structural formula: ； Wherein, R is selected from or .

2. The use of a brucein derivative according to claim 1, characterized in that: The brucein derivative is used in preventing and treating marine biofouling.

3. A method for preparing an antifouling agent for preventing and controlling marine biofouling using the brucein derivative according to claim 1.

4. The method for preparing a brucein derivative according to claim 1, wherein: The method comprises the following steps: (1) Under the catalysis of imidazole, brucein reacts with tert-butyldimethylsilyl trifluoromethanesulfonate to form 3-OH silane-protected intermediate B2; (2) Under the catalysis of NaOH, the 15-side chain of intermediate B2 is hydrolyzed to obtain the hydrolysis product B3 containing 15-OH; (3) The hydrolysis product B3 reacts with an acyl chloride reagent under the catalysis of triethylamine to obtain an intermediate product, which is then desiliconized by TBAF to obtain the final target product.

5. Use of the brucein derivative prepared according to the method of claim 4 in anti-marine biofouling.

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