Swertiol derivative and application thereof in marine organism fouling resistance

Abstract

The invention relates to the technical field of marine antifouling, in particular to a brusatol derivative and application of the brusatol derivative in marine biofouling resistance, and structurally modifies brusatol, which is one of main compounds in plant brucea javanica with wide sources, by utilizing a directional multi-target ligand strategy to obtain the brusatol derivative. Strong anti-pollution active groups (halogen, cyano groups, benzene rings, heterocyclic rings and the like) derived from the ocean are hybridized on a brusatol mother nucleus molecule, and a series of novel brusatol derivatives hybridized with the strongly anti-pollution active groups derived from the ocean are successfully synthesized. The compounds show good anti-barnacle adhesion activity in anti-adhesion experiments of barnacles, and have low toxicity to target organisms. According to the method, the variety of candidate compounds of the anti-fouling agent is enriched, a new way is provided for developing novel and efficient anti-fouling coatings, and the method has good application prospects and industrial value.

Description

[Technical field]

[0001] The invention belongs to the technical field of marine antifouling, and particularly relates to a brucein derivative and an application thereof in preventing marine biofouling. [Background technology]

[0002] Marine biofouling refers to the attachment and growth of marine organisms such as barnacles, bryozoans and tubeworms on man-made structures such as ships and offshore platforms, causing 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 has been the most economical and feasible method. However, these practices pose a great threat to the environment. The harmful chemicals released by toxic coatings will also endanger the normal growth and development of non-target organisms, and accumulate biotoxins through the food chain, ultimately endangering human health. Therefore, there is an urgent need to find new antifouling agents that are highly efficient, low-toxic and environmentally friendly to replace traditional antifouling agents. Natural products can not only effectively prevent and control biofouling organisms such as barnacles, but are also friendly and safe to the environment. They are potential natural molecular libraries for exploring new antifouling agents and are an important material basis for the subsequent independent innovation and development of new antifouling coatings.

[0003] Natural products from marine sources have long occupied an important position in antifouling research. Thanks to the rich and diverse marine organisms that can synthesize a series of natural compounds with unique structures and significant activities, many marine natural products have shown excellent activity in preventing the attachment of fouling organisms such as barnacles. However, natural products from terrestrial sources also have unique advantages, such as wide sources, convenient access, easy artificial cultivation, high yield and low production cost. A variety of natural products from terrestrial sources also show excellent antifouling activity, such as quassin, capsaicin, cypermethrin, juvenile hormone, terpenes, cyclopeptides, alkaloids, flavonoids, tannins and other phenolic compounds, coumarins, polyketides and dihydrostilbene compounds, etc., which provide an ideal choice for the sustainable development of antifouling agents. Therefore, further using the targeted multi-target ligand strategy to construct a rich variety of synergistic derivatives with strong antifouling active groups (halogen, cyano, benzene ring, heterocycle, etc.) from marine sources on the rich terrestrial antifouling active core molecules will open a new window for the exploration of new natural antifouling agents, and its application prospects are immeasurable. [Summary of the invention]

[0004] In view of the above, it is necessary to provide a terrestrial-derived bruceiol hybrid derivative and its application in anti-marine biofouling. This method enriches the types of candidate compounds for anti-fouling agents and provides a new way to develop new and efficient anti-fouling coatings, which has good application prospects and industrial value.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] A brucein derivative or a salt thereof having a hybrid marine-derived strong fouling active group, the specific structure of which is shown in the general formula (I):

[0007]

[0008] Wherein R is an antifouling active group of marine origin including halogenated alkyl, halogenated benzene ring, heterocycle and the like.

[0009] The quassinolide bruceiol derivative or a pharmaceutically acceptable salt thereof is used in marine antifouling.

[0010] The invention provides a method for preparing a novel antifouling agent for preventing and controlling marine biofouling by utilizing the brucein derivatives.

[0011] The present invention provides a method for preparing a brucein derivative, the method comprising the following steps:

[0012] (1) Under the catalysis of imidazole, brucein reacts with tert-butyldimethylsilyl trifluoromethanesulfonate to generate a 3-OH silyl protected intermediate B2;

[0013] (2) Under the catalysis of NaOH, the 15-side chain of intermediate B2 is hydrolyzed to obtain the hydrolysis product B3 containing 15-OH;

[0014] (3) The hydrolysis product B3 reacts with an acyl chloride reagent or an acid reagent under the catalysis of triethylamine to obtain an intermediate product, which is then subjected to a desilication reaction of TBAF to obtain the final target product.

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

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

[0017]

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

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

[0020] A) Bruceol (B1) reacts with tert-butyldimethylsilyl trifluoromethanesulfonate (TBSOTf) under the catalysis of imidazole to generate intermediate B2. The purpose of this step is to protect the 3-OH with good reaction activity. The reaction temperature of this step is 5-25°C and the reaction time is 12h.

[0021] B) Under the catalysis of NaOH, the 15-side chain of intermediate B2 is hydrolyzed to obtain a hydrolysis product B3 containing 15-OH; the reaction temperature of this step is 0°C and the reaction time is 8h;

[0022] C) The hydrolysis product B3 reacts with an acyl chloride reagent under the catalysis of triethylamine (Et3N) to obtain an intermediate product, and then the final target product is obtained by desiliconization reaction 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, which are recorded as compounds 1-7.

[0023] The reaction equations corresponding to the above steps are as follows:

[0024]

[0025]

[0026] The present invention provides a second class of brucein derivative compounds 8-11 and compounds 12-14, and the preparation method thereof comprises the following steps:

[0027] a) First, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) was used to activate different types of carboxylic acids to form an activated intermediate state. The reaction temperature of this step was 5-25°C and the reaction time was 1 h. The activated intermediate state was used to attack the 15-OH of the hydrolysis product B3. Under the catalysis of DMAP, the target product 8-11 was successfully obtained at the 15 position through a desiliconization reaction, which was recorded as compound 8-11. The reaction temperature of this step was 5-25°C and the reaction time was 0.5 h.

[0028] b) First, different types of carboxylic acids are activated using N,N'-diisopropylcarbodiimide (EDCI) and 1-hydroxybenzotriazole (HOBT) to form an activated intermediate state. The reaction temperature of this step is 5-25°C and the reaction time is 1 h. The activated intermediate state is used to attack the 15-OH of the hydrolysis product B3. Under the catalysis of DMAP, the target product 12-14 is successfully obtained at the 15 position through a desiliconization reaction, which is recorded as compound 12-14. The reaction temperature of this step is 5-25°C and the reaction time is 0.5 h.

[0029] The reaction equations corresponding to the above steps are 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 used to attack the 15-OH of the hydrolysis product B3, and then the target product 15 was successfully obtained at the 15 position through a desilylation reaction under the catalysis of DMAP, 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] The present invention proposes for the first time the application of brucein derivatives with hybrid marine-derived strong active groups in anti-marine biofouling. brucein is a characteristic natural quassinolide structure derived from the terrestrial plant Brucea brucea. In previous studies, the applicant found that it has significant anti-barnacle attachment activity. Based on this discovery, this study further used a targeted multi-target ligand strategy to structurally modify brucein, and constructed a rich variety of synergistic derivatives with hybrid marine-derived strong antifouling active groups to improve its antifouling performance. Based on the existing research results, the present invention significantly improves the antifouling activity of brucein 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 brucein, an effective ingredient in brucei, as a raw material, and synthesizes several brucein derivatives through an economical and feasible method. The compound has 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. LC 50 / EC 50 The value represents the toxicity level, LC 50 / EC50 Ratios greater than 15 indicated lower toxicity. p<0.05(*), p<0.01(**), p<0.001(***), and p>0.05(ns).

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

[0040] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific implementation 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-butyl dimethylsilyl 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-butyl dimethylsilyl trifluoromethanesulfonate and 8 equivalents of imidazole are the optimal reaction conditions. Subsequently, extraction was performed 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 the change in solubility, this example used ethanol and water recrystallization for the first time to purify the intermediate B2. Compared with the 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 the eluent to obtain the target product intermediate B2, which was a white powdery solid with 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(600MHz,Chloroform-d)δ6.25(s,1H,H-15),4.78(s,1H,H-7),4.72(d,J=7.9Hz,1H,H-20ax),4.26(d,J=4.2Hz,1H,H-11),4.19(s,1H,H-12),3.79(dd,J=7.9Hz,1H,H-20eq),3.78(s,3H,17-OCH3),3.13(s,1H,H-14),2.93(d,J=12.9Hz,1H,H-5),2.89(d,J=15.8Hz,1H,H-1ax),2.38(dt,J=14.7,2.9Hz,1H,H-6ax),2.33(d,J=15.8Hz,1H,H-1eq),2.18(d,J=1.3Hz,3H,5'-CH3),2.07(br s,1H,H-9),1.92(d,J=1.4Hz,3H,4'-CH3),1.85(d,J=1.7Hz,3H,18-CH3),1.75(ddd,J=14.6,13.0,2.7Hz,1H,H-6eq),1.38(d,J=1.1Hz,3H,19-CH3),0.94(s,9H,H-Me3CSi),0.17(s,3H,H-Me2Si),0.13(s,3H,H-Me2Si). 13 CNMR(151MHz,Chloroform-d)δ192.4(C-2),172.3(C-21),167.5(C-16),164.8(C-1'),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-1),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 hydrolyzate B3

[0046] The intermediate B2 (100 mg, 0.1577 mmol) was dissolved in methanol (0.5 mL), and then a base was added in a water bath at 0°C for hydrolysis, wherein 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, wherein the addition of 1.5 equivalents of sodium hydroxide is the best reaction condition. The mixture was stirred at this temperature for 8 hours, and the reaction was stopped when the reaction was complete as detected by TLC, and the remaining sodium hydroxide was neutralized to a neutral pH value with dilute hydrochloric acid. It was then extracted with ethyl acetate and water, dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude extract was purified by silica gel column chromatography using a mixture of ethyl acetate-petroleum ether (7:3) as an eluent to finally obtain the hydrolyzate B3, which was a white powdery solid with a yield of 84%.

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

[0048] 1 H NMR(600MHz,Chloroform-d)δ5.28(d,J=14.7,2.9Hz 1H,H-15),4.73(s,1H,H-7),4.71(d,J=7.9Hz,1H,H-20ax),4.24(br s,1H,H-11),4.21(s,1H,H-12),3.78(d,J=7.9Hz,1H,H-20eq),3.78(s,3H,17-OCH3),3.03(s,1H,H-14),2 .86(d,J=12.9Hz,1H,H-5),2.91(d,J=15.8Hz,1H,H-1ax),2.38(dt,J=14.7,2.9Hz,1H,H-6ax),2.28(d,J=1 5.8Hz,1H,H-1eq),2.02(d,J=4.61Hz,1H,H-9),1.85(d,J=1.7Hz,3H,18-CH3),1.77(ddd,J=14.6,13.0,2. 7Hz,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(151MHz,Chloroform-d)δ192.2(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 hydrolyzate B3 (50 mg, 0905 mmol) in Example 2 was dissolved in 0.5 mL of an organic solvent (the organic solvent may 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 was dichloromethane, 3 equivalents of triethylamine, and 0.5 equivalents of 4-dimethylaminopyridine were added for the optimal reaction conditions. The mixture was stirred at room temperature for 8 hours. After the reaction was confirmed to be complete by thin layer chromatography (TLC) analysis, the reaction mixture was concentrated by vacuum evaporation. It was then redissolved in tetrahydrofuran (0.5 mL), tetrabutylammonium fluoride (57.1 mg, 0.1810 mmol) was added, and the reaction was allowed to proceed for about 30 minutes to disconnect the silicon protecting group of 3-OH to generate compound 1. The crude extract 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 using conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0053] 1 H NMR(600MHz,Chloroform-d)δ6.36(s,1H,12-OH),6.10(s,1H,11-OH),4.75(s,1H,H-7),4.73(d ,J=7.9Hz,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=7.9Hz,1H,H-20eq),3.56(t,J=6.0Hz,2H,H-5'),3.38(s,1H,H-14),3.04(d,J=12.9Hz,1H,H -5),2.96(d,J=15.8Hz,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=1.7Hz,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=1.1Hz,3H,19-CH3). 13 C NMR(151MHz,Chloroform-d)δ192.2(C-2),172.3(C-21),171.5(C-1'),166.8(C-16),144.2(C-3),1 27.9(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-OC H3),53.3(C-14),48.7(C-1),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 B O 11 (600.12 g / mol).

[0054] Example 4: Preparation of Compound 2

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

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

[0057] 1 H NMR(600MHz,Chloroform-d)δ6.36(s,1H,OH),6.10(s,1H,OH),4.75(s,1H,H-7),4.73(d,J=7. 9Hz,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 =7.9Hz,1H,H-20eq),3.56(t,J=6.0Hz,2H,H-5'),3.38(s,1H,H-14),3.04(d,J=12.9Hz,1H,H- 5),2.96(d,J=15.8Hz,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=1.7Hz,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=1.1Hz,3H,19-CH3). 13 C NMR(151MHz,Chloroform-d)δ192.3(C-2),172.4(C-21),172.2(C-1'),166.9(C-16),144.2(C-3),1 28.0(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-OC H3),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 C1O 11 (556.17 g / mol).

[0058] Example 5: Preparation of Compound 3

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

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

[0061] 1 H NMR(600MHz,Chloroform-d)δ6.12(s,1H,11-OH),4.75(s,1H,H-7),4.72(d,J=7.9Hz,1H,H- 20ax),4.23(d,J=4.37Hz,1H,H-11),4.19(s,1H,H-12),3.84(s,3H,17-OCH3),3.77(d,J=7. 9Hz,1H,H-20eq),3.55(t,J=6.0Hz,2H,H-4'),3.04(s,1H,H-14),2.96(d,J=12.9Hz,1H,H-5 ),2.95(d,J=15.8Hz,1H,H-1ax),2.42(d,J=15.8Hz,1H,H-1eq),2.37(m,1H,H-6ax),2.15(br s,1H,H-9),1.83(d,J=1.7Hz,3H,18-CH3),1.83-1.80(m,4H,H-2',H-3'),1.75(m,1H,H-6eq),1.38(d,J=1.1Hz,3H,19-CH3). 13C NMR(151MHz,Chloroform-d)δ192.2(C-2),172.3(C-21),171.5(C-1'),166.8(C-16),144.2(C-3),1 27.9(C-4),82.7(C-7),81.4(C-13),76.0(C-12),74.2(C-20),71.1(C-11),66.5(C-15),53.3(17-OC H3),53.3(C-14),48.7(C-1),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 C1O 11 (542.16 g / mol).

[0062] Example 6: Preparation of Compound 4

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

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

[0065] 1H NMR (600MHz, pyridine-d5) δ9.98 (s, 1H, 3-OH), 8.12 (d, J = 8.4Hz, 2H, H-4', H-6'), 7.42 (d,J=8.4Hz,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=4.9Hz,1H,H-11),4.01(d,J=7.9Hz,1H,H-20ax) ,4.13-4.08(m,1H,H-14),3.61(s,3H,17-OCH3),3.31(d,J=15.8Hz,1H,H-1ax),3.07(br s,1H,H-5),2.57-2.51(m,1H,H-1eq),2.36(dt,J=14.3,3.3Hz,1H,H-6ax),2.69(d,J=4.61 Hz,1H,H-9),2.00(d,J=1.7Hz,3H,18-CH3),1.83-1.77(m,1H,H-6eq),1.66(s,3H,19-CH3). 13 C NMR(151MHz,pyridine-d5)δ193.4(C-2),171.7(C-21),168.4(C-16),164.3(C-1'),146.5(C-3),14 0.0(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-11),67.5(C-15),52.8(17-OCH3),50.4(C-1),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 C1O 11 (576.14 g / mol).

[0066] Example 7: Preparation of Compound 5

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

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

[0069] 1 H NMR(600MHz,Chloroform-d)δ7.69(d,J=4.1Hz,1H,H-5'),6.95(dd,J=4.1,1.9Hz,H-4'),5.43(t,J=1.5Hz,1H,H-20ax),4.96(d,J=1 2.7Hz,1H,H-7),4.84-4.80(m,1H,H-11),4.77(d,J=7.9Hz,1H,H-20ax),4.29(d,J=4.9Hz,1H,H-12),3.79(d,J=7.9Hz,1H,H-20eq),3 .77(s,3H,17-OCH3),3.04(d,J=12.9Hz,1H,H-5),2.95(d,J=15.8Hz,1H,H-1ax),2.89(d,J=12.78Hz,1H,H-14),2.30(d,J=15.8Hz,1H ,H-1eq),2.39(d,J=14.3Hz,1H,H-6ax),2.04(s,1H,H-9),1.84(d,J=1.7Hz,3H,18-CH3),1.80(m,1H,H-6eq),1.40(brs,3H,19-CH3). 13 CNMR(151MHz,Chloroform-d)δ191.9(C-2),172.3(C-21),169.9(C-16),159.8(C-1'),144.2(C-3),1 39.3(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 C1O 11 S (582.10 g / mol).

[0070] Example 8: Preparation of Compound 6

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

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

[0073] 1 H NMR(600MHz,pyridine-d5)δ9.17(d,J=2.4Hz,1H,H-3'),8.29(dd,J=8.3,2.4Hz,1H, H-7'),7.42(d,J=8.3Hz,1H,H-6'),5.19(d,J=2.8Hz,1H,H-15),5.15(d,J=7.4Hz,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=7.9Hz,1H,H-20ax),3.68(s,3H,17-OCH3),3.31(d,J=15.8Hz,1H,H-1ax),3.07(br s,1H,H-5),2.55(d,J=15.8Hz,1H,H-1eq),2.37(dt,J=14.3,3.3Hz,1H,H-6ax),2.71(d,J=4. 61Hz,1H,H-9),2.00(d,J=1.7Hz,3H,18-CH3),1.85-1.75(m,1H,H-6eq),1.66(s,3H,19-CH3). 13C NMR(151MHz,pyridine-d5)δ193.4(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] Consistent with the synthesis method of compound 1, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (3:1) as eluent to finally obtain compound 7. Compound 7 was a white powdery solid with a yield of 64%.

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

[0077] 1H NMR(600MHz,pyridine-d5)δ9.99(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,overlap ped,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=15.8Hz,1H,H-1ax),3.08(br s,1H,H-5),2.55(d,J=15.8Hz,1H,H-1eq),2.37(d,J=14.3Hz,1H,H-6ax),2.75-2.6 4(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(151MHz,pyridine-d5)δ193.4(C-2),171.7(C-21),168.3(C-16),146.4(C-3),134.7(C-5'),1 34.6(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-1),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), 1-(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 hydrolyzate B3 (50 mg, 0.0905 mmol) and 4-dimethylaminopyridine (0.1-2 equivalents) in Example 2 were added, and the mixture continued to react for 8 hours. The organic solvent was dichloromethane, and the optimal reaction conditions were when EDCI 1.5 equivalents, triethylamine 2 equivalents, and 4-dimethylaminopyridine 0.5 equivalents were added. 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 concentrated again under reduced pressure. 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 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 (2:1) as eluent to obtain compound 8, and compound 7 was a white powdery solid, with a yield of 59%.

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

[0081] 1 H NMR(600MHz,Chloroform-d)δ8.13(s,1H,H-3'),6.11(s,1H,11-OH),4.79(s,1H,H-7),4.76(d,J=7.9Hz,1H,H -20ax),4.28(d,J=4.37Hz,1H,H-11),4.21(s,1H,H-12),3.83(d,J=7.9Hz,1H,H-20eq),3.79(s,3H,17-OCH3), 3.25(d,J=12.78Hz,1H,H-14),2.98(d,J=12.9Hz,1H,H-5),2.97(d,J=15.8Hz,1H,H-1ax),2.46(d,J=15.8Hz, 1H,H-1eq),2.46(d,J=14.3Hz,1H,H-6ax),2.22(d,J=3.90Hz,1H,H-9),1.84(d,J=1.7Hz,3H,18-CH3),1.78(br s,1H,H-6eq),1.39(br s,3H,19-CH3). 13C NMR(151MHz,Chloroform-d)δ192.1(C-2),172.1(C-21),169.6(C-1'),166.6(C-16),162.9(C-5',J CF =246.3Hz),144.2(C-3),135.2(C-3',J CF =8.41Hz),130.3(C-7',J CF =8.41Hz),127.8(C-4),125.3(C-8',J CF =2.92Hz),116.7(C-4',J CF =21.15Hz),114.5(C-6',J CF =21.15Hz),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),4 5.7(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.forC 29 H 31 FO 11 (574.16 g / mol).

[0082] Example 11: Preparation of Compound 9

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

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

[0085] 1H NMR(600MHz,pyridine-d5)δ9.93(s,1H,3-OH),8.49(s,1H,H-3'),7.62(d,J=3.71Hz,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=4.37Hz,1H,H-11),4.09(d,J=12.56Hz,1H,H-14),3.96(d,J=7.29Hz,1H, H-20ax),3.59(s,3H,17-OCH3),3.30(d,J=15.8Hz,1H,H-1ax),3.05(d,J=12.90Hz,1H,H-5),2.54(d,J=14.3Hz,1H,H-6ax),2.3 3(d,J=15.8Hz,1H,H-1eq),2.65(d,J=4.61Hz,1H,H-9),1.97(d,J=1.7Hz,3H,18-CH3),1.78(m,1H,H-6eq),1.64(s,3H,19-CH3). 13 C NMR(151MHz,pyridine-d5)δ193.4(C-2),171.6(C-21),168.3(C-16),160.8(C-1'),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-11),52.7(17-OCH3),50.4(C-1),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 B O 11 S (626.05 g / mol).

[0086] Example 12: Preparation of Compound 10

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

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

[0089] 1 H NMR(600MHz,Chloroform-d)δ8.13(s,1H,H-6'),6.11(s,1H,11-OH),4.79(s,1H,H-7),4.76(d,J=7.9Hz,1H,H -20ax),4.28(d,J=4.37Hz,1H,H-11),4.21(s,1H,H-12),3.83(d,J=7.9Hz,1H,H-20eq),3.79(s,3H,17-OCH3), 3.25(d,J=12.78Hz,1H,H-14),2.98(d,J=12.9Hz,1H,H-5),2.97(d,J=15.8Hz,1H,H-1ax),2.46(d,J=15.8Hz, 1H,H-1eq),2.46(d,J=14.3Hz,1H,H-6ax),2.22(d,J=4.61Hz,1H,H-9),1.84(d,J=1.7Hz,3H,18-CH3),1.78(br s,1H,H-6eq),1.39(br s,3H,19-CH3). 13 C NMR(151MHz,Chloroform-d)δ192.2(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] Consistent with the synthesis method of compound 8, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (4:1) as eluent to finally obtain compound 11. Compound 11 was a white powdery solid with a yield of 55%.

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

[0093] 1 H NMR(600MHz,pyridine-d5)δ9.96(s,1H,3-OH),8.42(d,J=8.33Hz,1H,H-4'),8.26(d,J=8.33Hz,1H,H-5' ),8.15(dd,J=7.42,1.01Hz,1H,H-3'),7.81(dd,J=8.33,1.01Hz,1H,H-11'),7.42(t,J=7.73Hz,1H,H-10' ),5.18-4.96(overlapped,4H,H-7,H-12,H-15,H-20eq),4.85(d,J=4.37Hz,1H,H-11),4.21(d,J=12.56Hz ,1H,H-14),4.02(d,J=7.29Hz,1H,H-20ax),3.68(s,3H,17-OCH3),3.31(d,J=15.8Hz,1H,H-1ax),3.07(br s Hz1H,H-5),2.71(d,J=4.61Hz,1H,H-9),2.36(d,J=14.3Hz,1H,H-6ax),2.33(d,J=15.8 Hz,1H,H-1eq),2.00(d,J=1.7Hz,3H,18-CH3),1.80(m,1H,H-6eq),1.67(s,3H,19-CH3). 13CNMR(151MHz,pyridine-d5)δ193.4(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 hydrolyzate B3 (50 mg, 0.0905 mmol), 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) in Example 2 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 turn. The organic layer was dried over anhydrous sodium sulfate and concentrated again under vacuum. 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 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 (1:1) as eluent to obtain compound 12. Compound 12 was a white powdery solid with a yield of 72%.

[0097] The 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(600MHz,Chloroform-d)δ6.10(s,1H,11-OH),4.77(s,1H,H-7),4.74(d,J=7.9Hz,1H,H-20ax),4.25(br s,1H,H-11),4.21(s,1H,H-12),3.79(d,J=7.9Hz,1H,H-20eq),3.81(s,3H,17-OCH3),3.05( d,J=12.78Hz,1H,H-14),2.96(d,J=15.80Hz,1H,H-1ax),2.94(d,J=12.90Hz,1H,H-5),2.75( m,1H,H-3'),2.68(m,1H,H-2'),2.46(d,J=14.3Hz,1H,H-6ax),2.44(d,J=15.80Hz,1H,H-1eq ),2.17(m,1H,H-9),1.85(d,J=1.7Hz,3H,18-CH3),1.78(m,1H,H-6eq),1.39(s,3H,19-CH3). 13C NMR(151MHz,Chloroform-d)δ192.2(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] Consistent with the synthesis method of compound 12, the crude extract was purified by silica gel column chromatography using ethyl acetate-petroleum ether (2:1) as eluent to finally obtain compound 13. Compound 13 was a white powdery solid with a yield of 70%.

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

[0102] 1H NMR(600MHz,pyridine-d5)δ9.93(s,1H,3-OH),8.02(d,J=8.31Hz,1H,H-6'),7.71(br s,1H,H-4'),7.51(d,J=7.37Hz,1H,H-7'),7.26(d,J=7.73Hz,1H,H-8'),δ6.59(s,1H,12-OH),5.14-4.96(overlapped,4H,H-7,H-12,H-15,H-20eq),4.78(d,J=4.37Hz,1H,H-11),3.94(d,J=7.29Hz,1H,H-20ax),3.83(s,3H,17-OCH3),3.75(br s,1H,H-2'),3.27(d,J=15.8Hz,1H,H-1ax),3.02(br s Hz 1H,H-5),2.60(d,J=4.61Hz,1H,H-9),2.50(d,J=14.3Hz,1H,H-6ax),2.30(d,J=15.8Hz,1H,H-1eq),1.96(d,J=1.7Hz,3H,18-CH3),1.76(m,1H,H-6eq),1.63(s,3H,19-CH3). 13 C NMR(151MHz,pyridine-d5)δ193.4(C-2),171.8(C-21),168.2(C-16),168.3(C-1'),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-11),52.8(17-OCH3),50.4(C-1),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 (581.19g / mol)。

[0103] Example 16: Preparation of Compound 14

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

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

[0106] 1 H NMR(600MHz,Chloroform-d)δ7.83(d,J=4.04Hz,1H,H-5'),7.64(d,J=4.10Hz,1H,H-4'),4.83(s,1H,H-7),4.76(d,J=7.90Hz,1H,H-20ax ),4.29(d,J=4.37Hz,1H,H-11),4.22(s,1H,H-12),3.85(d,J=7.90Hz,1H,H-20eq),3.73(s,3H,17-OCH3),3.20(d,J=12.78Hz,1H,H-14), 2.99(d,J=15.80Hz,1H,H-1ax),2.98(d,J=12.90Hz,1H,H-5),2.60(s,3H,8'-CH3)2.45(d,J=15.80Hz,1H,H-1eq),2.42(d,J=14.3Hz,1H, H-6ax), 2.18 (d, J = 4.61Hz, 1H, H-9), 1.86 (d, J = 1.7Hz, 3H, 18-CH3), 1.78 (ddd, J = 15.40, 13.10, 2.80Hz, 1H, H-6eq), 1.42 (s, 3H, 19-CH3). 13C NMR(151MHz,Chloroform-d)δ192.0(C-2),190.9(C-7'),172.0(C-21),166.2(C-16),160.2(C-1'),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-11),65.3(C-15),53.6(17-OCH3),48.7(C-1),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 (33.5mg, 0.2534mmol), carbonyl diimidazole (1-3 equivalents), triethylamine (0.5-2 equivalents) and 4-dimethylaminopyridine (0.1-2 equivalents) were dissolved in 1mL of an 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 addition of 1 equivalent of carbonyl diimidazole, 1.5 equivalents of triethylamine, 0.2 equivalents of 4-dimethylaminopyridine, and the solvent being acetonitrile is the best reaction condition. Then the hydrolyzate B3 (50mg, 0.0905mmol) in Example 2 was added, and the mixture was stirred for 6 hours. 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 phase was dried over anhydrous sodium sulfate and concentrated again under vacuum. TBAF (57.1mg, 0.1810mmol) was dissolved in THF (0.5mL) and added, and the mixture was stirred at room temperature for 0.5 hours. After 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 eluent to obtain compound 15. Compound 15 was a white powdery solid with a yield of 64%.

[0110] The compound 15 obtained by column chromatography was analyzed using conventional nuclear magnetic resonance spectroscopy and mass spectrometry analysis methods, and the analysis results are as follows:

[0111] 1H NMR(600MHz,pyridine-d5)δ12.89(s,1H,N-H),9.66(s,1H,3-OH),9.02(d,J=1.43Hz,H-5'),8.43(dd,J=8.52,1.43Hz,1H,H-7'),7.52(br s,1H,H-8'),7.41(d,J=8.52Hz,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=7.29Hz,1H,H-20ax),3.69(s,3H,17-OCH3),3.29(d,J=15.8Hz,1H,H-1ax),3.20(br s Hz 1H,H-5),2.75(d,J=4.61Hz,1H,H-9),2.59(d,J=14.3Hz,1H,H-6ax),2.30(d,J=15.8Hz,1H,H-1eq),1.97(d,J=1.7Hz,3H,18-CH3),1.79(m,1H,H-6eq),1.70(s,3H,19-CH3). 13 C NMR(151MHz,pyridine-d5)δ193.4(C-2),173.9(C-1'),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-11),52.8(17-OCH3),50.2(C-1),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.forC 30 H 31 NO 11 (581.19g / mol)。

[0112] Test Example 1: Compound Anti-Adhesion Test

[0113] Used biofouling organisms: Balanus amphitrite larvae;

[0114] Anti-adhesion compounds used: brucein derivatives, compounds 1-15;

[0115] Collection and culture of adult barnacles: Sexually mature adult barnacles were collected from mangrove branches in the coastal mudflat area of ​​Shanxin Village, Fangchenggang City, Guangxi Zhuang Autonomous Region, China (108°11'12.32"E, 21°34'45.97"N). The culture process of barnacles was carried out under laboratory conditions, and the salinity of artificial seawater was maintained between 27% and 28%, and the temperature was stabilized at approximately 28°C. The oxygenation of the 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 plant-derived Brown Finger Algae as nutrient sources. Freeze-dried Artemia nauplii were placed in each glass tank at a density of approximately 200 / L. In addition, OD was added to the seawater containing barnacles. 600 The concentration of the algae solution of the triangular brown finger algae with a value of about 0.3 was 500mL of the algae solution / 30L of seawater. The water was changed and the adults were fed every three days.

[0116] Collection of barnacle nauplii and metamorphosis culture of Venus larvae: Between 9 am and 12 pm, the larvae were guided to aggregate by the phototaxis of the larvae by using a handheld strong flashlight for half an hour. The aggregated nauplii were collected in a 500 ml beaker. Four 40-liter transparent glass tanks were prepared, each filled with previously sterilized seawater (salinity was strictly controlled between 29% and 30%, and temperature was controlled between 28 and 29°C). On this basis, 3 liters of rich triangular brown finger algae solution were added to 24-25 liters of sterilized seawater, and 6 mL of penicillin potassium (concentration of 87.5 g / L) and 6 mL of streptomycin (50 g / L) were added to prevent bacterial contamination. Light oxygenation was performed using a small oxygen pump to ensure optimal oxygen levels. Barnacle nauplii were transferred to these tanks for rearing, the water was changed every two days, and daily observations were made to monitor larval development. After about four days, some of the sixth-stage nauplii begin metamorphosis into Venus larvae. These Venus larvae are then collected into sterile petri dishes for further observation. These petri dishes are kept in a refrigerator, maintained at 2 to 4°C, overnight, ready for subsequent studies or experiments.

[0117] Anti-adhesion assay: Compounds dissolved in DMSO were sequentially added to a 24-well plate, starting at a concentration of 25 μg / mL and diluted in half to test their EC 50value. 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 to prevent 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, derivative compound 3 showed superior activity and lower toxicity, with an EC 50 The value was 0.457 μg / mL, which was significantly lower than 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 high activity and reduced toxicity. The success of these two compounds highlights the effectiveness of structural modification through the introduction of chlorine side chains, which significantly enhances the biological activity of these compounds while effectively reducing toxicity. In contrast, although 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, thus limiting its potential for practical application. 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] Eight compounds with significantly enhanced activity compared to brusatol (B1) (p < 0.001) were subjected to resuscitation experiments at a concentration of 1 μg / mL. The experiment evaluated the recovery of the attachment ability of surviving but unattached barnacle larvae (cyprids) after drug treatment, and the proportion of recovered attachment ability was defined as the recovery rate. The results of the resuscitation experiment were compared with the LC 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 showed higher recovery rates, indicating that their effects on barnacle larvae tend to be more anesthetic rather than toxic, which indicates that these compounds have lower toxicity and may be safer for non-target organisms, and therefore have 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 modes of the present invention, and the description thereof is relatively specific and detailed, but 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: In the formula, R is an antifouling active group containing marine origin.

2. A brucein derivative according to claim 1, characterized in that: The antifouling active groups containing marine sources include halogenated alkyl groups, halogenated benzene rings, and heterocycles.

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

4. A method for preparing a new antifouling agent for preventing and controlling marine biofouling using the brucein derivatives described in claim 1.

5. The method for preparing a brucein derivative according to claim 1, characterized in that: The method comprises the following steps: (1) Under the catalysis of imidazole, brucein reacts with tert-butyldimethylsilyl trifluoromethanesulfonate to generate 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 or an acid reagent under the catalysis of triethylamine to obtain an intermediate product, which is then subjected to a desilication reaction of TBAF to obtain the final target product.

6. The method for preparing a brucein derivative according to claim 1, characterized in that: The method comprises the following steps: (1) Under the catalysis of imidazole, brucein reacts with tert-butyldimethylsilyl trifluoromethanesulfonate to generate 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 hydrolyzate B3 is dissolved in dichloromethane, followed by the addition of 5-bromovaleryl chloride, triethylamine and 4-dimethylaminopyridine, and the resulting mixture is stirred at room temperature for 8 hours. After the completion of the reaction is confirmed by thin layer chromatography analysis, the reaction mixture is concentrated by vacuum evaporation, and then redissolved in tetrahydrofuran, and tetrabutylammonium fluoride is added to react to disconnect the silane protecting group of 3-OH. The crude extract is purified by silica gel column chromatography using ethyl acetate-petroleum ether as an eluent to finally obtain the brucein derivative.

7. The method according to claim 6, characterized in that The brucein derivative has the following nuclear magnetic resonance spectrum and mass spectrum data: 1 H NMR(600MHz,Chloroform-d)δ6.12(s,1H,11-OH),4.75(s,1H,H-7),4.72(d,J=7.9Hz,1H,H-20ax),4.23(d,J=4.37Hz,1H,H-11),4.19(s,1H,H-12),3.84(s,3H,17-OCH3),3.77(d,J=7.9Hz,1H,H-20eq),3.55(t,J=6.0Hz,2H,H-4'),3.04(s,1H,H-14),2.96(d,J=12.9Hz,1H,H-5),2.95(d,J=15.8Hz,1H,H-1ax),2.42(d,J=15.8Hz,1H,H-1eq),2.37(m,1H,H-6ax),2.15(br s,1H,H-9),1.83(d,J=1.7Hz,3H,18-CH3),1.83-1.80(m,4H,H-2',H-3'),1.75(m,1H,H-6eq),1.38(d,J=1.1Hz,3H,19-CH3). 13 C NMR(151MHz,Chloroform-d)δ192.2(C-2),172.3(C-21),171.5(C-1'),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-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'),42.1(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 (542.16g / mol)。 8. Use of the brucein derivative prepared according to the method according to any one of claims 4 to 7 in anti-marine biofouling.

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