A heteroterpene derivative of natural product oxirapentyn B, and preparation method and application thereof

Abstract

The present application belongs to the technical field of biological medicine, and particularly relates to a heteroterpene derivative of natural product Oxirapentyn B, a preparation method and application thereof. The heteroterpene derivative shows significant anti-inflammatory activity, can effectively inhibit the production of NO of inflammatory cells, and has low cytotoxicity and good safety, and is very suitable for being prepared into anti-inflammatory drugs. Moreover, the heteroterpene derivative takes the marine fungus-derived heteroterpene Oxirapentyn B as raw material, has the characteristics of microbial scale fermentation, simple production process, short cycle and low product cost, and has wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine. More specifically, it relates to a meroterpenoid derivative of natural product Oxirapentyn B, a preparation method and application thereof. BACKGROUND

[0002] Marine natural products have the characteristics of novel structure and significant activity, among which marine microbial natural products are the research hotspot in the field of anti-inflammatory drugs. For example, Zhao Huan et al. (Zhao H, Zou J, Xu W, et al. Diisoprenyl-cyclohexene / ane-type meroterpenoids from Biscogniauxia sp. and their anti-inflammatory activities [J]. J. Org. Chem. 2021, 86, 16, 11177-11188. DOI: 10.1021 / acs.joc.1c00369.). Our research found that the natural product Oxirapentyn B and its structural analogs isolated from the fermentation product of the sea squirt-derived fungus Beauveria felina SYSU-MS7908 have certain anti-inflammatory activity, but the anti-inflammatory activity of the natural product Oxirapentyn B is not high, and the anti-inflammatory effect is lower than that of the positive control indomethacin. In order to improve the application value of the natural product Oxirapentyn B, it is urgent to improve its structure and improve its anti-inflammatory activity.

[0003] SUMMARY

[0004] The technical problem to be solved by the present application is to overcome the defects and deficiencies of the existing natural product Oxirapentyn B with low anti-inflammatory activity, and to provide a meroterpenoid derivative of natural product Oxirapentyn B with better anti-inflammatory activity and good safety.

[0005] The purpose of the present application is to provide a preparation method of the meroterpenoid derivative of natural product Oxirapentyn B.

[0006] Another purpose of the present application is to provide the application of the meroterpenoid derivative of natural product Oxirapentyn B in preparing anti-inflammatory drugs.

[0007] The above purposes of the present application are achieved by the following technical solutions:

[0008] A meroterpenoid derivative of natural product Oxirapentyn B has any one of the following structures:

[0009]

[0010] wherein R1, R2, R3 are each independently selected from C 1～15 alkyl or substituted C 1～15 alkyl, C 2～5 alkenyl or substituted C 2～5 alkenyl, phenyl or substituted phenyl, naphthyl or substituted naphthyl, quinolinyl, tetrahydronaphthyl, cyclohexylphenyl, biphenyl;

[0011] said substituents are each independently selected from phenyl, carboxyl, C 1～5 alkyl, halogen, nitro, cyano, hydroxyl, C 1～5 alkyl substituted amine, C 1～5 alkoxy, halogenated C 1～5 alkyl, amido.

[0012] Preferably, said R1 is selected from C 1～15 alkyl or substituted C 1～15 alkyl, C 2～5 alkenyl or substituted C 2～5 alkenyl, phenyl or substituted phenyl;

[0013] said R2 is selected from C 1～5 alkyl, phenyl or substituted phenyl, naphthyl or substituted naphthyl, quinolinyl, tetrahydronaphthyl, cyclohexylphenyl, biphenyl;

[0014] said R3 is selected from C 1～5 alkyl, benzyl, phenyl;

[0015] said substituents are each independently selected from phenyl, carboxyl, C 1～5 alkyl, halogen, nitro, cyano, hydroxyl, methyl substituted amine, C 1～5 alkoxy, halogenated C 1～5 alkyl, amido.

[0016] More preferably, said R1 is selected from C 1～15 alkyl or substituted C 1～5 alkyl, methyl substituted ethenyl, phenyl or substituted phenyl;

[0017] said R2 is selected from C 1～5 alkyl, phenyl or substituted phenyl, naphthyl or substituted naphthyl, quinolinyl, tetrahydronaphthyl, cyclohexylphenyl, biphenyl;

[0018] said R3 is selected from C 1～5 alkyl, benzyl, phenyl;

[0019] each of said substituted substituents is independently selected from the group consisting of phenyl, carboxyl, methyl, halogen, nitro, cyano, hydroxyl, methyl substituted amine, methoxy, trifluoromethyl, acetylamide.

[0020] In particular, said R1 is selected from any one of the following structures: -CH3, -CH2CH3, -(CH2)3CH3, -(CH2)5CH3, -(CH2) 12 CH3, -(CH2)3Ph, -(CH2)2COOH, -Ph,

[0021] Said R2 is selected from any one of the following structures: -CH3, -CH2CH3, -PhMe, -Ph,

[0022] Said R3 is selected from any one of the following structures: -CH3, -(CH2)2CH3, -(CH2)4CH3, -CH2Ph, -Ph.

[0023] In addition, the present application also provides a preparation method of the heteroterpene derivative of the natural product Oxirapentyn B, method one specifically comprising the following steps:

[0024] Oxirapentyn B is reacted with an acyl chloride reagent at room temperature in the presence of an organic solvent, a catalyst and a basic reagent until the reaction is complete (preferably the reaction time is 1-5 h), and then the product is obtained after post-processing; the acyl chloride reagent is ClCO-R1, ClSO2-R2 or ClCO2-R3, wherein the definitions of R1, R2 and R3 are consistent with any of the above.

[0025] In addition, the present application also provides a preparation method of the heteroterpene derivative of the natural product Oxirapentyn B, method two specifically comprising the following steps:

[0026] Oxirapentyn B is reacted with an anhydride reagent at room temperature in the presence of an organic solvent and a catalyst until the reaction is complete (preferably the reaction time is 1-5 h), and then the product is obtained after post-processing; the anhydride reagent is O(CO-R1)2, O(SO2-R2)2 or O(CO2-R3)2, wherein the definitions of R1, R2 and R3 are consistent with any of the above.

[0027] The present application also provides a preparation method of the heteroterpene derivative of the natural product Oxirapentyn B, method three specifically comprising the following steps:

[0028] Oxirapentyn B is reacted with an organic acid reagent R1COOH, R2SO3H or R3CO3H (wherein R1, R2, R3 are defined as any of the above) in the presence of an organic solvent, a catalyst and a condensing agent at room temperature until the reaction is completed (preferably for 1-5 hours), and then the product is obtained by post-treatment.

[0029] Further, in the methods one to three, the catalyst is an organic base selected from one or more of 4-dimethylaminopyridine (DMAP), pyridine, triethylamine, diethylamine, ethylenediamine, N,N-diisopropylethylamine.

[0030] Still further, in the methods one to three, the organic solvent is selected from one or more of dichloromethane, tetrahydrofuran, N,N-dimethylformamide (DMF), 1,4-dioxane, acetonitrile, toluene, chloroform, triethylamine, acetone, pyridine; preferably, dichloromethane.

[0031] Preferably, in the methods one to three, the post-treatment is ethyl acetate extraction for 3-5 times, concentration, and purification by silica gel column chromatography (n-hexane / ethyl acetate system).

[0032] Further, in the method one, the basic reagent is one or more of triethylamine, diethylamine, pyridine, DMAP, N,N-diisopropylethylamine.

[0033] Still further, in the method three, the condensing agent is one or more of dicyclohexyl carbodiimide (DCC), diisopropyl carbodiimide (DIC), 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDCI), 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ).

[0034] Further, Oxirapentyn B can be isolated and purified from the fermentation product of the sea source fungus Beauveria felina SYSU-MS7908, or can be synthesized.

[0035] The prior art isolates a series of Oxirapentyn heteroterpene compounds from the sea squirt-derived fungus Beauveria felina SYSU-MS7908, and finds that the compounds have good anti-inflammatory activity. The present application takes the natural product Oxirapentyn B as a raw material, acylates the 6-OH thereof to obtain a series of new chemical derivatives (carboxylic acid ester compound I, sulfonic acid ester compound II and carbonate compound III). The activity of the obtained heteroterpene derivatives is tested by taking the LPS-induced production of NO by RAW264.7 cells as an anti-inflammatory model, and the results show that the obtained new Oxirapentyn B heteroterpene derivatives all have strong anti-inflammatory activity, and the anti-inflammatory activity is stronger than that of the natural product Oxirapentyn B, and even higher than that of the positive control indomethacin.

[0036] Therefore, the present application also claims the use of the heteroterpene derivative of the natural product Oxirapentyn B in the preparation of an anti-inflammatory drug.

[0037] Further, the drug also comprises a pharmaceutically acceptable excipient.

[0038] Still further, the dosage form of the drug is an oral agent, an injection agent, an inhalation agent or an external agent.

[0039] The present application has the following beneficial effects:

[0040] The present application provides a heteroterpene derivative of the natural product Oxirapentyn B, which takes the metabolite heteroterpene Oxirapentyn B isolated from the marine fungus Beauveria felina SYSU-MS7908 as a raw material, and acylates the 6-OH thereof to obtain a series of new heteroterpene derivatives (containing carboxylic acid ester I, sulfonic acid ester II and carbonate III). These heteroterpene derivatives exhibit significant anti-inflammatory activity, can effectively inhibit the production of NO by inflammatory cells, and have low cytotoxicity and good safety, and are very suitable for being prepared into an anti-inflammatory drug. Moreover, the heteroterpene derivative of the present application takes the marine fungus-derived heteroterpene Oxirapentyn B as a raw material, has the characteristics of microbial scale fermentation, simple production process, short cycle and low product cost, and has a broad application prospect. DETAILED DESCRIPTION

[0041] The present application will be further described below in combination with specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and equipment used in the present application are conventional reagents, methods and equipment in the technical field.

[0042] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0043] Example 1 A method for preparing a heteroterpene derivative of natural product Oxirapentyn B

[0044] The fermentation was carried out by using sea squirt symbiotic fungus strain Beauveria felina SYSU-MS7908 (preserved in Guangdong Microbial Culture Collection Center on June 28, 2018, with the preservation number of GDMCC No: 61059, and the address of preservation is No. 59, Building 5, Guangzhou, China, which has been disclosed and protected in Chinese patent application CN112646729A, and the public can obtain it) to obtain natural product Oxirapentyn B by separation and purification of the fermentation product. The acyl reaction chemical derivation was carried out by using natural product Oxirapentyn B as raw material to obtain new heteroterpene derivatives I, II and III.

[0045] 1. The preparation process of natural product Oxirapentyn B is as follows:

[0046] The sea squirt-derived fungus Beauveria felina SYSU-MS7908 was cultured in 10 kg of sterilized solid rice medium (100 parts of rice, 100 parts of 3% sea salt water, and 3 parts of peptone) at 121℃ for 20 min at room temperature for 28 days. The obtained fermentation medium was extracted by soaking with organic solvents (methanol or acetone), and then extracted with ethyl acetate / water. The obtained ethyl acetate extract was further purified by silica gel column chromatography with petroleum ether / ethyl acetate as the mobile phase system, and the 20% volume fraction of ethyl acetate / petroleum ether was collected. ODS-C18 reverse phase chromatography was used to obtain gram-level natural product Oxirapentyn B (OXB) raw material.

[0047] The natural product Oxirapentyn B can undergo 6-OH acylation reaction with derivatization reagents (such as acyl chloride, acid anhydride and organic carboxylic acid) to form heteroterpene derivatives. Further concentration and column chromatography can obtain natural product Oxirapentyn B heteroterpene derivatives B1-B29 with carboxylic acid ester structure I, carbonate structure II or sulfonic acid ester structure III.

[0048] 2. The preparation process of natural product Oxirapentyn B derivative is as follows:

[0049] (a) Acyl chloride method: 1 eq of Oxirapentyn B (OXB) was dissolved in anhydrous dichloromethane (DCM) with 0.2 eq of 4-dimethylaminopyridine (DMAP) as catalyst and 3 eq of acyl chloride reagent (different acyl chloride reagents were selected according to different products: ClCO-R1, ClSO2-R2 or ClCO2-R3) was added. The mixture was stirred for 10 min in an ice bath and then stirred for 1-5 h at room temperature. TLC detection showed that the OXB starting material point disappeared. The reaction was quenched by adding saturated brine, and extracted with an equal volume of ethyl acetate four times. The target derivative was obtained by purification through flash silica gel column chromatography (n-hexane / ethyl acetate system).

[0050] (b) Acid anhydride method: 1 eq of Oxirapentyn B (OXB) was dissolved in anhydrous dichloromethane (DCM) with 0.2 eq of DMAP as catalyst and 1.2 eq of acid anhydride reagent (different acid anhydride reagents were selected according to different products: O(CO-R1)2, O(SO2-R2)2 or O(CO2-R3)2) was added. The mixture was stirred for 1-5 h at room temperature. TLC detection showed that the OXB starting material point disappeared. The reaction was quenched by adding saturated brine, and extracted with an equal volume of ethyl acetate four times. The target derivative was obtained by purification through flash silica gel column chromatography (n-hexane / ethyl acetate system).

[0051] (c) Carboxylic acid method: 1 eq of Oxirapentyn B (OXB) was dissolved in anhydrous dichloromethane (DCM) with 0.2 eq of DMAP as catalyst and 1.2 eq of dicyclohexyl carbodiimide (DCC) as condensing agent. 2 eq of organic carboxylic acid reagent (different organic acid reagents were selected according to different products: R1COOH, R2SO3H or R3CO3H) was added. The mixture was first stirred for 5 min in an ice bath and then stirred for 1-5 h at room temperature. TLC detection showed that the OXB starting material point disappeared. The reaction was quenched by adding saturated brine, and extracted with an equal volume of ethyl acetate four times. The target derivative was obtained by purification through flash silica gel column chromatography (n-hexane / ethyl acetate system).

[0052] According to the above method, the natural product Oxirapentyn B and 32 natural product Oxirapentyn B heteroterpene derivatives were prepared, and the specific structures are as follows:

[0053]

[0054] Example 2: Physicochemical property test data of natural product Oxirapentyn B heteroterpene derivative

[0055] The physicochemical property data of natural product Oxirapentyn B heteroterpene derivative are as follows:

[0056] Propionyloxirapentyn B (B1): Method (a), white solid (14 mg). 1 H NMR (400 MHz, CDC13) δ 5.76 (s, 1H, H-6), 5.30 (s, 1H, H-4’a), 5.27 (d, J = 1.4 Hz, 1H, H-4’b), 4.90 (t, J = 2.5 Hz, 1H, H-2), 4.45 (s, 1H, H-9), 3.39 (s, 1H, H-8), 2.84 (s, 1H, H-5), 2.53–2.46 (m, 2H, H-13), 2.45 (m, 1H, H-3a), 2.08 (s, 3H, H-1”), 1.83 (d, J = 0.8 Hz, 3H, H-5’), 1.46 (dd, J = 10.4, 3.0 Hz, 1H, H-3b), 1.42 (s, 3H, H-11), 1.28 (d, J = 12.6 Hz, 3H, H-10), 1.22 (t, J = 8.1 Hz, 4H, H-14). 13 C NMR (100 MHz, CDC13) δ 172.8 (C=0, C-12), 170.2 (C=0, C-1”), 125.6 (C, C-3’), 124.1 (CH2, C-4’), 86.0 (C, C-2’), 83.5 (C, C-1’), 74.5 (C, C-1), 73.5 (CH, C-2), 63.6 (CH, C-6), 63.5 (CH, C-9), 60.5 (CH, C-8), 58.5 (CH, C-5), 53.2 (C, C-4), 50.0 (C, C-7), 32.5 (CH2, C-3), 27.7 (CH2, C-13), 25.5 (CH3, C-10), 23.0 (CH3, C-5’), 21.7 (CH3, C-11), 20.9 (CH3, C-2”), 9.2 (CH3, C-14). HR-ESIMS: calcd. for C 21 H 27 O7[M+H] + 391.17513, found: 391.17458.

[0057] Valeryloxirapentyn B (B2): Method (a), white solid (15 mg). 1H NMR (400 MHz, CDC13) δ 5.77 (s, 1H, H-6), 5.32 (d, J = 0.7 Hz, 1H, H-4’a), 5.28 (s, 1H, H-4’b), 4.91 (t, J = 2.4 Hz, 1H, H-2), 4.46 (s, 1H, H-9), 3.40 (s, 1H, H-8), 2.84 (s, 1H, H-5), 2.50 (dd, J = 14.9, 2.4 Hz, 1H, H-3a), 2.47 - 2.37 (m, 2H, H-13), 2.09 (s, 3H, H-1”), 1.84 (s, 3H, H-5’), 1.69 (dd, J = 15.5, 7.6 Hz, 2H, H-14), 1.47 (dd, J = 10.2, 2.7 Hz, 1H, H-3b), 1.43 (s, 3H, H-11), 1.41 - 1.35 (m, 2H, H-15), 1.28 (s, 3H, H-10), 0.92 (t, J = 7.3 Hz, 3H, H-16). 13 C NMR (100 MHz, CDC13) δ 172.2 (C=0, C-12), 170.2 (C=0, C-1”), 125.6 (C, C-3’), 124.2 (CH2, C-4’), 86.0 (C, C-2’), 83.5 (C, C-1’), 74.5 (C, C-1), 73.5 (CH, C-2), 63.5 (CH, C-6), 63.5 (CH, C-9), 60.5 (CH, C-8), 58.5 (CH, C-5), 53.2 (C, C-4), 50.0 (C, C-7), 34.1 (CH2, C-13), 32.5 (CH2, C-3), 27.1 (CH2, C-14), 25.5 (CH3, C-10), 23.0 (CH3, C-5’), 22.4 (CH3, C-15), 21.7 (CH3, C-11), 21.0 (CH3, C-2”), 13.8 (CH3, C-16). HR-ESIMS: calcd. for C 23 H 30 O7Na[M+Na] + 441.18837 found: 441.18823.

[0058] Heptanoyloxirapentyn B (B3): Method (a), white oil (13 mg). 1H NMR (400 MHz, CDC13) δ 5.76 (s, 1H, H-6), 5.31 (dt, J = 5.8, 2.9 Hz, 1H, H-4’a), 5.29-5.27 (m, 1H, H-4’b), 4.93-4.88 (m, 1H, H-2), 4.45 (s, 1H, H-9), 3.40 (s, 1H, H-8), 2.84 (s, 1H, H-5), 2.53-2.46 (m, 1H, H-3a), 2.39 (tt, J = 22.8, 7.5 Hz, 2H, H-13), 2.08 (s, 3H, H-1”), 1.83 (s, 3H, H-5’), 1.74-1.65 (m, 2H, H-14), 1.46 (dd, J = 10.1, 3.1 Hz, 1H, H-3b), 1.43 (s, 3H, H-11), 1.39-1.28 (m, 6H, H-15, H-16, H-17), 1.26 (d, J = 7.9 Hz, 3H, H-10), 0.88 (t, J = 6.8 Hz, 3H, H-18). 13 C NMR (100 MHz, CDC13) δ 172.2 (C=0, C-12), 170.2 (C=0, C-1”), 125.6 (C, C-3’), 124.14 (CH2, C-4’), 86.00 (C, C-2’), 83.56 (C, C-1’), 74.51 (C, C-1), 73.49 (CH, C-2), 63.56 (CH, C-6), 63.5 (CH, C-9), 60.5 (CH, C-8), 58.5 (CH, C-5), 53.2 (C, C-4), 50.1 (C, C-7), 34.4 (CH2, C-13), 32.6 (CH2, C-3), 31.6 (CH2, C-14), 29.0 (CH2, C-15), 25.5 (CH3, C-10), 25.0 (CH2, C-16), 23.1 (CH3, C-5’), 22.6 (CH3, C-17), 21.7 (CH3, C-11), 21.0 (CH3, C-2”), 14.1 (CH3, C-18). HR-ESIMS: calcd. for C 25 H 35 O7[M+H] + 447.23773, found: 447.23736.

[0059] Myristyloxirapentyn B (B4): Method (a), white oil (19 mg). 1H NMR (400 MHz, CDC13) δ 5.76 (s, 1H, H-6), 5.31 (t, J = 3.3 Hz, 1H, H-4’b), 5.28 (dd, J = 5.6, 4.0 Hz, 1H, H-4’a), 4.91 (t, J = 3.0 Hz, 1H, H-2), 4.45 (s, 1H, H-9), 3.39 (s, 1H, H-8), 2.84 (s, 1H, H-5), 2.49 (dt, J = 11.9, 4.6 Hz, 1H, H-3a), 2.45 - 2.37 (m, 2H, H-13), 2.09 (s, 3H, H-1”), 1.83 (s, 3H, H-5’), 1.74 - 1.66 (m, 2H, H-14), 1.45 (dd, J = 10.1, 3.1 Hz, 1H, H-3b), 1.43 (s, 3H, H-11), 1.25 (m, 20H, H-15 - H-24), 1.26 (s, 3H, H-10), 0.88 (t, J = 6.8 Hz, 5H, H-25). 13 C NMR (100 MHz, CDC13) δ 172.2 (C=0, C-12), 170.2 (C=0, C-1”), 125.6 (C, C-3’), 124.2 (CH2, C-4’), 86.0 (C, C-2’), 83.6 (C, C-1’), 74.5 (C, C-1), 73.5 (CH, C-2), 63.6 (CH, C-6), 63.5 (CH, C-9), 60.5 (CH, C-8), 58.5 (CH, C-5), 53.2 (C, C-4), 50.1 (C, C-7), 34.4 (CH2, C-13), 32.6 (CH2, C-3), 32.1 (CH2, C-14), 29.9 (CH2, C-15), 29.8 (CH2, C-16), 29.8 (2 x CH2, C-17, C-18), 29.8 (CH2, C-19), 29.6 (CH2, C-19), 29.5 (CH2, C-20), 29.4 (CH2, C-21), 29.4 (CH2, C-22), 25.5 (CH3, C-10), 25.0 (CH2, C-23), 23.1 (CH3, C-5’), 22.8 (CH2, C-24), 21.7 (CH3, C-11), 21.0 (CH3, C-2”), 14.3 (CH2, C-25). HR-ESIMS: calcd. for C 32 H 49 O7Na[M+Na] + 567.32922, found: 567.32847.

[0060] Phenylbutyryloxirapentyn B (B5): Method (c), white solid (8 mg). 1 H NMR (400 MHz, CDC13) δ 7.27 (dd, J = 9.3, 4.1 Hz, 2H), 7.22 - 7.15 (m, 4H), 5.77 (d, J = 9.8 Hz, 1H), 5.27 (d, J = 0.6 Hz, 1H), 5.26 - 5.23 (m, 1H), 4.89 (t, J = 3.0 Hz, 1H), 4.45 (s, 1H), 3.39 (d, J = 8.9 Hz, 1H), 2.81 (d, J = 6.1 Hz, 1H), 2.70 (t, J = 7.5 Hz, 3H), 2.52 - 2.45 (m, 2H), 2.44 - 2.39 (m, 1H), 2.05 (dd, J = 14.6, 7.4 Hz, 3H), 1.95 (s, 3H), 1.79 (s, 3H), 1.46 - 1.43 (m, 1H), 1.42 (d, J = 3.3 Hz, 3H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.8, 21.7, 23.0, 25.5, 26.5, 32.5, 33.5, 35.2, 50.0, 53.2, 58.4, 60.4, 63.5, 63.6, 73.4, 74.5, 83.5, 86.0, 124.2, 125.5, 126.3, 128.6, 128.6, 141.1, 170.2, 171.8. HR-ESIMS: calcd. for C 28 H 32 O7 Na[M+Na] + 503.20402, found: 503.20353.

[0061] 3-Carboxypropionyloxirapentyn B (B6): Method (b), white solid (7 mg). 1 H NMR (400 MHz, CDC13) δ 5.39 (s, 1H), 5.32 (s, 1H), 4.89 (t, J = 2.9 Hz, 1H), 4.46 (s, 2H), 3.39 (s, 1H), 3.00 (s, 1H), 2.49 (dd, J = 14.7, 2.6 Hz, 1H), 2.12 (s, 3H), 1.88 (s, 3H), 1.49 (dd, J = 14.7, 3.1 Hz, 1H), 1.42 (s, 3H), 1.25 (s, 3H). 13C NMR (100 MHz, CDC13) δ 20.9, 21.7, 23.2, 25.5, 32.6, 51.8, 53.1, 60.2, 60.3, 63.9, 64.0, 73.8, 74.7, 84.0, 86.7, 124.4, 125.5, 170.5. HR-ESIMS: calcd. for C 22 H 26 O9 Na[M+Na] + 457.1469, found: 457.14678.

[0062] 3-Methyl-2-butenoyloxirapentyn B (B7): Method (a), white solid (13 mg). 1 H NMR (400 MHz, CDC13) δ 5.81 (d, J = 7.0 Hz, 2H), 5.25 (d, J = 10.0 Hz, 2H), 4.89 (t, J = 2.8 Hz, 1H), 4.46 (s, 1H), 3.40 (s, 1H), 2.88 (s, 1H), 2.49 (dd, J = 14.7, 2.6 Hz, 1H), 2.25 (s, 3H), 2.06 (s, 3H), 1.94 (s, 3H), 1.85 (d, J = 12.7 Hz, 1H), 1.80 (s, 3H), 1.48 - 1.45 (m, 1H), 1.42 (s, 3H), 1.27 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.5, 20.7, 21.6, 22.8, 25.4, 27.6, 32.4, 50.1, 53.0, 58.4, 60.3, 62.9, 63.5, 73.5, 74.3, 83.6, 86.0, 115.0, 123.6, 125.6, 159.7, 164.6, 170.2. HR-ESIMS: calcd. for C 23 H 28 O7 Na[M+Na] + 439.17272, found: 439.17282.

[0063] Benzoyloxirapentyn B (B8): Method (a), white solid (17 mg). 1H NMR (400 MHz, CDC13) δ 8.19 (d, J = 7.4 Hz, 2H), 7.61 (t, J = 7.4 Hz, 1H), 7.47 (t, J = 7.7 Hz, 2H), 5.95 (d, J = 15.5 Hz, 1H), 5.12 (s, 1H), 5.09 (s, 1H), 4.91 (t, J = 2.8 Hz, 1H), 4.51 (s, 1H), 3.47 (s, 1H), 3.03 (s, 1H), 2.52 (dd, J = 14.8, 2.5 Hz, 1H), 1.74 (s, 3H), 1.63 (s, 3H), 1.45 (d, J = 7.0 Hz, 3H), 1.42 (d, J = 3.1 Hz, 1H), 1.29 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.5, 21.7, 22.8, 25.6, 32.6, 50.1, 53.3, 58.2, 60.4, 63.6, 64.7, 73.5, 74.6, 83.4, 86.5, 123.8, 125.4, 128.7, 129.4, 130.1, 133.8, 165.1, 170.3. HR-ESIMS: calcd. for C 25 H 27 O7[M+H] + 439.17513, found: 439.17445.

[0064] P-Bromobenzoyloxirapentyn B (B9): Method (a), white solid (16 mg). 1 H NMR (400 MHz, CDC13) δ 8.19 (d, J = 7.4 Hz, 2H), 7.61 (t, J = 7.4 Hz, 1H), 7.47 (t, J = 7.7 Hz, 2H), 5.95 (d, J = 15.5 Hz, 1H), 5.12 (s, 1H), 5.09 (s, 1H), 4.91 (t, J = 2.8 Hz, 1H), 4.51 (s, 1H), 3.47 (s, 1H), 3.03 (s, 1H), 2.52 (dd, J = 14.8, 2.5 Hz, 1H), 1.74 (s, 3H), 1.63 (s, 3H), 1.45 (d, J = 7.0 Hz, 3H), 1.42 (d, J = 3.1 Hz, 1H), 1.29 (s, 3H). 13C NMR (100 MHz, CDC13) δ 20.7, 21.8, 22.8, 25.6, 32.6, 50.0, 53.4, 58.0, 60.4, 63.6, 65.0, 73.5, 74.6, 83.2, 86.6, 124.0, 125.3, 128.3, 129.1, 131.6, 132.1, 164.5, 170.1. HR-ESIMS: calcd. for C 25 H 25 O7Br Na[M+Na] + 539.06759, found: 539.06736.

[0065] P-nitrobenzoyloxirapentyn B (B10): Method (a), white solid (12 mg). 1 H NMR (400 MHz, CDC13) δ 8.39 - 8.31 (m, 4H), 5.99 (s, 1H), 5.17 - 5.12 (m, 1H), 5.09 (d, J = 0.5 Hz, 1H), 4.93 (t, J = 3.0 Hz, 1H), 4.53 (s, 1H), 3.48 (s, 1H), 3.03 (s, 1H), 2.54 (dd, J = 14.9, 2.9 Hz, 1H), 1.78 (s, 3H), 1.64 (s, 3H), 1.47 (d, J = 3.2 Hz, 1H), 1.46 (s, 3H), 1.30 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.7, 21.8, 22.8, 25.6, 32.6, 49.9, 53.5, 57.8, 60.4, 63.5, 65.7, 73.6, 74.7, 83.1, 86.7, 123.9, 124.2, 125.2, 131.2, 134.7, 151.1, 163.3, 169.9. HR-ESIMS: calcd. for C 25 H 26 NO9[M+H] + 484.16021, found: 484.15947.

[0066] P-cyanobenzoyloxirapentyn B (B11): Method (a), white solid (12 mg). 1H NMR (400 MHz, CDC13) δ 8.39 - 8.31 (m, 4H), 5.99 (s, 1H), 5.17 - 5.12 (m, 1H), 5.09 (d, J = 0.5 Hz, 1H), 4.93 (t, J = 3.0 Hz, 1H), 4.53 (s, 1H), 3.48 (s, 1H), 3.03 (s, 1H), 2.54 (dd, J = 14.9, 2.9 Hz, 1H), 1.78 (s, 3H), 1.64 (s, 3H), 1.47 (d, J = 3.2 Hz, 1H), 1.46 (s, 3H), 1.30 (s, 3H). 13 CNMR (100 MHz, CDC13) δ 20.7, 21.8, 22.8, 25.6, 32.6, 49.9, 53.5, 57.8, 60.4, 63.5, 65.7, 73.6, 74.7, 83.1, 86.7, 123.9, 124.2, 125.2, 131.2, 134.7, 151.1, 163.3, 169.9. HR-ESIMS: calcd. for C 25 H 26 NO9 [M+H] + 484.16021, found: 484.15947.

[0067] Ortho hydroxybenzoyloxirapentyn B (B12): Method (a), white solid (6 mg). 1 H NMR (400 MHz, CDC13) δ 8.39 - 8.31 (m, 4H), 5.99 (s, 1H), 5.17 - 5.12 (m, 1H), 5.09 (d, J = 0.5 Hz, 1H), 4.93 (t, J = 3.0 Hz, 1H), 4.53 (s, 1H), 3.48 (s, 1H), 3.03 (s, 1H), 2.54 (dd, J = 14.9, 2.9 Hz, 1H), 1.78 (s, 3H), 1.64 (s, 3H), 1.47 (d, J = 3.2 Hz, 1H), 1.46 (s, 3H), 1.30 (s, 3H). 13C NMR (100 MHz, CDC13) δ 20.7, 21.8, 22.7, 25.6, 32.6, 49.9, 53.5, 57.9, 60.4, 63.6, 65.0, 73.5, 74.7, 83.0, 86.9, 111.8, 118.0, 119.5, 124.1, 125.3, 130.3, 136.7, 162.1, 168.6, 170.3. HR-ESIMS: calcd. for C 25 H 27 O8[M+H] + 455.17004, found: 455.16956.

[0068] Ethylsulfonylpoxirapentyn B (B13): Method (a), white solid (12 mg). 1 H NMR (400 MHz, CDC13) δ 5.37 (s, 1H), 5.36 - 5.34 (m, 1H), 5.32 (s, 1H), 4.92 (t, J = 3.0 Hz, 1H), 4.47 (s, 1H), 3.44 (d, J = 8.1 Hz, 1H), 3.32 (q, J = 7.4 Hz, 2H), 3.20 (s, 1H), 2.50 (dd, J = 14.8, 2.8 Hz, 1H), 2.16 (s, 3H), 1.87 (s, 3H), 1.58 - 1.50 (m, 4H), 1.48 (dd, J = 14.8, 3.1 Hz, 1H), 1.42 (s, 3H), 1.26 (d, J = 4.6 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ 8.3, 20.9, 21.7, 23.0, 25.5, 32.5, 46.6, 49.4, 53.5, 58.4, 60.7, 63.3, 71.3, 73.1, 74.8, 83.6, 86.4, 124.9, 125.4, 170.9. HR-ESIMS: calcd. for C 20 H 27 O8S[M+H] + 427.14211, found: 427.14170.

[0069] Tosyloxirapentyn B (B14): Method (a), white solid (12 mg). 1H NMR (400 MHz, CDC13) δ 7.87 (d, J = 8.3 Hz, 2H), 7.33 (d, J = 8.1 Hz, 2H), 5.28 - 5.24 (m, 1H), 5.23 - 5.20 (m, 1H), 5.16 (s, 1H), 4.90 (t, J = 3.0 Hz, 1H), 4.43 (s, 1H), 3.37 (d, J = 1.4 Hz, 1H), 3.21 (s, 1H), 2.51 - 2.45 (m, 1H), 2.44 (s, 3H), 2.16 (s, 3H), 1.75 - 1.72 (m, 3H), 1.46 (dd, J = 14.7, 3.2 Hz, 1H), 1.40 (s, 3H), 1.25 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 21.0, 21.7, 21.9, 22.9, 25.5, 32.5, 49.5, 53.6, 58.1, 60.8, 63.3, 71.1, 73.2, 74.7, 83.1, 86.1, 124.3, 125.6, 128.5, 130.2, 132.8, 145.6, 170.9. HR-ESIMS: calcd for C 25 H 28 O8S Na[M+Na] + 511.13971, found: 511.13924.

[0070] Phenylsulfonyloxirapentyn B (B15): Method (a), white solid (11 mg). 1 H NMR (400 MHz, CDC13) δ 8.04 - 7.95 (m, 2H), 7.71 - 7.61 (m, 1H), 7.55 (dd, J = 10.7, 4.8 Hz, 2H), 5.26 (dd, J = 3.2, 1.6 Hz, 1H), 5.24 - 5.21 (m, 1H), 5.20 (s, 1H), 4.90 (t, J = 3.0 Hz, 1H), 4.43 (s, 1H), 3.37 (d, J = 1.4 Hz, 1H), 3.22 (s, 1H), 2.48 (dd, J = 14.7, 2.8 Hz, 1H), 2.15 (s, 3H), 1.75 - 1.71 (m, 3H), 1.45 (dd, J = 14.7, 3.2 Hz, 1H), 1.40 (s, 3H), 1.25 (s, 4H). 13C NMR (100 MHz, CDC13) δ 20.9, 21.7, 23.0, 25.4, 32.5, 49.5, 53.7, 58.0, 60.8, 63.2, 71.3, 73.2, 74.7, 83.0, 86.2, 124.6, 125.5, 128.5, 129.5, 134.4, 135.8, 170.8. HR-ESIMS: calcd for C 24 H 27 O8S[M+H] + 475.14211, found: 475.14202.

[0071] 2-Naphthylsulfonyloxirapentyn B (B16): Method (a), white solid (15 mg). 1 H NMR (400 MHz, CDC13) δ 8.57 (s, 1H), 7.99 (d, J = 7.9 Hz, 1H), 7.97 - 7.94 (m, 2H), 7.92 (d, J = 8.8 Hz, 1H), 7.73 - 7.68 (m, 1H), 7.68 - 7.63 (m, 1H), 5.27 (s, 1H), 5.09 - 5.03 (m, 1H), 4.99 (dd, J = 1.7, 0.9 Hz, 1H), 4.91 (t, J = 3.0 Hz, 1H), 4.43 (s, 1H), 3.36 (d, J = 1.5 Hz, 1H), 3.27 (s, 1H), 2.48 (dd, J = 14.7, 2.8 Hz, 1H), 2.17 (s, 3H), 1.53 - 1.51 (m, 3H), 1.47 (dd, J = 14.7, 3.2 Hz, 1H), 1.40 (s, 3H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 21.0, 21.7, 22.7, 25.5, 32.5, 49.5, 53.7, 58.2, 60.8, 63.3, 71.6, 73.2, 74.7, 82.8, 86.2, 123.0, 124.4, 125.3, 128.1, 128.1, 129.7, 129.9, 129.9, 130.4, 132.1, 132.6, 135.7, 170.9. HR-ESIMS: calcd for C 28 H 29 O8S[M+H] + 525.15776, found: 525.15775.

[0072] 1 -Naphthylsulfonyloxirapentyn B (B17): Method (a), white solid (10 mg). 1 H NMR (400 MHz, CDC13) δ 8.75 (d, J = 8.6 Hz, 1H), 8.36 (d, J = 7.4 Hz, 1H), 8.13 (d, J = 8.2 Hz, 1H), 7.92 (d, J = 8.2 Hz, 1H), 7.69 (t, J = 7.8 Hz, 1H), 7.59 (dd, J = 15.3, 7.7 Hz, 2H), 5.34 (s, 1H), 4.94 (d, J = 1.4 Hz, 1H), 4.91 (d, J = 2.7 Hz, 1H), 4.76 (s, 1H), 4.43 (s, 1H), 3.37 (s, 1H), 3.24 (s, 1H), 2.47 (dd, J = 14.7, 2.7 Hz, 1H), 2.07 (s, 3H), 1.47 - 1.42 (m, 1H), 1.40 (s, 6H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.9, 21.7, 22.5, 25.5, 32.5, 49.6, 53.6, 58.4, 60.9, 63.3, 71.9, 73.0, 74.7, 82.6, 86.2, 124.2, 124.4, 124.6, 125.5, 127.4, 128.8, 129.0, 129.0, 130.6, 131.9, 134.5, 135.9, 170.9. HR-ESIMS: calcd. for C 28 H 28 O8S Na[M+Na] + 547.13971, found: 547.13981.

[0073] 5-(Dimethylamino)naphthalen-1-sulfonyl oxirapentyn B (B18): Method (a), white solid (13 mg). 1H NMR (400 MHz, CDC13) δ 8.60 (d, J = 8.5 Hz, 1H), 8.39 (d, J = 8.7 Hz, 1H), 8.35 (dd, J = 7.3, 0.9 Hz, 1H), 7.61 - 7.51 (m, 2H), 7.17 (d, J = 7.5 Hz, 1H), 5.33 (s, 1H), 4.98 - 4.93 (m, 1H), 4.90 (t, J = 3.0 Hz, 1H), 4.80 (d, J = 0.7 Hz, 1H), 4.42 (s, 1H), 3.37 (s, 1H), 3.23 (s, 1H), 2.87 (s, 6H), 2.46 (dd, J = 14.7, 2.8 Hz, 1H), 2.07 (s, 3H), 1.61 (d, J = 7.8 Hz, 1H), 1.44 (s, 3H), 1.40 (s, 3H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.9, 21.7, 22.6, 25.5, 32.5, 45.5, 49.6, 53.6, 58.5, 61.0, 63.3, 71.7, 73.0, 74.7, 82.7, 86.2, 115.8, 120.0, 123.2, 124.2, 124.8, 129.0, 130.2, 130.3, 130.6, 132.0, 132.2, 151.9, 170.9. HR-ESIMS: calcd. for C 30 H 34 NO8S [M+H] + 568.19996, found: 568.19981.

[0074] 8-Quinolinylsulfonyloxirapentyn B (B19): Method (a), white solid (8 mg). 1H NMR (400 MHz, CDC13) δ 9.18 (dd, J = 4.2, 1.7 Hz, 1H), 8.60 (dd, J = 7.4, 1.4 Hz, 1H), 8.26 (dd, J = 8.4, 1.7 Hz, 1H), 8.11 (dd, J = 8.2, 1.3 Hz, 1H), 7.70 - 7.63 (m, 1H), 7.58 (dd, J = 8.3, 4.3 Hz, 1H), 6.65 (s, 1H), 4.93 - 4.88 (m, 1H), 4.86 (t, J = 3.0 Hz, 1H), 4.69 - 4.63 (m, 1H), 4.44 (s, 1H), 3.36 (s, 1H), 3.28 (s, 1H), 2.46 (dd, J = 14.7, 2.8 Hz, 1H), 2.01 (s, 3H), 1.43 (dd, J = 10.6, 4.2 Hz, 1H), 1.39 (d, J = 4.7 Hz, 3H), 1.39 - 1.36 (m, 3H), 1.24 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.8, 21.7, 22.5, 25.5, 32.5, 49.9, 53.6, 59.0, 61.1, 63.4, 73.2, 73.2, 74.5, 83.3, 85.9, 122.8, 123.5, 125.1, 125.5, 129.3, 133.4, 134.9, 135.9, 136.7, 143.9, 152.1, 171.0. HR-ESIMS: calcd. for C 27 H 28 NO8S [M+H] + 526.15301, found: 526.15303.

[0075] 5,6,7,8-Tetrahydro-2-naphthylsulfonyl oxirapentyn B (B20): Method (a), white solid (16 mg). 11H NMR (400 MHz, CDCl3) δ 7.68–7.64 (m, 2H), 7.19 (d, J = 8.6 Hz, 1H), 5.25–5.22 (m, 1H), 5.22–5.19 (m, 1H), 5.17 (s, 1H), 4.89 (t, J = 3.0 Hz, 1H), 4.42 (s, 1H), 3.36 (d, J = 1.2 Hz, 1H), 3.22 (s, 1H), 2.81 (s, 4H), 2.47 (dd, J = 14.7, 2.7 Hz, 1H), 2.16 (s, 3H), 1.81 (dt, J = 6.4, 3.3 Hz, 5H), 1.75–1.72 (m, 3H), 1.46 (dd, J = 14.6, 3.1 Hz, 1H), 1.40 (s, 4H), 1.25 (s, 4H). 13 13C NMR (100 MHz, CDCl3) δ 21.0, 21.6, 22.6, 22.7, 23.0, 25.4, 29.4, 29.8, 32.5, 49.5, 53.6, 58.2, 60.8, 63.2, 71.0, 73.2, 74.6, 83.0, 86.1, 124.2, 125.3, 125.5, 129.1, 130.3, 132.5, 138.8, 144.8, 170.9. HR-ESIMS: calcd. for C 28 H 32 O8S Na [M + Na] + 551.17101, found: 551.17098.

[0076] P-Cyclohexylphenylsulfonyloxirapentyn B (B21): Method (a), white solid (12 mg). 1 1H NMR (400 MHz, CDCl3) δ 7.92–7.86 (m, 2H), 7.36 (d, J = 8.4 Hz, 2H), 5.26 (dd, J = 3.2, 1.6 Hz, 1H), 5.24–5.21 (m, 1H), 5.18 (s, 1H), 4.89 (t, J = 3.0 Hz, 1H), 4.43 (s, 1H), 3.37 (d, J = 1.3 Hz, 1H), 3.24 (s, 1H), 2.59 (d, J = 8.6 Hz, 1H), 2.47 (dd, J = 14.7, 2.7 Hz, 1H), 2.14 (s, 3H), 1.86 (d, J = 7.6 Hz, 4H), 1.75–1.72 (m, 3H), 1.47 (d, J = 3.2 Hz, 1H), 1.39 (s, 5H), 1.24 (s, 4H), 0.87 (t, J = 6.9 Hz, 1H). 13C NMR (100 MHz, CDC13) δ 21.0, 21.6, 23.0, 25.4, 26.0, 26.7, 32.5, 34.1, 44.8, 49.5, 53.6, 58.1, 60.8, 63.2, 71.0, 73.1, 74.6, 83.1, 86.2, 124.4, 125.5, 128.0, 128.7, 132.8, 155.3, 170.9. HR-ESIMS: calcd. for C 30 H 36 O8S Na[M+Na] + 579.20231, found: 579.20245.

[0077] P-benzenesulfonyloxirapentyn B (B22): Method (a), white solid (13 mg). 1 H NMR (400 MHz, CDC13) δ 8.05 (d, J = 8.5 Hz, 2H), 7.74 (d, J = 8.5 Hz, 2H), 7.59 (d, J = 7.0 Hz, 2H), 7.50 (t, J = 7.3 Hz, 2H), 7.45 (d, J = 7.1 Hz, 1H), 5.24 (s, 1H), 5.20 (d, J = 1.5 Hz, 1H), 5.19 (s, 1H), 4.92 (t, J = 2.9 Hz, 1H), 4.45 (s, 1H), 3.38 (s, 1H), 3.28 (s, 1H), 2.49 (dd, J = 14.7, 2.7 Hz, 1H), 2.18 (s, 3H), 1.69 (s, 3H), 1.48 (dd, J = 14.7, 3.1 Hz, 1H), 1.41 (s, 3H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 21.0, 21.6, 22.9, 25.4, 32.5, 49.4, 53.7, 58.1, 60.8, 63.2, 71.4, 73.2, 74.7, 83.0, 86.2, 124.5, 125.5, 127.4, 128.0, 129.0, 129.3, 134.2, 138.9, 147.4, 170.9. HR-ESIMS: calcd. for C 30 H 30 O8S Na[M+Na] + 573.15536, found: 573.15544.

[0078] P-methoxybenzenesulfonyloxirapentyn B (B23): Method (a), white solid (13 mg).1 HNMR (400 MHz, CDC13) δ 7.94 - 7.89 (m, 2H), 7.01 - 6.95 (m, 2H), 5.28 - 5.25 (m, 1H), 5.23 (dd, J = 1.8, 1.0 Hz, 1H), 5.11 (s, 1H), 4.89 (t, J = 3.0 Hz, 1H), 4.42 (s, 1H), 3.87 (s, 3H), 3.36 (d, J = 1.4 Hz, 1H), 3.22 (s, 1H), 2.47 (dd, J = 14.7, 2.8 Hz, 1H), 2.17 (s, 3H), 1.76 - 1.72 (m, 3H), 1.45 (dt, J = 7.0, 3.5 Hz, 1H), 1.39 (s, 3H), 1.25 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 21.0, 21.6, 23.0, 25.4, 32.5, HRESIMS: calcd. for C 25 H 28 O9S Na[M+Na] + 527.13462, found: 527.13474.

[0079] P-trifluoromethylbenzenesulfonyloxirapentyn B (B24): Method (a), white solid (9 mg). 1 H NMR (400 MHz, CDC13) δ 8.13 (d, J = 8.2 Hz, 2H), 7.81 (d, J = 8.3 Hz, 2H), 5.23 (dd, J = 3.2, 1.4 Hz, 2H), 5.11 (d, J = 0.6 Hz, 1H), 4.92 (t, J = 3.0 Hz, 1H), 4.45 (s, 1H), 3.37 (d, J = 1.4 Hz, 1H), 3.28 (d, J = 0.5 Hz, 1H), 2.50 (dd, J = 14.7, 2.8 Hz, 1H), 2.18 (d, J = 3.7 Hz, 3H), 1.67 - 1.64 (m, 3H), 1.48 (dd, J = 14.7, 3.1 Hz, 1H), 1.41 (s, 3H), 1.26 (s, 3H). 13C NMR (100 MHz, CDC13) δ 20.9, 21.6, 22.8, 25.4, 32.5, 49.2, 53.7, 58.0, 60.7, 63.2, 72.3, 73.1, 74.8, 82.8, 86.1, 124.9, 125.2, 126.6, 126.6, 126.7, 126.7, 129.0, 135.4, 135.8, 136.1, 136.4, 139.4, 170.8. HR-ESIMS: calcd. for C 25 H 25 F3O8SNa[M+Na] + 565.11144, found: 565.11147.

[0080] P-acetamidobenzenesulfonyloxirapentyn B (B25): Method (a), white solid (10 mg). 1 H NMR (400 MHz, CDC13) δ 7.96 - 7.91 (m, 2H), 7.71 (d, J = 8.8 Hz, 2H), 5.29 - 5.26 (m, 1H), 5.25 (s, 1H), 5.15 (s, 1H), 4.90 (t, J = 3.0 Hz, 1H), 4.43 (s, 1H), 3.37 (s, 1H), 3.22 (s, 1H), 2.48 (dd, J = 14.7, 2.7 Hz, 1H), 2.24 (s, 3H), 2.16 (s, 3H), 1.76 (s, 3H), 1.46 (dd, J = 14.7, 3.1 Hz, 1H), 1.40 (s, 3H), 1.25 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 21.0, 21.7, 23.0, 25.0, 25.5, 32.5, 49.5, 53.7, 58.0, 60.8, 63.2, 71.1, 73.2, 74.7, 83.0, 86.2, 119.3, 124.6, 125.5, 128.4, 130.0, 130.0, 143.5, 168.6, 170.9. HR-ESIMS: calcd. for C 26 H 28 O9NS[M-H] - 530.14903, found: 530.14954.

[0081] Oxirapentyn B n-propyl carbonate (B26): Method (a), white solid (12 mg). 1H NMR (400 MHz, CDC13) δ 5.58 (s, 1H), 5.33 (d, J = 0.7 Hz, 1H), 5.29 - 5.25 (m, 1H), 4.87 (t, J = 3.0 Hz, 1H), 4.45 (s, 1H), 4.22 - 4.15 (m, 2H), 3.40 (s, 1H), 2.92 (s, 1H), 2.48 (dd, J = 14.7, 2.7 Hz, 1H), 2.10 (d, J = 4.7 Hz, 3H), 1.84 (d, J = 1.1 Hz, 3H), 1.72 (dp, J = 14.1, 7.0 Hz, 2H), 1.48 (dd, J = 14.7, 3.2 Hz, 1H), 1.41 (s, 3H), 1.26 (s, 3H), 0.96 (t, J = 7.4 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ 10.2, 20.8, 21.6, 22.1, 23.0, 25.5, 32.4, 49.8, 53.3, 58.3, 60.5, 63.4, 67.4, 70.6, 73.5, 74.5, 83.0, 86.4, 124.1, 125.6, 154.6, 170.6. HR-ESIMS: calcd. for C 22 H 29 O8[M+H] + 421.18569, found: 421.18523.

[0082] Oxirapentyn B n-pentyl carbonate (B27): Method (a), white solid (12 mg). 1 H NMR (400 MHz, CDC13) δ 5.62 - 5.51 (m, 1H), 5.37 - 5.25 (m, 1H), 4.90 - 4.82 (m, 1H), 4.44 (s, 1H), 4.21 (dd, J = 12.0, 5.3 Hz, 2H), 3.47 - 3.34 (m, 1H), 2.91 (t, J = 7.5 Hz, 1H), 2.48 (dd, J = 14.7, 2.6 Hz, 1H), 2.10 (s, 3H), 1.81 (d, J = 21.8 Hz, 2H), 1.75 - 1.59 (m, 3H), 1.54 - 1.44 (m, 2H), 1.39 (d, J = 14.0 Hz, 3H), 1.37 - 1.31 (m, 4H), 1.26 (s, 3H), 0.90 (d, J = 2.6 Hz, 3H). 13CNMR (100 MHz, CDC13) δ 9.2, 20.9, 21.7, 23.0, 25.5, 27.7, 32.5, 50.0, 53.2, 58.5, 60.5, 63.5, 63.6, 73.5, 74.5, 83.5, 86.0, 124.1, 125.6, 170.2, 172.8. HR-ESIMS: calcd. for C 24 H 33 O8[M+H] + 449.21699, found: 449.21681.

[0083] Benzyl oxirapentyn B carbonate (B28): Method (a), white solid (14 mg). 1 H NMR (400 MHz, CDC13) δ 7.41 - 7.32 (m, 5H), 5.60 (s, 1H), 5.27 (d, J = 0.9 Hz, 1H), 5.25 (s, 2H), 5.23 (s, 1H), 4.85 (t, J = 3.0 Hz, 1H), 4.44 (s, 1H), 3.40 (s, 1H), 2.91 (s, 1H), 2.48 (dd, J = 14.7, 2.6 Hz, 1H), 1.98 (s, 3H), 1.79 (s, 3H), 1.46 (dd, J = 14.7, 3.2 Hz, 1H), 1.40 (s, 3H), 1.24 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.7, 21.6, 23.0, 25.5, 32.4, 49.8, 53.3, 58.2, 60.5, 63.4, 67.8, 70.6, 73.5, 74.5, 82.9, 86.5, 124.2, 125.5, 128.6, 128.9, 129.0, 134.9, 154.5, 170.6. HR-ESIMS: calcd. for C 26 H 28 O8Na[M+Na] + 491.16764, found: 491.16694.

[0084] Oxirapentyn B phenyl carbonate (B29): Method (a), white solid (14 mg). 1H NMR(400MHz,CDCl3)δ7.45–7.35(m,2H),7.33–7.27(m,1H),7.18(dt,J=9.1,2.4Hz,2H),5.68(s,1H),5.37(d,J=0.7Hz,1H),5.34–5.29(m,1H),4.91(t,J=3.0Hz,1H),4.49(s,1H),3.46(s,1H),3.02(s,1H),2.52(dd,J=14.8,2.7Hz,1H),2.15(s,3H),1.88–1.84(m,3H),1.52(dd,J=14.8,3.3Hz,1H),1.44(s,3H),1.29(s,3H). 13 C NMR(100MHz,CDCl3)δ20.9,21.6,23.0,25.5,32.4,49.7,53.4,58.1,60.5,63.4,68.2,73.5,74.6,82.9,86.7,120.8,124.5,125.6,126.5,129.8,151.2,153.0,170.6.HR-ESIMS:calcd.for C 25 H 27 O8[M+H] + 455.17004,found:455.16938.

[0085] 5-Norbornene-Endo-2-carboxyl-3-acyl oxirapentyn B(B30):Method(b),white solid(8mg). 1 H NMR(400MHz,CDCl3)δ6.31(s,2H),5.67(s,1H),5.35(s,1H),5.31(s,1H),4.88(s,1H),4.43(s,1H),3.41(d,J=9.9Hz,2H),3.32–3.25(m,1H),3.23–3.18(m,1H),2.89(s,1H),2.45(d,J=14.8Hz,1H),2.11(s,3H),2.04(s,1H),1.87(s,3H),1.55–1.47(m,2H),1.42(s,3H),1.36(d,J=8.8Hz,2H),1.27(s,3H). 13C NMR (100 MHz, CDC13) δ 20.9, 21.8, 23.1, 25.5, 32.3, 46.4, 46.9, 47.8, 48.7, 48.9, 50.1, 53.1, 58.1, 60.6, 63.5, 63.6, 73.5, 74.5, 83.8, 85.7, 124.2, 125.7, 135.1, 135.3, 170.7, 171.0, 175.8. HR-ESIMS: calcd for C 27 H 31 O9[M+H] + 499.19626, found: 499.19595.

[0086] 2-(1,3-dioxoisoindolin-2-yl)ethanesulfonylOxirapentyn B(B31): method (a), white solid (18 mg). 1 H NMR (400 MHz, CDC13) δ 7.87 (dd, J = 5.4, 3.0 Hz, 2H), 7.78 - 7.72 (m, 2H), 5.37 (s, 2H), 5.25 (d, J = 1.5 Hz, 1H), 4.91 (d, J = 2.7 Hz, 1H), 4.46 (s, 1H), 4.28 (dd, J = 12.8, 6.5 Hz, 2H), 3.79 - 3.64 (m, 2H), 3.44 (s, 1H), 3.17 (s, 1H), 2.49 (d, J = 14.7 Hz, 1H), 2.11 (s, 3H), 1.84 (d, J = 0.8 Hz, 3H), 1.46 (dd, J = 14.7, 2.7 Hz, 1H), 1.41 (s, 3H), 1.26 (s, 3H). 13 C NMR (100 MHz, CDC13) δ 20.9, 21.7, 22.9, 25.4, 32.2, 32.4, 49.2, 49.4, 53.6, 58.2, 60.6, 63.3, 72.1, 73.0, 74.8, 83.5, 86.8, 123.8, 125.0, 125.4, 131.9, 134.5, 167.5, 170.9. HR-ESIMS: calcd for C 28 H 29 NO 10 S Na[M+Na] + 594.14044, found: 594.14058

[0087] oxirapentyn B acetace (OXB acetace): 1H NMR (400 MHz, CDC13) δ H 5.71 (1H, s, H-6), 5.27 (1H, s, H-4a'), 5.24 (1H, d, J = 1.0 Hz, H-4b'), 4.85 (1H, t, J = 2.6 Hz, H-2), 4.41 (s, 1H, H-9), 3.34 (s, 1H, H-8), 2.80 (s, 1H, H-5), 2.45 (dd, J = 14.7, 2.8 Hz, 1H, H-3a), 2.14 (3H, s), 2.05 (3H, s), 1.80 (3H, s, H-5'), 1.43 - 1.40 (1H, m, H-3b), 1.38 (3H, s, H-11), 1.22 (3H, s, H-10). 13 C NMR (100 MHz, CDC13) δ C 21.7, 23.1, 25.5, 35.1, 51.7, 52.9, 60.5, 60.5, 63.6, 63.7, 72.3, 76.0, 84.2, 86.3, 124.2, 125.5. HR-ESIMS: calcd. for C 18 H 25 O5[M+H] + 321.19629, found: 321.19583.

[0088] Example 3 Anti-inflammatory activity test of natural product Oxirapenty B heteroterpene derivatives

[0089] The anti-inflammatory activity of 33 Oxirapenty heteroterpene derivatives (B1~B31, OXB and OXB acetace) prepared in Example 1 was tested by testing the amount of NO released in LPS-induced mouse macrophage RAW264.7, and the specific process was as follows:

[0090] 1. Experimental materials

[0091] Lipopolysaccharide (LPS), indomethacin (positive control), mouse macrophage (RAW264.7), DMSO, tetrazolium (MTT, 5 mg / mL), Griess method NO kit (Shanghai Biyun Tian Biotechnology Co., Ltd.).

[0092] 2. Experimental method

[0093] The compound was dissolved in DMSO to prepare a 10 mM stock solution, and when used, it was diluted with DMEM medium to the required use concentration (DMSO content was less than 0.5%).

[0094] The experimental groups were: blank group (100 μL DMEM medium), model group (1 μL LPS + 99 μL DMEM medium), positive control group (1 μL LPS + 25 μL indomethacin + 74 μL DMEM cell culture medium), sample group (1 μL LPS + 99 μL culture medium containing different concentrations of samples); wherein the concentrations of LPS and indomethacin were 100 μg / mL and 200 μg / mL, respectively.

[0095] RAW264.7 cells (1×10 5 μL) were cultured in 96-well plates at 100 μL per well, incubated at 37°C in a 5% CO2 incubator for 12 h; according to the grouping, different concentrations of samples containing lipopolysaccharide LPS (final concentration 1 μg / mL) were added to each well, and then incubated for 24 h, 50 μL of supernatant was carefully aspirated into another 96-well plate, and NO I and NO II reagents in the Griess method NO kit were added, mixed uniformly, and then incubated at room temperature for 10 min, and the absorbance value at 540 nm was measured by enzyme-labeled instrument Multiskan GO (Thermo Scientific), and the NO release level of each group of cells was calculated according to the standard curve.

[0096] The remaining 50 μL of culture solution was carefully aspirated, 100 μL of MTT solution diluted with DMEM was added, and it was placed in the incubator for 4 h; the supernatant was aspirated, 110 μL of DMSO solution was added, and shaken for 10 min, and the absorbance value at 490 nm was measured by enzyme-labeled instrument, and the survival rate of cells was evaluated.

[0097] Calculation method:

[0098] NO release inhibition rate % = (OD 模型组 - OD 样品组 ) / (OD 模型组 - OD 空白组 )×100%.

[0099] Cell survival rate % = [(average OD value measured in sample group) / (average OD value measured in control group)]×100%.

[0100] 3、Experimental results

[0101] The experimental results are shown in Table 1.

[0102] Table 1 Anti-inflammatory activity of natural product Oxirapenty B and its derivatives

[0103]

[0104]

[0105] Note: aThe anti-inflammatory results are mean ± standard deviation (n = 3), and indomethacin (Indo) is a positive control, and the cytotoxic activity is the activity of the sample on RAW264.7 cells.

[0106] As can be seen from the table, the natural product Oxirapenty B derivatives all exhibit good anti-inflammatory activity (IC 50 = 1.2-27.7 μM), stronger than the positive control indomethacin (IC 50 = 35.8 μM); compared with the natural product Oxirapenty B (OXB) with weaker anti-inflammatory activity, the anti-inflammatory activity of the derivatives is significantly enhanced, and the activity is increased by nearly 40 times, for example, the activity IC 50 of 19 compounds is lower than 10 μM, and the IC 50 values of 7 compounds (B3, B5, B7, B11, B16, B19 and B23) are less than 3 μM. Only compounds B30 and B31 have relatively poor anti-inflammatory effect. In the MTT test, except for compounds B7, B11, B14, B15, B24, B25 and B27, the other compounds have no cytotoxicity (IC 50 > 50 μM) on RAW264.7 cells.

[0107] Therefore, the natural product Oxirapenty B heteroterpene derivatives have good potential as anti-inflammatory drugs.

[0108] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are all included in the protection scope of the present application.

Claims

1. A heteroterpene derivative of the natural product Oxirapentyn B, characterized by, The heteroterpene derivative of the natural product Oxirapentyn B has any one of the following structures: said R1 is selected from C 2~15 alkyl or substituted C 1~5 alkyl, methyl substituted ethenyl, phenyl, substituted phenyl, one of the following: said R2is selected from C 1~5 alkyl, phenyl or substituted phenyl, naphthyl or substituted naphthyl, quinolinyl, tetrahydronaphthyl, cyclohexylphenyl, biphenyl, one of the group consisting of said R3 is selected from C 1~5 one of alkyl, benzyl, phenyl; The "substituted" substituent is each independently selected from phenyl, carboxyl, methyl, halogen, nitro, cyano, hydroxyl, methyl-substituted amine, methoxy, trifluoromethyl, acetamido.

2. The heteroterpene derivative of natural product Oxirapentyn B according to claim 1, characterized by, said R1is selected from any one of the following structures: -CH2CH3, -(CH2)3CH3, -(CH2)5CH3, -(CH2) 12 CH3, -(CH2)3Ph, -(CH2)2COOH, -Ph, , , , , ; R2is selected from any one of the following structures: -CH3, -CH2CH3, -PhMe, -Ph, , , , , , , , , , ; The R3 is selected from any one of the following structures: -CH3, -(CH2)2CH3, -(CH2)4CH3, -CH2Ph, -Ph.

3. Process for the preparation of the heteroterpene derivative of the natural product Oxirapentyn B according to claim 1 or 2, characterized in that, Method one specifically includes the following steps: In the presence of an organic solvent, a catalyst and a basic reagent, Oxirapentyn B is reacted with an acyl chloride reagent at room temperature until complete, and then treated to obtain; the acyl chloride reagent is ClCO-R1, ClSO2-R2 or ClCO2-R3, wherein R1, R2, R3 are defined as in claim 1 or 2.

4. Process for the preparation of the heteroterpene derivative of the natural product Oxirapentyn B according to claim 1 or 2, characterized in that, Method two specifically includes the following steps: In the presence of an organic solvent, a catalyst, Oxirapentyn B is reacted with an acid anhydride reagent at room temperature until complete, and then treated to obtain; the acid anhydride reagent is O(CO-R1)2, O(SO2-R2)2 or O(CO2-R3)2, wherein R1, R2, R3 are defined as in claim 1 or 2.

5. Process for the preparation of the heteroterpene derivative of the natural product Oxirapentyn B according to claim 1 or 2, characterized in that, Method three specifically includes the following steps: In the presence of an organic solvent, a catalyst and a condensing agent, Oxirapentyn B is reacted with an organic acid reagent at room temperature until complete, and then treated to obtain, the organic acid reagent is R1COOH, R2SO3H or R3CO3H, wherein R1, R2, R3 are defined as in claim 1 or 2.

6. The process according to any one of claims 3 to 5, characterized in that, The catalyst is selected from one or more of 4-dimethylaminopyridine, pyridine, triethylamine, diethylamine, ethylenediamine, N,N-diisopropylethylamine.

7. The method of any one of claims 3 to 5, wherein the compound is prepared by the method of any one of claims 1 to 2. The organic solvent is selected from one or more of dichloromethane, tetrahydrofuran, N,N-dimethylformamide, 1,4-dioxane, acetonitrile, toluene, chloroform, triethylamine, acetone, pyridine.

8. Use of the heteroterpene derivative of the natural product Oxirapentyn B of claim 1 or 2 in the preparation of an anti-inflammatory drug.

9. Use of the heteroterpene derivative of the natural product Oxirapentyn B for the preparation of anti-inflammatory medicaments, characterized in that, The heteroterpene derivative of the natural product Oxirapentyn B has the following structure: 。

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