Eucalyptane-guaiane sesquiterpene dimer as well as preparation method and application thereof

Abstract

The invention provides a novel eudecane-guaiane sesquiterpene dimer as shown in a structural formula as well as a pharmaceutical composition, a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method provided by the invention comprises the following steps: by taking 6 different structures of cinemanane lactone as raw materials, carrying out Diels-Alder reaction / deprotection on the raw materials and guaiane dienes to prepare 6 cinemanane-guaiane sesquiterpene dimers, and further carrying out structural modification to obtain 6 derivatives; or preparing a germacane-guaiane dimer by taking costunolide as a raw material, and further performing intramolecular cyclization to obtain 15 eudecane-guaiane sesquiterpene dimers. The eudecane-guaiane sesquiterpene dimer provided by the invention has inhibitory activity on human hepatoma cell lines HepG2, Huh7 and SK-Hep-1, can form a pharmaceutical composition with a pharmaceutically acceptable carrier, and can be used for preparing anti-hepatoma drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, this invention relates to a novel eucalyptane-guaiacane sesquiterpene dimer, its preparation method, a pharmaceutical composition for treating liver cancer with the compound as an active ingredient, and the application of the compound and its pharmaceutical composition in the preparation of anti-liver cancer drugs. Background Technology

[0002] Liver cancer is one of the most common malignant tumors, including primary and secondary liver cancer, with hepatocellular carcinoma (HCC) accounting for over 90%. Globally, HCC ranks sixth in incidence among malignant tumors, with over 800,000 new cases diagnosed annually. Liver cancer is characterized by its insidious onset, long latency period, high invasiveness, and easy metastasis. Its malignancy is also high; for patients with unresectable or advanced disease, the 5-year survival rate is only 13%. Currently, clinically available drugs for treating liver cancer mainly include four small-molecule synthetic drugs that act as tyrosine kinase inhibitors: sorafenib, regoratinib, and lenvatinib; and three antibody drugs: nivolumab, pembrolizumab, and ramucirumab. These drugs have played a significant role in the clinical treatment of liver cancer, but they still have drawbacks such as a low objective response rate (<20%), susceptibility to drug resistance, intolerance in some liver cancer patients, significant toxic side effects, and high cost, resulting in unsatisfactory efficacy.

[0003] Eucalyptane-guaiacol sesquiterpene dimers are a class of structurally complex natural products with a backbone of at least 30 carbon atoms, formed by the linkage of a eucalyptane sesquiterpene unit and a guaiacol sesquiterpene unit. Many proteins responsible for cellular function exist in dimer form or require dimerization for activation before mediating certain signaling pathways. Targeting both monomers of a dimer protein is an important strategy in drug research. Because sesquiterpene dimers have the potential to act on both monomers of a dimer protein simultaneously, the design and synthesis of eucalyptane-guaiacol sesquiterpenes are of great significance.

[0004] There are no reports on the derivatization and synthesis of eucalyptane-guaiacane sesquiterpene dimers in the prior art, especially those containing heteroatoms such as fluorine, chlorine, bromine, iodine, sulfur, and selenium. There are also no reports on pharmaceutical compositions containing them as active ingredients, nor on the application of eucalyptane-guaiacane sesquiterpene dimers and their pharmaceutical compositions in the preparation or treatment of liver cancer drugs. Summary of the Invention

[0005] The purpose of this invention is to provide novel eucalyptane-guaiacane sesquiterpene dimers 1-26, their pharmaceutical compositions, preparation methods, and applications. The preparation method of this invention uses guaiacane dienes and eucalyptol or eucalyptol as raw materials to prepare novel eucalyptane-guaiacane sesquiterpene dimers. The method uses readily available raw materials, is easy to operate, and is suitable for industrial production. This method has good substrate universality and can be used to prepare eucalyptane-guaiacane sesquiterpene dimers with various functional group substitutions.

[0006] To achieve the above-mentioned objectives of the present invention, the present invention provides the following technical solution:

[0007] This invention provides a novel compound 1-26, as shown in the following structural formula, namely, guaiacol-eucalyptane sesquiterpene dimer 1-26.

[0008]

[0009] The present invention also provides a method for preparing guaiac-guaiac sesquiterpene dimers 1-26 (compounds 1-26) as shown in the structural formula. The method includes preparing guaiac-guaiac sesquiterpene dimers 1-6 (compounds 1-6) by reacting guaiac diene C1 with guaiac-type sesquiterpenes α-cyclocostunolide, β-cyclocostunolide, γ-cyclocostunolide, santamarin, reynosin, and magnolialide via Diels-Alder reaction / demethylamine removal; or preparing guaiac-guaiac dimers from guaiac hydroxyl lactone as a raw material, and further subjecting the guaiac-guaiac dimers to intramolecular cyclization to obtain guaiac-guaiac sesquiterpene dimers 7-21 (compounds 7-21), and then preparing guaiac-guaiac sesquiterpene dimer derivatives 22-26 (compounds 22-26) by esterification, oxidation, epoxidation, etc.

[0010]

[0011] The synthesis method of eucalyptane-guaiacane sesquiterpene dimers mainly includes the following 8 key reaction steps:

[0012] Preparation of compounds 1-6: Eucalyptane-guaiacane sesquiterpene dimers 1-6 (compounds 1-6) were prepared by reacting guaiacane-type dienes with eucalyptane-type sesquiterpenes α-cyclocostunolide, β-cyclocostunolide, γ-cyclocostunolide, santamarin, reynosin, and magnolialide under appropriate conditions via Diels-Alder reaction / dedimethylamine removal. The appropriate conditions for the Diels-Alder reaction were that the dienes reacted with diephiles under solvent-free conditions with heating.

[0013] Preparation of compounds 7-8: Using tetrahydrofuran and nitromethane as solvents, compound C2 reacted with N-fluorobisbenzenesulfonamide (NFSI) at 0°C. o Compounds 7 and 8 were prepared by reacting at room temperature (C);

[0014] Preparation of compounds 9-17: Using tetrahydrofuran and water as solvents, compound C2 reacts with N-chlorosuccinimide (NCS), N-bromosuccinimide (NCS), or N-iodosuccinimide (NIS) at 0°C. o Compounds 9-17 were prepared by reacting at room temperature (C).

[0015] Preparation of compounds 18 and 19: Compound C2 was reacted with dimethyl(methylthio)sulfonium tetrafluoroborate (DMTSF) at 0°C. o Compounds 18 and 19 were prepared by reacting at room temperature (C);

[0016] Preparation of compounds 20 and 21: Compound C2 was reacted with phenylselenotrifluoromethane sulfonate (PhSeOTf) in dichloromethane as a solvent at 0°C. o Compounds 20 and 21 were prepared by reaction under C.

[0017] Preparation of compounds 22 and 23: Compound 4 was reacted with acetic anhydride or benzoic anhydride in the presence of 4-dimethylaminopyridine (DMAP) to prepare compounds 22 and 23;

[0018] Preparation of compound 24: Compound 4 was prepared by oxidation reaction in the presence of Dess-Martin periodinane DMP to obtain compound 24;

[0019] Preparation of compounds 25 and 26: Compound 4 was epoxidized in the presence of m-chloroperoxybenzoic acid (m-CPBA) to prepare compound 25. Compound 25 was then oxidized / rearranged by Des Martin oxidant to prepare compounds 25 and 26.

[0020] This invention also provides the application of a novel compound 1-26, namely eucalyptane-guaiacane sesquiterpene dimer 1-26, in the preparation of an anti-hepatocellular carcinoma drug.

[0021] The present invention also provides a pharmaceutical composition comprising at least one of the compounds of formula (I) 1-26 described above and a pharmaceutically acceptable carrier.

[0022] The application of the pharmaceutical composition in the preparation of anti-liver cancer drugs.

[0023] The method for preparing the pharmaceutical composition involves first obtaining compounds 1-26 using the method for preparing compounds 1-26, and then adding at least one of them to a pharmaceutically acceptable carrier.

[0024] The pharmaceutical composition provided by this invention comprises at least one of compounds 1-26 described above and a pharmaceutically acceptable carrier. In this invention, the pharmaceutically acceptable carrier is preferably a solid, semi-solid, or liquid diluent, filler, or pharmaceutical excipient. This invention does not impose any particular limitation on the pharmaceutically acceptable carrier; any pharmaceutically acceptable carrier well-known in the art that is non-toxic and inert to humans and animals may be selected.

[0025] The present invention does not impose any particular limitation on the preparation method of the pharmaceutical composition. At least one of compounds 1-26 can be directly mixed with a pharmaceutically acceptable carrier. The present invention does not impose any particular limitation on the mixing process. Any process well known in the art that can obtain the pharmaceutical composition can be selected.

[0026] This invention provides the application of the pharmaceutical composition described above in the preparation of anti-liver cancer drugs. This invention does not specifically limit the method of application; any method well-known in the art can be used.

[0027] In this invention, when the pharmaceutical composition is used to prepare an anti-liver cancer drug, the content of the composition in the drug is preferably 0.1% to 99%; in the pharmaceutical composition, the content of at least one of compounds 1-26 is preferably 0.5% to 90%. The pharmaceutical composition of this invention is preferably used in the form of a dose per unit body weight. In this invention, the prepared drug is preferably administered by both injection (intravenous injection, intramuscular injection) and oral administration.

[0028] Compared with the prior art, the present invention has the following advantages:

[0029] 1. This invention provides a series of novel eucalyptane-guaiacane sesquiterpene dimers 1-26 (compounds 1-26).

[0030] 2. This invention provides a novel method for preparing the new compound 1−26, which uses readily available raw materials, is simple in process, and is easy to operate.

[0031] 3. This invention provides a pharmaceutical composition with novel compound 1-26 as the active ingredient, providing a new drug with good pharmaceutical effects for new anti-liver cancer drugs.

[0032] 4. Compounds 1-26 of this invention exhibit strong inhibitory activity against the proliferation of three liver cancer cell lines (HepG2, SK-Hep-1, and Huh7). 20 compounds showed stronger inhibitory activity against HepG2 than sorafenib, 21 compounds showed stronger inhibitory activity against SK-Hep-1 than sorafenib, and 19 compounds showed stronger inhibitory activity against Huh7 than sorafenib. Compound 10 showed the strongest activity, with an IC50 value of [missing information]. 50 The values ​​were 2.1 μM (HepG2), 3.3 μM (SK-Hep-1), and 2.8 μM (Huh7), respectively.

[0033] 5. The synthesized eucalyptane-guaiacane sesquiterpene dimer 1-26 (compound 1-26) or pharmaceutical compositions containing it can be used as drugs to treat liver cancer-related diseases. Attached Figure Description

[0034] Figure 1 This is a schematic diagram of the structural formula of the eucalyptane-guaiacane sesquiterpene dimer 1-26 (compound 1-26) of the present invention. Detailed Implementation

[0035] To better understand the present invention, the following description, in conjunction with the accompanying drawings, uses embodiments of the present invention to further illustrate its essence, but this description is not intended to limit the scope of the invention.

[0036] Example 1:

[0037] Preparation of compounds 1-6.

[0038]

[0039] Eucalyptane-type sesquiterpenes α-cyclocostunolide, β-cyclocostunolide, γ-cyclocostunolide, santamarin, reynosin, and magnolialide (0.15 mmol, 1.5 equivalents) were dissolved in dichloromethane (2 mL) along with dienes (29 mg, 0.1 mmol, 1.0 equivalents), concentrated under reduced pressure, and then heated to 50°C in an oil bath. oC, react for 48 h. After the reaction was complete, the mixture was dissolved in methanol (5 mL), and excess iodomethane (250 μL, 4 mmol, 40 equivalents) was added. The mixture was stirred at room temperature for 3 h. After evaporating the methanol, the solid was transferred to a separatory funnel, and ethyl acetate (10 mL) and 10% sodium bicarbonate aqueous solution (10 mL) were added. The mixture was shaken until the solid was completely dissolved. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (2 × 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether, 25:75) to finally obtain compounds 1 (33 mg), 2 (35 mg), 3 (30 mg), 4 (32 mg), 5 (33 mg), and 6 (30 mg).

[0040] Compound 1

[0041] Yield: 69% (33 mg)

[0042] Molecular formula: C 30 H 38 O5

[0043] Molecular weight: 478.6290

[0044] HRMS (ESI, m / z): [M + H] + [C 30 H 39 O5] + Calculated value: 479.2792; Measured value: 479.2796.

[0045] 11H NMR (500 MHz, CDCl3) δ 6.12 (d, 1H, J = 3.5 Hz, H-13a), 5.51 (d, 1H, J = 10.0 Hz, H-6), 5.41 (d, 1H, J = 3.5 Hz, H-13b), 5.33 (s, 1H, H-3'), 3.84 (dd, 1H, J = 10.0, 10.0 Hz, H-6'), 3.04 - 3.03 (m, 1H, H-2), 2.84 - 2.79 (m, 1H, H-7), 2.18 - 1.98 (m, 7H, H-7', H-5', H-13'a, H-8a, H-2'a, H-2'b, H-9a), 1.79 - 1.70 (m, 6H, H-8'a, H-8b, H-9b, H-15'), 1.62 - 1.53 (m, 3H, H-8'b, H-9'a, H-13'b), 1.47 - 1.34 (m, 11H, H-1'a, H-1'b, H-3a, H-3b, H-9'b, H-14, H-15), 0.85 (s, 3H, H-14). 13 13C NMR (125 MHz, CDCl3) δ 151.4 (C-1), 42.8 (C-2), 53.3 (C-3), 59.7 (C-4), 144.1 (C-5), 83.5 (C-6), 46.7 (C-7), 23.5 (C-8), 38.5 (C-9), 72.6 (C-10), 140.3 (C-11), 170.2 (C-12), 118.8 (C-13), 27.3 (C-14), 17.2 (C-15), 37.7 (C-1'), 22.9 (C-2'), 122.6 (C-3'), 133.1 (C-4'), 50.7 (C-5'), 79.1 (C-6'), 51.7 (C-7'), 21.4 (C-8'), 39.1 (C-9'), 35.1 (C-10'), 55.7 (C-11'), 181.6 (C-12'), 35.0 (C-13'), 17.1 (C-14'), 23.7 (C-15').

[0046] Compound 2

[0047] Yield: 73% (35 mg)

[0048] Molecular formula: C 30H 38 O5

[0049] Molecular weight: 478.6290

[0050] HRMS (ESI, m / z): [M + Na] + [C 30 H 38 O5Na] + Calculated value 501.2611, measured value 501.2621.

[0051] 1 H NMR (500 MHz, CDCl3) δ 6.10 (d, 1H, J = 3.5 Hz, H-13a), 5.49 (d, 1H, J = 10.0 Hz, H-6), 6.10 (d, 1H, J = 3.5 Hz, H-13b), 4.86 (s, 1H, H-15'a), 4.64 (s, 1H, H-15'b), 3.96 (dd, J = 10.0, 10.0 Hz, 1H, H-6'), 3.03 (d, 1H, J = 3.5 Hz, H-2), 2.84 - 2.78 (m, 1H, H-7), 2.34 (brs, 1H, OH), 2.30 - 2.26 (m, 1H, H-3'a), 2.21 - 2.15 (m, 2H, H-13'a, H-7'), 2.12 - 2.06 (m, 2H, H-5', H-8a), 2.00 - 1.95 (m, 2H, H-9a, H-3'b), 1.80 - 1.75 (m, 1H, H-9b), 1.73 - 1.66 (m, 1H, H-8b), 1.64 - 1.55 (m, 6H, H-2'a, H-2'b, H-8'a, H-8'b, H-13'b, H-1'a), 1.44 - 1.32 (m, 11H, H-14, H-15, H-1'b, H-9'b, H-9'a, H-3a, H-3b), 0.79 (s, H-14'). 13C NMR (125MHz, CDCl3) δ 151.8 (C-1), 42.7 (C-2), 53.4 (C-3), 59.6 (C-4), 143.7 (C-5), 83.4 (C-6), 46.7 (C-7), 23.3 (C-8), 38.4 (C-9), 72.5 (C-10), 140.3 (C-11), 170.3 (C-12), 118.8 (C-13), 27.0 (C-14), 17.3 (C-15), 39.5 (C-1'), 23.0 (C-2'), 36.1 (C-3'), 145.1 (C-4'), 54.5 (C-5'), 76.8 (C-6'), 50.2 (C-7'), 21.4(C-8'), 41.6 (C-9'), 37.9 (C-10'), 56.2 (C-11'), 181.3 (C-12'), 35.0 (C-13'), 17.8 (C-14'), 108.5 (C-15').

[0052] Compound 3

[0053] Yield: 62% (30 mg)

[0054] Molecular formula: C 30 H 38 O5

[0055] Molecular weight: 478.6290

[0056] HRMS (ESI, m / z): [M + Na] + [C 30 H 38 O5Na] + Calculated value: 501.2611; Measured value: 501.2604.

[0057] 11H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.52 (d, 1H, J = 10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 4.54 (d, 1H, H-6'), 3.06 - 3.05 (m, 1H, H-2), 2.84 - 2.79 (m, 1H, H-7), 2.28 - 2.24 (m, 2H, H-7', H-13'a), 2.13 - 2.09 (m, 1H, H-8a), 2.03 - 1.98 (m, 2H, H-3'a, H-9a), 1.90 - 1.86 (m, 1H, H-3'b), 1.83 (s, 3H, H-15), 1.80 - 1.55 (m, 8H, H-8'a, H-9b, H-8b, H-8'b, H-2'a, H-2'b, H-13'b, H-1'a), 1.48 - 1.40 (m, 9H, H-1'b, H-9'a, H-9'b, H-15', H-14'), 1.35 (brs, 2H, H-3a, H-3b), 1.07 (s, 3H, H-14'). 13 13C NMR (125 MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.4 (C-3), 59.9 (C-4), 144.2 (C-5), 83.6 (C-6), 46.8 (C-7), 23.5 (C-8), 38.5 (C-9), 72.5 (C-10), 140.1 (C-11), 170.2 (C-12), 118.7 (C-13), 27.4 (C-14), 17.4 (C-15), 40.8 (C-1'), 18.8 (C-2'), 34.7 (C-3'), 129.6 (C-4'), 127.1 (C-5'), 80.2 (C-6'), 50.8 (C-7'), 22.1 (C-8'), 40.5 (C-9'), 36.5 (C-10'), 55.8 (C-11'), 180.9 (C-12'), 34.8 (C-13'), 25.7 (C-14'), 20.3 (C-15').

[0058] Compound 4

[0059] Yield: 65% (32 mg)

[0060] Molecular formula: C 30 H 38 O6

[0061] Molecular weight: 494.6280

[0062] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 39 O8] − Calculated value: 539.2650; Measured value: 539.2645.

[0063] 1 H NMR (500 MHz, CDCl3) δ 6.13 (d, 1H, J = 3.5 Hz, H-13a), 5.50 (d,1H, J = 10.5 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 5.30 (brs, 1H, H-3'),3.91 (dd, 1H, J = 10.0, 10.0 Hz, H-6'), 3.62 (t, 1H, J = 8.0 Hz, H-1'), 3.04-3.03 (m, 1H, H-2), 2.85-2.79 (m, 1H, H-7), 2.32-2.28 (m, 1H, H-2'), 2.19-2.09(m, 4H, H-8a, H-7', H-5', H-13'a), 2.02-1.89 (m, 3H, H-9a, H-9'a, H-2'b), 1.80-1.77 (m, 2H, H-9b, H-8b), 1.74 (s, 3H, H-15'), 1.69-1.67 (m, 2H, H-9'b,H-8'b), 1.56 (d, 1H, J = 11.5 Hz, H-13'b), 1.41 (s, 3H, H-15), 1.38 (s, 3H,H-14), 1.35 (brs, 2H, H-3a, H-3b), 0.82 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.5 (C-1), 42.7 (C-2), 53.4 (C-3), 59.7 (C-4), 143.9 (C-5), 83.8(C-6), 46.7 (C-7), 23.4 (C-8), 38.4 (C-9), 133.3 (C-4'), 50.6 (C-5'), 78.6 (C-6'), 51.3 (C-7'), 21.0 (C-8'), 34.0 (C-9'), 40.0 (C-10'), 55.5 (C-11'), 181.6 (C-12'), 35.0 (C-13'), 10.9 (C-14'), 23.5 (C-15').

[0064] Compound 5

[0065] Yield: 67% (33 mg)

[0066] Molecular formula: C 30 H 38 O6

[0067] Molecular weight: 494.6280

[0068] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 39 O8] − Calculated value: 539.2650; Measured value: 539.2644.

[0069] 1H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.54 (d,1H, J = 10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 4.95 (s, 1H, H-15'a),4.75 (s, 1H, H-15'b), 4.05 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.52 (dd, 1H, J= 11.5, 4.5 Hz, H-1'), 3.07-3.06 (m, 1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.34-2.30 (m, 1H, H-3'a), 2.33-1.98 (m, 7H, H-3'b, H-5', H-7', H-13'a, H-8a, H-9a,H-9'a), 1.82-1.54 (m, 8H, H-8b, H-9b, H-9'b, H-8'a, H-8'b, H-2'a, H-2'b, H-13'b), 1.44 (s, 3H, H-15), 1.42 (s, 3H, H-14), 1.39-1.35 (m, 2H, H-3a, H-3b),0.79 (s, 3H, H-14'). 13 C NMR (125 MHz, CDCl3) δ 151.2 (C-1), 42.7 (C-2), 53.4(C-3), 59.7 (C-4), 144.1 (C-5), 83.3 (C-6), 46.7 (C-7), 23.3 (C-8), 38.6 (C-9), 72.7 (C-10), 140.1 (C-11), 170.2 (C-12), 118.8 (C-13), 27.3 (C-14), 17.3(C-15), 76.3 (C-1'), 31.3 (C-2'), 33.6 (C-3'), 143.0 (C-4'), 52.5 (C-5'),78.1 (C-6'), 49.8 (C-7'), 21.1 (C-8'), 35.5 (C-9'), 42.1 (C-10'), 56.2 (C-11'), 181.0 (C-12'), 35.0 (C-13'), 11.5 (C-14'), 110.1 (C-15').

[0070] Compound 6

[0071] Yield: 60% (30 mg)

[0072] Molecular formula: C 30 H 38 O6

[0073] Molecular weight: 494.6280

[0074] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 39 O8] − Calculated value: 539.2650; Measured value: 539.2650.

[0075] 1 H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.0 Hz, H-13a), 5.52 (d,1H, J = 10.5 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 4.57 (d, 1H, J = 11.5Hz, H-6'), 3.54 (dd, 1H, J = 10.5, 5.0 Hz, H-1'), 3.06-3.05 (m, 1H, H-2), 2.85-2.79 (m, 1H, H-7), 2.28-1.97 (m, 8H, H-3'a, H-3'b, H-7', H-2'a, H-13'a,H-8a, H-9a, H-9'a), 1.83 (s, 3H, H-15'), 1.82-1.78 (m, 2H, H-8b, H-9b), 1.73-1.68 (m, 4H, H-9'b, H-8'a, H-8'b, H-2'b), 1.57 (d, 1H, J = 12.0 Hz, H-13'b), 1.43 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.38-1.34 (m, 2H, H-3a, H-3b), 1.06 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.5 (C-3), 60.0 (C-4), 144.2 (C-5), 83.5 (C-6), 46.7 (C-7), 23.5 (C-8), 38.5 (C-9),72.5 (C-10), 140.1 (C-11), 170.2 (C-12), 118.9 (C-13), 27.4 (C-14), 17.5 (C-15), 77.3 (C-1'), 27.1 (C-2'), 33.6 (C-3'), 126.7 (C-4'), 128.7 (C-5'), 79.9 (C-6'), 50.3 (C-7'), 21.8 (C-8'), 37.6 (C-9'), 41.4 (C-10'), 55.6 (C-11'), 180.8 (C-12'), 34.8 (C-13'), 18.2 (C-14'), 20.1 (C-15').

[0076] Example 2:

[0077] Preparation of compounds 7-8.

[0078]

[0079] Under argon protection, compound C1 (96 mg, 0.2 mmol, 1.0 equivalent) was dissolved in a mixture of tetrahydrofuran and nitromethane (9:1, 5 mL), and NFSI (315 mg, 1.0 mmol, 5.0 equivalent) was added in an ice-water bath. After reacting at room temperature for 12 hours, the mixture was quenched with water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether, 25:75) to give a mixture of compounds 7 and 8. These compounds were further separated by semi-preparative high-performance liquid chromatography (YMC-Pack ODS-A, 5 µm, 250 × 10 mm) with methanol-water (75:25, 3 mL / min) as the mobile phase, finally yielding compound 7 (20 mg) and compound 8 (56 mg).

[0080] Compound 7

[0081] Yield: 20% (20 mg)

[0082] Molecular formula: C 30 H 37 FO5

[0083] Molecular weight: 496.6194

[0084] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 38 O7F] − Calculated value: 541.2607; Measured value: 541.2598

[0085] 1 H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.52 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 5.31 (m, 1H, H-3'),4.48 (ddd, 1H, J = 50.5, 9.5, 7.5 Hz, H-1'), 3.93 (dd, 1H, J = 10.5, 10.5 Hz,H-6'), 3.06-3.05 (m, 1H, H-2), 2.85-2.80 (m, 1H, H-7), 2.51-2.45 (m, 1H, H-2'a), 2.26-2.09 (m, 5H, H-5', H-2'b, H-7', H-8a, H-13'a), 2.07-1.96 (m, 2H, H-9'a, H-9a), 1.84-1.69 (m, 7H, H-8b, H-9b, H-8'a, H-8'b, H-15'), 1.58-1.56 (m,1H, H-13'b), 1.43 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.40-1.36 (m, 3H, H-9'b,H-3a, H-3b), 0.92 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.3 (C-1), 42.8(C-2), 53.4 (C-3), 59.8 (C-4), 144.3 (C-5), 83.5 (C-6), 46.7 (C-7), 23.5 (C-8), 38.7 (C-9), 72.7 (C-10), 140.2 (C-11), 170.2 (C-12), 118.9 (C-13), 27.5(C-14), 17.2 (C-15), 96.0 (d, J = 179.1 Hz, C-1'), 30.0 (d, J = 21.6 Hz, C-2'), 120.5 (d, J = 12.6 Hz, C-3'), 133.8 (d, J = 2.0 Hz, C-4'), 49.9 (d, J =5.8 Hz, C-5'), 78.0 (d, J = 4.5 Hz, C-6'), 51.3 (C-7'), 21.0 (C-8'), 34.0 (d,J = 3.5 Hz, C-9'), 39.7 (d, J = 16.6 Hz, C-10'), 55.5 (C-11'), 181.1 (C-12'), 35.1 (C-13'), 11.4 (d, J = 1.8 Hz, C-14'), 23.2 (C-15').

[0086] Compound 8

[0087] Yield: 56% (56 mg)

[0088] Molecular formula: C 30 H 37 FO5

[0089] Molecular weight: 496.6194

[0090] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 38 O7F] − Calculated value: 541.2607; Measured value: 541.2619

[0091] 1H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-11a), 5.51 (d,1H, J = 10.5 Hz, H-6), 5.41 (d, 1H, J = 3.0 Hz, H-11b), 4.97 (s, 1H, H-15'a),4.77 (s, 1H, H-15'b), 4.34 (ddd, 1H, J = 48.5, 11.5, 5.0 Hz, H-1'), 4.03 (dd,1H, J = 10.0, 10.0 Hz, H-6'), 3.05-3.04 (m, 1H, H-2), 2.83-2.81 (m, 1H, H-7),2.37-2.33 (m, 1H, H-3'a), 2.22-2.17 (m, 2H, H-7', H-9'a), 2.14-2.04 (m, 4H,H-8a, H-3'b, H-13'a, H-5'), 2.02-1.96 (m, 2H, H-2'a, H-9a), 1.82-1.58 (m, 6H,H-9b, H-8b, H-8'a, H-8'b, H-2'b, H-13'b), 1.48-1.42 (m, 1H, H-9'b), 1.43 (s,3H, H-15), 1.40 (s, 3H, H-14), 1.37-1.34 (m, 2H, H-3a, H-3b), 0.84 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.6 (C-1), 42.8 (C-2), 53.5 (C-3), 59.8 (C-4), 143.9 (C-5), 83.4 (C-6), 46.7 (C-7), 23.4 (C-8), 38.6 (C-9), 72.7 (C-10), 140.3 (C-11), 170.3 (C-12), 118.9 (C-13), 27.3 (C-14), 17.3 (C-15), 98.4 (d,J = 180.3 Hz, C-1'), 28.3 (d, J = 18.7 Hz, C-2'), 32.7 (d, J = 12.9 Hz, C-3'), 142.1 (d, J = 2.0 Hz, C-4'), 51.5 (d, J = 6.1 Hz, C-5'), 76.0 (d, J =4.1 Hz, C-6'), 49.8 (C-7'), 21.1 (C-8'), 35.3 (d, J = 2.1 Hz, C-9'), 41.7 (d,J = 17.1 Hz, C-10'), 56.2 (C-11'), 180.9 (C-12'), 35.1 (C-13'), 11.9 (d, J =2.8 Hz, C-14'), 111.0 (C-15').

[0092] Example 2:

[0093] Preparation of compounds 9-17.

[0094]

[0095] Under argon protection, a solution of compound C2 (96 mg, 0.2 mmol, 1.0 equivalent) dissolved in tetrahydrofuran-water (9:1, 5 mL) was added to NCS, NBS, or NIS (0.2 mmol, 1.0 equivalent) at 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes, then allowed to rise naturally to room temperature and stirred for another 20 hours. After the reaction was complete, the mixture was quenched with sodium thiosulfate solution and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether, 25:75). Using NCS, a mixture of compound 11 (14 mg) with compounds 9 and 10 was given; using NBS, a mixture of compound 14 (14 mg) with compounds 12 and 13 was given; and using NIS, a mixture of compound 17 (12 mg) with compounds 15 and 16 was given. The mixture was further purified by high performance liquid chromatography (Agilent, Eclipse XDB-C18, 5 µm, 9.4 mm × 250 mm) with methanol-water (85:15, 3 mL / min) as the mobile phase, and finally obtained compounds 9 (19 mg), 10 (41 mg), 12 (20 mg), 13 (40 mg), 15 (19 mg) and 16 (42 mg).

[0096] Compound 9

[0097] Yield: 19% (19 mg)

[0098] Molecular formula: C 30 H 37 ClO5

[0099] Molecular weight: 513.0710

[0100] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 38 O7Cl] − Calculated value: 557.2312; Measured value: 557.2318

[0101] 1H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.53 (d,1H, J =10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 5.32 (brs, 1H, H-3'),4.06-4.03 (m, 1H, H-1'), 3.91 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.07 (brs,1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.63-2.57 (m, 1H, H-2'a), 2.38-2.30 (m, 2H,H-2'b, H-5'), 2.19-2.05 (m, 4H, H-9'a, H-13'a, H-8a, H-7'), 2.02-1.96 (m, 1H,H-9a), 1.85-1.81 (m, 2H, H-8a, H-8b), 1.78 (s, 3H, H-15'), 1.71-1.68 (m, 2H,H-8'a, H-8'b), 1.58-1.56 (m, 1H, 13'b), 1.44 (s, 3H, H-15), 1.42 (s, 3H, H-14), 1.38-1.30 (m, 3H, H-3a, H-3b, H-9'b), 0.94 (s, 3H, H-14'). 13C NMR (125MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.3 (C-3), 59.7 (C-4), 144.2 (C-5), 83.4 (C-6), 46.6 (C-7), 23.4 (C-8), 38.6 (C-9), 72.7 (C-10), 140.1 (C-11), 170.1 (C-12), 118.8 (C-13), 27.4 (C-14), 17.1 (C-15), 67.8 (C-1'), 34.9 (C-2'), 121.5 (C-3'), 133.7 (C-4'), 51.7 (C-5'), 78.3 (C-6'), 51.2 (C-7'), 20.9 (C-8'), 35.7 (C-9'), 40.4 (C-10'), 55.4 (C-11'), 181.0 (C-12'), 35.0 (C-13'), 11.3 (C-14'), 23.3 (C-15').

[0102] Compound 10

[0103] Yield: 40% (41 mg)

[0104] Molecular formula: C 30 H 37 ClO5

[0105] Molecular weight: 513.0710

[0106] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 38 O7Cl] − Calculated value: 557.2312, Measured value: 557.2292

[0107] 1H NMR (500 MHz, CDCl3) δ 6.13 (d, 1H, J = 3.5 Hz, H-13a), 5.50 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.0 Hz, H-13b), 4.96 (s, 1H, H-15'a),4.76 (s, 1H, H-15'b), 4.03 (dd, 1H, J = 10. 5, 10.5 Hz, H-6'), 3.92 (dd, 1H,J = 12.5, 4.5 Hz, H-1'), 3.06-3.05 (m, 1H, H-2), 2.86-2.80 (m, 1H, H-7),2.36-2.33 (m, 1H, H-3'a), 2.23-2.08 (m, 7H, H-7', H-5', H-9'a, H-2'a, H-3'b,H-13'a, H-8a), 2.00-1.96 (m, 1H, H-9a), 1.92-1.85 (m, 1H, H-2'b), 1.81-1.57(m, 5H, H-8b, H-9b, H-8'a, H-8'b, H-13'b), 1.42 (s, 3H, H-15), 1.40 (s, 3H,H-14), 1.37-1.34 (m, 3H, H-3a, H-3b, H-9'b), 0.89 (s, 3H, H-14'). 13C NMR (125MHz, CDCl3) δ 151.6 (C-1), 42.7 (C-2), 53.5 (C-3), 59.7 (C-4), 143.8 (C-5), 83.3 (C-6), 46.7 (C-7), 23.4 (C-8), 38.6 (C-9), 142.0 (C-4'), 54.1 (C-5'), 76.3 (C-6'), 49.8 (C-7'), 21.1(C-8'), 36.9 (C-9'), 42.9 (C-10'), 56.2 (C-11'), 180.9 (C-12'), 35.0 (C-13'),12.2 (C-14'), 110.8 (C-15').

[0108] Compound 11

[0109] Yield: 13% (14 mg)

[0110] Molecular formula: C 30 H 39 O6Cl

[0111] Molecular weight: 531.0860

[0112] HRMS (ESI, m / z): [M + H] + [C 30 H 40 O6Cl] + Calculated value: 531.2508; Measured value: 531.2547

[0113] 1H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.50 (d,1H, J = 10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 4.11 (dd, 1H, J =11.0, 11.0 Hz, H-6'), 4.00-3.97 (m, 1H, H-1'), 3.26 (s, 1H, OH), 3.08-3.07(m, 1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.30-2.25 (m, 1H, H-7'), 2.20-2.06 (m,5H, H-13'a, H-2'a, H-3'a, H-8a, H-2'b), 2.01-1.97 (m, 1H, H-9a), 1.87-1.79(m, 4H, H-5', H-9'a, H-9b, H-8b), 1.71-1.67 (m, 2H, H-3'b, H-8'a), 1.64-1.58(m, 2H, H-13'b, H-8'b), 1.44 (s, 3H, H-15), 1.43 (s, 1H, H-14), 1.42-1.39 (m,1H, H-3a), 1.33 (s, 3H, H-15'), 1.33-1.31(m, 1H, H-3b), 1.08 (s, 3H, H-14'). 13 C NMR (125 MHz, CDCl3) δ 151.6 (C-1), 42.8 (C-2), 53.5 (C-3), 59.9 (C-4), 144.0 (C-5), 83.2 (C-6), 46.7 (C-7), 23.4 (C-8), 38.7 (C-9), 72.9 (C-10), 140.0 (C-11), 170.1 (C-12), 119.1 (C-13), 27.4 (C-14), 17.4 (C-15), 64.9 (C-1'), 31.5 (C-2'), 41.3 (C-3'), 71.0 (C-4'), 57.3 (C-5'), 78.3 (C-6'), 50.8 (C-7'), 21.4 (C-8'), 41.2 (C-9'), 41.7 (C-10'), 55.5 (C-11'), 179.8 (C-12'), 35.2 (C-13'), 16.0 (C-14'), 24.3 (C-15').

[0114] Compound 12

[0115] Yield: 18% (20 mg)

[0116] Molecular formula: C 30 H 37 O5Br

[0117] Molecular weight: 557.5250

[0118] HRMS (ESI, m / z): [M + H] + [C 30 H 38 O5Br] + Calculated value: 557.1897; Measured value: 557.1863

[0119] 1H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.52 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 5.25 (brs, 1H, H-3'),4.25-4.22 (m, 1H, H-1'), 3.89 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.06-3.05(m, 1H, H-2), 2.85-2.79 (m, 1H, H-7), 2.72-2.66 (m, 1H, H-2'a), 2.60-2.53 (m,1H, H-2'b), 2.37 (d, 1H, J = 10.5 Hz, H-5'), 2.18-2.11 (m, 4H, H-7', H-9'a,H-13'a, H-8a), 2.01-1.96 (m, 1H, H-9a), 1.85-1.78 (m, 2H, H-8b, H-9b), 1.77(s, 3H, H-15'), 1.70-1.67 (m, 2H, H-8'a, H-8'b), 1.58-1.56 (m, 1H, H-13'b),1.42 (s, 3H, H-15), 1.10 (s, 3H, H-14), 1.39-1.28 (m, 3H, H-3a, H-3b, H-9'b). 13C NMR (125 MHz, CDCl3) δ 151.3 (C-1), 42.8 (C-2), 53.4 (C-3), 59.8 (C-4), 144.2 (C-5), 83.4 (C-6), 46.7 (C-7), 23.5 (C-8), 38.6 (C-9), 72.7 (C-10),140.2 (C-11), 170.2 (C-12), 119.0 (C-13), 27.4 (C-14), 17.2 (C-15), 63.2 (C-1'), 36.2 (C-2'), 122.3 (C-3'), 133.8 (C-4'), 51.5 (C-5'), 78.6 (C-6'), 51.4(C-7'), 21.1 (C-8'), 37.7 (C-9'), 40.2 (C-10'), 55.5 (C-11'), 181.1 (C-12'), 35.0 (C-13'), 12.3 (C-14'), 23.4 (C-15').

[0120] Compound 13

[0121] Yield: 36% (40 mg)

[0122] Molecular formula: C 30 H 37 O5Br

[0123] Molecular weight: 557.5250

[0124] HRMS (ESI, m / z): [M + H] + [C 30 H 38 O5Br] + Calculated value: 557.1897, Measured value: 557.1866

[0125] 11H NMR (500 MHz, CDCl3) δ 6.11 (d, 1H, J = 3.5 Hz, H-13a), 5.47 (d, 1H, J = 10.0 Hz, H-6), 5.40 (d, 1H, J = 3.5 Hz, H-13b), 4.94 (s, 1H, H-15'a), 4.73 (s, 1H, H-15'b), 4.13 - 4.10 (m, 1H, H-1'), 4.01 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.04 - 3.03 (m, 1H, H-2), 2.83 - 2.78 (m, 1H, H-7), 2.30 - 2.27 (m, 1H, H-3'a), 2.22 - 1.94 (m, 9H, H-3'b, H-5', H-7', H-8a, H-13'a, H-2'a, H-2'b, H-9'a, H-9a), 1.79 - 1.56 (m, 5H, H-9b, H-8b, H-13'b, H-8'a, H-8'b), 1.39 (s, 3H, H-15), 1.37 (s, 3H, H-14), 1.36 - 1.30 (m, 3H, H-9'b, H-3a, H-3b), 0.91 (s, 3H, H-14'). 13 13C NMR (125 MHz, CDCl3) δ 151.8 (C-1), 42.7 (C-2), 53.5 (C-3), 59.6 (C-4), 143.5 (C-5), 83.2 (C-6), 46.6 (C-7), 23.3 (C-8), 38.4 (C-9), 72.5 (C-10), 140.3 (C-11), 170.2 (C-12), 118.9 (C-13), 27.1 (C-14), 17.3 (C-15), 65.6 (C-1'), 34.6 (C-2'), 36.6 (C-3'), 142.0 (C-4'), 54.2 (C-5'), 76.5 (C-6'), 49.9 (C-7'), 21.1 (C-8'), 38.4 (C-9'), 42.8 (C-10'), 56.2 (C-11'), 180.9 (C-12'), 34.9 (C-13'), 13.3 (C-14'), 110.7 (C-15').

[0126] Compound 14

[0127] Yield: 12% (14 mg)

[0128] Molecular formula: C 30 H 39 O6Br

[0129] Molecular weight: 575.5400

[0130] HRMS (ESI, m / z): [M + Na] + [C 30 H 39 O6BrNa] + Calculated value: 597.1822; Measured value: 597.1830

[0131] 1 H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.49 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J =3.0 Hz, H-13b), 4.10 (dd, 1H, J =11.0, 11.0 Hz, H-6'), 3.98 (dd, 1H, J = 12.0, 4.0 Hz, H-1'), 3.29 (brs, 1H,OH), 3.07-3.06 (m, 1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.29-2.24 (m, 1H, H-7'),2.19-2.03 (m, 5H, H-2'a, H-2'b, H-3'a, H-13'a, H-8a), 2.00-1.95 (m, 1H, H-9a), 1.86-1.76 (m, 4H, H-5', H-9'a, H-8b, H-9b), 1.71-1.58 (m, 4H, H-8'a, H-8'b, H-3'b, H-13'b), 1.43 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.39-1.38 (m,1H, H-3a), 1.36-1.30 (m, 5H, H-15', H-3b, H-9'b), 1.08 (s, 3H, H-14'). 13C NMR(125 MHz, CDCl3) δ 151.6 (C-1), 42.8 (C-2), 53.5 (C-3), 59.9 (C-4), 143.8 (C-5), 83.1 (C-6), 46.7 (C-7), 23.4 (C-8), 38.7 (C-9), 72.8 (C-10), 140.2 (C-11), 170.1 (C-12), 119.0 (C-13), 27.3 (C-14), 17.4 (C-15), 64.9 (C-1'), 31.4(C-2'), 41.3 (C-3'), 71.0 (C-4'), 57.2 (C-5'), 78.2 (C-6'), 50.8 (C-7'), 21.4(C-8'), 41.1 (C-9'), 41.6 (C-10'), 55.5 (C-11'), 179.8 (C-12'), 35.1 (C-13'), 16.0 (C-14'), 24.3 (C-15').

[0132] Compound 15

[0133] Yield: 16% (19 mg)

[0134] Molecular formula: C 30 H 37 O5I

[0135] Molecular weight: 604.5255

[0136] HRMS (ESI, m / z): [M + H] + [C 30 H 38 O5I] + Calculated value: 605.1758; Measured value: 605.1772

[0137] 1H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.53 (d,1H, J = 10.5 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 5.13 (brs, 1H, H-3'),4.43 (dd, 1H, J = 9.0, 9.0 Hz, H-1'), 3.89 (dd, 1H, J = 11.0, 11.0 Hz, H-6'),3.07-3.06 (m, 1H, H-2), 2.83-2.80 (m, 3H, H-7, H-2'a, H-2'b), 2.46 (d, 1H, J= 11.0 Hz, H-5'), 2.19-2.11 (m, 3H, H-13'a, H-8a, H-7'), 2.04-1.97 (m, 2H, H-13'a, H-8a), 1.84-1.77 (m, 2H, H-9'a, H-9a), 1.77 (s, 3H, H-15'), 1.69-1.66(m, 2H, H-8'a, H-8'b), 1.58-1.55 (m, 1H, H-13'b), 1.43 (s, 3H, H-15), 1.42(s, 3H, H-14), 1.38-1.25 (m, 3H, H-3a, H-3b, H-9'b), 0.95 (s, 3H, H-14'). 13CNMR (125 MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.3 (C-3), 59.8 (C-4), 144.2(C-5), 83.4 (C-6), 46.6 (C-7), 23.4 (C-8), 38.9 (C-9), 72.7 (C-10), 140.0 (C-11), 170.1 (C-12), 118.9 (C-13), 27.4 (C-14), 17.1 (C-15), 46.5 (C-1'), 38.6(C-2'), 123.2 (C-3'), 133.9 (C-4'), 50.1 (C-5'), 78.9 (C-6'), 51.7 (C-7'), 21.2 (C-8'), 41.4 (C-9'), 39.5 (C-10'), 55.5 (C-11'), 181.1 (C-12'), 34.7 (C-13'), 13.9 (C-14'), 23.3 (C-15').

[0138] Compound 16

[0139] Yield: 35% (42 mg)

[0140] Molecular formula: C 30 H 37 O5I

[0141] Molecular weight: 604.5255

[0142] HRMS (ESI, m / z): [M + Na] + [C 30 H 37 O5INa] + Calculated value: 627.1578; Measured value: 627.1589

[0143] 1H NMR (500 MHz, CDCl3) δ 6.12 (d, 1H, J = 3.5 Hz, H-13a), 5.48 (d,1H, J = 10.0 Hz, H-6), 5.41 (d, 1H, J = 3.5 Hz, H-13b), 4.93 (s, 1H, H-15'a),4.73 (s, 1H, H-15b), 4.33 (dd, 1H, J = 13.0, 4.5 Hz, H-1'), 4.00 (dd, 1H, J =10.5, 10.5 Hz, H-6'), 3.05-3.04 (m, 1H, H-2), 2.84-2.79 (m, 1H, H-7), 2.41-2.37 (m, 1H, H-3'a), 2.30-2.26 (m, 2H, H-3'b, H-5'), 2.20-2.10 (m, 5H, H-7',H-2'a, H-2'b, H-13'a, H-8a), 2.02-1.95 (m, 2H, H-9a, H-9'a), 1.81-1.71 (m,2H, H-8b, H-9b), 1.66-1.56 (m, 3H, H-8'a, H-8'b, H-13'b), 1.40 (s, 3H, H-15),1.37 (s, 3H, H-14), 1.35-1.29 (m, 3H, H-9'b, H-3a, H-3b), 0.92 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.6 (C-1), 42.7 (C-2), 53.5 (C-3), 59.7 (C-4), 143.7 (C-5), 83.2 (C-6), 46.7 (C-7), 23.4 (C-8), 38.5 (C-9), 72.6 (C-10),140.3 (C-11), 170.2 (C-12), 118.9 (C-13), 27.6 (C-14), 17.4 (C-15), 49.2 (C-1'), 38.5 (C-2'), 37.2 (C-3'), 142.2 (C-4'), 53.5 (C-5'), 76.8 (C-6'), 50.1(C-7'), 21.2 (C-8'), 41.5 (C-9'), 42.4 (C-10'), 56.2 (C-11'), 181.0 (C-12'), 35.0 (C-13'), 15.4 (C-14'), 110.7 (C-15').

[0144] Compound 17

[0145] Yield: 10% (12 mg)

[0146] Molecular formula: C 30 H 39 O6I

[0147] Molecular weight: 622.5405

[0148] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 40 O8I] − Calculated value: 667.1773, Measured value: 667.1753

[0149] 1H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.49 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 4.16-4.13 (m, 1H, H-1'), 4.08 (dd, J = 11.0, 11.0 Hz, 1H, H-6'), 3.07-3.06 (m, 1H, H-2), 2.85-2.80 (m, 1H, H-7), 2.36-2.32 (m, 1H, H-2'a), 2.28-2.22 (m, 2H, H-7', H-2'b),2.18-2.10 (m, 2H, H-13'a, H-8a), 1.97-1.90 (m, 3H, H-9a, H-5', H-9'a), 1.83-1.78 (m, 2H, H-8b, H-9b), 1.67-1.57 (m, 5H, H-13'b, H-3'a, H-3'b, H-8'a, H-8'b), 1.42 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.39-1.38 (m, 1H, H-3a), 1.33-1.29 (m, 5H, H-15', H-3b, H-13'b), 1.08 (s, 3H, H-14'). 13 C NMR (125 MHz,CDCl3) δ 151.6 (C-1), 42.8 (C-2), 53.5 (C-3), 59.8 (C-4), 143.8 (C-5), 83.1(C-6), 47.6 (C-7), 23.3 (C-8), 38.6 (C-9), 72.8 (C-10), 140.1 (C-11), 170.1(C-12), 119.0 (C-13), 27.2 (C-14), 17.4 (C-15), 46.7 (C-1'), 34.0 (C-2'),43.8 (C-3'), 71.3 (C-4'), 56.2 (C-5'), 78.7 (C-6'), 51.0 (C-7'), 21.2 (C-8'),43.0 (C-9'), 41.2 (C-10'), 55.5 (C-11'), 179.9 (C-12'), 35.1 (C-13'), 18.5(C-14'), 24.3 (C-15').

[0150] Example 4:

[0151] Preparation of compounds 18 and 19.

[0152]

[0153] Under argon protection, compound C2 (96 mg, 0.2 mmol, 1.0 equivalent) was dissolved in tetrahydrofuran (5 mL), cooled to 0 °C, and then a tetrahydrofuran solution (0.25 mL) of dimethyl(methylthio)thiotetrafluoroborate (DMTSF, 98 mg, 0.4 mmol, 2.0 equivalent) was slowly added dropwise. The reaction system was slowly heated to room temperature and stirred for 12 hours. The reaction solution was quenched with water, extracted with ethyl acetate, and the combined organic phases were washed successively with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether, 25:75) to give compounds 18 (34 mg) and 19 (19 mg), respectively.

[0154] Compound 18

[0155] Yield: 32% (34 mg)

[0156] Molecular formula: C 31 H 40 O5S

[0157] Molecular weight: 524.7160

[0158] HRMS (ESI, m / z): [M + HCOO] − [C 32 H 41 O7S] − Calculated value: 569.2578; Measured value: 569.2565

[0159] 1H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.53 (d,1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 4.94 (s, 1H, H-15'a),4.72 (s, 1H, H-15'b), 4.02 (dd, 1H, J = 11.0, 11.0 Hz, H-6'), 3.06-3.05 (m,1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.49 (dd, 1H, J = 13.0, 3.5 Hz, H-1'),2.39-2.31 (m, 2H, H-3'a, H-9'a), 2.21-2.18 (m, 2H, H-7', H-13'a), 2.16-2.04(m, 7H, SMe, H-5', H-3'b, H-2'a, H-8a), 2.01-1.96 (m, 1H, H-9a), 1.81-1.77(m, 2H, H-8b, H-9b), 1.67-1.64 (m, 3H, H-2'b, H-8'a, H-8'b), 1.59-1.57 (m,1H, H-13'b), 1.43 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.38-1.35 (s, 3H, H-9'b,H-3a, H-3b), 0.82 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.3 (C-1), 42.7(C-2), 53.4 (C-3), 59.7 (C-4), 144.0 (C-5), 83.3 (C-6), 46.7 (C-7), 23.3 (C-8), 38.6 (C-9), 72.7 (C-10), 140.2 (C-11), 170.2 (C-12), 118.8 (C-13), 27.3(C-14), 17.3 (C-15), 59.1 (C-1'), 31.1 (C-2'), 36.2 (C-3'), 143.1 (C-4'),55.2 (C-5'), 76.4 (C-6'), 49.9 (C-7'), 21.2 (C-8'), 37.6 (C-9'), 42.6 (C-10'), 56.2 (C-11'), 181.0 (C-12'), 35.0 (C-13'), 13.5 (C-14'), 109.7 (C-15'),16.2 (SMe).

[0160] Compound 19

[0161] Yield: 18% (19 mg)

[0162] Molecular formula: C 31 H 40 O5S

[0163] Molecular weight: 524.7160

[0164] HRMS (ESI, m / z): [M + HCOO] − [C 32 H 41 O7S] − Calculated value: 569.2578; Measured value: 569.2577

[0165] 1H NMR (500 MHz, CDCl3) δ 6.14 (d, 1H, J = 3.5 Hz, H-13a), 5.43 (d,1H, J = 3.5 Hz, H-13b), 5.40 (d, 1H, J = 10.0 Hz, H-6), 4.96 (s, 1H, H-15'a),4.75 (s, 1H, H-15'b), 4.01 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.18-3.17 (m,1H, H-2), 2.97-2.91 (m, 1H, H-7), 2.50 (dd, 1H, J = 13.0, 4.0 Hz, H-1'),2.37-2.27 (m, 3H, H-3'a, H-8a, H-9'a), 2.22-2.07 (m, 8H, H-7', H-13'a, SMe,H-5', H-2'a, H-3'b), 1.88-1.85 (m, 2H, H-9a, H-9b), 1.71-1.60 (m, 4H, H-2'b,H-8b, H-8'a, H-8'b), 1.47-1.33 (m, 10H, H-15, H-3a, H-3b, H-9'b, H-13'b, H-14), 0.83 (s, 3H, H-14'). 13 C NMR (125 MHz, CDCl3) δ 150.0 (C-1), 42.2 (C-2),53.7 (C-3), 59.8 (C-4), 144.2 (C-5), 84.2 (C-6), 47.6 (C-7), 24.2 (C-8), 39.8(C-9), 73.3(C-10), 139.9 (C-11), 170.3 (C-12), 118.7 (C-13), 27.7 (C-14),17.4 (C-15), 59.1 (C-1'), 31.1 (C-2'), 36.2 (C-3'), 143.1 (C-4'), 55.2 (C-5'), 76.3 (C-6'), 49.9 (C-7'), 21.2 (C-8'), 37.6 (C-9'), 42.6 (C-10'), 56.0(C-11'), 180.2 (C-12'), 34.6 (C-13'), 13.5 (C-14'), 109.8 (C-15'), 16.2(SMe)。

[0166] Example 5:

[0167] Preparation of compounds 20 and 21.

[0168]

[0169] Under argon protection, phenylselenyl chloride (PhSeCl, 19 mg, 0.1 mmol, 1.0 equivalent) was dissolved in anhydrous dichloromethane, and silver trifluoromethanesulfonate (AgOTf, 26 mg, 0.1 mmol, 1.0 equivalent) was added at 0 °C. After stirring at 0 °C for 10 min, pyridine (40 µL, 0.5 mmol, 5.0 equivalent) and compound C2 (48 mg, 0.1 mmol, 1.0 equivalent) in dichloromethane (2 mL) were added sequentially. After reacting for 1 hour, the reaction was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was initially purified by silica gel column chromatography (acetone-petroleum ether, 25:75) to obtain a mixture of compounds 20 and 21. The mixture was further separated by high performance liquid chromatography (Agilent, Eclipse XDB-C18, 5 µm, 9.4 mm × 250 mm) with methanol-water (85:15, 3 mL / min) as the mobile phase to obtain compounds 20 (23 mg) and 21 (10 mg).

[0170] Compound 20

[0171] Yield: 36% (23 mg)

[0172] Molecular formula: C 36 H 42 O5Se

[0173] Molecular weight: 633.6870

[0174] HRMS (ESI, m / z): [M + HCOO] − [C 37 H 43 O7Se] − Calculated value: 679.2183; Measured value: 679.2176

[0175] 1H NMR (500 MHz, CDCl3) δ 7.59-7.57 (m, 2H, H-2'', H-6''), 7.29-7.27(m, 3H, H-4'', H-3'', H-5''), 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.54 (d, 1H, J= 10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 4.97 (s, 1H, H-15'a), 4.73(s, 1H, H-15'b), 4.02 (dd, 1H, J = 10.5, 10.5 Hz, H-6'), 3.18 (dd, 1H, J =13.0, 4.0 Hz, H-1'), 3.07-3.06 (m, 1H, H-2), 2.86-2.81 (m, 1H, H-7), 2.38-2.30 (m, 2H, H-9'a, H-3'a), 2.22-1.92 (m, 8H, H-13'a, H-5', H-7', H-2'a, H-8a, H-9a, H-3'b, H-2'b), 1.82-1.76 (m, 2H, H-8b, H-9b), 1.66-1.58 (m, 3H, H-13'b, H-8'a, H-8'b), 1.42-1.33 (m, 9H, H-14, H-15, H-9'b, H-3a, H-3b), 0.92(s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.3 (C-1), 42.7 (C-2), 53.3 (C-3), 59.7 (C-4), 144.1 (C-5), 83.3 (C-6), 46.7 (C-7), 23.3 (C-8), 38.6 (C-9), 143.0 (C-4'), 55.2 (C-5'), 76.5 (C-6'), 49.9 (C-7'), 21.2 (C-8'), 38.9 (C-9'), 42.8 (C-10'), 56.2 (C-11'), 180.9 (C-12'), 35.0 (C-13'), 14.7 (C-14'), 109.8 (C-15'), 129.9 (C-1''), 134.9 (C-2'', C-6''), 129.2 (C-3'', C-5''), 127.6 (C-4'').

[0176] Compound 21

[0177] Yield: 16% (10 mg)

[0178] Molecular formula: C 36 H 42 O5Se

[0179] Molecular weight: 633.6870

[0180] HRMS (ESI, m / z): [M + HCOO] − [C 37 H 43 O7Se] − Calculated value: 679.2183; Measured value: 679.2177

[0181] 1H NMR (500 MHz, CDCl3) δ 7.58-7.56 (m, 2H, H-2'', H-6''), 7.28-7.26(m, 3H, H-3'', H-4', H-5''), 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.54 (d, 1H, J= 10.0 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 5.31-5.30 (m, 1H, H-3'),3.88 (dd, 1H, J = 11.0, 11.0 Hz, H-6'), 3.41-3.38 (m, 1H, H-1'), 3.07-3.05(m, 1H, H-2), 2.84-2.81 (m, 1H, H-7), 2.52-2.48 (m, 1H, H-2'a), 2.38-2.28 (m,2H, H-5', H-2'b), 2.25-1.96 (m, 5H, H-9'a, H-8b, H-13'a, H-7', H-9a), 1.82-1.79 (m, 1H, H-9b), 1.76 (s, 3H, H-15'), 1.70-1.64 (m, 2H, H-8'a, H-8'b),1.57-1.54 (m, 1H, H-13'b), 1.42 (s, 3H, H-14), 1.41 (s, 3H, H-15), 1.37-1.33(m, 2H, H-3a, H-3b), 1.28-1.24 (m, 1H, H-9'b), 0.96 (s, 3H, H-14'). 13C NMR(125 MHz, CDCl3) δ 151.0 (C-1), 42.7 (C-2), 53.2 (C-3), 59.8 (C-4), 144.3 (C-5), 83.4 (C-6), 46.6 (C-7), 23.4 (C-8), 38.6 (C-9), 133.5 (C-4'), 52.0 (C-5'), 78.8 (C-6'), 51.2 (C-7'), 21.2 (C-8'), 37.9 (C-9'), 40.2 (C-10'), 55.5 (C-11'), 181.2 (C-12'), 35.0 (C-13'), 13.6 (C-14'), 23.5 (C-15'),129.8 (C-1''), 134.8 (C-2'', C-6''), 129.1(C-3'', C-5''), 127.5 (C-4'').

[0182] Example 6:

[0183] Preparation of compounds 22 and 23.

[0184]

[0185] At room temperature, 4-dimethylaminopyridine (DMAP, 12 mg, 0.1 mmol, 2.0 equivalence) and either acetic anhydride (10 µL, 0.1 mmol, 2.0 equivalence) or benzoic anhydride (23 mg, 0.1 mmol, 2.0 equivalence) were added sequentially to a 2.0 mL solution of compound 4 (25 mg, 0.05 mmol) in dichloromethane, and the reaction was allowed to proceed for 4 hours. After the reaction was completed, the mixture was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic phases were washed successively with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. The crude product was purified by silica gel column chromatography (acetone-petroleum ether, 25:75) to give compound 22 (26 mg) or compound 23 (28 mg), respectively.

[0186] Compound 22

[0187] Yield: 97% (26 mg)

[0188] Molecular formula: C32 H 40 O7

[0189] Molecular weight: 536.6650

[0190] HRMS (ESI, m / z): [M + HCOO] − [C 33 H 41 O9] − Calculated value: 581.2756; Measured value: 581.2744

[0191] 1 H NMR (500 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.53 (d,1H, J = 10.5 Hz, H-6), 5.43 (d, 1H, J = 3.5 Hz, H-13b), 5.33 (brs, 1H, H-3'),4.88 (m, 1H, H-1'), 3.91 (dd, 1H, J = 10.0, 10.0 Hz, H-6'), 3.06-3.05 (m, 1H,H-2), 2.85-2.81 (m, 1H, H-7), 2.45-2.42 (m, 1H, H-2'a), 2.34 (d, 1H, J = 11.0Hz, H-5'), 2.18-2.10 (m, 3H, H-8a, H-7', H-13'a), 2.05 (s, 3H, OAc), 2.02-1.98 (m, 2H, H-9a, H-9'a), 1.85-1.72 (m, 7H, H-2'b, H-9b, H-8b, H-15'), 1.68-1.66 (m, 2H, H-8'b, H-8'b), 1.58-1.55 (m, 1H, H-13'b), 1.42 (s, 3H, H-15), 1.41 (s, 3H, H-14), 1.39-1.33 (m, 3H, H-3a, H-3b, H-9'b), 0.94 (s, 3H, H-14'). 13C NMR (125 MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.3 (C-3), 59.7 (C-4), 144.2 (C-5), 83.4 (C-6), 46.6 (C-7), 23.4 (C-8), 38.5 (C-9), 72.7 (C-10),140.0 (C-11), 170.2 (C-12), 118.9 (C-13), 27.3 (C-14), 17.1 (C-15), 76.7 (C-1'), 29.3 (C-2'), 120.9 (C-3'), 133.4 (C-4'), 50.4 (C-5'), 78.0 (C-6'), 51.1(C-7'), 21.0 (C-8'), 33.9 (C-9'), 38.8 (C-10'), 55.4 (C-11'), 181.1 (C-12'), 34.9 (C-13'), 12.1 (C-14'), 23.3 (C-15'), 170.7 (OAc), 21.2 (OAc).

[0192] Compound 23

[0193] Yield: 94% (28 mg)

[0194] Molecular formula: C 37 H 42 O7

[0195] Molecular weight: 598.7360

[0196] HRMS (ESI, m / z): [M + HCOO] − [C 38 H 43 O9] − Calculated value: 643.2913; Measured value: 643.2898

[0197] 1H NMR (500 MHz, CDCl3) δ 8.03 (d, 2H, J = 8.0 Hz, H-2'', H-6''), 7.58(dd, 1H, J = 8.0, 8.0 Hz, H-3''), 7.46-6.15 (dd, 2H, J = 8.0, 8.0 Hz, H-3'',H-5''), 6.15 (d, 1H, J = 3.5 Hz, H-13b), 5.54 (d, 1H, J = 10.5 Hz, H-6), 5.43(d, 1H, J = 3.5 Hz, H-13b), 5.39 (brs, 1H, H-3'), 5.17-5.14 (m, 1H, H-1'),3.96 (dd, 1H, J = 10.0, 10.0 Hz, H-6'), 3.07-3.06 (m, 1H, H-2), 2.86-2.80 (m,1H, H-7), 2.58 (d, 1H, J = 19.0 Hz, H-2'a), 2.43 (d, 1H, J = 11.0 Hz, H-5'),2.21-2.10 (m, 4H, H-8a, H-7', H-13'a, H-9'a), 2.01-1.98 (m, 1H, H-9a), 1.89-1.72 (m, 7H, H-2'b, H-9b, H-8b, H-15'), 1.67-1.65 (m, 2H, H-8'b, H-8'b),1.60-1.57 (m, 1H, H-13'b), 1.46-1.45 (m, 1H, H-9'b), 1.42 (s, 3H, H-15), 1.42(s, 3H, H-14), 1.38-1.32 (m, 2H, H-3a, H-3b), 1.09 (s, 3H, H-14'). 13C NMR(125 MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.3 (C-3), 59.7 (C-4), 144.2 (C-5), 83.4 (C-6), 46.6 (C-7), 23.4 (C-8), 38.6 (C-9), 72.7 (C-10), 140.1 (C-11), 170.2 (C-12), 118.9 (C-13), 27.3 (C-14), 17.1 (C-15), 77.2 (C-1'), 29.5(C-2'), 120.9 (C-3'), 133.5 (C-4'), 50.4 (C-5'), 78.0 (C-6'), 51.1 (C-7'), 20.9 (C-8'), 34.1 (C-9'), 39.3 (C-10'), 55.4 (C-11'), 181.1 (C-12'), 35.0 (C-13'), 12.4 (C-14'), 23.3 (C-15'), 130.3 (C-1''), 129.6 (C-2'', C-6''), 128.5(C-3'', C-5''), 133.1 (C-4''), 166.0 (C-7'').

[0198] Example 7:

[0199] Preparation of compound 24.

[0200]

[0201] At 0 °C, Dess-Martin reagent (DMP, 39 mg, 0.09 mmol, 1.5 equivalence) was added to 1 mL of dichloromethane containing compound 4 (30 mg, 0.06 mmol, 1.0 equivalence), and the mixture was slowly heated to room temperature and reacted for 2 hours. After the reaction was complete, the mixture was quenched with sodium thiosulfate solution and extracted with dichloromethane. The organic phases were combined, washed successively with brine, dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether, 20:80) to give compound 24.

[0202] Compound 24

[0203] Yield: 86% (26 mg)

[0204] Molecular formula: C 30 H 36 O6

[0205] Molecular weight: 492.2160

[0206] HRMS (ESI, m / z): [M + HCOO] − [C 31 H 37 O8] − Calculated value: 537.2494; Measured value: 537.2484

[0207] 1 H NMR (400 MHz, CDCl3) δ 6.16 (d, 1H, J = 3.6 Hz, H-13a), 5.54 (d,1H, J = 10.0 Hz, H-6), 5.44 (d, 1H, J = 3.5 Hz, H-13b), 5.30 (s, 1H, H-3'),4.08 (dd, 1H, J = 10.0, 10.0 Hz, H-6'), 3.08-3.07 (m, 1H, H-2), 3.03-2.83 (m,3H, H-2'a, H-2'b, H-7), 2.62 (d, 1H, J =10.0 Hz, H-5'), 2.21-2.11 (m, 2H, H-7', H-13'a), 1.92 (s, 3H, H-15'), 2.04-1.98 (m, 2H, H-9a, H-8a), 1.83-1.61 (m, 6H, H-8'a, H-8b, H-9b, H-8'b, H-9'a, H-9'b), 1.44 (s, 3H, H-15), 1.44 (s,3H, H-14), 1.40-1.34 (m, 2H, H-3a, H-3b), 1.10 (s, 3H, H-14). 13C NMR (100MHz, CDCl3) δ 151.1 (C-1), 42.7 (C-2), 53.3 (C-3), 59.8 (C-4), 144.2 (C-5), 83.3 (C-6), 46.6 (C-7), 23.4 (C-8), 38.6 (C-9), 72.7 (C-10), 140.0 (C-11), 170.1 (C-12), 118.9 (C-13), 27.4 (C-14), 17.2 (C-15), 212.3 (C-1'), 31.2 (C-2'), 119.3 (C-3'), 135.6 (C-4'), 48.4 (C-5'), 77.4 (C-6'), 50.4 (C-7'), 20.4(C-8'), 38.6 (C-9'), 47.4 (C-10'), 55.7 (C-11'), 180.7 (C-12'), 35.0 (C-13'), 16.7 (C-14'), 22.1 (C-15').

[0208] Example 8:

[0209] Preparation of compounds 25 and 26.

[0210]

[0211] At 0 °C, 85% m-chloroperoxybenzoic acid (m-CPBA, 12 mg, 0.06 mmol, 1.2 equivalence) was added to a dichloromethane (1 mL) solution of compound 4 (25 mg, 0.05 mmol, 1.0 equivalence) and reacted at room temperature for 5 h. The reaction was then quenched with 10% sodium thiosulfate solution and extracted with dichloromethane. The combined organic phases were washed successively with brine, dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (acetone-petroleum ether, 30:70) to give compound 25.

[0212] Compound 25

[0213] Yield: 90% (23mg)

[0214] Molecular formula: C 30 H 38 O7

[0215] Molecular weight: 510.6270

[0216] HRMS (ESI, m / z): [M + HCOO] − [C31 H 39 O9] − Calculated value 555.2600, measured value 555.2581

[0217] 1 1H NMR (400 MHz, CDCl3) δ 6.15 (d, 1H, J = 3.5 Hz, H-13a), 5.54 (d, 1H, J = 10.0 Hz, H-6), 5.44 (d, 1H, J = 3.5 Hz, H-13b), 3.91 (dd, 1H, J = 11.0, 11.0 Hz, H-6'), 3.46 - 3.42 (m, 1H, H-1'), 3.08 - 3.07 (m, 1H, H-2), 2.85 - 2.80 (m, 1H, H-7), 3.00 (d, 1H, J = 3.0 Hz, H-3'), 2.87 - 2.82 (m, 1H, H-7), 2.44 (dd, 1H, J = 15.0 Hz, 6.0 Hz, H-2'a), 2.20 - 2.12 (m, 3H, H-5', H-8a, H-13'a), 2.02 - 1.96 (m, 1H, H-9a), 1.93 - 1.89 (m, 1H, 9'a), 1.83 - 1.58 (m, 7H, H-2'b, H-8'a, H-8'b, H-9b, H-8b, H-13'b), 1.43 (s, 3H, H-15'), 1.43 (s, 3H, H-15), 1.42 (s, 3H, H-14), 1.41 - 1.43 (m, 2H, H-3a, H-3b), 1.25 - 1.19 (m, 1H, H-9'b), 0.89 (s, 3H, H-14'). 13C NMR (100 MHz, CDCl3) δ 151.4 (C-1), 42.8 (C-2), 53.4 (C-3), 59.8 (C-4), 144.0 (C-5), 83.4 (C-6), 46.7 (C-7), 23.4 (C-8), 38.6 (C-9), 72.8 (C-10), 140.1 (C-11), 170.4 (C-12), 119.1 (C-13), 27.2 (C-14), 17.3 (C-15), 73.6 (C-1'), 30.9 (C-2'), 152.1 (C-3'), 57.4 (C-4'), 50.8 (C-5'), 77.8 (C-6'), 52.2 (C-7'), 20.9 (C-8'), 34.0 (C-9'), 39.4 (C-10'), 55.7 (C-11'), 180.9 (C-12'), 35.0 (C-13'), 11.7 (C-14'), 21.6 (C-15').

[0218]

[0219] Compound 25 (46 mg, 0.09 mmol, 1.0 equivalent) was dissolved in dichloromethane (1 mL) at 0 °C, followed by the addition of sodium bicarbonate (230 mg, 0.27 mmol, 3.0 equivalent) and Dess-Martin reagent (DMP, 57 mg, 0.14 mmol, 1.5 equivalent). The mixture was slowly heated to room temperature and stirred for 15 minutes. The reaction was then quenched with sodium thiosulfate solution and extracted with dichloromethane. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was mixed with silica gel (2 g), heated at 60 °C for 4 hours, and then purified by silica gel column chromatography (acetone-petroleum ether, 40:60) to obtain compound 26.

[0220] Compound 26

[0221] Yield: 87% (40 mg)

[0222] Molecular formula: C 30 H 36 O7

[0223] Molecular weight: 508.6110

[0224] HRMS (ESI, m / z): [M + HCOO] − [C 31 H37 O9] − Calculated value 553.2443, measured value 553.2437

[0225] 1 1H NMR (400 MHz, CDCl3) δ 6.58 (d, 1H, J = 10.5 Hz, H-3'), 6.13 (d, 1H, J = 3.5 Hz, H-13a), 5.87 (d, 1H, J = 10.5 Hz, H-2'), 5.49 (d, 1H, J = 10.0 Hz, H-6), 5.42 (d, 1H, J = 3.5 Hz, H-13b), 4.14 (dd, 1H, J = 11.0, 11.0 Hz, H-6'), 3.14 (s, 1H, OH), 3.07 - 3.06 (m, 1H, H-2), 2.85 - 2.80 (m, 1H, H-7), 2.42 (d, 1H, J = 11.0 Hz, H-5'), 2.25 - 2.11 (m, 3H, H-7', H-13'a, H-9'a), 2.06 - 1.96 (m, 2H, H-9a, H-8a), 1.81 - 1.75 (m, 3H, H-9b, H-8b, H-9'b), 1.66 - 1.60 (m, 3H, H-8'a, H-8'b, H-13'b), 1.50 (s, 3H, H-15'), 1.42 (s, 3H, H-15), 1.40 (s, 3H, H-14), 1.40 - 1.32 (m, 2H, H-3a, H-3b), 1.16 (s, 3H, H-14). 13C NMR (100MHz, CDCl3) δ 151.6 (C-1), 42.7 (C-2), 53.5 (C-3), 59.8 (C-4), 143.6 (C-5), 83.1 (C-6), 46.6 (C-7), 23.3 (C-8), 38.5 (C-9), 72.7 (C-10), 140.0 (C-11), 170.2 (C-12), 119.2 (C-13), 27.4 (C-14), 17.4 (C-15), 201.7 (C-1'), 125.6 (C-2'), 152.1 (C-3'), 70.1 (C-4'), 54.6 (C-5'), 76.8 (C-6'), 50.0 (C-7'), 20.9(C-8'), 33.7 (C-9'), 45.6 (C-10'), 55.6 (C-11'), 180.0 (C-12'), 35.1 (C-13'), 19.6 (C-14'), 23.9 (C-15').

[0226] Example 9:

[0227] To evaluate the cytotoxic activity of guaiacol-eucalyptane sesquiterpene dimer 1-26 against hepatocellular carcinoma cell lines.

[0228] 1. Materials and Methods

[0229] 1.1 Materials

[0230] Hepatocellular carcinoma cell lines (HepG2, Huh7, and SK-Hep-1) were purchased from Shanghai Jining Biotechnology Co., Ltd.; culture medium (Dulbecco's Modified Eagle Medium, DMEM) was purchased from Thermo Fisher Scientific (Suzhou, China); serum (fetal bovine serum, FBS) was purchased from Life Technologies (NY, USA); and RPMI-1640 was purchased from ThermoFisher Biochemical Products (Beijing, China).

[0231] 1.2 Instruments

[0232] Flex Station 3 benchtop multi-functional microplate reader (Bio-RAD 680, USA); analytical balance (AG135, Metler Toledo, China); incubator (DHP-9082, Shanghai).

[0233] 1.3 Experimental Procedure

[0234] 1) Collect liver cancer cells in the logarithmic growth phase, discard the old culture medium, wash twice with PBS, and discard the PBS;

[0235] 2) Digest the cells with 0.25% trypsin. When the cell outlines are observed to darken and become rounded under a microscope, quickly remove the trypsin.

[0236] 3) Stop digestion with DMEM complete medium containing 10% FBS and resuspend the cells. Take 10 μL of the cell suspension, count the cells using a cell counter, and adjust the cell concentration to 1 × 10⁻⁶ cells with culture medium. 4 / mL, seeded into 96-well plates, add 100 μL of cell suspension to each well, and incubate in a 37 ℃, 5% CO2 incubator for 24 h to allow the cells to adhere;

[0237] 4) Remove the culture medium, add the diluted sample to the plate, 100 μL per well, set up 3 replicates for each concentration, and continue to incubate in the incubator for 48 h;

[0238] 5) Remove the culture medium, add the prepared MTT solution (1 mg / mL), add 100 μL to each well, and incubate in an incubator for 4 h;

[0239] 6) Remove the MTT solution, add 100 μL of DMSO to each well, and incubate in an incubator for 10 min;

[0240] 7) Measure the absorbance at 490 nm using a microplate reader. Calculate the cell inhibition rate using the formula: Inhibition rate = (Negative - Experimental group) / (Negative - Blank group) × 100%. Calculate the IC50 using the statistical software GraphPadprism 5. 50 The experiment was repeated 3 times.

[0241] 2. Results

[0242] The cytotoxic activities of compounds 1-26 against three hepatocellular carcinoma cell lines (HepG2, Huh7, and SK-Hep-1) are shown in Table 1. Most compounds exhibited some inhibitory activity against all three cell lines. Twenty compounds showed stronger inhibitory activity against HepG2 than sorafenib, 21 compounds showed stronger inhibitory activity against SK-Hep-1 than sorafenib, and 19 compounds showed stronger inhibitory activity against Huh7 than sorafenib. Compound 10 showed the best activity, with an IC50 value of [missing information - likely a specific value]. 50The values ​​were 2.1, 2.8, and 3.3 μM, respectively, which were superior to the positive control drug sorafenib.

[0243] Compounds 1–6 exhibited 1.9 to 4.8 times higher activity than their corresponding monomers, IC50 50 The values ​​ranged from 6.2 to 18.3 μM. Among them, compounds 4–6 with a hydrophilic group attached at the C-1' position showed lower activity than compounds 1–3, suggesting that the hydrophilic structural unit at this position may have an adverse effect on the activity.

[0244] Halogen-containing derivatives (compounds 7–17) exhibited strong anti-hepatocellular carcinoma activity, IC50. 50 Values ​​ranged from 2.1 to 11.1 μM. The activity varied significantly with the type of halogen: chlorinated and iodine-substituted compounds (9–10, 15–16) exhibited the strongest activity, with inhibitory power 2.4–6.1 times that of sorafenib; brominated derivatives (compounds 12–13) showed moderate activity; and fluorinated compounds 7 and 8 showed activity comparable to sorafenib. Notably, the position of the unsaturated bond in the eucalyptolactone fragment had no significant effect on activity, while hydroxylation of the double bond led to a slight decrease in activity (compare 11 with 9–10, 14 with 12–13, and 17 with 15–16).

[0245] The IC50 values ​​of phenylselenoyl-substituted compounds 18–19 for three cell lines (HepG2, SK-Hep-1, and Huh7) were [not specified]. 50 The values ​​were 8.7 / 8.4 μM, 8.1 / 9.5 μM, and 8.4 / 9.3 μM, respectively, showing slightly better activity than sorafenib. Although the sulfur-containing compounds 20 and 21 have different configurations at the C-6 position, they exhibit similar biological activities, indicating that the stereoconfiguration of the hydroxyl group at this position has no significant effect on the activity.

[0246] The derivatives 22–23 obtained by acylation of compound 4 showed better activity than the parent compound, suggesting that the introduction of an ester group at the C-1' position helps to enhance the anti-hepatocellular carcinoma activity. However, the introduction of an epoxy structure at the C3'-C4' position (compared to 4) resulted in a significant decrease in activity; the introduction of a hydroxyl group at the C4' position (26) also weakened the anti-hepatocellular carcinoma efficacy.

[0247] Table 1. Inhibitory activity of compound 1-26 against three hepatocellular carcinoma cell lines.

[0248]

[0249] 3. Conclusion

[0250] The above results indicate that the novel guaiacol-eudemine sesquiterpene dimer 1−36 is cytotoxic to three liver cancer cell lines (HepG2, Huh7, and SK-Hep-1) and can be used as a drug to treat liver cancer-related diseases.

[0251] Formulation Examples 1-7:

[0252] In the following formulation examples, conventional reagents were selected and formulations were prepared according to existing conventional methods. This application example only demonstrates that at least one of the compounds 1-26 described in this invention can be prepared into different formulations, and no specific limitations are made on the specific reagents and operations:

[0253] 1. Dissolve at least one of compounds 1-26 in DMSO, add water for injection according to conventional methods, filter, fill and sterilize to prepare an injection solution with a concentration of 0.5-5 mg / mL.

[0254] 2. Dissolve at least one of compounds 1-26 in DMSO, then dissolve it in sterile water for injection, stir until dissolved, filter using a sterile suction funnel, then filter aseptically, dispense into ampoules, freeze-dry at low temperature, and then aseptically seal to obtain a powder for injection.

[0255] 3. Add at least one of compounds 1-26 to the excipient at a mass ratio of 9:1 to prepare a powder.

[0256] 4. Add at least one of compounds 1-26 to the excipient at a mass ratio of 5:1, and granulate and compress the mixture into tablets.

[0257] 5. Prepare an oral liquid from at least one of compounds 1-26 using conventional oral liquid preparation methods.

[0258] 6. Add at least one of compounds 1-26 to the excipient at a mass ratio of 5:1 to make capsules.

[0259] 7. Add at least one of compounds 1-26 to the excipient at a mass ratio of 5:1 to prepare granules.

[0260] As can be seen from the above embodiments, the present invention provides a guaiacol-eudemine sesquiterpene dimer, its preparation method and application, and a pharmaceutical composition and its application. The novel guaiacol-eudemine sesquiterpene dimer 1-26 (compound 1-26) provided by the present invention exhibits different degrees of cytotoxic activity against liver cancer cells, can be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition, and can be used to prepare an anti-liver cancer drug.

[0261] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Compound 1-26, as shown in the following structural formula, is the eucalyptane-guaiacane sesquiterpene dimer 1-26.

2. The method for preparing compounds 1-26 as shown in claim 1, characterized in that, This method includes preparing compounds 1-6 by reacting guaiac diene C1 with eucalyptane-type sesquiterpenes α-cyclocostunolide, β-cyclocostunolide, γ-cyclocostunolide, santamarin, reynosin, and magnolialide via Diels-Alder reaction / demethylamineization; or preparing a magnolane-guaiac diemer from guaiac lactone as a raw material and further undergoing intramolecular cyclization to obtain compounds 7-21; and preparing compounds 22-26 by further esterification, oxidation, and epoxidation of compound 4.

3. The method for preparing compounds 1-26 as shown in claim 2, characterized in that, The preparation method mainly includes the following key reaction steps: Preparation of compounds 1-6: Eucalyptane-guaiacane sesquiterpene dimers 1-6 were prepared by reacting guaiacane-type dienes with eucalyptane-type sesquiterpenes α-cyclocostunolide, β-cyclocostunolide, γ-cyclocostunolide, santamarin, reynosin, and magnolialide under appropriate conditions via Diels-Alder reaction / dedimethylamine removal. The appropriate conditions for the Diels-Alder reaction were that the dienes reacted with diephiles under solvent-free conditions with heating. Preparation of compounds 7-8: Using tetrahydrofuran and nitromethane as solvents, compound C2 reacted with N-fluorobisbenzenesulfonamide (NFSI) at 0°C. o Compounds 7 and 8 were prepared by reacting at room temperature (C); Preparation of compounds 9-17: Using tetrahydrofuran and water as solvents, compound C2 reacts with N-chlorosuccinimide (NCS), N-bromosuccinimide (NCS), or N-iodosuccinimide (NIS) at 0°C. o Compounds 9-17 were prepared by reacting at room temperature (C). Preparation of compounds 18 and 19: Compound C2 was reacted with dimethyl(methylthio)sulfonium tetrafluoroborate (DMTSF) at 0°C. o Compounds 18 and 19 were prepared by reacting at room temperature (C); Preparation of compounds 20 and 21: Compound C2 was reacted with phenylselenotrifluoromethane sulfonate (PhSeOTf) in dichloromethane as a solvent at 0°C. o Compounds 20 and 21 were prepared by reaction under C. Preparation of compounds 22 and 23: Compound 4 was reacted with acetic anhydride or benzoic anhydride in the presence of 4-dimethylaminopyridine (DMAP) to prepare compounds 22 and 23; Preparation of compound 24: Compound 4 was prepared by oxidation reaction in the presence of Dess-Martin periodinane DMP to obtain compound 24; Preparation of compounds 25 and 26: Compound 4 was epoxidized in the presence of m-chloroperoxybenzoic acid (m-CPBA) to prepare compound 25. Compound 25 was then oxidized / rearranged by Des Martin oxidant to prepare compounds 25 and 26.

4. The use of compounds 1-26, as shown in the structural formula of claim 1, in the preparation of anti-hepatocellular carcinoma drugs.

5. A pharmaceutical composition comprising at least one of compounds 1-26 of the structural formula of claim 1 and a pharmaceutically acceptable carrier.

6. Use of the pharmaceutical composition of claim 5 in the preparation of an anti-liver cancer drug.

7. The method for preparing the pharmaceutical composition according to claim 5, characterized in that, Compound 1-26 is first prepared by the method of claim 2 or 3, and at least one of them is added to a pharmaceutically acceptable carrier.

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