Polystichic acid derivatives containing 1,2,3-triazole groups, processes for their preparation and uses thereof

Abstract

The application discloses a powder back fern acid derivative containing a 1,2,3-triazole group and a preparation method and application thereof, and belongs to the field of pharmaceutical chemical industry. The powder back fern acid is used as a substrate, first reacts with propargylamine under the action of a condensing agent to obtain an intermediate N-propargyl powder back fern amide, then undergoes a click reaction with an azide compound to synthesize a series of powder back fern acid derivatives containing a 1,2,3-triazole. The powder back fern acid derivatives obtained are tested for cytotoxicity on five common cancer cell lines. The results show that most of the compounds exhibit obvious anti-proliferation activity on the five cancer cell lines, and meanwhile, some of the compounds can reach more than half of the inhibition rate on the five cancer cell lines at a concentration of 10 muM. The synthesis process is simple and fast, and the synthesis efficiency is high. The application is another effective way for synthesizing new compounds by modifying the structure of the powder back fern acid after amination. The powder back fern acid derivatives prepared by synthesis are expected to be applied in the field of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemical industry, and relates to the preparation and application of a compound, specifically, to a fern acid derivative containing a 1,2,3-triazole group, its preparation method and application. Background Technology

[0002] *Aleuritopteris argentea* (Gmel.) Fée) is a plant originally belonging to the genus *Aleuritopteris* in the family Pteridaceae, but is now merged into the genus *Aleuritopteris* in the family Pteridaceae. It gets its name from the silvery-white powdery substance that often covers its back. Its dried stem resembles copper wire, hence its common name "copper wire grass." It is also known as "menstrual regulating grass" because of its blood-activating and menstrual-regulating properties, treating menstrual irregularities, amenorrhea, and abdominal pain. It is used as a folk medicine among ethnic minorities in my country, such as the Mongolians and Tibetans.

[0003] *Pteris vittata* contains various natural secondary metabolites, such as flavonoids, chlorogenic acid, polysaccharides, and terpenoids. In 1962, Hiroyuki Ueda et al. first isolated a hemispherane-type diterpenoid, pteridoside, from it, and determined its basic configuration using chemical derivation (Ageta, H et al. Proc. Symp. Nat. Org. Compd. 1962, 6, 136). Ha Jing et al. also isolated two new diterpenoids from *Pteris vittata* (Li, J.-C. et al. Phytochem. Lett. 2017, 20, 22). Pteridoside and its analogues have also been isolated from the essential oil of *Aquilaria sinensis* and *Aquilaria agallocha* (Afolayan, M. et al. Med. Chem. Res. 2018, 27, 2325-2330).

[0004] Previous pharmacological studies on ferrous sulfate were limited. Chinese invention patent publication number 109810017A discloses the application of a *Tetrapanax papyriferus* extract containing ferrous sulfate in the preparation of anticancer drugs. This extract significantly inhibited angiogenesis in mouse sarcomas and suppressed tumor growth. Previous research by the inventors revealed that modifying the 15-carboxyl group of ferrous sulfate to synthesize ferrous sulfate amide derivatives significantly enhanced its anticancer activity. The synthesized new compound stimulated the release of cytochrome c from mitochondria, cleaved and activated caspase-3 and caspase-9, and led to the cleavage of PARP-1, ultimately inducing apoptosis in HeLa cells through the mitochondrial-mediated endogenous apoptosis pathway.

[0005] 1,2,3-Triazole, as one of the most important heterocyclic compounds, can form various non-covalent interactions with different biological targets, such as hydrophobic interactions, hydrogen bonds, and van der Waals forces. Compounds derivatized with 1,2,3-triazole can produce hybrid molecules with better biological activity. Compounds with 1,2,3-triazole groups exhibit a variety of biological activities, such as antibacterial, antimalarial, antifungal, antiviral, antituberculosis, and antitumor activities. Many natural products can achieve better biological activity after being combined with 1,2,3-triazole groups. However, there is currently no research on the hybridization of fern fiber acid and 1,2,3-triazole groups. Considering that the 1,2,3-triazole group can also be regarded as a bioisosteric amide bond, and that fern fiber acid amide derivatives have good antitumor activity, this invention modifies the structure of fern fiber acid to introduce a 1,2,3-triazole pharmacophore, aiming to discover new anticancer active molecules.

[0006] Click reactions are an important method for preparing 1,2,3-triazole compounds. Copper(I)-catalyzed reactions of azides and terminal alkynes to form 1,2,3-triazoles exhibit high reliability, complete specificity, and good biocompatibility, allowing for the creation of new compound libraries for pharmacological activity screening. Based on this, we employed click reaction conditions, using copper(I) formed from the interaction of copper sulfate and sodium ascorbate as a catalyst, to obtain a series of 1,2,3-triazole derivatives of ferrihydric acid via click reactions. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the prior art and provide a ferrihydric acid derivative containing a 1,2,3-triazole group, its preparation method, and its applications. This invention uses ferrihydric acid isolated from *Pteris vittata* as a substrate, first synthesizing the intermediate N-allylferrihydric acid, and then, under copper ion catalysis, interacting with azide compounds via a click reaction to generate a series of target products. After product separation and structural identification, the antitumor activity of the target products is studied.

[0008] The objective of this invention can be achieved through the following technical solutions:

[0009] The structural formula of the fern acid derivative is as follows:

[0010]

[0011] Compound 3a: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0012]

[0013] Compound 3b: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-o-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0014]

[0015] Compound 3c: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-m-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0016]

[0017] Compound 3d: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-p-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0018]

[0019] Compound 3e: (E)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0020]

[0021] Compound 3f: (E)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0022]

[0023] Compound 3g: (E)-N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0024]

[0025] Compound 3h: (E)-N-((1-(2,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0026]

[0027] Compound 3i: (E)-N-((1-(3,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0028]

[0029] Compound 3j: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0030]

[0031] Compound 3k: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0032]

[0033] Compound 3l: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0034]

[0035] Compound 3m: (E)-N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0036]

[0037] Compound 3n: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphth-1-yl)-3-methyl-N-((1-(naphth-1-yl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide;

[0038]

[0039] Compound 3o: (E)-N-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0040]

[0041] Compound 3p: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide;

[0042]

[0043] Compound 3q: (E)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0044]

[0045] Compound 3r: (E)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0046]

[0047] Compound 3S: (E)-N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0048]

[0049] Compound 3t: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide;

[0050]

[0051] Compound 3u: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphth-1-yl)-3-methyl-N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide;

[0052]

[0053] Compound 3v: (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide;

[0054]

[0055] Compound 3w: (E)-N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide;

[0056]

[0057] Compound 3x: (4-((4-(((E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpentan-2-amido)methyl)-1H-1,2,3-triazol-1-yl)methyl)phenyl)boronic acid.

[0058] The preparation method and synthetic route of the fern acid derivative are as follows:

[0059]

[0060] Wherein, R is phenyl, substituted phenyl, α-naphthyl, benzyl or substituted benzyl.

[0061] The specific steps are as follows:

[0062] a. Dissolve the powder of fern acid (1) in dichloromethane, then add diisopropylethylamine, N,N,N',N'-tetramethylurea hexafluorophosphate and propargylamine in sequence. Stir at room temperature until the reaction is complete, quench the reaction with water, extract, wash, dry the organic phase, and evaporate to dryness. The crude product obtained is separated and purified by silica gel column chromatography to obtain the intermediate product N-propargyl fern acid amide (2).

[0063] b. Dissolve the N-propargyl ferrugin (2) and organic azide obtained in step a in a mixed solution of N,N-dimethylformamide and water. Then add sodium ascorbate and anhydrous copper sulfate in sequence. Stir the reaction system at 60°C. After the reaction is complete, extract, wash, dry the organic phase, and evaporate to dryness. The crude product is separated and purified by silica gel column chromatography to obtain the target compounds (3a-3x).

[0064] The beneficial effects of this invention are:

[0065] 1. The synthesis process is simple and convenient, with high synthesis efficiency, and the purified compound has high purity;

[0066] 2. In the screening of antitumor activity, most compounds showed significant antitumor activity, with some compounds exhibiting particularly pronounced antitumor effects, such as compounds 3b, 3n, 3o, 3q, 3r, 3s, and 3u. Studies were conducted on human non-small cell lung cancer cells (A549), human lung cancer cells (PC-9), human breast cancer cell line (MCF-7), human prostate cancer cells (PC-3), and human cervical cancer cells (HeLa). Most of these compounds showed significant IC50 responses against these five common cancer cell lines. 50 The values ​​were all around 10 μM. Compound 3n showed an IC50 value for A549 and PC-9 cells. 50 The concentrations were 4.0 μM and 7.23 μM, respectively; the IC50 values ​​of compound 3s for MCF-7 cells, PC-3 cells, and HeLa cells were [not specified]. 50The values ​​were 2.12 μM, 5.74 μM and 7.58 μM, respectively. Compared with the other 5 compounds, these two compounds had relatively low cytotoxicity to normal human hepatocytes (HL-7702) and high selectivity between normal hepatocytes and cancer cells. Therefore, it can be determined that 3n and 3s are the two compounds with better anti-tumor effects.

[0067] 3. The fern acid derivative of the present invention has the potential to be developed and applied in the field of anti-tumor drugs. Detailed Implementation

[0068] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0069] 1. Experimental steps for derivative synthesis

[0070] Example 1

[0071]

[0072] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-decahydronaphthyl-1-yl)-3-methyl-N-((prop-2-yn)-1-yl)pent-2-enamide (compound 2):

[0073] Dissolve 960 mg of fern acid (1) powder in 20 mL of dichloromethane. Then, add 585 mg of diisopropylethylamine, 1.36 g of N,N,N',N'-tetramethylurea hexafluorophosphate, and 247 mg of propargylamine sequentially. Stir at room temperature and monitor the reaction by TLC. After the reaction is complete, quench the reaction with water, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, and finally evaporate the ethyl acetate to dryness. The crude product obtained after evaporation is separated and purified by silica gel column chromatography to obtain the intermediate product N-propargyl fern acid amide (2).

[0074] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ5.54 (t, J=5.9Hz, 1H, NH), 5.50 (q, J=1.3Hz, 1H, 14-H), 4.85 (t, J=1.6Hz, 1H, 17-H), 4.50 (d, J=1.7Hz, 1H, 17-H), 4.08 (dd, J=5.2, 2.6Hz, 2H, aC H2), 3.25 (dd, J=11.8, 4.4Hz, 1H, 3-H), 2.40 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.28–2.22(m, 1H, 12-H), 2.22(t, J=2.6Hz, 1H, cH), 2.15(d, J=1.3Hz, 3H, 16-C H3 ), 1.92 (m, J=15.9, 7.8Hz, 2H, 7-H, 12-H), 1.80–1.44 (m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.42–1.34(m, 1H, 6-H), 1.15(td, J=13.2, 3.7Hz, 1H, 1-H), 1.07(dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99(s, 3H, 18-C H3 ), 0.77 (s, 3H, 19-C) H3 ), 0.68 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ166.62(C-15), 156.69(C-13), 147.89(C-8), 117.10(C-14), 10 6.91(C-17), 80.02(aC), 78.90(C-3), 71.54(cC), 55.91(C-9), 54.70(C-5), 39.70(C -12), 39.54(C-10), 39.28(C-4), 38.28(C-7), 37.19(C-1), 29.04(bC), 28.43(C-18) , 28.04(C-2), 24.15(C-6), 21.78(C-11), 18.58(C-16), 15.54(C-19), 14.65(C-20).

[0075] Yield: 91%, Melting point: 138-140℃, Mass spectrometry data: C 23 H 35 NO2[M+H] + Calculated value: 358.2746, measured value: 358.2736.

[0076] The azide compounds required for the synthesis of derivatives 3a-3n were obtained by referring to the synthesis method of phenyl azide compounds. The specific experimental procedure was as follows: Under ice bath conditions, 2 mL of 6 mol / L hydrochloric acid was slowly added dropwise to 2 mL of an aqueous solution of the corresponding aniline compound (2.0 mmol), while continuously stirring. Then, 1 mL of an aqueous solution of sodium nitrite (207 mg, 3 mmol) was added dropwise to the system, and the reaction was continued under ice bath conditions with stirring for 15 min. Afterward, 1 mL of an aqueous solution of sodium azide (156 mg, 2.4 mmol) was slowly added dropwise to the system, and the reaction was allowed to proceed overnight at room temperature. The reaction system was extracted with ethyl acetate, and the combined organic phases were washed successively with saturated sodium bicarbonate solution and saturated brine, and then dried over anhydrous Na₂SO₄. The solvent was removed under vacuum to obtain the corresponding azide compounds. The crude product obtained in this step had high purity and could be used directly in subsequent reactions without further purification.

[0077] The azide compound required for the synthesis of derivative 3o-3x was obtained by referring to the synthesis method of benzyl azide compounds. The specific experimental procedure was as follows: At room temperature, an aqueous solution of sodium azide (156 mg, 2.4 mmol) was slowly added dropwise to 2 mL of acetone solution containing the corresponding benzyl bromide compound (2.0 mmol), and the reaction was allowed to proceed overnight. The reaction system was extracted with ethyl acetate, and the combined organic phases were washed successively with water and saturated brine, then dried over anhydrous Na₂SO₄. The solvent was removed under vacuum to obtain the corresponding azide compound. Similarly, the crude product obtained in this step has high purity and can be used directly in subsequent reactions without further purification.

[0078] Example 2

[0079]

[0080] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3a):

[0081] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of phenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0082] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.05 (s, 1H, cH), 7.74–7.67 (m, 2H, 2×Ph-H), 7.58–7.34 (m, 3 H, 3×Ph-H), 6.55 (s, 1H, NH), 5.57 (s, 1H, 14-H), 4.83 (s, 1H, 17-H), 4.61 (s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.23 (dd, J=11.6, 4.4Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.27–2.18 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.99–1.82(m, 2H, 7-H, 12-H), 1.79–1.50(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.36 (qd, J=12.8, 4.2Hz, 1H, 6-H), 1.13 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.31(C-15), 156.06(C-13), 147.82(C-8), 145.80(bC), 137.03(Ph-C), 129.8 6(2×Ph-C), 128.97(Ph-C), 120.91(cC), 120.64(2×Ph-C), 117.49(C-14), 106.86(C-17), 78.80(C-3), 55.96(C-9), 54.63(C-5), 39.70(C-10), 39.48(C-4), 39.22(C-7), 38.22(C-12), 37.13(C-1), 34.53( aC), 28.41(C-18), 27.96(C-2), 24.08(C-6), 21.80(C-11), 18.60(C-16), 15.55(C-19), 14.59(C-20).

[0083] Yield: 93%, Melting point: 160-162℃, Mass spectrometry data: C 29H 40 N4O2[M+H] + Calculated value: 477.3224, measured value: 477.3224.

[0084] Example 3

[0085]

[0086] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-o-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3b):

[0087] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of o-methylphenyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0088] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.83 (s, 1H, cH), 7.47–7.31 (m, 4H, 4×Ph-H), 6.44 (s, 1H, NH), 5.58 (s, 1H, 14-H), 4.85 (s, 1H, 17-H), 4.67 (s, 2H, aC H2 ), 4.50 (s, 1H, 17-H), 3.25 (dd, J=11.8, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.6, 4.3, 2.4Hz, 1H, 7-H), 2.31–2.22 (m, 1H, 12-H), 2.21 (s, 3H, Ph-C H3 ), 2.21(s, 3H, 16-C H3 ), 1.99–1.84(m, 2H, 7-H, 12-H), 1.81–1.51(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.38 (qd, J=13.0, 4.7Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.99 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.68 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.30(C-15), 156.33(C-13), 147.83(C-8), 144.38(bC), 136.21(Ph-C), 133.68( Ph-C), 131.76 (Ph-C), 130.36 (Ph-C), 127.10 (Ph-C), 126.02 (Ph-C), 124.53 (Ph-C), 117.41 (C-14), 106. 92(C-17), 78.88(C-3), 55.97(C-9), 54.65(C-5), 39.71(C-10), 39.51(C-4), 39.25(C-7), 38.25(C-12), 37.16(C-1), 34.24(aC), 28.43(C-18), 28.00(C-2), 24.11(C-6), 21.81(C-11), 18.63(C-16), 18.04(Ph- C H3), 15.56(C-19), 14.63(C-20).

[0089] Yield: 25%, Mass spectrometry data: C 30 H 42 N4O2[M+H] + Calculated value: 493.3381, measured value: 493.3380.

[0090] Example 4

[0091]

[0092] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-m-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3c):

[0093] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of m-methylphenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally the ethyl acetate was evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0094] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.00 (s, 1H, cH), 7.56–7.43 (m, 2H, 2×Ph-H), 7.41–7.32 (m, 1H, Ph-H), 7.24–7.19 (m, 1 H, Ph-H), 6.48 (t, J=5.8Hz, 1H, NH), 5.57 (d, J=1.8Hz, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.60 (d, J=5.7Hz, 2H, aC H2 ), 4.48(s, 1H, 17-H), 3.22(dd, J=11.7, 4.4Hz, 1H, 3-H), 2.42(s, 3H, Ph-C H3 ), 2.37 (ddd, J=12.7, 4.3, 2.4Hz, 1H, 7-H), 2.28–2.16 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.96–1.84(m, 2H, 7-H, 12-H), 1.77–1.46(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.36 (qd, J=12.7, 4.0Hz, 1H, 6-H), 1.13 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ 167.28 (C-15), 155.92 (C-13), 147.81 (C-8), 145.75 (bC), 140.05 (Ph-C), 137. 01(Ph-C), 129.61(Ph-C), 129.59(Ph-C), 121.25(Ph-C), 120.81(cC), 117.67(Ph-C), 117.53(C-14) , 106.84(C-17), 78.77(C-3), 55.95(C-9), 54.62(C-5), 39.67(C-10), 39.46(C-4), 39.21(C-7), 38. 21(C-12), 37.12(C-1), 34.60(aC), 28.40(C-18), 27.96(C-2), 24.07(C-6), 21.79(C-11), 21.49(Ph- C H3), 18.59(C-16), 15.54(C-19), 14.57(C-20).

[0095] Yield: 78%, Melting point: 141-143℃, Mass spectrometry data: C 30 H 42 N4O2[M+H] + Calculated value: 491.3381, measured value: 491.3369.

[0096] Example 5

[0097]

[0098] Preparation of E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-p-tolyl-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3d):

[0099] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-methylphenyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0100] The NMR data are as follows: 1H NMR (400MHz, CDCl3) δ7.97 (s, 1H, cH), 7.60–7.53 (m, 2H, 2×Ph-H), 7.31–7.26 (m, 2H, 2×Ph-H ), 6.49 (t, J=5.8Hz, 1H, NH), 5.57 (s, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.59 (d, J=5.8Hz, 2H, aC H2 ), 4.47 (s, 1H, 17-H), 3.22 (dd, J=11.8, 4.4Hz, 1H, 3-H), 2.39 (s, 3H,), 2.37–2.32 (m, 1H, 7-H), 2.28–2.16 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.95–1.84(m, 2H, 7-H, 12-H), 1.77–1.51(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.36 (qd, J=12.8, 3.9Hz, 1H, 6-H), 1.12 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.05 (dd, J=12.6, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.27(C-15), 155.92(C-13), 147.82(C-8), 145.71(bC), 138.98(Ph-C ), 134.80(Ph-C), 130.31(2×Ph-C), 120.73(cC), 120.52(2×Ph-C), 117.54(C-14), 106.84(C-1 7), 78.78(C-3), 55.95(C-9), 54.63(C-5), 39.67(C-10), 39.47(C-4), 39.22(C-7), 38.22(C-1 2), 37.13(C-1), 34.61(aC), 28.40(C-18), 27.97(C-2), 24.08(C-6), 21.79(C-11), 21.20(Ph- C H3), 18.58(C-16), 15.54(C-19), 14.58(C-20).

[0101] Yield: 78%, Melting point: 145-146℃, Mass spectrometry data: C30 H 42 N4O2[M+H] + Calculated value: 491.3381, measured value: 491.3369.

[0102] Example 6

[0103]

[0104] Preparation of (E)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3e):

[0105] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-fluorophenyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0106] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.03 (s, 1H, cH), 7.71–7.62 (m, 2H, 2×Ph-H), 7.24–7.13 (m, 2 H, 2×Ph-H), 6.59 (s, 1H, NH), 5.57 (s, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.60 (s, 2H, aC H2 ), 4.47 (s, 1H, 17-H), 3.23 (dd, J=11.6, 4.3Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.29–2.16 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.98–1.80 (m, 2H, 7-H, 12-H), 1.77–1.46 (m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.36 (qd, J=13.0, 4.3Hz, 1H, 6-H), 1.12 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ 171.15 (C-15), 167.21 (C-13), 162.43 (d, J=249.5Hz, Ph-C), 156.10 (C-8), 147.68 (bC), 133. 14 (d, J=3.1Hz, Ph-C), 122.49 (d, J=8.5Hz, 2×Ph-C), 121.12 (cC), 117.26 (C-14), 116.71 (d, J=23.3Hz, 2×Ph-C), 10 6.71(C-17), 78.65(C-3), 55.81(C-9), 54.48(C-5), 39.57(C-10), 39.33(C-4), 39.08(C-7), 38.07(C-12), 36.99( C-1), 34.33(aC), 28.27(C-18), 27.81(C-2), 23.93(C-6), 21.65(C-11), 18.47(C-16), 15.42(C-19), 14.45(C-20).

[0107] Yield: 93%, Melting point: 135-136℃, Mass spectrometry data: C 29 H 39 FN4O2[M+H] + Calculated value: 495.3130, measured value: 495.3128.

[0108] Example 7

[0109]

[0110] Preparation of (E)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3f):

[0111] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-chlorophenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0112] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.07 (s, 1H, cH), 7.67 (d, J=8.7Hz, 2H, 2×Ph-H), 7.49 (d, J=8.6H z, 2H, 2×Ph-H), 6.44 (s, 1H, NH), 5.56 (s, 1H, 14-H), 4.84 (s, 1H, 17-H), 4.62 (s, 2H, aC H2 ), 4.48(s, 1H, 17-H), 3.24(dd, J=11.6, 4.2Hz, 1H, 3-H), 2.38(ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.30–2.18(m, 1H, 12-H), 2.14(s, 3H, 16-C H3 ), 1.99–1.84(m, 2H, 7-H, 12-H), 1.79–1.46(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=13.0, 4.1Hz, 1H, 6-H), 1.14 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.66 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ167.37(C-15), 156.56(C-13), 147.82(C-8), 145.83(bC), 135.36(Ph-C), 135.0 7(Ph-C), 130.15(2×Ph-C), 121.88(2×Ph-C), 121.26(cC), 117.31(C-14), 106.90(C-17), 78.86(C-3), 55.97(C-9), 54.64(C-5), 39.74(C-10), 39.50(C-4), 39.24(C-7), 38.23(C-12), 37.16(C-1), 34.40( aC), 28.42(C-18), 27.98(C-2), 24.09(C-6), 21.82(C-11), 18.65(C-16), 15.56(C-19), 14.62(C-20).

[0113] Yield: 96%, Melting point: 166-167℃, Mass spectrometry data: C 29 H 39 ClN4O2[M+H] + Calculated value: 511.2834, measured value: 511.2836.

[0114] Example 8

[0115]

[0116] Preparation of (E)-N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3g):

[0117] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-bromophenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0118] The NMR data are as follows: 1H NMR (400MHz, CDCl3) δ8.03 (s, 1H, cH), 7.69–7.54 (m, 4H, 4×Ph-H), 6.56 (t, J= 6.0Hz, 1H, NH), 5.56 (s, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.58 (d, J=4.9Hz, 2H, aC H2 ), 4.47 (s, 1H, 17-H), 3.22 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 3.2, 2.4Hz, 1H, 7-H), 2.27–2.17 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.99–1.81(m, 2H, 7-H, 12-H), 1.75–1.47(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.36 (qd, J=13.0, 4.0Hz, 1H, 6-H), 1.12 (td, J=13.2, 3.6Hz, 1H, 1-H), 1.04 (dd, J=12.5, 2.6Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.19(C-15), 156.04(C-13), 147.67(C-8), 146.05(bC), 135.88(Ph-C), 132.8 3(2×Ph-C), 122.37(Ph-C), 121.81(2×Ph-C), 120.60(cC), 117.27(C-14), 106.69(C-17), 78.63(C-3), 55.82(C-9), 54.48(C-5), 39.57(C-10), 39.33(C-4), 39.07(C-7), 38.07(C-12), 36.99(C-1), 34.40( aC), 28.27(C-18), 27.82(C-2), 23.93(C-6), 21.65(C-11), 18.46(C-16), 15.41(C-19), 14.44(C-20).

[0119] Yield: 87%, Melting point: 187-188℃, Mass spectrometry data: C 29 H 39 BrN4O2[M+H] +Calculated value: 557.2309, measured value: 557.2299.

[0120] Example 9

[0121]

[0122] Preparation of (E)-N-((1-(2,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3h):

[0123] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of 2,4-dichlorophenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0124] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.98 (s, 1H, cH), 7.60–7.46 (m, 2H, 2×Ph-H), 7.41 (dd, J=8.6, 2.2Hz, 1H, Ph-H), 6.50–6.32 (m, 1H, Ph-H), 5.56 (s, 1H, 14-H), 4.83 (s, 1H, 17-H), 4.62 (d, J=5.6Hz, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.23 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 4.2, 2.4Hz, 1H, 7-H), 2.28–2.16 (m, 1H, 12-H), 2.13 (s, 3H, 16), 1.95–1.85 (m, 2H, 7-H, 12-H), 1.78–1.49 (m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.36 (qd, J=12.8, 4.1Hz, 1H, 6-H), 1.13 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-CH3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ171.13(C-15), 167.10(C-13), 155.93(C-8), 147.67(bC), 136.24(Ph-C), 133.44(Ph-C ), 130.56(Ph-C), 129.39(Ph-C), 128.43(Ph-C), 128.21(Ph-C), 124.33(cC), 117.30(C-14), 106.70(C-17), 78.65(C-3), 55.80(C-9), 54.49(C-5), 39.52(C-10), 39.33(C-4), 39.07(C-7), 38.07(C-12), 36.99(C-1), 3 4.40(aC), 28.25(C-18), 27.82(C-2), 23.94(C-6), 21.62(C-11), 18.43(C-16), 15.39(C-19), 14.44(C-20).

[0125] Yield: 64%, Mass spectrometry data: C 29 H 38 Cl2N4O2[M+H] + Calculated value: 545.2445, measured value: 545.2437.

[0126] Example 10

[0127]

[0128] Preparation of (E)-N-((1-(3,4-dichlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3i):

[0129] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of 3,4-dichlorophenyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0130] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.02 (s, 1H, cH), 7.90 (t, J=1.4Hz, 1H, Ph-H), 7.59 (d, J=1.4Hz, 2H, 2×Ph- H), 6.16 (t, J=5.9Hz, 1H, Ph-H), 5.54 (s, 1H, 14-H), 4.85 (s, 1H, 17-H), 4.60 (d, J=7.3Hz, 2H, aC H2 ), 4.49 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.4, 2.5Hz, 1H, 7-H), 2.31–2.19 (m, 1H, 12-H), 2.15 (s, 3H, 16-C H3 ), 1.99–1.85(m, 2H, 7-H, 12-H), 1.78–1.55(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.38 (qd, J=13.0, 4.3Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.99 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ167.29(C-15), 156.55(C-13), 147.85(C-8), 146.42(bC), 136.12(C-Ph), 134.15(C-Ph ), 133.04(C-Ph), 131.59(C-Ph), 122.39(C-Ph), 120.72(cC), 119.50(Ph-C), 117.31(C-14), 106.90(C-17), 78.88(C-3), 55.99(C-9), 54.68(C-5), 39.75(C-10), 39.53(C-4), 39.26(C-7), 38.26(C-12), 37.18(C-1), 3 4.67(aC), 28.43(C-18), 28.02(C-2), 24.12(C-6), 21.84(C-11), 18.63(C-16), 15.54(C-19), 14.63(C-20).

[0131] Yield: 78%, Melting point: 193-194℃, Mass spectrometry data: C 29 H 38 Cl2N4O2[M+H] + Calculated value: 545.2445, measured value: 545.2438.

[0132] Example 11

[0133]

[0134] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3j):

[0135] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-nitrophenyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0136] The NMR data are as follows: 1H NMR (400MHz, CDCl3) δ8.40 (d, J=8.6Hz, 2H, 2×Ph-H), 8.21 (s, 1H, cH), 7.97 (d, J=8.7H z, 2H, 2×Ph-H), 6.39 (s, 1H, NH), 5.56 (s, 1H, 14-H), 4.84 (s, 1H, 17-H), 4.62 (s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.6, 4.3Hz, 1H, 3-H), 2.38 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.30–2.20 (m, 1H, 12-H), 2.15 (s, 3H, 16-C H3 ), 1.99–1.83(m, 2H, 7-H, 12-H), 1.78–1.44(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=12.9, 4.2Hz, 1H, 6-H), 1.14 (td, J=13.0, 3.6Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.66 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.25(C-15), 156.61(C-13), 147.68(C-8), 147.23(Ph-C), 146.63(bC), 141.0 4(Ph-C), 125.50(2×Ph-C), 120.92(cC), 120.49(2×Ph-C), 117.05(C-14), 106.72(C-17), 78.70(C-3), 55.83(C-9), 54.50(C-5), 39.61(C-10), 39.35(C-4), 39.08(C-7), 38.07(C-12), 37.02(C-1), 34.46( aC), 28.26(C-18), 27.82(C-2), 23.94(C-6), 21.68(C-11), 18.50(C-16), 15.39(C-19), 14.46(C-20).

[0137] Yield: 50%, Melting point: 160-162℃, Mass spectrometry data: C 29 H 39 N5O4[M+H]+ Calculated value: 522.3075, measured value: 522.3076.

[0138] Example 12

[0139]

[0140] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3k):

[0141] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-trifluoromethyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0142] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.14 (s, 1H, cH), 7.88 (d, J=8.4Hz, 2H, 2×Ph-H), 7.79 (d, J=8.4H z, 2H, 2×Ph-H), 6.44 (s, 1H, NH), 5.57 (s, 1H, 14-H), 4.84 (s, 1H, 17-H), 4.62 (s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.38 (ddd, J=12.8, 4.3, 2.4Hz, 2H, 7-H), 2.27–2.19 (m, 1H, 12-H), 2.15 (s, 3H, 16-C H3 ), 1.99–1.83(m, 2H, 7-H, 12-H), 1.78–1.46(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.37 (qd, J=12.9, 4.1Hz, 1H, 6-H), 1.14 (td, J=13.2, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.66 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.37(C-15), 156.52(C-13), 147.84(C-8), 146.34(bC), 139.41(P h-C), 130.98 (q, J=32.9Hz, Ph-C), 127.24 (q, J=3.6Hz, 2×Ph-C), 123.62 (q, J=272.3Hz, Ph- C F3), 120.95(cC), 120.58(2×Ph-C), 117.32(C-14), 106.87(C-17), 78.85(C-3), 55.99(C-9), 54.66(C-5), 39.75(C-10), 39.51(C-4),

[0143] 39.24(C-7), 38.23(C-12), 37.17(C-1), 34.55(aC), 28.42(C-18), 27.98(C -2), 24.10(C-6), 21.83(C-11), 18.64(C-16), 15.55(C-19), 14.61(C-20).

[0144] Yield: 92%, Melting point: 193-194℃, Mass spectrometry data: C 30 H 39 F3N4O2[M+H] + Calculated value: 545.3098, measured value: 545.3095.

[0145] Example 13

[0146]

[0147] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphthyl-1-yl)-3-methyl-N-((1-(4-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3l):

[0148] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-methoxyazide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0149] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.98 (s, 1H, cH), 7.63–7.54 (m, 2H, 2×Ph-H), 7.08–6.91 (m, 2 H, 2×Ph-H), 6.64 (s, 1H, NH), 5.58 (s, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.60 (s, 2H, aC H2 ), 4.47(s, 1H, 17-H), 3.85(s, 3H, Ph-OC H3 ), 3.23 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.26–2.17 (m, 1H, 12-H), 2.14 (s, 3H, 16-C H3 ), 1.97–1.80(m, 2H, 7-H, 12-H), 1.77–1.42(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.35 (qd, J=12.9, 12.5, 3.9Hz, 1H, 6-H), 1.12 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ 167.32 (C-15), 160.06 (C-13), 156.08 (Ph-C), 147.80 (C-8), 145.40 (bC), 130.33 (Ph-C), 122 .29(2×Ph-C), 121.22(cC), 117.47(C-14), 114.89(2×Ph-C), 106.85(C-17), 78.78(C-3), 55.94(C-9), 55.74(Ph-O C H3), 54.60(C-5), 39.69(C-10), 39.46(C-4), 39.21(C-7), 38.20(C-12), 37.12(C-1), 34.40(aC), 28.40(C-18), 27.95(C-2), 24.07(C-6), 21.79(C-11), 18.60(C-16), 15.56(C-19), 14.58(C-20).

[0150] Yield: 77%, Melting point: 142-145℃, Mass spectrometry data: C 30 H 42 N4O3[M+H] + Calculated value: 507.3330, measured value: 507.3328.

[0151] Example 14

[0152]

[0153] Preparation of (E)-N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3m):

[0154] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-cyanoazide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally the ethyl acetate was evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0155] The NMR data are as follows: 1H NMR (400MHz, CDCl3) δ8.14 (s, 1H, cH), 7.91 (d, J=8.4Hz, 2H, 2×Ph-H), 7.83 (d, J=8.4H z, 2H, 2×Ph-H), 6.31 (s, 1H, NH), 5.55 (s, 1H, 14-H), 4.84 (s, 1H, 17-H), 4.61 (s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.9Hz, 1H, 7-H), 2.27–2.19 (m, 1H, 12-H), 2.18–2.12 (m, 3H, 16-C H3 ), 1.98–1.84(m, 2H, 7-H, 12-H), 1.79–1.48(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=13.0, 4.2Hz, 1H, 6-H), 1.14 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.17(C-15), 156.46(C-13), 147.68(C-8), 146.55(bC), 139.72(Ph-C), 133.87(2×Ph-C), 125.47(Ph- C N), 120.50(2×Ph-C), 117.63(cC), 117.07(C-14), 112.39(Ph-C), 106.70( C-17), 78.67(C-3), 55.81(C-9), 54.50(C-5), 39.58(C-10), 39.34(C-4), 3 9.08(C-7), 38.07(C-12), 37.00(C-1), 34.46(aC), 28.25(C-18), 27.82(C -2), 23.93(C-6), 21.66(C-11), 18.45(C-16), 15.37(C-19), 14.44(C-20).

[0156] Yield: 83%, Melting point: 161-163℃, Mass spectrometry data: C30 H 39 N5O2[M+H] + Calculated value: 502.3177, measured value: 502.3162.

[0157] Example 15

[0158]

[0159] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methylenedehydronaphth-1-yl)-3-methyl-N-((1-(naphth-1-yl)-1H-1,2,3-triazol-4-yl)methyl)pent-2-enamide (compound 3n):

[0160] 35.7 mg of the intermediate product N-propargyl terbinamide (2) and 0.12 mmol of α-azidonaphthalene were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0161] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.06–7.89 (m, 3H, cH, 2×Ph-H), 7.64–7.45 (m, 5H, 5×Ph-H), 6.5 4(t, J=5.8Hz, 1H, NH), 5.61 (s, 1H, 14-H), 4.82 (s, 1H, 17-H), 4.70 (d, J=5.8Hz, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.22 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.37 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.30–2.18 (m, 1H, 12-H), 2.15 (s, 3H, 16-C H3 ), 1.95–1.84(m, 2H, 7-H, 12-H), 1.78–1.49(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.36 (qd, J=12.9, 4.3Hz, 1H, 6-H), 1.12 (td, J=13.2, 3.9Hz, 1H, 1-H), 1.04 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.97 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.65 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.27(C-15), 155.99(C-13), 147.82(C-8), 145.11(bC), 134.24(Ph-C), 133.71(Ph-C), 130.52(P h-C), 128.50(cC), 128.38(Ph-C), 127.99(Ph-C), 127.17(Ph-C), 125.06(2×Ph-C), 123.62(Ph-C), 122.40(Ph-C), 117.5 3(C-14), 106.84(C-17), 78.77(C-3), 55.96(C-9), 54.62(C-5), 39.68(C-10), 39.47(C-4), 39.21(C-7), 38.22(C-12), 3 7.13(C-1), 34.68(aC), 28.40(C-18), 27.96(C-2), 24.08(C-6), 21.79(C-11), 18.60(C-16), 15.54(C-19), 14.58(C-20).

[0162] Yield: 93%, Mass spectrometry data: C 33 H 42 N4O2[M+H] + Calculated value: 527.3381, measured value: 527.3380.

[0163] Example 16

[0164]

[0165] Preparation of (E)-N-((1-phenyl-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3o):

[0166] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of benzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0167] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.46 (s, 1H, cH), 7.41–7.32 (m, 3H, 3×Ph-H), 7.27–7.24 (m, 2H, 2×Ph-H), 6.13 (t, J=5.8Hz, 1H, NH), 5.51 (s, 1H, 14-H), 5.48 (s, 2H, dC H2 ), 4.84 (s, 1H, 17-H), 4.50 (d, J=5.9Hz, 2H, aC H2 ), 4.49 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.4, 2.5Hz, 1H, 7-H), 2.26–2.15 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 1.98–1.84(m, 2H, 7-H, 12-H), 1.77–1.56(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=13.1, 12.2, 3.5Hz, 1H, 6-H), 1.14 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.5Hz, 1H, 5-H), 0.99 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ167.11(C-15), 155.86(C-13), 147.86(C-8), 145.44(bC), 134.60(Ph-C), 129.26(2 ×Ph-C), 128.94(cC), 128.25(2×Ph-C), 122.18(Ph-C), 117.54(C-14), 106.90(C-17), 78.87(C-3), 55.94 (C-9), 54.66(C-5), 54.36(dC), 39.65(C-10), 39.52(C-4), 39.26(C-7), 38.25(C-12), 37.16(C-1), 34.7 0(aC), 28.42(C-18), 28.01(C-2), 24.12(C-6), 21.79(C-11), 18.55(C-16), 15.55(C-19), 14.63(C-20).

[0168] Yield: 93%, Melting point: 150-151℃, Mass spectrometry data: C 30 H 42 N4O2[M+H] + Calculated value: 491.3381, measured value: 491.3368.

[0169] Example 17

[0170]

[0171] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-methylphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide (compound 3p):

[0172] 35.7 mg of the intermediate product N-propargyl pyridinamide (2) and 0.12 mmol of p-methylbenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0173] The NMR data are as follows: 1H NMR(400MHz,CDCl3)δ7.49(s,1H,c-H),7.16(s,4H,4×Ph-H),6.40(s,1H,NH),5.52(s,1H,14-H),5.44(s,2H,d-C H2 ),4.83(s,1H,17-H),4.50(d,J=3.8Hz,2H,a-C H2 ),4.48(s,1H,17-H),3.24(dd,J=11.7,4.3Hz,1H,3-H),2.38(ddd,J=13.1,4.3,2.4Hz,1H,7-H),2.34(s,3H,Ph-C H3 ),2.25–2.17(m,1H,12-H),2.11(s,3H,16-C H3 ),1.98–1.81(m,2H,7-H,12-H),1.78–1.46(m,7H,1-H,2-C H2 ,6-H,9-H,11-C H2 ),1.37(qd,J=12.8,4.1Hz,1H,6-H),1.13(td,J=13.1,3.7Hz,1H,1-H),1.06(dd,J=12.5,2.7Hz,1H,5-H),0.98(s,3H,18-C H3 ),0.76(s,3H,19-C H3 ),0.66(s,3H,20-C H3 ). 13 C NMR(100MHz,CDCl3)δ167.04(C-15),155.79(C-13),147.68(C-8),144.84(b-C),138.90(Ph-C),131.02(Ph-C),129.81(2×Ph-C),128.22(2×Ph-C),122.37(c-C),117.34(C-14),106.73(C-17),78.68(C-3),55.79(C-9),54.47(C-5),54.29(d-C),39.51(C-10),39.34(C-4),39.09(C-7),38.08(C-12),36.99(C-1),34.20(a-C),28.27(C-18),27.82(C-2),23.94(C-6),21.63(C-11),21.15(Ph- C H3),18.42(C-16),15.42(C-19),14.46(C-20).

[0174] Yield: 77%, Melting point: 162-164℃, Mass spectrometry data: C 31 H 44 N4O2[M+H] + Calculated value: 505.3537, measured value: 505.3535.

[0175] Example 18

[0176]

[0177] Preparation of (E)-N-((1-(4-fluorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3q):

[0178] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-fluorobenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0179] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.56 (s, 1H, cH), 7.31–7.21 (m, 2H, 2×Ph-H), 7.10–7.00 (m, 2H, 2×Ph-H), 6.49 (s, NH), 5.53 (s, 1H, 14-H), 5.46 (s, 2H, dC H2 ), 4.83(s, 1H, 17-H), 4.51(s, 2H, aC H2 ), 4.47 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.38 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.27–2.15 (m, 1H, 12-H), 2.10 (s, 3H, 16-C H3 ), 1.99–1.80(m, 2H, 7-H, 12-H), 1.79–1.45(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.36 (qd, J=12.9, 4.3Hz, 1H, 6-H), 1.13 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18), 0.75 (s, 3H, 19-C H3 ), 0.66 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ 167.24 (C-15), 163.04 (d, J=248.5Hz, Ph-C), 156.11 (C-13), 147.81 (C-8), 145.22 (bC), 130.23 (d, J=8.5Hz, 2×Ph-C), 130.04 (d, J=3.4Hz, Ph-C), 122.71 (cC), 117.38 (C-14), 116.32 (d, J=21.8Hz, 2×Ph-C), 106.86 ( C-17), 78.80(C-3), 55.92(C-9), 54.59(C-5), 53.88(dC), 39.66(C-10), 39.47(C-4), 39.22(C-7), 38.20(C-12), 37.1 2(C-1), 34.27(aC), 28.41(C-18), 27.94(C-2), 24.07(C-6), 21.75(C-11), 18.57(C-16), 15.57(C-19), 14.60(C-20).

[0180] Yield: 58%, Melting point: 144-146℃, Mass spectrometry data: C 30 H 41 FN4O2[M+H] + Calculated value: 509.3287, measured value: 509.3274.

[0181] Example 19

[0182]

[0183] Preparation of (E)-N-((1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3r):

[0184] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-chlorobenzyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0185] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.55 (s, 1H, cH), 7.38–7.28 (m, 2H, 2×Ph-H), 7.23–7.16 (m, 2H, 2×Ph-H), 6.37 (s, 1H, NH), 5.52 (s, 1H, 14-H), 5.45 (s, 2H, dC H2 ), 4.83(s, 1H, 17-H), 4.51(s, 2H, aC H2 ), 4.47 (s, 1H, 17-H), 3.23 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.38 (ddd, J=12.7, 4.3, 2.4Hz, 1H, 7-H), 2.27–2.16 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 1.99–1.81(m, 2H, 7-H, 12-H), 1.77–1.44(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=12.9, 4.3Hz, 1H, 6-H), 1.13 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.75 (s, 3H, 19-C) H3 ), 0.66 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ167.21(C-15), 156.07(C-13), 147.82(C-8), 145.50(bC), 135.08(Ph-C), 132.84( Ph-C), 129.60(4×Ph-C), 129.48(C-), 122.71(cC), 117.41(C-14), 106.86(C-17), 78.81(C-3), 55.94(C -9), 54.62(C-5), 53.77(dC), 39.66(C-10), 39.48(C-4), 39.23(C-7), 38.22(C-12), 37.14(C-1), 34.42 (aC), 28.42(C-18), 27.96(C-2), 24.08(C-6), 21.77(C-11), 18.57(C-16), 15.56(C-19), 14.61(C-20).

[0186] Yield: 57%, Melting point: 106-108℃, Mass spectrometry data: C 30 H 41 ClN4O2[M+H] + Calculated value: 525.2991, measured value: 525.2990.

[0187] Example 20

[0188]

[0189] Preparation of (E)-N-((1-(4-bromophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3s):

[0190] 35.7 mg of the intermediate product N-propargyl pyridinamide (2) and 0.12 mmol of p-bromobenzyl azide were dissolved in a mixture of 2 mL of N,N-dimethylformamide and 2 mL of water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0191] The NMR data are as follows: 1H NMR (400MHz, CDCl3) δ7.53–7.48 (m, 3H, cH, 2×Ph-H), 7.14 (d, J=8.3Hz, 2H, 2×Ph-H), 6.21 (s, 1H, NH), 5.52 (s, 1H, 14-H), 5.44 (s, 2H, dC H2 ), 4.84(s, 1H, 17-H), 4.51(s, 2H, aC H2 ), 4.49 (s, 1H, 17-H), 3.24 (dd, J=11.6, 4.5Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.25–2.19 (m, 1H, 12-H), 2.12 (s, 3H, 16-C H3 ), 1.98–1.86(m, 2H, 7-H, 12-H), 1.77–1.52(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.39 (qd, J=12.9, 4.3Hz, 1H, 6-H), 1.15 (td, J=12.8, 3.8Hz, 2H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.48(C-15), 156.21(C-13), 148.15(C-8), 145.97(bC), 133.90(Ph-C), 132.71(2 ×Ph-C), 130.13(2×Ph-C), 123.40(Ph-C), 122.69(cC), 117.79(C-14), 107.16(C-17), 79.11(C-3), 56.27 (C-9), 54.95(C-5), 53.94(dC), 39.97(C-10), 39.80(C-4), 39.54(C-7), 38.54(C-12), 37.46(C-1), 34.9 1(aC), 28.73(C-18), 28.30(C-2), 24.40(C-6), 22.09(C-11), 18.87(C-16), 15.86(C-19), 14.92(C-20).

[0192] Yield: 66%, Melting point: 163-164℃, Mass spectrometry data: C 30H 41 BrN4O2[M+H] + Calculated value: 571.2466, measured value: 571.2455.

[0193] Example 21

[0194]

[0195] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide (compound 3t):

[0196] 35.7 mg of the intermediate product N-propargyl terbinamide (2) and 0.12 mmol of p-nitrobenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0197] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ8.24 (d, J=8.2Hz, 2H, 2×Ph-H), 7.70 (s, 1H, cH), 7.43 (d, J=8.3Hz, 2H, 2×Ph-H), 6.39 (s, 1H, NH), 5.63 (s, 2H, dC H2 ), 5.53(s, 1H, 14-H), 4.84(s, 1H, 17-H), 4.56(s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.6, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.29–2.15 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 1.98–1.82(m, 2H, 7-H, 12-H), 1.79–1.48(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2), 1.38 (ddd, J=25.4, 12.6, 3.9Hz, 1H, 6-H), 1.14 (td, J=13.2, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.99 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13 CNMR (100MHz, CDCl3) δ167.01(C-15), 156.26(C-13), 148.02(Ph-C), 147.51(C-8), 145.28(bC), 140.73( Ph-C), 128.64(2×Ph-C), 124.22(2×Ph-C), 123.09(cC), 116.94(C-14), 106.58(C-17), 78.56(C-3), 55.6 5(C-9), 54.33(C-5), 53.39(dC), 39.40(C-10), 39.20(C-4), 38.94(C-7), 37.92(C-12), 36.86(C-1), 34. 01(aC), 28.11(C-18), 27.67(C-2), 23.78(C-6), 21.49(C-11), 18.30(C-16), 15.24(C-19), 14.31(C-20).

[0198] Yield: 51%, Melting point: 132-134℃, Mass spectrometry data: C 30 H 41 N5O4[M+H] + Calculated value: 535.3232, measured value: 535.3226.

[0199] Example 22

[0200]

[0201] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphth-1-yl)-3-methyl-N-((1-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide (compound 3u):

[0202] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-trifluoromethylbenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0203] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.64 (s, 1H, cH), 7.62 (d, J=4.8Hz, 2H, 2×Ph-H), 7.38 (d, J=8.0Hz, 2H, 2×Ph-H), 6.40 (s, 1H, NH), 5.56 (s, 2H, dC H2 ), 5.53(s, 1H, 14-H), 4.84(s, 1H, 17-H), 4.53(s, 2H, aC H2 ), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.2, 2.4Hz, 1H, 7-H), 2.28–2.16 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 2.00–1.80(m, 2H, 7-H, 12-H), 1.80–1.46(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=12.8, 4.1Hz, 1H, 6-H), 1.14 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CDCl3) δ167.32(C-15), 156.31(C-13), 147.84(C-8), 145.43(bC), 138.07(Ph-C), 1 31.40 (q, J=32.4Hz, Ph-C), 127.89 (cC), 126.33 (q, J=3.7Hz, 2×Ph-C), 123.83 (q, J=272.2Hz, Ph- C F3), 123.09(2×Ph-C), 117.36(C-14), 106.89(C-17), 78.88(C-3), 55. 99(C-9), 54.66(C-5), 54.01(dC), 39.71(C-10), 39.51(C-4), 39.25(C -7), 38.24(C-12), 37.17(C-1), 34.26(aC), 28.42(C-18), 27.99(C-2) , 24.11(C-6), 21.82(C-11), 18.59(C-16), 15.56(C-19), 14.62(C-20).

[0204] Yield: 81%, Melting point: 146-147℃, Mass spectrometry data: C 31 H 41 F3N4O2[M+H] + Calculated value: 559.3254, measured value: 559.3235.

[0205] Example 23

[0206]

[0207] Preparation of (E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methyl-N-((1-(4-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentan-2-amide (compound 3v):

[0208] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-methoxybenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0209] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.49 (s, 1H, cH), 7.24–7.20 (m, 2H, 2×Ph-H), 6.91–6.87 (m, 2H, 2×Ph-H), 6.34 (s, 1H, NH), 5.52 (s, 1H, 14-H), 5.43 (s, 2H, dC H2 ), 4.84 (s, 1H, 17-H), 4.51 (d, J=5.1Hz, 2H, aC H2 ), 4.48(s, 1H, 17-H), 3.80(s, 3H, Ph-OC H3 ), 3.24 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.27–2.16 (m, 1H, 12-H), 2.12–2.09 (m, 3H, 16-C H3 ), 1.99–1.83(m, 2H, 7-H, 12-H), 1.79–1.51(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=12.9, 4.3Hz, 1H, 6-H), 1.14 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.67 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.18(C-15), 160.24(C-13), 156.15(Ph-C), 147.82(C-8), 144.78(bC), 130.00(2×Ph-C), 125.88 (Ph-C), 122.55(cC), 117.41(C-14), 114.71(2×Ph-C), 106.90(C-17), 78.85(C-3), 55.92(C-9), 55.50(C-5), 54.61(Ph-O CH3), 54.41(dC), 39.67(C-10), 39.49(C-4), 39.24(C-7), 38.22(C-12), 37.13(C-1), 34.17(aC), 28.42(C-18), 27.98(C-2), 24.09(C-6), 21.77(C-11), 18.59(C-16), 15.56(C-19), 14.62(C-20).

[0210] Yield: 55%, Melting point: 116-117℃, Mass spectrometry data: C 31 H 44 N4O3[M+H] + Calculated value: 521.3287, measured value: 521.3272.

[0211] Example 24

[0212]

[0213] Preparation of (E)-N-((1-(4-cyanophenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpent-2-enamide (compound 3w):

[0214] 35.7 mg of the intermediate product N-propargyl ferrugin (2) and 0.12 mmol of p-cyanobenzyl azide were dissolved in a mixture of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0215] The NMR data are as follows: 1 H NMR (400MHz, CDCl3) δ7.66 (d, J=7.9Hz, 2H, 2×Ph-H), 7.63 (s, 1H, cH), 7.34 (d, J=8.0Hz, 2H, 2×Ph-H), 6.39 (s, 1H, NH), 5.56 (s, 2H, dC H2 ), 5.53(s, 1H, 14-H), 4.84(s, 1H, 17-H), 4.52(s, 2H, aC H2), 4.48 (s, 1H, 17-H), 3.24 (dd, J=11.6, 4.3Hz, 1H, 3-H), 2.38 (ddd, J=12.7, 4.2, 2.4Hz, 1H, 7-H), 2.28–2.16 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 1.97–1.81(m, 2H, 7-H, 12-H), 1.78–1.46(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.37 (qd, J=12.8, 4.2Hz, 1H, 6-H), 1.13 (td, J=13.3, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-C H3 ), 0.76 (s, 3H, 19-C) H3 ), 0.66 (s, 3H, 20-C) H3 ). 13 C NMR (100MHz, CDCl3) δ167.28(C-15), 156.30(C-13), 147.83(C-8), 145.72(bC ), 139.46(Ph-C), 133.04(2×Ph-C), 128.65(2×Ph-C), 123.13(cC), 118.17(Ph- C N), 117.32(C-14), 113.06(Ph-C), 106.86(C-17), 78.83(C-3), 55.97(C-9), 54.65(C-5), 53.77(dC), 39.69(C-10), 39.50(C-4), 39.24(C-7 ), 38.22(C-12), 37.16(C-1), 34.41(aC), 28.42(C-18), 27.97(C-2), 24.09(C-6), 21.80(C-11), 18.59(C-16), 15.56(C-19), 14.61(C-20).

[0216] Yield: 67%, Melting point: 125-127℃, Mass spectrometry data: C 31 H 41 N5O2[M+H] + Calculated value: 516.3334, measured value: 516.3322.

[0217] Example 25

[0218]

[0219] Preparation of (4-((4-(((E)-5-([1R,4aS,6R,8aS]-6-hydroxy-5,5,8a-trimethyl-2-methyldecahydronaphthyl-1-yl)-3-methylpentan-2-amido)methyl)-1H-1,2,3-triazol-1-yl)methyl)phenyl)boronic acid (compound 3x):

[0220] 35.7 mg of the intermediate product N-propargyl terbinamide (2) and 0.12 mmol of p-azidomethylphenylboronic acid were dissolved in a mixed solution of 2 mL N,N-dimethylformamide and 2 mL water. Then, 7.8 mg of sodium ascorbate and 4.5 mg of anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60 °C until the reaction was complete. The reaction was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and finally evaporated to dryness. The crude product obtained after evaporation was purified by silica gel column chromatography to obtain the target compound.

[0221] The NMR data are as follows: 1 H NMR (400MHz, CD3OD) δ7.85 (s, 1H, cH), 7.81–7.25 (m, 4H, 4×Ph-H), 5.68 (s, 1H, 14-H), 5.59 (s, 2H, dC H2 ), 4.88(s, 1H, 17-H), 4.56(s, 1H, 17-H), 4.46(s, 2H, aC H2 ), 3.26–3.16 (m, 1H, 3-H), 2.43 (ddd, J=12.7, 4.3, 2.4Hz, 1H, 7-H), 2.31–2.20 (m, 1H, 12-H), 2.11 (s, 3H, 16-C H3 ), 2.02–1.91(m, 2H, 7-H, 12-H), 1.86–1.53(m, 7H, 1-H, 2-C H2 6-H, 9-H, 11-C H2 ), 1.42 (qd, J=12.9, 4.2Hz, 1H, 6-H), 1.21 (td, J=8.6, 7.2, 4.9Hz, 1H, 1-H), 1.13 (dd, J=12.6, 2.7Hz, 1H, 5-H), 1.00 (s, 3H, 18-C H3 ), 0.79 (s, 3H, 19-C) H3 ), 0.74 (s, 3H, 20-C) H3 ). 13C NMR (100MHz, CD3OD) δ169.59(C-15), 156.41(C-13), 149.31(C-8), 146.86(bC), 135.15(Ph-C), 130.84(c C), 128.24(Ph-C), 124.25(2×Ph-C), 118.70(2×Ph-C), 116.65(C-14), 107.19(C-17), 79.41(C-3), 57.07 (C-9), 55.97(C-5), 54.86(dC), 40.53(C-10), 40.40(C-4), 40.18(C-7), 39.27(C-12), 38.30(C-1), 35.3 2(aC), 28.87(C-18), 28.60(C-2), 25.23(C-6), 22.83(C-11), 18.64(C-16), 16.13(C-19), 15.03(C-20).

[0222] Yield: 43%, Melting point: 260-262℃, Mass spectrometry data: C 30 H 43 BN4O4[M+H] + Calculated value: 535.3451, measured value: 535.3441.

[0223] The antitumor activity of 24 synthesized derivatives 3a-3x was studied.

[0224] 2. Experimental Procedures for Antitumor Activity

[0225] (1) Observe the cell state under an optical microscope, select a cell line in the logarithmic growth phase, and follow the same cell passage method. First, wash the cells once with sterile PBS, then digest them with trypsin for a few minutes (depending on the time it takes for the cells to become rounded after digestion). Centrifuge to collect the cells, resuspend them in 1 mL of DMEM complete medium, and thoroughly pipette to prepare a single-cell suspension. Add 100 μL of cell suspension to each well of a 96-well plate, count the cells, and then seed the plate to adjust the cell density to 1000-10000 cells / well (fill the edge wells with sterile PBS). Incubate the 96-well plate in a 37°C, 5% CO2 cell culture incubator.

[0226] (2) After culturing for about 24 hours, the cells are fully attached to the wall. Determine the drug concentration gradient, which is generally 5-6 wells. Set up 3 replicates for each group and test three groups of single drugs.

[0227] (3) Incubate at 37°C for 72 hours with 5% CO2 and observe under an inverted microscope.

[0228] (4) Add 20 μL of 5 mg / mL MTT solution to each well and incubate at 37°C in a 5% CO2 cell culture incubator for 2 h.

[0229] (5) After 2 hours, succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple formazan crystals, which are then deposited in the cells. Dead cells do not have this function. Therefore, carefully discard the original culture medium in each well, add 100 μL of triple solution to each well, and incubate overnight in a 5% CO2, 37°C incubator to allow the crystals to dissolve completely. Measure the absorbance of each well at 570 nm using an ELISA reader.

[0230] (6) Simultaneously set up zeroing wells (culture medium, MTT, dimethyl sulfoxide) and control wells (cells, drug dissolution medium of the same concentration, culture medium, MTT, dimethyl sulfoxide).

[0231] The IC50 values ​​of each compound against human non-small cell lung cancer cells A549, human lung cancer cells PC-9, human breast cancer cells MCF-7, human prostate cancer cells PC-3, human cervical cancer cells HeLa, and human normal liver cells HL-7702 were [not specified]. 50 As shown in Table 1.

[0232] Table 1. Results of antitumor activity of compounds (3a-3x)

[0233]

[0234]

[0235] This invention uses fern fiber as a substrate, first condensing it with propyneamine to obtain the intermediate N-propylated fern fiber amide, then reacting it with an azide compound under copper(I) ion catalysis to obtain a series of 1,2,3-triazole derivatives of fern fiber. These compounds were structurally identified by physicochemical properties and various spectroscopic methods. In the antitumor activity screening, cisplatin was used as a positive control, and its cytotoxicity against human non-small cell lung cancer cells A549, human lung cancer cells PC-9, human breast cancer cells MCF-7, human prostate cancer cells PC-3, human cervical cancer cells HeLa, and human normal liver cell line HL-7702 was determined using the MTT assay.

[0236] In the screening for antitumor activity, most compounds showed significant antitumor activity, with some compounds exhibiting particularly pronounced antitumor effects, such as compounds 3b, 3n, 3o, 3q, 3r, 3s, and 3u. For most cancer cells in these five different human cancer cell lines, the IC50 concentration was [missing information]. 50 The values ​​were all around 10 μM. Among these, compound 3n showed an IC50 value for A549 and PC-9 cells. 50The concentrations were 4.0 μM and 7.23 μM, respectively; the IC50 values ​​of compound 3s for MCF-7 cells, PC-3 cells, and HeLa cells were [not specified]. 50 The values ​​were 2.12 μM, 5.74 μM, and 7.58 μM, respectively. Compared with the other five compounds, these two compounds had relatively low cytotoxicity to normal human hepatocytes (HL-7702) and high selectivity between normal human hepatocytes and cancer cells. Therefore, 3n and 3s are the two compounds with better anti-tumor effects.

[0237] In the description of this specification, the references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0238] The above description is merely an example and illustration of the present invention. Those skilled in the art can make various modifications or additions to the specific embodiments described, or use similar methods to replace them, as long as they do not deviate from the invention or exceed the scope defined in the claims, all of which should fall within the protection scope of the present invention.

Claims

1. A fern acid derivative containing a 1,2,3-triazole group, characterized in that, The structure of the derivative is shown below: 。 2. The method for preparing the fern acid derivative according to claim 1, characterized in that, The specific steps are as follows: (1) Dissolve the powdered fern fiber in dichloromethane, then add diisopropylethylamine in sequence. N , N , N' , N' -Tetramethylurea hexafluorophosphate and propargylamine were stirred at room temperature until the reaction was complete. The reaction was then quenched with water, extracted, washed, and the organic phase was dried under reduced pressure to obtain the crude product. The crude product was then separated and purified by silica gel column chromatography to obtain the intermediate product. N -Propylpyridinium trifoliate; (2) The result obtained in step (1) N - Acrylaryl fernamide and organic azide added N , N The product was dissolved in a mixed solution of dimethylformamide and water, and then sodium ascorbate and anhydrous copper sulfate were added sequentially. The reaction system was stirred at 60°C. After the reaction was complete, the product was extracted, washed, dried, and evaporated to dryness to obtain the crude product. The crude product was then separated and purified by silica gel column chromatography to obtain the target compound.

3. The application of the fern acid derivative according to claim 1 in the manufacture of anti-prostate cancer drugs.

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