Dehydroabietic acid 1, 2, 3-triazole derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of organic chemistry, and particularly relates to a dehydroabietic acid 1, 2, 3-triazole derivative as well as a preparation method and application thereof. The preparation method comprises the following steps: reacting dehydroabietic acid in a first organic solvent under the action of a reducing agent to obtain a compound 2 with reduced carboxyl; reacting the compound 2 with p-toluenesulfonyl chloride under the action of an alkali solution to obtain a hydroxyl-protected compound 3; in a second organic solvent, reacting the compound 3 with sodium azide under a high-temperature condition to obtain an azide compound 4; and reacting the compound 4 with a copper salt and an alkyne compound in a polar solvent of alkali to obtain a target compound. The dehydroabietic acid derivative with high biological activity is synthesized, raw materials are cheap and easy to obtain, reaction conditions are mild, aftertreatment is simple, the yield is high, a virulent solvent is not used in the synthesis process, the green chemical requirement is met, high pressure or a special catalyst is not needed, and large-scale production can be achieved in conventional equipment.

Description

Technical Field

[0001] This invention belongs to the field of organic chemistry technology, and specifically relates to a dehydrorosinic acid 1,2,3-triazole derivative, its preparation method and application. Background Technology

[0002] Cancer, also known as malignant tumor, is a disease caused by the abnormal proliferation of cells. It is invasive and metastatic, and as a global health challenge, its incidence, treatment methods, and social impact are complex and ever-changing. In recent years, the incidence of cancer has continued to rise, becoming one of the leading causes of death worldwide. According to data from the World Health Organization (WHO), cancer is the second leading cause of death globally, second only to cardiovascular disease. With the advancement of medical technology, cancer treatment methods are constantly being updated and improved, including surgery, radiotherapy, chemotherapy, immunotherapy, and drug therapy, among which drug therapy remains the most important method for treating cancer. Therefore, the development of highly effective and low-toxicity anticancer drugs has become a key focus of research, especially the search for low-toxicity and highly effective anticancer active ingredients from natural animals and plants, as well as further structural modifications of the effective components of natural drugs to semi-synthesize derivatives with better anticancer activity. These have become hot topics for researchers both domestically and internationally.

[0003] Natural products, with their extensive structural diversity and pharmacological multifunctionality, are an important source for discovering lead compounds and have a profound impact on drug development. Dehydroabietic acid (DHAA), also known as dehydroabsionic acid, has the chemical structure shown below. It is generally derived from abietic resin acids in rosin through catalytic dehydrogenation. Compared to abietic acids, it has an additional conjugated double bond in its molecule, significantly improving its thermal stability, antioxidant properties, and chemical activity, while retaining the viscosity and film-forming properties of rosin. It is an important natural triterpenoid resin acid with broad pharmacological activities, including antibacterial, antifungal, antiviral, and anti-inflammatory activities. Particularly in terms of antitumor activity, it is a lead compound with potential antitumor effects.

[0004]

[0005] Chemical structural formula of dehydrorosin acid

[0006] In recent years, dehydrorosinic acid derivatives obtained through appropriate structural modifications have exhibited various antitumor activities, including against lung cancer, liver cancer, breast cancer, glioma, and ovarian cancer. Huang reported a series of thiourea α-aminophosphonate derivatives containing dehydrorosinic acid and evaluated the inhibitory activity of the targeted compounds against various human cancer cell lines (human lung cancer cells NCI-H460 and A549, human liver cancer cells HepG2, and human ovarian cancer cells SKOV3) using the MTT assay. The antitumor activity of most of the targeted compounds was higher than that of the commercial anticancer drug 5-fluorouracil. Chen designed and synthesized a series of novel dehydrorosinic acid 1-H-dibenzo[a,c]carbazole derivatives with different N-(piperazin-1-yl)alkyl side chains, which showed the strongest proliferation inhibitory activity against three types of human liver cancer cells (SMMC-7721, HepG2, and Hep3B). Kovaleva et al. synthesized a series of rosin diterpenoid ureas, thioureas, amides, and thioamide derivatives containing adamantyl groups and evaluated their inhibitory activity against tyrosine-DNA phosphodiesterase I (TDP-I). The synthesized compounds showed inhibitory activity against TDP-I in the micromolar concentration range (0.19–2.3 μM) and good anti-proliferative activity against T98G glioma cells.

[0007] Currently, the variety of dehydrorosin acid derivatives is limited, and there is little room for performance improvement. Most research on dehydrorosin acid relies on traditional reactions such as esterification and amidation, lacking innovative synthetic methods (e.g., click chemistry, green catalysis). Furthermore, dehydrorosin acid itself has limited reaction sites; the most easily modified sites are the two double bonds at positions 7 and 14, and the carboxyl group at position 18, making it difficult to modify and obtain novel derivatives. The preparation process involves high temperatures, harsh reaction conditions, and often uses highly toxic solvents (such as benzene). In addition, research on dehydrorosin acid and its derivatives is insufficient in terms of basic mechanisms, application development, and technological industrialization. Although dehydrorosin acid has great potential for chemical modification, existing derivatives lack biological activity (e.g., antibacterial, antitumor), or have low activity, limiting its rational design for pharmaceutical or high-performance materials. Therefore, developing novel dehydrorosin acid derivatives to achieve high-value applications while aligning with the trends of green chemistry and sustainable development is of great significance. Summary of the Invention

[0008] To address the shortcomings of existing methods, this invention provides a dehydrorosinic acid 1,2,3-triazole derivative, its preparation method, and its application.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] A dehydrorosinic acid 1,2,3-triazole derivative has the following chemical structural formula:

[0011]

[0012] In the formula, R represents an alkyl or substituted phenyl group.

[0013] Preferably, the alkyl group is n-pentyl, 3-bromopropyl, n-butyl, 5-hydroxypentyl, 1-methylbutyl, 4-chlorobutyl, or 3-cyanopropyl;

[0014] The substituted phenyl group is 4-methoxyphenyl, 2-fluorophenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2-bromophenyl, 3-bromophenyl, phenyl, 4-trifluoromethylphenyl, 4-nitrophenyl, or 4-methylphenyl.

[0015] A method for preparing a dehydrorosinic acid 1,2,3-triazole derivative includes the following steps:

[0016] (1) In the first organic solvent, dehydrorosin acid reacts under the action of a reducing agent. After the reaction is completed, an alkaline aqueous solution is added to quench the reaction. Then, the mixture is extracted and separated to obtain compound 2 with reduced carboxyl groups.

[0017] (2) Compound 2 reacts with p-methylbenzenesulfonyl chloride in an alkaline solution. After the reaction is completed, an acidic buffer solution is added to quench the reaction. Then, the mixture is extracted and separated to obtain compound 3 with the hydroxyl group protected.

[0018] (3) In the second organic solvent, compound 3 reacts with sodium azide under high temperature conditions. After the reaction is completed, an acidic aqueous solution is added to quench the reaction, and then the mixture is extracted and separated to obtain azide compound 4.

[0019] (4) Compound 4 and copper salt react with alkyne compound in a polar alkaline solvent. After the reaction is completed, water is added to quench the reaction, followed by extraction and separation to obtain the target compound.

[0020] Preferably, the molar ratio of dehydrorosin acid and reducing agent in step (1) is 1:2-6.

[0021] Preferably, in step (1), the first organic solvent is one or more of diethyl ether, tetrahydrofuran, 1,4-dioxane, methyl tert-butyl ether, and acetonitrile;

[0022] The reducing agent is one or more of lithium aluminum hydride, sodium borohydride and sodium cyanoborohydride;

[0023] The alkaline aqueous solution is a sodium hydroxide aqueous solution with a mass concentration of 10-20%, a sodium bicarbonate aqueous solution with a mass concentration of 20-40%, or a sodium carbonate aqueous solution with a mass concentration of 15-25%.

[0024] Preferably, the molar ratio of compound 2 and p-methylbenzenesulfonyl chloride in step (2) is 1:2-7.

[0025] Preferably, the alkaline solution in step (2) is one or more of triethylamine, triethanolamine, and pyridine;

[0026] The acidic buffer solution is one or more of the following: saturated ammonium chloride solution, acetic acid-sodium acetate solution, and citric acid-sodium citrate solution.

[0027] Preferably, the molar ratio of compound 3 to sodium azide in step (3) is 1:2-6; the temperature of the high-temperature condition is 40℃-100℃.

[0028] Preferably, in step (3), the second organic solvent is one or more of hexamethylphosphoric triamine, N,N-dimethylformamide and dimethyl sulfoxide;

[0029] The acidic aqueous solution is a 5-15% sulfuric acid aqueous solution, a 5-15% hydrochloric acid aqueous solution, or a 5-15% nitric acid aqueous solution.

[0030] Preferably, the molar ratio of compound 4, copper salt, and alkyne compound in step (4) is 1:0.2-1:1-3.

[0031] Preferably, the copper salt in step (4) is one or more of copper sulfate, copper acetate, copper chloride, copper nitrate and copper carbonate;

[0032] The alkali is one or more of sodium carbonate, sodium bicarbonate, sodium sulfite, sodium bisulfite, and sodium ascorbate;

[0033] The polar solvent is one or more of water-methanol, water-ethanol, water-isopropanol, water-n-butanol, and water-tert-butanol;

[0034] The alkyne compound is one or more of the following: heptynylene, 5-bromopentyne, 1-hexyne, 6-heptyne alcohol, 3-methyl-1-hexyne, 6-chlorohexyne, hexynenitrile, p-methoxyphenylacetylene, 2-fluorophenylacetylene, 4-fluorophenylacetylene, 3-chlorophenylacetylene, 4-chlorophenylacetylene, 2-bromophenylacetylene, 3-bromophenylacetylene, phenylacetylene, 4-trifluoromethylphenylacetylene, 4-nitrophenylacetylene, and 4-methylphenylacetylene.

[0035] Application of a dehydrorosinic acid 1,2,3-triazole derivative in the preparation of drugs for treating cancer.

[0036] Preferably, the cancer is human cervical cancer cells (HeLa), human breast cancer cells (MCF-7), or human gastric cancer cells (HGC-27).

[0037] The reaction equation for the preparation method of the dehydrorosinic acid 1,2,3-triazole derivative of this invention is as follows:

[0038]

[0039] The definition of R is the same as that in the chemical structural formula of the dehydrorosinic acid 1,2,3-triazole derivative.

[0040] The positive and beneficial effects of this invention are as follows:

[0041] 1. This invention uses dehydrorosinic acid as the parent compound and modifies it with 1,2,3-triazole fragments with different substituents. Through reduction, esterification, azidation, and triazole click reactions, the 18-carboxyl group of dehydrorosinic acid is modified, synthesizing a dehydrorosinic acid derivative with high biological activity. The antitumor effect is significantly better than that of dehydrorosinic acid and cisplatin, providing a research basis and lead compound for new drug screening, and has good application prospects.

[0042] 2. This invention also discloses a method for preparing dehydrorosinic acid 1,2,3-triazole derivatives. The raw materials are inexpensive and readily available, the reaction conditions are mild, the post-processing is simple, the yield is high, and no highly toxic solvents (such as benzene) are used in the synthesis process. The solvents can be recycled, which meets the requirements of green chemistry. Furthermore, no high pressure or special catalysts are required, and large-scale production can be achieved in conventional equipment, which meets the needs of industrial development.

[0043] 3. This invention introduces a 1,2,3-triazole heterocyclic structural unit with strong anticancer activity into the molecular backbone of dehydrorosinic acid. By testing the inhibitory activity of dehydrorosinic acid derivatives on human cervical cancer cells (HeLa), human breast cancer cells (MCF-7), and human renal epithelial cells (HEK-293T), highly active lead compounds are screened out, providing a research basis and lead compounds for the development of new anticancer drugs, and showing good application prospects. Attached Figure Description

[0044] Figure 1 This is the hydrogen spectrum of compound 2 of the present invention;

[0045] Figure 2 This is the carbon spectrum of compound 2 of the present invention;

[0046] Figure 3 This is the hydrogen spectrum of compound 3 of the present invention;

[0047] Figure 4 This is the carbon spectrum of compound 3 of the present invention;

[0048] Figure 5 This is the hydrogen spectrum of compound 4 of the present invention;

[0049] Figure 6 This is the carbon spectrum of compound 4 of the present invention;

[0050] Figure 7 This is the 5c proton NMR spectrum of the compound of this invention;

[0051] Figure 8 This is the 5c carbon spectrum of the compound of this invention;

[0052] Figure 9 This is the 5d proton NMR spectrum of the compound of this invention;

[0053] Figure 10 This is the 5d carbon spectrum of the compound of this invention;

[0054] Figure 11 This is the proton NMR spectrum of compound 5j of the present invention;

[0055] Figure 12 This is the carbon spectrum of compound 5j of the present invention. Detailed Implementation

[0056] The present invention will be further described below with reference to some specific embodiments.

[0057] Example 1: Preparation of Compound 2

[0058]

[0059] Compound 1, dehydrorosinic acid (5.0 g, 16.65 mmol), was dissolved in 30 mL of dry THF. Then, a THF solution of LiAlH4 (3.2 g, 83.5 mmol) (20 mL) was added dropwise at 0 °C. The reaction was allowed to proceed for 0.5 h, followed by 2 h at room temperature until the reaction was complete. Thin-layer chromatography was used for detection. After the reaction was complete, a 30% sodium bicarbonate aqueous solution (100 mL) was slowly added to the reaction mixture at 0 °C for quenching. Then, ethyl acetate (3 × 100 mL) was added for extraction. The combined organic phases were washed successively with saturated NaHCO3 aqueous solution (100 mL) and saturated brine (50 mL), dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by column chromatography (SiO2, ethyl acetate:heptane = 1:3) to obtain compound 2.

[0060] Compound 2: The proton and carbon spectra are shown below. Figure 1 and 2 White solid, yield 91.0%, mp 85-87℃; [α] 20 D = -77.4 (c 1.0, CHCl3); 1 H NMR (400MHz, CDCl3) δ7.11 (d, J = 8.2Hz, 1H, H 14 ), 6.92 (d, J = 8.1 Hz, 1H, H 11 ),6.81(s,1H,H 12 ), 3.40 (d, J = 10.9 Hz, 1H, H 18), 3.16(d, J = 10.9 Hz, 1H, H 18 ),2.90–2.64(m,3H,H7,H 15 ),2.27–1.96(m,2H,H1),1.16(dd,J=10.8,7.9Hz,17H),0.82(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ147.31,145.52,134.74,126.79,124.22,123.78,77.32,77.00,76.68,72.26 ,43.96,38.45,37.84,35.11,33.44,30.09,25.25,23.98,23.96,18.86,18.64,17.38,1.01; FT-IRν max ,cm -1 :3568.54(OH),3212.32,3021.77(Ph-H),1619.26(Ph),1564.89,1360.26,1025.45,986.54,835.45,821.54; HR-MS(ESI-TOF)m / z[M+H]+;C 20 H 30 O;Calculated:287.2364;Found:287.2369.

[0061] Example 2: Preparation of Compound 3

[0062]

[0063] In an ice-water bath, p-toluenesulfonyl chloride (13.0 g, 68.0 mmol) was added to 100 mL of a pyridine solution of compound 2 (3.24 g, 11.3 mmol). The mixture was stirred at 0 °C for 0.5 h and then stirred overnight at room temperature. Thin-layer chromatography was used for detection. After the reaction was complete, 100 mL of ice-saturated ammonium chloride solution was added to the mixture for quenching. The mixture was then extracted with ethyl acetate (3 × 50 mL). The combined organic phases were washed successively with hydrochloric acid aqueous solution (1 mol / L, 3 × 50 mL), water (3 × 50 mL), and saturated brine (3 × 50 mL). The mixture was dried over anhydrous Na₂SO₄, concentrated under reduced pressure, and purified by column chromatography (SiO₂, ethyl acetate:heptane = 1:20) to give a white solid compound 3.

[0064] Compound 3: The proton and carbon spectra are shown below. Figure 3 and 4 White solid, yield 94.0%. [α] 20 D= -24.9 (c1.0, CHCl3); 1 H NMR(400MHz,CDCl3)δ7.69(s,1H,H Ph ),7.66(s,1H,H Ph ), 7.23(d, J = 8.1 Hz, 2H, H Ph ), 7.05 (d, J = 8.2 Hz, 1H, H 14 ), 6.88 (d, J = 8.1 Hz, 1H, H) 11 ),6.77(s,1H,H 12 ), 3.73 (d, J = 9.3 Hz, 1H, H 18 ), 3.50(d, J = 9.3 Hz, 1H, H 18 ),2.77–2.60(m,3H,H7,H 15 ),2.34(s,3H,H Ph-CH3 ),1.62–1.50(m,4H),1.34–1.21(m,4H),1.15–1.05(m,10H),0.78(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ146.65,145.50,144.55,134.34,132.88,129.70,127.77,126.65,124.07,123.76,77.63,77.32, 77.00,76.68,43.49,37.93,37.21,37.03,34.94,33.33,29.72,25.08,23.88,21.51,18.72,18.22,17.00,0.92; FT-IRν max ,cm -1 :3601.32,3045.25(Ph-H),2603.21,2354.23,1815.02,1576.89(Ph),1475.6 5,1326.32,1074.65,996.05,854.32,687.65; HR-MS(ESI-TOF)m / z[M+Na]+; C 27 H 36 O3SNa;Calculated:463.2272;Found:463.2277.

[0065] Example 3: Preparation of Compound 4

[0066]

[0067] Compound 3 (3.0 g, 6.82 mmol) was added to 100 mL of dimethyl sulfoxide solution of NaN3 (20.22 g, 34.1 mmol) at room temperature. The mixture was reacted at 60 °C for 48 hours and detected by thin-plate chromatography. After the reaction was completed, the mixture was cooled to room temperature and quenched with 10% aqueous H2SO4 solution (100 mL). The mixture was then extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with saturated bicarbonate solution (2 × 50 mL) and saturated brine (2 × 50 mL), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (SiO2, petroleum ether) to give compound 4 as a white solid.

[0068] Compound 4: The proton and carbon spectra are shown below. Figure 5 and 6 White solid, yield 81.2%, mp 80-81℃; [α] 20 D = -62.5 (c 1.0, CHCl3); 1 H NMR(400MHz, CDCl3)δ7.11–7.04(m,1H,H 14 ), 6.90 (d, J = 8.1 Hz, 1H, H 11 ), 6.80(s, 1H, H 12 ),3.19(d,J=12.1Hz,1H),2.91(d,J=12.1Hz,1H),2.85–2.68(m,3H,H7,H 15 ),2.19(d,J=12.8Hz,1H),1.70–1.52(m,5H),1.33(qd,J=12.7,3.8Hz,3H),1.14(dd,J=15.7,8.8Hz,9H),0.84(s,3H,H 19 ); 13 C NMR (101MHz, CDCl3) δ146.92,145.54,134.46,126.77,124.19,123.85,77.32,77.00,76.68,63.21, 44.78,38.26,38.10,37.35,36.03,33.41,29.96,25.13,23.94,23.93,18.87,18.59,18.37.FT-IRν max ,cm -1:3296.64,3054.24(Ph-H),2506.03,2489.78(azide),2250.37,1608.07( Ph), 1636.17, 1487.08 (N=N), 1396.65, 1006.22, 910.19, 847.30, 7654.43.

[0069] Example 4: Preparation of compounds 5a-5l

[0070]

[0071] CuSO4 (33.4 mg, 0.17 mmol), sodium ascorbate (63.3 mg, 0.32 mmol), and H2O (5 mL) were added to a 50 mL sealed tube and stirred until dissolved. Then, azide compound 4 (0.24 g, 0.77 mmol), t-BuOH (10.0 mL), and alkyne compound (0.84 mmol) were added sequentially. The mixture was stirred overnight at room temperature under sealed conditions. Thin-layer chromatography was used for detection. After the reaction was completed, the reactants were quenched with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with saturated brine (2 × 10 mL), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain white solid compound 5a-5l.

[0072] Compound 5a: colorless oily liquid, yield 85.20%; [α] 20 D = -28.7 (c 1.0, CHCl3); 1 HNMR(400MHz,CDCl3)δ7.11(s,1H,H 21 ), 7.04 (d, J = 8.2 Hz, 1H, H 14 ), 6.88(dd, J = 8.2, 1.5 Hz, 1H, H 11 ), 6.80(s, 1H, H 12 ), 4.21(d, J = 14.0 Hz, 1H, H 18 ), 3.97 (d, J = 14.0 Hz, 1H, H 18 ),2.90–2.78(m,2H,H7),2.76–2.70(m,1H,H 15 ),1.33–1.08(m,31H),0.97(s,3H,H 19 ); 13C NMR (100MHz, CDCl3) δ147.57,146.61,145.55,134.41,126.83,124.03,123.76,122.27,77.32,77.00,76.68,60.75,44.99,38.07 ,37.93,37.48,36.40,33.32,31.35,29.88,28.99,28.92,25.49,23.87,23.84,22.30,19.16,18.64,18.42,13.94,13.90; FT-IRν max ,cm -1 :3052.43(Ph-H),3023.21(C=C–H),2837.25,1640.81(Ph),1621.35(C=C),1520.35,1462.32 (1,2,3-triazol),1348.48,1220.06,1078.64,821.49,722.32; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 43 N3;Calculated:408.3300;Found:408.3374.

[0073] Compound 5b: white solid, yield 65.50%; mp 66-69℃; [α] 20 D = -34.1 (c 1.0, CHCl3); 1 HNMR(400MHz,CDCl3)δ7.73(d,J=8.8Hz,2H,H Ph ),7.59(s,1H,H 21 ),7.11(d,J=8.2Hz,1H,H,H 14 ), 6.95(s, 2H Ph ), 6.90(d, J = 18.1 Hz, 2H, H 11 H 12 ), 4.37 (d, J = 14.0 Hz, 1H, H 18 ), 4.11(d, J = 14.0 Hz, 1H, H 18 ),3.82(s,3H,H Ph-O-CH3 ),2.95(dd,J=8.9,4.3Hz,2H,H7),2.85–2.74(m,1H,H 15 ),2.17(dd,J=62.6,12.8Hz,2H,H1),1.68–1.16(m,16H),1.09(s,3H,H 19); 13 C NMR (100MHz, CDCl3) δ159.49,146.88,146.61,145.68,134.46,126.97,126.92,124.11,123.87,123.36,120.50,114.18,77. 32,77.00,76.68,61.07,55.28,45.15,38.21,37.97,37.58,36.51,33.39,29.96,25.55,23.92,19.27,18.70,18.48; FT-IRν max ,cm -1 :3309.56,3029.65(Ph-H),3002.52(C=C–H),1665.36(Ph),1655.22(C=C),1589.65,1447.89,1424.25(1,2,3-triazol ),1375.68,1324.23,1307.36,1158.52(CO),1108.63,1057.78,952.37,848.41,564.56; HR-MS(ESI-TOF)m / z[M+H]+;C 29 H 37 N3O;Calculated:444.2937;Found:444.3007.

[0074] Compound 5c: 1H and 1C spectra are shown below. Figure 7 and 8 White solid, yield 88.50%; mp 68-69℃; [α] 20 D = -30.5 (c 1.0, CHCl3); 1 H NMR (400MHz, CDCl3) δ8.21 (td, J=7.6, 1.9Hz, 1H, H 21 ), 7.78 (d, J = 3.7 Hz, 1H, H) 14 ), 7.25–7.11(m,2H,H Ph ), 7.07–6.99(m, 2H Ph ), 6.87 (d, J = 8.2 Hz, 1H, H 11 ),6.81(s,1H,H 12 ), 4.34 (d, J = 14.0 Hz, 1H, H 18 ), 4.05 (d, J = 14.0 Hz, 1H, H 18),2.87(dd,J=8.8,4.2Hz,2H,H7),2.77–2.67(m,1H,H 15 ),2.16(d,J=12.9Hz,1H,H1),2.05–1.98(m,1H,H1),1.22–1.11(m,16H),0.83–0.71(m,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ160.39,157.93,146.59,145.65,134.44,129.13,129.05,127.77,127.73,126.93,124.11,123.86,115.64,115 .42,77.32,77.00,76.68,60.94,44.98,38.25,37.94,37.58,36.38,33.38,29.89,29.67,25.58,23.91,19.27,18.75,18.45; FT-IRν max ,cm -1 :3406.12,3102.86(Ph-H),3051.03(C=C–H),1621.58(Ph),1506.89(C=C),1488.66 (1,2,3-triazol),1355.88,1082.47,869.59,736.56; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 FN3;Calculated:432.2737;Found:432.2810.

[0075] Compound 5d: 1H and 1C spectra are shown below. Figure 9 and 10 White solid, yield 90.00%; mp 66-69℃; [α] 20 D = -31.8 (c 1.0, CHCl3); 1 H NMR (400MHz, CDCl3) δ7.69 (dd, J=9.7, 4.2Hz, 3H, H Ph ),7.58(s,1H,H 21 ), 6.97 (d, J = 8.6 Hz, 3H, H Ph H 14 H 11 ),6.78(s,1H,H 12 ), 4.25 (d, J = 14.0 Hz, 1H, H 18), 3.99 (d, J = 13.9 Hz, 1H, H 18 ),2.89–2.61(m,3H,2H7,H 15 ),2.13(d,J=12.9Hz,1H,H1),1.31(s,4H),1.16–1.06(m,12H),0.89(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ163.61,145.52,134.27,127.29,127.25,127.20,127.17,126.78,123.75,121.10,115.71,115.67,115.49,115 .45,77.32,77.00,76.68,60.91,45.04,38.03,37.81,37.42,36.31,33.25,29.82,25.40,23.80,19.75,19.11,18.51,18.32; FT-IRν max ,cm -1 :3438.96,3140.37(Ph-H),2999.61(C=C–H),1658.59(Ph),1650.48(C=C),1500.26,1490.61 (1,2,3-triazol),1321.26,1128.42,1074.50,935.57,587.62; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 FN3;Calculated:432.2737;Found:432.2809.

[0076] Compound 5e: white solid, yield 75.50%; mp 66-68℃; [α] 20 D = -31.2 (c 1.0, CHCl3); 1 HNMR(400MHz,CDCl3)δ7.72(t,J=1.6Hz,1H,H Ph ), 7.62(d, J = 8.4 Hz, 2H, H Ph H 21 ), 7.24–7.16(m,2H,H Ph ), 7.03 (d, J = 8.2 Hz, 1H, H 14 ), 6.91–6.79(m,2H,H 11 H 12 ), 4.30 (d, J = 14.0 Hz, 1H, H18 ), 4.04 (d, J = 14.0 Hz, 1H, H 18 ),2.91–2.66(m,3H,2H7,H 15 ),2.22–1.94(m,2H,H1),1.28–1.09(m,16H),1.00(s,3H,H 19 ); 13 C NMR (101MHz, CDCl3) δ146.52,145.73,145.69,134.68,134.38,132.37,130.04,127.94,126.91,125.69,124.08,123.88,123.72,121 .66,77.32,77.00,76.68,61.15,45.14,38.19,37.93,37.54,36.48,33.36,29.92,29.65,25.52,23.90,19.24,18.65,18.42; FT-IRν max ,cm -1 :3410.43,3053.62(Ph-H),3011.20(C=C–H),1604.75(Ph),1602.36(C=C),1417.39(1,2,3-t riazol),1402.84,1292.37,1086.28,867.88,736.54,685.69; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 ClN3;Calculated:448.2441;Found:448.2513.

[0077] Compound 5f: pale yellow solid, yield 85.20%; mp 71-74℃; [α] 20 D = -33.3 (c 1.0, CHCl3); 1 H NMR(400MHz,CDCl3)δ7.73–7.65(m,2H,H Ph ), 7.59 (d, J = 1.9 Hz, 1H, H 21 ), 7.31(t, J = 8.4 Hz, 3H, 2H Ph H 14 ), 7.21–7.02(m,1H,H 11 ), 6.91–6.80 (m, 1H, H) 12 ),4.28(dd,J=40.0,13.9Hz,1H,H 18), 4.08–4.00 (m, 1H, H) 18 ),2.79(ddd,J=20.8,11.5,5.6Hz,1H,H 15 ),2.27–1.92(m,2H,H7),1.33–1.01(m,18H),0.94(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ146.55,146.02,145.74,134.41,133.73,129.26,129.00,128.97,126.93,126.89,125.91,124.12,123.91,12 1.34,77.32,77.00,76.68,61.15,46.67,45.13,40.24,38.24,36.51,33.39,29.95,29.68,27.15,25.55,23.92,18.71,1.00; FT-IRν max ,cm -1 :3411.25,3082.21(Ph-H),2918.29(C=C–H),2089.69,1671.18(Ph),1459.81(C=C),1398.56(1,2,3-triaz ol),1358.68,1296.05,1132.31,1039.52,1008.21,889.36,829.56,783.02; HR-MS(ESI-TOF)m / z[M+H]+;C 28 H 34 ClN3; Calculated:448.2441; Found:448.2512.

[0078] Compound 5 g: white solid, yield 80.40%; mp 75-76℃; [α] 20 D = -24.6 (c 1.0, CHCl3); 1 HNMR (400MHz, CDCl3) δ8.07 (d, J = 2.1Hz, 2H, H Ph ),7.58(s,1H,H 21 ),7.56(s,1H,H 14 ), 7.32(d, J = 7.5 Hz, 2H, H Ph ), 7.14–7.09(m,2H,H 11 H 12 ), 4.25 (d, J = 14.0 Hz, 1H, H 18), 4.08 (d, J = 13.9 Hz, 1H, H 18 ),1.35–1.11(m,21H),0.93(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ144.51,137.70,136.34,133.43,131.43,130.60,130.48,129.15,127.65,125.85,125.62,124.59,121.29,121 .23,77.32,77.00,76.68,61.07,46.23,40.30,38.09,37.58,36.24,31.68,31.06,22.18,20.79,19.83,19.51,18.75,18.03; FT-IRν max ,cm -1 :3399.32,3052.56(Ph-H),2921.06(C=C–H),1661.23(Ph),1551.45(C=C),1460.56(1,2,3-tria zol),1449.26,1368.26,1050.94,944.20,856.21,788.41,658.19; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 BrN3; Calculated:492.1936; Found:492.2008.

[0079] Compound 5h: white solid, yield 70.20%; mp 76-77℃; [α] 20 D = -28.7 (c 1.0, CHCl3); 1 HNMR(400MHz,CDCl3)δ7.94–7.86(m,1H,H Ph ), 7.73–7.66(m,1H,H) Ph ), 7.61(d, J = 2.6 Hz, 1H, H 21 ), 7.40–7.20(m,2H,H Ph ,),7.19(d,J=4.1Hz,1H,H 14 ), 7.08–6.80 (m, 2H, H) 11 H 12 ), 4.36–4.17(m,1H,H) 18 ), 4.08–3.99(m,1H,H) 18),2.96–2.66(m,2H,H7),2.26–2.13(m,1H,H 15 ),1.43–1.09(m,18H),0.93(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ145.72,145.64,137.57,134.40,132.78,130.88,130.36,128.61,126.93,125.93,124.20,124.16,122.92,121 .64,77.32,77.00,76.68,61.20,45.18,40.23,38.23,38.00,37.95,37.57,36.53,33.39,29.94,25.53,23.93,18.68,18.45; FT-IRν max ,cm -1 :3354.28,3064.25(Ph-H),2971.44(C=C–H),1605.26(Ph),1601.88(C=C),1430 .22(1,2,3-triazol),1303.66,1108.63,886.06; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 BrN3; Calculated:492.1936; Found:492.2011.

[0080] Compound 5i: white solid, yield 69.50%; mp 56-58℃; [α] 20 D = -24.1 (c 1.0, CHCl3); 1 HNMR(400MHz,CDCl3)δ7.74–7.71(m,2H,H Ph ),7.59(s,1H,H 21 ), 7.30(t, J = 7.6 Hz, 2H, H Ph ), 7.21(d, J = 7.4 Hz, 1H, H Ph ), 7.02(d, J = 8.2 Hz, 1H, H 14 ), 6.89–6.78(m,2H,H) 11 H 12 ), 4.27 (d, J = 14.0 Hz, 1H, H 18 ), 4.02(d, J = 14.0 Hz, 1H, H 18),2.86(dd,J=8.9,4.3Hz,2H,H7),2.77–2.66(m,1H,H 15 ),2.15(d,J=13.0Hz,1H,H1),1.60–1.56(m,1H),1.42–1.08(m,16H),0.99(s,3H,H 19 ); 13 CNMR(100MHz, CDCl3)δ146.95,146.54,145.62,134.38,130.58,128.71,127.94,126.92,126.81,125.67,125.56,124.06,121.36,12 1.22,77.32,77.00,76.68,61.00,45.20,45.07,38.14,37.52,36.43,33.43,33.26,29.91,25.57,23.96,23.81,18.68,18.57; FT-IRν max ,cm -1 :3388.54,3092.66(Ph-H),3054.34(C=C–H),1625.32(Ph),1588.73(C=C),1504.41(1,2,3-triaz ol),1301.07,1299.30,1141.29,1044.28,966.88,880.21,703.02; HR-MS(ESI-TOF)m / z[M+H]+;C 28 H 35 N3;Calculated:414.2831;Found:414.2902.

[0081] Compound 5j: 1H and 1C spectra are shown below. Figure 11 and 12 Pale yellow solid, yield 84.60%; mp 65-66℃; [α] 20 D = -24.9 (c 1.0, CHCl3); 1 H NMR (400MHz, CDCl3) δ7.85 (d, J = 8.1Hz, 2H, H Ph ),7.68(s,1H,H 21 ), 7.58 (d, J = 8.2 Hz, 2H, H Ph ), 7.04 (d, J = 8.2 Hz, 1H, H 14 ), 6.89(dd, J = 8.2, 1.5 Hz, 1H, H 11 ),6.81(s,1H,H 12), 4.34 (d, J = 14.0 Hz, 1H, H 18 ), 4.05 (d, J = 14.0 Hz, 1H, H 18 ),2.93–2.84(m,2H,H7),2.78–2.66(m,1H,H 15 ),2.18(d,J=13.0Hz,1H,H1),2.06–1.96(m,1H,H1),1.37–1.11(m,16H),1.02(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ146.52,145.76,145.69,134.37,134.06,130.01,129.68,126.93,125.79,125.74,125.71,124.11,123.93,122.75 ,122.05,77.32,77.00,76.68,61.20,45.15,38.25,37.95,37.59,36.51,33.39,29.94,29.67,25.54,23.90,19.28,18.70,18.44; FT-IRν max ,cm -1 :3425.10,3028.29(Ph-H),3014.11(C=C–H),1656.12(Ph),1614.87(C=C),1444.58(1,2,3-t riazol),1338.66,1213.33,1109.04,1064.69,968.49,843.98; HR-MS(ESI-TOF)m / z[M+H]+; C 29 H 34 F3N3; Calculated:482.2705; Found:482.2780.

[0082] Compound 5K: pale yellow solid, yield 64.50%; mp 89-90℃; [α] 20 D = -35.8 (c 1.0, CHCl3); 1 H NMR(400MHz, CDCl3)δ8.19–8.14(m,2H,H Ph ), 7.93–7.87(m,2H,H) Ph ),7.78(s,1H,H 21 ), 7.03 (d, J = 8.2 Hz, 1H, H 14 ), 6.91–6.78(m,2H,H) 11 H12 ), 4.36 (d, J = 14.0 Hz, 1H, H 18 ), 4.06 (d, J = 14.0 Hz, 1H, H 18 ),2.93–2.80(m,2H,H7),2.72(dq,J=13.8,6.9Hz,1H,H 15 ),2.17(d,J=13.0Hz,2H,H1),1.36–1.08(m,16H),1.02(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ147.11,146.43,145.72,144.82,136.88,134.28,126.92,126.88,126.05,126.00,124.17,124.05,123.90,122 .82,77.32,77.00,76.68,61.17,45.05,38.22,37.87,37.52,36.43,33.32,29.88,29.62,25.49,23.86,19.22,18.66,18.38; FT-IRν max ,cm -1 :3431.05,3144.20(Ph-H),2996.81(C=C–H),2108.48,1665.11(Ph),1597.80(C=C), 1465.98(1,2,3-triazol),1337.54,1217.69,998.58; HR-MS(ESI-TOF)m / z[M+H]+; C 28 H 34 N4O2; Calculated:459.2682; Found:459.2759.

[0083] Compound 5l: white solid, yield 66.00%; mp 62-63℃; [α] 20 D = -28.6 (c 1.0, CHCl3); 1 HNMR (400MHz, CDCl3) δ7.61 (d, J = 8.1Hz, 2H, H Ph ),7.55(s,1H,H 21 ),7.12(d,J=8.0Hz,2H,H Ph ),7.02(d,J=8.2Hz,1H,H7),6.89–6.77(m,2H,H 11 H 12), 4.26 (d, J = 14.0 Hz, 1H, H 18 ), 4.01(d, J = 14.0 Hz, 1H, H 18 ),2.85(dd,J=8.9,4.3Hz,2H,H7),2.76–2.67(m,1H,H 15 ),2.27(s,3H,H Ph-CH3 ),2.15(d,J=12.9Hz,2H,H1),1.65–1.06(m,16H),0.99(s,3H,H 19 ); 13 C NMR (100MHz, CDCl3) δ147.03,146.57,145.62,137.74,134.40,129.39,127.79,126.95,126.80,125.63,125.46,124.06,121.04,120.85 ,77.32,77.00,76.68,60.98,45.26,38.14,37.53,36.46,33.45,33.26,29.92,25.58,23.98,23.79,21.25,21.13,18.70,18.55; FT-IRν max ,cm -1 :3441.11,3149.36,3052.56(Ph-H),3009.55(C=C–H),2188.59,1617.58(Ph),1607.04(C=C) ,1456.89(1,2,3-triazol),1389.87,1302.44,998.58,656.42; HR-MS(ESI-TOF)m / z[M+H]+;C 29 H 37 N3;Calculated:428.2987;Found:428.3060.

[0084] Example 5: In vitro antitumor activity test

[0085] The MTT assay was used to test the in vitro growth and proliferation inhibitory activity of the target compound against three cell lines: human cervical cancer cells (HeLa), human breast cancer cells (MCF-7), and human renal epithelial cells (HEK-293T). All cells were cultured in DMEM medium (containing 10% bovine serum and 0.1 g / L penicillin G + 0.1 g / L streptomycin sulfate) at 37°C under a humid atmosphere of 5% CO2. The target compound was dissolved in sterile DMSO (Sigma) at a concentration of 10 mg / mL and diluted with culture medium according to the intended use concentration.

[0086] Cell viability assessment: Cells in the logarithmic growth phase were inoculated at approximately 3 × 10⁻⁶ cells per cell. 4 Cells were seeded at a density of 200 μL / mL in 96-well plates and incubated in a CO2 incubator for 24 h. Then, different concentrations of the target compound were added to the test wells, with 6 parallel wells for each concentration gradient. An equal volume of DMSO was used as a blank control. The cells were incubated in a CO2 incubator for another 48 h. The cells were washed with sterile PBS (phosphate buffer saline), and then 190 μL of culture medium and 10 μL of MTT (5 mg / mL) solution were added to each well. The cells were incubated in a CO2 incubator for another 4 h. The supernatant was discarded, and 200 μL of DMSO was added. The cells were shaken on a shaker for 10 min to dissolve the formed purple precipitate. The OD value was then measured at a wavelength of 490 nm using a microplate reader. The test results are shown in Table 1.

[0087] Table 1. In vitro antiproliferative activity of the dehydrorosinic acid 1,2,3-triazole derivative of the present invention [IC50] 50 / (μmol·L -1 )]

[0088]

[0089] As shown in Table 1, compound 5d exhibits excellent inhibitory activity against all tested tumor cell lines, such as the inhibitory activity against human cervical cancer cells (HeLa) with an IC50 value. 50 The value is 22.1 μmol·L -1 Inhibitory activity of human breast cancer cells (MCF-7) IC50 50 The value is 23.2 μmol·L -1 However, it showed virtually no inhibitory activity against normal human renal epithelial cells (HEK-293T) (IC50). 50 Value > 100 μmol·L -1 Compound 5j showed inhibitory activity against all tested tumor cell lines, such as an inhibitory activity (IC50) against human cervical cancer cells (HeLa). 50 The value is 26.5 μmol·L. -1 Inhibitory activity of human breast cancer cells (MCF-7) IC50 50 The value is 32.7 μmol·L -1 However, it showed virtually no inhibitory activity against normal human renal epithelial cells (HEK-293T) (IC50). 50 Value > 100 μmol·L -1 Compound 5c showed inhibitory activity against all tested tumor cell lines, such as an inhibitory activity of IC50 against human cervical cancer cells (HeLa).50 The value is 34.5 μmol·L -1 Inhibitory activity of human breast cancer cells (MCF-7) IC50 50 The value is 42.4 μmol·L -1 However, it showed virtually no inhibitory activity against normal human renal epithelial cells (HEK-293T) (IC50). 50 Value > 100 μmol·L -1 Therefore, the dehydrorosinic acid derivatives of this invention have certain inhibitory activity on the growth and proliferation of tumor cells, while causing less damage to normal cells. Among them, the antitumor effects of 5d and 5j are significantly better than those of dehydrorosinic acid and cisplatin, providing a research basis and lead compounds for new drug screening, and have good application prospects.

[0090] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Any other modifications or equivalent substitutions made by those skilled in the art to the technical solutions of the present invention, as long as they do not depart from the spirit and scope of the technical solutions of the present invention, should be covered within the scope of the claims of the present invention.

Claims

1. A dehydrorosinic acid 1,2,3-triazole derivative, characterized in that, Its chemical structural formula is as follows: In the formula: R represents an alkyl or substituted phenyl group.

2. The dehydrorosinic acid 1,2,3-triazole derivative according to claim 1, characterized in that, In the formula, the alkyl group is n-pentyl, 3-bromopropyl, n-butyl, 5-hydroxypentyl, 1-methylbutyl, 4-chlorobutyl, or 3-cyanopropyl; The substituted phenyl group is 4-methoxyphenyl, 2-fluorophenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2-bromophenyl, 3-bromophenyl, phenyl, 4-trifluoromethylphenyl, 4-nitrophenyl, or 4-methylphenyl.

3. A method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 1 or 2, characterized in that, Includes the following steps: (1) In the first organic solvent, dehydrorosin acid reacts under the action of a reducing agent. After the reaction is completed, an alkaline aqueous solution is added to quench the reaction. Then, the mixture is extracted and separated to obtain compound 2 with reduced carboxyl groups. (2) Compound 2 reacts with p-methylbenzenesulfonyl chloride in an alkaline solution. After the reaction is completed, an acidic buffer solution is added to quench the reaction. Then, the mixture is extracted and separated to obtain compound 3 with the hydroxyl group protected. (3) In the second organic solvent, compound 3 reacts with sodium azide under high temperature conditions. After the reaction is completed, an acidic aqueous solution is added to quench the reaction, and then the mixture is extracted and separated to obtain azide compound 4. (4) Compound 4 and copper salt react with alkyne compound in a polar alkaline solvent. After the reaction is completed, water is added to quench the reaction, followed by extraction and separation to obtain the target compound.

4. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, The molar ratio of dehydrorosin acid and reducing agent in step (1) is 1:2-6; The first organic solvent is one or more of diethyl ether, tetrahydrofuran, 1,4-dioxane, methyl tert-butyl ether and acetonitrile; The reducing agent is one or more of lithium aluminum hydride, sodium borohydride and sodium cyanoborohydride; The alkaline aqueous solution is a sodium hydroxide aqueous solution with a mass concentration of 10-20%, a sodium bicarbonate aqueous solution with a mass concentration of 20-40%, or a sodium carbonate aqueous solution with a mass concentration of 15-25%.

5. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, The molar ratio of compound 2 and p-methylbenzenesulfonyl chloride in step (2) is 1:2-7; The alkaline solution is one or more of triethylamine, triethanolamine, and pyridine; The acidic buffer solution is one or more of the following: saturated ammonium chloride solution, acetic acid-sodium acetate solution, and citric acid-sodium citrate solution.

6. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, In step (3), the molar ratio of compound 3 to sodium azide is 1:2-6; the temperature of the high-temperature condition is 40℃-100℃.

7. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, In step (3), the second organic solvent is one or more of hexamethylphosphoric triamine, N,N-dimethylformamide and dimethyl sulfoxide; The acidic aqueous solution is a 5-15% sulfuric acid aqueous solution, a 5-15% hydrochloric acid aqueous solution, or a 5-15% nitric acid aqueous solution.

8. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, In step (4), the molar ratio of compound 4, copper salt, and alkyne compound is 1:0.2-1:1-3.

9. The method for preparing the dehydrorosinic acid 1,2,3-triazole derivative according to claim 3, characterized in that, The copper salt mentioned in step (4) is one or more of copper sulfate, copper acetate, copper chloride, copper nitrate and copper carbonate; The alkali is one or more of sodium carbonate, sodium bicarbonate, sodium sulfite, sodium bisulfite, and sodium ascorbate; The polar solvent is one or more of water-methanol, water-ethanol, water-isopropanol, water-n-butanol, and water-tert-butanol.

10. The use of the dehydrorosinic acid 1,2,3-triazole derivative of claim 1 or 2 in the preparation of a medicament for treating cancer.

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