Synthesis method and anti-tumor application of C1-triazole oridonin derivative

Abstract

The invention discloses a synthetic method of a C1-triazole oridonin derivative and an anti-tumor application of the C1-triazole oridonin derivative. The invention provides a synthetic method of the C1-triazole oridonin derivative, which comprises the following steps: carrying out four-step reaction to obtain an oridonin analogue containing an azide group at the C1 site, and catalyzing by cuprous iodide to obtain the C1-triazole oridonin derivative with the azide group at the C1 site, thereby obtaining the C1-triazole oridonin derivative with the azide group at the C1 site and the oridonin analogue with the azide group at the C1 site. According to the method, alkyne compounds containing different substituent groups react with C1-triazole oridonin to synthesize the C1-triazole oridonin derivative, the method is mild in condition and rapid in reaction, large-scale preparation of the C1-triazole oridonin derivative can be achieved, and the method is suitable for large-scale production of the C1-triazole oridonin derivative. Meanwhile, the C1-triazole oridonin derivative synthesized by the invention can be used for effectively inhibiting the proliferation activity of tumor cells and has a relatively good inhibition effect on MCF-7 cells and A549 cells.

Description

Technical Field

[0001] The present invention relates to the technical field of organic synthesis, in particular to a synthesis method and anti-tumor application of C1-triazole oridonin derivatives. Background Art

[0002] Oridonin is a natural product of the ent-kaurane tetracyclic diterpenoids extracted from the plant Oridonium rubescens. Numerous studies have shown that Oridonin has a variety of pharmacological activities, such as anti-tumor, antibacterial, anti-inflammatory, immunomodulatory, and neuroprotective activities. Among these pharmacological activities, the anti-tumor activity of Oridonin is particularly prominent and has become a hot topic of research at home and abroad. Decades of research have found that Oridonin is effective against cancers such as esophageal cancer, breast cancer, non-small cell lung cancer, colon cancer, oral cancer, liver cancer, prostate cancer, and pancreatic cancer. Oridonin has good safety and biological activity, but its moderate potency and poor water solubility have greatly reduced the effectiveness of Oridonin in clinical applications. However, as an unmodified natural product, Oridonin has disadvantages such as poor water solubility and low oral bioavailability. Structural modification can not only overcome the disadvantages of Oridonin's low water solubility, but also improve its anti-tumor activity. Triazole is a common group in anti-tumor drugs, which can produce anti-tumor effects in various ways, such as inhibiting tumor cell proliferation and inducing tumor cell apoptosis.

[0003] In the prior art, invention CN 118530249A uses oridonin as a lead compound to design and synthesize oridonin isothiocyanate and thiocarbamate hydrogen sulfide donor derivatives. These can be used to treat a variety of tumor cells, including human breast cancer cells MCF-7, human lung adenocarcinoma cells A549, human liver cancer cells Bel-7402, human liver cancer cells HepG2, and human pancreatic cancer cells Panc-1 and MIA-PaCa-2. However, these derivatives are toxic to humans, and the chemical reactions are complex and harsh, with a low reaction rate. Invention CN 114874172B prepares a novel oridonin derivative, which primarily acts as an anti-inflammatory agent and does not play a key role in cancer prevention. Triazole is a common group in antitumor drugs that can produce antitumor effects through various means, such as inhibiting tumor cell proliferation and inducing tumor cell apoptosis. Currently, there are no related experiments combining triazole, a common group in antitumor drugs, with oridonin.

[0004] Therefore, there is an urgent need for a method for synthesizing Rubescensine A derivatives with good activity in a mild, rapid and simple manner. Summary of the Invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a synthesis method and anti-tumor application of a C1-triazole oridonin derivative, so as to at least achieve the synthesis of an oridonin derivative with good activity and water solubility using fewer steps and a shorter time.

[0006] The object of the present invention is achieved through the following technical solutions:

[0007] C1-triazole Rubescensine A derivatives as shown in formula (I) or formula (II):

[0008]

[0009] Where R is selected from One of the following;

[0010]

[0011] Wherein R1 is selected from One of them.

[0012] On the one hand, the synthesis steps of the C1-triazole oridonin derivative include: dissolving C1-azido oridonin C in an organic solvent, adding a catalyst, a ligand and an organic base; adding an acetylenic compound, heating to react, and extracting to obtain the C1-triazole oridonin derivative I;

[0013] The structural formula of the C1-azidorubescensine A c is:

[0014] Preferably, the organic solvent is toluene, benzene, tetrahydrofuran or acetonitrile;

[0015] Preferably, the catalyst is CuI, CuBr or CuCl;

[0016] Preferably, the ligand is triphenylphosphine, bipyridine, 1,10-phenanthroline or a derivative thereof;

[0017] Preferably, the organic base is N,N-diisopropylethylamine, triethylamine, DBU or DIPEA;

[0018] Specifically, the heating reaction temperature is 80-85°C;

[0019] Specifically, the mass ratio of the C1-triazole Rubescensine A c to the alkyne is 1:10-12.

[0020] On the other hand, the deketone reaction step of the C1-triazole oridonin derivative comprises: dissolving the C1-triazole oridonin derivative I in an organic solvent, adding an acid catalyst, and reacting to obtain the C1-triazole oridonin derivative II;

[0021] Specifically, the organic solvent is tetrahydrofuran (THF), dichloromethane (DCM), ether or ethyl acetate;

[0022] Specifically, the acid catalyst is concentrated hydrochloric acid, sulfuric acid, trifluoroacetic acid, p-toluenesulfonic acid or aluminum chloride;

[0023] Preferably, the reaction temperature is 15-25°C.

[0024] In another aspect, a method for synthesizing C1-triazole Rubescensine A comprises the following steps:

[0025] S1: dissolving oridonin in an organic solvent, adding an acid catalyst, and reacting to obtain oridonin with a protective group; dissolving the oridonin with a protective group in an inert organic solvent, adding an organic base and a sulfonylating agent, and reacting to obtain 1-methoxysulfonyl-substituted oridonin a with a protective group, as shown in the following reaction formula:

[0026]

[0027] S2: dissolving the 1-methylsulfonyl-substituted protected Rubescensine A a in a polar aprotic solvent, adding an inorganic base and a halide salt to react to obtain a C1 halogenated Rubescensine A analogue b, and the reaction is as follows:

[0028]

[0029] S3: dissolving the C1-halogenated Rubescensine A analogue b in an organic solvent, adding an azidation reagent and a catalyst to react to obtain the C1-triazole Rubescensine A c, and the reaction is as follows:

[0030]

[0031] Preferably, the organic solvent in step S1 is 2,2-dimethoxypropane;

[0032] Preferably, the acid catalyst in step S1 is p-toluenesulfonic acid, sulfuric acid, trifluoroacetic acid, camphorsulfonic acid or Lewis acid;

[0033] Preferably, the inert organic solvent in step S1 is dichloromethane, tetrahydrofuran, diethyl ether or ethyl acetate;

[0034] Preferably, the sulfonylating agent in step S1 is methanesulfonyl chloride, p-toluenesulfonyl chloride or trifluoromethanesulfonic anhydride;

[0035] Preferably, the polar aprotic solvent in step S2 is N,N-dimethylformamide, dimethyl sulfoxide, N-methylpyrrolidone or hexamethylphosphoramide;

[0036] Preferably, the inorganic base in step S2 is lithium carbonate, sodium carbonate, potassium carbonate or cesium carbonate;

[0037] Preferably, the halide salt in step S2 is lithium bromide, sodium bromide, potassium bromide, lithium chloride or sodium iodide;

[0038] Preferably, the organic solvent in step S3 is acetonitrile, N,N-dimethylformamide, tetrahydrofuran or dimethyl sulfoxide;

[0039] Preferably, the azidation reagent in step S3 is trimethylsilyl azide, sodium azide or lithium azide;

[0040] Preferably, the catalyst in step S3 is tetrabutylammonium fluoride, cesium fluoride or potassium fluoride;

[0041] Specifically, the temperature of the reaction for obtaining Rubescensine A with a protecting group in step S1 is 15 to 25 ° C, the reaction time is 1 hour, and the extraction step is: after quenching with water, extracting with ethyl acetate, washing the organic phase with saturated brine, adding anhydrous sodium sulfate and filtering, and removing volatile substances by rotary evaporation under reduced pressure; the reaction temperature of Rubescensine A a in step S1 is 0 to 4 ° C, the reaction time is 1 hour, the extraction step is: after quenching with water, extracting with ethyl acetate 3 times, washing the organic phase with saturated brine, adding anhydrous sodium sulfate and filtering, removing volatile substances by rotary evaporation under reduced pressure, and separating the residue by column chromatography.

[0042] The reaction temperature in step S2 is 90° C., the reaction time is 2.5 h, and the extraction step is as follows: quenching with water, extracting with ethyl acetate, collecting the organic phase, washing with saturated brine, filtering with anhydrous sodium sulfate, removing volatile substances by reduced pressure rotary evaporation, and separating the residue by column chromatography.

[0043] The reaction temperature in step S3 is 80° C., the reaction time is 2 h, and the extraction step is as follows: quenching with water, extracting with ethyl acetate, collecting the organic phase, washing with saturated brine, filtering with anhydrous sodium sulfate, removing volatile substances by reduced pressure rotary evaporation, and separating the residue by column chromatography.

[0044] In another aspect, the C1-triazole oridonin derivative is used to prepare an anticancer drug.

[0045] The beneficial effects of the present invention are:

[0046] The present invention uses copper catalysis to introduce an azide group at the C1 position of oridonin without changing the other structures of oridonin. This reaction is not only simple and efficient, but also mild and rapid, enabling the large-scale preparation of C1-triazole oridonin derivatives.

[0047] The C1-triazole oridonin derivative of the present invention has good water solubility, can effectively inhibit the proliferation activity of tumor cells, has a good inhibitory effect on MCF-7 cells and A549 cells, and has the prospect of being used in the treatment of breast cancer and lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The reaction equation for preparing the substrate a of Example 1C1-methylsulfonyl ester Rubescensine A is as follows;

[0049] Figure 2 The reaction equation for preparing the substrate b of Example 1C1-Russiconazole is as follows;

[0050] Figure 3 The reaction equation for preparing the substrate c of Example 1C1-Azidorubescensine A is as follows;

[0051] Figure 4 This is the reaction equation for the synthesis of the triazole Rubescensine A derivative in Example 3C1;

[0052] Figure 5 This is the deketone reaction equation of the 1-triazole Rubescensine A derivative of Experimental Example 3C. DETAILED DESCRIPTION

[0053] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings, but the protection scope of the present invention is not limited to the following.

[0054] Example 1

[0055] The synthesis method of C1-triazole Rubescensine A comprises the following steps:

[0056] ① Oridonin (4.11mmol, 1.50g) was used as the raw material and dissolved in 10mL of 2,2-dimethoxypropane. After stirring, p-toluenesulfonic acid (5.0mmol, 861mg) was added and reacted at room temperature for 1 hour. When the solution turned from turbid to clear, the reaction was complete. After quenching with water, the mixture was extracted with ethyl acetate (30mL). The organic phase was collected and washed with saturated brine three times. Anhydrous sodium sulfate (Na2SO 4) , filtered, and volatile substances were removed by rotary evaporation under reduced pressure to obtain Rubescensine A with a protective group (propylidene structure). The reacted Rubescensine A was placed in an ice-water bath, and dichloromethane (15 mL) was used as a solvent. Stirring was performed, and triethylamine (24.66 mmol, 3.42 μL) and methanesulfonyl chloride (12.33 mmol, 952.66 μL) were added. The mixture was reacted for 1 hour. After quenching with water, the mixture was extracted three times with ethyl acetate (45 mL). The organic phase was collected, washed three times with saturated brine, and anhydrous sodium sulfate (Na2SO 4)，Filter, remove volatile substances by pressure rotary evaporation, and separate the residue by column chromatography (petroleum ether: ethyl acetate) to obtain 1-methoxysulfonyl substituted oridonin A with a protective group. The preparation reaction is as follows: Figure 1 shown.

[0057] ② The obtained a was dissolved in N,N-dimethylformamide (DMF 20mL), LiCO3 (6.64mmol, 490mg), LiBr (6.64mmol, 575mg), stirred, and placed in a 90℃ oil bath to react for 2.5 hours. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate (60mL). The organic phase was collected, washed with saturated brine three times, and then anhydrous sodium sulfate (Na2SO 4) , filtered, and volatile substances were removed by vacuum rotary evaporation. The residue was separated by column chromatography (petroleum ether: ethyl acetate) to obtain a C1-position bromo-substituted oridonin analog b. The preparation reaction is as follows: Figure 2 shown.

[0058] ③ The obtained Rubescensine A analogue b was dissolved in acetonitrile (20 mL), stirred, and trimethylsilane (TMSN3, 3.21 mmol, 422 μL) was added, and tetrabutylammonium fluoride (TBAF, 3.21 mmol, 3.18 mL) was added. The mixture was placed in an 80°C oil bath to react for 2 hours, quenched with water, and extracted with ethyl acetate (60 mL). The organic phase was collected, washed with saturated brine three times, and then anhydrous sodium sulfate (Na2SO 4) Filter, remove volatile substances by rotary evaporation under reduced pressure, and separate the residue by column chromatography (petroleum ether: ethyl acetate) to obtain Rubescensine A analogue c with azide substitution at C1 position. Figure 3 shown.

[0059] Example 2

[0060] Catalytic experimental conditions screening:

[0061] Toluene (1 mL) was used as the solvent. Triphenylphosphine (0.138 mmol, 4 mg) and N,N-diisopropylethylamine (0.207 mol, 36 μL) were added, and the substrate-to-alkyne equivalent ratio was increased to 10-fold. The reaction was carried out at 90°C for 1 hour. TLC analysis revealed that the oridonin analog had reacted completely, with only a single spot formed, for a yield of 70%. The product was isolated and identified by nuclear magnetic resonance (NMR) as the target product.

[0062] Example 3

[0063] The synthesis method of a C1-triazole oridonin derivative comprises the following steps:

[0064] Dissolve c (0.069 mmol, 30 mg) in 1 mL of toluene, add catalyst CuI (0.207 mmol, 4 mg), triphenylphosphine (0.138 mmol, 4 mg), and N,N-diisopropylethylamine (0.207 mol, 36 μL). Add 1a-6a separately, stir, and react at 80°C for 0.5 h. Monitor the reaction progress by TLC until the reaction is complete. Quench with water, extract with ethyl acetate (5 mL), collect the organic phase, wash with saturated brine three times, and then add anhydrous sodium sulfate (Na2SO 4) Dry, filter, and remove volatile substances by vacuum rotary evaporation. The residue is separated by column chromatography (petroleum ether: ethyl acetate) to obtain the target product. The properties and yields are shown in the compound characterization section, and the synthetic reaction is shown in Figure 4 As mentioned above, the reaction is a single addition to the C1 position of Rubescensine A, and the product is initially 1 H and 13 The structure was identified by C-NMR data.

[0065] The deketoneization method of a C1-triazole oridonin derivative comprises the following steps:

[0066] 1b-5b was dissolved in THF (1.5 mL), concentrated hydrochloric acid (1.5 mmol, 0.5 mL) was added, stirred, and reacted at room temperature for 0.5 h. The reaction progress was monitored by TLC until the reaction was completed. Water was added to quench the reaction, and the mixture was extracted with ethyl acetate (5 mL). The organic phase was collected and washed with saturated brine three times, and then anhydrous sodium sulfate (Na2SO 4) Dry, filter, and remove volatile substances by vacuum rotary evaporation. The residue is separated by column chromatography (petroleum ether: ethyl acetate) to obtain the target product. Figure 5 shown.

[0067] Structural characterization of synthesized compounds:

[0068] 1H NMR(600MHz,CDCl3)δ6.16–6.15(m,1H),5.76(d,J=11.8Hz,1H),5.55(t,J=0.8Hz,1H),4.79(d,J=1.5Hz,1H),4.24(dd,J=9.9,1.7Hz,1H),4.05(dd,J=9.9,1.4Hz,1H),3.90(dd,J=11.8,7.5Hz,1H),3.48(dd,J=11.6,5.5Hz,1H),3.06(d,J=8.8Hz,1H),2.53(dt,J=14.0,8.9Hz,1H),2.01–1.92(m,1H),1.80–1.67(m,3H),1.66–1.65(m,3H),1.64–1.60(m,1H),1.45(dt,J=13.8,3.6Hz,1H),1.34(s,3H),1.29–1.23(m,3H),1.17(s,3H),1.16(s,3H),1.03(s,1H).

[0069] 1 H NMR(600MHz,CDCl3)δ6.16(s,1H),5.81(d,J=11.8Hz,1H),5.57(d,J=1.1Hz,1H),4.75(d,J=1.8Hz,1H),4.59(dd,J=11.9,5.7Hz,1H),4.16–4.12(m,2H),3.92(dd,J=11.8,7.5Hz,1H),3.06(d,J=10.1Hz,1H),2.99(s,3H),2.53(ddd,J=13.6,8.0,4.4Hz,1H),2.08(dddt,J=8.7,5.8,4.1,3.0Hz,1H),1.91–1.69(m,6H),1.63(s,3H),1.52(dt,J=14.2,3.6Hz,1H),1.37(dd,J=7.6,1.4Hz,1H),1.33(s,3H),1.19(s,3H),1.18(s,3H).

[0070] 1H NMR(600MHz,CDCl3)δ6.19(s,1H),6.08(d,J=11.5Hz,1H),5.57(d,J=1.1Hz,1H),4.72(d,J=1.9Hz,1H),4.12(dd,J=9.4,2.0Hz,1H),4.03(t,J=3.0Hz,1H),3.92(dd,J=11.5,6.2Hz,1H),3.83(dd,J=9.4,2.0Hz,1H),3.07(d,J=10.8Hz,1H),2.50(dt,J=14.0,8.9Hz,1H),2.09(ddd,J=13.0,5.7,1.9Hz,1H),2.04–1.98(m,1H),1.90(ddd,J=11.7,5.3,3.1Hz,2H),1.83–1.78(m,1H),1.74(ddd,J=13.8,11.0,7.4Hz,1H),1.67–1.63(m,1H),1.62(s,3H),1.60–1.58(m,1H),1.37(dt,J=14.9,3.0Hz,1H),1.33(s,3H),1.22(s,3H),1.19(s,3H).

[0071] 1 H NMR(600MHz,CDCl3)δ6.16(s,1H),5.77(d,J=11.9Hz,1H),5.57(s,1H),4.76(s,1H),4.10(d,J=11.7Hz,1H),3.98(d,J=8.4Hz,1H),3.88(dd,J=11.9,7.6Hz,1H),3.22(dd,J=12.3,5.4Hz,1H),3.07(d,J=9.2Hz,1H),2.56(d,J=22.8Hz,1H),1.96–1.89(m,1H),1.87–1.83(m,1H),1.77–1.73(m,1H),1.72–1.65(m,3H),1.64(s,3H),1.55(t,J=3.4Hz,1H),1.33(s,3H),1.28(dd,J=7.3,2.0Hz,2H),1.19(s,3H),1.15(s,3H).

[0072] 1H NMR(600MHz,CDCl3)δ7.82(d,J=7.7Hz,2H),7.60–7.56(m,1H),7.43(t,J=7.5Hz,2H),7.35(t,J=7.4Hz,1H),6.16(s,1H),6.05(d,J=11.5Hz,1H),5.53(s,1H),4.58(s,1H),4.56–4.50(m,2H),4.33(d,J=9.7Hz,1H),3.97(dd,J=11.7,7.0Hz,1H),2.96(d,J=9.2Hz,1H),2.22(dt,J=14.6,8.8Hz,1H),2.10(qd,J=13.6,3.1Hz,1H),2.04–1.99(m,1H),1.73–1.66(m,2H),1.65–1.62(m,1H),1.60(s,3H),1.56–1.44(m,3H),1.31(s,3H),1.29(s,3H),1.26(s,3H),1.03(qd,J=13.1,8.4Hz,1H). 13 C NMR(151MHz,CDCl3)δ205.39,150.55,147.98,130.23,129.08,128.61,125.76,120.55,119.52,101.12,94.59,72.82,70.15,65.97,63.99,60.35,56.29,50.72,41.19,40.33,40.17,34.15,33.23,30.27,29.95,29.86,25.51,22.61,17.12.

[0073] 1H NMR(600MHz,CDCl3)δ7.75(d,J=8.1Hz,2H),7.53(s,1H),7.46(d,J=8.8Hz,2H),6.16(s,1H),6.05(d,J=11.6Hz,1H),5.54(s,1H),4.58(s,1H),4.55–4.50(m,2H),4.34(d,J=9.7Hz,1H),3.97(dd,J=11.7,7.0Hz,1H),2.96(d,J=9.2Hz,1H),2.25–2.18(m,1H),2.11(qd,J=13.3,2.9Hz,1H),2.02(dd,J=13.4,5.9Hz,1H),1.72–1.63(m,4H),1.61(s,3H),1.52–1.49(m,2H),1.35(d,J=1.1Hz,9H),1.32(s,3H),1.30(s,3H),1.26(s,3H). 13 C NMR(151MHz,CDCl3)δ205.40,151.88,150.56,147.97,127.40,126.02,125.50,120.60,119.21,101.14,94.60,72.83,70.17,65.98,64.04,60.38,56.32,50.74,41.25,40.39,40.19,34.86,34.20,33.26,31.42,30.29,29.99,29.85,25.51,22.67,17.08.

[0074] 1H NMR(600MHz,CDCl3)δ7.02(s,1H),6.14(s,1H),6.00(d,J=11.7Hz,1H),5.53(s,1H),4.59(d,J=2.0Hz,1H),4.46–4.42(m,2H),4.23(d,J=9.7Hz,1H),3.94(dd,J=11.8,7.1Hz,1H),2.96(d,J=9.2Hz,1H),2.25(dt,J=14.6,8.8Hz,1H),2.00–1.94(m,2H),1.89(tt,J=8.7,5.0Hz,1H),1.69(d,J=8.2Hz,1H),1.60(s,5H),1.53(ddd,J=14.4,12.0,7.7Hz,1H),1.46(d,J=7.2Hz,1H),1.42(td,J=14.3,3.5Hz,1H),1.31(s,4H),1.25(s,3H),1.23(s,3H),0.96–0.93(m,2H),0.85–0.82(m,2H). 13 C NMR(151MHz,CDCl3)δ205.38,150.60,150.51,120.48,119.74,101.08,94.60,72.82,70.16,65.62,64.01,60.27,56.30,50.80,41.10,40.34,40.19,34.11,33.25,30.27,30.03,29.71,25.52,22.59,16.93,8.06,8.03,6.80.

[0075] 1H NMR(600MHz,CDCl3)δ8.03(d,J=7.8Hz,2H),7.57(t,J=7.4Hz,1H),7.50(s,1H),7.44(t,J=7.7Hz,2H),6.14(s,1H),6.02(d,J=11.7Hz,1H),5.52(s,1H),5.47–5.40(m,2H),4.55(d,J=1.8Hz,1H),4.50–4.44(m,2H),4.23(d,J=9.8Hz,1H),3.94(dd,J=11.7,7.0Hz,1H),2.93(d,J=9.2Hz,1H),2.14(dt,J=14.3,8.7Hz,1H),2.06(qd,J=14.2,3.6Hz,1H),1.97(dd,J=13.3,6.0Hz,1H),1.60(d,J=11.5Hz,5H),1.52–1.46(m,2H),1.42(td,J=13.8,3.2Hz,1H),1.30(s,3H),1.26(s,3H),1.23(s,4H),1.18(td,J=12.5,5.7Hz,1H),0.91(qd,J=13.4,8.6Hz,1H). 13 C NMR(151MHz,CDCl3)δ205.35,166.58,150.50,143.23,133.48,129.86,129.77,128.59,124.31,120.56,101.10,94.53,72.77,70.11,66.04,63.90,60.34,58.13,56.24,50.63,41.21,40.29,40.12,34.13,33.19,30.26,29.83,29.80,25.53,22.58,17.02.

[0076] 1H NMR(600MHz,CDCl3)δ7.90(s,1H),6.15(s,1H),6.02(d,J=11.6Hz,1H),5.54(s,1H),4.59(dd,J=13.2,4.4Hz,1H),4.56(s,1H),4.49(d,J=9.6Hz,1H),4.42(q,J=7.1Hz,2H),4.18(d,J=9.6Hz,1H),3.95(dd,J=11.7,7.1Hz,1H),2.96(d,J=9.1Hz,1H),2.23(dt,J=14.5,8.8Hz,1H),2.04(dtd,J=26.4,13.2,4.4Hz,2H),1.64(td,J=12.2,3.4Hz,2H),1.59(s,3H),1.57–1.51(m,1H),1.50(d,J=7.2Hz,1H),1.41(t,J=7.2Hz,3H),1.31(s,3H),1.27(s,3H),1.25(s,3H),1.23–1.19(m,1H),0.97–0.90(m,1H),0.86(td,J=16.0,10.3Hz,1H).13C NMR(151MHz,CDCl3)δ205.24,160.67,150.39,140.63,127.43,120.74,101.17,94.56,72.74,70.09,66.32,63.68,61.69,60.26,56.18,50.55,41.05,40.18,40.11,34.11,33.15,30.24,29.90,29.84,25.50,22.54,17.24,14.41.

[0077] 1H NMR(600MHz,CDCl3)δ8.89(d,J=243.4Hz,2H),8.20(d,J=7.8Hz,1H),7.69–7.65(m,1H),7.48–7.39(m,1H),6.16(s,1H),6.05(d,J=11.7Hz,1H),5.54(s,1H),4.59(d,J=1.8Hz,1H),4.58–4.53(m,2H),4.32(d,J=9.7Hz,1H),3.97(dd,J=11.7,7.0Hz,1H),2.96(d,J=9.2Hz,1H),2.24(dt,J=14.6,8.8Hz,1H),2.11(qd,J=13.6,3.0Hz,1H),2.06–2.01(m,1H),1.85(s,1H),1.72–1.62(m,2H),1.60(s,3H),1.57–1.44(m,3H),1.31(s,3H),1.30(s,3H),1.26(s,3H),1.04(qd,J=13.1,8.5Hz,1H).13C NMR(151MHz,CDCl3)δ216.30,147.86,130.57,128.96,128.32,125.76,120.69,101.34,94.47,72.92,72.21,64.42,63.01,59.03,57.32,56.46,52.57,50.24,39.72,39.12,36.19,33.57,33.36,30.38,30.12,25.21,25.04,22.45,19.19.

[0078] 1H NMR(600MHz,DMSO)δ8.52(s,1H),7.85(d,J=7.7Hz,2H),7.44(t,J=7.6Hz,2H),7.33(t,J=7.4Hz,1H),6.94(s,1H),6.28(d,J=9.8Hz,1H),6.19(s,1H),5.98(s,1H),5.56(s,1H),4.80(dd,J=12.9,4.1Hz,1H),4.63(s,1H),4.53(d,J=10.2Hz,1H),4.16(d,J=10.2Hz,1H),3.60(dd,J=9.9,6.0Hz,1H),2.84(d,J=9.8Hz,1H),2.21(dd,J=12.9,6.3Hz,1H),2.12(dtd,J=26.9,13.9,5.4Hz,3H),1.53(d,J=6.4Hz,1H),1.42(p,J=6.8Hz,1H),1.32–1.26(m,2H),1.25(s,1H),1.18(s,3H),1.07(s,3H),0.83–0.75(m,1H).

[0079] 1 H NMR(600MHz,DMSO)δ8.45(s,1H),8.29(s,2H),7.76(d,J=8.0Hz,2H),7.68(ddd,J=28.4,5.8,2.8Hz,2H),7.45(d,J=7.8Hz,2H),6.93(s,1H),6.28(dd,J=9.8,2.8Hz,1H),6.21(s,1H),5.98(s,1H),5.55(s,1H),4.78(dd,J=13.1,4.1Hz,1H),4.61(s,1H),4.52(d,J=10.4Hz,1H),4.22(t,J=6.7Hz,1H),4.16(d,J=10.4Hz,1H),3.61–3.58(m,1H),2.84(d,J=9.8Hz,1H),2.20(dd,J=12.9,6.4Hz,1H),2.10(dt,J=18.4,8.9Hz,2H),1.63(q,J=7.4Hz,2H),1.29(d,J=2.4Hz,9H),1.18(s,4H),1.07(s,4H),0.90(dd,J=8.5,6.1Hz,2H).

[0080] 1H NMR(600MHz,CDCl3)δ7.05(s,1H),6.54(d,J=10.8Hz,1H),6.17(s,1H),5.54(s,1H),5.17(s,1H),4.73(s,1H),4.52(d,J=10.1Hz,1H),4.42(dd,J=12.9,4.6Hz,1H),4.32(d,J=9.7Hz,2H),3.83(dd,J=10.9,6.7Hz,1H),2.98(d,J=9.6Hz,1H),2.25(dd,J=14.3,8.4Hz,1H),2.10–2.02(m,2H),1.90(tt,J=9.0,5.0Hz,1H),1.50(d,J=6.7Hz,1H),1.45–1.39(m,3H),1.35(d,J=25.7Hz,2H),1.24(s,3H),1.20(s,4H),0.96(d,J=8.0Hz,2H),0.88(d,J=7.0Hz,1H),0.84(d,J=5.2Hz,2H).

[0081] 1 H NMR(600MHz,CDCl3)δ8.03(d,J=7.8Hz,2H),7.57(t,J=7.5Hz,1H),7.54(s,1H),7.44(t,J=7.6Hz,2H),6.61(d,J=10.6Hz,1H),6.15(s,1H),5.51(s,1H),5.44(d,J=3.1Hz,2H),4.70(s,1H),4.55(d,J=10.3Hz,1H),4.45(dd,J=13.1,4.4Hz,1H),4.32(d,J=10.2Hz,1H),3.83(dd,J=10.6,6.5Hz,1H),2.94(d,J=9.7Hz,1H),2.16–2.11(m,1H),2.10–2.03(m,2H),1.69–1.58(m,3H),1.51(d,J=6.6Hz,1H),1.43(t,J=13.0Hz,2H),1.32–1.29(m,1H),1.24(s,3H),1.19(s,3H),0.82(td,J=12.9,8.2Hz,1H).

[0082] 1H NMR (600MHz, CDCl3) δ7.96–7.91(m,1H),6.65–6.58(m,1H),6.18(s,1H),5.54(s,1H),4.71(s,1H),4 .60–4.53(m,2H),4.43(q,J=7.1Hz,2H),4.29(d,J=9.7Hz,1H),3.85(dd,J=10.9,6.7Hz,1H),2.99(d ,J=9.7Hz,1H),2.24(dt,J=14.9,9.1Hz,1H),2.11(dd,J=13.1,6.6Hz,2H),1.68–1.62(m,3H),1.54( d,J=6.7Hz,1H),1.49–1.44(m,2H),1.44–1.40(m,3H),1.25(s,4H),1.21(s,3H),0.90–0.81(m,2H).

[0083] Experimental example

[0084] The MTT assay was used to evaluate the in vitro antiproliferative activities of all synthesized C1-triazole oridonin derivatives 1b, 2b, 3b, 4b, 5b, 6b, 1c, 2c, 3c, 4c, and 5c against human non-small cell lung cancer cells A549 and human breast cancer cells MCF-7. The cells and reagents required for the experiments are shown in Table 1.

[0085] Table 1 Cells and reagents required for the experiment

[0086]

[0087]

[0088] The specific operations are as follows:

[0089] ① Cell recovery: Take the frozen cells out of the liquid nitrogen tank and quickly thaw them in a 37°C constant temperature water bath. After the cells in the cryopreserved tube are thawed, use a pipette to transfer the cells in the cryopreserved tube into a centrifuge tube, add 2mL of culture medium, and centrifuge for three minutes (800rpm). Discard the supernatant, add 1mL of culture medium to the centrifuge tube, and use a pipette to gently blow the cells to mix them. Prepare another culture flask, add 4-5mL of culture medium to it, evenly add the blown and mixed cells into the culture flask, and shake gently to mix. Place in an incubator at 37°C and 5% CO2, and change the medium or subculture after 24 hours.

[0090] ②Cell passaging: When the cells in the culture flask grow to about 80-90%, the passaging operation can be carried out. Discard the supernatant, rinse the cells twice with 2mL PBS, add trypsin containing 25% EDTA, and digest the cells for 1-2 minutes. Use a pipette to gently blow the cells so that the cells are no longer attached to the wall, transfer them to a centrifuge tube, and centrifuge for 5 minutes (1000rpm). Discard the supernatant, add 2mL culture medium to the centrifuge tube, and use a pipette to gently blow the cells apart. Prepare two other culture flasks, add 4-5mL culture medium to each, evenly add the blown and mixed cells to the two culture flasks, shake gently to mix, and place in an incubator at 37°C and 5% CO2. Generally, the cells are passaged about twice, and experiments can be carried out when the growth rate reaches the logarithmic period.

[0091] ③Plated: Take cells in a logarithmic growth cycle, discard the supernatant, and rinse the cells twice with 2mL PBS. Add trypsin containing 25% EDTA, digest the cells for 1-2 minutes, gently blow the cells with a pipette so that the cells are no longer attached to the wall, transfer them to a centrifuge tube, and centrifuge for 5 minutes (1000rpm). Discard the supernatant, add 2mL of culture medium to the centrifuge tube, and gently blow the cells evenly with a pipette. Count the cells with a counting plate, configure the appropriate cell concentration, and transfer the cells to a 96-well plate, adding 100μL to each well (5000 cells per well). To avoid edge effects, add PBS solution to the edge wells and set it as a blank control group. Place the 96-well plate in an incubator and culture for 24 hours before adding the drug.

[0092] ④ Drug addition: Dissolve the compound to be tested in DMSO to prepare a 20μM stock solution. Dilute the compound concentration to the experimental concentration using culture medium containing 10% fetal bovine serum. Replace the culture medium with 200μL of fresh culture medium containing the compound at the concentration to be determined, and set up positive control wells, blank control wells, and zero wells. Make 6 replicate wells for each drug or each concentration, and set up a control group and a DMSO group. After drug treatment, place the 96-well plate in a 37°C, 5% CO2 incubator for another 48 hours, add 20μL of CCK-8 solution to each well, continue incubation for 1 hour, and use a microplate reader to detect the absorbance value of each well at a wavelength of 450nm. Calculate the cell inhibition rate according to the following formula.

[0093] Inhibition rate (IR%) = (OD 对照组 -OD 处理组 ) / (OD 对照组 -OD 空白组 )×100%

[0094] The results showed that the CCK-8 method was used to evaluate the in vitro antiproliferative activity of the synthesized C1-triazole oridonin derivatives (1b-6b and 1c-5c) against human non-small cell lung cancer cells A549 and human breast cancer cells MCF-7 at a dosage concentration of 20 μM using oridonin as a positive control (as shown in Table 2). The experimental results are shown in Table 3-1. When the concentration was 20 μM, most of the oridonin triazole derivatives had in vitro antiproliferative activity against A549 and MCF-7 cells. Overall, after the triazole was introduced into oridonin, the in vitro antiproliferative activity against A549 cells was slightly improved compared to the parent oridonin, while the in vitro antiproliferative activity against MCF-7 cells was slightly decreased. The inhibition rate of all compounds against A-549 cells was greater than 60%. The antiproliferative activity of compounds 1b-3b, 5b, 6b, 3c, and 5c was slightly higher than that of oridonin, and the derivative 3c with a cyclopropane substitution on the triazole ring showed the highest inhibitory activity (inhibition rate of 90.3%), which was higher than that of oridonin (inhibition rate of 86.9%). The antiproliferative activity of compounds 4b, 1c, 2c, and 4c was lower than that of oridonin, among which compound 1c had the lowest antiproliferative activity, with an inhibition rate of only 60.5%.

[0095] For MCF-7 cells, the inhibition rates of most compounds were greater than 80%, but all compounds showed slightly lower antiproliferative activity than Rubescensine A. With the exception of compound 2c (inhibition rate of 35.4%), which was much lower than Rubescensine A (inhibition rate of 89.3%), the inhibition rates of the remaining compounds were all greater than 80% but slightly lower than Rubescensine A.

[0096] Table 2 Inhibitory effect of compounds (1b-6b, 1c-5c) on A549 cells and MCF-7 cells

[0097]

[0098] The foregoing description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the form disclosed herein and should not be construed as excluding other embodiments. Rather, the present invention can be used in various other combinations, modifications, and environments and can be modified within the scope of the concept described herein through the above teachings or techniques or knowledge in the relevant field. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention are intended to be protected by the appended claims.

Claims

1. A C1-triazole Rubescensine A derivative as shown in formula (I) or formula (II): Where R is selected from One of the following; Wherein R1 is selected from One of them.

2. The C1-triazole Rubescensine A derivative according to claim 1, characterized in that: The synthesis steps of C1-triazole Rubescensine A derivative I include: Dissolving C1-azidorubescensine A in an organic solvent, adding a catalyst, a ligand, an organic base and an acetylenic compound, heating for reaction, and extracting to obtain a C1-triazolerubescensine A derivative I; The structural formula of the C1-azidorubescensine A c is:

3. The synthesis step according to claim 2, characterized in that: The organic solvent is toluene, benzene, tetrahydrofuran or acetonitrile; The catalyst is CuI, CuBr or CuCl; The ligand is triphenylphosphine, bipyridine, 1,10-phenanthroline or a derivative thereof; The organic base is N,N-diisopropylethylamine, triethylamine, DBU or DIPEA; The heating reaction temperature is 80-85°C; The mass ratio of the C1-triazole Rubescensine A c to the alkyne is 1:10-12.

4. The C1-triazole Rubescensine A derivative according to claim 1, characterized in that: The deketone reaction steps of the C1-triazole oridonin derivative I include: The C1-triazole Rubescensine A derivative I is dissolved in an organic solvent, an acid catalyst is added, and the reaction is carried out to obtain the C1-triazole Rubescensine A derivative II.

5. The deketone reaction according to claim 5, wherein: The organic solvent is tetrahydrofuran, dichloromethane, ether or ethyl acetate; The acid catalyst is concentrated hydrochloric acid, sulfuric acid, trifluoroacetic acid, p-toluenesulfonic acid or aluminum chloride; The reaction temperature is 15-25°C.

6. The C1-triazole Rubescensine A derivative and its synthesis method and deketoneization method according to claims 1-5, characterized in that: The synthesis method of C1-triazole Rubescensine A, The following steps are involved: S1: dissolving oridonin in an organic solvent, adding an acid catalyst, and reacting to obtain oridonin with a protecting group; dissolving the oridonin with a protecting group in an inert organic solvent, adding an organic base and a sulfonylating agent, and reacting to obtain oridonin A a substituted with a 1-methoxysulfonyl group and a protecting group; S2: dissolving the 1-methylsulfonyl-substituted protected Rubescensine A a in a polar aprotic solvent, adding an inorganic base and a halide salt to react to obtain a C1-halogenated Rubescensine A analogue b; S3: dissolving the C1-halogenated oridonin analogue b in an organic solvent, adding an azidation reagent and a catalyst to react, and obtaining the C1-triazole oridonin c.

7. The synthesis method according to claim 6, characterized in that: The organic solvent in step S1 is 2,2-dimethoxypropane; the acid catalyst in step S1 is p-toluenesulfonic acid, sulfuric acid, trifluoroacetic acid, camphorsulfonic acid or Lewis acid; The inert organic solvent is dichloromethane, tetrahydrofuran, ether or ethyl acetate; The sulfonylating agent is methanesulfonyl chloride, p-toluenesulfonyl chloride or trifluoromethanesulfonic anhydride.

8. The synthesis method according to claim 6, wherein: The polar aprotic solvent in step S2 is N,N-dimethylformamide, dimethyl sulfoxide, N-methylpyrrolidone or hexamethylphosphoramide; The inorganic base is lithium carbonate, sodium carbonate, potassium carbonate or cesium carbonate; The halide salt is lithium bromide, sodium bromide, potassium bromide, lithium chloride or sodium iodide.

9. The synthesis method according to claim 6, characterized in that: The organic solvent in step S3 is acetonitrile, N,N-dimethylformamide, tetrahydrofuran or dimethyl sulfoxide; The azidation reagent is trimethylsilyl azide, sodium azide or lithium azide; The catalyst is tetrabutylammonium fluoride, cesium fluoride or potassium fluoride.

10. The use of the C1-triazole Rubescensine A derivative according to claim 1, characterized in that: Used to prepare anticancer drugs.

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