Ring-opening selenine-like organic selenium compound as well as preparation method and application thereof

Abstract

The invention discloses a ring-opening selenine-like organic selenium compound as well as a preparation method and application thereof, and relates to the field of medicinal chemistry. The 11 ring-opening selenine-like organic selenium compounds with novel structural characteristics are successfully synthesized, the synthesis method is simple and convenient, the reaction condition is mild, the yield is high, and the method is suitable for industrial production. Experiments show that the compounds have the following advantages: 1) the toxicity is low, the safety is good, and the effective action concentration is far lower than a toxicity threshold value; 2) the anti-tumor activity is remarkable, an excellent inhibition effect is shown on esophageal cancer, lung cancer, liver cancer, colon cancer and breast cancer, the inhibition rates of the compound 11 on KYS30 and A549 cells at the concentration of 10 mu M are 75% and 85% or above respectively, and the IC50 value is remarkably lower than that of a control drug fumagillin; and 3) in a zebra fish model, the subintestinal blood vessel area and the number of internode blood vessels can be obviously reduced, which indicates that the compound has an anti-angiogenesis effect. The compound disclosed by the invention has important application value in the aspect of preparing anti-tumor and anti-angiogenesis medicines.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a ring-opened selenoline-like organic selenium compound, a preparation method and an application thereof. Background Art

[0002] Selenium is one of the essential trace elements for the human body, and selenium-containing substances are closely related to cancer. Scientific research data from pharmacy, preventive medicine, nutrition, experimental animal science, pharmacology, and clinical medicine have strongly demonstrated the ability of organic selenium compounds to inhibit cancer cell proliferation, prevent the occurrence, development, and deterioration of cancer, assist cancer patients in improving their quality of life and facilitating recovery, and directly kill cancer cells. As a class of molecules with anti-cancer potential, organic selenium compounds have anti-tumor mechanisms characterized by multi-target, multi-enzymatic, and broad-spectrum properties. Therefore, organic selenium compounds have become a hot topic in the development of anti-cancer drugs. Synthetic organic selenium compounds have stronger anti-tumor activity and lower toxicity than inorganic and natural selenium compounds, and are expected to become an important means of comprehensive cancer treatment.

[0003] Selenoid compounds, exemplified by ebselen, possess glutathione peroxidase (GPx)-like activity and antioxidant properties. They also exert anti-tumor effects by specifically inhibiting thioredoxin reductase (TrxR) activity, blocking downstream signaling and anti-apoptotic pathways in the thioredoxin system, and selectively inducing apoptosis in tumor cells. The key anti-tumor pharmacophore of selenoid compounds was once believed to be the selenoid ring, a selenium-containing heterocyclic structure of benzisoselenazolone, and subsequent research has largely focused on this ring. For example, in 2009, two ebselen derivatives designed by Zhang Hua et al. demonstrated significant inhibition of liver cancer cell growth. In 2010, ethaselen, developed by Zeng Huihui's team, was used to treat non-small cell lung cancer with high TrxR expression. Recently, selenoid compounds have been found to be potential slow-binding inhibitors of methionyl aminopeptidase 2 (MetAP2), with their anti-cancer mechanism potentially involving anti-angiogenesis. In the process of continuous follow-up research on the anti-tumor activity of ebselen and its derivatives, the inventor team found that the selenoid ring (benzisoselenazolone five-membered ring) is susceptible to nucleophilic attack by ketone compounds, and the selenium-nitrogen chemical bond (Se-N) breaks and opens, and Se-C bond coupling occurs with the sp3 carbon atom at the α position of the ketone carbonyl group to form a new organic selenium compound with an open-ring selenoid structure (selenoether amide mother nucleus). After a series of studies (ZL202110915433.X; ZL201910935132.6; ZL201910105153.5), it was confirmed that the anti-tumor activity of the open-ring selenoid is better than that of the corresponding unopened selenoid mother drug, breaking the original conclusion that the key anti-tumor pharmacophoric group is the selenoid ring, and proposing that the open-ring selenoid is a better form to exert good anti-tumor activity.

[0004] The previous research is based on the open ring selenol original structure, the chemical structure is focused on the research of o-selenyl benzamide derivatives, and the structure-activity relationship of the anticancer activity of different ketones and different amines as substrates is optimized. As a continuation of this series of research, we continue to synthesize and expand the compounds of this structure to the meta and para substituted organic selenium compounds similar to the open ring selenol structure, and study their application value in the field of anti-tumor and other medical fields. SUMMARY

[0005] For the research target of high-activity anti-cancer open ring selenol compounds, the purpose of the present application is to provide a non-classical open ring selenol-like organic selenium compound, including o-, m-, and p-three open ring selenol analogs. The second purpose is to provide a preparation method of the open ring selenol-like organic selenium compound; the third purpose is to provide the application of the open ring selenol-like organic selenium compound in preparing an antitumor drug.

[0006] In order to achieve the above purposes, the technical scheme adopted by the present application is:

[0007] An open ring selenol-like organic selenium compound, the structural formula of which is:

[0008]

[0009] Among them, -HN-R2-NH- is a divalent group formed by removing two hydrogen atoms from diamine H2N-R2-NH2, which is a divalent bridging structure for connecting chloroselenyl benzoyl chloride; H2N-R2-NH2 is one of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, ethylenediamine, diethyltoluenediamine, pyridine-3,5-diamine, and 1,2-cyclohexanediamine; -Se- in -Se-R1- is a H on a benzene ring replaced by -Se-, and the substitution site is the ortho, para or meta position of the amide bond; -R1- is a side chain formed by replacing the alpha-H of a ketone carbonyl in R1 with -Se-; and R1 is one of acetone, 2-butanone, phenylethanone, cyclobutanone, 2-indanone, and 5-hexen-2-one.

[0010] Further, the structural formula of the compound is:

[0011]

[0012] A preparation method of an open ring selenol-like organic selenium compound, the preparation reaction formula is as follows:

[0013]

[0014] The component a is one of o-chloroselenyl benzoyl chloride, m-chloroselenyl benzoyl chloride and p-chloroselenyl benzoyl chloride; the component b is one of acetone, 2-butanone, phenylacetone, cyclobutanone, 2-indanone and 5-hexen-2-one; and the component c is one of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, ethylenediamine, diethyltoluenediamine, pyridine-3,5-diamine and 1,2-cyclohexanediamine.

[0015] Further, the preparation method comprises the following steps: dissolving the component a in a solvent, adding the component b, stirring and reacting at 15-45 DEG C for 2-36 hours, adding the component c, stirring at 15-45 DEG C for 0.5-1.5 hours, adding a small amount of ethyl acetate to dilute the solution, standing to precipitate the solid, filtering, washing and drying; and the crude product is separated and purified by column chromatography to obtain the open-ring selenium compound.

[0016] Further, the molar ratio of the component a, the component b and the component c is 1:1.5:0.5.

[0017] Further, the solvent is one of tetrahydrofuran, dioxane, toluene, acetone and dichloromethane.

[0018] The open-ring selenium compound is used for preparing an anti-tumor drug.

[0019] Further, the anti-tumor drug is a drug for treating esophageal cancer, liver cancer, breast cancer, lung cancer and colon cancer.

[0020] The open-ring selenium compound is used for preparing an anti-angiogenesis drug.

[0021] According to the above technical solution, the application has the following advantages:

[0022] The application breaks through the limitation of the classical open-ring selenium compound in structure design, successfully synthesizes 11 kinds of representative open-ring selenium compounds with novel structural characteristics by highlighting the selenium functional group in the ortho, meta and para substitution sites and using diamine as a linker, and has the advantages of simple synthesis method, mild reaction condition, high yield, good industrial production prospect, excellent biological activity and safety, and far lower effective concentration than the toxicity threshold.

[0023] In terms of biological activity, the selenolane-like organic selenium compound provided by the present application exhibits significant anti-tumor growth effect, and the compound exhibits excellent inhibitory activity on various tumor cells such as esophageal cancer, lung cancer, liver cancer, colon cancer and breast cancer, and the inhibitory effect is significantly better than that of the control drug fumagillin; through in-vitro anti-tumor experiments, it is found that the compound has significant inhibitory effect on human esophageal squamous cell carcinoma cells (KYS30) and non-small cell lung cancer cells (A549), and in particular, the inhibitory rate of the compound 11 on the human esophageal squamous cell carcinoma cells (KYS30) is more than 75% at a concentration of 10 μM, and the inhibitory rate on the non-small cell lung cancer cells (A549) is more than 85%. Further, the inhibitory effect of the test compound 11 on 11A549 (lung cancer cell line), KYSE30 (esophageal cancer cell line), HepG2 (liver cancer cell), HCT116 (colon cancer cell), MDA-MB-231 (breast cancer cell) is tested, and the results show that the inhibitory effect of the compound 11 is significantly better than that of the fumagillin group, and the IC 50 value is much lower than that of the fumagillin group, indicating that the anti-tumor effect is better.

[0024] Meanwhile, the compound exhibits obvious anti-angiogenic effect in a zebrafish model, and the present application further uses zebrafish as an experimental model to study the anti-angiogenic effect of the compound 11, and the experimental results show that the compound 11 significantly reduces the area of the subintestinal vessel of the zebrafish and the number of complete intersegmental vessels, indicating that the compound has anti-angiogenic effect and good safety, and has significant application value in the field of anti-angiogenic medicine. The above research results further indicate that the selenolane-like organic selenium compound synthesized in the present application has significant anticancer activity on esophageal cancer, lung cancer, liver cancer, colon cancer and breast cancer. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 Fig. 1 is a typical diagram of the area of the subintestinal vessel of zebrafish after sample treatment in the embodiment of the present application;

[0026] Figure 2 Fig. 2 is a columnar diagram of the area of the subintestinal vessel of zebrafish after sample treatment in the embodiment of the present application;

[0027] Figure 3 Fig. 3 is a typical diagram of the number of complete intersegmental vessels of zebrafish after sample treatment in the embodiment of the present application;

[0028] Figure 4 Fig. 4 is a columnar diagram of the number of complete intersegmental vessels of zebrafish after sample treatment in the embodiment of the present application. DETAILED DESCRIPTION

[0029] The present application will be further described in detail below in combination with the drawings and specific embodiments.

[0030] The preparation reaction formula of the selenolane-like organic selenium compounds 1-11 is as follows:

[0031]

[0032] The preparation method of the class ring-opening selenazoline organic selenium compounds 1-11 comprises the following steps:

[0033] The component a is dissolved in a solvent, after the component b is added, the reaction is stirred at 15-45℃ for 2-36h, the component c is added, after the stirring at 15-45℃ for 0.5-1.5h, a small amount of ethyl acetate is added to dilute the solution, after the solid is precipitated, the filtration, washing and drying are conducted, and the column chromatography is adopted to separate and purify the crude product, so as to obtain the class ring-opening selenazoline organic selenium compound. The component a is one of o-chloroselenyl benzoyl chloride, m-chloroselenyl benzoyl chloride and p-chloroselenyl benzoyl chloride; the component b is one of acetone, 2-butanone, phenylethanone, cyclobutanone, 2-indanone and 5-hexen-2-one; and the component c is one of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, ethylenediamine, diethyltoluenediamine, pyridine-3,5-diamine and 1,2-cyclohexanediamine.

[0034] Eleven representative compounds of the class ring-opening selenazoline organic selenium compound are prepared in the examples, and the structures are as follows:

[0035]

[0036] Example 1

[0037] 0.25g (1mmol) of m-chloroselenyl benzoyl chloride is dissolved in 5mL of dichloromethane, 1.5mmol of cyclobutanone is added, after the stirring at 25℃ for 24h, 0.5mmol of diethyltoluenediamine is added, after the stirring at 25℃ for 40min, 1ml of ethyl acetate is added to dilute the solution, after the solid is precipitated, the filtration, washing and drying are conducted, and the column chromatography (dichloromethane:methanol=10:1) is adopted to separate and purify the crude product, so as to obtain the pure compound 1.

[0038] Compound 1: brown solid, melting point 207-208℃, yield 63%, R f =0.65 (dichloromethane:methanol (V / V)=10:1). The structure is analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results are as follows: 1H NMR (400 MHz, DMSO-d6) δ 9.96 (d, J = 6.7, 6.0 Hz, 2H), 8.31 ~ 8.19 (m, 2H), 7.94 ~ 7.82 (m, 4H), 7.56 ~ 7.45 (m, 2H), 7.12 ~ 7.06 (m, 2H), 6.87 ~ 6.81 (m, 1H), 5.75 (s, 2H), 2.75 ~ 2.64 (m, 4H), 2.55 ~ 2.34 (m, 4H), 2.18 (s, 2H), 2.10 (d, J = 2.5 Hz, 2H), 1.25 ~ 1.15 (m, 3H), 1.15 ~ 0.95 (m, 6H). 13 C NMR (101 MHz, DMSO) δ 206.44, 165.93, 165.54, 135.77, 134.28, 134.18, 133.50, 132.08, 130.95, 130.42, 130.18, 129.30, 129.20, 128.84, 127.38, 51.72, 45.87, 24.73, 23.82, 23.73, 20.50, 14.78, 14.76. HRMS (ESI): m / z 683.0923 [M+H] + (calcd. for C 33 H 35 N2O4S e2 + ,683.0949).

[0039] The cell growth inhibition rate of compound 1 on human esophageal squamous cell carcinoma cells (KYS30) was 15.81 ± 1.88% at a concentration of 10 μM, and the cell growth inhibition rate on non-small cell lung cancer cells (A549) was 11.74 ± 0.20%.

[0040] Example 2

[0041] 0.25 g (1 mmol) of m-chloroselenyl benzoyl chloride was dissolved in 5 mL of dichloromethane, 1.5 mmol of 2-indanone was added, and after stirring at 35°C for 36 h, 0.5 mmol of diethyltoluene diamine was added, and after stirring at 35°C for 40 min, 1 mL of ethyl acetate was added to dilute the solution, and after the solid was allowed to precipitate, it was filtered, washed, and dried, and the crude product was separated and purified by column chromatography (dichloromethane:methanol = 1:1) to obtain pure compound 2.

[0042] Compound 2: yellow solid, melting point 179-180°C, yield 52%, R f = 0.72 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1H NMR (400 MHz, DMSO-d6) δ 9.29 (s, 1H), 9.16 (s, 1H), 8.00 ~ 7.96 (m, 2H), 7.87 ~ 7.82 (m, 2H), 7.71 ~ 7.66 (m, 2H), 7.64 ~ 7.56 (m, 2H), 7.47 ~ 7.41 (m, 2H), 7.41 ~ 7.35 (m, 2H), 7.23 ~ 7.14 (m, 4H), 7.05 (t, J = 1.0 Hz, 1H), 5.45 (s, 2H), 3.53 ~ 3.46 (m, 4H), 2.78 ~ 2.62 (m, 4H), 2.15 (s, 3H), 1.23 (t, J = 7.1 Hz, 3H), 1.16 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, DMSO) δ 211.67, 211.61, 167.08, 165.63, 141.46, 141.02, 140.99, 139.78, 139.55, 139.53, 138.48, 138.47, 137.64, 135.64, 135.60, 134.54, 134.50, 133.63, 133.59, 129.54, 129.53, 127.92, 127.86, 127.60, 127.57, 127.36, 127.35, 126.61, 125.92, 125.91, 124.96, 123.89, 123.87, 116.21, 47.31, 47.25, 43.08, 42.99, 25.18, 20.87, 14.74, 14.60, 13.55. HRMS (ESI): m / z 829.1042 [M+Na] + (calcd. for C 43 H 38 N2O4Se2Na + ,823.1062)。

[0043] Compound 2 showed 14.94 ± 0.98% of cell growth inhibition on human esophageal squamous cell carcinoma cells (KYS30) and 13.97 ± 1.26% of cell growth inhibition on non-small cell lung cancer cells (A549) at a concentration of 10 μM.

[0044] Example Three

[0045] 0.25 g (1 mmol) of m-chloroselenobenzoyl chloride was dissolved in 5 mL of dichloromethane, and 1.5 mmol of cyclobutanone was added. After stirring at 15°C for 24 h, 0.5 mmol of 1,2-cyclohexanediamine was added. After further stirring at 25°C for 90 min, 1 ml of ethyl acetate was added to dilute the solution. After standing, a solid precipitated, which was filtered, washed, and dried. The crude product was separated and purified by column chromatography (dichloromethane: methanol = 10:1) to obtain pure compound 3.

[0046] Compound 3: white solid, melting point 185-186°C, yield 69%, R f =0.35 (dichloromethane:methanol (V / V)=10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results are as follows: 1 H NMR (400MHz, DMSO-d6) δ7.83~7.80(m,2H),7.79~7.75(m,2H),7.67~7.63(m,2H),7.53(d,J=7.5Hz,2H),7.38~7.33(m,2H),4.41(t,J=4 .8Hz,2H),3.52(dt,J=6.3,2.9Hz,2H),2.10~1.96(m,2H),1.84~1.72(m,4H),1.73~1.61(m,4H),1.48~1.34(m,4H),1.25~1.10(m,2H). 13 C NMR (101MHz, DMSO) δ208.78,208.73,166.83,166.81,137.74,137.73,134.81,134.77,134.73,134.69,132.16,132.13,129.55 ,129.53,126.56,126.54,56.68,56.63,53.49,53.47,35.86,35.84,30.24,30.22,23.47,23.46,23.44,23.41.HRMS(ESI):m / z 618.0519[M] + (calcd.for C 28 H 30 N2O4Se2,618.0536).

[0047] At a concentration of 10 μM, compound 3 had a cell growth inhibition rate of 11.58±0.74% on human esophageal squamous cell carcinoma cells (KYS30) and a cell growth inhibition rate of 23.80±3.36% on non-small cell lung cancer cells (A549).

[0048] Example 4

[0049] Compound 4: white solid, melting point 244-245℃, yield 72%, Rf=0.71 (dichloromethane:methanol (V / V)=10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR) as follows:

[0050] Compound 4: white solid, melting point 244-245℃, yield 72%, Rf=0.71 (dichloromethane:methanol (V / V)=10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR) as follows: f =0.71 (dichloromethane:methanol (V / V)=10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR) as follows: 1 HNMR (400 MHz, DMSO-d6) δ 9.07 (s, 1H), 8.95 (s, 1H), 8.02-7.97 (m, 2H), 7.85-7.80 (m, 2H), 7.55-7.48 (m, 2H), 7.33-7.26 (m, 2H), 7.02 (t, J=1.0 Hz, 1H), 4.10 (q, J=6.8 Hz, 2H), 2.85-2.70 (m, 4H), 2.23 (s, 9H), 1.13 (t, 6H), 1.06 (q, J=7.5 Hz, 6H). 13 C NMR (101 MHz, DMSO) δ 204.25, 204.20, 167.50, 167.32, 141.46, 138.86, 136.94, 136.82, 136.77, 136.08, 135.77, 133.74, 133.73, 131.84, 131.82, 130.61, 130.60, 127.42, 127.39, 126.48, 124.92, 116.25, 43.00, 42.94, 28.59, 28.58, 25.18, 20.87, 17.46, 17.43, 14.74, 14.60, 13.55. HRMS (ESI): m / z 709.1048 [M+Na] + (calcd. for C 33 H 38 N2O4Se2Na + ,709.1062).

[0051] Compound 4 showed 66.79±0.91% of cell growth inhibition rate on human esophageal squamous cell carcinoma cells (KYS30) and 63.03±1.13% of cell growth inhibition rate on non-small cell lung cancer cells (A549) at a concentration of 10 μM.

[0052] Example Five

[0053] To 0.25 g (1 mmol) of o-chloroselenyl benzoyl chloride was dissolved in 5 mL of dichloromethane, 1.5 mmol of acetone was added, and after stirring at 25°C for 2 h, 0.5 mmol of diethyltoluenediamine was added, and stirring was continued at 15°C for 40 min, and then 1 mL of ethyl acetate was added to dilute the solution, and after the solid was allowed to precipitate, it was filtered, washed, and dried, and the crude product was separated and purified by column chromatography (dichloromethane:methanol = 10:1) to obtain pure compound 5.

[0054] Compound 5: white solid, melting point 208-209°C, yield 76%, Rf= 0.67 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: f = 0.67 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 2H), 7.85-7.76 (m, 2H), 7.65-7.57 (m, 2H), 7.48-7.43 (m, 2H), 7.41-7.33 (m, 2H), 7.13-7.06 (m, 1H), 3.82 (s, 4H), 2.59 (d, J = 10.3 Hz, 4H), 2.24 (q, J = 2.5 Hz, 9H), 1.22-1.14 (m, 3H), 1.11-1.04 (m, 3H). 13 CNMR (101 MHz, DMSO) δ 204.88, 167.93, 141.30, 135.98, 135.83, 133.17, 132.68, 132.49, 131.44, 130.16, 128.47, 128.40, 128.06, 126.10, 35.66, 28.55, 28.49, 24.94, 18.61, 15.21. HRMS (ESI): m / z 659.0931 [M+H] + (calcd. for C 31 H 35 N2O4Se2 + , 659.0949).

[0055] Compound 5 had a cell growth inhibition rate of 26.95 ± 1.16% on human esophageal squamous cell carcinoma cells (KYS30) and a cell growth inhibition rate of 38.32 ± 2.63% on non-small cell lung cancer cells (A549) at a concentration of 10 μM.

[0056] Example Six

[0057] Compound 6: white solid, melting point 164-165 °C, yield 49%, Rf= 0.54 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows:

[0058] Compound 6: white solid, melting point 164-165 °C, yield 49%, Rf= 0.54 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: f = 0.54 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 7.98-7.90 (m, 4H), 7.55-7.48 (m, 2H), 7.32 (d, J = 7.5 Hz, 2H), 7.33-7.26 (m, 2H), 4.21-4.14 (m, 2H), 3.87 (dt, J = 7.5, 4.4 Hz, 2H), 2.93 (dd, J = 5.4, 1.2 Hz, 4H), 2.24 (d, J = 1.4 Hz, 6H). 2.11-2.03 (m, 2H), 1.85-1.69 (m, 2H), 1.71-1.61 (m, 2H), 1.52-1.32 (m, 2H), 1.25-1.08 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 202.66, 202.60, 167.23, 167.22, 134.49, 134.47, 134.46, 134.44, 133.57, 133.53, 132.34, 132.31, 127.74, 127.73, 127.71, 127.70, 80.56, 80.55, 70.13, 70.12, 54.35, 54.29, 53.49, 53.48, 30.24, 30.22, 28.62, 28.60, 23.47, 23.44, 20.37, 20.34. HRMS (ESI): m / z 673.1557 [M-H] - (calcd. for C 32 H 37 N2O4Se2 - ,673.1562).

[0059] Compound 6 had a cell growth inhibition rate of 26.72 ± 0.45% on human esophageal squamous cell carcinoma cells (KYS30) and a cell growth inhibition rate of 41.49 ± 4.85% on non-small cell lung cancer cells (A549) at a concentration of 10 μM.

[0060] Example Seven

[0061] To a solution of 0.25 g (1 mmol) of p-chloroselenyl benzoyl chloride in 5 mL of dichloromethane, 1.5 mmol of 2-indanone was added, and the mixture was stirred at 45 °C for 36 h. Then, 0.5 mmol of ethylenediamine was added, and the mixture was stirred at 20 °C for 30 min. After dilution with 1 mL of ethyl acetate, the solid was separated by filtration, washed, and dried. The crude product was separated and purified by column chromatography (dichloromethane:methanol = 1:1) to obtain pure compound 7.

[0062] Compound 7: gray solid, melting point 223-224 °C, yield 64%, Rf= 0.32 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: f = 0.32 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 7.85-7.79 (m, 4H), 7.72 (t, J = 5.2 Hz, 2H), 7.63-7.56 (m, 2H), 7.58-7.52 (m, 4H), 7.41-7.35 (m, 2H), 7.23-7.14 (m, 4H), 5.47 (s, 2H), 3.52 (d, J = 5.2 Hz, 4H), 3.51 (d, J = 0.9 Hz, 2H), 3.48 (d, J = 1.1 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 211.68, 211.62, 168.35, 168.30, 141.13, 141.10, 139.56, 139.54, 135.00, 134.99, 134.97, 134.96, 133.18, 133.15, 132.62, 132.58, 128.14, 128.12, 128.11, 128.10, 127.93, 127.87, 127.61, 127.59, 125.93, 125.92, 123.90, 123.89, 57.29, 57.23, 53.04, 53.00, 49.28, 49.23. HRMS (ESI): m / z 688.0398 [M] + (calcd for C 34 H 28 N2O4Se2 + , 688.0380).

[0063] The cell growth inhibition rate of compound 7 on human esophageal squamous cell carcinoma cells (KYS30) was 33.00±1.03% at a concentration of 10 μM, and the cell growth inhibition rate on non-small cell lung cancer cells (A549) was 33.41±3.39%.

[0064] Example Eight

[0065] 0.25 g (1 mmol) of p-chloroselenyl benzoyl chloride was dissolved in 5 mL of dichloromethane, 1.5 mmol of 2-indanone was added, and after stirring at 45°C for 36 h, 0.5 mmol of diethyltoluene diamine was added, and after stirring at 35°C for 40 min, 1 mL of ethyl acetate was added to dilute the solution, and after the solid was allowed to precipitate, it was filtered, washed, and dried, and the crude product was separated and purified by column chromatography (dichloromethane:methanol = 1:1) to obtain pure compound 8.

[0066] Compound 8: yellow solid, melting point 179-180°C, yield 54%, Rf f = 0.65 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 9.98 (s, 1H), 9.87 (s, 1H), 8.06-7.86 (m, 6H), 7.86-7.69 (m, 4H), 7.42-7.21 (m, 4H), 7.12 (dd, J = 7.7, 3.1 Hz, 2H), 6.98 (d, J = 8.9 Hz, 1H), 5.76 (s, 2H), 4.02-3.98 (m, 4H), 2.84-2.73 (m, 4H), 2.27 (s, 3H), 1.23-1.09 (m, 6H). 13 C NMR (101 MHz, DMSO) δ 212.58, 212.52, 167.64, 167.30, 142.37, 142.03, 141.99, 140.67, 140.46, 140.44, 138.01, 135.83, 135.82, 135.80, 135.79, 134.09, 134.06, 133.84, 133.70, 129.30, 129.29, 129.28, 129.26, 128.83, 128.77, 128.51, 128.48, 127.62, 126.83, 126.82, 125.87, 124.80, 124.78, 117.12, 47.31, 47.25, 43.08, 42.99, 26.09, 21.78, 15.65, 15.51, 14.46. HRMS (ESI): m / z 829.1049 [M+Na] +(calcd. for C 43 H 38 N2O4Se2Na + ,829.1062)。

[0067] Compound 8 showed 43.06 ± 3.12% of cell growth inhibition on human esophageal squamous cell carcinoma (KYS30) and 36.08 ± 6.10% of cell growth inhibition on non-small cell lung cancer (A549) at a concentration of 10 μM.

[0068] Example Nine

[0069] Dissolve 0.25 g (1 mmol) of p-chloroselenyl benzoyl chloride in 5 mL of dichloromethane, add 1.5 mmol of 2-indanone, and stir at 25°C for 36 h. Add 0.5 mmol of pyridine-3,5-diamine, and continue stirring at 20°C for 60 min. Add 1 mL of ethyl acetate to dilute the solution, and allow the solid to precipitate. Filter, wash, and dry the solid. Purify the crude product by column chromatography (dichloromethane:methanol = 1:1) to obtain pure compound 9.

[0070] Compound 9: brown solid, melting point 189-190°C, yield 43%, R f = 0.55 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance spectroscopy (NMR) as follows: 1 H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 1.8 Hz, 2H), 8.10 (t, J = 1.8 Hz, 1H), 7.90-7.83 (m, 4H), 7.63-7.55 (m, 6H), 7.41-7.35 (m, 2H), 7.23-7.14 (m, 4H), 5.47 (s, 2H), 3.49 (dd, J = 15.1, 1.0 Hz, 4H). 13 C NMR (101 MHz, DMSO) δ 211.68, 211.63, 166.55, 166.49, 141.21, 141.18, 141.14, 141.10, 139.57, 139.55, 137.52, 137.49, 134.94, 134.93, 134.91, 134.90, 133.20, 133.17, 132.98, 132.94, 128.41, 128.40, 128.38, 128.37, 127.94, 127.88, 127.62, 127.59, 125.94, 125.92, 123.91, 123.89, 115.04, 51.93, 50.02. HRMS (ESI): m / z 737.0353 [M] +(calcd. for C 37 H 27 N3O4Se2 + ,737.0332)。

[0071] Compound 9 showed 44.24±2.66% of cell growth inhibition rate on human esophageal squamous cell carcinoma (KYS30) and 47.38±1.37% of cell growth inhibition rate on non-small cell lung cancer (A549) at 10 μM concentration.

[0072] Example Ten

[0073] Dissolve 0.25 g (1 mmol) of p-chloroselenyl benzoyl chloride in 5 mL of dichloromethane, add 1.5 mmol of phenylacetone, stir at 45°C for 18 h, add 0.5 mmol of 1,2-cyclohexanediamine, continue stirring at room temperature for 90 min, add 1 mL of ethyl acetate dilution solution, allow solid to precipitate, filter, wash, and dry, separate and purify the crude product by column chromatography (dichloromethane:methanol = 10:1) to obtain pure compound 10.

[0074] Compound 10: white solid, melting point 255-256°C, yield 68%, R f = 0.48 (dichloromethane:methanol (V / V) = 10:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 7.92-7.84 (m, 4H), 7.84-7.78 (m, 4H), 7.55-7.49 (m, 4H), 7.52-7.44 (m, 6H), 7.33 (d, J = 7.5 Hz, 2H), 4.62 (d, J = 4.4 Hz, 4H), 3.83 (dt, J = 7.5, 4.4 Hz, 2H), 1.86 (dddd, J = 13.2, 8.8, 6.1, 4.5 Hz, 2H), 1.68 (dddd, J = 12.9, 8.8, 6.1, 4.4 Hz, 2H), 1.58 (ddd, J = 13.0, 8.9, 6.1 Hz, 2H), 1.43 (ddd, J = 13.0, 9.1, 6.1 Hz, 2H). 13C NMR (101 MHz, DMSO) d 188.77, 188.71, 167.23, 167.22, 136.33, 136.30, 133.59, 133.55, 133.19, 133.16, 132.74, 132.72, 132.71, 132.69, 132.08, 132.05, 128.90, 128.87, 128.84, 128.82, 128.81, 128.79, 128.77, 127.32, 127.31, 127.30, 127.28, 53.49, 53.48, 30.67, 30.62, 30.24, 30.22, 23.47, 23.44. HRMS (ESI): m / z 741.0730 [M+H] + (calcd. for C 36 H 34 N2O4Se2Na + ,741.0749)。

[0075] Compound 10 showed 41.83 ± 0.62% of cell growth inhibition on human esophageal squamous cell carcinoma (KYS30) and 47.36 ± 4.70% of cell growth inhibition on non-small cell lung cancer (A549) at a concentration of 10 μM.

[0076] Example Eleven

[0077] Dissolve 0.25 g (1 mmol) of m-chloroselenyl benzoyl chloride in 5 mL of dichloromethane, add 1.5 mmol of 2-indanone, stir at 25°C for 36 h, then add 0.5 mmol of 1,2-cyclohexanediamine, continue stirring at 25°C for 90 min, then add 1 mL of ethyl acetate dilution solution, let the solid precipitate, filter, wash, and dry, and separate and purify the crude product by column chromatography (dichloromethane:methanol = 1:1) to obtain pure compound 11.

[0078] Compound 11: white solid, melting point 169-170°C, yield 55%, R f = 0.24 (dichloromethane:methanol (V / V) = 1:1). The structure was analyzed by nuclear magnetic resonance hydrogen spectrum and carbon spectrum (NMR), and the results are as follows: 1HNMR (400 MHz, DMSO-d6) δ 7.98 ~ 7.86 (m, 2H), 7.78 ~ 7.73 (m, 2H), 7.69 ~ 7.63 (m, 2H), 7.61 ~ 7.54 (m, 2H), 7.50 (d, J = 7.5 Hz, 2H), 7.39 ~ 7.30 (m, 4H), 7.21 ~ 7.12 (m, 4H), 5.45 ~ 5.41 (m, 2H), 3.82 (dt, J = 7.5, 4.4 Hz, 2H), 3.47 (dd, J = 15.1, 1.0 Hz, 4H), 1.90 ~ 1.80 (m, 2H), 1.72 ~ 1.62 (m, 2H), 1.62 ~ 1.53 (m, 2H), 1.47 ~ 1.37 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 211.67, 211.61, 166.83, 166.81, 141.02, 140.99, 139.55, 139.53, 138.45, 138.44, 135.22, 135.19, 135.18, 135.15, 133.55, 133.52, 129.57, 129.56, 127.92, 127.86, 127.60, 127.57, 126.60, 126.59, 125.92, 125.91, 123.89, 123.87, 53.49, 53.47, 47.31, 47.25, 43.03, 42.99, 30.24, 30.22, 23.47, 23.44. HRMS (ESI): m / z 765.0747 [M + Na] + (calcd. for C 38 H 34 N2O4Se2Na + ,765.0749).

[0079] Compound 11 showed 75.81 ± 1.88% of cell growth inhibition rate on human esophageal squamous cell carcinoma cells (KYS30) and 86.74 ± 2.00% of cell growth inhibition rate on non-small cell lung cancer cells (A549) at a concentration of 10 μM. In this example, fumagillin was selected as a positive control drug for anti-angiogenesis, and fumagillin showed 43.36 ± 0.45% of cell growth inhibition rate on human esophageal squamous cell carcinoma cells (KYS30) and 46.83 ± 2.26% of cell growth inhibition rate on non-small cell lung cancer cells (A549) at a concentration of 10 μM. As shown in Table 1, compound 11 showed significant inhibitory activity on various cancer cells, and the IC 50 value was much lower than that of fumagillin, indicating that it had a better anti-tumor effect.

[0080] Table 1 Comparison of half inhibitory concentration (IC50) of compound 11 and fumagillin on different tumor cell lines 50 )Comparison

[0081]

[0082] In this embodiment, zebrafish is selected as the experimental model to study the anti-angiogenic effect of compound 11.

[0083] Anti-angiogenic effect evaluation (subintestinal vessels): 6hpf transgenic blood vessel green fluorescent zebrafish (Fli-1) strain was randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Compound 11 was administered in water, and the positive control sorafenib was 0.500 μM in concentration, and a normal control group was set up at the same time, and the volume in each well was 3 mL. After treatment at 28°C for 72 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photography, and the data were collected using NIS-Elements D 3.20 advanced image processing software, and the subintestinal vessel area (pixels) of zebrafish was analyzed, and the statistical analysis results of the index were used to evaluate the anti-angiogenic effect of the sample. The statistical processing results are expressed as mean ± SE. Statistical analysis was performed using SPSS software, and p<0.05 indicates that the difference is statistically significant.

[0084] Anti-angiogenic effect evaluation (intersegmental vessels): 6hpf transgenic blood vessel green fluorescent zebrafish (Fli-1) strain was randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Compound 11 was administered in water, and the positive control sorafenib was 0.500 μM in concentration, and a normal control group was set up at the same time, and the volume in each well was 3 mL. After treatment at 28°C for 48 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photography, and the data were collected using NIS-Elements D 3.20 advanced image processing software, and the number of intersegmental vessels (roots) of zebrafish was analyzed, and the statistical analysis results of the index were used to evaluate the anti-angiogenic effect of the sample. The statistical processing results are expressed as mean ± SE. Statistical analysis was performed using SPSS software, and p<0.05 indicates that the difference is statistically significant.

[0085] The half lethal concentration IC 50 value of zebrafish is 50 μM-100 μM, and the maximum detection concentration (MTC) of zebrafish is 6.25 μM-12.5 μM. As shown in Tables 2, 3 and Figures 1 to 4 , Figure 1 , and Figure 3 The yellow dashed area in the middle is the analysis site, and the experimental results show that compound 11 significantly reduces the subintestinal vessel area and the number of intersegmental vessels of zebrafish, indicating that it has an anti-angiogenic effect and good safety.

[0086] Table 2 Anti-angiogenic effect evaluation experiment results of samples - subintestinal blood vessels (n=10)

[0087]

[0088] ***p<0.001 compared with the normal control group

[0089] Table 3 Anti-angiogenic effect evaluation experiment results of samples - intercalary blood vessels (n=10)

[0090]

[0091] **p<0.01, ***p<0.001 compared with the normal control group

[0092] Related experiments:

[0093] CCK-8 method for detecting cytotoxicity experiment: tumor cells were taken out from liquid nitrogen and quickly placed in a 37°C water bath, the freeze tube was shaken gently to dissolve the freeze solution, and after dissolution, the cells were transferred to a centrifuge tube containing 5 ml of culture medium, and the cells were collected by centrifugation at 1000 rpm for 5 min at room temperature. The supernatant was discarded, and the cells were suspended with complete culture medium containing 10% fetal bovine serum and inoculated into a culture dish, gently mixed by blowing, and cultured at 37°C, 5% CO2 saturated humidity. Cells in the logarithmic growth phase and in good growth condition were inoculated into a cell culture 96-well plate at 4×10 3 cells / well, and cultured overnight in a 37°C, 5% CO2 incubator (100 μL of sterile PBS was added to the wells around the cells). A small amount of sample was placed in an EP tube for sterilization. After sterilization was completed, the sample concentration was set to 20 μM, 10 μM, 5 μM, 2 μM, 1 μM, 0.5 μM, and 0.25 μM. The 96-well plate inoculated with cells the previous day was placed in each concentration of sample for co-culture with the cells. The experimental group had one time point: 24 h, and three groups were set as the experimental group, the control group (with cells and culture medium without drugs), and the blank control group (without cells, with culture medium and without drugs). The plate was placed in the cell culture incubator for 24 h for detection. After 24 h of incubation in the 96-well plate, 10 μL of CCK-8 reagent was added to each well under light-proof conditions, and the plate was placed in the incubator for 3 h. The absorbance value at 450 nm was detected by an enzyme-labeled instrument, and the cell survival rate was calculated. The absorbance values of each group were input into Excel and the relative viability was calculated (% cell relative survival = (experimental group OD value-background OD value) / (control group OD value mean-background OD value) x 100).

[0094] Table 4 IC values of compounds 1-11 on five cancer cell lines 50

[0095] ​

[0096] It should be noted that the above-mentioned embodiments illustrate rather than limit the application, and that those skilled in the art will be able to design many alternative embodiments without departing from the scope of the application. Accordingly, the legal scope of the application is defined only by the claims.

Claims

1. A ring-opened selenophore-like organoselenium compound, characterized in that: Its structural formula is: Wherein, -HN-R2-NH- is a divalent group formed by removing two hydrogen atoms from the diamine H2N-R2-NH2, which serves as a divalent bridging structure connecting chloroselenobenzoyl chloride; the H2N-R2-NH2 is one of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, ethylenediamine, diethyltoluenediamine, pyridine-3,5-diamine, and 1,2-cyclohexanediamine; In the -Se-R1-, -Se is a hydrogen atom on the benzene ring replaced by -Se-, and the substitution site is the ortho, para or meta position of the amide bond; The -R1- is a side chain formed by replacing the α-H of a ketone carbonyl group in R1 by -Se-; The R1 is one of acetone, 2-butanone, acetophenone, cyclobutanone, 2-indanone, and 5-hexen-2-one.

2. The ring-opened selenophene-like organoselenium compound according to claim 1, characterized in that: The structural formula of the compound is:

3. A method for preparing a ring-opened selenophore-like organoselenium compound, characterized in that: The preparation reaction is as follows: Wherein, the component a is one of o-chloroselenobenzoyl chloride, m-chloroselenobenzoyl chloride, and p-chloroselenobenzoyl chloride; The component b is one of acetone, 2-butanone, acetophenone, cyclobutanone, 2-indanone, and 5-hexen-2-one; The component C is one of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, ethylenediamine, diethyltoluenediamine, pyridine-3,5-diamine, and 1,2-cyclohexanediamine.

4. The method for preparing a ring-opened selenophene-like organoselenium compound according to claim 3, wherein: The preparation method comprises the following steps: Component a is dissolved in a solvent, component b is added, and the mixture is stirred at 15°C to 45°C for 2 to 36 hours. Component c is added, and the mixture is stirred at 15°C to 45°C for 0.5 to 1.5 hours. A small amount of ethyl acetate is added to dilute the solution, and the solid is allowed to stand until it precipitates, which is then filtered, washed, and dried. The crude product is separated and purified by column chromatography to obtain the ring-opened selenoline-like organic selenium compound as claimed in claim 1.

5. The method for preparing a ring-opened selenophene-like organoselenium compound according to claim 4, wherein: The molar ratio of component a, component b and component c is 1:1.5:0.

5.

6. The method for preparing a ring-opened selenophene-like organoselenium compound according to claim 4, wherein: The solvent is one of tetrahydrofuran, dioxane, toluene, acetone and dichloromethane.

7. Use of the ring-opened selenol-like organoselenium compound according to claim 1 or claim 2 in the preparation of anti-tumor drugs.

8. The use according to claim 7, characterized in that: The anti-tumor drug is a drug for treating esophageal cancer, liver cancer, breast cancer, lung cancer and colon cancer.

9. Use of the ring-opened selenol-like organoselenium compound according to claim 1 or claim 2 in the preparation of anti-angiogenic drugs.

Images