Organic selenium compound and application thereof in preparation of anti-cancer drugs

Abstract

The invention provides a novel organic selenium compound and a synthesis method thereof, and application of the organic selenium compound in preparation of drugs for preventing and / or treating cancers. According to the compound provided by the invention, a natural active compound is combined with selenium element, so that the anticancer activities of the natural active compound and the selenium element generate a synergistic effect, the safety and the anticancer effect of the compound are improved, and the compound has huge potential in the aspects of preventing and treating cancers.

Description

Technical Field

[0001] The invention belongs to the field of drug synthesis, and specifically relates to a novel organic selenium compound and a synthesis method thereof, as well as application of the compound in preparing drugs for preventing and / or treating cancer. Background Art

[0002] Due to factors such as an aging population, unhealthy lifestyles, and environmental pollution, cancer has become a common and highly prevalent disease worldwide, and the number of cancer patients continues to rise every year. Cancer drug treatments, including chemotherapy and targeted therapy, have made significant progress, but still face many challenges and pain points. Chemotherapy drugs have non-specific toxicity, relatively limited clinical targets, relatively narrow indications for targeted therapy, and are prone to drug resistance, similar to chemotherapy.

[0003] Selenium is one of the essential trace elements in organisms. It has many special functions for human health and is known as the "spark of life" and the "king of anti-cancer". Selenium is the active center of glutathione peroxidase (GSH-Px) and has a strong antioxidant effect. Selenium deficiency in the human body can lead to the occurrence of dozens of diseases. my country has made significant progress in preventing Keshan disease, Kaschin-Beck disease and liver cancer through selenium supplementation. The anti-cancer effect of selenium is mainly achieved through its antioxidant, immunomodulatory, inhibition of cancer cell proliferation and angiogenesis, and promotion of DNA repair. However, most of the selenium used in clinical practice is inorganic selenium, such as sodium selenite and selenium disulfide. However, inorganic selenium has the disadvantages of large toxic side effects and low bioavailability. Currently, a few organic selenium drugs such as ebselen are in the clinical trial stage. There are very few organic selenium drugs that can be used for clinical anti-cancer in the future. Summary of the invention

[0004] In order to overcome the shortcomings in the existing field, the present invention synthesizes a series of organic selenium compounds with the ability to kill cancer cells by reducing, etherifying, amidating and other reactions with selenium and o-methoxybenzylamine. In vitro experiments show that they have good water solubility, anticancer activity and safety.

[0005] In a first aspect, the present invention provides an organic selenium compound having a structure as shown in I-1, I-2, I-3 or I-4: .

[0006] The compound provided by the invention combines selenium with aniline or benzylamine Chinese medicine monomers through chemical bonds to form a new selenium-containing active molecule, so that the two can play a synergistic role.

[0007] In a second aspect, the present invention provides a method for preparing the compound described in the first aspect.

[0008] The organic selenium compound of the present invention is prepared by the following synthetic route: (1) ; (2) ; Among them, R Y represent , , or .

[0009] As a preferred embodiment of the present invention, the step (1) comprises: taking selenium powder, mixing it with distilled water under nitrogen protection, slowly adding sodium borohydride aqueous solution while stirring, and after the system is clarified, adjusting the pH value of 2-chloroacetic acid to 9-10 with saturated sodium carbonate solution, adding it to the above reaction solution, and stirring at room temperature; collecting the reaction solution, adjusting the pH value to 2-3, extracting with ethyl acetate, collecting the organic phase, removing water, concentrating, and recrystallizing to obtain 2,2'-selenodiacetic acid.

[0010] As a preferred embodiment of the present invention, the step (2) comprises: taking dichloromethane, adding N,N'-diisopropylethylamine, 1-hydroxy-7-azobenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 2,2'-selenodiacetic acid and raw material R Y -NH2, stir the reaction at room temperature for 10-15 h, then add water and stir the reaction for another 1-2 h. The resulting reaction solution was separated with a separatory funnel, the aqueous phase was collected, and dichloromethane was added for extraction. The organic phase was collected, dehydrated, and concentrated to obtain a crude product.

[0011] In a third aspect, the present invention provides use of the compound described in the first aspect in the preparation of a drug for preventing and / or treating cancer.

[0012] As a preferred embodiment of the present invention, the cancer is selected from one or more of melanoma, liver cancer, lung cancer, gastric cancer, and cervical cancer.

[0013] As a preferred embodiment of the present invention, the compound has the structure shown in I-1, and the cancer is melanoma or lung cancer.

[0014] As a preferred embodiment of the present invention, the compound has the structure shown in I-2, and the cancer is lung cancer or gastric cancer.

[0015] As a preferred embodiment of the present invention, the compound has the structure shown in I-3, and the cancer is melanoma, lung cancer, gastric cancer or cervical cancer.

[0016] As a preferred embodiment of the present invention, the compound has the structure shown in I-4, and the cancer is melanoma, gastric cancer or cervical cancer.

[0017] As a preferred embodiment of the present invention, the drug exerts anti-cancer efficacy by inhibiting the proliferation of cancer cells, promoting the apoptosis of cancer cells, and inhibiting the synthesis of DNA through one or more mechanisms.

[0018] Compared with the prior art, the present invention combines the natural active compound p-hydroxycinnamic acid with selenium to synthesize a new type of organic selenium drug, filling the gap in the combined use of the anti-cancer active structure of traditional Chinese medicine and selenium, making the anti-cancer activities of the two produce a synergistic effect, and increasing its safety and anti-cancer effect, which has great potential in the prevention and treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 , Compound 1 1 H NMR spectrum (600 MHz, D2O); Figure 2 , ESI-MS spectrum of compound 1; Figure 3 , Compound I-1 1 H NMR spectrum (600 MHz, DMSO- d 6); Figure 4 , ESI-MS spectrum of compound I-1; Figure 5 , Compound I-2 1 H NMR spectrum (600 MHz, DMSO- d 6); Figure 6 , ESI-MS spectrum of compound I-2; Figure 7 , Compound I-3 1 H NMR spectrum (600 MHz, DMSO- d 6); Figure 8 , ESI-MS spectrum of compound I-3; Fig. 9 , Compound I-4 1 H NMR spectrum (600 MHz, DMSO- d 6); Fig.10 , ESI-MS spectrum of compound I-4; Fig.11 , B16 cells, Hela cells, HGC-27 cells and L-O2 cells under different culture conditions. DETAILED DESCRIPTION

[0020] The following examples are provided to further illustrate various aspects of the present invention. These examples are non-limiting and should not be construed as limiting any aspect of the present invention. The scope of protection of the present invention is limited only by the claims. Without departing from the scope of the claims, those skilled in the art may make various modifications and improvements to various aspects of the present invention, and these modifications and improvements also belong to the scope of protection of the present invention.

[0021] In addition, it should be noted that, unless otherwise specified, the various materials and reagents used in the following examples are commonly used materials and reagents in the art and can be obtained through conventional commercial channels; the methods used are conventional methods known to those skilled in the art. Example 1: Synthesis of Compound I-1

[0022] Compound I-1 is 2,2'-selenobis(N-benzylacetamide) with the following structure:

[0023] The following method was used to synthesize compound I-1: (1) Synthesis of 2,2'-selenodiacetic acid (Compound 1): Weigh 1.6 g (0.02 mol) of selenium powder and place it in a 150 ml three-necked flask. Add 10 mL of distilled water and stir under nitrogen. Then slowly drop 1.5 g (0.04 mol) of sodium borohydride dissolved in 10 mL of distilled water. The selenium powder gradually dissolves and the reaction solution becomes clear. Dissolve 3.8 g (0.04 mol) of 2-chloroacetic acid in 20 mL of distilled water, adjust the pH value to 9-10 with saturated sodium carbonate solution, then add it to the above reaction solution and stir at room temperature for 12 h. After the reaction is completed, filter the reaction solution, adjust the pH value of the filtrate to 2-3 with dilute hydrochloric acid, extract with ethyl acetate, dry with anhydrous sodium sulfate, concentrate and crystallize, and then recrystallize with ethyl acetate to obtain 2.58 g of white solid with a yield of 64.9%. 1 H NMR (600MHz, D2O): δ 3.469 (s, 4H) (e.g. Figure 1 ESI-MS ( m / z ): 196.9 [MH] - (like Figure 2 as shown).

[0024] (2) Synthesis of compound I-1: Add 20 ml of dichloromethane (DCM) to a flask, then add 0.38 g (0.003 mol) of N,N'-diisopropylethylamine (DIPEA), 0.205 g (0.0015 mol) of 1-hydroxy-7-azobenzotriazole (HOAT), 0.38 g (0.002 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), 0.197 g (0.001 mol) of compound 1 and 0.214 g (0.002 mol) of benzylamine, and stir at room temperature for 12 h. Add 20 ml of water and stir for 1 h, remove the stirring bar, separate with a separatory funnel, extract with DCM, wash with water, dry, and rotary evaporate DCM to obtain a crude product. The crude product was eluted with a semi-preparative liquid phase using acetonitrile / water (volume ratio 20:70 to 100:0) gradient to obtain the purified product I-10.289 g with a yield of 70.8%. The structural characterization information of compound I-1 is as follows: 1 H NMR (600 MHz, DMSO- d 6) δ 8.518 (t, J = 6.0Hz, 2H), 7.329-7.304 (m, 4H), 7.268-7.227 (m, 6H), 4.271 (d, J = 6.0 Hz, 4H),3.339 (s, 4H) (e.g. Figure 3 ESI-MS( m / z ): 374.9 [MH] - (like Figure 4 as shown). Example 2: Synthesis of Compound I-2

[0025] Compound I-2 is 2,2'-selenobis(N-(2-methoxyphenyl)acetamide) with the following structure:

[0026] By using the method provided in Example 1, only in step (2), 0.214 g (0.002 mol) of benzylamine was replaced with 0.246 g (0.002 mol) of o-anisidine, 0.289 g of product I-2 was synthesized with a yield of 70.8%. The structural characterization information of compound I-2 is as follows: 1 H NMR (600 MHz, DMSO- d 6) δ 9.387 (s, 2H), 7.988 (dd, J=7.8, 1.8 Hz, 2H), 7.076 – 7.017 (m, 4H), 6.89 (td, J = 7.8, 1.8 Hz, 2H), 3.808(s, 6H), 3.3595 (s, 4H) Figure 5 ESI-MS( m / z ): 406.9 [MH] - (like Figure 6 as shown). Example 3: Synthesis of Compound I-3

[0027] Compound I-3 is 2,2'-selenobis(N-(2-methoxybenzyl)acetamide) and has the following structure:

[0028] By using the method provided in Example 1, only 0.214 g (0.002 mol) of benzylamine was replaced with 0.274 g (0.002 mol) of o-methoxybenzylamine in step (2), 0.297 g of product I-3 was synthesized with a yield of 68.1%. The structural characterization information of compound I-3 is as follows: 1 H NMR (600 MHz, DMSO- d 6) δ 8.338 (t, J = 6.0 Hz, 2H),7.237 (td, J = 7.8, 1.8 Hz, 2H), 7.195 (dd, J = 7.8, 1.8 Hz, 2H), 6.971 (dd, J =8.4, 1.2 Hz, 2H), 6.896 (td, J = 7.2, 1.2 Hz, 2H), 4.228 (d, J = 6.0 Hz, 4H),3.789 (s, 6H), 3.333 (s, 4H) (such as Figure 7 ESI-MS( m / z ): 434.9 [MH] - (like Figure 8 as shown). Example 4: Synthesis of Compound I-4

[0029] Compound I-4 is 2,2'-selenobis(N-(3-methoxybenzyl)acetamide) and has the following structure:

[0030] By using the method provided in Example 1, only in step (2), 0.214 g (0.002 mol) of benzylamine was replaced with 0.274 g (0.002 mol) of m-methoxybenzylamine, and 0.306 g of product I-4 was synthesized with a yield of 70.2%. The structural characterization information of compound I-4 is as follows: 1 H NMR (600 MHz, DMSO- d 6) δ 8.504 (t, J = 6.0 Hz, 2H),7.222 (t, J = 7.8 Hz, 2H), 6.836-6.823 (m, 4H), 6.811-6.791 (m, 2H), 4.245 (d, J = 6.0 Hz, 4H), 3.727 (s, 6H), 3.341 (s, 4H) (e.g. Fig. 9 ESI-MS( m / z ): 435.0891[MH] - (like Fig.10 as shown). Experimental example

[0031] This experimental example verifies the anti-tumor activity of compounds I-1 to I-4, using resveratrol and 5-fluorouracil as controls.

[0032] The cell lines involved in this experiment are as follows: (1) mouse melanoma B16 cells, purchased from Beijing Solebow Company, cultured in RPMI1640 medium containing 10% fetal bovine serum at 5% CO2 and 37°C; (2) human liver cancer HepG2 cells, purchased from Beijing Solebow Company, cultured in MEM medium containing 10% fetal bovine serum at 5% CO2 and 37°C; (3) human lung cancer A549 cells, purchased from Beijing Solebow Company, cultured in F12-K medium containing 10% fetal bovine serum at 5% CO2 and 37°C; (4) human gastric cancer HGC-27 cells, purchased from Beijing Solebow Company, cultured in RPMI1640 medium containing 10% fetal bovine serum at 5% CO2 and 37°C; (5) human cervical cancer Hela cells, purchased from Beijing Solebow Company, cultured in RPMI1640 medium containing 10% fetal bovine serum at 5% CO2, 37℃; (6) Human normal liver L-02 cells, purchased from Shanghai Huiying Company, cultured in RPMI1640 medium containing 10% fetal bovine serum at 5% CO2, 37℃; (7) Human umbilical vein HUVEC cells, purchased from Shanghai Huiying Company, cultured in F12 medium containing 10% fetal bovine serum at 5% CO2, 37℃. According to the actual growth status of the cells, the cells were routinely subcultured according to the cell instructions, and the cells were used in experiments after being cultured for more than three generations and the growth status was stable.

[0033] Tumor cell survival rate assay: 100 µL was seeded in 96-well plates at a density of 1×10 5 Cells were cultured at 5% CO2 and 37°C for 12 h. 100 µL of medium containing different drug concentrations was added to each well. 5-Fluorouracil and resveratrol were used as positive controls. Six concentrations of 5, 10, 20, 50, 100, and 200 µM were set, and each concentration was repeated in triplicate. After drug treatment, 20 µL of 5 mg / mL thiazolyl blue (MTT) was added to each well for 4 h, and the supernatant was aspirated. 150 µL of DMSO was added and shaken for 5 min. The absorbance was detected at 570 nm on a microplate reader to calculate the cell survival rate (IC 50 ).

[0034] IC 50 The value represents the concentration of compound required to achieve 50% inhibition rate on tumor cells. The test results are shown in Table 1. Cell microscopy imaging after the action of different types and concentrations of drugs is shown in Table 1. Fig.11 shown.

[0035]

[0036] The results show that compounds I-1 to I-3 all have tumor cell killing effects, and have no killing effects on normal cells such as human liver cells; among them, compound I-1 has obvious killing activity on melanoma cells and lung cancer cells, compound I-2 has obvious killing activity on lung cancer cells and gastric cancer cells, and compound I-3 has obvious killing activity on melanoma cells, lung cancer cells, gastric cancer cells and cervical cancer cells. Although compound I-4 also shows killing activity on specific tumor cells, it also shows a strong killing effect on normal human liver cells, and has strong toxic side effects.

[0037] Unless otherwise specified, the scientific and technological terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. The present invention may also be implemented using any methods and materials similar to or equivalent to those described herein, and specific implementation methods and preferred methods and materials are described herein without limiting the present invention in any way.

Claims

1. An organic selenide, characterized in that Having the structure shown in I-1, I-2, I-3 or I-4: 。 2. The method for preparing the organic selenide according to claim 1, characterized in that: The following synthetic route was used to synthesize it: （1） ； （2） ； Among them, R Y represent , , or .

3. The preparation method according to claim 2, characterized in that: The step (1) comprises: taking selenium powder, mixing it with distilled water under nitrogen protection, slowly adding sodium borohydride aqueous solution while stirring, and after the system is clarified, adjusting the pH value of 2-chloroacetic acid to 9-10 with saturated sodium carbonate solution, adding it to the above reaction solution, and stirring at room temperature; collecting the reaction solution, adjusting the pH value to 2-3, extracting with ethyl acetate, collecting the organic phase, removing water, concentrating, and recrystallizing to obtain 2,2'-selenodiacetic acid.

4. The preparation method according to claim 2 or 3, characterized in that: The step (2) comprises: taking dichloromethane, adding N,N'-diisopropylethylamine, 1-hydroxy-7-azobenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 2,2'-selenodiacetic acid and raw material R Y -NH2, stir the reaction at room temperature for 10-15 h, then add water and stir the reaction for another 1-2 h. The resulting reaction solution was separated with a separatory funnel, the aqueous phase was collected, and dichloromethane was added for extraction. The organic phase was collected, dehydrated, and concentrated to obtain a crude product.

5. Use of the compound according to claim 1 in the preparation of drugs for preventing and / or treating cancer.

6. The use according to claim 5, characterized in that: The drug inhibits the proliferation of cancer cells, promotes apoptosis of cancer cells and / or inhibits the synthesis of cancer cell DNA.

7. The use according to claim 5 or 6, characterized in that: The compound has the structure shown in I-1, and the cancer is melanoma or lung cancer.

8. The use according to claim 5 or 6, characterized in that: The compound has the structure shown in I-2, and the cancer is lung cancer or gastric cancer.

9. The use according to claim 5 or 6, characterized in that: The compound has the structure shown in I-3, and the cancer is melanoma, lung cancer, gastric cancer or cervical cancer.

10. The use according to claim 5 or 6, characterized in that: The compound has the structure shown in I-4, and the cancer is melanoma, gastric cancer or cervical cancer.

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