Organic selenium compounds and their use in preparing anticancer drugs

Abstract

The present invention provides novel organoselenium compounds and methods for their synthesis, as well as their use in the preparation of drugs for the prevention and / or treatment of cancer. The compounds provided by the present invention combine naturally occurring active compounds with selenium, creating a synergistic anticancer effect and enhancing their safety and anticancer efficacy, showing great potential for the prevention and treatment of cancer.

Description

Technical Field

[0001] The present invention belongs to the field of drug synthesis, and in particular 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, with the number of cancer patients continuing to rise annually. While significant progress has been made in cancer drug treatments, including chemotherapy and targeted therapies, many challenges and pain points remain. Chemotherapeutic drugs exhibit nonspecific toxicity, have relatively limited clinical targets, and targeted therapies have relatively narrow indications. Similar to chemotherapy, targeted therapies are prone to developing drug resistance.

[0003] Selenium is an essential trace element in living organisms, possessing numerous unique functions for human health. It is known as the "spark of life" and the "king of cancer prevention." Selenium is the active center of glutathione peroxidase (GSH-Px) and possesses potent antioxidant properties. Selenium deficiency in the human body can lead to the development of dozens of diseases. my country has made significant progress in preventing Keshan disease, Kashin-Beck disease, and liver cancer through selenium supplementation. Selenium's anti-cancer effects are primarily achieved through various mechanisms, including antioxidant activity, immunomodulatory effects, inhibition of cancer cell proliferation and angiogenesis, and promotion of DNA repair. However, most of the selenium used in clinical practice is inorganic, such as sodium selenite and selenium disulfide. These inorganic selenium products suffer from significant toxic side effects and low bioavailability. Currently, a few organic selenium drugs, such as ebselen, are in clinical trials, leaving few promising candidates for clinical anti-cancer use. Summary of the Invention

[0004] To overcome the shortcomings in the existing field, the present invention synthesizes a series of organic selenium compounds with cancer cell killing properties by combining selenium with o-methoxybenzylamine through reduction, etherification, amidation and other reactions. 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] .

[0007] The compound provided by the present invention combines selenium with aniline or benzylamine traditional Chinese medicine monomers through chemical bonds to form a new selenium-containing active molecule, so that the two can exert a synergistic effect.

[0008] In a second aspect, the present invention provides a method for preparing the compound described in the first aspect.

[0009] The organic selenium compound of the present invention is prepared by the following synthetic route:

[0010] (1) ;

[0011] (2) ;

[0012] Among them, R Y represent 、 、 or .

[0013] 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 2-chloroacetic acid to a pH value of 9-10 with a saturated sodium carbonate solution, adding the solution 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.

[0014] 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 at room temperature for 10-15 h, then add water and stir for another 1-2 h. The resulting reaction solution is separated with a separatory funnel, the aqueous phase is collected, and dichloromethane is added for extraction. The organic phase is collected, dehydrated, and concentrated to obtain a crude product.

[0015] 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.

[0016] 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.

[0017] 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.

[0018] 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.

[0019] 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.

[0020] 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.

[0021] As a preferred embodiment of the present invention, the drug exerts its anti-cancer effect by inhibiting the proliferation of cancer cells, promoting the apoptosis of cancer cells, and inhibiting the synthesis of DNA through one or more mechanisms.

[0022] Compared with the existing technology, 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, so that the anti-cancer activities of the two produce a synergistic effect, and increase its safety and anti-cancer effect, which has great potential in the prevention and treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 、Compound 1 1 H NMR spectrum (600 MHz, D O);

[0024] Figure 2 , ESI-MS spectrum of compound 1;

[0025] Figure 3 、Compound I-1 1 H NMR spectrum (600 MHz, DMSO- d 6);

[0026] Figure 4 , ESI-MS spectrum of compound I-1;

[0027] Figure 5 , Compound I-2 1 H NMR spectrum (600 MHz, DMSO- d 6);

[0028] Figure 6 , ESI-MS spectrum of compound I-2;

[0029] Figure 7 , Compound I-3 1 H NMR spectrum (600 MHz, DMSO- d 6);

[0030] Figure 8 , ESI-MS spectrum of compound I-3;

[0031] Figure 9 , Compound I-4 1 H NMR spectrum (600 MHz, DMSO- d 6);

[0032] Figure 10 , ESI-MS spectrum of compound I-4;

[0033] Figure 11 , B16 cells, Hela cells, HGC-27 cells and L-O2 cells under different culture conditions. DETAILED DESCRIPTION

[0034] 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 the present invention is limited solely by the claims. Those skilled in the art may make various modifications and improvements to various aspects of the present invention without departing from the scope of the claims, and such modifications and improvements are also within the scope of the present invention.

[0035] 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 well known to those skilled in the art.

[0036] Example 1: Synthesis of Compound I-1

[0037] Compound I-1 is 2,2'-selenobis(N-benzylacetamide) with the following structure:

[0038]

[0039] Compound I-1 was synthesized by the following method:

[0040] (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 add 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 and adjust the pH 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 complete, filter the reaction solution, adjust the pH of the filtrate to 2-3 with dilute hydrochloric acid, extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate and crystallize, and then recrystallize from 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 shown).

[0041] (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. Stir at room temperature for 12 h. Add 20 ml of water and stir for 1 h. Remove the stirrer, separate the mixture in a separatory funnel, extract with DCM, wash with water, dry, and rotary evaporate the DCM to obtain a crude product. The crude product was eluted with a semi-preparative HPLC gradient of acetonitrile / water (volume ratio 20:70 to 100:0) to obtain 0.289 g of the purified product I-1 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 shown).

[0042] Example 2: Synthesis of Compound I-2

[0043] Compound I-2 is 2,2'-selenobis(N-(2-methoxyphenyl)acetamide) with the following structure:

[0044]

[0045] Using the method provided in Example 1, except that 0.214 g (0.002 mol) of benzylamine was replaced with 0.246 g (0.002 mol) of o-anisidine in step (2), 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 shown).

[0046] Example 3: Synthesis of Compound I-3

[0047] Compound I-3 is 2,2'-selenobis(N-(2-methoxybenzyl)acetamide) with the following structure:

[0048]

[0049] Using the method provided in Example 1, except that 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 shown).

[0050] Example 4: Synthesis of Compound I-4

[0051] Compound I-4 is 2,2'-selenobis(N-(3-methoxybenzyl)acetamide) with the following structure:

[0052]

[0053] Using the method provided in Example 1, except that 0.214 g (0.002 mol) of benzylamine was replaced with 0.274 g (0.002 mol) of m-methoxybenzylamine in step (2), 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. Figure 9 ESI-MS( m / z ): 435.0891[MH] - (like Figure 10 shown).

[0054] Experimental example

[0055] This experimental example verifies the anti-tumor activity of compounds I-1 to I-4, using resveratrol and 5-fluorouracil as controls.

[0056] 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% (6) Human normal liver L-02 cells were purchased from Shanghai Huiying Company and cultured in RPMI1640 medium containing 10% fetal bovine serum at 5% CO2 and 37°C. (7) Human umbilical vein HUVEC cells were purchased from Shanghai Huiying Company and cultured in F12 medium containing 10% fetal bovine serum at 5% CO2 and 37°C. Cells were routinely passaged according to the instructions of the cell manufacturer based on their actual growth status. Cells were used in experiments after they had been cultured for more than three generations and their growth status was stable.

[0057] Tumor cell survival rate assay: 100 μL of cells were seeded in 96-well plates at a density of 1×10 5 Cells were cultured at 5% CO2 and 37°C for 12 hours. 100 µL of culture 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, with three replicates for each concentration. After drug treatment, 20 µL of 5 mg / mL MTT was added to each well for 4 hours. The supernatant was aspirated, 150 µL of DMSO was added, and the cells were shaken for 5 minutes. The absorbance was measured at 570 nm on a microplate reader to calculate the cell survival rate (IC). 50 ).

[0058] IC 50 The values ​​represent 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. Figure 11 shown.

[0059]

[0060] 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 hepatocytes. Among them, compound I-1 has significant killing activity against melanoma cells and lung cancer cells, compound I-2 has significant killing activity against lung cancer cells and gastric cancer cells, and compound I-3 has significant killing activity against melanoma cells, lung cancer cells, gastric cancer cells, and cervical cancer cells. Although compound I-4 also exhibits killing activity against specific tumor cells, it also shows a strong killing effect on normal human hepatocytes and has strong toxic side effects.

[0061] Unless otherwise indicated, the scientific and technical terms used herein have the same meanings as those commonly 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 or equivalent to those described herein. Specific embodiments and preferred methods and materials are described herein and are not intended to limit the present invention in any way.

Claims

1. Use of an organic selenium compound in the preparation of a drug for preventing and / or treating melanoma or cervical cancer, wherein the organic selenium compound has the structure shown below:

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