Rhein-nitrogen mustard compound and application thereof in preparation of anti-cancer drugs

Abstract

The invention discloses a rhein-nitrogen mustard compound which is analyzed by X-ray single crystal diffraction, infrared spectrum, high-resolution mass spectrum and nuclear magnetic resonance spectrum structure and is a compound shown as a structural formula (I). Accordingly, the inventor also establishes a corresponding preparation method, namely the rhein is used as a main raw material and is subjected to esterification, acylation and hydrolysis reactions according to a reaction formula. In-vitro anticancer activity research shows that the rhein-nitrogen mustard compound has certain inhibitory activity and in-vitro anticancer activity on human lung cancer cells, human liver cancer cells, human breast cancer cells and human oral cancer cells, and has potential application value in research and development of anticancer drugs for human lung cancer, human liver cancer, human breast cancer and human oral cancer. In conclusion, the rhein-nitrogen mustard compound is good in anti-cancer activity, low in cost and simple in preparation method, and a new way is provided for developing new anti-cancer drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of rhein derivatives, and in particular relates to a rhein-nitrogen mustard compound and application thereof in the preparation of anticancer drugs. Background Art

[0002] Rhein is a bioactive ingredient extracted from traditional Chinese medicines such as rhubarb, Polygonum cuspidatum, and Guangxi regional specialty medicine Polygonum multiflorum. It is a natural anthraquinone compound and has the same parent nucleus structure as a variety of marketed anthraquinone anti-tumor drugs, such as mitoxantrone, daunorubicin, and epirubicin. Domestic and foreign studies have reported that rhein exhibits certain anti-tumor activity in both in vivo and in vitro experiments, and is therefore considered to be a potential anticancer drug and lead compound. Since the discovery of the anticancer activity of rhein, researchers have made a large number of structural modifications and designed and synthesized a large number of new rhein derivatives with different biological activities. Studies have shown that these rhein derivatives can achieve anti-tumor purposes through apoptosis, cycle arrest, para-apoptosis, autophagy, and radiosensitization. Therefore, the design and synthesis of new rhein derivatives with excellent anti-tumor activity has become a research direction of interest.

[0003] Nitrogen mustard anticancer drugs are an important branch of chemotherapy drugs and are widely used in the treatment of various malignant tumors in clinical practice. Since the use of nitrogen mustard as an alkylating agent in cancer chemotherapy in 1942, its various derivatives have been developed as drugs for the treatment of clinical cancer. For example, chlorambucil, cyclophosphamide and estramustine. Chlorambucil is mainly used to treat various malignant tumors such as chronic lymphocytic leukemia, ovarian cancer, and breast cancer. Cyclophosphamide is one of the most commonly used cytotoxic anticancer drugs, which works by producing active intermediates through oxidation in the body. In recent years, researchers have been working hard to develop new nitrogen mustard anticancer drugs to improve efficacy and reduce toxic side effects. Therefore, nitrogen mustard anticancer drugs have attracted much attention because of their ability to effectively inhibit the proliferation of cancer cells.

[0004] After investigation, the reports involving the application of rhein derivatives or nitrogen mustard derivatives as anticancer drugs are as follows:

[0005] Chinese patent "Preparation of rhein anilinoquinazoline derivatives and their application in anti-tumor drugs" (patent number CN201910261196.2, publication date 20190628).

[0006] Chinese patent application "Rhein-piperazine-sulfonamide hybrids and their preparation methods and anticancer drug applications" (patent application number CN2024102818009, publication date 20240614).

[0007] Chinese patent application "Rhein derivatives, preparation methods and applications in anti-cancer drugs" (patent application number CN202410235997.2, publication date 20240607).

[0008] Chinese patent application "Novel nitrogen mustard derivatives and preparation methods thereof" (patent application number CN202410394274.7, publication date 20240705).

[0009] Chinese patent "3-nitrogen mustard-2-sulfonyl acrylonitrile derivatives with anti-tumor activity and their preparation methods and applications" (patent number CN202311170634.7, publication date 20231212).

[0010] Literature (European journal of medicinal chemistry, 2019, 165, 31-45) reported that various rhein derivatives have inhibitory effects on human cervical cancer cells (HeLa), human colorectal cancer cells (HCT-116), human breast cancer cells (MCF-7), human promyelocytic leukemia cells (HL-60), human chronic myeloid leukemia cells (K562) and other cancer cells.

[0011] The literature (Bioorganic Chemistry.2023,135,106510) reports that a class of nitrogen mustard-quinazoline compounds exhibit excellent anti-cancer activity both in vivo and in vitro.

[0012] Despite this, there are no reports on rhein-nitrogen mustard compounds. Summary of the invention

[0013] The technical problem to be solved by the present invention is to provide a rhein-nitrogen mustard compound and its application in the preparation of anticancer drugs.

[0014] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0015] Rhein-nitrogen mustard compound, a compound of the following structural formula (I):

[0016]

[0017] The above-mentioned rhein-nitrogen mustard compound has a monoclinic crystal system and a space group of P2. 1 / n, α=90°, β=93.423(3)°, γ=90°, Z=4, Dc=1.570Mg·m -3 , F(000)=840.0.

[0018] The above-mentioned rhein-nitrogen mustard compound is a yellow needle-shaped crystal with a melting point of 183-184°C.

[0019] Application of the above rhein-nitrogen mustard compound in the preparation of anticancer drugs.

[0020] The cancer is lung cancer, liver cancer, breast cancer or oral cancer.

[0021] Lung cancer, liver cancer, breast cancer, and oral cancer are derived from human lung cancer cells A549, human liver cancer cells HepG2, human breast cancer cells MDA-MB-231, and human oral cancer cells CAL-27, respectively.

[0022] The preparation method of the above-mentioned rhein-nitrogen mustard compound is carried out using rhein as the main raw material according to the following reaction formula:

[0023] Wherein, R is selected from ethyl, propyl or butyl; when R is ethyl, the intermediate is A1, B1, C1; when R is propyl, the intermediate is A2, B2, C2; when R is butyl, the intermediate is A3, B3, C3.

[0024] The above preparation method comprises the following steps:

[0025] (a) Dissolve 1 mmol of rhein in 30 ml of anhydride at room temperature, slowly add 0.5 ml of concentrated sulfuric acid dropwise under stirring at 0°C, then heat to 30°C and stir for 4 hours to obtain intermediate A;

[0026] (b) Under nitrogen protection, 10 ml of anhydrous dichloromethane, 0.2 ml of oxalyl chloride, and 1 drop of N,N-dimethylformamide were added to a flask, and the mixture was stirred at 30°C for 15 min. Then, 1 mmol of intermediate A was added, and the mixture was stirred at 30°C for 60 min to obtain intermediate B;

[0027] (c) 1 mmol of bis(2-chloroethyl)amine hydrochloride and 2 mmol of anhydrous potassium carbonate were suspended in 30 ml of dichloromethane at room temperature, stirred for 10 min, and then 1 mmol of intermediate B was slowly added dropwise. The mixture was stirred for 2 h at 30° C. to obtain intermediate C;

[0028] (d) 1 mmol of intermediate C was suspended in ethanol:water (V / V=3:1) at room temperature, an inorganic or organic base was added, the mixture was reacted at room temperature for 10 h, and the pH was adjusted to 2-7 with dilute hydrochloric acid.

[0029] In step (a), the acid anhydride is acetic anhydride, propionic anhydride or butyric anhydride; in the case of acetic anhydride, the corresponding intermediates A, B and C are A1, B1 and C1 respectively; in the case of propionic anhydride, the corresponding intermediates A, B and C are A2, B2 and C2 respectively; in the case of butyric anhydride, the corresponding intermediates A, B and C are A3, B3 and C3 respectively.

[0030] In step (d), the inorganic base is potassium hydroxide, sodium hydroxide, potassium carbonate or sodium carbonate, and the organic base is triethylamine.

[0031] Based on the previous research on rhein derivatives, the inventors designed and prepared a rhein-nitrogen mustard compound, which was analyzed by X-ray single crystal diffraction, infrared spectroscopy, high-resolution mass spectrometry, and nuclear magnetic resonance spectroscopy to be a compound of the following structural formula (I):

[0032]

[0033] Based on this, the inventors have also established a corresponding preparation method, that is, using rhein as the main raw material and performing esterification, acylation, and hydrolysis according to the reaction formula. In vitro anticancer activity studies have shown that the rhein-nitrogen mustard compound of the present invention has certain inhibitory activity and in vitro anticancer activity against human lung cancer cells, human liver cancer cells, human breast cancer cells, and human oral cancer, and has potential application value in the research and development of anticancer drugs for human lung cancer, human liver cancer, human breast cancer, and human oral cancer. In summary, the rhein-nitrogen mustard compound of the present invention has good anticancer activity, low cost, and a simple preparation method, providing a new approach for the development of new anticancer drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The figure is a crystal structure diagram of the rhein-nitrogen mustard compound of the present invention.

[0035] Figure 2 The rhein-nitrogen mustard compound of the present invention 1 H NMR spectrum.

[0036] Figure 3 The rhein-nitrogen mustard compound of the present invention 13 C NMR spectrum.

[0037] Figure 4 It is the HRMS spectrum of the rhein-nitrogen mustard compound of the present invention. DETAILED DESCRIPTION

[0038] Example 1 Preparation of Intermediate A1

[0039] Dissolve 1 mmol of rhein in 30 ml of acetic anhydride at room temperature, slowly add 0.5 ml of concentrated sulfuric acid under stirring at 0°C, then heat to 30°C and stir for 4 hours. Stop the reaction, pour the reaction solution into 200 ml of ice water, stir rapidly, precipitate a yellow solid, filter, wash the filter cake with ice water, and vacuum dry to obtain a yellow solid with a yield of 91%.

[0040] Example 2 Preparation of Intermediate A2

[0041] Dissolve 1 mmol of rhein in 30 ml of propionic anhydride at room temperature, slowly add 0.5 ml of concentrated sulfuric acid under stirring at 0°C, then heat to 30°C and stir for 4 hours. Stop the reaction, pour the reaction solution into 200 ml of ice water, stir rapidly, precipitate a yellow solid, filter, wash the filter cake with ice water, and vacuum dry to obtain a yellow solid with a yield of 86%.

[0042] Example 3 Preparation of Intermediate A3

[0043] Dissolve 1 mmol of rhein in 30 ml of butyric anhydride at room temperature, slowly add 0.5 ml of concentrated sulfuric acid under stirring at 0°C, then heat to 30°C and stir for 4 hours. Stop the reaction, pour the reaction solution into 200 ml of ice water, stir rapidly, precipitate a yellow solid, filter, wash the filter cake with ice water, and vacuum dry to obtain a yellow solid with a yield of 85%.

[0044] Example 4 Preparation of Intermediate B1

[0045] Under nitrogen protection, add 10 ml of anhydrous dichloromethane, 0.2 ml of oxalyl chloride, and 1 drop of N,N-dimethylformamide to the flask, stir and react at 30°C for 15 minutes, then add 1 mmol of intermediate A1, stir and react at 30°C for 60 minutes. Remove all solvents under reduced pressure to obtain intermediate B1 as a yellow solid with a yield of 100%. Dissolve intermediate B1 in 5 ml of anhydrous dichloromethane for later use.

[0046] Example 5 Preparation of Intermediate B2

[0047] Under nitrogen protection, add 10 ml of anhydrous dichloromethane, 0.2 ml of oxalyl chloride, and 1 drop of N,N-dimethylformamide to the flask, stir and react at 30°C for 15 min, then add 1 mmol of intermediate A2, stir and react at 30°C for 60 min. Remove all solvents under reduced pressure to obtain intermediate B1 as a yellow solid with a yield of 100%. Dissolve intermediate B2 in 5 ml of anhydrous dichloromethane for later use.

[0048] Example 6 Preparation of Intermediate B3

[0049] Under nitrogen protection, add 10 ml of anhydrous dichloromethane, 0.2 ml of oxalyl chloride, and 1 drop of N,N-dimethylformamide to the flask, stir and react at 30°C for 15 minutes, then add 1 mmol of intermediate A3, stir and react at 30°C for 60 minutes. Remove all solvents under reduced pressure to obtain intermediate B1 as a yellow solid with a yield of 100%. Dissolve intermediate B3 in 5 ml of anhydrous dichloromethane for later use.

[0050] Example 7 Preparation of Intermediate C1

[0051] At room temperature, 1 mmol of bis(2-chloroethyl)amine hydrochloride and 2 mmol of anhydrous potassium carbonate were suspended in 30 ml of dichloromethane, stirred for 10 min, and then 1 mmol of intermediate B1 was slowly added dropwise, and stirred for 2 h at 30°C. The mixture was filtered, and the filter cake was washed with dichloromethane. All solvents were removed under reduced pressure to obtain intermediate C1 as a yellow solid with a yield of 95%.

[0052] Example 8 Preparation of Intermediate C2

[0053] At room temperature, 1 mmol of bis(2-chloroethyl)amine hydrochloride and 2 mmol of anhydrous potassium carbonate were suspended in 30 ml of dichloromethane, stirred for 10 min, and then 1 mmol of intermediate B2 was slowly added dropwise, and stirred for 2 h at 30°C. The mixture was filtered, and the filter cake was washed with dichloromethane. All solvents were removed under reduced pressure to obtain intermediate C2 as a yellow solid with a yield of 96%.

[0054] Example 9 Preparation of Intermediate C3

[0055] At room temperature, 1 mmol of bis(2-chloroethyl)amine hydrochloride and 2 mmol of anhydrous potassium carbonate were suspended in 30 ml of dichloromethane, stirred for 10 min, and then 1 mmol of intermediate B3 was slowly added dropwise, and stirred for 2 h at 30°C. The mixture was filtered, and the filter cake was washed with dichloromethane. All solvents were removed under reduced pressure to obtain intermediate C3 as a yellow solid with a yield of 92%.

[0056] Example 10 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0057] At room temperature, 1 mmol of intermediate C1 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of potassium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 2 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 83%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0058] The results of X-ray single crystal diffraction, infrared spectroscopy, high-resolution mass spectrometry, and nuclear magnetic resonance spectroscopy structural analysis are as follows:

[0059] Crystallographic data: monoclinic, space group P2 1 / n, α=90°, β=93.423(3)°, γ=90°, Z=4, Dc=1.570Mg·m -3 ,μ(Cu F(000)=840.0.

[0060] Infrared spectrum data: FT-IR (KBr, ν / cm -1 ):3434,3120,2359,2341,1680,1630,1570,1553,1468,1452,1401,1375,1354,1269,1246,1194,1163,1149,1083,1056,754,668.

[0061] H NMR spectrum data: 1 H NMR (500 MHz, CDCl 3 )δ12.11(s,1H),11.97(s,1H),7.95–7.81(m,2H),7.72(t,J=8.0Hz,1H),7.34(dd,J=4.4,3.0Hz,2H),3.93–3.74(m,6H),3.57(s,2H).

[0062] NMR carbon spectrum data: 13 C NMR (126 MHz, CDCl 3 )δ192.65,180.96,169.77,162.81,162.66,144.32,137.67,134.27,133.43, 125.04,122.58,120.42,117.88,116.45,115.76,51.78,47.70,41.35,41.21.

[0063] High resolution mass spectrometry data: HRMS (ESI) m / z [M+Na] + Calculate for C 19 H 15 NO 5 Cl 2 Na + :430.0222,found:430.0225.

[0064] The structural analysis results of Examples 11 to 22 are the same as those of Example 10 and will not be repeated here.

[0065] Example 11 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0066] At room temperature, 1 mmol of intermediate C1 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of sodium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 7 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 81%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0067] Example 12 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0068] At room temperature, 1 mmol of intermediate C1 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of potassium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 5 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 85%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0069] Example 13 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0070] At room temperature, 1 mmol of intermediate C1 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of sodium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 6 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 83%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0071] Example 14 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0072] At room temperature, 1 mmol of intermediate C2 was suspended in ethanol: water (V / V = 3: 1), and 2.2 mmol of potassium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 3 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 79%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184 ° C.

[0073] Example 15 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0074] At room temperature, 1 mmol of intermediate C2 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of sodium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 4 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 81%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0075] Example 16 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0076] At room temperature, 1 mmol of intermediate C2 was suspended in ethanol: water (V / V = 3: 1), and 2.2 mmol of potassium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 6 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 84%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0077] Example 17 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0078] At room temperature, 1 mmol of intermediate C2 was suspended in ethanol: water (V / V = 3: 1), and 2.2 mmol of sodium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 2 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 86%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184 ℃.

[0079] Example 18 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0080] At room temperature, 1 mmol of intermediate C3 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of potassium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 7 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 78%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0081] Example 19 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0082] At room temperature, 1 mmol of intermediate C3 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of sodium hydroxide was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 5 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 76%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0083] Example 20 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0084] At room temperature, 1 mmol of intermediate C3 was suspended in ethanol: water (V / V = 3: 1), and 2.2 mmol of potassium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 4 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 81%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0085] Example 21 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0086] At room temperature, 1 mmol of intermediate C3 was suspended in ethanol: water (V / V = 3:1), and 2.2 mmol of sodium carbonate was added. The reaction was carried out at room temperature for 10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 2 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 83%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0087] Example 22 Preparation of Rhein-Nitrogen Mustard Compound (I)

[0088] At room temperature, 1 mmol of intermediate C3 was suspended in ethanol: water (V / V = 3: 1), and 2.2 mmol of triethylamine was added. The reaction was carried out at room temperature for 8-10 hours. The solution turned purple-red, and the reaction was stopped. The pH was adjusted to 3 with dilute hydrochloric acid, and a large amount of yellow solid precipitated. Filter, wash the filter cake with ice water, and vacuum dry to obtain a crude product. Purify the product by silica gel column chromatography to obtain a pure product, a yellow solid, with a yield of 87%. Use methanol + dichloromethane to slowly evaporate the solvent at room temperature to obtain yellow needle-shaped crystals. Mp: 183-184℃.

[0089] Application example: MTT method to detect the effect of compounds on tumor cell proliferation

[0090] Lung cancer (A549), liver cancer (HepG2), breast cancer (MDA-MB-231) and oral cancer (CAL-27) cells in the logarithmic growth phase were cultured at 3×10 3 The cells were inoculated at a density of 100 μL / well in a 96-well culture plate. After the cells adhered to the wall for 24 h, the original culture medium was carefully discarded. 100 μL of culture medium was added to the control group, and 100 μL of different concentrations of the compound obtained in Example 10 (abbreviated as RNC) were added to the experimental groups. Three parallel wells were set for each concentration. The plates were placed at 37°C and 5% CO. 2 The culture was continued in the incubator for 48 h, and 5 mg·L -1 Add 20 μL of MTT solution, continue incubation for 4 hours, carefully discard the supernatant, add 100 μL DMSO to each well to dissolve, shake at low speed for 10 minutes, measure the absorbance value (OD value) of each well at a wavelength of 490 nm on an automatic microplate reader, and calculate the cell growth inhibition rate according to the following formula: Cell growth inhibition rate % = [1-(OD value of drug-added group / OD value of blank group)] × 100%.

[0091] Table 1 Effects of compounds on proliferation of different cancer cells

[0092]

[0093] The rhein-nitrogen mustard compounds prepared in other Examples 11 to 22 were tested for anticancer activity in the same manner as in the application example, and the test results were similar to those in Table 1 and are not described in detail.

Claims

1. A rhein-nitrogen mustard compound, characterized in that It is a compound of the following structural formula (I):

2. The rhein-nitrogen mustard compound according to claim 1, characterized in that Its crystal is monoclinic, space group P21 / n, α=90°,β=93.423(3)°, γ=90°,Z=4, Dc=1.570Mg·m -3 , F(000)=840.0。 3. The rhein-nitrogen mustard compound according to claim 2, characterized in that It is yellow needle-shaped crystals with a melting point of 183-184°C.

4. Use of the rhein-nitrogen mustard compound according to claim 1 in the preparation of anticancer drugs.

5. The use according to claim 4, characterized in that: The cancer is lung cancer, liver cancer, breast cancer or oral cancer.

6. The use according to claim 5, characterized in that: The lung cancer, liver cancer, breast cancer and oral cancer are derived from human lung cancer cells A549, human liver cancer cells HepG2, human breast cancer cells MDA-MB-231 and human oral cancer cells CAL-27, respectively.

7. The method for preparing the rhein-nitrogen mustard compound according to claim 1, characterized in that With rhein as the main raw material, the process is carried out through esterification, acylation and hydrolysis according to the following reaction formula: Wherein, R is selected from ethyl, propyl or butyl; when R is ethyl, the intermediate is A1, B1, C1; when R is propyl, the intermediate is A2, B2, C2; when R is butyl, the intermediate is A3, B3, C3.

8. The preparation method according to claim 7, characterized in that The following steps are involved: (a) Dissolve 1 mmol of rhein in 30 ml of anhydride at room temperature, slowly add 0.5 ml of concentrated sulfuric acid dropwise under stirring at 0°C, then heat to 30°C and stir for 4 hours to obtain intermediate A; (b) Under nitrogen protection, 10 ml of anhydrous dichloromethane, 0.2 ml of oxalyl chloride, and 1 drop of N,N-dimethylformamide were added to a flask, and the mixture was stirred at 30°C for 15 min. Then, 1 mmol of intermediate A was added, and the mixture was stirred at 30°C for 60 min to obtain intermediate B; (c) 1 mmol of bis(2-chloroethyl)amine hydrochloride and 2 mmol of anhydrous potassium carbonate were suspended in 30 ml of dichloromethane at room temperature, stirred for 10 min, and then 1 mmol of intermediate B was slowly added dropwise. The mixture was stirred for 2 h at 30° C. to obtain intermediate C; (d) 1 mmol of intermediate C was suspended in ethanol:water (V / V=3:1) at room temperature, an inorganic or organic base was added, the mixture was reacted at room temperature for 10 h, and the pH was adjusted to 2-7 with dilute hydrochloric acid.

9. The preparation method according to claim 7, characterized in that: In step (a), the acid anhydride is acetic anhydride, propionic anhydride or butyric anhydride; in the case of acetic anhydride, the corresponding intermediates A, B and C are A1, B1 and C1 respectively; in the case of propionic anhydride, the corresponding intermediates A, B and C are A2, B2 and C2 respectively; in the case of butyric anhydride, the corresponding intermediates A, B and C are A3, B3 and C3 respectively.

10. The preparation method according to claim 7, characterized in that: In step (d), the inorganic base is potassium hydroxide, sodium hydroxide, potassium carbonate or sodium carbonate, and the organic base is triethylamine.

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