Rhein-piperazine-furanone hybrid and its preparation method and application

Abstract

The present invention provides a rhein-piperazine-furanone hybrid and a preparation method and application thereof, and relates to the technical field of rhein pharmaceutical chemistry. The present invention first prepares compound 4, and then reacts with an acyl chloride intermediate 3 of rhein 1 or diacerein 2. N The rhein-piperazine-furanone hybrid 5 is obtained by the acylation reaction, the synthesis route is simple, the by-products are few, the product separation and purification operation is simple, and the reaction yield is above 48%. The rhein-piperazine-furanone hybrid 5 prepared by the present invention has strong toxicity to various cancer cells and low toxicity to normal cells, and has excellent anticancer activity and selectivity.

Description

Technical Field

[0001] The invention relates to the technical field of rhein pharmaceutical chemistry, and in particular to a rhein-piperazine-furanone hybrid and a preparation method and application thereof. Background Art

[0002] Cancer is increasingly becoming a leading cause of death in countries around the world. It is estimated that 19.3 million new cancer cases occurred in 2020, resulting in 10 million deaths from cancer worldwide. Chemotherapy with drugs is an important means of treating cancer, and more than 60% of anticancer agents are natural product derivatives. Natural products are an important source of clinical drugs. Among the 371 drugs in the ninth edition of the International Pharmacopoeia, more than 80 are natural products and their derivatives.

[0003] Rhein is an anthraquinone natural product extracted from medicinal plants such as rhubarb, cassia seed, Polygonum multiflorum and aloe vera. Plants containing rhein are used to prevent and treat different diseases in different traditional medical systems. As early as more than 1,000 years ago, people began to use rhubarb or rhubarb components as laxatives. However, the poor bioavailability, low water solubility and easy gastrointestinal diseases of rhein have hindered its clinical application. In the prior art, the Chinese invention patent with application number: 202210453596.5 discloses a rhein-piperazine-dithiocarbamate hybrid and its preparation method and application, and specifically discloses a compound as shown in formula (I) or its pharmaceutically acceptable salt, isotope label, solvate, polymorph or prodrug. The invention obtains a series of rhein-piperazine-dithiocarbamate hybrids by hybridizing rhein derivatives with dithiocarbamate groups. Experiments have shown that the hybrids inhibit the growth of various cancer cells and have low toxicity to normal cells. However, the invention has limited improvement in the biological activity and selectivity of rhein, resulting in low efficacy.

[0004] Summary of the invention

[0005] In view of this, the present invention provides a rhein-piperazine-furanone hybrid which improves the biological activity and selectivity of rhein.

[0006] It is also necessary to provide a method for preparing rhein-piperazine-furanone hybrids.

[0007] It is also necessary to provide an application of rhein-piperazine-furanone hybrids.

[0008] The technical solution adopted by the present invention to solve its technical problem is:

[0009] A rhein-piperazine-furanone hybrid, wherein the general structural formula of the rhein-piperazine-furanone hybrid is as follows:

[0010]

[0011] in,

[0012] X is a halogen atom,

[0013] R 1 is selected from hydrogen or acyl,

[0014] R 2 Selected from hydrocarbon groups or hydrocarbon groups containing heteroatoms.

[0015] Preferably, X is a chlorine atom or a bromine atom, and R 1 is selected from hydrogen or acetyl, said R 2 Selected from methyl, menthyl or bornyl.

[0016] A method for preparing a rhein-piperazine-furanone hybrid, comprising the following steps:

[0017] S1: Mix mucochloric acid or mucobromic acid with R 2 OH undergoes etherification reaction in catalyst A and aprotic solvent to obtain intermediate 4-1, and intermediate 4-1 undergoes nucleophilic substitution reaction with piperazine in acid-binding agent B and polar aprotic solvent to obtain compound 4;

[0018] S2: Rhein (1) or diacerein (2) is subjected to chlorination reaction with oxalyl chloride in a catalyst C and a polar aprotic solvent to obtain an acyl chloride intermediate 3; intermediate 3 is subjected to N-acylation reaction with compound 4 in an acid-binding agent B and a polar aprotic solvent to obtain a rhein-piperazine-furanone hybrid 5;

[0019] The synthetic route is as follows:

[0020]

[0021] Preferably, the R 2 OH is methanol or menthol or borneol;

[0022] The catalyst A is concentrated sulfuric acid;

[0023] The acid binding agent B is triethylamine;

[0024] The catalyst C is N,N-dimethylformamide;

[0025] The aprotic solvent is 1,2-dichloroethane or cyclohexane;

[0026] The polar aprotic solvent is dichloromethane.

[0027] A pharmaceutically acceptable salt of a rhein-piperazine-furanone hybrid as described above.

[0028] A pharmaceutical composition of the rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof as described above.

[0029] The pharmaceutical preparation of the rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof as described above further comprises at least one pharmaceutically acceptable excipient or carrier.

[0030] The pharmaceutical preparation of the above-mentioned rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof is a tablet, a capsule, a granule or an injection.

[0031] Use of the above-mentioned rhein-piperazine-furanone hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation in the preparation of anticancer drugs.

[0032] Use of the above-mentioned rhein-piperazine-furanone hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation in the preparation of drugs against lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] 1. The present invention first prepares compound 4, and then carries out N-acylation reaction with an acyl chloride intermediate 3 of rhein 1 or diacerein 2 in an acid-binding agent B and a polar aprotic solvent to obtain a product rhein-piperazine-furanone hybrid 5. The synthetic route is simple, the by-products are few, the product separation and purification operation is simple, and the reaction yield is above 48%.

[0035] 2. The rhein-piperazine-furanone hybrid prepared by the present invention exhibits good cytotoxicity to various cancer cells and less toxicity to normal cells, and has excellent anticancer activity and selectivity. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 for the rhein-piperazine-furanone hybrid 5a 1 H NMR spectrum.

[0037] Figure 2 for the rhein-piperazine-furanone hybrid 5a 13 C NMR spectrum.

[0038] Figure 3 HRMS spectrum of rhein-piperazine-furanone hybrid 5a.

[0039] Figure 4 for the rhein-piperazine-furanone hybrid 5b 1 HNMR spectrum.

[0040] Figure 5 for the rhein-piperazine-furanone hybrid 5b 13 C NMR spectrum.

[0041] Figure 6 HRMS spectrum of rhein-piperazine-furanone hybrid 5b.

[0042] Figure 7 for the rhein-piperazine-furanone hybrid 5c 1 H NMR spectrum.

[0043] Figure 8 for the rhein-piperazine-furanone hybrid 5c 13 C NMR spectrum.

[0044] Fig. 9 HRMS spectrum of rhein-piperazine-furanone hybrid 5c.

[0045] Fig.10 for the rhein-piperazine-furanone hybrid 5d 1 HNMR spectrum.

[0046] Fig.11 for the rhein-piperazine-furanone hybrid 5d 13 C NMR spectrum.

[0047] Fig.12 This is the HRMS spectrum of rhein-piperazine-furanone hybrid 5d.

[0048] Fig.13 for the rhein-piperazine-furanone hybrid 5e 1 H NMR spectrum.

[0049] Fig.14 for the rhein-piperazine-furanone hybrid 5e 13 C NMR spectrum.

[0050] Fig.15 HRMS spectrum of rhein-piperazine-furanone hybrid 5e.

[0051] Fig.16 for the rhein-piperazine-furanone hybrid 5f 1 H NMR spectrum.

[0052] Fig.17 for the rhein-piperazine-furanone hybrid 5f 13C NMR spectrum.

[0053] Fig.18 HRMS spectrum of rhein-piperazine-furanone hybrid 5f.

[0054] Fig.19 5g of rhein-piperazine-furanone hybrid 1 HNMR spectrum.

[0055] Fig. 20 5g of rhein-piperazine-furanone hybrid 13 C NMR spectrum.

[0056] Fig.21 This is the HRMS spectrum of 5g of rhein-piperazine-furanone hybrid.

[0057] Fig. 22 Rhein-piperazine-furanone hybrid 5h 1 HNMR spectrum.

[0058] Fig.23 Rhein-piperazine-furanone hybrid 5h 13 C NMR spectrum.

[0059] Fig.24 This is the HRMS spectrum of rhein-piperazine-furanone hybrid 5h.

[0060] Fig.25 for the rhein-piperazine-furanone hybrid 5i 1 HNMR spectrum.

[0061] Fig.26 for the rhein-piperazine-furanone hybrid 5i 13 C NMR spectrum.

[0062] Fig. 27 HRMS spectrum of rhein-piperazine-furanone hybrid 5i.

[0063] Fig.28 for the rhein-piperazine-furanone hybrid 5j 1 HNMR spectrum.

[0064] Fig.29 for the rhein-piperazine-furanone hybrid 5j 13 C NMR spectrum.

[0065] Fig.30 HRMS spectrum of rhein-piperazine-furanone hybrid 5j.

[0066] Fig.31The graph shows the inhibitory effects of rhein-piperazine-furanone hybrids 5a-5j, Rhein and cytarabine (CAR) on A549 and WI-38 cell lines.

[0067] Fig.32 This is a diagram of H460 cell apoptosis induced by rhein-piperazine-furanone hybrid 5e; in the figure: (A) flow cytometric analysis of H460 cells; (B) total apoptosis rate of H460 cells. DETAILED DESCRIPTION

[0068] The technical solutions and technical effects of the embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings of the present invention.

[0069] A rhein-piperazine-furanone hybrid, wherein the general structural formula of the rhein-piperazine-furanone hybrid is as follows:

[0070]

[0071] X is a halogen atom,

[0072] R 1 is selected from hydrogen or acyl,

[0073] R 2 Selected from hydrocarbon groups or hydrocarbon groups containing heteroatoms.

[0074] Furthermore, X is a chlorine atom or a bromine atom, and R 1 is selected from hydrogen or acetyl, said R 2 Selected from methyl, menthyl or bornyl.

[0075] Compared with the prior art, the present invention has the following beneficial effects:

[0076] 1. The present invention first prepares compound 4, and then carries out N-acylation reaction with an acyl chloride intermediate 3 of rhein 1 or diacerein 2 in an acid-binding agent B and a polar aprotic solvent to obtain a product rhein-piperazine-furanone hybrid 5. The synthetic route is simple, the by-products are few, the product separation and purification operation is simple, and the reaction yield is above 48%.

[0077] 2. The rhein-piperazine-furanone hybrid prepared by the present invention exhibits good cytotoxicity to various cancer cells and less toxicity to normal cells, and has excellent anticancer activity and selectivity.

[0078] A method for preparing a rhein-piperazine-furanone hybrid, comprising the following steps:

[0079] S1: Mix mucochloric acid or mucobromic acid with R 2OH undergoes etherification reaction in catalyst A and aprotic solvent to obtain intermediate 4-1, and intermediate 4-1 undergoes nucleophilic substitution reaction with piperazine in acid-binding agent B and polar aprotic solvent to obtain compound 4;

[0080] S2: Rhein (1) or diacerein (2) is subjected to chlorination reaction with oxalyl chloride in a catalyst C and a polar aprotic solvent to obtain an acyl chloride intermediate 3; intermediate 3 is subjected to N-acylation reaction with compound 4 in an acid-binding agent B and a polar aprotic solvent to obtain a rhein-piperazine-furanone hybrid 5;

[0081] Specifically, the diacerein can be purchased or prepared by the following method: at 0°C, add concentrated sulfuric acid (0.88 mL, 11 mmol) slowly (about 1 h to complete the dripping) to a mixture of rhein 1 (1.42 g, 5 mmol) and acetic anhydride (1.53 g, 15 mmol), heat to 35°C and stir for 3 h. The above reaction solution is slowly dripped into 0°C water (100 mL), and the temperature in the bottle is controlled not to be higher than 20°C. After the dripping is completed, the temperature is lowered to 0°C, and the reaction is stirred for 3 h, and a large amount of yellow solid is precipitated. The reaction solution is filtered, and the filter cake is washed with water (50 mL×2) and anhydrous ethanol (25 mL) in turn, and dried to obtain diacerein 2.

[0082] The synthetic route is as follows:

[0083]

[0084] Furthermore, the R 2 OH is methanol or menthol or borneol;

[0085] The catalyst A is concentrated sulfuric acid. When the etherification reaction is performed in step S1 to generate the intermediate 4-1, concentrated sulfuric acid is added. The hydrogen protons of the concentrated sulfuric acid combine with the hemiacetal hydroxyl group on the furanone 4-2 to form a pseudo-salt, which is then dehydrated to obtain a carbon cation, which is then reacted with R 2 OH attack to obtain the etherified product 4-1;

[0086] The acid-binding agent B is triethylamine. When compound 4 is generated in step S1, hydrohalic acid is generated. Triethylamine is added as an acid-binding agent to neutralize the hydrohalic acid to promote the reaction. When rhein-piperazine-furanone hybrid 5 is generated in step S2, hydrochloric acid is generated. Triethylamine is added as an acid-binding agent to neutralize the hydrochloric acid to promote the reaction.

[0087] The catalyst C is N,N-dimethylformamide, and when the intermediate 3 is generated in step S2, the reaction time is 2-3h, and the concentration of rhein or diacerein added to the polar aprotic solvent is 0.025-0.1M, so as to avoid the reaction time being too long, the amount of solvent in the reaction being too small, and the condensation reaction between molecules, which affects the generation of the intermediate 3; and when the rhein-piperazine-furanone hybrid is generated in step S2, the reaction temperature is 0-25°C, the reaction time is 1.5-3h, and the dropping rate of the acyl chloride solution is 0.5-1.5mL / min. Since the intermediate 3 is very active, the reaction temperature, time and the dropping rate of the acyl chloride solution are controlled during the reaction, so that the intermediate 3 reacts mildly with the compound 4, so as to reduce the condensation of the acyl chloride 3 with the hydroxyl groups at positions 1 and 8 of the rhein to generate esters, reduce the number of by-products, and improve the yield of the target product.

[0088] The aprotic solvent is 1,2-dichloroethane or cyclohexane;

[0089] The polar aprotic solvent is dichloromethane.

[0090] A pharmaceutically acceptable salt of a rhein-piperazine-furanone hybrid as described above.

[0091] A pharmaceutical composition of the rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof as described above.

[0092] The pharmaceutical preparation of the rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof as described above further comprises at least one pharmaceutically acceptable excipient or carrier.

[0093] The pharmaceutical preparation of the above-mentioned rhein-piperazine-furanone hybrid or a pharmaceutically acceptable salt thereof is a tablet, a capsule, a granule or an injection.

[0094] Use of the above-mentioned rhein-piperazine-furanone hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation in the preparation of anticancer drugs.

[0095] Use of the above-mentioned rhein-piperazine-furanone hybrid or its pharmaceutically acceptable salt or its pharmaceutical composition or its pharmaceutical preparation in the preparation of drugs against lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer.

[0096] For ease of understanding, the present invention is further illustrated by the following examples:

[0097] Embodiment 1, embodiment 2:

[0098] Add mucochloric acid or mucobromic acid (30 mmol), methanol (1 mL every 8 hours), and 1,2-dichloroethane (90 mL) into a flask and stir for 10 minutes, then drop 0.5 mL of concentrated sulfuric acid and continue stirring for 20 minutes. Then install a water separator, heat and reflux for 36 hours, stop heating, cool the mixture to room temperature, wash with saturated sodium carbonate (2×40 mL) and saturated brine (40 mL), dry the organic phase with anhydrous Na2SO4, remove the solvent by distillation under reduced pressure, and separate the residue by silica gel column chromatography to obtain 4-1e and 4-1h;

[0099] At room temperature, anhydrous piperazine (0.26 g, 3.0 mmol) was dissolved in 20 mL of dry dichloromethane, and then triethylamine (0.4 mL, 3.0 mmol) and compound 4-1e or 4-1h (2.0 mmol) were added in sequence. After the reaction was completed by TLC monitoring, the organic phase was washed with saturated sodium bicarbonate (2×30 mL) and saturated brine (30 mL) in sequence, and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure, and the residue was separated by silica gel column chromatography to obtain compound 4e or compound 4h. The yield of compound 4e was 65%, and the yield of compound 4h was 78%;

[0100] At room temperature, ultra-dry DMF (2-3 drops) was added dropwise to a solution of oxalyl chloride (4 mmol, 0.34 mL) in dichloromethane (40 mL). After stirring for 30 min, diacerein (1 mmol) was added. The temperature was raised to 30 °C and the reaction was continued for 3 h. The heating was stopped, the reaction solution was cooled to room temperature, and the solvent was removed by distillation under reduced pressure to obtain a crude acid chloride 3.

[0101] At 0°C, a solution of the crude acid chloride 3 in anhydrous dichloromethane (40 mL) was slowly dropped into a solution of compound 4e or 4h (1.2 mmol) and triethylamine (2 mmol, 0.3 mL) in anhydrous dichloromethane (30 mL). After reacting for about 2 h, the reaction was stopped and the reaction mixture was washed with saturated brine (20 mL×3). The organic phase was dried over anhydrous Na2SO4 and filtered. The solvent was removed by distillation under reduced pressure and separated by silica gel column chromatography to obtain rhein-piperazine-furanone hybrid 5e or 5h. The yield of rhein-piperazine-furanone hybrid 5e was 52%, and the yield of rhein-piperazine-furanone hybrid 5h was 63%.

[0102] Embodiment 3, Embodiment 4, Embodiment 5, Embodiment 6:

[0103] The method of Example 1 was used to replace only methanol with menthol or borneol, and the aprotic solvent 1,2-dichloroethane was replaced with cyclohexane. The crude product 4 was recrystallized with petroleum ether several times to obtain compound 4-1a or 4-1c or 4-1b or 4-1d, and then reacted with piperazine to obtain compound 4a or 4c or 4b or 4d. The yield of compound 4a was 87%, the yield of compound 4c was 88%, the yield of compound 4b was 78%, and the yield of compound 4d was 85%, 4a or 4c or 4b or 4d is further subjected to the subsequent reaction as in Example 1, and diacerein is replaced by rhein to obtain rhein-piperazine-furanone hybrid 5a or 5c or 5b or 5d, the yield of rhein-piperazine-furanone hybrid 5a is 61%, the yield of rhein-piperazine-furanone hybrid 5c is 76%, the yield of rhein-piperazine-furanone hybrid 5b is 86%, and the yield of rhein-piperazine-furanone hybrid 5d is 83%.

[0104] Embodiment 7, Embodiment 8, Embodiment 9, Embodiment 10:

[0105] By using the methods of Examples 3 to 6, rhein was replaced with diacerein to obtain rhein-piperazine-furanone hybrid 5f, 5i, 5g or 5j. The yield of rhein-piperazine-furanone hybrid 5f was 56%, the yield of rhein-piperazine-furanone hybrid 5i was 48%, the yield of rhein-piperazine-furanone hybrid 5g was 53%, and the yield of rhein-piperazine-furanone hybrid 5j was 69%.

[0106] The structural formulas of compounds 4-1e, 4-1h, 4-1a, 4-1c, 4-1b, and 4-1d are as follows:

[0107]

[0108] The structural formulas of compounds 4e, 4h, 4a, 4c, 4b and 4d are as follows:

[0109]

[0110] The structural formulas of rhein-piperazine-furanone hybrids 5a-5j are as follows:

[0111]

[0112] Please see Figures 1 to 30 ,pass 1 H NMR, 13 The products prepared in Examples 1 to 10 were characterized by C NMR and HRMS.

[0113] Hybrid 5a: (c 1.00, CH2Cl2);1 1H NMR (400 MHz, CDCl3) δ 11.95 (s, 1H), 11.79 (s, 1H), 7.68 (dd, J = 6.7, 1.5 Hz, 2H), 7.62 (t, J = 7.9 Hz, 1H), 7.25 (d, J = 1.6 Hz, 1H), 7.21 (dd, J = 8.3, 1.4 Hz, 1H), 5.81 (s, 1H), 3.99–3.46 (m, 9H), 2.15 (dt, J = 9.3, 3.2 Hz, 1H), 2.06 (pd, J = 6.6, 2.1 Hz, 1H), 1.60 (ddt, J = 15.1, 8.0, 3.3 Hz, 2H), 1.44–1.20 (m, 2H), 1.14–1.00 (m, 1H), 0.94 (dd, J = 12.7, 3.2 Hz, 1H), 0.90–0.83 (m, 6H), 0.83–0.74 (m, 1H), 0.70 (d, J = 6.9 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 192.2, 180.5, 167.7, 167.3, 162.5, 162.4, 154.9, 143.2, 137.7, 133.9, 132.9, 125.0, 122.5, 120.2, 117.7, 116.3, 115.3, 97.1, 88.9, 81.1, 48.0, 47.4, 42.3, 41.8, 33.7, 31.5, 25.1, 22.6, 22.2, 21.1, 15.7; IR (KBr) ν / cm -1 : 2961, 2926, 1759, 1631, 1454, 1288, 1263, 1205, 1158, 1118, 1091, 1051, 1016, 974, 799; HRMS (ESI) calcd for C 33 H 36 ClN2O8 [M + H]+ + 623.2155, found 623.2156.

[0114] Hybrid 5b: (c 1.00, CH2Cl2); 1HNMR(400MHz,CDCl3)δ11.94(s,1H),11.78(s,1H),7.67(dd,J=7.5,1.5Hz,2H),7.62(t,J=7.9Hz,1H),7.24(d,J=1.5Hz,1H),7.20(dd,J=8.2,1.4Hz,1H),5.73(d,J=3.1Hz,1H),4.14–3.30(m,9H),2.18(ddq,J=13.1,7.9,3.9Hz,1H),1.78(tt,J=11.1,4.2Hz,1H),1.72–1.56(m,2H),1.10(dtd,J=69.6,13.6,3.3Hz,3H),0.87–0.71(m,9H); 13 C NMR(101MHz,CDCl3)δ192.4,180.8,167.7,167.5,162.7,162.6,154.4,143.3,137.8,134.1,133.1,125.1,122.6,120.4,117.8,116.5,115.5,98.4,88.5,87.9,49.4,47.6,47.344.8,42.1,37.3,28.0,26.619.6,18.8,14.3;IR(KBr)ν / cm -1 :2956,1760,1628,1453,1378,1288,1266,1241,1206,1158,1126,1023,995,973,750;HRMS(ESI)calcd for C 33 H 34 ClN2O8[M+H] + 621.1998,found 621.2003.

[0115] Hybrid 5c: (c 1.00,CH2Cl2); 11H NMR (400 MHz, CDCl3) δ 12.04 (s, 1H), 11.87 (s, 1H), 7.76 (dd, J = 4.8, 1.5 Hz, 2H), 7.70 (t, J = 7.9 Hz, 1H), 7.32 (d, J = 1.5 Hz, 1H), 7.29 (dd, J = 8.3, 1.3 Hz, 1H), 5.89 (s, 1H), 4.06–3.51 (m, 9H), 2.27–2.19 (m, 1H), 2.13 (pd, J = 6.9, 2.2 Hz, 1H), 1.66 (ddt, J = 12.3, 8.4, 3.9 Hz, 2H), 1.47–1.26 (m, 2H), 1.13 (q, J = 11.9 Hz, 1H), 1.01 (dd, J = 12.8, 3.3 Hz, 1H), 0.95–0.91 (m, 6H), 0.88–0.81 (m, 1H), 0.77 (d, J = 6.9 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 192.2, 180.6, 168.1, 167.4, 162.6, 162.5, 157.6, 143.2, 137.7, 134.0, 133.0, 125.0, 122.5, 120.3, 117.8, 116.4, 115.4, 98.1, 81.0, 75.6, 48.0, 47.5, 42.3, 41.8, 33.8, 31.6, 25.2, 22.6, 22.2, 21.1, 15.7; IR (KBr) ν / cm -1 : 2956, 2925, 1755, 1626, 1453, 1377, 1287, 1265, 1240, 1206, 1157, 1117, 997, 967, 745; HRMS (ESI) calcd for C 33 H 36 BrN2O8 [M + H] + 667.1650, found 667.1650.

[0116] Hybrid 5d: (c 1.00, CH2Cl2); 11H NMR (400 MHz, CDCl3) δ 12.10 (s, 1H), 11.93 (s, 1H), 7.87–7.76 (m, 2H), 7.76–7.68 (m, 1H), 7.35–7.30 (m, 2H), 5.76 (d, J = 6.9 Hz, 1H), 4.20–3.46 (m, 9H), 2.32–2.18 (m, 1H), 1.83 (ddd, J = 14.0, 9.7, 4.2 Hz, 1H), 1.76–1.61 (m, 2H), 1.36–1.19 (m, 3H), 0.85 (d, J = 5.1 Hz, 9H); 13 13C NMR (101 MHz, CDCl3) δ 191.9, 180.2, 167.9, 167.1, 162.3, 162.1, 156.9, 143.2, 137.5, 133.6, 132.7, 124.8, 122.3, 120.0, 117.6, 116.0, 115.1, 99.1, 87.3, 74.5, 49.1, 47.3, 46.9, 44.6, 41.9, 37.0, 27.8, 26.4, 19.4, 18.6, 14.0; IR (KBr) ν / cm -1 : 2956, 1757, 1625, 1452, 1377, 1264, 1206, 1157, 1122, 1082, 1051, 1000, 968, 799, 743; HRMS (ESI) calcd for C 33 H 34 BrN2O8 [M + H] + 665.1493, found 665.1491.

[0117] Hybrid 5e: 1 1H NMR (400 MHz, CDCl3) δ 8.25–8.16 (m, 2H), 7.80 (t, J = 7.9 Hz, 1H), 7.49–7.35 (m, 2H), 5.68 (s, 1H), 3.78 (d, J = 63.5 Hz, 8H), 3.50 (s, 3H), 2.45 (d, J = 2.9 Hz, 6H); 13 13C NMR (101 MHz, CDCl3) δ 181.3, 180.2, 169.5, 169.4, 167.6, 167.2, 153.6, 150.6, 150.3, 140.7, 135.2, 134.8, 134.2, 130.8, 129.1, 126.6, 125.8, 125.6, 123.7, 97.6, 88.7, 55.5, 47.5, 42.2, 21.2; IR (KBr) ν / cm -1: 2956, 2921, 2850, 1765, 1676, 1624, 1594, 1470, 1445, 1324, 1256, 1196, 1143, 1011, 981; HRMS(ESI) calcd for C 28 H 27 ClN3O 10 [M + NH4] + 600.1379, found 600.1382.

[0118] Hybrid 5f: (c 1.00, CH2Cl2); 1 H NMR(400 MHz, CDCl3) δ 8.19(dd, J = 7.5, 1.4 Hz, 2H), 7.78(t, J = 7.9 Hz, 1H), 7.51–7.36(m, 2H), 5.80(s, 1H), 3.94–3.45(m, 9H), 2.43(d, J = 2.8 Hz, 6H), 2.18(d, J = 11.9 Hz, 1H), 2.07(pd, J = 7.0, 2.2 Hz, 1H), 1.64(dp, J = 15.1, 3.7 Hz, 2H), 1.33(dtd, J = 22.6, 11.9, 6.0 Hz, 2H), 1.10(q, J = 12.0 Hz, 1H), 0.97(dd, J = 12.8, 3.2 Hz, 1H), 0.90(dd, J = 6.8, 3.0 Hz, 6H), 0.85–0.79(m, 1H), 0.72(d, J = 6.9 Hz, 3H); 13 C NMR(101 MHz, CDCl3) δ 181.2, 180.2, 169.4, 169.2, 167.7, 167.0, 154.9, 150.5, 150.2, 140.6, 135.1, 134.7, 134.2, 130.7, 129.0, 126.5, 125.7, 125.5, 123.6, 97.2, 89.3, 81.2, 48.1, 47.4, 42.4, 42.0, 33.9, 31.7, 25.3, 22.7, 22.3, 21.2, 21.1, 15.8; IR(KBr) ν / cm -1 : 2959, 2925, 1770, 1677, 1631, 1429, 1368, 1324, 1289, 1259, 1195, 1119, 1016, 973, 802; HRMS(ESI) calcd for C 37 H 39 ClN2O 10 Na[M + Na] +729.2185, found 729.2141.

[0119] Hybrid 5g: (c 1.00, CH2Cl2); 1 H NMR (400MHz, CDCl3) δ8.26–8.11(m,2H),7.78(t,J=7.9Hz,1H),7.51–7.39(m,2H),5.71(d,J=7.6Hz,1H),4.16–3.50(m,9H),2.43 (d,J=3.1Hz,6H),2.23(ddt,J=17.7,8.8,3.7Hz,1H),1.94–1.74(m,1H),1.74–1.59(m,2H),1.37–0.93(m,3H),0.87–0.73(m,9H); 13 C NMR (101MHz, CDCl3) δ181.1,180.1,169.4,169.2,167.8,167.0154.4,150.5,150.1,140.7,135.1,134.7,134.1,130.7,129.0,1 26.4,125.6,125.5,123.6,98.4,88.4,87.8,49.4,47.6,47.4,44.8,42.2,37.2,28.0,26.6,21.1,19.6,18.8,14.3; IR(KBr)ν / cm -1 :1770,1678,1633,1429,1368,1325,1289,1259,1196,1123,1096,1017,983,973,802; HRMS(ESI)calcd for C 37 H 41 ClN3O 10 [M+NH4] + 722.2475, found 722.2475.

[0120] Hybrid 5h: 1 H NMR(400MHz, CDCl3))δ8.30–8.08(m,2H),7.78(t,J=7.9Hz,1H),7.57–7.36 (m,2H),5.68(s,1H),4.00–3.51(m,8H),3.46(s,3H),2.42(d,J=3.5Hz,6H); 13CNMR(101MHz,CDCl3)δ181.2,180.2,169.5,169.3,168.1,167.0,156.3,150.5,150.1,140.7,135.2,134.7,134.1,130.7,129.0,126.4,125.6,125.4,123.6,98.5,74.7,55.2,47.5,42.1,21.1;IR(KBr)ν / cm -1 :2921,1767,1677,1633,1594,1471,1431,1368,1325,1289,1257,1197,1122,1017,984;HRMS(ESI)calcd for C 28 H 27 BrN3O 10 [M+NH4] + 644.0874,found 644.0876.

[0121] Hybrid 5i: (c 1.00,CH2Cl2); 1 H NMR(400MHz,CDCl3)δ8.20(dd,J = 8.0,1.4Hz,2H),7.79(t,J = 7.9Hz,1H),7.56–7.38(m,2H),5.83(s,1H),3.95–3.50(m,9H),2.44(d,J = 2.8Hz,6H),2.19(dd,J = 12.4,4.4Hz,1H),2.08(pd,J = 7.1,2.2Hz,1H),1.64(ddt,J = 14.8,8.0,3.8Hz,2H),1.47–1.22(m,2H),1.11(q,J = 11.9Hz,1H),0.98(dd,J = 12.8,3.3Hz,1H),0.91(dd,J = 6.8,3.7Hz,6H),0.88–0.76(m,1H),0.73(d,J = 6.9Hz,3H); 1313C NMR (101 MHz, CDCl3) δ 181.2, 180.2, 169.5, 169.3, 168.2, 167.1, 157.6, 150.6, 150.2, 140.6, 135.2, 134.8, 134.2, 130.8, 129.1, 126.6, 125.7, 125.5, 123.7, 98.2, 81.2, 76.0, 48.2, 47.6, 42.4, 42.0, 33.9, 31.7, 25.3, 22.7, 22.3, 21.2, 21.1, 15.8; IR (KBr) ν / cm -1 : 2959, 2924, 1768, 1677, 1623, 1594, 1445, 1368, 1324, 1258, 1195, 1116, 1014, 966, 801; HRMS (ESI) calcd for C 37 H 43 BrN3O 10 [M + NH4] + 768.2126, found 768.2124.

[0122] Hybrid 5j: (c 1.00, CH2Cl2); 1 1H NMR (400 MHz, CDCl3) δ 8.25–8.14 (m, 2H), 7.79 (t, J = 7.9 Hz, 1H), 7.50–7.37 (m, 2H), 5.74 (d, J = 7.8 Hz, 1H), 4.16–3.53 (m, 9H), 2.43 (d, J = 3.2 Hz, 6H), 2.24 (ddd, J = 18.0, 9.2, 4.1 Hz, 1H), 1.81 (ddd, J = 14.1, 9.3, 4.4 Hz, 1H), 1.73–1.55 (m, 2H), 1.33–0.95 (m, 3H), 0.87–0.76 (m, 9H); 13 13C NMR (101 MHz, CDCl3) δ 181.2, 180.2, 169.4, 169.2, 168.2, 167.1, 157.0, 150.5, 150.2, 140.7, 135.2, 134.7, 134.2, 130.7, 129.0, 126.5, 125.7, 125.5, 123.6, 99.4, 87.8, 75.2, 49.5, 47.6, 47.5, 44.8, 42.2, 37.3, 28.0, 26.7, 21.1, 19.6, 18.8, 14.3; IR (KBr) ν / cm -1:2955,1767,1677,1624,1594,1432,1368,1325,1287,1258,1195,1122,1016,984,967; HRMS(ESI)calcd forC 37 H 41 BrN3O 10 [M+NH4] + 766.1970, found 766.1965.

[0123] The above characterization confirmed that the obtained product was the target product.

[0124] The following Table 1 is a list of compounds prepared in Examples 1 to 10:

[0125] Table 1

[0126]

[0127]

[0128] Comparative Example 1

[0129] Rhein 1 (1-1.5 mmol) reacts with oxalyl chloride in the presence of catalyst C and a polar aprotic solvent to obtain intermediate 3, which is then dripped into a dichloromethane solution (0.1-0.5 M) of piperazine (2-5 mmol) at 0°C using a constant pressure dropping funnel. After the dripping is complete, the mixture is returned to room temperature and the reaction is continued for 1-2 hours. After the reaction is stopped, the mixture is concentrated by spin drying and separated by column chromatography.

[0130] When rhein is first reacted with piperazine, since piperazine has a dinitrogen end, a disubstituted by-product will be generated. In addition, it was found during the experiment that the reaction system after the reaction is relatively complex, there are many by-products, and the newly generated rhein piperazine has poor solubility. A large amount of rhein piperazine cannot be obtained by column chromatography separation, acid-base extraction and recrystallization, which cannot support our subsequent synthesis work. Therefore, it is necessary to prepare according to the preparation method of the present invention, otherwise the target product cannot be obtained.

[0131] 1. The biological activity of 5a-5j prepared in the examples was tested:

[0132] 1) The biological activity of the prepared rhein-piperazine-furanone hybrids 5a-5j was studied to investigate the cytotoxicity and selectivity of the above compounds to human lung cancer cells A549 and human normal lung cells WI-38, wherein rhein (Rhein) and cytarabine (CAR) were used as control samples;

[0133] Experimental method: A549 cells and WI-38 cells in logarithmic growth phase with good growth status were selected and cultured at 4×10 3 100 μL of cells were inoculated into a 96-well culture plate and incubated overnight at 37°C (100 μL of sterile PBS was added to the wells around the cell wells);

[0134] After the cells adhered to the wall and grew well for 24 hours, the old culture medium was absorbed, and 10 μL of different concentrations of hybrid 5, Rhein and CAR (the concentrations were 1, 10, 50, 100, and 300 μg / mL, respectively; the concentration of Rhein in A549 cells was 1, 20, 100, 300, and 500 μg / mL) were added to each culture well of A549 and WI-38 cells. Three parallel replicate wells were set for each concentration, and blank control wells with equal volumes of dimethyl sulfoxide (DMSO) solvent and drug-free culture medium were set, and the cells were cultured in an incubator at 37°C and 5% CO2.

[0135] After culturing the test substance for 48 hours, the supernatant was discarded and the cell morphology was observed using an inverted microscope; then, 10 μL (2 mg / mL in PBS) MTT was added to each well, and the culture was continued for 4 hours. The cells were centrifuged at 1000 r / min for 10 minutes, the culture supernatant in the well was aspirated, 150 μL dimethyl sulfoxide was added to each well, and the cells were shaken for 10 minutes to fully dissolve the blue-purple crystals. The absorbance value (A) of each well sample was measured at a wavelength of 568 nm using an enzyme marker, and the average value was taken.

[0136] Cell inhibition rate = [A568 (negative control) - A568 (administration)] / A568 (negative control) × 100%.

[0137] The results are as follows Fig.31 As shown in Table 2:

[0138] Table 2

[0139]

[0140] A549 = human lung cancer cell line; WI-38 = human normal lung cell line; CAR = cytarabine; IC 50 The value represents the compound concentration (μM) required to inhibit tumor and normal cell proliferation by 50%, and is calculated using the concentrations measured three times. SI: To determine the safe range of drug action, the larger the index, the safer it is. Its value is the IC of normal cells 50 IC of tumor cells 50 .

[0141] from Fig.31 It can be clearly seen from Table 2 that all target hybrids 5a–j (IC 50=5.49–116.78 μM) were more cytotoxic to A549 cells than the parent Rhein (IC 50 =265.59 μM) and positive control CAR (IC 50 =202.57 μM). Structure-activity relationship (SAR) analysis showed that: R 2 When it is methyl (5e, IC 50 =5.74μM; 5h, IC 50 =5.49μM) was the most toxic to A549 cells, about 48 times that of the parent Rhein and 37 times that of the positive control CAR. More importantly, the selectivity (WI-38 / A549) of the hybrids 5e (SI≈12) and 5h (SI≈7) was also higher, better than that of the parent Rhein (SI≈1) and the positive control CAR (SI≈2), and therefore had good practical application value. 1 The hydrogen hybrids (5a–d, IC 50 =151.85–296.00 μM) is less toxic to normal cells than R 1 Hybrids with acetyl groups (5e–j, IC 50 =24.85-69.28 μM), which provides a reference for reducing the toxicity of rhein derivatives to normal cells.

[0142] Moreover, compared with the hybrid prepared in the patent document with application number 202210453596.5 mentioned in the background technology, the toxicity of the hybrid 5 prepared in the present invention to cancer cells is significantly improved, and the selectivity is better, especially the hybrids 5e and 5h are the most significant.

[0143] 2) In view of the toxicity of hybrid 5e to A549 cells (IC 50 =5.74 μM) and the highest selectivity (WI-38 / A549, SI≈12). We further tested the toxicity of hybrid 5e to 10 human cancer cells and 4 corresponding human normal cells. The experimental method was similar to that of A549 and WI-38 cells. The concentrations of hybrid 5e were 0.2, 5, 20, 50, and 100 μg / mL. The test results are shown in Table 3.

[0144] Table 3

[0145]

[0146]

[0147] A549 = human lung cancer cell line; Calu-1 = human lung adenocarcinoma cell line; PC-9 = human lung cancer cell line; H460 = human lung cancer cell line; SK-BR-3 = human breast cancer cell line; 786-O = human kidney cancer cell line; SK-OV-3 = human ovarian cancer cell line; Huh-7 = human liver cancer cell line; HCT116 = human colon cancer cell line; Hela = human cervical cancer cell line; WI-38 = human lung normal cell line; MCF-10A = human normal breast epithelial cell line; HEK-293T = human embryonic kidney cell line; KGN = human ovarian cell line.

[0148] The experimental results in Table 3 show that the hybrid 5e has strong toxicity to the above 10 human cancer cells, but relatively less toxicity to the corresponding normal human cells. The toxicity (IC 50 =4.35–15.39 μM) was superior to 6 other human cancer cells (SK-BR-3, 786-O, SK-OV-3, Huh-7; HCT116, Hela; IC 50 =13.77–60.45 μM) showed obvious specificity. In particular, the toxicity of hybrid 5e to H460 cells (IC 50 =4.35μM) and had the highest selectivity (WI-38 / H460, SI≈16).

[0149] 3) The apoptosis-inducing effect of hybrid 5e on H460 cells was further studied by flow cytometry analysis. The experimental method was: H460 cells (1×10 6 in 100 μL) were seeded in 6-well plates in triplicate and incubated at 37°C for 24 h. The medium was replaced with fresh medium containing hybrid 5e at the desired concentration (0.5, 5, 50 μg / mL). An equal amount of DMSO negative control well was set up. The cells were incubated at 37°C for 48 h, and 0.25% trypsin without ethylenediaminetetraacetic acid (EDTA) was added to each well. After trypsin treatment, the treated cells were stained with the AnnexinV-FITC / PI apoptosis detection kit according to the manufacturer's instructions. After incubation in the dark at room temperature for 5–15 minutes, apoptotic cells were immediately analyzed using a CytoFLEX flow cytometer, and the detection results were shown as follows. Fig.32 shown.

[0150] like Fig.32 As shown in A, with the increase of drug concentration, the early apoptosis rate and late apoptosis rate of H460 cells gradually increased; Fig.32As shown in B, when the hybrid 5e was treated with 0.5, 5 and 50 μg / mL of H460 cells for 48 h, the total apoptosis rates were 9.38%, 37.72% and 56.57%, respectively, while the control group was only 6.85%. The above results indicate that the hybrid 5e induces apoptosis of H460 cells in a concentration-dependent manner.

[0151] What is disclosed above is only a preferred embodiment of the present invention, which certainly cannot be used to limit the scope of rights of the present invention. A person skilled in the art can understand that all or part of the processes of the above embodiments and equivalent changes made according to the claims of the present invention still fall within the scope of the invention.

Claims

1. A rhein-piperazine-furanone hybrid, characterized in that: The general structural formula of the rhein-piperazine-furanone hybrid is as follows: in, X is a chlorine atom or a bromine atom, R 1 is selected from hydrogen or acetyl, R 2 Selected from methyl, menthyl or bornyl.

2. A method for preparing a rhein-piperazine-furanone hybrid, comprising preparing the rhein-piperazine-furanone hybrid as claimed in claim 1, characterized in that: The steps include: S1: Mix mucochloric acid or mucobromic acid with R 2 OH undergoes etherification reaction in catalyst A and aprotic solvent to obtain intermediate 4-1, and intermediate 4-1 undergoes nucleophilic substitution reaction with piperazine in acid-binding agent B and polar aprotic solvent to obtain compound 4; S2: Rhein, 1 or diacerein, 2 is subjected to chlorination reaction with oxalyl chloride in a catalyst C and a polar aprotic solvent to obtain an acyl chloride intermediate 3; intermediate 3 is subjected to N-acylation reaction with compound 4 in an acid-binding agent B and a polar aprotic solvent to obtain rhein-piperazine-furanone hybrid 5; The R 2 OH is methanol or menthol or borneol; The catalyst A is concentrated sulfuric acid; The acid binding agent B is triethylamine; The catalyst C is N,N-dimethylformamide; The aprotic solvent is 1,2-dichloroethane or cyclohexane; The polar aprotic solvent is dichloromethane; The synthetic route is as follows:

3. A pharmaceutically acceptable salt of the rhein-piperazine-furanone hybrid according to claim 1; Or a pharmaceutically acceptable salt of the rhein-piperazine-furanone hybrid prepared by the method for preparing the rhein-piperazine-furanone hybrid according to claim 2.

4. A pharmaceutical composition containing the rhein-piperazine-furanone hybrid as claimed in claim 1; or a pharmaceutical composition containing the rhein-piperazine-furanone hybrid prepared by the method for preparing the rhein-piperazine-furanone hybrid according to claim 2; Or a pharmaceutical composition containing a pharmaceutically acceptable salt of the rhein-piperazine-furanone hybrid according to claim 3.

5. A pharmaceutical preparation containing the rhein-piperazine-furanone hybrid as claimed in claim 1; or a pharmaceutical preparation containing the rhein-piperazine-furanone hybrid prepared by the method for preparing the rhein-piperazine-furanone hybrid according to claim 2; Or a pharmaceutical preparation containing a pharmaceutically acceptable salt of the rhein-piperazine-furanone hybrid according to claim 3, characterized in that: It also includes at least one pharmaceutically acceptable excipient or carrier.

6. The pharmaceutical preparation according to claim 5, characterized in that The pharmaceutical preparation is a tablet, a capsule, a granule or an injection.

7. Use of the rhein-piperazine-furanone hybrid according to claim 1 in the preparation of drugs for treating lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer; Or use of the rhein-piperazine-furanone hybrid prepared by the method for preparing the rhein-piperazine-furanone hybrid according to claim 2 in the preparation of drugs for treating lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer; or use of a pharmaceutically acceptable salt of the rhein-piperazine-furanone hybrid according to claim 3 in the preparation of a drug for treating lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer; or use of the pharmaceutical composition according to claim 4 in the preparation of a drug for treating lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer; Or use of the pharmaceutical preparation according to claim 5 or 6 in the preparation of drugs for treating lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, colon cancer or cervical cancer.

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