Rhein-piperazine-sulfonamide hybrids, processes for their preparation and use

Abstract

The present application discloses a rhein-piperazine-sulfamide hybrid and a preparation method and application thereof. The rhein-piperazine-sulfamide hybrid is a compound shown in formula (I) or a pharmaceutically acceptable salt thereof: wherein R 1 is selected from hydrogen or acetyl; R 2 is selected from aryl or heteroaryl which is unsubstituted or substituted with alkyl, alkoxy, halogen or nitro. The present application combines rhein or diacerein as a starting material, piperazine as a linking group, and a sulfamide derivative into one molecule, to obtain an anticancer candidate drug which is more active, more selective and safer than the parent rhein.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a rhein-piperazine-sulfonamide hybrid, its preparation method, and its application. Background Technology

[0002] Cancer is a leading cause of death, causing approximately 9 million deaths annually, making it the second leading cause of death worldwide after heart disease. Anticancer drugs are crucial in cancer treatment, and currently, over 100 drugs have been approved for cancer treatment. However, the emergence of drug resistance has led to a decrease in the effectiveness of clinically used chemotherapy agents. Therefore, developing novel, effective, and safe anticancer drugs remains an important task in medicinal chemistry research. Summary of the Invention

[0003] Rhein is a natural anthraquinone drug widely found in herbs such as rhubarb and Polygonum multiflorum. Its poor bioavailability and gastrointestinal irritation limit its clinical application. Anthraquinone derivatives, such as diacerein, valrubicin, and idarubicin, have been used to treat inflammation, solid tumors, and hematological malignancies.

[0004] Piperazine is an important N-heterocyclic compound widely found in bioactive compounds. Sulfonamide derivatives are a class of compounds with diverse biological activities and promising applications; many have been used in cancer therapy, such as Indisulam, Belinostat, and Amsacrine. Piperazine-sulfonamides can inhibit tumor growth and are attractive scaffolds due to their simple synthetic pathways, low toxicity, and metabolic stability. Introducing piperazine-sulfonamide fragments into anticancer drugs can lead to increased binding affinity and binding strength.

[0005]

[0006] Therefore, this invention uses Rhein or Diacerein as the starting material and piperazine as the linking group, combining it with a sulfonamide derivative into a single molecule, with the aim of obtaining an anticancer drug candidate that is more active, more selective, and safer than the parent Rhein.

[0007] To achieve the above objectives, the embodiments of the present invention provide the following technical solutions:

[0008] According to a first aspect of the present invention, the present invention provides a rhein-piperazine-sulfonamide hybrid, said rhein-piperazine-sulfonamide hybrid being a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0009]

[0010] Among them, R 1 Selected from hydrogen or acetyl groups;

[0011] R 2 Selected from unsubstituted aryl or heteroaryl groups or those substituted with alkyl, alkoxy, halogen or nitro groups.

[0012] Furthermore, the R 2 Selected from one of the following groups:

[0013]

[0014] According to a second aspect of the present invention, the present invention provides a method for preparing the rhein-piperazine-sulfonamide hybrid as described above, characterized in that the synthetic route is as follows:

[0015]

[0016] The preparation method includes:

[0017] Rhein 1 or diacerein 2 reacts with oxalyl chloride in the presence of a catalyst and a polar aprotic solvent to give intermediate 3.

[0018] Intermediate 3 and piperazine-sulfonamide derivative 4 were subjected to N-acylation in the presence of an acid-binding agent and a polar aprotic solvent to give compound 5, namely the compound shown in formula (I);

[0019] Among them, R in piperazine-sulfonamide derivative 4 2 The definition is as described above.

[0020] Furthermore, the catalyst is N,N-dimethylformamide; the polar aprotic solvent is dichloromethane; and the acid-binding agent is triethylamine.

[0021] According to a third aspect of the present invention, the present invention provides a pharmaceutical composition comprising the rhein-piperazine-sulfonamide hybrid as described above.

[0022] According to a fourth aspect of the present invention, the present invention provides a pharmaceutical formulation comprising the rhein-piperazine-sulfonamide hybrid as described above, and at least one pharmaceutically acceptable excipient or carrier.

[0023] Furthermore, the formulation is a tablet, capsule, granule, or injection.

[0024] According to a fifth aspect of the present invention, the present invention provides the use of the rhein-piperazine-sulfonamide hybrid, the pharmaceutical composition, or the pharmaceutical preparation described above in the preparation of an anticancer drug.

[0025] Furthermore, the cancer in question is human lung cancer, human liver cancer, or human breast cancer.

[0026] The embodiments of the present invention have the following advantages:

[0027] This invention synthesizes a series of novel rhein-piperazine-sulfonamide hybrids 5 by introducing a sulfonamide scaffold into rhein using a simple and efficient continuous reaction. The in vitro cytotoxicity of hybrids 5 against human hepatocellular carcinoma cells (HepG2), human breast cancer cells (MCF-7), human lung cancer cells (A549), and normal human hepatocytes (LO2) was evaluated using the MTT assay. The results showed that hybrids 5 exhibited superior cytotoxicity against HepG2 hepatocellular carcinoma cells, MCF-7 breast cancer cells, and A549 lung cancer cells compared to the parent Rhein, with particularly strong inhibitory effects against A549 lung cancer cells. Structure-activity relationship analysis indicated that the introduction of the sulfonamide scaffold significantly enhanced the cytotoxicity of the parent Rhein. Hybrids 5j (IC 50 =7.00 μM) was the most toxic to A549 lung cancer cells, approximately equal to that of the parent Rhein (IC50). 50 =142.23 μM) 20 times more selective than the parent compound Rhein (SI≈14, LO2 / A549) and the clinical anticancer drug doxorubicin (DOX, SI≈0). Attached Figure Description

[0028] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0029] Figure 1 The rhein-piperazine-sulfonamide hybrid 5a provided by this invention 1 H NMR spectrum;

[0030] Figure 2 The rhein-piperazine-sulfonamide hybrid 5a provided by this invention 13 C NMR spectrum;

[0031] Figure 3 The HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5a provided by the present invention;

[0032] Figure 4 The rhein-piperazine-sulfonamide hybrid 5b provided by this invention 1 H NMR spectrum;

[0033] Figure 5The rhein-piperazine-sulfonamide hybrid 5b provided by this invention 13 C NMR spectrum;

[0034] Figure 6 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5b provided by the present invention;

[0035] Figure 7 The rhein-piperazine-sulfonamide hybrid 5c provided by this invention 1 H NMR spectrum;

[0036] Figure 8 The rhein-piperazine-sulfonamide hybrid 5c provided by this invention 13 C NMR spectrum;

[0037] Figure 9 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5c provided by the present invention;

[0038] Figure 10 The rhein-piperazine-sulfonamide hybrid 5d provided by this invention 1 H NMR spectrum;

[0039] Figure 11 The rhein-piperazine-sulfonamide hybrid 5d provided by this invention 13 C NMR spectrum;

[0040] Figure 12 The rhein-piperazine-sulfonamide hybrid 5d provided by this invention 19 F NMR spectrum;

[0041] Figure 13 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5d provided by the present invention;

[0042] Figure 14 The rhein-piperazine-sulfonamide hybrid 5e provided by this invention 1 H NMR spectrum;

[0043] Figure 15 The rhein-piperazine-sulfonamide hybrid 5e provided by this invention 13 C NMR spectrum;

[0044] Figure 16 The HRMS mass spectrum of the rhein-piperazine-sulfonamide hybrid 5e provided by this invention;

[0045] Figure 17 The rhein-piperazine-sulfonamide hybrid 5f provided by this invention 1 H NMR spectrum;

[0046] Figure 18 The rhein-piperazine-sulfonamide hybrid 5f provided by this invention 13 C NMR spectrum;

[0047] Figure 19 The HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5f provided by the present invention;

[0048] Figure 20 The present invention provides 5g of rhein-piperazine-sulfonamide hybrid. 1 H NMR spectrum;

[0049] Figure 21 The present invention provides 5g of rhein-piperazine-sulfonamide hybrid. 13 C NMR spectrum;

[0050] Figure 22 HRMS mass spectrum of 5g of rhein-piperazine-sulfonamide hybrid provided by the present invention;

[0051] Figure 23 The rhein-piperazine-sulfonamide hybrid provided by this invention has a 5h [result]. 1 H NMR spectrum;

[0052] Figure 24 The rhein-piperazine-sulfonamide hybrid provided by this invention has a 5h [result]. 13 C NMR spectrum;

[0053] Figure 25 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid for 5 h provided by the present invention;

[0054] Figure 26 The rhein-piperazine-sulfonamide hybrid 5i provided by this invention 1 H NMR spectrum;

[0055] Figure 27 The rhein-piperazine-sulfonamide hybrid 5i provided by this invention 13 C NMR spectrum;

[0056] Figure 28 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5i provided by the present invention;

[0057] Figure 29 The rhein-piperazine-sulfonamide hybrid 5j provided by this invention 1 H NMR spectrum;

[0058] Figure 30 The rhein-piperazine-sulfonamide hybrid 5j provided by this invention13 C NMR spectrum;

[0059] Figure 31 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5j provided by the present invention;

[0060] Figure 32 The rhein-piperazine-sulfonamide hybrid 5k provided by this invention 1 H NMR spectrum;

[0061] Figure 33 The rhein-piperazine-sulfonamide hybrid 5k provided by this invention 13 C NMR spectrum;

[0062] Figure 34 The rhein-piperazine-sulfonamide hybrid 5k provided by this invention 19 F NMR spectrum;

[0063] Figure 35 HRMS mass spectrum of 5k rhein-piperazine-sulfonamide hybrid provided by the present invention;

[0064] Figure 36 The rhein-piperazine-sulfonamide hybrid 5l provided by the present invention 1 H NMR spectrum;

[0065] Figure 37 The rhein-piperazine-sulfonamide hybrid 5l provided by the present invention 13 C NMR spectrum;

[0066] Figure 38 HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5l provided by the present invention;

[0067] Figure 39 The rhein-piperazine-sulfonamide hybrid 5m provided by this invention 1 H NMR spectrum;

[0068] Figure 40 The rhein-piperazine-sulfonamide hybrid 5m provided by this invention 13 C NMR spectrum;

[0069] Figure 41 The HRMS mass spectrum of the rhein-piperazine-sulfonamide hybrid at 5m provided by this invention;

[0070] Figure 42 The rhein-piperazine-sulfonamide hybrid 5n provided by this invention 1 H NMR spectrum;

[0071] Figure 43The rhein-piperazine-sulfonamide hybrid 5n provided by this invention 13 C NMR spectrum;

[0072] Figure 44 The HRMS mass spectrum of rhein-piperazine-sulfonamide hybrid 5n provided by this invention;

[0073] Figure 45 The inhibitory effects of different concentrations of hybrid 5 provided by this invention on A549 and LO2 cell lines. Detailed Implementation

[0074] The following specific embodiments illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0075] All required reagents were purchased from Adamas, Aldrich, Energy Chemical, and J&K Chemical, and should be used directly unless otherwise specified. Dichloromethane at P4O 10 Dry on silica gel GF. 254 Thin-layer chromatography (TLC) was performed on the plate (Qingdao Ocean Chemical Co., Ltd.). Infrared spectra were recorded using KBr pellets on an FTIR-8400S spectrometer. Results were obtained using a Bruker Avance III 400MHz spectrometer and a Bruker Avance III 500MHz spectrometer. 1 H NMR, 13 C NMR and 19 F NMR spectrum. Chemical shift (δ) of tetramethylsilane in the ppm range was used as an internal standard. Residual solvent signal was used as... 1 H NMR and 13 Reference for CNMR spectrum (CDCl3:δ) H =7.26ppm, δ C =77.16ppm; DMSO-d6:δ H =2.50ppm, δ C =39.52ppm). High-resolution mass spectrometry (HRMS) was tested on a Thermo Fisher LTQ Orbitrap XL.

[0076] The synthetic route and preparation method of piperazine-sulfonamide derivative 4 are as follows:

[0077]

[0078] Piperazine (1.72 g, 20 mmol) was dissolved in 100 mL of dichloromethane at 0 °C. The mixture was stirred until all the piperazine was dissolved. Sulfonyl chloride (5 mmol) was added, and the reaction was continued for 0.5–1 hour before the reaction was stopped. The reaction solution was washed with saturated NaHCO3 solution (50 mL × 2) and saturated saline solution (50 mL × 2), respectively. The solution was concentrated by vacuum distillation, and the residue was purified by silica gel column chromatography to obtain compound 4.

[0079] Compound 4a, a colorless oily liquid, yield 96%. 1 H NMR (400MHz, DMSO-d6) δ7.72(d,J=7.2Hz,3H),7.68–7.63(m,2H),3.07–2.87(m,1H),2.81–2.74(m,4H),2.73–2.67(m,4H). 13 C NMR (101MHz, DMSO-d6) δ134.85,133.25,129.44,127.67,46.83,44.74.

[0080] Compound 4b, a colorless oily liquid, in 96% yield. 1 H NMR (400MHz, DMSO-d6) δ7.60(d,J=8.2Hz,2H),7.45(d,J=8.0Hz,2H),2.77–2.72(m,4H),2.71–2.67(m,4H),2.41(s,3H). 13 C NMR (101MHz, DMSO-d6) δ143.63,131.92,129.85,127.73,46.83,44.74,21.07.

[0081] Compound 4c, white solid, yield 87%. 1 H NMR (400MHz, DMSO-d6) δ7.66(d,J=8.9Hz,2H),7.16(d,J=9.0Hz,2H),3.85(s,3H),2.83–2.78(m,4H),2.78–2.74(m,4H). 13 C NMR (101MHz, DMSO-d6) δ162.86,129.89,126.34,114.56,55.81,46.25,44.37.

[0082] Compound 4d, white solid, yield 83%. 1H NMR (400MHz, DMSO-d6) δ7.71(tt,J=9.2,2.3Hz,1H),7.51–7.43(m,2H),2.90–2.82(m,4H),2.75–2.68(m,4H). 13 C NMR (101MHz, DMSO-d6) δ162.51(d,J=252.5Hz), 162.39(d,J=253.5Hz), 138.54(t,J=8. 1Hz), 111.40 (d, J = 28.3Hz), 111.40 (d, J = 11.1Hz), 109.07 (t, J = 25.3Hz), 46.86, 44.74. 19 F NMR(376MHz,DMSO-d6)δ-106.11.

[0083] Compound 4e, white solid, yield 62%. 1 H NMR (400MHz, DMSO-d6) δ7.70(d,J=2.8Hz,1H),7.67(s,1H),7.30(d,J=8.6Hz,1H ),3.90(s,3H),3.01(t,J=4.8Hz,4H),2.68(t,J=4.9Hz,4H),2.48–2.16(m,1H). 13 C NMR (101MHz, DMSO-d6) δ155.59,134.35,129.92,127.03,123.80,115.23,56.46,46.66,45.32.

[0084] Compound 4f, white solid, yield 69%. 1 H NMR (400MHz, DMSO-d6) δ8.45(d,J=8.6Hz,2H),7.99(d,J=8.6Hz,2H),2.85(t,J=4.9Hz,4H),2.72(t,J=4.9Hz,4H),2.32(s,1H). 13 C NMR (101MHz, DMSO-d6) δ150.12,140.72,129.20,124.70,46.78,44.79.

[0085] Compound 4g, pale yellow solid, yield 69%. 1H NMR (400MHz, DMSO-d6) δ7.68(tt,J=9.2,2.5Hz,1H),7.46(d,J=4.0Hz,1H),2.85(t,J=4.9Hz,4H),2.72(t,J=4.9Hz,4H),2.44–2.11(m,1H). 13 C NMR (101MHz, DMSO-d6) δ135.78,133.69,132.01,119.34,46.65,44.52.

[0086] The synthetic route and general preparation method of the target product, rhein-piperazine-sulfonamide hybrid 5, are as follows:

[0087]

[0088] Synthetic reagents and conditions: a) Acetic anhydride, concentrated sulfuric acid, -10℃ to 35℃, 3 hours; b) Dichloromethane, N,N-dimethylformamide, oxaloyl chloride, 30℃, 1 hour; c) Dichloromethane, triethylamine, 0℃, 1 hour.

[0089] Rhein 1 and diacerein 2 (prepared from rhein 1 via acetylation) reacted with oxalyl chloride to give acyl chloride 3, which then underwent N-acylation with compound 4 to yield rhein-piperazine-sulfonamide hybrid 5. The N-acylation reaction required controlled temperature at 0°C, and the rate of addition of acyl chloride 3 had to be strictly controlled; a slower addition rate resulted in a more homogeneous product. The reaction time should not be too long; it should be stopped after approximately 1–2 hours. The mixture was then concentrated by vacuum distillation, and the residue was purified by silica gel column chromatography to obtain the target compound 5.

[0090] Specifically, the preparation method of rhein-piperazine-sulfonamide hybrid 5 is as follows:

[0091] Preparation of diacerein 2: At -10℃, concentrated sulfuric acid (1.76 mL, 22 mmol) was slowly added dropwise to a solution of rhein 1 (2.84 g, 10 mmol) and acetic anhydride (3.06 g, 30 mmol), completing the addition in approximately 1 hour. The mixture was then heated to 35℃ and stirred for 3 hours. The reaction solution was then slowly added dropwise to 1.70 L of water at 0℃, with the temperature inside the flask controlled to not exceed 20℃. After the addition was complete, the mixture was cooled to 0℃ and stirred for 3 hours, resulting in the precipitation of a large amount of yellow solid. The reaction solution was filtered, and the filter cake was washed successively with water (100 mL × 2) and anhydrous ethanol (250 mL), and dried to obtain diacerein 2 (3.62 g).

[0092] Preparation of rhein-piperazine-sulfonamide hybrid 5: At room temperature, ultra-dry DMF (2-3 drops) and oxaloyl chloride (0.34 mL, 4 mmol) were added dropwise to a dichloromethane (50 mL) solution containing rhein 1 (284.22 mg, 1.0 mmol) or diacerein 2 (368.29 mg, 1.0 mmol). The temperature was raised to 30 °C, and the reaction was carried out for 1 h. Heating was stopped, and the reaction solution was cooled to room temperature and concentrated by vacuum distillation to obtain crude acyl chloride 3. At 0 °C, an anhydrous dichloromethane (50 mL) solution of the above crude acyl chloride 3 was slowly added dropwise to an anhydrous dichloromethane (20 mL) solution containing compound 4 (1.1 mmol) and triethylamine (0.21 mL, 1.5 mmol). The reaction was carried out for 1 h, and the reaction was stopped. The solution was directly concentrated by vacuum distillation, and the residue was separated and purified by silica gel column chromatography to obtain rhein-piperazine-sulfonamide hybrid 5.

[0093] Rhein-piperazine-sulfonamide hybrid 5a: reddish-brown solid, yield 57%, mp 180.8–181.6 °C; 1 HNMR(400MHz,DMSO-d6)δ11.87(s,2H),7.79(d,J=7.9Hz,1H),7.75(d,J=7.3Hz,3H),7.69(s,1H),7.68–7 .64(m,2H),7.56(s,1H),7.35(d,J=8.4Hz,2H),3.79–3.67(m,2H),3.49–3.39(m,2H),3.10–2.92(m,4H). 13 C NMR(101MHz,DMSO-d6)δ191.34,180.91,166.59,161.37,161.15,143.62,137.56,135.15,133.73,133.4 4,133.14,129.57,127.53,124.61,122.00,119.43,117.31,116.42,115.90,46.22,45.38;IR(KBr)ν / cm -1 :1629,1452,1350,1277,1268,1208,1168,1143,739,576; HRMS(ESI)calcd for C 25 H 21 N₂O₇S[M+H] + 493.1064, found 493.1065.

[0094] Rhein-piperazine-sulfonamide hybrid 5b: reddish-brown solid, yield 61%, mp 137.8–138.5 °C; 1HNMR(400MHz,DMSO-d6)δ11.85(s,2H),7.76(t,J=8.0Hz,1H),7.65–7.60(m,3H),7.55(s,1H),7.47(s,1H),7. 45(d,J=4.0Hz,1H),7.38–7.32(m,2H),3.78–3.70(m,2H),3.49–3.39(m,2H),3.05–2.90(m,4H),2.42(s,3H). 13 C NMR(101MHz,DMSO-d6)δ191.31,180.83,166.55,161.37,161.14,143.84,143.62,137.55,133.67,133.08,1 32.18,129.99,127.58,124.59,122.02,119.42,117.33,116.35,115.83,46.06,45.38,21.06;IR(KBr)ν / cm -1 :1630,1452,1349,1278,1267,1203,1165,752,725,548; HRMS(ESI)calcd for C 26 H 23 N₂O₇S[M+H] + 507.1220, found 507.1221.

[0095] Rhein-piperazine-sulfonamide hybrid 5c: brown solid, yield 64%, mp 198.7–199.5 °C; 1 H NMR (400MHz, DMSO-d6) δ11.90(s,1H),11.82(s,1H),7.77(t,J=7.9Hz,1H),7.68(s,1H),7.65(d,J=8.1Hz,2H),7.56(s, 1H),7.35(t,J=4.3Hz,2H),7.17(d,J=8.4Hz,2H),3.86(s,3H),3.79–3.67(m,2H),3.50–3.38(m,2H),3.05–2.88(m,4H). 13C NMR(101MHz,DMSO-d6)δ191.34,180.88,166.55,162.89,161.36,161.14,143.64,137.54,133.71,133.12,1 29.79,126.50,124.59,121.99,119.42,117.31,116.39,115.87,114.67,55.73,45.64,45.38; IR(KBr)ν / cm -1 :1630,1596,1453,1349,1266,1202,1186,1160,729,558; HRMS(ESI)calcd forC 26 H 22 N₂NaO₈S[M+Na] + 545.0989, found 545.0978.

[0096] Rhein-piperazine-sulfonamide hybrid 5d: brown solid, yield 66%, mp 187.6–188.4℃; 1 H NMR (400MHz, DMSO-d6) δ11.86(s,1H),11.76(s,1H),7.73(s,1H),7.70(d,J=9.2Hz,1H),7.59(d,J=7.5Hz,1H),7.55(s,1H ),7.49(s,1H),7.48(s,1H),7.33(s,1H),7.29(d,J=8.4Hz,1H),3.82–3.69(m,2H),3.51–3.39(m,2H),3.23–3.02(m,4H). 13 C NMR(101MHz,DMSO-d6)δ191.26,180.68,166.51,163.69(d,J=12.1Hz),16 1.35,161.25,161.14,143.60,138.70(t,J=8.1Hz),137.50,133.63,132.9 7,123.23(d,J=261.6Hz),118.37(d,J=205.0Hz),116.25,115.69,111.32 (d,J=28.3Hz),111.32(d,J=12.1Hz),109.19(t,J=26.3Hz),46.17,45.50; 19 F NMR(376MHz,DMSO-d6)δ-105.90; IR(KBr)ν / cm -1:1630,1606,1452,1441,1360,1296,1267,1201,1166,594; HRMS(ESI)calcd for C 25 H 19 F2N2O7S[M+H] + 529.0875, found 529.0874.

[0097] Rhein-piperazine-sulfonamide hybrid 5e: yellow solid, yield 67%, mp 236.2–237.1℃; 1 H NMR (500MHz, DMSO-d6) δ11.87(s,2H),7.78(t,J=8.0Hz,1H),7.71(dd,J=8.8,2.7Hz,1H),7.69–7.65(m,2H),7.61(s,1H),7.3 8(s,1H),7.36(d,J=9.0Hz,1H),7.32(d,J=8.9Hz,1H),3.92(s,3H),3.73–3.63(m,2H),3.44–3.35(m,2H),3.31–3.15(m,4H). 13 C NMR (126MHz, DMSO-d6) δ191.30,180.84,166.57,161.35,161.15,155.64,143.72,137.50,134.54,133.72,133.10,1 29.66,127.10,124.54,123.77,121.90,119.38,117.26,116.37,115.84,115.38,56.53,46.77,45.23.IR(KBr)ν / cm -1 :1730,1651,1542,1414,1306,1288,1276,1251,1106,781; HRMS(ESI)calcd for C 26 H 22 ClN2O8S[M+H] + 557.0780, found 557.0772.

[0098] Rhein-piperazine-sulfonamide hybrid 5f: yellow solid, yield 63%, mp 221.5–222.3℃; 1H NMR (500MHz, DMSO-d6) δ11.88(s,1H),11.78(s,1H),8.45(d,J=8.8Hz,2H),8.02(d,J=8.8Hz,2H),7.75(t,J=8.0Hz,1H),7.6 2(dd,J=7.6,1.1Hz,1H),7.55(d,J=1.6Hz,1H),7.34–7.30(m,2H),3.79–3.69(m,2H),3.50–3.39(m,2H),3.19–3.04(m,4H). 13 C NMR (126MHz, DMSO-d6) δ191.25,180.74,166.49,161.33,161.10,150.12,143.52,140.95,137.50,133.6 7,133.01,129.10,124.73,124.53,121.86,119.37,117.26,116.32,115.75,46.16,45.25.IR(KBr)ν / cm -1 :1772,1677,1642,1529,1470,1351,1281,1143,937,743; HRMS(ESI)calcd for C 25 H 20 N3O9S[M+H] + 538.0915, found 538.0906.

[0099] Rhein-piperazine-sulfonamide hybrid 5g: yellow solid, yield 71%, mp 166.3–167.1℃; 1 H NMR (400MHz, DMSO-d6) δ11.86(s,2H),7.77(t,J=8.0Hz,1H),7.65(d,J=7.5Hz,1H),7.61(s,1H),7.51(d,J=4.0Hz,1H ),7.48(d,J=4.1Hz,1H),7.39(s,1H),7.34(d,J=8.4Hz,1H),3.86–3.72(m,2H),3.55–3.41(m,2H),3.21–3.01(m,4H). 13C NMR(101MHz,DMSO-d6)δ191.32,180.82,166.53,161.35,161.14,143.62,137.52,135.96,133.95,133.6 9,133.08,132.15,124.56,121.99,119.83,119.41,117.39,116.35,115.82,46.03,45.47;IR(KBr)ν / cm -1 :1628,1454,1357,1267,1245,1208,1161,1143,723,588; HRMS(ESI)calcd forC 23 H 18 BrN2O7S2[M+H] + 576.9733, found 576.9736.

[0100] Rhein-piperazine-sulfonamide hybrid 5h: yellow solid, yield 65%, mp 147.6–148.5℃; 1 H NMR (400MHz, CDCl3) δ8.18(d,J=7.9Hz,1H),8.09(d,J=1.7Hz,1H),7.77(t,J=8.0 Hz,1H),7.75(s,1H),7.73(d,J=1.7Hz,1H),7.65(t,J=7.4Hz,1H),7.56(t,J=7.5 Hz,2H),7.42(dd,J=8.0,1.3Hz,1H),7.37(d,J=1.7Hz,1H),3.97–3.74(m,2H),3. 64–3.43(m,2H),3.16–3.04(m,2H),3.02–2.97(m,2H),2.43(s,3H),2.42(s,3H). 13 C NMR (101MHz, CDCl3) δ181.29,180.19,169.37,169.17,167.00,150.51,150.24,140.80,135.36,135.07,134.72,13 4.23,133.46,130.72,129.49,128.95,127.74,126.49,125.65,125.58,123.58,45.79,36.05,21.11; IR(KBr)ν / cm -1 :1773,1676,1644,1326,1281,1257,1196,1170,740,577; HRMS(ESI)calcd for C 29 H25 N₂O₉S[M+H] + 577.1275, found 577.1263.

[0101] Rhein-piperazine-sulfonamide hybrid 5i: yellow solid, yield 67%, mp 101.2–102.5 °C; 1 H NMR (400MHz, CDCl3) δ8.20(dd,J=7.8,1.3Hz,1H),8.10(d,J=1.7Hz,1H),7.78(t,J=7.9Hz,1H),7.62(d,J=8.3Hz,2H),7.43(dd,J=8.0,1.3Hz,1H) ,7.38(d,J=1.7Hz,1H),7.37(s,1H),7.35(s,1H),3.97–3.78(m,2H),3.6 2–3.45(m,2H),3.12–2.94(m,4H),2.46(s,3H),2.43(s,3H),2.43(s,3H). 13 CNMR(101MHz, CDCl3)δ181.36,180.22,169.41,169.22,167.02,150.55,150.28,144.45,140.87,135.11,134.76,134.26, 132.25,130.77,130.16,129.00,127.84,126.52,125.70,125.62,123.61,47.04,46.07,21.73,21.71,21.15; IR(KBr)ν / cm -1 :1774,1677,1643,1326,1281,1257,1196,1165,725,548; HRMS(ESI)calcd forC 30 H 26 N₂NaO₉S[M+Na] + 613.1251, found 613.1238.

[0102] Rhein-piperazine-sulfonamide hybrid 5j: yellow solid, yield 69%, mp 173.6–174.1℃; 1H NMR (400MHz, CDCl3) δ8.17(dd,J=8.3,0.8Hz,1H),8.09(d,J=1.7Hz,1H),7.76(t,J=7.9Hz,1H),7.66(d,J=8.9Hz,2H),7.41(dd,J=8.1, 1.3Hz,1H),7.37(d,J=1.7Hz,1H),7.01(d,J=8.9Hz,2H),3.87(s,3H),3.58–3.46(m,2H),3.18–2.81(m,6H),2.42(s,3H),2.41(s,3H). 13 C NMR (101MHz, CDCl3) δ181.25,180.15,169.31,169.12,166.91,163.44,150.44,150.18,140.83,135.02,134.67,134.1 8,130.66,129.88,128.90,126.65,126.41,125.57,125.52,123.54,114.60,55.74,53.54,45.77,21.05; IR(KBr)ν / cm -1 :1773,1677,1643,1595,1326,1281,1258,1196,1161,558; HRMS(ESI)calcd for C 30 H 26 N2NaO 10 S[M+Na] + 629.1200, found 629.1194.

[0103] Rhein-piperazine-sulfonamide hybrid 5K: yellow solid, yield 68%, mp 161.1–162.3 °C; 1 H NMR (400MHz, DMSO-d6) δ8.12(dd,J=7.8,1.3Hz,1H),8.06(d,J=1.7Hz,1H),7.94(t,J=7.9Hz,1H),7.75(tt,J=9.1,2.3Hz,1H),7.66(d ,J=1.6Hz,1H),7.63(dd,J=8.1,1.3Hz,1H),7.54–7.49(m,2H),3.81–3.66(m,2H),3.47–3.37(m,2H),3.21–3.01(m,4H),2.39(s,6H). 13C NMR (101MHz, DMSO-d6) δ180.95,180.26,169.06,168.96,166.14,162.52(d,J=252.5Hz), 162.40(d,J=252.5Hz), 149.52(d,J=3.0Hz), 141.57,135.47,134.53,134.09,130.63,128.38,125.69,125.22,124.96,123.21,111.40(d,J=27.3Hz),109.26,46.26,45.38,20.82; 19 F NMR(376MHz,DMSO-d6)δ-105.92; IR(KBr)ν / cm -1 :1678,1644,1606,1441,1366,1282,1257,1196,1168,594; HRMS(ESI)calcd for C 29 H 22 F₂N₂NaO₉S[M+Na] + 635.0906, found 635.0901.

[0104] Rhein-piperazine-sulfonamide hybrid 5L: yellow solid, yield 47%, mp 177.6–176.4℃; 1 H NMR (400MHz, DMSO-d6) δ8.11(d,J=7.7Hz,1H),8.08(s,1H),7.93(t,J=7.9Hz,1H),7.73–7.67(m,3H),7.63(d,J=8. 0Hz,1H),7.32(d,J=8.8Hz,1H),3.91(s,3H),3.76–3.65(m,2H),3.40–3.30(m,4H),3.24–3.15(m,2H),2.39(s,6H). 13 C NMR(101MHz,DMSO-d6)δ180.94,180.24,169.05,168.96,166.19,155.68,149.55,149.53,141.66,135.45,134.61,134.51,134.06, 130.61,129.74,128.45,127.07,125.69,125.23,124.94,123.80,123.16,115.39,56.55,46.90,45.36,20.83,20.79; IR(KBr)ν / cm -1:1766,1627,1363,1280,1207,1162,1112,944,588; HRMS(ESI)calcd for C 30 H 25 ClN2NaO 10 S[M+Na] + 663.0811, found663.0811.

[0105] Rhein-piperazine-sulfonamide hybrid 5m: brown solid, 50% yield, mp 133.9–135.1℃; 1 H NMR (400MHz, DMSO-d6) δ8.46(d,J=8.4Hz,2H),8.09(d,J=7.8Hz,1H),8.03(d,J=2.2Hz,2H),8.00(s,1H),7.91(t,J=7 .9Hz,1H),7.63(s,1H),7.61(d,J=7.9Hz,1H),3.84–3.67(m,2H),3.52–3.37(m,2H),3.23–3.00(m,4H),2.39(s,6H). 13 C NMR(101MHz,DMSO-d6)δ180.93,180.23,169.08,168.98,166.15,150.16,149.56,141.50,140.93,135.45,134.51,134 .05,130.63,129.18,128.41,125.69,125.18,124.95,124.81,123.18,54.91,46.29,45.34,20.81,20.79; IR(KBr)ν / cm -1 :1774,1677,1531,1351,1272,1195,1014,937,599; HRMS(ESI)calcd for C 29 H 23 N3NaO 11 S[M+Na] + 644.0945, found 644.0945.

[0106] Rhein-piperazine-sulfonamide hybrid 5n: yellow solid, yield 65%, mp 122.5–123.6℃; 1H NMR (400MHz, CDCl3) δ8.19(d,J=7.8Hz,1H),8.12(d,J=1.7Hz,1H),7.77(t,J=7.9Hz,1H),7.42(d,J=8.4 Hz,2H),7.28(d,J=4.0Hz,1H),7.14(d,J=4.0Hz,1H),3.96–3.47(m,4H),3.22–2.97(m,4H),2.43(s,6H). 13 C NMR (101MHz, CDCl3) δ181.36,180.22,169.41,169.22,167.02,150.55,150.28,144.45,140.87,135.11,134.76,134.2 6,132.25,130.77,130.16,129.00,127.84,126.52,125.70,125.62,123.61,47.04,46.07,21.73,21.15; IR(KBr)ν / cm -1 :1773,1677,1325,1281,1257,1240,1196,1162,1144,589; HRMS(ESI)calcd forC 27 H 21 BrN2NaO9S2[M+Na] + 682.9764, found 682.9766.

[0107] In vitro cytotoxicity of target compound 5:

[0108] HepG2, MCF-7, A549, and LO2 cells in logarithmic growth phase were seeded into 96-well plates at 5000 cells per well, for a total volume of 100 μL. 100 μL of PBS was added to the 36 wells at the edge of the plate. Cells were incubated at 37°C, 5% CO2 for 24 h. After cell attachment, the culture medium was aspirated. For the experimental groups, five drug concentration gradients (0.01 μM, 0.1 μM, 1 μM, 10 μM, and 100 μM) were added in three replicates. The control group received 100 μL of culture medium. Cells were incubated at 37°C, 5% CO2 for 48 h. MTT was then added to the culture medium, and the cells were incubated for another 4 h. The supernatant was aspirated, and 100 μL of LDMSO was added and shaken to dissolve the cells. The absorbance was measured at 570 nm using a microplate reader. The inhibition rate was calculated based on the absorbance, and the IC50 was calculated using the least squares method with GraphPadPrism software. 50 Values. Each experiment was performed in triplicate. The results are shown in Table 1.

[0109] Table 1. Cytotoxicity and selectivity of target compound 5 against HepG2, MCF-7, A549, and LO2.

[0110]

[0111] SI = IC50 of normal human hepatocytes (LO2). 50 IC50 of human lung cancer cells A549 50 .

[0112] Table 1 shows that hybrid 5 exhibits superior cytotoxicity against HepG2 liver cancer cells, MCF-7 breast cancer cells, and A549 lung cancer cells compared to the parent Rhein, with a particularly strong inhibitory effect on A549 lung cancer cells. Structure-activity relationship analysis indicates that the introduction of the sulfonamide scaffold effectively enhances the in vitro cytotoxicity of the parent Rhein. Hybrid 5j (IC50) 50 =7.00 μM) was the most toxic to A549 lung cancer cells, approximately equal to that of the parent Rhein (IC50). 50 =142.23 μM) 20 times more selective than the hybrid 5e (SI≈14, LO2 / A549), which was superior to Rhein (SI≈1) and the positive control drug DOX (SI≈0).

[0113] Using rhein (the parent compound) and the clinical anticancer drug doxorubicin (DOX) as references, the in vitro cytotoxicity of hybrid 5 was detected by the MTT assay. The inhibitory rates of different concentrations of hybrid 5 on human lung cancer cells A549 and normal human liver cells LO2 were as follows: Figure 45 As shown, the inhibition rate gradually increases with increasing drug concentration.

[0114] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A rhein-piperazine-sulfonamide hybrid, characterized in that, The rhein-piperazine-sulfonamide hybrid is a compound of formula (I) or a pharmaceutically acceptable salt thereof: （I） Among them, R 1 Selected from hydrogen or acetyl groups; R 2 Selected from one of the following groups: 。 2. The method for preparing the rhein-piperazine-sulfonamide hybrid according to claim 1, characterized in that, The synthesis route is as follows: The preparation method includes: Rhein 1 or diacerein 2 reacts with oxalyl chloride in the presence of a catalyst and a polar aprotic solvent to give intermediate 3. Intermediate 3 and piperazine-sulfonamide derivative 4 were subjected to N-acylation in the presence of an acid-binding agent and a polar aprotic solvent to give compound 5, namely the compound shown in formula (I); Among them, R in piperazine-sulfonamide derivative 4 2 The definition is as described in claim 1.

3. The method for preparing the rhein-piperazine-sulfonamide hybrid as described in claim 2, characterized in that, The catalyst is N,N-dimethylformamide; the polar aprotic solvent is dichloromethane; and the acid-binding agent is triethylamine.

4. A pharmaceutical composition, characterized in that, Includes the rhein-piperazine-sulfonamide hybrid as described in claim 1.

5. A pharmaceutical preparation, characterized in that, It includes the rhein-piperazine-sulfonamide hybrid as described in claim 1, and at least one pharmaceutically acceptable excipient or carrier.

6. The pharmaceutical formulation as described in claim 5, characterized in that, The preparation is a tablet, capsule, granule or injection.

7. The use of the rhein-piperazine-sulfonamide hybrid of claim 1, the pharmaceutical composition of claim 4, or the pharmaceutical preparation of claim 5 or 6 in the preparation of an anticancer drug, wherein the cancer is human lung cancer.

8. The use of the rhein-piperazine-sulfonamide hybrid of claim 1, the pharmaceutical composition of claim 4, or the pharmaceutical formulation of claim 5 or 6 in the preparation of an anticancer drug, wherein, The R 2 Selected from one of the following groups: The cancer mentioned is either human liver cancer or human breast cancer.

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