Ibuprofen-diclometisone conjugate, its preparation method and application

Abstract

The application discloses a kind of ibuprofen-desloratadine conjugates and its preparation method and application, with structure shown in formula I.The application further provides the preparation method of the conjugate, including the reaction of ibuprofen and desloratadine in the presence of condensation reagent, and the target product is obtained after purification.Cell activity test shows that the half inhibitory concentration (IC 50 ) of the conjugate on human hepatoma HepG2 cells and Hep3B cells is significantly better than desloratadine and positive control drug 5-fluorouracil.The conjugate of the application integrates the double-target anti-tumor mechanism of two drugs, and can be used for preparing anti-hepatoma drugs.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to an ibuprofen-desloratadine conjugate, its preparation method, and its application. Background Technology

[0002] Liver cancer is a highly prevalent malignant tumor worldwide, with hepatocellular carcinoma accounting for 75%–85% of primary liver cancers. It ranks as the sixth most common malignant tumor and the third leading cause of cancer-related death globally. my country has the heaviest burden of liver cancer, accounting for over 40% of both new cases and deaths globally. Liver cancer has an insidious onset, with approximately 70% of patients diagnosed at an advanced stage, and a 5-year survival rate of only 12.1%. Traditional radiotherapy and chemotherapy cause significant damage to normal cells, and patients in the middle and late stages often experience severe adverse reactions. Therefore, developing novel, highly effective, and low-toxicity anti-liver cancer drugs is of significant clinical importance.

[0003] In recent years, drug retargeting (i.e., exploring new indications for already marketed drugs) has become an important direction in anti-tumor drug development, offering advantages such as shorter development cycles, lower costs, and established safety profiles. Desloratadine, a third-generation antihistamine, has recently been found to inhibit the myristylation modification of the VILIP3 protein by targeting N-myristyltransferase 1, thereby blocking the NF-κB signaling pathway and inhibiting the proliferation, migration, and invasion of liver cancer cells. A patent (CN110755431B) discloses the application of desloratadine in the preparation of anti-liver cancer drugs. However, when used alone, desloratadine's inhibitory activity against liver cancer cells still needs improvement, requiring higher concentrations to achieve the desired effect.

[0004] Ibuprofen, a classic nonsteroidal anti-inflammatory drug, has recently been found to have anti-tumor effects. It can reduce inflammatory factors in the tumor microenvironment by inhibiting cyclooxygenase-2 activity and simultaneously regulate tumor cell apoptosis pathways. Studies have shown that ibuprofen derivatives also exhibit certain inhibitory activity against liver cancer cells. However, when ibuprofen is used alone, its targeting and inhibitory effects on liver cancer cells are limited. Simply combining desloratadine with ibuprofen presents problems such as pharmacokinetic mismatch and insufficient synergistic effect, making it difficult to fully realize the synergistic anti-tumor effect of both drugs.

[0005] Currently, there are no reports in existing technologies of covalently linking desloratadine and ibuprofen via chemical bonds to form a single molecule compound, nor are there any studies on the use of such conjugates for the treatment of liver cancer. Therefore, developing a novel conjugate that can simultaneously integrate the anti-tumor mechanisms of two drugs, improve pharmacokinetic synchronicity, and enhance anti-liver cancer activity has significant innovative value and clinical potential. Summary of the Invention

[0006] This invention provides an ibuprofen-desloratadine conjugate, its preparation method, and its applications. The conjugate covalently links ibuprofen and desloratadine via an amide bond, forming a novel chemical entity. In vitro antitumor experiments show that this conjugate exhibits significantly superior inhibitory activity against HepG2 and Hep3B hepatocellular carcinoma cells compared to desloratadine and the positive control drug 5-fluorouracil, while exhibiting extremely low toxicity to normal hepatocytes. It can be used to prepare highly effective and low-toxicity anti-hepatocellular carcinoma drugs.

[0007] On the one hand, the present invention provides an ibuprofen-deloratadine coupling compound, which adopts the following technical solution: An ibuprofen-deloratadine coupling compound having the structure shown in Formula I: Formula I; Preferably, ibuprofen and desloratadine are coupled via amide bonds.

[0008] Preferably, the coupling agent comprises a pharmaceutically acceptable salt selected from: hydrochloride, hydrobromide, sulfate, hydrogen sulfate, nitrate, phosphate, hydrogen phosphate, formate, acetate, propionate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, succinate, gluconate, methanesulfonate, ethylsulfonate, benzenesulfonate, and p-toluenesulfonate.

[0009] On the other hand, the present invention also provides a method for preparing an ibuprofen-deloratadine conjugate, which adopts the following technical solution: A method for preparing an ibuprofen-deloratadine conjugate includes the following steps: 1) Dissolve ibuprofen and desloratadine in an organic solvent, and add a condensing agent under nitrogen protection to carry out the reaction; 2) After the reaction is complete, the ibuprofen-deloratadine conjugate is obtained by filtration, washing and drying, or by washing, drying and column chromatography purification.

[0010] Preferably, the organic solvent is acetone or dichloromethane.

[0011] Preferably, the condensation reagent includes EDCI.HCl.

[0012] The present invention also provides a pharmaceutical composition comprising the above-described ibuprofen-deloratadine conjugate or a pharmaceutically acceptable salt thereof.

[0013] The present invention also provides a pharmaceutical formulation comprising the above-described ibuprofen-desloratadine conjugate and at least one pharmaceutically acceptable excipient or drug carrier. Preferably, the pharmaceutical preparation is selected from tablets, capsules, pills or injections.

[0014] The present invention also provides the use of ibuprofen-deloratadine conjugate or the above-described pharmaceutical composition or the above-described pharmaceutical preparation in the preparation of an anti-liver cancer drug.

[0015] In summary, the beneficial effects of the present invention are as follows: This invention provides an ibuprofen-desloratadine conjugate, its preparation method, and its application. For the first time, ibuprofen and desloratadine are covalently linked via an amide bond to prepare a novel chemical entity—the ibuprofen-desloratadine conjugate. Cellular experiments show that this conjugate exhibits a half-maximal inhibitory concentration (IC50) against HepG2 liver cancer cells. 50 The concentration of the active ingredient was 5.69 μmol / L, significantly better than desloratadine (25.13 μmol / L) and the positive control drug 5-fluorouracil (226.31 μmol / L), and the half-maximal inhibitory concentration (IC50) against Hep3B liver cancer cells was significantly higher. 50 The concentration of the active ingredient was 9.06 μmol / L, which was superior to desloratadine (23.89 μmol / L) and the positive control drug 5-fluorouracil (43.25 μmol / L), indicating a significant improvement in anti-hepatocellular carcinoma activity. Simultaneously, the IC50 of this conjugate against LO2 cells in normal hepatocytes was [not specified in the original text]. 50 With a concentration greater than 200 μmol / L, it exhibits excellent tissue selectivity and low toxicity, overcoming the shortcomings of traditional chemotherapy drugs that cause significant damage to normal cells.

[0016] The conjugate of this invention utilizes a degradable amide bond, allowing for the slow release of the original drug in vivo. It combines the advantages of a prodrug, improving the pharmacokinetic synchronicity of desloratadine and ibuprofen, increasing drug concentration at the tumor site, and reducing the side effects of monotherapy. This conjugate integrates the dual-target antitumor mechanism of desloratadine targeting the NMT1 / NF-κB pathway and ibuprofen inhibiting the COX-2 / inflammatory pathway, achieving synergistic effects and providing a promising candidate compound for the development of novel anti-liver cancer drugs. Attached Figure Description

[0017] Figure 1 The proton NMR spectrum of ibuprofen-deloratadine conjugate ( 1 H-NMR spectrum; Figure 2 IC50 of ibuprofen-deloratadine conjugate against human hepatocellular carcinoma cells HepG2 50 Fitted plot; Figure 3 IC50 of ibuprofen-deloratadine conjugate against human hepatocellular carcinoma cells Hep3B 50 Fitted plot; Figure 4 Desloratadine's IC50 response to human HepG2 liver cancer cells 50 Fitted plot; Figure 5 IC50 of desloratadine against human hepatocellular carcinoma cells Hep3B 50 Fitted plot; Figure 6 IC50 of 5-fluorouracil against human hepatocellular carcinoma cells HepG2 50 Fitted plot; Figure 7 IC50 of 5-fluorouracil against human hepatocellular carcinoma cells Hep3B 50 Fitted plot. Detailed Implementation

[0018] The present invention will be further described in detail below with reference to the embodiments.

[0019] Example Example 1 The specific steps for preparing an ibuprofen-deloratadine conjugate are as follows: Dissolve 2.08 g ibuprofen (10.0 mmol) and 3.11 g desloratadine (10.0 mmol) in 100 mL of acetone and place the solution in a 250 mL round-bottom reaction flask. Under nitrogen protection at room temperature, add 2.88 g EDCI·HCl (15.0 mmol). After the addition is complete, stir the mixture at this temperature for 30 min. Filter the solution, wash with 25 mL of acetone, and dry under vacuum at 60 °C for 2.0 h to obtain the ibuprofen-desloratadine conjugate (4.46 g, 89%).

[0020] Example 2 The specific steps for preparing an ibuprofen-deloratadine conjugate are as follows: Dissolve 2.29 g ibuprofen (11.0 mmol) and 3.11 g desloratadine (10.0 mmol) in 100 mL of acetone and place the solution in a 250 mL round-bottom reaction flask. Under nitrogen protection at room temperature, add 3.84 g EDCI·HCl (20.0 mmol). After the addition is complete, stir the mixture at this temperature for 30 min. Filter the solution, wash with 25 mL of acetone, and dry under vacuum at 60 °C for 2.0 h to obtain the ibuprofen-desloratadine conjugate (4.61 g, 92%).

[0021] Example 3 The specific steps for preparing an ibuprofen-deloratadine conjugate are as follows: Dissolve 2.08 g ibuprofen (10 mmol) and 3.11 g desloratadine (10.0 mmol) in 80 mL of dichloromethane. Add 2.88 g EDCI·HCl (15.0 mmol) under nitrogen protection at room temperature. After the addition is complete, stir and maintain the temperature for 2.0 h. After the reaction is complete, the reaction solution is washed successively with 100 mL saturated sodium bicarbonate solution and 100 mL saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain ibuprofen-desloratadine conjugate (4.11 g, 82%).

[0022] Example 4 The specific steps for preparing an ibuprofen-deloratadine conjugate are as follows: Dissolve 2.50 g ibuprofen (10 mmol) and 3.11 g desloratadine (10.0 mmol) in 80 mL of dichloromethane. Add 3.46 g EDCI·HCl (18.0 mmol) under nitrogen protection at room temperature. After the addition is complete, stir and maintain the temperature for 2.0 h. After the reaction is complete, the reaction solution is washed successively with 100 mL saturated sodium bicarbonate solution and 100 mL saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain ibuprofen-desloratadine conjugate (4.11 g, 82%).

[0023] Test case Test Example 1 The structures of the ibuprofen-deloratadine conjugates prepared in Examples 1-4 were confirmed by nuclear magnetic resonance spectroscopy (NMR).

[0024] The final 1H NMR data of the ibuprofen-deloratadine conjugate are as follows: Figure 1 As shown: 1 H NMR (600 MHz, DMSO) δ 8.30 (ddd, J = 44.1, 27.1, 3.3 Hz, 1H), 7.55 (dd, J = 10.0, 7.6Hz, 1H), 7.28 (d, J = 11.5 Hz, 1H), 7.22 – 7.10 (m, 5H), 7.10 – 7.01 (m, 2H), 4.09 – 3.95 (m, 1.5H), 3.65 – 3.52 (m, 1.5H), 3.48 – 3.39 (m, 0.5H), 3.31 –3.08 (m, 3H), 3.04 – 2.86 (m, 0.5H), 2.83 – 2.69 (m, 2H), 2.44 (dd, J= 7.1,3.8 Hz, 1H), 2.39 – 2.03 (m, 3H), 1.99 – 1.70 (m, 2H), 1.64 – 1.33 (m, 1H),1.30 – 1.21 (m, 3H), 0.91 – 0.85 (m, 3H), 0.80 – 0.74 (m, 3H).

[0025] Test Example 2 IC50 of desloratadine-ibuprofen conjugate 50 The specific steps for the measurement are as follows: Experimental cells: Two types of human tumor cells were used in the experiment: HepG2 liver cancer cells (human hepatocellular carcinoma cells) and Hep3B liver cancer cells (human hepatocellular carcinoma cells).

[0026] Test compounds: the desloratadine-ibuprofen conjugate of this application (prepared in Example 1), desloratadine (active pharmaceutical ingredient), ibuprofen (active pharmaceutical ingredient), and 5-fluorouracil (5-FU, positive control). Desloratadine was purchased from Shandong Langnuo Pharmaceutical Co., Ltd., ibuprofen (active pharmaceutical ingredient) was purchased from Shandong Xinhua Pharmaceutical Co., Ltd., and 5-fluorouracil (5-FU, positive control) was purchased from Nanjing Chemical Reagent Co., Ltd.

[0027] Test method: 1. IC 50 Measurement 1) Drug preparation: The compound was dissolved in DMSO to a concentration of 10 mM, and then serially diluted 10-fold to 1 mM, 100 μM, 10 μM, 1 μM, and 0.1 μM. The stock solution concentrations and working concentrations are shown in Table 1. Table 1. Stock solution concentration and working concentration

[0028] 2) Cell culture and plating: Both cell types were cultured in DMEM medium containing 10% FBS. When the confluence reached 75-85%, the cells were digested and counted, and then evenly plated into 96-well plates. The initial cell count for both HepG2 and LO2 was 3000 per well.

[0029] 3) Drug administration: Drug administration was performed 24 hours after cell seeding. Each plate had a DMSO solvent control well and each group had 3 replicates.

[0030] 4) MTT assay: 48 hours after drug administration, add MTT, incubate for 4 hours, discard the supernatant, add 100 μl DMSO, shake on a shaker for 10 minutes, and then measure the OD. 570 With OD 720 The numerical value and cell viability calculation formula are as follows: Cell viability = drug-treated groupOD570 -Medication group OD720 / DMSO group OD570 -DMSO OD720 The IC values ​​of each compound were calculated using Graphpad software. 50 .

[0031] 2. Test Results: (1) Perform IC50 assay on the compound 50 The assay detected the IC50 values ​​of two cell lines tested: HepG2 liver cancer cells and LO2 non-tumorigenic liver epithelial cells. 50 The results are shown in Table 2: Table 2. Half-maximal inhibitory concentrations (IC50) of each tested compound against different cell lines. 50 Comparison Table

[0032] The data analysis in the table above shows that the ibuprofen-desloratadine conjugate has tumor-suppressive activity against HepG2 and Hep3B liver cancer cells. Figure 2 , Figure 3 It is significantly superior to desloratadine ( Figure 4 , Figure 5 ) and positive control drug 5-fluorouracil ( Figure 6 , Figure 7 Ibuprofen showed no inhibitory activity against HepG2 and Hep3B liver cancer cells (Table 2). This indicates that the desloratadine-ibuprofen conjugate of this application can significantly enhance anti-liver cancer activity while achieving good tissue selectivity through dual-target synergistic action, demonstrating excellent clinical development potential.

[0033] The above are all preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Therefore, all equivalent changes made in accordance with the structure, shape and principle of the present invention should be covered within the scope of protection of the present invention.

Claims

1. An ibuprofen-deloratadine coupling compound, characterized in that, It has the structure shown in Equation I: Equation I.

2. The ibuprofen-deloratadine conjugate according to claim 1, characterized in that, The conjugate includes a pharmaceutically acceptable salt selected from: hydrochloride, hydrobromide, sulfate, hydrogen sulfate, nitrate, phosphate, hydrogen phosphate, formate, acetate, propionate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, succinate, gluconate, methanesulfonate, ethylsulfonate, benzenesulfonate, and p-toluenesulfonate.

3. A method for preparing the ibuprofen-deloratadine conjugate according to any one of claims 1 to 2, characterized in that, Includes the following steps: 1) Dissolve ibuprofen and desloratadine in an organic solvent, and add a condensing agent under nitrogen protection to carry out the reaction; 2) After the reaction is complete, the ibuprofen-deloratadine conjugate is obtained by filtration, washing and drying, or by washing, drying and column chromatography purification.

4. The method for preparing an ibuprofen-deloratadine coupling compound according to claim 3, characterized in that, The organic solvent is acetone or dichloromethane.

5. The method for preparing an ibuprofen-deloratadine coupling compound according to claim 3, characterized in that, The condensation reagent includes EDCI.HCl.

6. A pharmaceutical composition, characterized in that, It comprises the ibuprofen-deloratadine conjugate or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 2.

7. A pharmaceutical preparation, characterized in that, It comprises the ibuprofen-deloratadine conjugate according to any one of claims 1 to 2, and at least one pharmaceutically acceptable excipient or drug carrier.

8. The pharmaceutical preparation according to claim 7, characterized in that, The pharmaceutical preparation is selected from tablets, capsules, pills, or injections.

9. The use of the ibuprofen-deloratadine conjugate according to any one of claims 1 to 2, the pharmaceutical composition according to claim 6, or the pharmaceutical preparation according to claim 7 in the preparation of an anti-hepatocellular carcinoma drug.

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