A platinum (IV) prodrug with a novel nf-kappa b inhibitor as ligand, its preparation method and application

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and specifically discloses a platinum (IV) prodrug with a new type of NF-kappa B inhibitor as a ligand as well as a preparation method and application thereof. A new type of NF-kappa B inhibitor is introduced into an axial position of a platinum (IV) complex, and a new type of platinum (IV) antitumor drug with strong targeting, high efficacy and low toxicity is designed and synthesized. The platinum (IV) complex disclosed by the application has good anti-lung cancer activity and low toxicity to human normal liver cell L02 cells. In addition, the prodrug shows good inhibition effect on cisplatin-resistant cell A549 / CDDP. The in-vitro research results show that the platinum (IV) prodrug has potential use in targeted treatment of lung cancer.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and relates to design and synthesis of a chemical prodrug, in particular to a platinum(IV) prodrug taking a new NF-kappa B inhibitor as a ligand and a synthesis method and application thereof. BACKGROUND

[0002] Modern medicine has developed rapidly, and people have overcome many diseases that are difficult to treat, but cancer is still one of the main causes of human death. Platinum drugs, such as cisplatin, carboplatin and oxaliplatin, are considered as first-line chemotherapy drugs due to their low price, good efficacy and wide applicability, and are widely used in the clinical treatment of various solid tumors. Although this kind of anticancer drugs has achieved great results in clinical practice, their serious side effects (such as nephrotoxicity, ototoxicity and bone marrow suppression) and drug resistance limit their application in clinical practice. Therefore, it is urgent to develop a new type of platinum anticancer drug with strong targeting, high efficacy and low toxicity.

[0003] In recent years, it is generally believed that platinum(IV) antitumor complexes can act as prodrugs of platinum(II) antitumor drugs, and can be reduced by biological reducing substances (such as glutathione and ascorbic acid) to release the axial ligand and the corresponding platinum(II) drug. On the one hand, the stability of platinum(IV) complexes is better than that of platinum(II) complexes, thereby improving the opportunity of complete entry into tumor cells or tumor tissues, which is conducive to improving the therapeutic efficacy. On the other hand, the axial position of the platinum(IV) complex can introduce the desired functional groups, thereby enhancing the targeting and anticancer activity. On this basis, some technicians focus on the modification of the axial position of the platinum(IV) complex to design and synthesize a new type of platinum(IV) antitumor drug with strong targeting, high efficacy and low toxicity. SUMMARY

[0004] In view of the deficiencies of the prior art, the purpose of the present application is to provide a platinum(IV) prodrug taking a new NF-kappa B inhibitor as a ligand, which can overcome the shortcomings of existing cisplatin antitumor drugs, such as easy drug resistance and large side effects, and further provides a synthesis method of the platinum(IV) prodrug.

[0005] The present application is realized by the following technical solutions:

[0006] A platinum(IV) prodrug taking a new NF-kappa B inhibitor as a ligand, the structural general formula of the platinum(IV) prodrug is shown in formula 8, formula 9, formula 18, formula 19, formula 20 or formula 21:

[0007]

[0008] Further improved schemes of the present application are:

[0009] The preparation method of the platinum (IV) prodrug compound 8 or 9 with the new type of NF-κB inhibitor as a ligand comprises the following steps: (1) synthesizing compound 3 by reacting compound 1 with compound 2 under the condition of an inorganic base;

[0010] (2) synthesizing compound 4 by reacting compound 3 with bromoethanol under the condition of an inorganic base;

[0011] (3) synthesizing compound 5 or 6 by reacting compound 4 with succinic anhydride or glutaric anhydride under the condition of an organic base;

[0012] (4) synthesizing compound 8 or 9 by reacting compound 5 or 6 with a platinum (IV) intermediate under the action of a condensing agent and an organic base; the synthesis route is shown as follows:

[0013]

[0014] Further, the inorganic base in step (1) is 50% potassium hydroxide aqueous solution;

[0015] And / or, the inorganic base in step (2) is K2CO3;

[0016] And / or, the organic base in step (3) is triethylamine;

[0017] And / or, the condensing agent in step (4) is O-benzotriazole-N,N,N,N-tetramethyluronium tetrafluoroborate (TBTU), and the organic base is triethylamine.

[0018] Further, in step (1), the molar ratio of compound 1: compound 2: inorganic base is 1:1:(10-20);

[0019] And / or, in step (2), the molar ratio of compound 3: bromoethanol: inorganic base is 1:(1-2):(1-5);

[0020] And / or, in step (3), the molar ratio of compound 4: succinic anhydride or glutaric anhydride: organic base is 1:(2-5):(2-5);

[0021] And / or, in step (4), the molar ratio of compound 5 or 6: platinum (IV) intermediate: condensing agent: organic base is 1:(0.5-1.5):(1-3):(1-3).

[0022] Further, the reaction temperature of step (1) is -5-5°C, and the time is 8-16h;

[0023] And / or, the reaction temperature of step (2) is 60-100°C, and the time is 8-16h;

[0024] And / or, the reaction temperature of step (3) is 50-70℃, and the reaction time is 8-16h.

[0025] And / or, the reaction temperature of step (4) is 20-50℃, and the reaction time is 8-16h.

[0026] A further improved scheme of the present application is:

[0027] The preparation method of the platinum (IV) prodrug compound 18 or 19 with the new type of NF-κB inhibitor as a ligand comprises the following steps:

[0028] (1) synthesizing compound 10 by reacting compound 3 with N-Boc-bromoethylamine under the catalysis of an inorganic base;

[0029] (2) synthesizing compound 12 by reacting compound 10 with 4N hydrochloric acid / methanol solution;

[0030] (3) synthesizing compound 14 or 15 by reacting compound 12 with succinic anhydride or glutaric anhydride;

[0031] (4) synthesizing compound 18 or 19 by reacting compound 14 or 15 with a platinum (IV) intermediate under the action of a condensing agent and an organic base;

[0032] The synthesis route is as shown below:

[0033]

[0034] Further, the inorganic base in step (1) is K2CO3; in terms of molar amount, compound 3:N-Boc-bromoethylamine:inorganic base=1:(1-2):(1-5); the reaction temperature is 40-60℃, and the reaction time is 8-16h;

[0035] And / or, in step (2), in terms of molar amount, compound 10:4N hydrochloric acid / methanol solution=1:(0.5-1.5); the reaction temperature is 20-30℃, and the reaction time is 8-16h;

[0036] And / or, in step (3), the organic base is preferably triethylamine; in terms of molar amount, compound 12:succinic anhydride or glutaric anhydride:organic base=1:(2-5):(2-5); the reaction temperature is 50-70℃, and the reaction time is 8-16h;

[0037] And / or, in step (4), in terms of molar amount, compound 14 or 15:platinum (IV) intermediate:condensing agent:organic base=1:(0.5-1.5):(1-3):(1-3); the reaction temperature is 20-50℃, and the reaction time is 8-16h.

[0038] A further improved scheme of the present application is:

[0039] The preparation method of the platinum (IV) prodrug compound 20 or 21 with the new type of NF-κB inhibitor as a ligand comprises the following steps:

[0040] (1) synthesizing compound 11 by reacting compound 3 with N-Boc-bromopropylamine under the catalysis of an inorganic base;

[0041] (2) synthesizing compound 13 by reacting compound 11 with a 4N hydrochloric acid / methanol solution;

[0042] (3) synthesizing compound 16 or 17 by reacting compound 13 with succinic anhydride or glutaric anhydride;

[0043] (4) synthesizing compound 20 or 21 by reacting compound 16 or 17 with a platinum (IV) intermediate under the action of a condensing agent and an organic base;

[0044] The synthesis route is shown as follows:

[0045]

[0046] Further, the inorganic base in step (1) is K2CO3; in terms of molar amount, compound 3:N-Boc-bromopropylamine:inorganic base = 1:(1-2):(1-5); the reaction temperature is 40-60°C, and the time is 8-16h;

[0047] And / or, in step (2), in terms of molar amount, compound 11:4N hydrochloric acid / methanol solution = 1:(0.5-1.5); the reaction temperature is 20-30°C, and the time is 8-16h;

[0048] And / or, in step (3), the organic base is preferably triethylamine; in terms of molar amount, compound 13: succinic anhydride or glutaric anhydride:organic base = 1:(2-5):(2-5); the reaction temperature is 50-70°C, and the time is 8-16h;

[0049] And / or, in step (4), in terms of molar amount, compound 16 or 17: platinum (IV) intermediate:condensing agent:organic base = 1:(0.5-1.5):(1-3):(1-3); the reaction temperature is 20-50°C, and the time is 8-16h.

[0050] Further improvement of the present application is:

[0051] The platinum (IV) prodrug with the new type of NF-κB inhibitor as a ligand in the application of anti-lung cancer

[0052] Compared with the prior art, the present application has the following beneficial effects:

[0053] The present application introduces a NF-κB inhibitor into the axial position of a platinum (IV) intermediate, to obtain a new type of NF-κB inhibitor as a platinum (IV) prodrug of ligand. Studies have shown that the platinum (IV) complex has good anti-lung cancer activity, and the toxicity to human normal liver cells LO2 cells is low. In addition, the prodrug shows good inhibition effect on cisplatin-resistant cell A549 / CDDP. The in vitro study results show that the platinum (IV) prodrug has potential use for targeted treatment of lung cancer. BRIEF DESCRIPTION OF DRAWINGS

[0054] Figure 1 A549 xenograft tumor model for the anti-tumor effect of compound 9 is shown in the figure;

[0055] Wherein, (A) tumor images of mice after treatment with physiological saline (0.9%), CDDP (5.0 mg / kg), 4 (5 mg / kg), CDDP+4 (5+5 mg / kg) and compound 9 (13.9 mg / kg). (B) Tumor volume change after treatment. (C) Change of body weight of mice after treatment. (D) Tumor weight of mice in each group. *p<0.05, **p<0.01.

[0056] Figure 2 A549 / CDDP xenograft tumor model for the tumor effect of compound 9 is shown in the figure;

[0057] Wherein, (A) tumor images of mice after treatment with physiological saline (0.9%), CDDP (5.0 mg / kg), 4 (5 mg / kg), CDDP+4 (5+5 mg / kg) and compound 9 (13.9 mg / kg). (B) Tumor volume. (C) Change of body weight of mice after 21 days of treatment. (D) Tumor weight of mice in each group. *p<0.05, **p<0.01. DETAILED DESCRIPTION

[0058] The present application will be described in detail below in combination with specific examples.

[0059] Abbreviations in the experimental part: DCM, dichloromethane; MeOH, methanol; EA, ethyl acetate; PE, petroleum ether; DMSO, dimethyl sulfoxide; CDCl3, deuterated chloroform; CDDP, cisplatin; DMF, N,N-dimethylformamide; TLC, thin layer chromatography; EtOH, ethanol; Et3N, triethylamine; TBTU, O-benzotriazole-N,N,N,N-tetramethyluronium tetrafluoroborate; HepG-2, human hepatoma cells; HCT-116, human colon cancer cells; MCF-7, human breast cancer cells; A549, human lung cancer cells; L02, human normal liver cells

[0060] Experimental materials and equipment: Unless otherwise specified, all chemicals and solvents were purchased from commercial companies without additional treatment. The 400, 500 or 600 MHz nuclear magnetic resonance spectrometer of Bruker was used to measure 1 H and 13 C nuclear magnetic resonance spectra, and analyzed by MestReNova software. High resolution mass spectrometry (HR-MS) was measured by Agilent LC / MS (6545 Q-TOF) with methanol as solvent. The purity of all target compounds used in the experiment was greater than 95%, determined by HPLC, and the chromatographic column was TC-C18 column (4.6 mm x 250 mm, 5 μm). The cells used in the experiment, such as hepatoma cells (HepG-2), colon cancer cells (HCT-116), breast cancer cells (MCF-7), lung cancer cells (A549), were from American Type Culture Collection, and cisplatin-resistant cells (A549 / CDDP) were purchased from Beijing Union Medical College.

[0061] Example 1: Preparation of compound 3

[0062] Compound 1 (4.2 g, 20 mmol) and compound 2 (3.04 g, 20.0 mmol) were dissolved in MeOH (30 mL), and 50% KOH (15 mL, aq) was slowly added under ice water cooling. After the addition was completed, the reaction was allowed to stand overnight under ice water bath. After the reaction was completed, the pH value was adjusted to 1-2 with 2N HCl solution, and then extracted with DCM (2 x 150 mL). The organic phase was dried with anhydrous sodium sulfate and concentrated to obtain the crude product, which was then purified by silica gel column chromatography to obtain yellow solid 3 (yield: 4.5 g, 65.4%). 1 H NMR (400 MHz, CDC13) δ 7.75 (d, J = 15.5 Hz, 1H), 7.35 (d, J = 15.5 Hz, 1H), 7.31 (d, J = 2.1 Hz, 1H), 7.27 (s, 2H), 7.15-7.12 (m, 1H), 6.88 (d, J = 8.3 Hz, 1H), 3.95 (s, 6H), 3.94 (s, 3H), 3.93 (s, 3H). HR-MS (m / z) (ESI): calcd for C 19 H 20 O6[M+H] + : 345.1338; found: 345.1335.

[0063] Example 2: Preparation of compound 4

[0064] Compound 3 (700 mg, 2.03 mmol) and bromoethanol (380 mg, 3.05 mmol) were dissolved in DMF (5 mL), K2CO3 (560 mg, 4.06 mmol) was added and stirred at 80 °C overnight. After the reaction was completed, DCM (100 mL) was added, washed with water (3 x 200 mL), and the organic phase was dried over anhydrous sodium sulfate, concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain yellow solid 4 (yield: 650 mg, 82.6%). 1 HNMR (400 MHz, CDC13) δ 7.74 (d, J = 15.6 Hz, 1H), 7.32 (d, J = 15.6 Hz, 1H), 7.29 - 7.26 (m, 1H), 7.25 (s, 2H), 7.22 (d, J = 1.9 Hz, 1H), 6.92 (d, J = 8.4 Hz, 1H), 4.20 - 4.17 (m, 2H), 4.00 - 3.96 (m, 2H), 3.94 (s, 6H), 3.93 (s, 3H), 3.91 (s, 3H). HR-MS (m / z) (ESI): calcd for C 21 H 24 O7[M+Na] + : 411.1420; found: 411.1410.

[0065] Example 3: Preparation of compounds 5 and 6

[0066] Compound 4 (600 mg, 1.55 mmol) was dissolved in DMF (5 mL), succinic anhydride (465 mg, 4.65 mmol) or glutaric anhydride (530 mg, 4.65 mmol) was added, and stirred at 60 °C overnight. After the reaction was completed, DCM (100 mL) was added, washed with water (3 x 200 mL), and the organic phase was dried over anhydrous sodium sulfate, concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain product 5 or 6.

[0067] Compound 5, yellow solid (690 mg), yield 91.3%. 1H NMR (400 MHz, DMSO-d6) δ 12.24 (s, 1H), 7.80 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.56 (d, J = 1.8 Hz, 1H), 7.50 - 7.47 (m, 1H), 7.40 (s, 2H), 7.06 (d, J = 8.5 Hz, 1H), 4.38 - 4.36 (m, 2H), 4.28 - 4.26 (m, 2H), 3.90 (s, 6H), 3.84 (s, 3H), 3.76 (s, 3H), 2.57 - 2.54 (m, 2H), 2.49 - 2.47 (m, 2H). HR-MS (m / z) (ESI): calcd for C 25 H 28 O 10 [M+H] + : 489.1761; found: 489.1755.

[0068] Compound 6, yellow solid (680 mg), yield 87.4%. 1 H NMR (400 MHz, DMSO-d6) δ 12.10 (s, 1H), 7.80 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.55 (d, J = 1.8 Hz, 1H), 7.49 - 7.47 (m, 1H), 7.40 (s, 2H), 7.05 (d, J = 8.5 Hz, 1H), 4.39 - 4.37 (m, 2H), 4.29 - 4.27 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.77 (s, 3H), 2.39 - 2.36 (m, 2H), 2.28 - 2.24 (m, 2H), 1.78 - 1.73 (dd, J = 14.8, 7.4 Hz, 2H). HR-MS (m / z) (ESI): calcd for C 26 H 20 O 10 [M+H] + : 503.1917; found: 503.1905.

[0069] Example 4: Preparation of compounds 8 and 9

[0070] Compound 5 (130 mg, 0.266 mmol) or compound 6 (134 mg, 0.266 mmol), TBTU (102 mg, 0.319 mmol), Et3N (32 mg, 0.319 mmol) were dissolved in dry DMF (2 mL), then complex 7 (94 mg, 0.266 mmol) was added, and stirred at 30 °C overnight. After the reaction was completed, DCM (100 mL) was added, washed with water (2 x 150 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was further purified by silica gel column chromatography with MeOH / DCM system to obtain the product 8 or 9.

[0071] Compound 8, yellow solid (95 mg), yield 43.6%. 1 HNMR (500 MHz, DMSO-d6) δ 7.79 (d, J = 15.3 Hz, 1H), 7.69 (d, J = 15.4 Hz, 1H), 7.55 (s, 1H), 7.49 (d, J = 7.8 Hz, 1H), 7.40 (s, 2H), 7.06 (d, J = 8.3 Hz, 1H), 6.31 - 5.98 (s, 6H), 4.39 - 4.35 (m, 2H), 4.31 - 4.28 (m, 2H), 3.90 (s, 6H), 3.84 (s, 3H), 3.76 (s, 3H), 2.55 - 2.50 (m, 4H). 13 C NMR (125 MHz, DMSO-d6) δ 188.44, 179.40, 172.87, 153.36, 151.96, 148.25, 144.70, 142.35, 133.79, 128.04, 124.50, 120.30, 113.82, 112.54, 106.71, 67.29, 63.03, 60.68, 56.77, 56.24, 31.48, 30.33. HR-MS (m / z) (ESI): calcd for C 25 H 33 Cl3N2O 10 Pt[M+H] + : 822.0927; found: 822.0911. Elemental analysis calcd (%) C 25 H 33 Cl3N2O 10 Pt for: C, 36.49; H, 4.04; N, 3.40; found: C, 36.74; H, 4.26; N, 3.02. Purity 98.16% (by HPLC)

[0072] Compound 9, yellow solid (93 mg), yield 41.9%.1 HNMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 15.4 Hz, 1H), 7.69 (d, J = 15.4 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.40 (s, 2H), 7.05 (d, J = 8.3 Hz, 1H), 6.36 - 5.94 (m, 6H), 4.44 - 4.33 (m, 2H), 4.28 - 4.22 (m, 2H), 3.90 (s, 6H), 3.84 (s, 3H), 3.76 (s, 3H), 2.41 (t, J = 7.1 Hz, 2H), 2.29 (t, J = 6.8 Hz, 2H), 1.80 - 1.171 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 188.42, 180.22, 173.41, 153.36, 151.94, 148.25, 144.72, 142.32, 133.79, 128.02, 124.54, 120.26, 113.69, 112.50, 106.68, 67.27, 65.39, 62.91, 60.68, 56.74, 56.24, 33.15, 32.00, 21.25. HR-MS (m / z) (ESI): calcd for C 26 H 35 Cl3N2O 10 Pt[M+H] + : 836.1083; found: 836.1073. Elemental analysis calcd (%) C 26 H 35 Cl3N2O 10 Pt for: C, 37.31; H, 4.22; N, 3.35; found: C, 37.62; H, 4.51; N, 2.98. Purity 98.02% (by HPLC)

[0073] Example 5: Preparation of compounds 10 and 11

[0074] Compound 3 (1.5 g, 4.36 mmol) and N-Boc-bromoethylamine (1.07 g, 4.80 mmol) or N-Boc-3-aminopropyl bromide (1.14 g, 4.80 mmol) were dissolved in anhydrous DMF (10 mL), K2CO3(1.2 g, 8.72 mmol) was added and stirred at 50 °C overnight. After the reaction was completed, DCM (200 mL) was added, washed with water (2 x 200 mL), the organic phase was dried over anhydrous sodium sulfate, concentrated to give the crude product. The crude product was further purified by silica gel column chromatography with MeOH / DCM system to give product 10 or 11.

[0075] Compound 10. Yellow solid (1.8 g), yield 84.9%. 1 H NMR (400 MHz, CDC13) δ 7.72 (d, J = 15.6 Hz, 1H), 7.32 (d, J = 15.5 Hz, 1H), 7.24 - 7.20 (m, 4H), 6.88 (d, J = 8.4 Hz, 1H), 5.21 (s, 1H), 4.12 - 4.09 (m, 2H), 3.92 (s, 6H), 3.91 (s, 3H), 3.89 (s, 3H), 3.61 - 3.50 (m, 2H), 1.42 (s, 9H). HR-MS (m / z) (ESI): calcd for C 26 H 33 NO8 [M+Na] + : 510.2104; found: 510.2102.

[0076] Compound 11. Yellow solid (2.0 g), yield 91.7%. 1 H NMR (400 MHz, CDC13) δ 7.73 (d, J = 15.6 Hz, 1H), 7.30 (d, J = 15.6 Hz, 1H), 7.25 - 7.24 (m, 3H), 7.14 (d, J = 1.9 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 5.49 (s, 1H), 4.16 - 4.13 (m, 2H), 3.94 (s, 6H), 3.92 (s, 6H), 3.40 - 3.36 (m, 2H), 2.06 - 2.02 (m, 2H), 1.44 (s, 9H). HR-MS (m / z) (ESI): calcd for C 27 H 35 NO8 [M+Na] + : 524.2260; found: 524.2259.

[0077] Example 6: Preparation of compounds 12 and 13

[0078] Compound 10 (1.2 g, 2.46 mmol) or 11 (1.23 g, 2.46 mmol) was dissolved in CH3OH (15 mL), ice water cooling, then slowly added 4N HCl / CH3OH solution (5 mL), stirred at room temperature overnight. After the reaction was completed, the solvent was removed under reduced pressure, the crude product was washed with diethyl ether, and further purified by silica gel column chromatography to obtain compounds 12 and 13.

[0079] Compound 12. Yellow solid (450 mg), yield 47.3%. 1 H NMR (600 MHz, DMSO-d6) δ 8.12 (s, 3H), 7.82 (d, J = 15.6 Hz, 1H), 7.71 (d, J = 15.6 Hz, 1H), 7.59 - 7.54 (m, 1H), 7.41 (s, 2H), 7.10 (d, J = 8.4 Hz, 1H), 4.27 - 4.25 (m, 1H), 3.90 (s, 6H), 3.85 (s, 3H), 3.76 (s, 3H), 3.27 - 3.24 (m, 2H). HR-MS (m / z) (ESI): calcd for C 21 H 25 NO6[M+H] + : 388.1760; found: 388.1751.

[0080] Compound 13. Yellow solid (390 mg), yield 40.5%. 1 H NMR (600 MHz, DMSO-d6) δ 7.88 (s, 3H), 7.80 (d, J = 15.5 Hz, 1H), 7.70 (d, J = 15.4 Hz, 1H), 7.53 - 7.50 (m, 2H), 7.40 (s, 2H), 7.06 (d, J = 8.3 Hz, 1H), 4.16 (t, J = 5.8 Hz, 2H), 3.90 (s, 6H), 3.84 (s, 3H), 3.76 (s, 3H), 2.98 (t, J = 15.5 Hz, 2H), 2.06 - 2.03 (m, 2H). HR-MS (m / z) (ESI): calcd for C 22 H 27 NO6[M+H] + : 402.1907; found: 402.1911.

[0081] Example 7: Preparation of compounds 14-17

[0082] Compound 14 was synthesized from compound 12 and succinic anhydride, with the same synthesis method as compound 5.

[0083] Compound 14. Yellow solid (620 mg), yield 82.2%. 1 H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 8.16 (t, J = 5.5 Hz, 1H), 7.80 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.56 (d, J = 1.8 Hz, 1H), 7.47 - 7.46 (m, 1H), 7.41 (s, 2H), 7.05 (d, J = 8.5 Hz, 1H), 4.08 - 4.05 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.36 - 3.42 (m, 2H), 2.45 - 2.42 (m, 2H), 2.37 - 2.33 (m, 2H). HR-MS (m / z) (ESI): calcd for C 25 H 29 NO9[M+H] + :488.1921; found: 488.1914.

[0084] Compound 15 was synthesized from compound 12 with glutaric anhydride, with the same synthetic method as compound 6.

[0085] Compound 15. Yellow solid (680 mg), yield 87.6%. 1 H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.11 (t, J = 5.4 Hz, 1H), 7.81 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.57 (d, J = 1.3 Hz, 1H), 7.47 - 7.46 (m, 1H), 7.41 (s, 2H), 7.04 (d, J = 8.4 Hz, 1H), 4.09 - 4.06 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.46 - 3.35 (m, 2H), 2.24 - 2.19 (m, 2H), 2.16 - 2.12 (m, 2H), 1.76 - 1.68 (m, 2H). HR-MS (m / z) (ESI): calcd for C 26 H 31 NO9[M+H] + :502.2077; found: 502.2076.

[0086] Compound 16 was synthesized from compound 13 with succinic anhydride, with the same synthetic method as compound 5.

[0087] Compound 16. Yellow solid (675 mg), yield 87.0%.1 H NMR (400 MHz, DMSO-d6) δ 12.06 (s, 1H), 7.87 (t, J = 5.4 Hz, 1H), 7.78 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.50 - 7.46 (m, 2H), 7.40 (s, 2H), 7.04 (d, J = 8.4 Hz, 1H), 4.09 - 4.06 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.25 - 3.20 (m, 2H), 2.26 - 2.20 (m, 2H), 2.12 - 2.08 (m, 2H), 1.89 - 1.85 (m, 2H), 1.75 - 1.69 (m, 2H). HR-MS (m / z) (ESI): calcd for C 26 H 31 NO9 [M+H] + : 516.2234; found: 516.2221.

[0088] Compound 17 was synthesized from compound 13 and glutaric anhydride, with the same synthetic method as compound 6.

[0089] Compound 17. Yellow solid (720 mg), yield 90.2%. 1 H NMR (400 MHz, DMSO-d6) δ 12.06 (s, 1H), 7.87 (t, J = 5.4 Hz, 1H), 7.78 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.5 Hz, 1H), 7.50 - 7.46 (m, 2H), 7.40 (s, 2H), 7.04 (d, J = 8.4 Hz, 1H), 4.09 - 4.06 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.25 - 3.20 (m, 2H), 2.26 - 2.20 (m, 2H), 2.12 - 2.08 (m, 2H), 1.89 - 1.85 (m, 2H), 1.75 - 1.69 (m, 2H). HR-MS (m / z) (ESI): calcd for C 27 H 33 NO9 [M+H] + : 516.2234; found: 516.2221.

[0090] Example 8: Preparation of compounds 18-21

[0091] Compound 18 was synthesized from compound 13 and compound 7, with the same synthetic method as compound 8.

[0092] Compound 18. Yellow solid (132 mg), yield 60.2%. 1 HNMR (400 MHz, DMSO-d6) δ 8.16 (t, J = 5.3 Hz, 1H), 7.80 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.4 Hz, 1H), 7.56 (s, 1H), 7.47 (d, J = 8.3 Hz, 1H), 7.41 (s, 2H), 7.05 (d, J = 8.4 Hz, 1H), 6.48 - 5.90 (m, 6H), 4.08 (t, J = 5.7 Hz, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.45 - 3.40 (m, 2H), 2.49 - 2.45 (m, 2H), 2.47 - 2.43 (m, 2H). 13 CNMR (100 MHz, DMSO-d6) δ 188.43, 180.19, 172.51, 153.36, 151.91, 148.43, 144.79, 142.32, 133.79, 128.04, 124.37, 120.18, 113.54, 112.42, 106.69, 67.65, 65.40, 60.69, 56.76, 56.16, 38.69, 32.46, 32.01. HR-MS (m / z) (ESI): calcd for C 25 H 34 Cl3N3O9Pt[M+H] + : 821.1087; found: 821.1097. Elemental analysis calcd (%) C 25 H 34 Cl3N3O9Pt: C, 36.53; H, 4.17; N, 5.11; found: C, 36.87; H, 4.40; N, 4.79. Purity 98.97% (by HPLC)

[0093] Compound 19 was synthesized from compound 15 and compound 7, the synthesis method was the same as compound 8.

[0094] Compound 19. Yellow solid (85 mg), yield 39.3%. 1H NMR (600 MHz, DMSO-d6) δ 8.05 (t, J = 5.2 Hz, 1H), 7.80 (d, J = 15.5 Hz, 1H), 7.69 (d, J = 15.4 Hz, 1H), 7.57 (s, 1H), 7.46 (d, J = 7.9 Hz, 1H), 7.41 (s, 2H), 7.04 (d, J = 8.4 Hz, 1H), 6.40 - 6.01 (m, 6H), 4.09 - 4.07 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.55 - 3.44 (m, 2H), 2.22 - 2.24 (m, 2H), 2.16 - 2.13 (m, 2H), 1.73 - 1.68 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 188.43, 180.57, 172.95, 153.35, 151.93, 148.44, 144.80, 142.32, 133.79, 128.03, 124.44, 120.15, 113.47, 112.42, 106.69, 67.69, 60.69, 56.75, 56.18, 38.59, 36.14, 35.16, 22.22. HR-MS (m / z) (ESI): calcd for C 26 H 36 Cl3N3O9Pt[M+H] + : 835.1243; found: 835.1254. Elemental analysis calcd (%) C 26 H 36 Cl3N3O9Pt: C, 37.35; H, 4.34; N, 5.03; found: C, 34.66; H, 4.60; N, 4.61. Purity 98.69% (by HPLC)

[0095] Compound 20 was synthesized from compound 16 and compound 7 in the same way as compound 8.

[0096] Compound 20. Yellow solid (97 mg), yield 44.9%. 1H NMR (400 MHz, DMSO-d6) δ 7.93 (t, J = 5.3 Hz, 1H), 7.79 (d, J = 15.5 Hz, 1H), 7.70 (d, J = 15.5 Hz, 1H), 7.50 - 7.47 (m, 2H), 7.40 (s, 2H), 7.04 (d, J = 8.4 Hz, 1H), 6.42 - 5.96 (m, 6H), 4.10 - 4.07 (m, 2H), 3.90 (s, 6H), 3.84 (s, 3H), 3.76 (s, 3H), 3.25 - 3.20 (m, 2H), 2.47 - 2.43 (m, 2H), 2.33 - 2.30 (m, 2H), 1.91 - 1.85 (m, 2H). 13 CNMR (100 MHz, DMSO-d6) δ 188.42, 180.29, 172.01, 153.35, 151.92, 148.64, 144.85, 142.29, 133.80, 128.00, 123.97, 120.15, 113.43, 112.30, 106.66, 66.90, 60.68, 56.74, 56.21, 36.36, 32.55, 32.11, 29.29. HR-MS (m / z) (ESI): calcd for C 26 H 36 Cl3N3O9Pt[M+H] + : 835.1243; found: 835.1265. Elemental analysis calcd (%) C 26 H 36 Cl3N3O9Pt: C, 37.35; H, 4.34; N, 5.03; found: C, 34.69; H, 4.67; N, 4.71. Purity 99.16% (by HPLC)

[0097] Compound 21 was synthesized from compound 17 and compound 7, with the same synthetic method as compound 8.

[0098] Compound 21. Yellow solid (126 mg), yield 55.8%. 1HNMR (400 MHz, DMSO-d6) δ 7.85 - 7.76 (m, 2H), 7.69 (d, J = 15.4 Hz, 1H), 7.50 - 7.46 (m, 2H), 7.40 (s, 2H), 7.04 (d, J = 8.3 Hz, 1H), 6.41 - 5.92 (m, 6H), 4.10 - 4.07 (m, 2H), 3.90 (s, 6H), 3.83 (s, 3H), 3.76 (s, 3H), 3.23 - 3.17 (m, 2H), 2.24 - 2.21 (m, 2H), 2.13 - 2.09 (m, 2H), 1.89 - 1.86 (m, 2H), 1.74 - 1.68 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 188.43, 180.57, 172.45, 153.35, 151.95, 148.63, 144.85, 142.29, 133.80, 128.01, 124.01, 120.16, 113.50, 112.32, 106.67, 66.92, 60.68, 56.74, 56.20, 36.22, 36.16, 35.28, 29.44, 22.27. HR-MS (m / z) (ESI): calcd for C 27 H 38 Cl3N3O9Pt[M+H] + : 849.1400; found: 849.1380. Elemental analysis calcd (%) C 27 H 38 Cl3N3O9Pt: C, 38.15; H, 4.51; N, 4.94; found: C, 38.50; H, 4.83; N, 4.53. Purity 99.01% (by HPLC)

[0099] Example 9: In vitro anti-proliferative activity assay

[0100] 1. Experimental methods

[0101] MTT method was used to evaluate the anti-proliferative activity of the compounds on hepatoma cells (HepG-2), colon cancer cells (HCT-116), breast cancer cells (MCF-7), lung cancer cells (A549), cisplatin-resistant cells (A549 / CDDP) and human normal liver cells (LO2). All cells were inoculated in 96-well plates (3000 cells per well, 2000 μL) and incubated at 37°C in a humidified atmosphere containing 5% CO2 for one night. Then the cells were treated with different concentrations of the tested compounds under the same conditions for 72 hours. After 72 hours of incubation, fresh MTT solution (10 μL) was added and maintained for 4 h. The samples were recorded by an enzyme marker at 570 nm wavelength, and the IC 50 values were calculated by SPSS software.

[0102] Table 1 Toxicity data of compounds on several human tumor cell lines

[0103]

[0104]

[0105] a CDDP, cisplatin; b CDDP combined with 4 (1:1, n / n); c CDDP combined with 12 (1:1, n / n); d CDDP combined with 13 (1:1, n / n). e IC 50 , the values are expressed as the mean ± SD of three independent experiments.

[0106] Table 2 Toxicity data of 9 on cisplatin-resistant cells and normal cells

[0107]

[0108] a CDDP, cisplatin; b IC 50 , the values are expressed as the mean ± SD of three independent experiments. c RF, resistance factor = IC 50 (A549 / CDDP) / IC 50 (A549). d SI, selectivity index = IC 50 (LO2) / IC 50 (A549).

[0109] 2. Experimental results

[0110] As shown in Table 1, compounds 8, 9 and 18-21 have better anti-tumor activity compared with the corresponding positive drugs. Among them, compound 9 (IC50 = 0.29-0.73 μM) into platinum (IV) intermediate 7, IC 50 = 4.18-6.20 μM) and 4 group (IC 50 = 3.67-4.79 μM) compared with 9, the anti-proliferation activity of 9 on A549 cells was 14.41 times and 12.65 times higher than CDDP and 4, respectively.

[0111] As shown in Table 2, the IC 50 values of 4 and 9 on A549 / CDDP cells were 5.14 and 0.51 μM, respectively, which were better than the anti-cancer effect of CDDP, wherein compound 9 showed good anti-tumor activity, and the anti-proliferation activity of compound 9 on A549 / CDDP cells was about 63.06 times of CDDP. In addition, the RF values of 4 and 9 on A549 / CDDP were 5.49 times and 4.36 times lower than CDDP, which indicated that compounds 4 and 9 could effectively overcome CDDP resistance. At the same time, the selectivity index (SI) of compound 9 was 14.86, which was much higher than other groups, indicating that compound 9 had good selectivity on cancer cells. In summary, compound 9 showed good anti-tumor activity on A549 and A549 / CDDP cells in vitro, and had low toxicity on human normal stem cells, so compound 9 was selected as a representative compound for further study.

[0112] Example 10: In vivo anti-tumor activity experiment

[0113] 1. Model establishment

[0114] 5-week-old BALB / c female nude mice, 25 in total, were purchased from Hangzhou Qizhen Experimental Animal Technology Co., Ltd., and were raised under pathogen-free conditions. A549 cells (1 x 10 7 / mouse) were subcutaneously inoculated on the right side of the mice to establish A549 xenograft tumor model. When the tumor volume reached about 100 mm 3 , the 25 mice were randomly divided into 5 groups: normal saline treatment group, CDDP treatment group (5 mg / kg, once a week), 4 treatment group (5 mg / kg, twice a day), CDDP / 4 combination treatment group (5+5 mg / kg, once a week). The tumor volume and body weight changes of the mice were recorded every 2 days after treatment of the test compounds, and the tumor volume was calculated as (length x width 2 ) / 2. After 21 days of treatment, the mice were sacrificed, and the tumors were removed. The tumor inhibition rate was calculated according to the drug treatment results. The A549 / CDDP xenograft tumor model: the method was the same as A549.

[0115] 2. Experimental results

[0116] AsFigure 1 As shown in Table 2, CDDP could effectively inhibit the tumor growth in A549 xenograft tumor model, and the inhibition rate reached 60.4% compared with the control group; compound 4 showed moderate in vivo anti-tumor activity (inhibition rate was 45.7%). Notably, the inhibition effect of the combination group CDDP / 4 on A549 xenograft tumor was stronger than that of CDDP and 4 administration groups, and the tumor inhibition rate reached 68.5%, indicating that drug combination therapy could improve the treatment efficacy. Interestingly, compound 9 (72.7%) could effectively inhibit the growth of A549 xenograft tumor, and the anti-cancer activity was better than that of single administration group or combination administration group. Although CDDP or CDDP / 4 showed better in vivo anti-cancer activity, but the toxic side effects were also obvious, such as Figure 1 As shown in Table 3, the body weight of mice in CDDP group or CDDP / 4 group decreased significantly, while the body weight of mice in compound 9 group was not significantly affected. In order to further study whether compound 9 could overcome cisplatin resistance, we evaluated the inhibition effect of compound 9 on A549 / CDDP xenograft tumor, and cisplatin as a positive drug. As shown in Table 4, Figure 2 As shown in Table 4, the inhibition effect of cisplatin on A549 / CDDP xenograft tumor was significantly lower than that of the sensitive type, and the inhibition rate (29.4%) decreased significantly, and the inhibition effect of compound 4 (36.2%) on A549 / CDDP xenograft tumor was not significantly changed. Compared with the CDDP group, the combination group CDDP / 4 showed stronger anti-tumor activity, and the tumor inhibition rate was 54.9%. Interestingly, compound 9 (63.7%) showed good inhibition effect on A549 / CDDP xenograft tumor, which was stronger than that of single drug administration group or combination administration group. As shown in Table 5, Figure 1 As shown in Table 5, the body weight of mice in CDDP group or CDDP / 4 group changed significantly, showing certain toxic side effects; the body weight of mice in compound 9 group was not significantly changed. Figure 2 As shown in Table 5, the body weight of mice in CDDP group or CDDP / 4 group changed significantly, showing certain toxic side effects; the body weight of mice in compound 9 group was not significantly changed. The in vivo experimental results showed that compound 9 could effectively inhibit the tumor growth in A549 or A549 / CDDP xenograft tumor model, and had no obvious effect on the body weight of mice, indicating that compound 9 had the effects of low toxicity, high efficiency and overcoming cisplatin resistance, and could be used as a potential candidate drug for treating lung cancer.

[0117] The above embodiments only serve to illustrate the technical concept and characteristics of the present application, and the purpose is to enable those skilled in the art to understand the content of the present application and implement it, and cannot limit the protection scope of the present application. Any equivalent transformation or modification made according to the spirit and essence of the present application should be covered within the protection scope of the present application.

Claims

1. A platinum (IV) prodrug with an NF-κB inhibitor as a ligand, characterized in that, The general structural formula of the platinum (IV) prodrug is shown in Formula 8, Formula 9, Formula 18, Formula 19, Formula 20 or Formula 21:

2. A method for preparing a platinum (IV) prodrug with an NF-κB inhibitor as a ligand as described in claim 1, characterized in that, Includes the following steps; (1) React compound 1 and compound 2 under inorganic alkaline conditions to synthesize compound 3; (2) Compound 3 was reacted with bromoethanol under inorganic alkaline conditions to synthesize compound 4; (3) React compound 4 with succinic anhydride or glutaric anhydride under organic base conditions to synthesize compound 5 or 6; (4) React compound 5 or 6 with compound 7 under the action of a condensing agent and an organic base to obtain compound 8 or 9; The synthesis route is shown below:

3. The preparation method according to claim 2, characterized in that: The inorganic base mentioned in step (1) is a 50% potassium hydroxide aqueous solution; And / or, the inorganic base mentioned in step (2) is K2CO3; And / or, the organic base described in step (3) is triethylamine; And / or, the condensing agent in step (4) is O-benzotriazole-N,N,N,N-tetramethylurea tetrafluoroborate (TBTU), and the organic base is triethylamine.

4. The preparation method according to claim 2, characterized in that: In molar quantities, in step (1), compound 1: compound 2: inorganic base = 1:1:(10~20); And / or, in step (2), compound 3: bromoethanol: inorganic base = 1: (1-2): (1-5); And / or, in step (3), compound 4: succinic anhydride or glutaric anhydride: organic base = 1: (2-5): (2-5); And / or, in step (4), compound 5 or 6: compound 7: condensing agent: organic base = 1: (0.5 to 1.5): (1 to 3): (1 to 3).

5. The preparation method according to claim 2, characterized in that: The reaction temperature in step (1) is -5 to 5°C, and the reaction time is 8 to 16 hours. And / or, the reaction temperature in step (2) is 60–100°C and the time is 8–16 h; And / or, the reaction temperature in step (3) is 50–70°C and the reaction time is 8–16 h; And / or, the reaction temperature of step (4) is 20-50℃ and the time is 8-16h.

6. A method for preparing a platinum (IV) prodrug with an NF-κB inhibitor as a ligand as described in claim 1, characterized in that, Includes the following steps; (1) Compound 3 was reacted with N-Boc-bromoethylamine to synthesize compound 10 under inorganic base catalysis; (2) Compound 10 was reacted with 4N hydrochloric acid / methanol solution to synthesize compound 12; (3) React compound 12 with succinic anhydride or glutaric anhydride to synthesize compound 14 or 15; (4) React compound 14 or 15 with compound 7 under the action of a condensing agent and an organic base to obtain compound 18 or 19; The synthesis route is shown below:

7. The preparation method according to claim 6, characterized in that: The inorganic base mentioned in step (1) is K2CO3; in molar amounts, compound 3: N-Boc-bromoethylamine:inorganic base = 1:(1~2):(1~5); the reaction temperature is 40~60℃ and the time is 8~16h; And / or, in step (2), the compound is 10:4N hydrochloric acid / methanol solution = 1:(0.5~1.5) in molar amounts; the reaction temperature is 20~30℃ and the time is 8~16h; And / or, in step (3), in molar amounts, compound 12: succinic anhydride or glutaric anhydride = 1:(2-5); the reaction temperature is 50-70℃ and the time is 8-16h; And / or, in step (4), in molar amounts, compound 14 or 15: compound 7: condensing agent: organic base = 1: (0.5-1.5): (1-3): (1-3); the reaction temperature is 20-50℃ and the time is 8-16h.

8. A method for preparing a platinum (IV) prodrug with an NF-κB inhibitor as a ligand as described in claim 1, characterized in that, Includes the following steps; (1) Compound 3 was reacted with N-Boc-bromopropylamine to synthesize compound 11 under inorganic base catalysis; (2) Compound 11 was reacted with 4N hydrochloric acid / methanol solution to synthesize compound 13; (3) React compound 13 with succinic anhydride or glutaric anhydride to synthesize compound 16 or 17; (4) React compound 16 or 17 with compound 7 under the action of a condensing agent and an organic base to obtain compound 20 or 21; The synthesis route is shown below:

9. The preparation method according to claim 8, characterized in that: The inorganic base mentioned in step (1) is K2CO3; in molar amounts, compound 3: N-Boc-bromopropylamine: inorganic base = 1:(1~2):(1~5); the reaction temperature is 40~60℃ and the time is 8~16h; And / or, in step (2), the molar ratio of compound 11:4N hydrochloric acid / methanol solution is 1:(0.5~1.5); the reaction temperature is 20~30℃ and the time is 8~16h; And / or, in step (3), in molar amounts, compound 13: succinic anhydride or glutaric anhydride = 1:(2-5); the reaction temperature is 50-70℃ and the time is 8-16h; And / or, in step (4), in molar amounts, compound 16 or 17: compound 7: condensing agent: organic base = 1: (0.5-1.5): (1-3): (1-3); the reaction temperature is 20-50℃ and the time is 8-16h.

10. The use of a platinum (IV) prodrug with an NF-κB inhibitor as a ligand as described in claim 1 in the preparation of an anti-lung cancer drug.

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