Preparation method of beta-phenylethylamine-containing steroid A-pyrazinamide compound and application of beta-phenylethylamine-containing steroid A-pyrazinamide compound in resisting prostatic cancer

Abstract

The invention discloses a steroid A-pyrazinamide compound containing beta-phenylethylamine as well as a preparation method and application of the steroid A-pyrazinamide compound. The structural general formula of the compound is shown as a formula (I). The steroid A-pyrazinamide compound containing beta-phenylethylamine provided by the invention has a remarkable inhibitory activity on high metastatic prostate cancer C4-2B. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a synthesis method of a class of steroidal A-fused pyrazine amide compounds of β-phenethylamine and their application in the preparation of anti-prostate cancer drugs. Background Art

[0002] Malignant tumors seriously threaten human health. The common drug for metastatic prostate cancer patients is castration therapy, but almost all patients eventually progress to metastatic castration-resistant prostate cancer after castration therapy. The 5-year survival rate of such patients is only about 30%. Some steroidal derivatives have been developed as commercial anti-cancer drugs, but they have poor water solubility, low bioavailability, large dosage (1 g / day), and cause serious adverse reactions such as severe edema and hypertension, which will cause irreversible damage to the liver and kidneys, and obvious drug resistance has also emerged. From enzalutamide to abiraterone acetate, "drug resistance" has promoted a series of drug "revolutions". The research and development of drugs for anti-prostate cancer metastasis is imminent. Summary of the Invention

[0003] The inventors have discovered the efficient preparation of a class of steroidal A-fused pyrazine amide compounds containing β-phenethylamine and have significant inhibitory activity against highly metastatic prostate cancer cells C4-2B. The structural general formula of the compound is shown in formula (Ι):

[0004]

[0005] Wherein:

[0006] R is a fluorine atom, a chlorine atom, a bromine atom, a methoxy group, a hydroxyl group, 3,4-dimethoxy, 2,4-dimethoxy, 3,4,5-trimethoxy and 2,4,6-trimethoxy.

[0007] Specifically, the structure of the compound of the present invention selects one of the following structural formulas:

[0008]

[0009]

[0010] The present invention also provides pharmaceutically acceptable salts or solvents of the above compounds.

[0011] The present invention also provides a preparation method of the above compounds. For this purpose, the preparation method provided by the present invention includes:

[0012] 1) Using progesterone as a raw material, successively carry out carbonyl ethylene glycol protection, introduction of 4,4-dimethyl, and oxidation at the 2-position to obtain an important intermediate L4;

[0013]

[0014] Reagents and conditions: i. EG, (EtO)4Si, ρ-TSA (catalyst), room temperature, 5h; ii. t-BuOH, t-BuOK, 1h, then add MeI; iii.

[0015] t-BuOK,t-BuOH,THF,O2.

[0016] 2) Methyl esterification of diaminopropionic acid L5 to obtain L6, which is then reacted with L4 to obtain the key intermediate L7 of steroid A pyrazine carboxylate before functional group derivatization;

[0017]

[0018] Reagents and conditions: iv. SOCl2, MeOH, 95%; v. MeOH, ρ-TSA, reflux, 90%.

[0019] 3) Based on L7, the final amide compound L8 is obtained by reacting with substituted β-phenylethylamine, and then the final target product L9 is obtained by removing the glycol protection and reducing the branched carbonyl group. The reaction process is shown in the following formula:

[0020]

[0021] Reagents and reaction conditions: vi: (a) THF:MeOH:H2O (v / v, 5:3:2), NaOH, 95%; (b) amine, HATU, DCM, reflux. vii. (a) 1M HCl, 77%-90%; (b) NaBH4, dry MeOH / THF (v / v, 2:1), 0℃, 80%-90%.

[0022] At the same time, the present invention provides one of the intermediates for preparing the compound of the present invention, and the structural formula of the intermediate is shown in formula (II):

[0023]

[0024] The steroidal A-pyrazinamide compound containing beta-phenylethylamine provided by the present invention has significant inhibitory activity on highly metastatic prostate cells C4-2B, which is significantly higher than the activity against other tumor cells.

[0025] The advantage of the preparation method of the present invention is that the preparation process is more efficient. By first obtaining the key intermediate L7 of steroid A pyrazine carboxylate and then derivatizing the functional group, compared with the inventor's previous patent introducing the functional group synthesis path from scratch (CN201911234480.7), it greatly saves the workload and improves the yield of the target product. Secondly, it is more selective for prostate cancer subtype C4-2B. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1: 1H NMR spectrum of compound L9a (R = o-F). 1 1H NMR spectrum. Figure 2 : 1H NMR spectrum of compound L9b (R = m-F). 1 1H NMR spectrum. Figure 3 : 1H NMR spectrum of compound L9c (R = p-F). 1 1H NMR spectrum. Figure 4 : 1H NMR spectrum of compound L9d (R = o-Cl). 1 1H NMR spectrum. Figure 5 : 1H NMR spectrum of compound L9e (R = m-Cl). 1 1H NMR spectrum. Figure 6 : 1H NMR spectrum of compound L9f (R = p-Cl). 1 1H NMR spectrum. Figure 7 : 1H NMR spectrum of compound L9g (R = o-Br). 1 1H NMR spectrum. Figure 8 : 1H NMR spectrum of compound L9h (R = m-Br). 1 1H NMR spectrum. Figure 9 : 1H NMR spectrum of compound L9i (R = p-Br). 1 1H NMR spectrum. Figure 10 : 1H NMR spectrum of compound L9j (R = o-OMe). 1 1H NMR spectrum. Figure 11 : 1H NMR spectrum of compound L9k (R = m-OMe). 1 1H NMR spectrum. Figure 12 : 1H NMR spectrum of compound L9l (R = p-OMe). 1 1H NMR spectrum. Figure 13 : 1H NMR spectrum of compound L9m (R = OH). 1 1H NMR spectrum. Figure 14 : 1H NMR spectrum of compound L9n (R = 3,4-OMe). 1 1H NMR spectrum. Figure 15 : 1H NMR spectrum of compound L9o (R = 2,4-OMe). 1 1H NMR spectrum. Figure 16 : 1H NMR spectrum of compound L9p (R = 3,4,5-OMe). 1 1H NMR spectrum. Figure 17: of compound L9q (R = 2,4,6-OMe) 1 1H NMR spectrum. Detailed implementation mode

[0027] The following specifically illustrates various aspects and features of the present invention through examples of synthesizing steroid A-pyrazinecarboxamide compounds containing β-phenethylamine. Those skilled in the art should understand that these examples are only for illustrative purposes and do not limit the scope of the present invention. The protection scope of the present invention is only limited by the claims. Without departing from the scope of the claims, those skilled in the art can make various modifications and improvements to various aspects of the invention, and these modifications and improvements also belong to the protection scope of the present invention.

[0028] In addition, it should be noted that unless otherwise specified, various materials and reagents used in the following examples are common materials and reagents in the art and can be obtained through conventional commercial channels; the intermediates used can be prepared by conventional methods; the methods used are all conventional methods well-known to those skilled in the art.

[0029] Example:

[0030] The synthesis of the compounds described in the present invention can adopt the following methods:

[0031] 1) Methylate diaminopropionic acid L5 to obtain L6, and then react with L4 to obtain the key intermediate L7 of steroid A-pyrazinecarboxylate before functional group derivation;

[0032] 2) On the basis of L7, react with β-phenethylamine with different substitutions to form the final amide compound, and then simply remove the ethylene glycol protection and reduce the branched-chain carbonyl to obtain the final target product L9. The specific synthesis examples are as follows:

[0033] (1) The synthesis of the related intermediate L7 can refer to the following methods (the substituents used are shown in Table 1):

[0034]

[0035] i. EG, (EtO)4Si, ρ-TSA (cat.), r.t., overnight, 70%; ii. (a) t-BuOH, t-BuOK, 0.5 - 1 h, (b) MeI, 60%; iii. t-BuOK, t-BuOH, THF, O2, 45 °C, 40 min, 95%.

[0036] iv. (a) MeOH, 0 °C, SOCl2; (b) reflux, 4 h, 99%; v. (a) dry-MeOH, TEA, ρ-TSA, reflux, 85%.

[0037] L2: Dissolve 1.00 g of progesterone L1 (3.2 mmol) in 20 mL of ethylene glycol solution, then add 1.5 mL of tetraethyl orthosilicate (6.4 mmol) and a catalytic amount of p-toluenesulfonic acid, and stir overnight at room temperature. After the reaction is complete (monitored by thin-layer chromatography), back-extract with EtOAc (25×3 mL) and saturated sodium bicarbonate solution. The organic layer is dried over anhydrous Na2SO4 and concentrated. Purify by silica gel column chromatography (PE:EA / 3:1, visualized with sulfuric acid-ethanol), obtaining white solid L2 with a yield of 70%. 1 1H NMR (400 MHz, CDCl3) δ 5.70 - 5.66 (m, 1H, 4H), 3.93 (m, J = 25.1, 7.0 Hz, 2H,), 3.84 (h, J = 7.2 Hz, 2H), 2.40 - 2.20 (m, 4H), 2.06 - 1.95 (m, 2H), 1.84 - 1.74 (m, 2H), 1.72 - 1.60 (m, 4H), 1.55 - 1.46 (m, 2H), 1.44 - 1.37 (m, 1H), 1.25 (s, 3H), 1.19 - 1.16 (m, 1H), 1.15 (s, 3H), 1.14 - 1.09 (m, 1H), 1.04 - 0.96 (m, 2H), 0.89 (m, J = 11.6, 4.3 Hz, 1H), 0.77 (s, 3H); 13 13C NMR (125 MHz, CDCl3) δ 199.65, 171.61, 123.81, 111.81, 65.23, 63.24, 58.15, 55.76, 53.80, 41.86, 39.31, 38.64, 35.72, 35.12, 34.02, 32.96, 31.96, 24.60, 23.77, 22.95, 20.85, 17.42, 12.98.

[0038] L3: Dissolve compound L2 (4.7 mmol) in dry t-BuOH (20 mL) solution, then add t-BuOK (2.1 g, 19.0 mmol), protect with nitrogen, stir at room temperature for 0.5 - 1 h. After the solid dissolves, slowly add MeI (3.0 mL, 47.0 mmol) with a syringe. React overnight at room temperature. When the reaction solution becomes milky white (monitored by thin-layer chromatography, visualized with sulfuric acid-ethanol,

[0039] R f = 0.6, PE:EA / 10:1), place it in ice water without adjusting the pH. Extract with EtOAc (25×3 mL) and saturated NaCl solution (30×2 mL) to remove the residual t-BuOH. Combine the organic layers, dry over anhydrous Na2SO4 and concentrate

[21] A white solid compound L3 was obtained with a yield of 60%. 1 HNMR (400 MHz, CDCl3) δ 5.53 (dd, J = 5.4, 2.5 Hz, 1H, H6), 4.02

[0040] -3.91 (m, 2H), 3.86 (m, J = 7.3, 6.7 Hz, 2H), 2.57 - 2.40 (m, 2H, H2), 2.12 - 2.05 (m, 2H, H7), 2.03 - 1.97 (m, 1H), 1.79 (dd, J = 10.8, 8.2 Hz, 1H), 1.75 - 1.69 (m, 2H), 1.67 - 1.57 (m, 3H), 1.57

[0041] -1.46 (m, 2H), 1.44 - 1.37 (m, 1H), 1.28 (s, 3H), 1.21 (s, 6H), 1.19 - 1.08 (m, 2H), 1.08 - 0.98 (m, 2H), 0.83 (s, 3H,), 0.76 (s, 3H); 13 C NMR (125 MHz, CDCl3) δ 216.89, 149.85, 119.89, 111.91, 65.23, 63.23, 58.20, 56.66, 48.91, 48.68, 41.85, 39.42, 37.11, 33.74, 32.10, 31.65, 30.75, 30.27, 27.26, 24.61, 23.76, 23.02, 21.10, 19.37, 12.94.

[0042] L4: Compound L3 (10.00 mmol) was dissolved in a 50 mL mixed solution of dry t-BuOH / THF (20:1, v / v), and then 2.5 g of t-BuOK (25.00 mmol) was added. The reaction was carried out at 45 °C for 40 min (while passing O2)

[22] , monitored by thin-layer chromatography (PE:EA / 10:1, developed with H2SO4-EtOH), back-extracted with EtOAc and saturated NaCl solution, dried over anhydrous Na2SO4 and concentrated to obtain product L4, which is a pale yellow solid with a yield of 95%. 11H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H, OH), 6.03 (s, 1H, H1), 5.59 (d, J = 4.6 Hz, 1H, H6), 3.97 - 3.87 (m, 2H), 3.82 (m, J = 17.7, 7.0 Hz, 2H), 2.16 - 2.05 (m, 2H, H7), 1.77 - 1.50 (m, 8H), 1.32 (s, 3H), 1.27 (s, 3H), 1.23 (s, 6H), 1.20 - 1.02 (m, 4H), 0.78 (s, 3H); 13 13C NMR (125 MHz, CDCl3) δ 199.40, 143.84, 142.83, 123.11, 122.10, 111.86, 65.19, 63.22, 58.00, 57.06, 48.23, 47.25, 41.75, 39.19, 38.47, 32.08, 31.43, 30.55, 24.59, 23.97, 23.62, 23.20, 23.01, 20.77, 13.01.

[0043] L6: Dissolve 2,3-diaminopropionic acid L5 (0.608 g, 4.8 mmol) in 20 mL of ethanol solution, cool to 0 °C, and then add thionyl chloride (1.4 mL, 19.2 mmol, 4 eq.) dropwise within 10 minutes. Heat the reaction mixture under reflux for 4 h. Then remove the excess thionyl chloride and solvent under vacuum.

[23] . Compound L6 (0.50 g, 4.26 mmol, 99%) was obtained as a colorless solid without further purification.

[0044] L7: To a solution of compound L6 (200 mg, 0.5 mmol, 1.1 eq.) in anhydrous ethanol (10 mL), add L4 (400 mg, 1 mmol, 2.2 eq.), 1 mL of Et3N solution, a catalytic amount of ρ-TSA, and reflux the mixture at 75 °C for 8 h. After the reaction is complete, add an appropriate volume of silica gel for concentration and sample mixing, and purify by silica gel column chromatography (PE / EtOAc, 5:1, v / v) to obtain white solid L7 with a yield of 85%. 11H NMR (400 MHz, Chloroform-d) δ 8.96 (s, 1H), 5.81 (dd, J = 5.3, 2.4 Hz, 1H), 4.46 (q, J = 7.1 Hz, 2H), 4.05 - 3.89 (m, 4H), 2.93 (dd, J = 200.9, 15.9 Hz, 2H), 2.04 (s, 1H), 1.68 (s, 3H), 1.43 (t, J = 7.1 Hz, 3H), 1.34 (s, 3H), 1.31 (s, 3H), 0.80 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 164.73, 159.57, 154.71, 148.50, 142.45, 141.67, 120.92, 111.90, 65.19, 63.21, 61.72, 58.17, 56.68, 48.85, 44.03, 41.79, 41.59, 39.34, 37.73, 33.69, 31.75, 31.46, 30.94, 24.57, 23.74, 23.00, 20.90, 20.38, 14.29, 12.89.

[0045] (II) Synthetic reference method for the final compound (substituents used are shown in Table 1):

[0046] Synthetic route of compounds L9a - q:

[0047]

[0048] vi. (a) MeOH / 10% NaOH (v / v, 4:1), 15 min; (b) dry-EtOH, Et3N, HATU, β-PEA, reflux, 12 h, 81%; (c) 6M HCl, NaHCO3, 98%.

[0049] vii. NaBH4, MeOH, THF, 99%.

[0050] Table 1

[0051]

[0052] Taking the synthesis of compound L9a as an example:

[0053] L8a: Methanol and 10% aqueous NaOH solution (4:1, v:v) were added to L7 and reacted for 15 min to obtain a hydrolysis product. Then the product was dissolved in 10 mL of anhydrous ethanol solution, 1 mL of Et3N and a certain amount of HATU were added successively, and the mixture was stirred at room temperature for 30 min for activation. Then β-phenethylamine (139.70 mg, 1.5 eq.) was added, and the mixture was refluxed under condensation at 75 °C for 12 h. After detection by TLC plate and completion of the reaction, the temperature was lowered to room temperature, and the mixture was extracted with saturated brine and DCM (25 mL × 3). The lower organic phase was taken, dried with anhydrous Na2SO4 and concentrated. The crude product was obtained by flash column chromatography (dichloromethane:methanol = 60:1). It was dissolved in DCM / MT (5 mL, 1:1, v / v), 3 drops of 6 M hydrochloric acid were added to the solution, and the mixture was stirred for 5 - 10 min. After completion of the reaction, the residual hydrochloric acid in the solution was neutralized with solid NaHCO3, filtered, the filtrate was taken, an appropriate volume of silica gel was added for concentration and sample mixing, and purification was carried out by silica gel column chromatography (PE / EtOAc, 5:1, v / v) to obtain white solid L8a with a yield of 81%.

[0054] L9a: The synthesized L8 (0.1 mmol) was dissolved in dry MT / THF (2 mL, 5:1, v / v) solution, and sodium borohydride (100 mg) was added in batches under ice bath conditions and stirred for 10 min. After completion of the reaction, methanol was added for dissolution, 2 drops of hydrochloric acid were added for quenching, and the THF solution was removed by rotary evaporation. Purification was carried out by silica gel column chromatography (PE / EA, 1:1) to obtain the reduction product, which was white solid L9a with a yield of 98%. 1 HNMR(400MHz,Chloroform-d)δ9.08(s,1H),7.90(t,J=6.1Hz,1H),7.28-7.15(m,2H),7.12-7.01(m,2H),5.81(dd,J=5.3,2.5Hz,1H),3.20(d,J=15.9Hz,1H),2.67(d,J=15.9Hz,1H),1.57(s,3H),1.25(s,3H),1.17(d,J=6.1Hz,3H),0.79(s,3H),0.77(s,3H). 1313C NMR (100 MHz, CDCl3) δ 163.71, 160.07, 157.48, 154.46, 148.28, 142.17, 140.41, 131.27, 131.22, 128.53, 128.45, 124.33, 124.30, 121.06, 115.58, 115.36, 70.47, 58.33, 56.26, 48.84, 43.83, 42.30, 41.20, 39.76, 39.40, 37.80, 33.70, 31.92, 31.64, 31.27, 29.73, 29.73, 29.15, 25.69, 24.50, 23.80, 20.97, 20.49, 12.44.

[0055] The synthetic methods of L8b-q and L9b-q are the same as above, and the specific NMR data are as follows:

[0056] L9b: 1 1H NMR (400 MHz, DMSO-d6) δ 8.87 (s, 1H), 8.71 - 8.37 (m, 1H), 7.39 - 7.24 (m, 1H), 7.15 - 7.07 (m, 2H), 7.07 - 6.99 (m, 2H), 5.89 - 5.79 (m, 1H), 1.62 (s, 3H), 1.23 (s, 3H), 1.01 (d, J = 6.1 Hz, 3H), 0.72 (s, 3H), 0.71 (s, 3H). 13 13C NMR (100 MHz, CDCl3) δ 163.72, 162.80, 149.23, 148.46, 141.69, 140.80, 130.14, 130.05, 124.46, 121.00, 115.85, 115.65, 113.58, 113.37, 70.59, 58.37, 56.30, 48.85, 43.63, 42.31, 41.74, 40.41, 39.78, 37.67, 35.74, 33.55, 31.91, 31.80, 31.25, 29.73, 29.73, 25.70, 24.48, 23.81, 21.03, 20.47, 12.46.

[0057] L9c: 11H NMR (400 MHz, Chloroform-d) δ 9.11 (s, 1H), 8.10 (t, J = 6.3 Hz, 1H), 7.34 (dd, J = 8.7, 5.3 Hz, 2H), 7.03 (t, J = 8.6 Hz, 2H), 5.79 (dd, J = 5.3, 2.6 Hz, 1H), 3.21 (d, J = 16.0 Hz, 1H), 2.67 (d, J = 15.9 Hz, 1H), 1.59 (s, 3H), 1.31 (s, 3H), 1.16 (d, J = 6.1 Hz, 3H), 0.77 (s, 3H), 0.76 (s, 3H). 13 13C NMR (100 MHz, CDCl3) δ 163.78, 157.73, 154.77, 148.17, 142.09, 140.60, 129.41, 129.33, 121.16, 115.71, 115.50, 70.34, 58.27, 56.26, 48.85, 43.79, 42.60, 42.31, 41.24, 39.76, 37.78, 33.81, 31.90, 31.67, 31.27, 29.71, 25.68, 24.49, 23.82, 20.99, 20.47, 12.4. 19 19F NMR (376 MHz, DMSO) δ -113.65.

[0058] L9d: 1 1H NMR (500 MHz, Chloroform-d) δ 9.06 (s, 1H), 7.90 (t, J = 6.1 Hz, 1H), 7.38 (dd, J = 6.5, 2.6 Hz, 1H), 7.30 (td, J = 6.1, 5.2, 2.6 Hz, 2H), 7.21 (td, J = 6.5, 6.0, 3.7 Hz, 3H), 5.81 (dd, J = 5.4, 2.6 Hz, 1H), 4.12 (q, J = 7.2 Hz, 1H), 3.91 - 3.61 (m, 4H), 3.45 - 3.38 (m, 1H), 3.24 (d, J = 15.9 Hz, 1H), 3.11 (t, J = 6.9 Hz, 3H), 2.93 (t, J = 7.0 Hz, 1H), 2.67 (d, J = 15.9 Hz, 1H), 2.36 - 2.16 (m, 3H), 1.57 (s, 3H), 1.32 (s, 3H), 1.19 (d, J = 6.0 Hz, 3H), 0.78 (s, 6H). 1313C NMR (101 MHz, CDCl3) δ 163.59, 157.72, 154.60, 148.13, 142.14, 140.86, 140.21, 134.47, 129.97, 128.99, 127.04, 126.86, 121.16, 70.14, 58.17, 56.24, 48.83, 43.65, 42.33, 41.17, 40.18, 39.77, 37.78, 35.51, 33.75, 31.91, 31.60, 31.28, 25.70, 24.51, 23.93, 21.01, 20.47, 12.39.

[0059] L9e: 1 1H NMR (400 MHz, Chloroform-d) δ 9.08 (d, J = 7.9 Hz, 1H), 7.87 (t, J = 6.3 Hz, 1H), 7.27 (dd, J = 13.7, 5.6 Hz, 4H), 6.29 - 5.50 (m, 1H), 3.74 (q, J = 7.0 Hz, 3H), 3.23 (d, J = 15.9 Hz, 1H), 2.67 (d, J = 15.9 Hz, 1H), 1.55 (s, 3H), 1.30 (s, 3H), 1.18 (d, J = 6.3 Hz, 3H), 0.78 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.59, 157.72, 154.60, 148.13, 142.14, 140.86, 140.21, 134.47, 129.97, 128.99, 127.04, 126.86, 121.16, 70.14, 58.17, 56.24, 48.83, 43.65, 42.33, 41.17, 40.18, 39.77, 37.78, 35.51, 33.75, 31.91, 31.60, 31.28, 25.70, 24.51, 23.93, 21.01, 20.47, 12.39.

[0060] L9f: 11H NMR (400 MHz, Chloroform-d) δ 9.07 (s, 1H), 7.93 - 7.59 (m, 1H), 7.31 - 7.28 (m, 2H), 7.22 - 7.18 (m, 2H), 5.81 (dd, J = 5.3, 2.5 Hz, 1H), 3.83 - 3.56 (m, 2H), 3.21 (d, J = 16.0 Hz, 1H), 2.93 (t, J = 6.9 Hz, 2H), 2.67 (d, J = 16.0 Hz, 1H), 2.30 - 2.14 (m, 1H), 1.77 - 1.65 (m, 1H), 1.54 (s, 3H), 1.29 (s, 2H), 1.17 (d, J = 6.1 Hz, 2H), 0.79 (s, 2H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.60, 157.58, 154.60, 148.15, 142.13, 140.33, 137.24, 132.49, 130.21, 128.84, 121.18, 70.33, 58.28, 56.26, 48.86, 43.75, 42.32, 41.17, 40.22, 39.77, 37.79, 35.18, 33.71, 31.91, 31.56, 31.28, 25.69, 24.50, 23.84, 20.99, 20.49, 12.41.

[0061] L9g: 1 1H NMR (400 MHz, Chloroform-d) δ 9.01 (s, 1H), 7.77 (t, J = 5.8 Hz, 1H), 7.17 - 7.08 (m, 2H), 6.91 - 6.74 (m, 2H), 5.82 - 5.67 (m, 1H), 3.77 (s, 3H), 3.12 (d, J = 16.0 Hz, 1H), 2.90 (t, J = 6.8 Hz, 2H), 2.59 (d, J = 16.0 Hz, 1H), 1.49 (s, 3H), 1.22 (d, J = 2.3 Hz, 3H), 1.10 (d, J = 6.1 Hz, 3H), 0.72 (s, 3H), 0.70 (s, 3H). 1313C NMR (101 MHz, CDCl3) δ 162.58, 156.57, 156.35, 153.19, 147.29, 141.40, 139.36, 129.61, 126.94, 126.05, 119.98, 119.59, 109.47, 69.38, 57.27, 55.22, 54.36, 47.80, 42.74, 41.26, 40.14, 38.72, 38.09, 36.75, 32.65, 30.88, 30.63, 30.24, 29.27, 28.68, 24.64, 23.46, 22.76, 19.92, 19.43, 11.38.

[0062] L9h: 1 1H NMR (400 MHz, Chloroform-d) δ 9.08 (s, 1H), 7.86 (t, J = 6.1 Hz, 1H), 7.24 (dd, J = 7.3, 1.5 Hz, 1H), 6.86 (dt, J = 7.4, 1.3 Hz, 1H), 6.80 (dd, J = 7.3, 1.1 Hz, 2H), 5.80 (dd, J = 5.3, 2.6 Hz, 1H), 3.79 (s, 3H), 3.20 (d, J = 16.0 Hz, 1H), 2.92 (t, J = 6.9 Hz, 2H), 2.66 (d, J = 15.9 Hz, 1H), 1.53 (s, 2H), 1.29 (s, 3H), 1.17 (d, J = 6.1 Hz, 3H), 0.79 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.58, 159.90, 157.49, 154.44, 148.29, 142.24, 140.38, 129.73, 121.15, 121.05, 114.59, 111.93, 70.42, 58.33, 56.27, 55.14, 48.86, 43.80, 42.31, 41.17, 40.31, 39.77, 37.80, 35.83, 33.69, 31.91, 31.58, 31.28, 25.68, 24.50, 23.79, 20.98, 20.47, 12.42.

[0063] L9i: 11H NMR (400 MHz, Chloroform-d) δ 9.08 (s, 1H), 7.82 (t, J = 6.1 Hz, 1H), 7.53 - 7.35 (m, 2H), 7.20 - 7.06 (m, 2H), 5.81 (dd, J = 5.3, 2.5 Hz, 1H), 3.88 - 3.51 (m, 2H), 3.20 (d, J = 16.0 Hz, 1H), 2.91 (t, J = 6.9 Hz, 2H), 2.67 (d, J = 16.0 Hz, 1H), 1.54 (s, 3H), 1.29 (s, 3H), 1.17 (d, J = 6.1 Hz, 3H), 0.79 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.62, 157.51, 154.60, 148.18, 142.09, 140.41, 137.77, 131.81, 131.81, 130.60, 130.60, 121.15, 120.52, 70.46, 58.35, 56.27, 48.86, 43.83, 42.31, 41.18, 40.14, 39.77, 37.80, 35.25, 33.72, 31.91, 31.57, 31.28, 25.68, 24.49, 23.78, 20.98, 20.49, 12.42.

[0064] L9j: 1 1H NMR (400 MHz, DMSO-d6) δ 8.83 - 8.68 (m, 1H), 7.60 (dd, J = 8.0, 1.3 Hz, 2H), 7.38 (dd, J = 7.6, 1.8 Hz, 1H), 7.31 (td, J = 7.4, 1.3 Hz, 2H), 7.24 - 7.03 (m, 1H), 6.04 - 5.74 (m, 1H), 4.16 (d, J = 5.6 Hz, 1H), 1.63 (s, 3H), 1.33 (s, 3H), 1.23 (s, 4H), 1.01 (d, J = 6.0 Hz, 3H), 0.72 (s, 3H), 0.71 (s, 3H). 1313C NMR (101 MHz, DMSO) δ 163.66, 157.45, 154.03, 148.69, 143.13, 140.28, 138.93, 133.02, 131.59, 128.98, 128.31, 124.39, 120.89, 68.77, 58.06, 56.37, 48.82, 43.62, 42.37, 41.46, 37.69, 35.72, 33.71, 32.11, 31.97, 31.33, 29.52, 25.83, 24.64, 24.29, 22.58, 20.91, 20.74, 12.45.

[0065] L9k: 1 1H NMR (400 MHz, Chloroform-d) δ 9.07 (s, 1H), 7.86 (t, J = 6.2 Hz, 1H), 7.45 - 7.39 (m, 1H), 7.38 (td, J = 3.8, 1.8 Hz, 1H), 7.23 - 7.17 (m, 2H), 5.81 (dd, J = 5.3, 2.5 Hz, 1H), 3.23 (d, J = 16.0 Hz, 1H), 2.92 (t, J = 6.9 Hz, 2H), 2.67 (d, J = 16.0 Hz, 1H), 1.56 (s, 3H), 1.30 (s, 3H), 1.18 (d, J = 6.1 Hz, 3H), 0.78 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.59, 157.73, 154.62, 148.13, 142.13, 141.17, 140.22, 131.91, 130.27, 129.80, 127.52, 122.75, 121.17, 70.16, 58.17, 56.24, 48.82, 43.65, 42.33, 41.19, 40.21, 39.78, 37.79, 35.50, 33.79, 31.92, 31.65, 31.28, 25.70, 24.52, 23.95, 21.00, 20.49, 12.40.

[0066] L9l: 11H NMR (400 MHz, Chloroform-d) δ 9.07 (d, J = 4.8 Hz, 1H), 7.83 (t, J = 6.1 Hz, 1H), 7.53 - 7.37 (m, 2H), 7.15 (t, J = 6.4 Hz, 2H), 5.81 (d, J = 6.0 Hz, 1H), 3.74 (dq, J = 13.1, 6.8, 6.1 Hz, 3H), 3.22 (dd, J = 15.9, 4.9 Hz, 1H), 2.91 (t, J = 6.8 Hz, 2H), 2.67 (dd, J = 15.9, 5.0 Hz, 1H), 2.21 (dt, J = 12.0, 5.4 Hz, 1H), 1.55 (s, 3H), 1.30 (s, 3H), 1.17 (d, J = 6.0 Hz, 3H), 0.79 (s, 3H), 0.77 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 163.60, 157.61, 154.60, 148.15, 142.13, 140.29, 137.75, 131.80, 131.80, 130.59, 130.59, 121.18, 120.51, 70.28, 58.27, 56.27, 48.88, 43.73, 42.32, 41.18, 40.15, 39.77, 37.79, 35.23, 33.71, 31.91, 31.56, 31.29, 25.69, 24.50, 23.85, 21.00, 20.49, 12.40.

[0067] L9m: 1 1H NMR (400 MHz, Chloroform-d) δ 9.23 (s, 1H), 7.90 (t, J = 5.9 Hz, 1H), 6.97 - 6.57 (m, 3H), 3.87 (d, J = 7.0 Hz, 6H), 2.16 (s, 3H), 1.63 (s, 3H), 1.34 (s, 3H), 0.77 (s, 3H), 0.67 (s, 3H). 1313C NMR (101 MHz, CDCl3) δ 209.49, 165.98, 163.62, 148.94, 148.90, 148.41, 147.65, 141.86, 134.23, 131.47, 129.50, 120.90, 120.70, 111.94, 111.34, 67.77, 63.59, 56.96, 55.93, 55.85, 48.72, 43.97, 43.66, 41.75, 40.87, 38.90, 38.74, 37.66, 35.55, 33.55, 31.82, 31.75, 31.60, 31.37, 30.57, 29.00, 23.98, 22.99, 20.44, 14.08, 13.32, 11.12.

[0068] L9n: 1H NMR (400 MHz, Chloroform-d) δ 9.08 (s, 1H), 7.82 (s, 1H), 7.27 (s, 1H), 7.09 (d, J = 8.2 Hz, 1H), 6.47 (d, J = 2.2 Hz, 1H), 6.43 (dd, J = 8.3, 2.3 Hz, 1H), 5.80 (d, J = 2.8 Hz, 1H), 3.80 (d, J = 6.7 Hz, 6H), 3.68 (d, J = 6.4 Hz, 2H), 3.17 (s, 1H), 2.90 (t, J = 6.9 Hz, 2H), 2.66 (d, J = 15.7 Hz, 1H), 2.19 (dd, J = 14.5, 9.9 Hz, 3H), 1.57 (s, 3H), 1.37 (s, 1H), 1.33 (s, 1H), 1.30 (s, 3H), 0.79 (s, 3H), 0.77 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 163.62, 159.79, 158.52, 157.32, 154.21, 148.36, 142.47, 140.45, 130.89, 121.02, 119.43, 104.07, 98.77, 70.47, 58.35, 56.27, 55.43, 55.38, 48.86, 43.82, 42.30, 41.18, 39.76, 39.29, 33.70, 31.92, 31.69, 31.28, 25.68, 24.49, 23.77, 20.96, 20.47, 12.42.

[0069] L9o: 11H NMR (400 MHz, Chloroform-d) δ 7.83 (s, 1H), 6.13 (s, 2H), 3.80 (s, 6H), 3.78 (s, 3H), 3.63 (q, J = 6.2 Hz, 2H), 2.68 (d, J = 15.8 Hz, 1H), 2.15 (s, 6H), 2.05 (s, 3H), 1.61 (s, 3H), 1.55 (d, J = 8.1 Hz, 2H), 1.33 (s, 3H), 1.28 (s, 1H), 1.26 (s, 1H), 0.77 (s, 3H), 0.65 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 209.44, 163.83, 159.86, 159.13, 157.07, 157.07, 153.63, 148.35, 142.77, 140.54, 120.85, 107.63, 90.62, 63.60, 56.96, 56.96, 55.83, 55.32, 48.69, 43.97, 43.82, 41.15, 39.41, 38.75, 37.74, 33.62, 31.74, 31.58, 31.39, 24.41, 24.41, 22.81, 21.87, 21.09, 20.42, 13.28.

[0070] L9p: 1 1H NMR (400 MHz, Chloroform-d) δ 9.10 (s, 1H), 7.94 (t, J = 6.0 Hz, 1H), 7.16 - 6.95 (m, 2H), 6.93 - 6.67 (m, 2H), 5.80 (dd, J = 5.4, 2.5 Hz, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.71 (q, J = 6.7 Hz, 2H), 3.20 (d, J = 15.9 Hz, 1H), 2.87 (t, J = 6.9 Hz, 2H), 2.68 (d, J = 15.9 Hz, 1H), 2.16 (s, 3H), 2.05 (s, 3H), 1.57 (s, 3H), 0.76 (s, 3H), 0.65 (s, 3H). 1313C NMR (101 MHz, CDCl3) δ 163.63, 157.54, 154.57, 154.48, 148.20, 142.22, 140.38, 130.64, 130.00, 130.00, 121.11, 115.63, 115.63, 70.52, 58.34, 56.26, 48.82, 43.79, 42.30, 41.20, 40.64, 39.76, 37.79, 34.90, 33.74, 31.64, 31.53, 31.27, 30.14, 25.69, 23.77, 20.96, 20.50, 12.45.

[0071] L9q: 1 1H NMR (400 MHz, Chloroform-d) δ 8.10 (s, 1H), 7.83 (s, 1H), 6.48 (s, 2H), 5.82 - 5.78 (m, 1H), 3.86 (s, 3H), 3.83 (s, 6H), 3.76 (s, 2H), 2.89 - 2.62 (m, 2H), 2.58 (t, J = 8.9 Hz, 1H), 2.15 (s, 3H), 1.89 (s, 1H), 1.74 (s, 1H), 1.61 (s, 2H), 1.53 (s, 3H), 1.46 (d, J = 7.8 Hz, 1H), 1.33 (s, 1H), 1.28 (s, 3H), 0.76 (s, 3H), 0.66 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 209.40, 163.61, 157.38, 154.32, 153.37, 142.23, 140.53, 134.43, 134.23, 129.49, 121.06, 105.62, 104.70, 67.77, 63.61, 60.88, 56.96, 56.17, 56.07, 48.70, 43.97, 43.86, 41.16, 40.46, 38.90, 38.76, 37.76, 36.12, 33.74, 31.72, 31.58, 31.38, 30.57, 28.99, 24.41, 23.99, 22.98, 22.84, 21.11, 20.42, 14.06, 13.30, 11.11.

[0072] In vitro anti-tumor experiments of some compounds of the present invention:

[0073] Human prostate cancer C4-2B cells were selected. The inhibition rate was tested by the CCK8 method, and each treatment was repeated six times. The specific method is as follows:

[0074] The prostate tumor cells C4-2B were seeded into a 96-well cell culture plate at a density of 6000 cells / well, with 100 μL of cell suspension added to each well. The cell plate was placed in an incubator at 37 °C with 5% CO2 and cultured for 24 hours. Then, 100 μL of the compound working solution at a concentration of 20 μM was added to each well in the experimental group, and the plate was incubated in the dark at 37 °C with 5% CO2 for 72 hours. After the incubation, the medium was replaced with fresh medium containing 10% CCK8, and the plate was incubated in the incubator at 37 °C with 5% CO2 for 1 - 3 hours. The absorbance at 450 nm was measured using a microplate reader, and the experimental results of the tumor cell inhibition rate are shown in Table 2. According to the method described in the CCK8 kit instruction manual, the calculation formula for the cell inhibition rate is as follows:

[0075] Cell inhibition rate (%) = 1 - [A (drug added) - A (blank)] / [A (0 drug added) - A (blank)] × 100

[0076] Note: A (drug added): absorbance of the well with cells, CCK8 solution, and drug solution; A (0 drug added): absorbance of the well with cells, CCK8 solution, and 0.5% DMSO instead of the drug solution; A (blank): absorbance of the well with medium and CCK8 solution but without cells.

[0077] Table 2 Inhibition rates of compounds 9a - q on highly metastatic prostate cancer cells C4-2B

[0078]

[0079] The above experimental results show that the compounds of the present invention, especially compounds L9I and L9, can significantly inhibit the proliferation of prostate tumor cells at a low concentration (10 μM).

Claims

1. A compound and a pharmaceutically acceptable salt thereof, wherein the structure of the compound is as shown in formula (I): in: R is a fluorine atom, a chlorine atom, a bromine atom, a methoxy group, a hydroxy group, a 3,4-dimethoxy group, a 2,4-dimethoxy group, a 3,4,5-trimethoxy group, and a 2,4,6-trimethoxy group.

2. The compound and pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from one of the following structural formulas:

3. A method for preparing the compound according to claim 1 or 2 and a pharmaceutically acceptable salt thereof, the method comprising the following steps: 1) Using progesterone as the raw material, carbonyl glycol protection, 4,4-dimethylation, and 2-position oxidation were performed in sequence to obtain the important intermediate L4; 2) Methyl esterification of diaminopropionic acid L5 to obtain L6, which is then reacted with L4 to obtain the key intermediate L7 of steroid A pyrazine carboxylate before functional group derivatization; 3) Based on L7, the final amide compound L8 is obtained by reacting with substituted β-phenylethylamine, and then the final target product L9 is obtained by removing the glycol protection and reducing the branched carbonyl group. The reaction process is shown in the following formula:

4. An intermediate, the structural formula of which is shown in formula (II):

5. Use of the intermediate according to claim 4 for preparing the compound according to claim 1 or 2.

6. Use of the compound according to claim 1 or 2 and a pharmaceutically acceptable salt thereof in the preparation of a drug for treating prostate cancer.

7. The use according to claim 6, wherein the prostate cancer is highly metastatic prostate cancer.

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