Dihydroartemisinin-steroid conjugate and preparation method and application thereof

Abstract

Described is a dihydroartemisinin-steroid conjugate of formula (I), or a pharmaceutically acceptable salt thereof, where position 10 of dihydroartemisinin is linked to the steroid through a linker X. This application further provides a preparation method of the dihydroartemisinin-steroid conjugate and an application of the dihydroartemisinin-steroid conjugate in the preparation of a drug for treating cancer. The dihydroartemisinin-steroid conjugate of the invention exhibits potent inhibitory activity against various tumor cells and low cytotoxicity, moreover, the conjugate is capable of penetrating the blood-brain barrier, having a broad application prospect. ##STR00001##

Claims

1. A dihydroartemisinin-steroid conjugate or a pharmaceutically-acceptable salt thereof, the dihydroartemisinin-steroid conjugate being selected from the group consisting of: ##STR00028## ##STR00029##

2. A pharmaceutical composition comprising the dihydroartemisinin-steroid conjugate of claim I or a pharmaceutically-acceptable salt thereof and a pharmaceutically-acceptable carrier.

3. A method of preparing the dihydroartemisinin-steroid conjugate of claim 1, comprising: linking a hydroxyl group of dihydroartemisinin at position 10 with a hydroxyl group of the steroid compound at position 3 to form an ether bond so as to obtain the synthesizing the dihydroartemisinin-steroid conjugate.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The invention will be further illustrated with reference to the accompanying drawings and embodiments.

(2) FIG. 1 is a tumor growth curve revealing an anti-cancer effect of JZ-2 prepared in Example 15 of the invention on the xenograft model of human lung cancer cell A549; and

(3) FIG. 2 is an experimental animal body weight change curve revealing the anti-cancer effect of JZ-2 prepared in Example 15 of the invention on the xenograft model of human lung cancer cell A549.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Example 1

(4) Synthesis of JZ-01

(5) ##STR00016##

(6) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and cholic acid (817 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and then the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and then the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to obtain a crude product, which was purified by silica gel column chromatography to give a colorless oily compound JZ-01 (726 mg, yield 53.8%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 3:1-1:1.

(7) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.47 (s, 1H), 4.93 (d, J=3.3 Hz, 1H), 4.00 (s, 1H), 3.88 (s, 1H), 3.51 (t, J=11.0 Hz, 1H), 2.64-2.56 (m, 1H), 2.47-2.21 (m, 4H), 2.10-2.02 (m, 2H), 1.99-1.85 (m, 5H), 1.72 (m, 8H), 1.53 (m, 5H), 1.45 (m, 6H), 1.40-1.30 (m, 5H), 1.31-1.23 (m, 4H), 1.16 (m, 2H), 0.99 (m, 7H), 0.89 (m, 8H), 0.71 (s, 3H).

(8) .sup.13C NMR (100 MHz, CDCl.sub.3): 104.03, 100.14, 88.08, 81.29, 68.67, 52.65, 46.86, 46.51, 44.56, 42.21, 41.33, 39.32, 37.47, 36.51, 35.58, 35.29, 35.22, 34.87, 34.73, 31.53, 30.84, 30.68, 30.54, 30.31, 30.18, 30.35, 30.25, 29.72, 28.71, 27.58, 26.70, 26.30, 24.70, 23.22, 22.60, 20.41, 17.24, 13.08, 12.46.

Example 2

(9) Synthesis of JZ-02

(10) ##STR00017##

(11) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and ursodeoxycholic acid (785 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and then the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound JZ-02 (748 mg, yield 56.8%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 4:1-2:1.

(12) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.46 (s, 1H), 4.92 (d, J=3.3 Hz, 1H), 3.61 (m, 2H), 2.67-2.55 (m, 1H), 2.47-2.34 (m, 2H), 2.28 (ddd, J=15.8, 9.6, 6.5 Hz, 1H), 2.04 (m, 2H), 1.94-1.71 (m, 9H), 1.71-1.55 (m, 4H), 1.54-1.41 (m, 11H), 1.40-1.21 (m, 9H), 1.19-1.01 (m, 3H), 0.98-0.89 (m, 12H), 0.70 (s, 3H).

(13) .sup.13C NMR (100 MHz, CDCl.sub.3): 178.71, 104.07, 100.21, 88.08, 81.22, 71.41, 55.61, 54.92, 52.61, 44.51, 43.79, 42.37, 40.07, 39.19, 37.49, 36.99, 36.48, 35.21, 35.01, 34.71, 34.25, 33.60, 30.84, 30.82, 30.75, 28.85, 28.63, 26.90, 26.27, 24.70, 20.56, 23.53, 21.24, 20.41, 18.42, 13.16, 12.15.

Example 3

(14) Synthesis of JZ-03

(15) ##STR00018##

(16) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and hyodeoxycholic acid (785 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined to be completed by TLC, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was then extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound JZ-03 (653 mg, yield 49.5%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 8:1-1:1.

(17) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.41 (s, 1H), 4.96 (d, J=3.5 Hz, 1H), 4.14-4.02 (m, 1H), 3.64 (dt, J=15.5, 5.5 Hz, 1H), 2.68-2.58 (m, 1H), 2.35 (m, 3H), 2.12-1.95 (m, 2H), 1.95-1.56 (m, 12H), 1.56-1.21 (m, 16H), 1.12 (m, 6H), 1.02-0.89 (m, 10H), 0.86 (d, J=7.3 Hz, 3H), 0.65 (s, 3H).

(18) .sup.13C NMR (100 MHz, CDCl.sub.3): 179.13, 104.09, 98.78, 88.20, 81.19, 72.49, 71.73, 56.21, 55.93, 52.64, 47.88, 44.45, 42.88, 39.99, 37.55, 36.51, 36.00, 35.43, 35.34, 34.69, 34.51, 31.64, 30.24, 30.36, 30.44, 30.75, 30.77, 30.87, 29.67, 28.11, 26.26, 24.64, 24.22, 23.77, 20.75, 20.41, 18.25, 13.16, 12.02.

Example 4

(19) Synthesis of JZ-04

(20) ##STR00019##

(21) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and chenodeoxycholic acid (785 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined to be completed by TLC, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound JZ-04 (723 mg, yield 54.8%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 10:1-1:1 .sup.1H NMR (400 MHz, CDCl.sub.3): 5.46 (s, 1H), 4.93 (d, J=3.1 Hz, 1H), 3.88 (s, 1H), 3.48 (t, J=10.8 Hz, 1H), 2.65-2.55 (m, 1H), 2.46-2.33 (m, 2H), 2.27 (m, 1H), 1.87 (m, 16H), 1.52-1.13 (m, 21H), 0.95 (m, 14H), 0.66 (s, 3H).

(22) .sup.13C NMR (100 MHz, CDCl.sub.3): 179.74, 104.02, 100.40, 88.09, 81.30, 77.49, 68.70, 55.79, 52.63, 50.53, 44.59, 42.73, 41.33, 39.56, 39.33, 37.47, 36.51, 35.86, 35.39, 35.37, 35.23, 34.89, 34.72, 32.88, 30.94, 30.92, 30.79, 29.12, 28.18, 26.29, 24.70, 24.57, 23.70, 22.90, 20.64, 20.41, 18.27, 13.20, 11.78.

Example 5

(23) Synthesis of JZ-05

(24) ##STR00020##

(25) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and diosgenin (829 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a white solid compound JZ-05 (975 mg, yield 71.6%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 40:1-15:1.

(26) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.48 (s, 1H), 5.36 (d, J=4.9 Hz, 1H), 4.94 (d, J=3.4 Hz, 1H), 4.43 (dd, J=15.0, 7.5 Hz, 1H), 3.60 (dt, J=9.6, 4.7 Hz, 1H), 3.50 (dd, J=10.3, 3.5 Hz, 1H), 3.40 (t, J=10.9 Hz, 1H), 2.67-2.57 (m, 1H), 2.44-2.26 (m, 3H), 2.02 (m, 3H), 1.93-1.80 (m, 5H), 1.78-1.72 (m, 2H), 1.71-1.57 (m, 7H), 1.56-1.44 (m, 8H), 1.40-1.09 (m, 7H), 1.06-0.93 (m, 11H), 0.90 (d, J=7.3 Hz, 3H), 0.81 (m, 6H).

(27) .sup.13C NMR (100 MHz, CDCl.sub.3): 140.74, 121.39, 109.32, 104.05, 99.45, 88.09, 81.27, 80.87, 76.13, 66.87, 62.05, 56.46, 52.62, 50.01, 44.52, 41.61, 40.27, 39.78, 37.47, 36.94, 36.86, 36.49, 34.71, 32.07, 31.87, 31.42, 31.39, 30.76, 30.32, 28.81, 27.59, 26.28, 24.70, 24.52, 20.87, 20.42, 19.52, 17.18, 16.32, 14.57, 13.12.

Example 6

(28) Synthesis of JZ-06

(29) ##STR00021##

(30) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and ergosterol (793 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a white solid compound (895 mg, yield 67.5%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 30:1-15:1.

(31) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.56 (dd, J=5.6, 2.2 Hz, 1H), 5.49 (s, 1H), 5.39 (dt, J=5.3, 2.5 Hz, 1H), 5.33-5.14 (m, 2H), 4.97 (d, J=3.4 Hz, 1H), 3.70 (ddd, J=11.2, 7.8, 4.4 Hz, 1H), 2.69-2.58 (m, 1H), 2.56-2.46 (m, 1H), 2.38 (m, 2H), 2.11-2.01 (m, 3H), 2.00-1.82 (m, 7H), 1.79-1.65 (m, 4H), 1.65-1.56 (m, 2H), 1.53-1.45 (m, 5H), 1.42-1.21 (m, 10H), 1.04 (d, J=6.7 Hz, 3H), 0.89-0.98 (m, 12H), 0.85 (t, J=6.4 Hz, 6H), 0.64 (s, 3H).

(32) .sup.13C NMR (100 MHz, CDCl.sub.3): 140.99, 139.97, 135.63, 131.93, 119.47, 116.47, 104.05, 99.60, 88.10, 81.24, 75.07, 55.69, 54.50, 52.63, 46.05, 44.52, 42.81, 40.50, 39.07, 38.81, 37.97, 37.48, 37.26, 36.49, 34.72, 33.10, 30.77, 29.72, 28.35, 27.98, 26.25, 24.52, 24.70, 23.03, 21.13, 21.05, 20.40, 19.98, 19.67, 17.62, 16.21, 13.11, 12.07.

Example 7

(33) Synthesis of JZ-07

(34) ##STR00022##

(35) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and lithocholic acid (753 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound JZ-07 (843 mg, yield 65.5%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 12:1-2:1.

(36) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.47 (s, 1H), 4.92 (d, J=3.3 Hz, 1H), 3.73-3.53 (m, 1H), 2.66-2.56 (m, 1H), 2.39 (m, 2H), 2.27 (ddd, J=15.8, 9.7, 6.4 Hz, 1H), 2.05 (dt, J=14.5, 3.7 Hz, 1H), 1.99-1.94 (m, 1H), 1.93-1.70 (m, 8H), 1.68-1.55 (m, 3H), 1.51-1.21 (m, 21H), 1.08 (m, 4H), 1.00-0.89 (m, 13H), 0.66 (s, 3H).

(37) .sup.13C NMR (100 MHz, CDCl.sub.3): 180.33, 104.04, 100.16, 88.07, 81.26, 76.26, 56.42, 55.98, 52.63, 44.57, 42.77, 41.97, 40.42, 40.12, 37.48, 36.50, 35.85, 35.32, 35.42, 34.74, 32.59, 31.00, 30.85, 30.78, 29.72, 29.01, 28.20, 27.33, 26.40, 26.27, 24.70, 24.57, 24.21, 23.51, 20.89, 20.42, 18.30, 13.18, 12.07.

Example 8

(38) Synthesis of JZ-08

(39) ##STR00023##

(40) To a solution of dihydroartemisinin (569 mg, 2.0 mmol) and deoxycholic acid (785 mg, 2.0 mmol) in diethyl ether (50 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (1 mL) at 78 C. The temperature was naturally raised to ambient temperate and the reaction mixture was stirred overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (30 mL) was slowly added to quench the reaction, and then the reaction mixture was extracted with ethyl acetate (30 mL3). The organic phases were combined, washed with water (50 mL) and saturated NaCl solution (50 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound (723 mg, 54.8%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 12:1-3:1.

(41) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.47 (s, 1H), 4.91 (d, J=3.4 Hz, 1H), 4.02 (s, 1H), 3.67-3.55 (m, 1H), 2.65-2.56 (m, 1H), 2.48-2.25 (m, 3H), 2.07-2.01 (m, 1H), 1.92-1.81 (m, 5H), 1.77-1.70 (m, 3H), 1.68-1.58 (m, 4H), 1.54 (m, 2H), 1.50-1.34 (m, 12H), 1.27 (m, 8H), 1.18-1.06 (m, 2H), 1.01 (d, J=6.2 Hz, 3H), 0.96 (d, J=6.3 Hz, 3H), 0.93-0.85 (m, 7H), 0.71 (s, 3H).

(42) .sup.13C NMR (100 MHz, CDCl.sub.3): 179.27, 104.03, 100.49, 88.10, 81.30, 73.32, 52.62, 48.36, 47.33, 46.46, 44.55, 42.00, 37.43, 36.51, 36.06, 35.26, 35.04, 34.66, 34.30, 33.79, 32.64, 30.91, 30.82, 30.68, 29.72, 28.98, 27.43, 27.27, 26.29, 26.06, 24.69, 24.58, 23.62, 23.34, 20.38, 17.33, 13.20, 12.77.

Example 9

Synthesis of 10-(2-bromoethoxy)dihydroartemisinin

(43) To a solution of dihydroartemisinin (284 mg, 1.0 mmol) and 2-bromoethanol (125 mg, 1.0 mmol) in diethyl ether (20 mL) was slowly dropwise added BF.sub.3.Et.sub.2O (50 L) at 0 C. After the addition, the reaction mixture was stirred at 0 C. overnight. When the reaction was determined by TLC to be completed, a saturated aqueous NaHCO.sub.3 solution (10 mL) was slowly added to quench the reaction, and then the reaction mixture was extracted with ethyl acetate (10 mL3). The organic phases were combined, washed with water (20 mL) and saturated NaCl solution (20 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a white solid compound (203 mg, yield 51.9%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 30:1-8:1. .sup.1H NMR (400 MHz, CDCl.sub.3): 5.48 (s, 1H), 4.84 (d, J=3.3 Hz, 1H), 4.18-4.07 (m, 1H), 3.81-3.74 (m, 1H), 3.51 (t, J=5.2 Hz, 2H), 2.69-2.59 (m, 1H), 2.40-2.33 (m, 1H), 2.06-1.99 (m, 1H), 1.92-1.84 (m, 2H), 1.76 (m, 1H), 1.68-1.61 (m, 1H), 1.55-1.43 (m, 5H), 1.37-1.21 (m, 3H), 0.95-0.90 (m, 6H).

(44) .sup.13C NMR (101 MHz, CDCl.sub.3): 104.10, 102.02, 88.13, 81.08, 77.42, 77.11, 76.79, 68.16, 52.56, 44.35, 37.38, 36.39, 34.66, 31.47, 30.89, 26.16, 24.65, 24.36, 20.39, 13.00.

Example 10

(45) Synthesis of JZ-09

(46) ##STR00024##

(47) K.sub.2CO.sub.3 (276 mg, 2.0 mmol) and a catalytic amount of KI were added successively to a solution of 10-(2-bromoethoxy)dihydroartemisinin (391 mg, 1.0 mmol) and ursodeoxycholic acid (392 mg, 1.0 mmol) in DMF (20 mL) at room temperature. The reaction mixture was heated to 70 C. and reacted for 2 h. When the reaction was determined by TLC to be completed, water (20 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with ethyl acetate (20 mL3). The organic phases were combined, washed with water (30 mL) and saturated NaCl solution (50 mL) in sequence, dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound (536 mg, yield 76.2%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 4:1-1:1.

(48) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.45 (s, 1H), 4.84 (d, J=3.4 Hz, 1H), 4.28-4.21 (m, 2H), 4.08-3.98 (m, 1H), 3.68-3.57 (m, 3H), 2.72-2.58 (m, 1H), 2.43-2.34 (m, 2H), 2.28-2.19 (m, 1H), 2.04 (m, 2H), 1.91 (m, 2H), 1.85-1.77 (m, 5H), 1.71-1.60 (m, 5H), 1.53-1.44 (m, 11H), 1.39-1.25 (m, 12H), 1.19-1.03 (m, 3H), 0.94 (m, 12H), 0.69 (s, 3H).

(49) .sup.13C NMR (100 MHz, CDCl.sub.3): 174.08, 104.13, 102.08, 87.91, 81.09, 71.47, 71.40, 66.00, 63.38, 55.72, 54.85, 52.55, 44.40, 43.78, 42.43, 40.14, 39.17, 37.58, 37.30, 36.85, 36.43, 35.28, 34.93, 34.67, 34.09, 31.20, 30.99, 30.83, 30.35, 29.72, 28.63, 26.91, 26.21, 24.75, 24.41, 23.40, 21.18, 20.43, 18.40, 12.96, 12.15.

Example 11

(50) Synthesis of JZ-10

(51) ##STR00025##

(52) K.sub.2CO.sub.3 (276 mg, 2.0 mmol) and a catalytic amount of KI were added successively to a solution of 10-(2-bromoethoxy)dihydroartemisinin (391 mg, 1.0 mmol) and hyodeoxycholic acid (392 mg, 1.0 mmol) in DMF (20 mL) at room temperature. The reaction mixture was heated to 70 C. and reacted for 2 h. When the reaction was determined by TLC to be completed, water (20 mL) was slowly added to quench the reaction, and the reaction mixture was extractied with ethyl acetate (20 mL3). The organic phases were combined, washed with water (30 mL) and saturated NaCl solution (50 mL) in sequence, dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a colorless oily compound (589 mg, yield 83.7%), where an eluent was a mixture of petroleum ether and ethyl acetate in a ratio of 4:1-1:1.

(53) .sup.1H NMR (400 MHz, CDCl.sub.3): 5.45 (s, 1H), 4.84 (d, J=3.4 Hz, 1H), 4.28-4.22 (m, 2H), 4.10-4.00 (m, 2H), 3.67-3.62 m, 2H), 2.71-2.58 (m, 1H), 2.43-2.35 (m, 2H), 2.22 (s, 1H), 2.07 (m, 1H), 1.98-1.87 (m, 4H), 1.82-1.74 (m, 4H), 1.67-1.59 (m, 8H), 1.46-1.28 (m, 15H), 1.17-1.08 (m, 6H), 0.98-0.89 (m, 12H), 0.65 (s, 3H).

(54) .sup.13C NMR (100 MHz, CDCl.sub.3): 174.10, 104.12, 102.06, 87.90, 81.09, 71.60, 68.07, 66.00, 63.38, 56.14, 55.86, 52.55, 48.39, 44.40, 42.85, 39.95, 39.80, 37.58, 36.43, 35.96, 35.35, 35.55, 35.03, 34.83, 34.66, 31.19, 30.92, 30.83, 30.24, 29.20, 28.13, 26.20, 24.74, 24.41, 24.21, 23.50, 20.75, 20.42, 18.27, 12.95, 12.05.

Example 12

(55) Synthesis of JZ-11

(56) ##STR00026##

(57) To a solution of compound JZ-02 (130 mg, 0.18 mmol) in pyridine (2 mL) was added pyridine-sulfur trioxide complex (180 mg, 1.13 mmol) at 0 C. Then the reaction mixture was reacted at room temperature for 6 h until the reaction was determined by TLC to be completed. 1.0 M diluted hydrochloric acid (5 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with dichloromethane (10 mL3). The organic phases were combined, washed with 1.0 M diluted hydrochloric acid (20 mL2) and saturated NaCl solution (30 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a white solid compound JZ-11 (103 mg, 70.6%), where an eluent was a mixture of dichloromethane and methanol in a ratio of 20:1-10:1.

(58) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.97 (s, 1H), 5.34 (s, 1H), 4.80 (d, J=3.3 Hz, 1H), 3.94 (s, 1H), 3.39 (s, 1H), 2.42-2.30 (m, 1H), 2.29-2.04 (m, 5H), 2.01-1.89 (m, 2H), 1.84-1.54 (m, 10H), 1.38 (m, 8H), 1.27 (m, 7H), 1.21-1.10 (m, 6H), 1.00 (m, 2H), 0.92-0.81 (m, 12H), 0.61 (s, 3H).

(59) .sup.13C NMR (100 MHz, DMSO-d.sub.6): 175.89, 103.71, 100.13, 87.51, 80.98, 76.33, 55.29, 55.08, 44.40, 43.66, 42.24, 41.43, 37.25, 36.55, 35.28, 35.01, 34.60, 34.25, 34.15, 33.76, 31.44, 31.27, 30.94, 30.28, 29.49, 28.97, 26.13, 25.87, 24.73, 24.60, 23.74, 21.47, 20.65, 18.78, 13.40, 12.42.

Example 13

(60) Synthesis of JZ-12

(61) ##STR00027##

(62) To a solution of compound JZ-08 (130 mg, 0.18 mmol) in pyridine (2 mL) was added pyridine-sulfur trioxide complex (180 mg, 1.13 mmol) at 0 C. Then the reaction mixture was reacted at room temperature for 6 h until the reaction was determined by TLC to be completed. 1.0 M diluted hydrochloric acid (5 mL) was slowly added to quench the reaction, and the reaction mixture was extracted with dichloromethane (10 mL3). The organic phases were combined, washed with 1.0 M diluted hydrochloric acid (20 mL2) and saturated NaCl solution (30 mL), dried with anhydrous MgSO.sub.4 and filtered. The filtrate was rotatorily evaporated under vacuum to give a crude product, which was purified by silica gel column chromatography to give a white solid compound JZ-12 (112 mg, 76.8%), where an eluent was a mixture of dichloromethane and methanol in a ratio of 20:1-12:1.

(63) .sup.1H NMR (400 MHz, DMSO-d.sub.6): 11.75 (s, 1H), 5.33 (s, 1H), 4.79 (d, J=3.3 Hz, 1H), 4.34 (s, 1H), 3.46 (d, J=10.7 Hz, 1H), 2.37 (m, 1H), 2.27-2.11 (m, 3H), 2.08-1.92 (m, 2H), 1.83-1.57 (m, 10H), 1.53-1.43 (m, 5H), 1.40-0.97 (m, 18H), 0.96-0.78 (m, 14H), 0.64 (s, 3H).

(64) .sup.13C NMR (100 MHz, DMSO-d.sub.6): 177.08, 103.70, 98.99, 87.45, 81.01, 78.29, 76.61, 52.56, 48.62, 46.45, 46.07, 44.41, 42.18, 37.21, 36.54, 36.03, 35.70, 34.68, 34.50, 33.89, 32.51, 31.61, 30.89, 28.68, 27.70, 26.46, 26.12, 25.12, 24.73, 24.02, 23.79, 20.63, 17.77, 13.41, 12.73.

Example 14

(65) Inhibitory Effect of the Dihydroartemisinin-Steroid Conjugate on Various Tumor Cells Cultured In Vitro

(66) 1. Experimental Principle

(67) Effect of the compounds prepared in the above examples on the growth of tumor cells was investigated, where the tumor cells used herein were respectively human prostate cancer cell-PC-3 cells (CRL-1435), human cervical cancer cell line-HeLa cells (CCL-2), human acute T cell leukemia cells-Jurkat cells (TIB-152), human renal clear cell adenocarcinoma-786-0 cells (CRL-1932) and human breast cancer cells-MCF7 cells (HTB-22). African green monkey kidney cells-Vero cells (CCL-81) were used as normal cells to evaluate the cytotoxic effects of the compounds. Under the intervention of such compounds, the living cells can be stained using a cell proliferation-cytotoxicity assay kit Cell Counting Kit-8 (CCK-8 solution), and the OD value was measured to reflect the survival and growth state of the cells, that is, the cell survival rate (%) and the cell growth inhibition rate (%). Finally, a compound concentration at which only half of the cells survived, i.e., CC.sub.50, and a compound concentration at which a cell growth inhibition rate was 50%, i.e., IC.sub.50, were respectively calculated.

(68) 2. Experimental Materials

(69) Vero cells (CCL-81), PC-3 cells (CRL-1435), HeLa cells (CCL-2), Jurkat cells (TIB-152), 786-O cells (CRL-1932) and MCF7 cells (HTB-22) were all purchased from ATCC; DMEM, F12 medium, MEM and RPMI 1640 medium were purchased from Gibco (Life Technologies); fetal bovine serum (FBS) was purchased from Hyclone (Thermo Scientific); trypsin was purchased from Gibco (Life Technologies); CCK-8 solution was purchased from Shanghai Dojindo Chemical Technology Co., Ltd.; 96-well cell culture plate (with a transparent and flat bottom) was purchased from Corning; and SpectraMax 190 microplate reader was purchased from Molecular Devices Corporation.

(70) 3. Experimental Method

(71) The culture solution containing 10% FBS was regarded as a complete medium. Vero cells were cultured in DMEM complete medium; PC-3 cells were cultured in F12 complete medium; Hela cells were cultured in MEM complete medium; Jurkat cells were cultured in RPMI 1640 complete medium; 786-O cells were cultured in RPMI 1640 complete medium; and MCF-7 cells were cultured in MEM complete medium. After all the cells were cultured to logarithmic growth phase, the tests were performed as follows.

(72) The adherent cells were digested by pancreatin and suspended to directly collect cells. The cell suspension was centrifuged, and after counted, the cells were seeded in a transparent 96-well culture plate and adjusted to 510.sup.3 cells/100 L/well with respective complete mediums. At the same time, each plate was provided with blank wells without cells and samples and control wells with cells but without samples. The 96-well culture plates were then cultured overnight in a 37 C., 5% CO.sub.2 incubator. The test compounds were added at 50 L/well on the next day, and respective complete mediums were added at the same amount. The final concentrations of the compound samples were respectively 100 M, 25 M, 6.25 M, 1.56 M, 0.39 M, 0.1 M, 0.02 M and 0.006 M, and each concentration was repeated twice. While for the control wells and blank wells, complete mediums rather than the compound samples were added, where the complete culture medium was also added at 100 L/well. The culture plates were incubated in a 37 C., 5% CO.sub.2 incubator for 48 h. 2 h before the end of the incubation, the culture medium was removed, and complete culture medium containing 10% CCK-8 solution was added at 100 L/well. After the incubation, the plates were measured at 450 nM using a microplate reader for OD values (650 nM was used as reference).

(73) The CC.sub.50 was calculated according to the survival rate (%) of Vero cells to reflect the cytotoxic effect of the compounds. The IC.sub.50 was calculated according to the growth inhibition (%) of the tumor cells to reflect the antitumor activity of the samples.

(74) The survival rate and the growth inhibition were calculated as follows:
cell survival rate (%)=(OD value of the sample wellOD value of the blank well)/(OD value of the control wellOD value of the blank well)100%; and
cell growth inhibition rate (%)=[1(OD value of the sample wellOD value of the blank well)/(OD value of the control wellOD value of the blank well)]100%

(75) 4. Experimental Results

(76) The results of IC.sub.50 were shown in Tables 1 and 2. It can be seen from the tables that the dihydroartemisinin-steroid conjugate can inhibit the growth of various tumor cells. The potency of these compounds is far stronger than that of the dihydroartemisinin.

(77) TABLE-US-00001 TABLE 1 Inhibitory effect of dihydroartemisinin-steroid conjugate on various tumor cells Tumor cells Vero PC3 Hela Jurkat 786-O MCF7 A549 CC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 Compounds (M) (M) (M) (M) (M) (M) (M) JZ-1 13.58 0.63 0.39 2.15 0.20 4.13 0.1 JZ-2 7.81 0.26 0.04 0.24 0.06 3.39 0.02 JZ-3 6.55 0.53 0.22 0.12 0.034 1.08 0.01 JZ-4 100 25 25 32 100 35 2.40 JZ-5 100 100 100 45 100 56 3.20 JZ-6 100 100 100 34 100 45 6.33 JZ-7 100 100 100 54 100 65 3.70 JZ-8 6.5 0.01 0.45 0.02 0.006 0.01 0.006 JZ-9 20 1.05 1.36 0.05 0.11 0.23 0.28 JZ-10 7.1 0.8 1.01 0.34 0.09 2.05 0.05 Dihydroartemisinin 5.88 2.02 1.66 100 1.10 100 2.96 Taxol 1.5 0.006 0.006 0.03 0.006 0.01 0.006

(78) TABLE-US-00002 TABLE 2 Inhibitory effects of JZ-2 and JZ-8 on various tumor cells JZ-2 JZ-8 Taxol IC.sub.50 IC.sub.50 IC.sub.50 Cells (M) (M) (M) U87 0.517 0.200 0.010 SW1990 0.517 0.260 0.010 Kasumi 0.157 0.157 0.010 U2OS 0.357 0.420 0.010 OV-90 0.157 0.157 0.010 HC7116 0.157 0.167 0.010 DU-145 0.625 1.300 0.010 HEPG-2 0.500 0.500 0.010 RCC-4 2.500 0.625 10.000 RCC4 vhl 30.000 2.500 30.000 K562 0.157 0.157 0.010 NB4 0.157 0.157 0.010

Example 15

(79) Anti-Cancer Effect of JZ-2 on the Xenograft Model of Human Lung Cancer Cell A549

(80) 23 BALB/c nude mice, weighing 18-22 g and aged 2-4 weeks, were used in the experiment. Each nude mouse was subcutaneously injected with human non-small cell lung cancer cells A549 (1-510.sup.6), and when the tumor grew to 1-2 cm.sup.3 (about 40 days), the 23 nude mice were randomly divided into three groups, where group 1 (9 mice) was the control group, group 2 (9 mice) was the treatment group and group 3 (5 mice) was the positive control group (paclitaxel). Mice in group 1 were each intraperitoneally injected with 20 L of normal saline per day; mice in group 2 were each intraperitoneally injected with 10 mg/kg JZ-02 every day with a 2-day break after 7 consecutive days of administration; and mice in group 3 were each intraperitoneally injected with 10 mg/kg paclitaxel once every three days. The entire treatment lasted for 28 days, where three deaths occurred in group land one death occurred in group 3, while no death was observed in group 2, indicating that JZ-02 was less toxic than paclitaxel.

(81) It can be seen from the tumor growth curve shown in FIG. 1 that the compound JZ-2 of the invention can significantly inhibit the tumor growth, and the inhibitory effect was better than that of the positive control paclitaxel.

(82) It can be seen from the body weight curve shown in FIG. 2 that there was no significant decrease in body weight of the mice after 28 consecutive days of administration of JZ-2, demonstrating that the compound was less toxic and had no significant effect on the body weight and life quality of the mice.

(83) After the mice in each group were sacrificed, the tumor tissues were weighed for the calculation of the tumor inhibition rate, where the tumor inhibition rate of group 2 (the treatment group) was 58% and the tumor inhibition rate of the positive control group was 52%.

(84) Described are merely preferred embodiments, which are not intended to limit the invention. It should be noted that various variations and modifications made by those skilled in the art without departing from the spirit of the invention should fall within the scope of the invention defined by the appended claims.