CN114656435A - Rockmilan alcohol hydroxyl derivative, preparation method and application thereof - Google Patents

Abstract

The present invention discloses a roquemilanoyl hydroxyl derivative, a preparation method and application thereof, and has the following structural formula (I), wherein: R is a benzoic acid group, a p-fluorobenzoic acid group, a p-methoxybenzoic acid group, and a p-methoxybenzoic acid group. Bromobenzoic acid, 4,5-dimethoxy-2-nitro-benzoic acid, p-nitrocinnamate, 2-furanoic acid, 2-picolinic acid, 6-chloronicotinic acid, butyl Acid group, caprylic acid group, dimethylamino group, diethylamino group, dibutylamino group, dioctylamino group, benzylamino group, 3,4-dimethoxyphenethylamino group, 1-tetrahydropyrrole group, 1-imidazolyl, 1-pyrazolyl, 1-morpholinyl, 1-piperidinyl or 4-methylpiperazin-1-yl. The invention has nanomolar anti-colorectal cancer, anti-leukemia and anti-breast cancer activities.

Description

A kind of roquemilano alcohol hydroxyl derivative, its preparation method and application

technical field

The invention belongs to the technical field of medicines, and specifically relates to a roquemilanol hydroxy derivative, also relates to a preparation method of the roquemilanol hydroxy derivative, and the application of the roquemilanol hydroxy derivative in the preparation of antitumor drugs.

Background technique

Cancer or tumor is a common and frequently-occurring disease that seriously threatens human health, and currently the mortality rate has risen to the first place according to the statistics of the World Health Organization. There are three main methods of treatment for cancer or tumor: surgery, radiation and chemotherapy. However, the current clinical treatment is still largely dominated by chemotherapy. Existing chemotherapeutic drugs are one of the common methods for non-surgical treatment of cancer or tumors, and there are problems such as the close effective dose and toxic dose, and toxic and side effects. Or tumor patients are difficult to accept chemotherapy due to severe reaction or can not adhere to complete the entire course of treatment. In addition, radiotherapy and chemotherapy are not selective, have severe side effects on normal tissues, and induce drug resistance in cancer cells. The research on high-efficiency and low-toxicity anticancer drugs is the hotspot and focus of research at home and abroad.

The Wnt signaling pathway is a complex network of protein interactions whose functions are most commonly found in embryonic development and cancer, but are also involved in normal physiological processes in adult animals. In a variety of malignant tumors, the Wnt signaling pathway is often highly activated, especially in colorectal cancer, breast cancer and liver cancer. Recent studies have shown that the canonical Wnt/β-catenin signaling pathway plays a crucial role in regulating the self-renewal ability of tumor cells and maintaining the stemness of tumor cells. The discovery of anti-colorectal cancer drugs targeting the Wnt signaling pathway has become a research hotspot.

Chinese Invention Patent Publication No. CN113149942A disclosed on July 23, 2021 a kind of Rockmilanol phenolic hydroxyl derivative, its preparation method its structure is Rockmilanol phenolic hydroxyl derivative, and the main mechanism of action is MAPK signaling pathway inhibitor.

SUMMARY OF THE INVENTION

The purpose of the present invention is to overcome the above-mentioned shortcomings and provide a hydroxy derivative of roquemilanol with nanomolar anti-colorectal cancer, anti-leukemia and breast cancer activities.

Another object of the present invention is to provide a preparation method of the roquemiranol hydroxy derivative.

A further object of the present invention is to provide the application of the roquemilanol hydroxy derivative in the preparation of anti-colorectal cancer, anti-leukemia, MAPK signaling pathway and Wnt signaling pathway inhibitor drugs.

The purpose of this invention is to realize by following technical method:

A kind of roquemilano alcohol hydroxyl derivative of the present invention has the following structural formula (I):

Where: R is methyl, ethyl, propyl, butyl, pentyl, hexyl, 5-bromopentyl, allyl, isopentenyl, propargyl, 2-butynyl, acetyl, propyl Acyl, pentanoyl, benzoyl, p-nitrobenzoyl, ethylsulfonyl, propylsulfonyl or phenylsulfonyl.

The preparation method of a kind of loxanol hydroxy derivative of the present invention comprises the following steps:

(1) Synthesis of compound 3

Weigh 20g apigenin (compound 1, 0.0740mol) into a 500ml round-bottomed flask, add 51g anhydrous potassium carbonate (5.0eq) and 250ml acetone as a solvent to the flask, and slowly add 24.5 ml dimethyl sulfate under stirring (3.5eq), put the system into an oil bath and heated to reflux for 72h at 70°C. The reaction was followed by TLC. After the reaction was completed, it was cooled to room temperature and adjusted to pH=11 with ammonia water. The precipitate was removed by filtration, the filtrate was washed with a saturated saline solution, dried with anhydrous sodium sulfate, part of the solvent was spin-dried, silica gel was added to mix the sample, and purified using a flash silica gel column (chloroform:acetone=8:2) to obtain a pale yellow solid compound;

Take by weighing the compound 2 of 700mg, add 80mL of dichloromethane and 60mL of acetone mixed solvent, slowly add 140ml concentration of 11.6g/ml potassium hydrogen monosulfate complex salt aqueous solution, obtain compound 3, reaction scheme is as follows:

(2) Synthesis of a mixture of compounds 4 and 5

Weigh 800 mg of compound 3 (2.44 mmol), add 40 ml of acetonitrile, 30 ml of methanol and 5.0 g of 12.6 equivalents of methyl trans-cinnamate, irradiate with a xenon lamp, and react for 17 h to obtain a mixture of product compounds 4 and 5 , the reaction route is as follows:

(3) Synthesis of compound 6

Weigh 1 g of the mixture of compounds 4 and 5, add 30 mL of methanol and 10 mL of a methanol solution of 0.5 M sodium methoxide, and reflux at 70 °C for 4 h to obtain compound 6. The reaction scheme is as follows:

(4) Synthesis of compound 7

Weigh 2 g of compound 6, add 100 mL of DMSO and 170 mg of lithium chloride, and stir at 100°C for 8 h to obtain compound 7. The reaction scheme is as follows:

(5) Synthesis of compound 8

Weigh 3.27 g of sodium triacetoxyborohydride, add 200 mL of acetonitrile and 1.2 mL of glacial acetic acid, then slowly add 700 mg of compound 7, and stir at 40°C for 8 h to obtain compound 8. The reaction scheme is as follows:

(6) Synthesis of compound 9

Weigh 30mg of compound 8 (0.069mmol) into a 25ml round bottom flask, add 2ml DCM as solvent, add 28.7μl Et ₃ N (0.207mmol), 11μl chloroacetyl chloride (0.138mmol) and catalytic amount of DMAP, stir at room temperature 10h, to obtain compound 9, the reaction formula is as follows:

(7) Synthesis of compounds 10a to 10w

Weigh out 30 mg of 1-chloroacetyl-4'-demethoxylockmilanol (9, 0.059 mmol), add 5 ml of DMF solution, 16.3 mg of potassium carbonate (0.118 mmol), 1.5 equivalents of acid (0.085 mmol) ), heated in a constant temperature oil bath at 70°C for 12h to obtain a series of derivatives 10a to 10w, the reaction route is as follows:

The application of the hydroxy derivatives of locomilanol of the present invention in the preparation of anti-colorectal cancer, anti-leukemia and anti-breast cancer drugs.

The application of the roquemiranol hydroxy derivative of the present invention in the preparation of MAPK signaling pathway and Wnt signaling pathway inhibitor medicines.

Compared with the prior art, the present invention has obvious beneficial effects, as can be seen from the above technical solutions: the present invention takes apigenin as a raw material, and generates compound 2 under the action of dimethyl sulfate, which is It is oxidized to compound 3 under the action of photocatalytic [3+2] reaction between compound 3 and methyl cinnamate under the illumination of xenon light to obtain the tautomeric compound 4/5, and compound 4/5 undergoes a similar frequency in methanol-sodium methoxide solution. Alcohol rearrangement gives compound 6, compound 7 is demethoxylated under the action of lithium chloride to obtain compound 7, which is reduced by sodium triacetoxyborohydride to obtain compound 8, and compound 8 obtains active leader under the action of chloroacetyl chloride Under the action of different acids or amines, 1-Chloroacetyl Rockmilanol (Compound 9), 1-Chloroacetyl Rockmilanol (Compound 9) can obtain 1-Acid or amino-substituted Rockmilanol with different structures derivative. The compound of the present invention has nanomolar anti-colorectal cancer, anti-leukemia and anti-breast cancer activities, and induces colorectal cell apoptosis and cell cycle arrest by inhibiting MAPK and Wnt signaling pathways. When used as a medicine, it can be used directly or in the form of a pharmaceutical composition.

Description of drawings

Figure 1 is a graph showing the effect of compound 10r on the treatment of HCT116 tumor cells for 48 hours;

Figure 2 is a graph showing the percentages of compound 10r at different stages of the cycle of HCT116 tumor cells;

Figure 3 is a graph showing the apoptosis of HCT116 tumor cells induced by compound 10r;

Figure 4 is a bar graph of the apoptosis of tumor cells induced by compound 10r;

Figure 5 The effect of compound 10r on the formation of HCT116 clone spheres

Figure 6 Statistical graph of the number of HCT116 clone spheres formed by compound 10r

Figure 7 is a graph showing the effect of compound 10r on related apoptotic proteins;

Figure 8 is a graph showing the effect of compound 10r on related cyclins;

Figure 9 is a graph showing the effect of compound 10r on MAPK signaling pathway-related proteins;

Figure 10 is a graph showing the effect of compound 10r on Wnt signaling pathway-related proteins;

Detailed ways:

The substantive content of the present invention is further described below with the embodiments of the present invention, but the present invention is not limited by this.

Embodiment 1: the preparation method of target compound 10a, comprises the following steps:

(1) Synthesis of compound 3

Weigh 20g apigenin (compound 1, 0.0740mol) into a 500ml round-bottomed flask, add 51g anhydrous potassium carbonate (5.0eq) and 250ml acetone as a solvent to the flask, and slowly add 24.5 ml dimethyl sulfate under stirring (3.5eq), put the system into an oil bath and heated to reflux for 72h at 70°C. The reaction was followed by TLC. After the reaction was completed, it was cooled to room temperature and adjusted to pH=11 with ammonia water. The precipitate was removed by filtration, the filtrate was washed with a saturated saline solution, dried with anhydrous sodium sulfate, part of the solvent was spin-dried, silica gel was added to mix the sample, and purified using a flash silica gel column (chloroform:acetone=8:2) to obtain a pale yellow solid compound 2 ( 12g), 52.3% yield.

Weigh 700mg of compound 2 (2.24mmol), add it into a 1000mL round-bottomed flask, then add 80mL of dichloromethane and 60mL of acetone respectively, place it on a stirrer at room temperature for stirring and dissolving, then configure 200mL of buffer (16g sodium carbonate, 7.6 g of sodium bicarbonate, 200 mL of water), the prepared buffer solution was added to the round-bottomed flask, and the mixture was stirred for 30 minutes. Weigh 12 g of potassium hydrogen persulfate complex salt, add 140 ml of water to dissolve by ultrasonic. Place a dropping funnel on the round-bottomed flask, add 140 mL of the salt solution, and add slowly, about 5-7 seconds per drop. After the dropwise addition of the salt solution, the reaction solution was detected with pH test paper to make the reaction system pH=9. Stirring was carried out overnight and monitored by thin layer silica gel chromatography. Continue to configure 140 mL of potassium hydrogen peroxymonosulfate composite salt solution, and repeat the dropwise addition. After the dropwise addition of the salt solution, the reaction solution is detected with pH test paper, and adjusted to pH=9 with saturated aqueous sodium carbonate solution. Stirring was carried out again overnight and monitored by thin layer silica gel chromatography. Continue to repeat the previous experimental steps. Then turn off the normal temperature stirrer and let it stand for about 30 minutes. The dichloromethane layer was obtained by separation, and the organic layer was concentrated to obtain a small part of the concentrated solution, which was adjusted to pH=3 by adding p-toluenesulfonic acid monohydrate, and stirred at room temperature for 2 h. Thin layer silica gel chromatography monitoring followed by sample mixing. Purification by flash normal phase silica gel column with chloroform:methanol=50:1 gave compound 3 (350 mg) as a yellow solid in 50% yield.

(2) Synthesis of a mixture of compounds 4 and 5

Synthesis of a mixture of compounds 4 and 5: Weigh 800 mg of compound 3 (2.44 mmol) into a 250 mL round-bottom flask, add 40 mL of acetonitrile and 30 mL of methanol, and stir to dissolve. Then, 5.0 g of methyl trans-cinnamate was added, and the mixture was stirred and dissolved at room temperature. It was degassed with argon (Ar) using a diaphragm pump, with multiple air changes to ensure that the round-bottom flask was completely filled with argon. The round-bottomed flask was placed in a low temperature thermostatic stirrer at 0°C for stirring. The reaction was then irradiated with strong light using a xenon lamp. After stirring at 0°C for 17-20 h, the reaction was detected by thin-layer silica gel chromatography, and the reaction at the starting point was completed. The solvent was concentrated to dryness under the action of a water pump under reduced pressure using a rotary evaporator to obtain a mixture of compounds 4 and 5, which was directly used in the next step.

(3) Synthesis of compound 6

Weigh 1 g of the mixture of compounds 4 and 5 after the reaction in the previous step, add 30 mL of methanol to dissolve, stir at room temperature, slowly add 10 mL of methanol solution with a concentration of 0.5M sodium methoxide, and then place the system at 70 ° C. Oil bath reflux for 4h. The reaction was monitored by thin-layer silica gel chromatography. After the reaction was completed, the system was cooled to room temperature, 20 mL of saturated ammonium chloride solution was added to quench the reaction, and 100 mL of water was added. It was extracted three times with ethyl acetate, washed with saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate. It was then purified by flash column chromatography. It was eluted with a mixed solvent of chloroform:acetone 9:1 to obtain compound 6 as a brown oil, with a weight of 920 mg and a yield of 76.9%.

(4) Synthesis of compound 7

Weigh 2 g of compound 6 (0.0035 mol) into a 250 mL round-bottom flask, add 100 mL of DMSO to dissolve. Then add 170 mg of lithium chloride (1.1 equiv.), add 2 mL of water and stir at room temperature for 30 min, then place the round-bottomed flask in a constant temperature oil bath at 100 °C to heat overnight. The reaction was monitored by thin-layer silica gel chromatography until the reaction was completed. The reaction system was cooled to room temperature, 100 mL of water was added, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain an organic layer. Flash normal phase column chromatography was performed with a mixed solvent of petroleum ether:ethyl acetate=7:3 to obtain compound 7 (820 mg) as a yellow solid in a yield of 46.1%.

(5) Synthesis of compound 8

Weigh 3.27 g of sodium triacetoxyborohydride (9.0 equivalent) into a 250 mL round-bottom flask, add 200 mL of acetonitrile and 1.2 mL of glacial acetic acid, and stir at room temperature for 20 min. 700 mg of compound 7 (1.38 mmol) was dissolved in 90 mL of acetonitrile, and then slowly added dropwise to the reaction system. After stirring for 30 min, the round-bottomed flask was heated in a constant temperature oil bath at 40 °C. Monitor by thin-layer silica gel chromatography until the reaction is complete. 30 mL of saturated ammonium chloride solution was added to the reaction system to quench the reaction, then 100 mL of water was added, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the reactant. Normal phase column chromatography was performed with a mixed solvent of petroleum ether:ethyl acetate=1:0.5 to obtain compound 8 (420 mg) as a white solid in a yield of 69.2%.

Rockmillanol (8). White solid; melting point 136-137°C ¹ H NMR (400MHz, CDCl ₃ ) δ: 7.13-7.07 (5H, m), 6.99 (2H, d, J=7.1 Hz), 6.69-6.66 (2H,m), 6.29(1H,d,J＝1.9Hz), 6.14(1H,d,J＝1.9Hz), 4.81(1H,d,J＝6.3Hz), 4.00(1H,dd,J＝ 14.1, 6.5Hz), 3.90(3H,s), 3.84(3H,s), 3.71(3H,s), 3.30(1H,s), 2.74(1H,td,J=14.0,6.5Hz), 2.20( 1H, dd, J=14.0, 6.9 Hz), 1.72 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 163.9, 161.0, 158.6, 157.0, 138.7, 128.9, 128.9, 128.1, 128.1, 127.6, 127.6, 126.8, 126.2, 112.7, 107.7, 103.5, 94.8, 92.4, 89.4, 79.0, 55.8, 55.6, 55.0, 53.2, 36.4, 30.9; ESIMS m/z 457.0[M+Na] ⁺ ; HRESIMS m/z 457.1613[ M+Na] ⁺ (Calcd.for C ₂₆ H ₂₆ O ₆ Na, 457.1622).

(6) Synthesis of compound 9

Synthesis of compound 9: Weigh 30 mg of compound 8 (0.065 mmol) into a 25 ml round-bottomed flask, add 2 ml of DCM as a solvent, and stir at room temperature to dissolve the sample. 28.7 μl Et ₃ N (0.207 mmol), 11 μl chloroacetyl chloride (0.138 mmol) and catalytic amount of DMAP were added, stirred at room temperature for 10 h, and the reaction was followed by TLC dot plate until the starting point disappeared. Then 5 ml of water was added to quench the reaction, the DCM layer was separated, the aqueous layer was further extracted twice with 5 ml of DCM, the DCM layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, the solvent was spin-dried, and flash silica gel column chromatography ( Petroleum ether:ethyl acetate=6:4) to give white solid compound 9 (23.5 mg) in 69.5% yield.

1-Chloroacetyl-rockmilanol (9). White solid; melting point 173-174°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.15-7.12 (2H,m), 7.10-7.08 (5H,m) ,6.64(2H,d,J＝8.9Hz),6.23(1H,d,J＝1.9Hz),6.05(1H,d,J＝1.9Hz),5.94(1H,dd,J＝5.5,1.5Hz) ,4.06(1H,dd,J=13.7,6.2Hz),3.89(1H,d,J=15.1Hz),3.84(3H,s),3.78(3H,s),3.74(1H,d,J=15.1 Hz), 3.68 (3H,s), 2.88 (1H, td, J=13.8, 5.4Hz), 2.33 (1H, ddd, J=13.8, 6.2, 1.6Hz), 2.15 (1H,s); ¹³ C NMR (100MHz, CDCl ₃ )δ: 166.0, 163.8, 160.9, 158.6, 157.9, 138.2, 128.7, 128.7, 128.0, 128.0, 127.9, 127.9, 126.9, 126.4, 112.7, 106.4, 103.0, 9, 3.3, 91.8 55.6, 55.4, 55.0, 53.7, 40.9, 35.3, 30.9; ESIMS m/z 533.0[M+Na] ⁺ ; HRESIMS m/z 533.1330[M+Na] ⁺ (Calcd.for C ₂₈ H ₂₇ O ₇ ClNa, 533.1338 ).

(7) Synthesis of compound 10a

Weigh 30mg of compound 9 (0.0557mmol) into a 25ml round bottom flask, add 2ml (N,N-dimethylformamide) DMF as a solvent, then add benzoic acid 10.18mg (1.5eq), NaHCO ₃ 7mg ( 1.5eq), an appropriate amount of CsF, and then put the system into a 75°C oil bath to stir and react for 2 hours, TLC spot plate to detect the reaction, and stop the reaction until the raw material point disappears. Then 5 ml of water was added to quench the reaction, the DCM layer was separated, the aqueous layer was continuously extracted twice with 5 ml of ethyl acetate, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was spin-dried, and a flash silica column was used. Analysis (petroleum ether:ethyl acetate=6:4) gave 19.3 mg of white solid, which was the target compound 10a, and the yield was 48.7%.

1-(Benzoyl-2-hydroxyacetyl)-rockmilanol (10a) white solid; melting point 137-138°C; ¹ HNMR (400MHz, CDCl ₃ ) δ: 7.99 (2H, dd, J=8.6, 1.1 Hz), 7.58 (1H, t, J=7.4 Hz), 7.44 (2H, t, J=7.8 Hz), 7.15-7.06 (7H, m), 6.64 (2H, d, J=9.0 Hz), 6.14 ( 1H,d,J＝1.9Hz),6.05(1H,d,J＝1.9Hz),5.95(1H,dd,J＝5.3,1.2Hz),4.70(1H,d,J＝16.0Hz),4.61( 1H,d,J＝16.0Hz),4.02(1H,dd,J＝13.8,6.1Hz),3.82(3H,s),3.79(3H,s),3.68(3H,s),2.88(1H,td , J=13.8, 5.3 Hz), 2.36 (1H, ddd, J=13.8, 6.2, 1.4 Hz), 2.12 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.6, 165.7, 163.7, 160.7 ,158.6, 157.9,138.3,133.3,129.9,129.1,128.7,128.7,128.4,128.4,128.0,128.0,127.8,127.8,127.0,126.3,112.7,106.5,102.9,9.,5.5,91.8 , 55.5, 55.5, 55.0, 53.8, 35.5, 35.5; ESIMS m/z 619.2[M+Na] ⁺ ; HRESIMS m/z 631.1740[M+Cl] ^- (Calcd.for C ₃₅ H ₃₂ O ₉ Cl,631.1729) .

Example 2: Preparation of target compound 10b

The procedure was the same as that in Example 1, except that the benzoic acid was replaced with p-fluorobenzoic acid to prepare the target compound 10b in a yield of 58.9%.

1-(p-Fluorobenzoyl-2-hydroxyacetyl)-rocmilanol (10b). White solid; mp 159-160°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.03-7.96 (2H, m ),7.16–7.05(9H,m),6.64(2H,d,J=9.0Hz),6.13(1H,d,J=1.9Hz),6.04(1H,d,J=1.9Hz),5.94(1H ,d,J＝4.0 Hz),4.69(1H,d,J＝16.0Hz),4.60(1H,d,J＝16.0Hz),4.00(1H,dd,J＝13.9,6.1 Hz),3.82(3H ,s),3.81(3H,s),3.68(3H,s),2.88(1H,td,J=13.9,5.3Hz),2.35(1H,ddd,J=13.8,6.1,1.3Hz),2.10( 1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.5, 164.7, 163.7, 160.7, 158.6, 157.9, 138.2, 132.5, 132.4, 128.7, 128.7, 128.0, 128.0, 127.8, 127.8, 127.0, 1 125.4, 115.6, 115.5, 112.7, 106.5, 103.0, 93.2, 91.9, 88.2, 80.2, 60.9, 55.5, 55.5, 55.4, 55.0, 53.7, 35.5, 35.5; ESIMS m/z 637.3[M+Na] ⁺ ; HRESIMS m /z 649.1646[M+Cl] ^- (Calcd.forC ₃₅ H ₃₁ O ₉ ClF,649.1635).

Example 3: Preparation of target compound 10c

The steps were the same as those in Example 1, except that the benzoic acid was replaced with p-methoxybenzoic acid to prepare the target compound 10c with a yield of 55.9%.

1-(p-Methoxybenzoyl-2-hydroxyacetyl)-rocmilanol (10c). White solid; melting point 71-72°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.94 (2H, d , J＝8.6Hz), 7.14–7.06(7H,m), 6.90(2H,d,J＝8.6Hz), 6.64(2H,d,J＝8.6Hz), 6.15(1H,d,J＝1.9Hz) ), 6.05 (1H, d, J=1.9Hz), 5.94 (1H, dd, J=5.1, 1.4Hz), 4.67 (1H, d, J=16.0Hz), 4.57 (1H, d, J=16.0Hz) ),4.01(1H,dd,J＝13.7,6.1Hz),3.87(3H,s),3.82(3H,s),3.80(3H,s), 3.68(3H,s),2.87(1H,td, J=13.5, 5.3 Hz), 2.36 (1H, ddd, 13.8, 6.4, 1.7 Hz), 2.12 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.8, 165.4, 163.7, 163.6, 160.7, 158.6, 157.9,138.3,131.9,128.7,128.7,128.0,128.0,127.8,127.8,127.0,127.0,126.3,121.5,113.6,112.7,106.6,103.0,93.2,91.8,5.5.3,8 55.5, 55.4, 55.0, 53.7, 35.5, 35.5; ESIMS m/z 649.3[M+Na] ⁺ ; HRESIMS m/z 661.1845[M +Cl] ^- (Calcd. for C ₃₆ H ₃₄ O ₁₀ Cl,661.1835).

Example 4: Preparation of target compound 10d

The steps were the same as those in Example 1, except that the benzoic acid was replaced with p-bromobenzoic acid to prepare the target compound 10d in a yield of 58.5%.

1-(p-Bromobenzoyl-2-hydroxyacetyl)-rocmilanol (10d), white solid; mp 134-135°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.82 (2H, d, J =8.2Hz),7.52(2H,d,J=8.2Hz),7.15(2H,t,J=7.4Hz),7.10-7.05(5H,m),6.65(2H,d,J=8.7Hz), 6.11(1H,d,J＝1.9Hz),6.02(1H,d,J＝1.9Hz),5.93(1H,dd,J＝5.1,1.2Hz),4.69(1H,d,J＝16.0Hz), 4.60(1H,d,J＝16.0Hz),3.98(1H,dd,J＝13.9,6.1Hz),3.81(3H,s),3.79(3H,s),3.68(3H,s),2.88(1H , td, J=13.7, 5.3 Hz), 2.35 (1H, ddd, J=13.6, 6.2, 1.7 Hz), 2.11 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.3, 164.9, 163.7 , 160.7,158.6, 157.9,138.2,131.7,131.3,128.7,128.5,128.5, 128.0, 128.0, 128.0, 127.9,127.0, 126.7, 106.9.9,9.9,9,9,9,9,9.9,9.9. , 80.2, 61.0, 55.5, 55.5, 55.4, 55.0, 53.7, 35.5; ESIMS m/z 697.2[M+Na] ⁺ ; HRESIMS m/z 697.1050[M+Na] ⁺ (Calcd.for C ₃₅ H ₃₁ O ₉ BrNa, 697.1044).

Example 5: Preparation of target compound 10e

The steps were the same as those in Example 1, except that the benzoic acid was replaced with 4,5-dimethoxy-2-nitro-benzoic acid to prepare the target compound 10e with a yield of 58.8%.

1-(4,5-Dimethoxy-2-nitro-benzoic acid-2-hydroxyacetyl)-rocmilanol (10e). White solid; mp 145-146°C; ¹ H NMR (400MHz, CDCl) ₃ ) δ: 7.45(1H,s), 7.15–7.07(8H,m), 6.64(2H,d,J=8.8Hz), 6.16(1H,d,J=1.9Hz), 6.01(1H,d, J＝1.9Hz), 5.96(1H,d,J＝4.0Hz), 4.66(1H,d,J＝16.0Hz), 4.58(1H,d,J＝16.0Hz), 4.04(1H,dd,J＝ 13.6,6.2Hz),3.98(3H,s),3.97(3H,s),3.80(3H,s),3.78(3H,s),3.68(3H,s),2.89(1H,td,J=13.4 , 5.8Hz), 2.37 (1H, ddd, J=14.1, 6.1, 1.7Hz), 2.13 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.1, 164.7, 163.7, 160.6, 158.5, 157.9 ,152.4,150.6, 141.0,138.1,128.7,128.7,127.9,127.9,127.8,127.8,126.9,126.3,120.5,112.7, 111.0,116.9,106.4,102.9,93.2,9,6.6,58.3,5.6 , 56.5, 55.5, 55.4, 55.0, 53.7, 35.4; ESIMS m/z 724.2[M+Na] ⁺ ; HRESIMS m/z 724.2010[M+Na] ⁺ (Calcd.for C ₃₇ H ₃₅ O ₁₃ NNa,724.2000).

Example 6: Preparation of target compound 10f

The steps were the same as in Example 1, except that the benzoic acid was replaced with p-nitrocinnamic acid to prepare the target compound 10f with a yield of 54.8%.

1-(p-Nitrocinnamoyl-2-hydroxyacetyl)-rocmilanol (10f). White solid; melting point 110-111°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.26 (2H, d, J =8.8Hz), 7.73–7.65(3H,m), 7.15–7.06(7H,m), 6.65(2H,t,J=6.0Hz), 6.53(1H,d,J=16.1Hz), 6.17(1H ,d,J=1.9Hz),6.08(1H,d,J=1.9Hz),5.93(1H,d,J=4.0Hz),4.60(1H,d,J=16.0Hz), 4.42(1H,d , J=16.0Hz), 4.03(1H,dd,J=13.9,6.1Hz), 3.83(3H,s), 3.77(3H,s), 3.68(3H,s), 2.89(1H,td,J= 14.0, 5.3 Hz), 2.37 (1H, ddd, J=14.1, 6.1, 1.7 Hz), 2.11 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 166.3, 165.0, 163.7, 160.8, 158.6, 158.0, 148.6,142.9,140.2,138.2,128.8,128.8,128.7,128.7,127.9,127.9,127.9,126.9,126.4,124.1,121.0,112.0,106.6,103.0,9,7.2,91.8,7.6 60.8, 55.5, 55.4, 55.0, 55.0, 53.7, 35.5; ESIMS m/z 690.2[M+Na] ⁺ ; HRESIMS m/z 690.1954 [M+Na] ⁺ (Calcd.for C ₃₇ H ₃₃ O ₁₁ NNa, 690.1946).

Example 7: Preparation of target compound 10 g

The steps were the same as those in Example 1, except that benzoic acid was replaced with p-2-furancarboxylic acid to prepare 10 g of the target compound with a yield of 45.7%.

1-(2-furoyl-2-hydroxyacetyl)-rocmilanol (10 g). White solid; melting point 157-158°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.60 (1H, d, J =0.8Hz),7.20(1H,dd,J=3.6,0.8 Hz),7.15–7.05(7H,m),6.64(2H,d,J=9.0Hz),6.52(1H,dd,J=3.5, 1.7Hz), 6.17 (1H,d,J＝1.9Hz),6.07(1H,d,J＝1.9Hz),5.93(1H,dd,J＝5.3,1.Hz),4.64(1H,d,J =16.0Hz),4.55(1H,d,J=16.0Hz),4.03(1H,dd,J=13.8,6.1Hz),3.83(3H,s), 3.82(3H,s),3.68(3H,s ), 2.88 (1H, td, J=13.9, 8.6Hz), 2.36 (1H, ddd, J=13.8, 6.2, 1.4Hz), 2.12 (1H, s); ¹³ C NMR (100MHz, CDCl ₃ )δ: 166.2,163.8,160.7,158.6, 157.9,157.5,146.9,143.6,138.3,128.0, 128.0,127.0,127.0,126.3, 126.3,111.9.9, 106.5,9.8,9.2,9.2,9.2,9.2,9.2,9.2,9.2,9.2,9.2,9.9.2,9.8,9.8,9.8,9.8,9.8,9.8,9.8,9.9.9.9.8.8.8.8.8.8.8.8.8, 19.8.8.8.5,11.9.5. 80.2, 60.5, 55.5, 55.5, 55.0, 53.7, 35.5, 30.9; ESIMS m/z 609.2[M+Na] ⁺ ; HRESIMS m/z 587.1883[M+H] ⁺ (Calcd.for C ₃₃ H ₃₁ O ₁₀ , 587.1912).

Example 8: Preparation of target compound 10h

The steps were the same as those in Example 1, except that the benzoic acid was replaced with p-2-picolinic acid, and the target compound was prepared for 10 h with a yield of 59.8%.

1-(2-Pyridinyl-2-hydroxyacetyl)-rocmilanol (10h). White solid; mp 79-80°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.76 (1H, dd, J =4.7,0.7Hz),8.09(1H,d,J=7.8Hz),7.85(1H,t,J=6.92Hz),7.50(1H,ddd,J=8.16,4.87,1.36Hz),7.14–7.07 (7H,m), 6.64(2H,d,J＝6.1Hz), 6.14(1H,d,J＝1.9Hz), 6.05(1H,d,J＝1.9Hz), 5.94(1H,d,J＝ 4.0Hz), 4.71(1H,d,J＝15.9Hz),4.66(1H,d,J＝15.9Hz),4.05(1H,dd,J＝13.9,6.1Hz),3.83(3H,s),3.80 (3H,s),3.67(3H,s),2.92(1H,td,J=14.3,5.3),2.38(1H,ddd,J= ^{14.1,6.1,1.7Hz} ),2.12(1H,s); C NMR (100MHz, CDCl ₃ ) δ: 166.1, 164.1, 163.7, 160.7, 158.5, 157.9, 149.9, 147.0, 138.2, 136.9, 128.7, 127.9, 127.9, 127.8, 127.8, 127.2, 127.0, 126.3, 127.0, 126.3, 127.0 106.4, 102.9, 93.2, 91.7, 88.2, 80.2, 61.4, 55.5, 55.4, 55.00, 55.0, 53.7, 35.5; ESIMS m/z 620.2[M+Na] ⁺ ; HRESIMS m/z 620.1901[M+Na] ⁺ ( Calcd. for C ₃₄ H ₃₁ O ₉ NNa, 620.1891).

Example 9: Preparation of target compound 10i

The steps were the same as those in Example 1, except that the benzoic acid was replaced with 6-chloronicotinic acid to prepare the target compound 10i in a yield of 50.6%.

1-(6-Chloronicotinoyl-2-hydroxyacetyl)-lockmiranol (10i). White solid; melting point 79-80°C; ¹ HNMR (400 MHz, CDCl ₃ ) δ: 8.89 (1H, dd, J= 2.4,0.6Hz),8.15(1H,dd,J=8.3,2.4Hz),7.41(1H,dd,J=8.3,0.5Hz),7.17-7.05(7H,m),6.63(2H,d,J ＝9.0Hz), 6.10(1H,d,J＝1.9Hz),6.00(1H,d,J＝1.9Hz),5.91(1H,d,J＝4.2Hz),4.72(1H,d,J＝16.0 Hz), 4.63(1H,d,J＝16.0Hz),3.98(1H,dd,J＝14.0,6.1Hz),3.80(3H,s), 3.79(3H,s),3.67(3H,s), 2.89 (1H, td, J=14.0, 5.2 Hz), 2.37 (1H, dd, J=14.0, 6.1, 1.2 Hz), 2.08 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 165.9, 163.7,163.5,160.7, 158.6,157.8,156.0,151.2,139.6,138.1,128.6,127.9,127.8,126.8, 126.8,124.0,112.7,06.9,9.9,9,9.2,9.9,9,9.2,9.9,9.9,9,9.2,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9,9.9. 80.3, 61.1, 55.5, 55.5, 55.4, 55.0, 53.7, 35.5; ESIMS m/z 654.2[M+Na] ⁺ ; HRESIMS m/z 654.1510[M+ Na] ⁺ (Calcd.for C ₃₄ H ₃₀ O ₉ NClNa, 654.1501).

Example 10: Preparation of target compound 10j

The steps were the same as those in Example 1, except that the benzoic acid was replaced with butyric acid to prepare the target compound 10j with a yield of 52.3%.

1-(Butyryl-2-hydroxyacetyl)-rockmilanol (10j). White solid; mp 114-115°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.15-7.08 (7H, m), 6.64 (2H,d,J＝8.9Hz),6.23(1H,d,J＝1.9Hz),6.07(1H,d,J＝1.9Hz),5.91(1H,dd,J＝5.2,1.3Hz),4.45 (1H,d,J＝16.0Hz),4.35(1H,d,J＝16.0Hz),4.02(1H,dd,J＝13.7,6.2Hz),3.84(3H,s),3.79(3H,s) ,3.68(3H,s),2.86(1H,td,J＝14.7,5.4Hz),2.33(1H,ddd,J＝13.7,7.7,1.3 Hz),2.29(2H,td,J＝7.5,3.0Hz) ), 2.12 (1H, s), 1.64 (2H, dd, J=14.8, 7.4 Hz), 0.96 (3H, t, J=7.4 Hz); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 172.7, 166.7, 163.8, 160.8, 158.6, 158.0, 138.3, 128.7, 128.7, 128.0, 128.0, 127.8, 127.8, 127.0, 127.0, 126.3, 112.7, 106.6, 103.0, 93.2, 91.8, 55.2, 80.1, 6 53.7, 35.5, 35.5, 18.2, 13.6; ESIMS m/z 585.3[M+Na] ⁺ ; HRESIMS m/z 585.2104[M+Na] ⁺ (Calcd.for C ₃₂ H ₃₄ O ₉ Na,585.2095).

Example 11: Preparation of target compound 10k

The steps were the same as those in Example 1, except that the benzoic acid was replaced by capric acid to prepare the target compound 10k with a yield of 55.1%.

1-(Decanoyl-2-hydroxyacetyl)-rockmilanol (10k). White solid; mp 74-75°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.14-7.07 (7H, m), 6.65 -6.63(2H,m),6.22(1H,d,J=1.9Hz),6.07(1H,d,J=1.9Hz),5.91(1H,dd,J=5.2,1.4Hz),4.44(1H, d, J＝16.0 Hz), 4.35 (1H, d, J＝16.0 Hz), 4.02 (1H, dd, J＝13.6, 6.3 Hz), 3.84 (3H, s), 3.79 (3H, s), 3.68 ( 3H, s), 2.86 (1H, td, J=13.7, 5.4Hz), 2.34–2.28 (3H, m), 2.11 (1H, s), 1.66– 1.62 (4H, m), 1.28 (10H, d, J=14.4 Hz), 0.88 (3H, t, J=7.0 Hz); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 172.8, 166.7, 163.7, 160.7, 158.6, 158.0, 138.2, 128.7, 128.0, 128.0, 127.85,127.85,127.02,126.33,112.73,106.61,103.00,93.24,91.84,88.25,80.07,60.2,55.5,55.4,55.0,53.7,35.4,33.7,33.7,9.0.29.9,3 24.7, 22.6, 14.1; ESIMS m/z 669.3[M+Na] ⁺ ; HRESIMS m/z 647.3184[M+H] ⁺ (Calcd. for C ₃₈ .H ₄₇ .O ₉ ,647.3215).

Example 12: Preparation of target compound 10l

The steps were the same as those in Example 1, except that the benzoic acid was replaced with dimethylamine to prepare the target compound 10l with a yield of 59.4%.

1-(2-Methylaminoacetyl)-rockmilanol (10l). White solid; melting point 149-150°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.13-7.06 (7H, m), 6.63 ( 2H,d,J＝8.9Hz),6.23(1H,d,J＝1.9Hz),6.03(1H,d,J＝1.9Hz),5.86(1H,d,J＝3.7Hz),4.10(1H, dd,J＝13.8,6.1Hz), 3.84(3H,s), 3.76(3H,s), 3.67(3H,s), 3.00(1H,d,J＝16.9Hz), 2.86(1H,td,J = 13.9, 4.9 Hz), 2.82 (1H, d, J=16.9 Hz), 2.33 (1H, dd, J=13.7, 6.3 Hz), 2.25 (6H, s), 1.77 (1 H, s); ¹³ C NMR (100MHz, CDCl ₃ )δ169.3,163.7,161.0,158.5,157.9,138.5, 128.7,128.7,128.0,128.0,127.8,127.8,127.2,127.2,126.2,112.6,106.6,103.0,8.2,7.3 59.8, 55.6, 55.4, 55.0, 53.8, 44.9, 44.9, 35.7; ESIMS m/z 542.2[M+ Na] ⁺ ; HRESIMS m/z 520.2338[M+H] ⁺ (Calcd.for C ₃₀ H ₃₄ O ₇ N , 520.2329).

Example 13: Preparation of target compound 10m

The steps were the same as those in Example 1, except that the benzoic acid was replaced with diethylamine to prepare the target compound 10m in a yield of 56.6%.

1-(2-Diethylaminoacetyl)-rockmilanol (10m). White solid; melting point 50-51°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.10 (7H, m), 6.63 (2H, d,J＝9.0Hz),6.23(1H,d,J＝1.9Hz), 6.03(1H,d,J＝1.9Hz),5.82(1H,d,J＝3.6Hz),4.14(1H,dd, J=14.0, 6.2Hz), 3.83(3H,s), 3.76(3H,s), 3.67(3H,s), 3.15(1H,d,J=17.1Hz), 3.03(1H,d,J=17.1 Hz), 2.88 (1H, td, J=13.9, 4.9Hz), 2.54–2.48 (4H, m), 2.37 (1H, ddd, J=13.4, 6.1, 1.5Hz), 1.25 (1H, s), 0.96 (6H, t, J=7.2 Hz); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 170.2, 163.7, 161.1, 158.5, 158.0, 138.6, 128.6, 128.6, 128.0, 128.0, 127.8, 127.8, 127.3, 127.3, 126.2, 126.2, 112.6, 103.1, 93.3, 91.7, 88.2, 79.0, 55.6, 55.4, 55.0, 53.8, 53.8, 53.6, 47.4, 35.8, 12.4, 12.4; ESIMSm/z 548.3[M+H] ⁺ ; HRESIMS m/ z 548.2650[M+H] ⁺ (Calcd. for C ₃₂ H ₃₈ O ₇ N,548.2643).

Example 14: Preparation of target compound 10n

The steps were the same as those in Example 1, except that the benzoic acid was replaced with dibutylamine to prepare the target compound 10n with a yield of 48.2%.

1-(2-Dibutylaminoacetyl)-rockmilanol (10n). White solid; melting point 39-40°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.15-7.13 (3H, m), 7.10 -7.07(4H,m),6.64(2H,dd,J＝6.8,2.18Hz),6.22(1H,dd,J＝6.7,2.0Hz),6.05(1H,dd,J＝17.7,1.9Hz), 5.97 (1H, dd, J＝5.1, 1.6Hz), 4.45 (1H, d, J＝16.2Hz), 4.35 (1H, d, J＝16.2Hz), 4.09 (1H, dd, J＝13.8, 6.1Hz) ), 3.84(3H,s), 3.78(3H,s), 3.68(3H,s), 3.14(1H,d,J=16.8Hz), 2.91-2.82(1H,m), 2.42(1H,td, J=4.5, 2.3Hz), 2.37-2.33(1H,m), 2.17(1H,s), 1.53-1.46(2H,m), 1.35-1.21(7H,m), 0.96-0.85(7H,m) ; ¹³ C NMR (100MHz, CDCl ₃ )δ: 167.6, 163.7, 163.7, 161.1, 160.7, 158.6, 158.5, 158.0, 128.7, 128.6, 128.0, 127.8, 127.8, 126.2, 126.2, 112.7, 91.7, 3.1, 1 88.3, 88.1, 79.8, 61.1, 55.5, 55.5, 55.4, 55.3, 55.0, 55.0, 53.8, 53.8, 53.7, 29.9, 20.4, 14.0, 13.8; ESIMS m/z 604.2[M+ H] ⁺ ; HRESIMS m/z 604.3273 [M+H] ⁺ (Calcd.for C ₃₆ H ₄₆ O ₇ N,604.3269).

Example 15: Preparation of target compound 10o

The steps were the same as those in Example 1, except that the benzoic acid was replaced with dioctylamine to prepare the target compound 10o with a yield of 46.3%.

1-(2-Dioctylaminoacetyl)-rockmilanol (10o). White solid; melting point 41-42°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.14-7.11 (4H, m), 7.08 –7.05(3H,m),6.62(1H,d,J=8.9Hz),6.22(1H,dd,J=6.1,1.9Hz),6.02(1H,d,J=8.9Hz),5.82(1H, d,J＝3.8 Hz),4.13(1H,dd,J＝14.0,6.1Hz),3.83(3H,s),3.75(3H,s),3.67(3H,s),3.14(1H,d,J =17.0Hz),3.04(1H,d,J=17.0Hz),2.87(1H,td,J=13.1,6.5H,),2.40(4H,dd,J=8.1,5.4Hz),2.35(1H, ddd, J=13.5, 6.1, 1.0 Hz), 1.32 (5H, dd, J=14.5, 7.4 Hz), 1.28–1.22 (18H, m), 0.87 (7H, t, J=7.1 Hz); ¹³ C NMR (100MHz, CDCl ₃ )δ: 170.4, 163.7, 161.1, 158.5, 158.0, 138.6, 128.7, 128.0, 128.0, 127.8, 127.3, 126.2, 112.7, 106.6, 103.1, 93.4, 91.8, 88.5, 78.9, ESIMS m/z 738.5[M+Na] ⁺ ; HRESIMS m/z 716.4535[M+H] ⁺ (Calcd. for C ₄₄ H ₆₂ O ₇ N, 716.4521).

Example 16: Preparation of target compound 10p

The steps were the same as those in Example 1, except that the benzoic acid was replaced with benzylamine to prepare the target compound 10p with a yield of 55.6%.

1-(2-Benzylaminoacetyl)-rockmilanol (10p). White solid; melting point 59-60°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.31-7.27 (4H, m), 7.26 (1H,s), 7.15–7.06(8H,m), 6.67–6.62 (2H,m), 6.21(1H,d,J=1.9Hz), 5.99(1H,d,J=1.9Hz), 5.88( 1H,d,J＝3.6Hz), 4.08(1H,dd,J＝13.7,6.2Hz), 3.80(3H,s), 3.70(2H,s), 3.69(3H,s), 3.68(3H,s) ), 3.27(1H,d,J＝17.5Hz),3.12(1H,d,J＝17.5Hz),2.88(1H,td,J＝13.8,5.2Hz), 2.37–2.30(1H,m); ¹³ CNMR (100MHz, CDCl ₃ )δ: 170.9, 163.7, 160.9, 158.5, 157.9, 138.4, 128.7, 128.4, 128.3, 128.0, 128.0, 127.8, 127.8, 127.2, 127.1, 126.3, 112.7, 103.6.5,. , 88.3, 79.6, 55.5, 55.3, 55.3, 55.0, 53.8, 53.1, 49.8, 49.8, 35.5, 35.5, 29.3, 29.3; ESIMS m/z604.2[M+Na] ⁺ ; HRESIMS m/z 582.2484[M+ H ] ⁺ (Calcd.for C ₃₅ H ₃₆ O ₇ N,582.2486).

Example 17: Preparation of target compound 10q

The procedure was the same as that in Example 1, except that the benzoic acid was replaced with 3,4-dimethoxyphenethylamine to prepare the target compound 10q with a yield of 58.4%.

1-(2-(3,4-Dimethoxyphenethylamine)-acetyl)-rockmilanol (10q). White solid; mp 54-55°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.16-7.15(7H,m), 6.77(1H,d,J=8.2Hz), 6.73-6.71(2H,m), 6.65-6.62(2H,m), 6.22(1H,d,J=1.9Hz) ,6.00(1H,d,J＝1.9 Hz),5.85(1H,dd,J＝4.9,1.4Hz),4.08(1H,dd,J＝13.7,6.2Hz),3.86(3H,s),3.83 ( 3H,s), 3.82(3H,s), 3.70(3H,s), 3.68(3H,s), 3.25(1H,d,J=17.6Hz), 3.09(1H,d, J=17.6Hz), 2.86(1H,td,J=13.8,5.1Hz),2.80–2.71(2H,m),2.73-2.66(2H,m), 2.33(1H,ddd,J= ^{13.6,6.2,1.5Hz} ); NMR (100MHz, CDCl ₃ )δ: 171.1, 163.7, 160.9, 158.6, 157.9, 148.9, 147.4, 138.4, 132.2, 128.7, 128.7, 128.0, 128.0, 1127.8, 127.8, 127.1, 126, 3, 120.9, 112.3, 120.4, 112.3 , 106.6, 103.0, 93.3, 91.8, 88.2, 79.5, 55.9, 55.8, 55.6, 55.4, 55.4, 55.0, 53.8, 50.8, 50.8, 36.0, 35.6; ESIMS m/z 678.3[M+ Na] ⁺ ; HRESIMS m/z 656.2857[M+H] ⁺ (Calcd.for C ₃₈ H ₄₂ O ₉ N,656.2854).

Example 18: Preparation of target compound 10r

The procedure was the same as that of Example 1, except that the benzoic acid was replaced by tetrahydropyrrole, and the target compound 10r was prepared in a yield of 57.2%.

1-(2-(1-Tetrahydropyrrolyl)-acetyl)-rockmilanol (10r). Yellow solid; mp 121-122°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.16-7.05 (7H ,m),6.63(2H,d,J＝8.9Hz),6.23(1H,d,J＝1.9Hz),6.03(1H,d,J＝1.9Hz),5.86(1H,d,J＝3.7Hz) ), 4.12–4.05(1H,m), 3.83(3H,s), 3.76(3H,s), 3.67(3H,s), 3.19(1H,d,J=17.0Hz), 2.99(1H,d, J＝17.0Hz), 2.86(1H,td,J＝13.8,5.1Hz),2.59-2.47(4H,m),2.33(1H,ddd,J＝13.7,6.2,1.4Hz),1.80-1.75(3H , m); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 169.5, 163.6, 160.9, 158.5, 157.9, 138.5, 128.7, 128.7, 128.0, 128.0, 127.8, 127.8, 127.2, 126.2, 112.6, 106.7, 103 , 91.7, 88.1, 79.2, 56.2, 56.2, 55.5, 55.5, 55.4, 55.4, 54.9, 53.7, 53.6, 35.6, 35.6, 23.7, 23.7; ESIMS m/z546.3[M+H] ⁺ ; HRESIMS m/z 546.2485[M+H] ⁺ (Calcd.for C ₃₂ H ₃₆ O ₇ N,546.2486).

Example 19: Preparation of target compound 10s

The steps were the same as those in Example 1, except that the benzoic acid was replaced with imidazole to prepare the target compound 10s with a yield of 49.3%.

1-(2-(1-imidazolyl)-acetyl)-rockmilanol (10s). White solid; melting point 100-101°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.63 (1H, s), 7.14–7.01(8H,m),6.78(1H,s),6.62(2H,d,J=8.9Hz),6.27(1H,d,J=1.8Hz),6.08(1H,d,J=1.8Hz) ), 5.89(1H,d,J＝3.8 Hz),4.50(1H,d,J＝17.6Hz),4.28(1H,d,J＝17.6Hz),4.13-4.06(1H,m),3.87(3H , s), 3.71 (3H, s), 3.67 (3H, s), 2.90 (1H, td, J=13.6, 4.5Hz), 2.37 (1H, dd, J=13.7, 6.0Hz), 2.04 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 164.0, 161.2, 158.6, 157.9, 138.0, 133.3, 128.6, 128.6, 127.9, 127.9, 127.9, 126.8, 126.3, 112.7, 105.9, 102.9, 1.83.3 88.6, 80.8, 80.0, 65.1, 55.8, 55.6, 55.3, 55.0, 53.7, 53.4, 52.0, 48.2, 45.6; ESIMS m/z 533.2[M+H] ⁺ ; HRESIMS m/z 543.2124[M+H] ⁺ ( Calcd. for C ₃₁ H ₃₁ O ₇ N ₂ , 543.2125).

Example 20: Preparation of target compound 10t

The steps were the same as those in Example 1, except that the benzoic acid was replaced with pyrazole to prepare the target compound 10t with a yield of 58.7%.

1-(2-(1-Pyrazolyl)acetyl)-rockmiranol (10t). White solid; mp 95-96°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.48 (1H, d, J =1.6Hz),7.25(1H,d,J=2.2Hz),7.17–7.06(7H,m),6.63(2H,d,J=8.9Hz),6.26(1H,t,J=2.1Hz), 6.24(1H,d,J＝1.9Hz), 6.08(1H,d,J＝1.8Hz), 5.90(1H,d,J＝3.6Hz), 4.71(1H,d,J＝17.5Hz), 4.60( 1H, d, J=17.5Hz), 4.01 (1H, dd, J=13.7, 6.1Hz), 3.86 (3H, s), 3.79 (3H, s), 3.68 (3H, s), 2.86 (1H, td , J=13.8, 5.1Hz), 2.35 (1H, ddd, J=13.7, 6.2, 1.5Hz), 2.14 (1H, s); ¹³ C NMR (100MHz, CDCl ₃ )δ: 166.5, 163.9, 161.0, 158.6 ,158.0,139.9,138.3,130.4, 128.9,128.7,128.1,128.0,127.8,127.0,126.3,112.7,106.5,106.4,103.0,94.8,93.3,91.9,88.4,80.3.,55.6,55 , 52.9, 35.4; ESIMS m/z 565.2[M+Na] ⁺ ; HRESIMS m/z 565.1953[M+Na] ⁺ (Calcd.forC ₃₁ H ₃₀ O ₇ N ₂ Na, 565.1945).

Example 21: Preparation of target compound 10u

The steps were the same as those in Example 1, except that the benzoic acid was replaced with morpholine to prepare the target compound 10u with a yield of 59.2%.

1-(2-(1-Morpholinyl)acetyl)-rockmilanol (10u). White solid; mp 97-98°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.16-7.05 (7H, m ),6.64(2H,t,J＝6.0Hz),6.24(1H,d,J＝1.9Hz),6.03(1H,d,J＝1.9Hz),5.85(1H,d,J＝3.7Hz), 4.11(1H,dd,J=13.9, 6.1Hz), 3.84(3H,s), 3.76(3H,s), 3.67(3H,s), 3.66(4H,m), 3.05(1H,d,J= 16.8 Hz), 2.93 (1H, d, J=16.8 Hz), 2.89–2.85 (1H, m), 2.50–2.31 (5H, m), 2.20 (1H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 168.8, 163.7, 161.0, 158.5, 157.9, 138.4, 128.6, 128.0, 128.0, 127.8, 127.8, 127.1, 127.1, 126.2, 126.2, 112.6, 106.5, 103.0, 93.2, 91.7, 88.1, 79.3, 6.7, 58.1 55.5, 55.4, 54.9, 53.7, 52.9, 52.9, 35.6, 35.6; ESIMS m/z 562.2[M+ H] ⁺ ; HRESIMS m/z 562.2424[M+H] ⁺ (Calcd.for C ₃₂ H ₃₆ O ₈ N, 562.2435).

Example 22: Preparation of target compound 10v

The steps were the same as those in Example 1, except that the benzoic acid was replaced with piperidine to prepare the target compound 10v with a yield of 54.6%.

1-(2-(1-Piperidinyl)dioctylaminoacetyl)-rockmilanol (10v). White solid; mp 85-86°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.14-7.06 (7H, m), 6.64 (2H, m), 6.23 (1H, d, J=1.9 Hz), 6.03 (1H, d, J=1.9 Hz), 5.84 (1H, dd, J=4.8, 1.3 Hz) ,4.11(1H,dd,J＝13.8,6.1Hz),3.84(3H,s),3.75(3H,s),3.67(3H,s),3.03(1H,dd,J＝16.7Hz),2.88( 1H, dd, J=16.7 Hz), 2.84 (1H, td, J=14.0, 5.2 Hz), 2.43-2.29 (5H, m), 1.72-1.40 (6H, m); ¹³ C NMR (100 MHz, CDCl ₃ )δ: 169.4, 163.6, 161.0, 158.5, 158.0, 138.5, 128.7, 128.7, 128.0, 128.0, 127.8, 127.8, 127.2, 127.2, 126.2, 112.6, 106.7, 103.0, 93.3, 91.7, 5.8, 5.9 , 55.5, 55.4, 55.0, 53.8, 53.7, 35.7, 25.8, 25.8, 23.8; ESIMS m/z 560.2[M+H] ⁺ ; HRESIMS m/z 560.2650[M+H] ⁺ (Calcd.for C ₃₃ H ₃₈ O ₇ N, 560.2643).

Example 23: Preparation of target compound 10w

The steps were the same as those in Example 1, except that the benzoic acid was replaced with N-methylpiperazine to prepare the target compound 10w with a yield of 55.2%.

1-(2-(4-Methylpiperazin-1-yl)acetyl)-rockmilanol (10w). White solid; mp 88-89°C; ¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.15– 7.04(7H,m),6.63(2H,d,J＝8.9Hz),6.22(1H,d,J＝1.9Hz),6.02(1H,d,J＝1.9Hz),5.84(1H,d,J =3.8Hz), 4.11–4.04(1H,m), 3.83(3H,s), 3.75(3H,s), 3.67(3H,s), 3.03(1H,d,J=16.8Hz), 2.85(1H ,d,J＝16.8Hz),2.84(1H,td,J＝3.8Hz),2.43-2.41(8H,m),2.30(1H,ddd,J＝13.1,6.5,1.2Hz),2.29(3H, s); ¹³ C NMR (100 MHz, CDCl ₃ ) δ: 168.9, 163.7, 160.9, 158.5, 158.0, 138.4, 128.7, 128.7, 128.0, 128.0, 127.8, 127.8, 127.2, 126.2, 112.6, 112.6, 106.7, 112.6, 106.7 93.2, 91.7, 88.2, 79.3, 59.0, 55.5, 55.4, 55.4, 55.0, 54.7, 53.7, 53.4, 52.7, 45.9, 35.6; ESIMS m/z 574.2[M+H] ⁺ ; HRESIMS m/z 575.2751[M+ H] ⁺ (Calcd. for C ₃₃ H ₃₉ O ₇ N ₂ ,575.2752).

Test Example 1: In vitro antitumor activity screening

Cell lines: human erythroleukemia cell line (HEL), breast cancer cells (MDA-MB-231), human colon cancer cells (HCT116).

Experimental principle: MTT colorimetric method was used for detection. The full name of MTT is 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide, which is a yellow dye. Succinate dehydrogenase in the mitochondria of living cells can reduce MTT, and under the action of cytochrome C, it can generate blue (or blue-purple) water-insoluble formazan (Formazan), which is measured with a microplate reader at 490nm Formazan content. In general, the amount of formazan produced is positively correlated with the number of living cells, so the number of living cells can be inferred from the optical density OD value.

Experimental method: cells in logarithmic growth phase were seeded in 96-well plate with 6×10 ³ cells/well, placed in a 37 ℃, 5% CO ₂ cell incubator for 16 h, and observed that when the cells were in good condition and reached the logarithmic growth phase, Compounds with different concentration gradients were added to continue the culture, and adriamycin was used as a positive control. After culturing for 72 h at 37°C and 5% CO ₂ , 20 μL/well MTT (5 mg/mL) was added, and the culture was continued for 4 h. The 96-well plate was centrifuged at 2500 r/min for 15 min, the supernatant was discarded, 160 μL/well DMSO was added, and the cells were placed in Shake at 37°C in the dark for 15 min at low speed to fully dissolve the precipitate.

Colorimetry: OD value was determined using a multi-function microplate reader. The absorption wavelength was adjusted to 490 nm, and the light absorption value of each well of the 96-well plate was measured and recorded. The inhibition rate calculation for each well was then performed, followed by the determination of the _IC50 value.

Experimental results: The 20 synthesized compounds were screened and evaluated for their antitumor activity in vitro on three cell lines, HEL, MDA-MB-231 and HCT116. Doxorubicin was used as a positive control group, and the results were expressed as _IC50 values (see Table 1). ). In HCT116 tumor cell screening, the results showed that 13 compounds (compounds 10g, 10h, 10j, 10k, 10l, 10m, 10p, 10q, 10r, 10t, 10u, 10u and 10w) had better activity than the positive control doxorubicin antitumor activity.

Table 1 Activity screening data of 24 derivatives

Test Example 2: Study on the mechanism of anti-tumor action of the highly active representative compound 10r

1. Study on the effect of compound 10r on cell cycle

Different concentrations of compound 10r (0.05, 0.1 and 0.2 μM) were used to treat HCT116 tumor cells according to the following experimental procedures, and the results showed that this compound could dose-dependently block HCT116 tumor cells in G1 phase.

The specific experimental steps for the effect of compound 10r on the cycle of HCT116 tumor cells are as follows:

(1) Processing samples and collecting cells: take the cells in the logarithmic growth phase and inoculate them in a 60mm culture dish at a concentration of 3.5×10 ⁴ /ml, 3 ml per dish, and place them at 37°C, 5% CO ₂ cells After 24 hours in the incubator, the cells were divided into different time groups, each time group was divided into a control group and a sample intervention group with different concentrations, and the control group was 0.1% DMSO. The cells in each group were placed in a 37°C, 5% CO ₂ cell incubator for 48 h and collected;

(2) Fixation: collect the cell culture medium, wash the cells with pre-cooled PBS, add 1.5 mL of EDTA-free trypsin, digest for about 3 minutes, observe the cells begin to loosen, and gently blow the cells with a micropipette to collect the cells Suspension, wash the dish with 1.5mL PBS and collect. Centrifuge at 1000 r/min for 3 min with a centrifuge, discard the supernatant, wash the cells with PBS, continue at 1000 r/min, centrifuge for 3 min, discard the supernatant, add 500 μL of pre-cooled 75% ethanol, -20°C overnight fixation;

(3) Staining: before staining, wash twice with PBS, discard the supernatant, then add 500 μL of staining solution (0.05% TRITON, 0.5% RNAase and 5% PI) prepared in PBS, and protect from light Incubate at 37°C for 30min;

(4) Put on machine: use a centrifuge to spin the stained cells at 1000 r/min, centrifuge for 3 min, discard the supernatant, resuspend with 300 μl PBS, filter and put on machine.

The experimental results are shown in Figure 1 and Figure 2: Figure 1 shows the effect of compound 10r on the cycle of HCT116 tumor cells for 48 hours; 10r prevented HCT116 cell cycle, and cells were treated with different concentrations of compounds (0.05 μM, 0.1 μM and 0.2 μM)48 Hour. Figure 2 is a graph showing the percentage of compound 10r on HCT116 tumor cells at different stages; each experiment was repeated at least three times, *P<0.05, **P<0.01, the results show that this compound can dose-dependently block HCT116 tumor cells in G1 Expect.

2. Study on the effect of compound 10r on apoptosis

Different concentrations of compound 10r (0.05μM, 0.1μM and 0.2μM) were used to treat HCT116 tumor cells according to the following experimental procedures. The results showed that this compound could induce apoptosis of HCT116 tumor cells in a dose-dependent manner.

Compounds induce apoptosis in cells

(1) Processing samples and collecting cells: Take cells in logarithmic growth phase, inoculate 3.0×10 ⁵ /ml in 60mm petri dishes, 3 ml per dish, and place them in a 37°C, 5% CO ₂ cell incubator After 24 hours, the cells were divided into control group and 10r sample group. The control group was 0.1% DMSO group, and the sample groups were 0.05 μM, 0.1 μM and 0.2 μM, respectively. The cells in each group were incubated in a cell incubator for 48 hours and collected. each group of cells;

(2) Collect the cell culture medium, wash the cells with PBS, add 1.5 mL of EDTA-free trypsin, digest for 3 minutes, observe the cells begin to loosen, blow the cells gently, collect the cell suspension, wash the culture dish with 1.5 mL of PBS and collect . Centrifuge at 1000 r/min for 3 min, discard the supernatant, wash the cells with PBS, repeat twice;

(3) Staining: add 50 μl Binding Buffer to resuspend the cells, transfer them to the flow tube, add propidium iodide (PI) 2.5 μl/tube, phospholipid-binding protein V (Annexin V-FITC) to each tube in the drug-adding group ) 2.5 μl/tube, gently mix the cells, and incubate at room temperature for 20 min in the dark;

(4) On the machine: Centrifuge the stained cells at 1000 r/min for 3 min, discard the supernatant, add 50 μl of 1×Binding Buffer to each tube, and perform flow cytometry analysis as soon as possible;

The experimental results are shown in Figure 3 and Figure 4: Figure 3 is a graph of the apoptosis of HCT116 tumor cells induced by compound 10r; flow cytometry analysis of different concentrations of compound 10r (0.05μM, 0.1μM and 0.2M); Figure 4 is compound 10r induced tumor cells Histogram of apoptosis; each experiment was repeated at least three times, *P<0.05, **P<0.01 compared with the control group. The results show that this compound can induce apoptosis of HCT116 tumor cells in a dose-dependent manner.

3. The effect of compound 10r on the formation of HCT116 cloning spheres

HCT116 cells in logarithmic growth phase were seeded in 6-well plates at 500/well, placed in a 37°C, 5% CO ₂ cell incubator overnight, and replaced with fresh medium. , 0.05 and 0.1 μM) intervention group, and the control group was 0.1% DMSO group. The cells were cultured at 37 °C with 5% _CO2 , and the medium was changed every three days for 15 consecutive days. After washing three times with PBS, 500 μL of methanol was added for fixation for 30 min, washed twice with PBS, stained with crystal violet for 20 min, washed three times with PBS, dried and photographed.

The experimental results are shown in Figure 5 and Figure 6: Figure 5 shows the effect of compound 10r on the formation of HCT116 clone spheres; Figure 6 shows the columnar statistics of the number of clone spheres formed after HCT116 was treated with compound 10r (0.05μM, 0.1μM and 0.2μM) for 15 days Figure; each experiment was repeated at least three times, *P<0.05, **P<0.01 compared with the control group. The results showed that this compound could inhibit the formation of HCT116 clonal spheres in a dose-dependent manner.

Previous experiments showed that 10r could inhibit the proliferation of HCT116 cells, induce apoptosis in a dose-dependent manner, and arrest cells in G1 phase. In order to further confirm the inhibitory effect of 10r on cell proliferation, HCT116 was taken as the research object to explore the effect of 10r on the formation of clonal spheres. The results are shown in 5 and 6, 10r significantly inhibited HCT116 clonal sphere formation in a dose-dependent manner. The cloning ability induced by 10r (0.025 μM) was almost completely inhibited compared to the control group.

4. The effect of compound 10r on the expression of related apoptosis proteins, cyclins, MAPK signaling pathway and key proteins of Wnt signaling pathway

HCT116 cells were seeded at 1×10 ⁶ /dish in a 100 mm petri dish and cultured overnight. Different concentrations of 10r (0.05, 0.1 and 0.2μM) were used to treat cells, and 0.1% DMSO was added to the control group. Cells were collected and lysed. The total protein obtained was quantified by BCA, and the protein was separated by 10% SDS-PAGE and transferred to PVDF membrane. 5% skim milk for blocking, 4°C primary antibodies (c-Myc, G1/S-specific cyclin-E, G1/S-specific cyclin-D, cyclin-dependent kinase 4, cyclin-dependent kinase 6. Full-length-PARP1, split-PARP1, pro-caspase-3, split-pro-caspase--3, pro-caspase-9, split-pro-caspase-9, Bcl-2, p-ERK, ERK, p-JNK, JNK, p-P38, P38, β-catenin, Axin-2, GSK-3β, TCF-4, β-actin and glyceraldehyde phosphate dehydrogenase) incubation After overnight incubation with secondary antibody at room temperature for 2 h. Use the Odyssey Platform for detection.

The experimental results are shown in Figure 7 and Figure 8: Figure 7 is a graph showing the effect of compound 10r on the expression of related apoptotic proteins; . Figure 8 is a graph showing the effect of compound 10r on the expression of related cyclins;

Based on the previous experimental results, Western bolt was used to analyze the effect of 10r on apoptosis and cell cycle-related proteins, and to further explore the molecular mechanism of 10r-induced apoptosis and cell cycle arrest in HCT116 cells. As shown in Figure 7, 10r induced specific cleavage of the apoptotic proteins PARP, procaspase-3 and procaspase-9 in HCT116 in a dose-dependent manner. In addition, with the increase of 10r concentration, the expression of anti-apoptotic protein Bcl-2 was down-regulated, indicating that 10r may induce apoptosis through mitochondrial pathway. 10r induced HCT116 to arrest in the G1 phase of the cell cycle, and analyzed the effect of 10r on the expression of cyclins and related protein kinases in the G1 phase. As shown in Figure 8, 10r (0.1 μM) significantly reduced c-Myc, G1/S-specific cyclin-D, G1/S-specific cyclin-E, cyclin-dependent kinase 4 and cyclin-dependent Kinase 6 expression, from which it can be speculated that 10r may be mediated by degradation of c-Myc, G1/S-specific cyclin-D, G1/S-specific cyclin-E, cyclin-dependent kinase 4 and cell cycle Kinase 6-dependent, whereby the cyclin-CDK complex cannot properly form, resulting in HCT116 arrest in G1.

The experimental results are shown in Figure 9 and Figure 10: Figure 9 shows the effect of compound 10r on the expression of proteins related to the mitogen-activated protein kinase (MAPK) signaling pathway; Figure 10 is a graph showing the effect of compound 10r on the expression of key proteins in the Wnt signaling pathway.

Mitogen-activated protein kinase (MAPK) and Wnt family, as key signaling molecules regulating cell proliferation, homeostasis and development, are involved in a series of cellular physiological activities such as cell growth, differentiation and apoptosis. Western bolt was used to detect the effects of 10r on MAPK and wnt signaling pathways. As shown in Figure 9, 10r significantly up-regulated the expression levels of p-P38 and p-JNK, and down-regulated the expression level of p-ERK in a dose-dependent manner. As shown in Figure 10, after 10r treatment, the expressions of β-catenin, Axin-2, GSK-3β and TCF-4 were significantly decreased in a dose-dependent manner.

The above are only preferred embodiments of the present invention, and do not limit the present invention in any form, and any simple modifications made to the above embodiments according to the technical essence of the present invention without departing from the technical solution content of the present invention, Equivalent changes and modifications still fall within the scope of the technical solutions of the present invention.

Claims (4)

1. A hydroxy derivative of loxagliflorin having the following structural formula (I):

wherein: r is a benzoate group, a p-fluorobenzoate group, a p-methoxybenzoate group, a p-bromobenzoate group, a 4, 5-dimethoxy-2-nitro-benzoate group, a p-nitrocinnamate group, a 2-furoate group, a 2-picolinate group, a 6-chloronicotinate group, a butyrate group, a decanoate group, a dimethylamino group, a diethylamino group, a dibutylamino group, a dioctylamino group, a benzylamino group, a 3, 4-dimethoxyphenethylamino group, a 1-tetrahydropyrrolyl group, a 1-imidazolyl group, a 1-pyrazolyl group, a 1-morpholinyl group, a 1-piperidinyl group or a 4-methylpiperazin-1-yl group.

2. A preparation method of a rocomimol hydroxyl derivative comprises the following steps:

(1)

synthesis of (2)

Weighing 20g

Putting the mixture into a 500ml round bottom flask, adding 51g of anhydrous potassium carbonate and 250ml of acetone into the flask as solvents, slowly adding 24.5ml of dimethyl sulfate while stirring, and putting the system into an oil bath kettle for heating and refluxing at 70 ℃ for 72 hours. TLC tracing reaction, after the reaction is finished, cooling to room temperature, adjusting pH to 11 with ammonia water, filtering to remove precipitate, washing filtrate with saturated saline solution, drying with anhydrous sodium sulfate, spin-drying partial solvent, adding silica gel for sample mixing, purifying with flash silica gel column, and obtaining light yellow solid with acetone of 8:2

Weighing 700mg

Adding a mixed solvent of 80mL of dichloromethane and 60mL of acetone, and slowly adding 140mL of potassium hydrogen monosulfate complex salt aqueous solution with the concentration of 11.6g/mL to obtain

(2)

And with

Synthesis of the mixture

Weighing 800mg of

Adding 40ml of acetonitrile, 30ml of methanol and 12.6 equivalents of trans-methyl cinnamate 5.0g, irradiating by using a xenon lamp for strong light, and reacting for 17 hours to obtain a product

And

a mixture of (a);

(3)

synthesis of (2)

Weighing 1g of

And

30mL of methanol and 10mL of a 0.5M sodium methoxide solution in methanol were added to the mixture, and the mixture was refluxed at 70 ℃ for 4 hours to obtain

(4)

Synthesis of (2)

Weighing 2g

100mL of DMSO and 170mg of lithium chloride were added thereto, and the mixture was stirred at 100 ℃ for 8 hours to obtain

(5)

Synthesis of (2)

3.27g of sodium triacetoxyborohydride are weighed into 200mL of acetonitrile and 1.2mL of glacial acetic acid, and 700mg of sodium triacetoxyborohydride are slowly added

Stirring at 40 deg.C for 8h to obtain

(6)

Synthesis of (2)

Weighing 30mg

Placed in a 25ml round bottom flask, 2ml DCM was added as solvent and 28.7. mu.l Et was added₃Stirring N, 11 mu l of chloroacetyl chloride and a catalytic amount of DMAP at room temperature for 10h to obtain

Namely 1-chloroacetyl-4' -demethoxy-clomipraminol;

(7)

synthesis of series of derivatives

Weighing 30mg of

5ml of DMF solution, 16.3mg of potassium carbonate (0.118mmol) and 1.5 equivalents of acid (0.085mmol) are added and the mixture is heated in a thermostatic oil bath at 70 ℃ for 12 hours to give the series of derivatives

Wherein: r is a benzoate group, a p-fluorobenzoate group, a p-methoxybenzoate group, a p-bromobenzoate group, a 4, 5-dimethoxy-2-nitro-benzoate group, a p-nitrocinnamate group, a 2-furoate group, a 2-picolinate group, a 6-chloronicotinate group, a butyrate group, a decanoate group, a dimethylamino group, a diethylamino group, a dibutylamino group, a dioctylamino group, a benzylamino group, a 3, 4-dimethoxyphenethylamino group, a 1-tetrahydropyrrolyl group, a 1-imidazolyl group, a 1-pyrazolyl group, a 1-morpholinyl group, a 1-piperidinyl group or a 4-methylpiperazin-1-yl group.

3. Application of a loxagliflorol hydroxyl derivative in preparation of medicaments for resisting colorectal cancer, leukemia and breast cancer.

4. An application of a loxagliflorin hydroxyl derivative in the preparation of MAPK signal channel and Wnt signal channel inhibitor drugs.

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