CN113004241A

CN113004241A - 11, 20-dicarbonyl economic oridonin 14-O-benzoic acid esterified derivative and preparation method and application thereof

Info

Publication number: CN113004241A
Application number: CN202110242717.7A
Authority: CN
Inventors: 可钰; 李富欣; 王妮; 贾小苹; 赵梦圆
Original assignee: Henan Tobacco Company Jiyuan Co; Zhengzhou University
Current assignee: Henan Tobacco Company Jiyuan Co; Zhengzhou University
Priority date: 2021-03-05
Filing date: 2021-03-05
Publication date: 2021-06-22
Anticipated expiration: 2041-03-05
Also published as: CN113004241B

Abstract

The invention relates to the field of natural products and pharmaceutical chemistry, and discloses an 11, 20-dicarbonyl economic oridonin 14-O-benzoic acid esterified derivative, a preparation method and application thereof. The preparation method comprises the following steps: the oridonin (JOA) is used as a starting material and is oxidized to obtain 11, 20-dicarbonyl oridonin, and then benzoic acid or substituted benzoic acid is subjected to esterification reaction with 14-OH of the oridonin to obtain a series of 11, 20-dicarbonyl oridonin 14-O-benzoic acid esterified derivatives on the premise of not damaging the active center alpha-methylene cyclopentanone. The compounds have good antitumor activity, and can be used for preparing anticancer drugs for treating esophageal cancer, gastric cancer, liver cancer, breast cancer, pancreatic cancer and other diseases. The general structural formula is as follows:

Description

11, 20-dicarbonyl economic oridonin 14-O-benzoic acid esterified derivative and preparation method and application thereof

Technical Field

The invention relates to the field of natural products and pharmaceutical chemistry, in particular to an 11, 20-dicarbonyl economic oridonin 14-O-benzoic acid esterified derivative, a preparation method and application thereof.

Background

Rabdosia rubescens is plant of Rabdosia of Labiatae, and its application part is dried aerial part, and its effect is clearing heat and detoxicating, promoting blood circulation and relieving pain. A large number of researches show that the main anti-tumor active component in rabdosia rubescens is a diterpenoid compound, and the ent-kaurene diterpenoid is a natural compound with good anti-tumor effect, and documents report that the anti-tumor effect active center of the structure mainly depends on the molecular configuration and the cyclopentanone structure conjugated with exocyclic methylene, such as oridonin, rabdosia rubescens B, rabdosia leptandra and the like, and the anti-tumor effect active center of the structure has the anti-proliferation effect on various tumor cells in vitro, such as cancer cells of esophagus cancer, stomach cancer, breast cancer, cervical cancer, pancreatic cancer and the like.

The inventor separates the oridonin from the rabdosia rubescens ethanol extract by a series of methods in the earlier stage, and oxidizes the oridonin by using a Jones reagent to obtain the required lead compound 11, 20-dicarbonyl oridonin. The in vitro anti-tumor activity shows that the compound has better anti-proliferation effect on tumor cells and wider anti-tumor spectrum. Although most of the compounds have good anti-tumor effect, the defects of poor water solubility, poor stability, difficult oral absorption and the like generally exist, and the clinical application of the compounds is limited. The structural modification and reconstruction of the compounds are carried out to overcome the defects, and the problem of seeking a target compound with better pharmacy is the problem which needs to be solved urgently at present.

Disclosure of Invention

The invention aims to provide a series of Jiyuan rubescensine A derivatives, improve the anti-tumor effect, increase the bioavailability and increase the clinical application possibility of the Jiyuan rubescensine A.

The invention also aims to provide a preparation method and application thereof in preparing antitumor drugs.

In order to realize the purpose of the invention, hydroxyl at 11-position and 20-position of the oridonin from Jiyuan is oxidized into carbonyl, and then 14-OH of the oridonin is esterified with benzoic acid to obtain a series of derivatives, thus maintaining or enhancing the anti-tumor effect and simultaneously increasing the stability and bioavailability of the oridonin.

The structural general formula of the benzoic acid series derivative esterified by the 11, 20-dicarbonyl economic oridonin 14-OH is as follows:

wherein R is mono-substituted, di-substituted or polysubstituent on benzene ring, and R is selected from hydrogen; a methoxy group; halogen; a nitro group; an amino group; a sulfonic acid group; a linear or branched alkyl group; straight chain alkyl substituted with halogen; a trifluoromethyl group.

R is preferably: hydrogen; c1-6 straight chain alkyl; c1-6 straight chain alkyl substituted with halogen; halogen; a nitro group; a methoxy group; a trifluoromethyl group.

R is preferably: hydrogen; a methoxy group; a methyl group; chlorine; bromine; fluorine; a trifluoromethyl group; iodine; a nitro group; a bromomethyl group.

The following groups after R substitution are more preferred:

the novel 11, 20-dicarbonyl economic oridonin 14-OH esterified benzoic acid series derivative is obtained by the following synthetic route:

1. dissolving separated and purified oridonin (JOA) in acetone, adding Jones reagent for reaction, adding isopropanol after the reaction is finished to quench the reaction, and purifying by column chromatography to obtain mother nucleus 11, 20-dicarbonyl oridonin.

2. Dissolving the obtained lead compound 11, 20-dicarbonyl economic oridonin in dichloromethane, adding corresponding benzoic acid, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) and 4-Dimethylaminopyridine (DMAP) as catalysts under stirring for reaction, and purifying by column chromatography to obtain the target compound.

The invention has the innovation points and advantages that: the invention selects oridonin (JOA) as a lead compound, obtains the required derivative through oxidation and esterification processes, has increased stability and better anti-tumor effect, can be used as a potential medicament for treating esophageal cancer, gastric cancer, liver cancer, breast cancer, pancreatic cancer and the like, and has important theoretical significance and practical application value for developing novel anti-tumor medicaments.

Detailed Description

The present invention is further illustrated by the following specific examples, but it should be noted that the scope of the present invention is not limited in any way by these examples.

Example 1:

weighing JOA 300mg, adding 11mL of acetone to completely dissolve the acetone, adding 0.75mL of Jones reagent (6mL of acetone for dilution) under the stirring condition, reacting for 20min, monitoring the reaction completion by a dot plate, adding a certain amount of isopropanol to quench the reaction, performing reduced pressure spin drying after 20min, adding 30mL of ethyl acetate to dilute the reaction system, washing the reaction system with saturated NaCl solution for three times, performing back extraction on the water phase once with ethyl acetate, combining organic phases, drying with anhydrous magnesium sulfate, concentrating, and performing column chromatography purification to obtain 270mg of a target compound 11, 20-dicarbonyl economic oridonin with a yield of 74%.¹H NMR(400MHz,DMSO-d₆)δ6.14(s,1H),6.08(d,J＝2.9Hz,1H),5.79(s,1H),4.79(dd,J＝3.9,1.7Hz,1H),3.74–3.71(m,1H),3.10(d,J＝8.3Hz,1H),2.96(s,1H),2.92–2.83(m,2H),2.66(dd,J＝16.3,8.4Hz,1H),2.55(s,1H),1.76(ddd,J＝14.4,7.6,4.0Hz,1H),1.59(dp,J＝12.6,5.1,4.3Hz,2H),1.36(d,J＝11.6Hz,2H),1.09(td,J＝11.4,10.0,5.2Hz,1H),0.98(td,J＝14.0,4.4Hz,1H),0.81(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ204.50,200.41,174.32,148.15,121.28,72.87,70.54,58.52,57.72,46.51,45.35,42.97,41.08,33.81,30.56,28.28,23.36,19.10,18.21.HR-MS(ESI):Calculated for C₂₀H₂₄O_5.[M+H]⁺:345.1702,found:345.1688.

Example 2:

compound 1a

Weighing 200mg of 11, 20-dicarbonyl economic oridonin, completely dissolving the oridonin in 10mL of dichloromethane, adding 92mg of benzoic acid under stirring, and sequentially adding 144mg of catalyst 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) and 9mg of 4-Dimethylaminopyridine (DMAP). After the reaction is monitored by a thin-layer chromatography plate, 30mL of dichloromethane is added to dilute the reaction system, the reaction system is washed three times by saturated sodium bicarbonate solution, water layers are combined and back-extracted once, organic phases are combined, dried by anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain a white solid product, and the yield is 85%.¹H NMR(400MHz,DMSO-d₆)δ7.62(d,J＝7.8Hz,2H),7.44(d,J＝7.5Hz,1H),7.29(t,J＝7.5Hz,2H),6.04(s,1H),5.68(s,1H),4.79(s,1H),4.60(d,J＝3.5Hz,1H),3.10(s,2H),3.07(s,1H),2.66(d,J＝13.8Hz,2H),2.59(t,J＝8.3Hz,1H),2.26(s,1H),1.60–1.53(m,1H),1.46–1.36(m,2H),1.14(d,J＝11.8Hz,2H),0.86(d,J＝8.2Hz,1H),0.57(s,3H),0.47(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.35,198.48,173.92,146.12,134.03,129.26,128.93,128.34,122.99,74.02,72.56,58.91,56.40,46.55,45.35,43.21,38.58,33.83,30.50,28.17,23.16,18.94,18.16.HR-MS(ESI):Calculated for C₂₇H₂₈O_6.[M+NH₄]⁺:466.2230.found:466.2227.

Example 3:

compound 2a

The same procedures used in example 2 were repeated except for using 115mg of 2-methoxybenzoic acid in place of benzoic acid to give a white solid product in a yield of 47%.¹H NMR(400MHz,DMSO-d₆)δ7.61(dd,J＝7.9,1.8Hz,1H),7.58–7.55(m,1H),7.16–7.13(m,1H),7.04–7.01(m,1H),6.25(s,1H),5.91(s,1H),4.99(d,J＝1.1Hz,1H),4.83(dd,J＝4.1,1.8Hz,1H),3.77(d,J＝2.5Hz,3H),3.46(d,J＝8.4Hz,1H),3.28(s,1H),2.92(ddd,J＝13.1,8.0,2.6Hz,2H),2.81(d,J＝8.2Hz,1H),2.71(d,J＝16.4Hz,1H),1.89–1.84(m,1H),1.69–1.65(m,1H),1.41(d,J＝3.9Hz,1H),1.37(s,1H),1.23(s,1H),1.11(d,J＝3.8Hz,1H),1.09–1.07(m,1H),0.83(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.48,198.47,174.02,164.49,158.56,146.22,134.56,131.10,122.73,120.25,118.25,112.78,73.80,72.65,58.81,56.30,55.71,46.55,45.32,43.16,39.76,38.55,33.83,30.49,28.16,23.13,18.97,18.15.HR-MS(ESI):Calculated for C₂₈H₃₀O_7.[M+NH₄]⁺:496.2335.found:496.2328.

Example 4

Compound 2b

The same procedures used in example 2 were repeated except for using 115mg of 3-methoxybenzoic acid in place of benzoic acid to give a white solid product in a yield of 75%.¹H NMR(400MHz,DMSO-d₆)δ7.44(q,J＝2.1,1.6Hz,2H),7.33(t,J＝2.1Hz,1H),7.29–7.26(m,1H),6.29(s,1H),5.92(s,1H),5.03(d,J＝5.3Hz,1H),4.84–4.80(m,1H),3.80(s,1H),3.30(s,1H),2.92(dt,J＝3.9,2.0Hz,1H),2.88(d,J＝5.6Hz,1H),2.82(d,J＝8.2Hz,1H),2.72(d,J＝16.4Hz,1H),1.85–1.80(m,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.40(s,1H),1.37(s,1H),1.24(d,J＝11.1Hz,1H),1.11(d,J＝3.2Hz,1H),1.08(d,J＝4.3Hz,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.39,198.47,173.97,164.13,159.29,146.07,130.20,129.64,121.39,119.62,114.32,74.06,72.58,58.89,56.41,55.33,46.52,45.33,43.20,39.73,38.57,33.82,30.48,28.16,23.16,18.93,18.14.HR-MS(ESI):Calculated for C₂₈H₃₀O_7.[M+NH₄]⁺:496.2335.found:496.2336.

Example 5

Compound 2c

The same procedures used in example 2 were repeated except for using 115mg of 4-methoxybenzoic acid in place of benzoic acid to give a white solid product in a yield of 63%.¹H NMR(400MHz,DMSO-d₆)δ7.81(d,J＝8.8Hz,2H),7.06(d,J＝8.7Hz,2H),6.27(s,1H),5.92(s,1H),4.98(s,1H),4.83–4.79(m,1H),3.47(d,J＝8.0Hz,1H),3.33(s,3H),2.90(d,J＝14.1Hz,2H),2.81(d,J＝8.3Hz,1H),2.71(d,J＝16.4Hz,1H),1.85–1.79(m,1H),1.73–1.62(m,2H),1.59(d,J＝14.0Hz,1H),1.40(s,1H),1.37(s,1H),1.12(d,J＝12.5Hz,1H),1.09–1.04(m,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.42,198.57,173.93,163.70,146.21,131.47,122.91,120.43,114.25,73.62,72.58,58.87,56.38,55.59,46.54,45.33,43.19,38.62,33.84,30.51,28.17,23.16,18.95,18.15.HR-MS(ESI):Calculated for C₂₈H₃₀O_7.[M+NH₄]⁺:496.2335.found:496.2331.

Example 6

Compound 2d

The same procedure used in example 2 was repeated except for using 137mg of 2, 3-dimethoxybenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 63%.¹H NMR(400MHz,DMSO-d₆)δ7.30(dd,J＝5.8,3.9Hz,1H),7.18–7.15(m,2H),6.26(s,1H),5.94(s,1H),5.05(d,J＝1.2Hz,1H),4.79(dt,J＝4.5,2.2Hz,1H),3.83(s,3H),3.64(d,J＝3.9Hz,3H),3.48(d,J＝8.1Hz,1H),3.30(s,1H),2.94–2.88(m,2H),2.82(d,J＝8.3Hz,1H),2.72(d,J＝16.5Hz,1H),1.87–1.81(m,1H),1.67(d,J＝3.0Hz,1H),1.40(d,J＝4.6Hz,1H),1.37(s,1H),1.26(s,1H),1.11(d,J＝3.9Hz,1H),1.08(d,J＝4.5Hz,1H),0.82(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.39,198.53,173.97,164.47,153.14,148.02,146.20,124.35,124.10,122.95,121.58,117.18,74.11,72.62,60.75,58.89,56.30,56.02,46.57,45.30,43.17,39.75,38.58,33.82,30.49,28.15,23.12,18.96,18.14.HR-MS(ESI):Calculated for C₂₉H₃₂O_8.[M+NH₄]⁺:526.2441.found:526.2435.

Example 7

Compound 2e

The same procedure used in example 2 was repeated except for using 137mg of 3, 4-dimethoxybenzoic acid in place of benzoic acid to give a white solid product with a yield of 41%.¹H NMR(400MHz,DMSO-d₆)δ7.45(dd,J＝8.4,2.0Hz,1H),7.34(d,J＝2.1Hz,1H),7.07(d,J＝8.6Hz,1H),6.28(s,1H),5.92(s,1H),4.99(d,J＝1.3Hz,1H),4.83–4.80(m,1H),3.83(s,3H),3.78(s,3H),3.46(d,J＝2.0Hz,1H),3.30(s,1H),2.92(s,1H),2.88(s,1H),2.86(s,1H),2.75–2.72(m,1H),1.85–1.80(m,1H),1.68(dd,J＝10.5,7.6Hz,1H),1.38(s,1H),1.23(s,1H),1.09(d,J＝2.2Hz,1H),1.05(d,J＝2.2Hz,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.43,198.56,173.93,162.28,154.44,148.81,148.54,124.98,120.36,120.06,112.22,111.44,73.71,72.59,58.85,56.41,55.92,55.63,46.53,45.35,43.20,40.00,38.64,33.84,30.51,28.17,23.18,18.95,18.15.HR-MS(ESI):Calculated for C₂₉H₃₂O_8.[M+NH₄]⁺:526.2441.found:526.2440.

Example 8

Compound 2f

The same procedure used in example 2 was repeated except for using 160mg of 3,4, 5-trimethoxybenzoic acid in place of benzoic acid to give the product as a white solid in 82% yield.¹H NMR(400MHz,DMSO-d₆)δ7.11(s,2H),6.29(s,1H),5.94–5.91(m,1H),5.00(d,J＝1.2Hz,1H),4.85(dd,J＝4.1,1.7Hz,1H),3.80(s,6H),3.74(d,J＝2.0Hz,3H),3.31(s,1H),2.92(dd,J＝4.2,1.8Hz,1H),2.89(s,1H),2.82(d,J＝8.2Hz,1H),2.73(d,J＝16.4Hz,1H),1.82(dq,J＝11.4,3.6Hz,1H),1.71–1.66(m,1H),1.40(s,1H),1.37(s,1H),1.25(d,J＝10.6Hz,2H),1.11(s,1H),1.08(d,J＝4.0Hz,1H),0.82(s,3H),0.72(d,J＝3.6Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.45,198.47,174.00,163.92,152.77,146.12,142.53,123.32,122.94,106.74,74.10,72.58,60.20,58.85,56.42,56.00,46.50,45.32,43.20,39.73,38.60,33.82,31.09,30.48,29.77,28.15,23.23,18.94,18.15.HR-MS(ESI):Calculated for C₃₀H₃₄O_9.[M+NH₄]⁺:556.2547.found:556.2540.

Example 9

Compound 3a

The same operation as in example 2 was carried out using 102mg of 2-methylbenzoic acid instead of benzoic acid to give a product as a white solid in a yield of 64%.¹H NMR(400MHz,DMSO-d₆)δ7.71(dd,J＝7.8,1.5Hz,1H),7.54–7.50(m,1H),7.34(d,J＝3.2Hz,1H),7.33(d,J＝3.4Hz,1H),6.28(s,1H),5.94(s,1H),5.03(s,1H),4.82–4.78(m,1H),3.51(d,J＝8.0Hz,1H),3.31(s,1H),2.92(s,1H),2.90–2.87(m,1H),2.82(d,J＝8.3Hz,1H),2.72(d,J＝16.4Hz,1H),2.45(s,3H),1.84–1.79(m,1H),1.67(dd,J＝10.6,7.6Hz,2H),1.40(d,J＝2.1Hz,1H),1.38–1.36(m,1H),1.11(s,1H),1.08(d,J＝4.5Hz,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.41,198.58,173.94,165.16,146.29,139.76,132.95,131.87,130.25,127.82,126.21,122.98,73.93,72.60,58.84,56.35,46.51,45.28,43.16,39.95,38.61,33.83,30.51,28.16,23.18,21.21,18.97,18.15.HR-MS(ESI):Calculated for C₂₈H₃₀O_6.[M+NH₄]⁺:480.2368.found:480.2379.

Example 10

Compound 3b

The same operation as in example 2 was carried out using 102mg of 3-methylbenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 67%.¹H NMR(400MHz,DMSO-d₆)δ7.68(d,J＝1.9Hz,1H),7.65(d,J＝8.0Hz,1H),7.51(d,J＝7.6Hz,1H),7.42(d,J＝3.0Hz,1H),6.28(s,1H),5.92(s,1H),5.01(d,J＝1.1Hz,1H),4.87–4.84(m,1H),3.49(d,J＝8.0Hz,1H),3.31(s,1H),2.92(dd,J＝4.2,1.8Hz,1H),2.88(d,J＝6.4Hz,1H),2.82(d,J＝8.3Hz,1H),2.72(d,J＝16.5Hz,1H),2.36(s,3H),1.82(dq,J＝11.4,3.8Hz,1H),1.68(d,J＝3.1Hz,1H),1.64–1.57(m,1H),1.40(d,J＝2.4Hz,1H),1.38(s,1H),1.11(d,J＝3.5Hz,1H),1.09–1.04(m,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.38,198.52,173.94,164.47,146.12,138.42,134.67,129.52,128.82,128.33,126.48,123.01,74.02,72.55,58.92,56.38,46.57,45.35,43.22,38.56,33.84,30.51,28.17,23.18,20.73,18.94,18.15.HR-MS(ESI):Calculated for C₂₈H₃₀O_6.[M+NH₄]⁺:480.2368.found:480.2380.

Example 11

Compound 3c

The same operation as in example 2 was carried out using 102mg of 4-methylbenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 36%.¹H NMR(400MHz,DMSO-d₆)δ7.76(d,J＝7.9Hz,2H),7.34(d,J＝7.9Hz,2H),6.27(s,1H),5.92(s,1H),5.00(s,1H),4.85–4.80(m,1H),3.49(d,J＝8.1Hz,1H),2.92(t,J＝12.2Hz,2H),2.82(t,J＝8.3Hz,1H),2.71(d,J＝16.5Hz,1H),2.38(s,3H),1.86–1.79(m,1H),1.71–1.57(m,2H),1.44–1.36(m,2H),1.31–1.22(m,1H),1.08(dd,J＝13.8,9.9Hz,2H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.38,198.53,173.92,164.31,146.15,144.57,129.47(*2),129.31(*2),129.28,129.06,125.61,122.96,73.84,72.56,58.89,56.38,46.55,45.34,43.21,39.75,38.58,33.84,30.51,28.17,23.16,21.19,18.94,18.15.HR-MS(ESI):Calculated for C₂₈H₃₀O_6.[M+NH₄]⁺:480.2368.found:480.2381.

Example 12

Compound 4a

The same procedures used in example 2 were repeated except for using 118mg of 3-chlorobenzoic acid in place of benzoic acid to give a white solid product in a yield of 56%.¹H NMR(400MHz,DMSO-d₆)δ7.84–7.81(m,2H),7.79–7.76(m,1H),7.59(d,J＝8.0Hz,1H),6.30(s,1H),5.93(s,1H),5.05(s,1H),4.90–4.87(m,1H),3.52(d,J＝8.0Hz,1H),3.31(s,1H),2.92(dd,J＝4.1,1.7Hz,1H),2.90–2.87(m,1H),2.83(d,J＝8.2Hz,1H),2.73(d,J＝16.4Hz,1H),1.84–1.78(m,1H),1.71–1.66(m,1H),1.62(t,J＝3.6Hz,1H),1.40(s,1H),1.39–1.35(m,1H),1.11(d,J＝3.6Hz,1H),1.10–1.07(m,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.32,198.39,173.97,163.21,145.99,133.84,133.58,130.96,130.40,128.68,128.02,123.14,74.49,72.48,58.98,56.38,46.55,45.33,43.20,39.74,38.51,33.82,30.49,28.15,23.17,18.93,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇ClO_6.[M+NH₄]⁺:500.1840.found:500.1839.

Example 13

Compound 4b

The same operation as in example 2 was carried out using 118mg of 4-chlorobenzoic acid instead of benzoic acid to give a white solid product in 74% yield.¹H NMR(400MHz,DMSO-d₆)δ7.87(d,J＝8.2Hz,2H),7.61(d,J＝8.1Hz,2H),6.28(s,1H),5.93(s,1H),5.03(s,1H),4.85(d,J＝3.3Hz,1H),3.51(d,J＝8.1Hz,1H),2.92(t,J＝12.4Hz,2H),2.83(t,J＝8.3Hz,1H),2.72(d,J＝16.3Hz,1H),1.80(dq,J＝8.3,5.0Hz,1H),1.71–1.57(m,2H),1.44–1.35(m,2H),1.25(d,J＝10.4Hz,1H),1.08(dd,J＝15.6,11.3Hz,2H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.31,198.40,173.91,163.54,146.04,139.00,131.14(*2),129.12(*2),127.21,123.08,74.28,72.49,58.93,56.38,46.55,45.34,43.20,39.75,38.53,33.83,30.51,28.16,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇ClO_6.[M+NH₄]⁺:500.1840.found:500.1839.

Example 14

Compound 4c

The same procedure used in example 2 was repeated except for using 144mg of 2, 3-dichlorobenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 80%.¹H NMR(400MHz,DMSO-d₆)δ7.88(dd,J＝8.1,1.6Hz,1H),7.70(dd,J＝7.8,1.6Hz,1H),7.50(t,J＝8.0Hz,1H),6.30–6.25(m,1H),5.99–5.93(m,1H),5.08(d,J＝1.2Hz,1H),4.81(dt,J＝5.5,2.7Hz,1H),3.57(d,J＝8.4Hz,1H),3.32(s,1H),2.93–2.87(m,2H),2.86–2.82(m,1H),2.73(d,J＝16.4Hz,1H),1.81(ddd,J＝14.6,7.6,4.1Hz,1H),1.69–1.64(m,1H),1.41–1.36(m,2H),1.24(d,J＝3.8Hz,1H),1.10(d,J＝3.7Hz,1H),1.09–1.04(m,1H),0.81(s,3H),0.72(d,J＝2.8Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.19,198.18,173.90,163.25,146.00,133.99,133.20,131.38,129.67,129.55,128.70,123.18,75.23,72.47,58.92,56.32,46.50,45.28,43.17,39.76,38.49,33.82,30.50,28.14,23.13,18.97,18.15.HR-MS(ESI):Calculated for C₂₇H₂₆Cl₂O_6.[M+NH₄]⁺:534.1450.found:534.1442.

Example 15

Compound 4d

The same procedure used in example 2 was repeated except for using 144mg of 2, 6-dichlorobenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 78%.¹H NMR(400MHz,DMSO-d₆)δ7.59(s,2H),7.58(s,1H),6.26(s,1H),5.99(s,1H),5.08(s,1H),4.75–4.73(m,1H),3.57(d,J＝8.3Hz,1H),3.34(s,2H),2.95–2.91(m,1H),2.88(d,J＝8.3Hz,1H),2.86–2.80(m,1H),2.75(d,J＝16.5Hz,1H),1.87–1.81(m,1H),1.65(t,J＝2.9Hz,1H),1.39(d,J＝3.6Hz,1H),1.37(s,1H),1.23(s,1H),1.12–1.07(m,2H),0.81(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.09,197.96,173.92,162.83,145.77,132.91,131.48,130.40(*2),128.44(*2),123.42,75.59,72.47,58.89,56.17,46.37,45.25,43.18,39.73,38.46,33.82,30.46,28.12,23.06,18.98,18.12.HR-MS(ESI):Calculated for C₂₇H₂₆Cl₂O_6.[M+NH₄]⁺:534.1450.found:534.1448.

Example 16

Compound 5a

The same procedure used in example 2 was repeated except for using 151mg of 2-bromobenzoic acid instead of benzoic acid to give the product as a white solid in 71% yield.¹H NMR(400MHz,DMSO-d₆)δ7.76(dd,J＝5.8,3.4Hz,1H),7.71(dd,J＝6.0,3.6Hz,1H),7.51(dd,J＝5.9,3.4Hz,2H),6.27(s,1H),5.95(s,1H),5.06(s,1H),4.84(dt,J＝4.4,2.2Hz,1H),3.55(d,J＝8.2Hz,1H),3.31(s,1H),2.95–2.91(m,1H),2.90–2.87(m,1H),2.84(d,J＝8.3Hz,1H),2.73(d,J＝16.4Hz,1H),1.83(ddd,J＝10.5,7.5,3.7Hz,1H),1.66(dd,J＝7.8,2.9Hz,1H),1.40(s,1H),1.38–1.35(m,1H),1.25(d,J＝10.6Hz,1H),1.10(d,J＝3.2Hz,1H),1.08(d,J＝6.6Hz,1H),0.82(s,3H),0.72(d,J＝3.0Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.27,198.28,173.94,164.21,146.04,134.11,133.81,131.18,130.77,128.02,123.20,120.21,74.93,72.54,58.95,56.31,46.52,45.28,43.17,39.75,38.51,33.82,30.49,28.14,23.14,18.98,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇BrO_6.[M+NH₄]⁺:544.1335.found:544.1334.

Example 17

Compound 5b

The same procedure used in example 2 was repeated except for using 151mg of 3-bromobenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 65%.¹H NMR(400MHz,DMSO-d₆)δ7.94(t,J＝1.8Hz,1H),7.92–7.89(m,1H),7.86(dt,J＝7.9,1.4Hz,1H),7.51(t,J＝7.9Hz,1H),6.29(s,1H),5.93(s,1H),5.05(d,J＝1.2Hz,1H),4.88(dd,J＝4.0,1.7Hz,1H),3.51(d,J＝8.1Hz,1H),3.31(s,1H),2.91(dd,J＝4.2,1.8Hz,1H),2.90–2.87(m,1H),2.83(t,J＝8.2Hz,1H),2.72(d,J＝16.4Hz,1H),1.84–1.78(m,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.41(dd,J＝7.0,3.0Hz,1H),1.37(s,1H),1.25(d,J＝11.1Hz,1H),1.11(d,J＝3.4Hz,1H),1.08(t,J＝3.9Hz,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.25,197.97,173.95,164.12,163.60,145.82,138.48,129.50,127.21,123.23,116.45,114.64,75.08,72.27,59.01,56.68,56.23,46.42,45.18,43.06,39.80,38.08,33.81,30.51,28.12,23.19,19.02,18.16.HR-MS(ESI):Calculated for C₂₇H₂₇BrO_6.[M+H]⁺:527.1069.found:527.1063.

Example 18

Compound 5c

The same procedure used in example 2 was repeated except for using 151mg of 4-bromobenzoic acid instead of benzoic acid to give the product as a white solid in 67% yield.¹H NMR(400MHz,DMSO-d₆)δ7.77(d,J＝3.1Hz,4H),6.28(s,1H),5.93(s,1H),5.03(s,1H),4.86–4.82(m,1H),3.51(d,J＝8.1Hz,1H),2.91(t,J＝12.4Hz,2H),2.83(t,J＝8.3Hz,1H),2.72(d,J＝16.4Hz,1H),1.85–1.77(m,1H),1.69(d,J＝9.4Hz,1H),1.66–1.59(m,1H),1.43–1.36(m,2H),1.25(d,J＝10.0Hz,1H),1.08(dd,J＝13.7,9.6Hz,2H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.31,198.40,173.91,163.72,146.04,132.09(*2),131.22(*2),128.18,127.56,123.09,74.29,72.48,58.93,56.38,46.55,45.33,43.20,38.52,33.83,30.51,28.16,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇BrO_6.[M+H]⁺:527.1069.found:527.1064.

Example 19

Compound 6a

The same procedures used in example 2 were repeated except for using 105mg of 2-fluorobenzoic acid instead of the benzoic acid to give the product as a white solid in a yield of 85%.¹H NMR(400MHz,DMSO-d₆)δ7.83(td,J＝7.6,1.7Hz,1H),7.72(dddd,J＝7.9,6.9,5.0,1.9Hz,1H),7.38–7.33(m,2H),6.27(d,J＝1.3Hz,1H),5.92(d,J＝1.1Hz,1H),5.05(d,J＝1.1Hz,1H),4.83–4.79(m,1H),3.53(d,J＝8.2Hz,1H),2.94–2.88(m,2H),2.84(t,J＝8.3Hz,1H),2.72(d,J＝16.4Hz,1H),2.51(d,J＝2.0Hz,1H),1.84(ddd,J＝10.5,7.5,3.8Hz,1H),1.70–1.66(m,1H),1.60(dt,J＝14.2,3.6Hz,1H),1.41(d,J＝12.2Hz,1H),1.37(s,1H),1.15–1.10(m,1H),1.09–1.05(m,1H),0.82(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.30,198.29,173.90,162.38,162.13,159.81,146.02,136.10,131.86,124.85,122.97,117.32,116.87,74.40,72.52,58.83,56.32,46.54,45.33,43.21,38.50,33.84,30.49,28.16,23.10,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇FO_6.[M+NH₄]⁺:484.2135.found:484.2131.

Example 20

Compound 6b

The same procedures used in example 2 were repeated except for using 105mg of 3-fluorobenzoic acid instead of the benzoic acid to give a white solid product in a yield of 82%.¹H NMR(400MHz,DMSO-d₆)δ7.72(dt,J＝7.2,1.6Hz,1H),7.62(dd,J＝5.1,2.7Hz,1H),7.60(q,J＝2.0Hz,1H),7.59–7.56(m,1H),6.29(d,J＝1.2Hz,1H),5.93(d,J＝1.1Hz,1H),5.04(d,J＝1.2Hz,1H),4.92–4.88(m,1H),3.52(d,J＝8.2Hz,1H),2.92(dd,J＝4.1,1.8Hz,1H),2.90–2.87(m,1H),2.86–2.80(m,1H),2.72(d,J＝16.5Hz,1H),2.51(d,J＝1.9Hz,1H),1.81(ddd,J＝14.6,7.5,4.1Hz,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.65–1.56(m,1H),1.43–1.39(m,1H),1.37(d,J＝3.2Hz,1H),1.12(q,J＝4.6,3.8Hz,1H),1.09–1.04(m,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.30,198.37,173.94,163.30,163.11,160.67,146.02,131.28,130.65,125.55,123.08,121.20,115.96,74.48,72.49,58.96,56.38,46.54,45.35,43.21,38.51,33.83,30.51,28.16,23.16,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇FO_6.[M+NH₄]⁺:484.2135.found:484.2135.

Example 21

Compound 6c

The same procedures used in example 2 were repeated except for using 105mg of 4-fluorobenzoic acid instead of the benzoic acid to give a white solid product in a yield of 76%.¹H NMR(400MHz,DMSO-d₆)δ7.96–7.92(m,2H),7.39–7.35(m,2H),6.28(s,1H),5.92(s,1H),5.02(d,J＝1.1Hz,1H),4.88–4.84(m,1H),3.50(d,J＝8.2Hz,1H),2.93–2.87(m,2H),2.83(t,J＝8.3Hz,1H),2.72(d,J＝16.4Hz,1H),2.51(s,1H),1.82(ddd,J＝14.5,7.6,4.1Hz,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.59(d,J＝13.8Hz,1H),1.40(t,J＝3.0Hz,1H),1.37(s,1H),1.11(d,J＝3.5Hz,1H),1.09–1.04(m,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.34,198.44,173.92,166.72,163.40,146.08,132.36,132.26,132.10,124.95,123.04,116.24,116.02,115.68,115.46,74.16,72.51,58.92,56.38,46.54,45.34,43.21,38.55,33.83,30.51,28.16,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇FO_6.[M+NH₄]⁺:484.2135.found:484.2131.

Example 22

Compound 6d

The same procedure used in example 2 was repeated except for using 133mg of 2,3, 4-trifluorobenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 76%.¹H NMR(400MHz,DMSO-d₆)δ7.72(tdd,J＝8.2,5.8,2.3Hz,1H),7.51–7.44(m,1H),6.27(s,1H),5.93(s,1H),5.07(s,1H),4.83–4.80(m,1H),3.53(d,J＝8.1Hz,1H),3.33(s,1H),2.93–2.89(m,1H),2.87(d,J＝3.9Hz,1H),2.84(t,J＝8.2Hz,1H),2.72(d,J＝16.4Hz,1H),1.82(ddd,J＝14.5,7.5,4.1Hz,1H),1.65(s,1H),1.60(td,J＝10.4,5.2Hz,1H),1.41(dd,J＝5.8,2.4Hz,1H),1.37(s,1H),1.15–1.10(m,1H),1.09–1.04(m,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.25,198.16,173.90,160.65,145.90,126.91,126.87,126.82,126.78,123.09,113.25,113.11,74.80,72.43,58.84,56.33,46.52,45.30,43.18,38.46,33.83,30.49,28.13,23.10,18.93,18.14.HR-MS(ESI):Calculated for C₂₇H₂₅F₃O_6.[M+NH₄]⁺:520.1947.found:520.1905.

Example 23

Compound 6e

The same procedure used in example 2 was repeated except for using 160mg of 2,3,4,5, 6-pentafluorobenzoic acid in place of benzoic acid to give a white solid as a product in 54% yield.¹H NMR(400MHz,DMSO-d₆)δ6.27(s,1H),5.98–5.93(m,1H),5.14(s,1H),4.75(dt,J＝5.8,2.9Hz,1H),3.54(d,J＝8.1Hz,1H),2.92–2.83(m,3H),2.72(d,J＝16.5Hz,1H),1.86–1.80(m,1H),1.67(dd,J＝10.6,7.5Hz,1H),1.63–1.54(m,1H),1.43–1.39(m,1H),1.37(s,1H),1.10(d,J＝3.4Hz,1H),1.09–1.04(m,1H),0.82(s,3H),0.72(d,J＝2.8Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.03,197.89,173.82,145.72,123.20,75.73,72.32,58.84,56.31,46.50,45.28,43.19,38.48,33.83,30.47,28.12,23.04,18.92,18.14.HR-MS(ESI):Calculated for C₂₇H₂₃F₅O_6.[M+NH₄]⁺:556.1759.found:556.1754.

Example 24

Compound 7a

The same procedures used in example 2 were repeated except for using 143mg of 2-trifluoromethylbenzoic acid instead of benzoic acid to obtain a white solid product with a yield of 48%.¹H NMR(400MHz,DMSO-d₆)δ7.89(dd,J＝6.5,3.3Hz,1H),7.82(s,1H),7.82(s,1H),7.80(d,J＝3.5Hz,1H),6.27(s,1H),5.96(s,1H),5.06(d,J＝1.1Hz,1H),4.74(q,J＝1.8Hz,1H),3.53(d,J＝8.2Hz,1H),3.33(s,1H),2.92(d,J＝5.0Hz,1H),2.88(d,J＝8.3Hz,1H),2.86–2.82(m,1H),2.74(d,J＝16.4Hz,1H),1.84–1.78(m,1H),1.67(dd,J＝10.4,7.4Hz,1H),1.60(dt,J＝14.3,3.5Hz,1H),1.42–1.39(m,1H),1.37(s,1H),1.10(dd,J＝6.7,2.5Hz,1H),1.07(d,J＝9.3Hz,1H),0.81(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.18,173.90,164.72,145.87,133.00,132.51,130.28,126.85,126.80,123.08,75.21,72.36,58.95,56.22,46.55,45.25,43.16,38.30,33.81,30.48,28.13,23.10,18.95,18.14.HR-MS(ESI):Calculated for C₂₈H₂₇FO_6.[M+NH₄]⁺:534.2103.found:534.2096.

Example 25

Compound 7b

The same operation as in example 2 was carried out using 143mg of 3-trifluoromethylbenzoic acid instead of benzoic acid to give the product as a white solid with a yield of 52%.¹H NMR(400MHz,DMSO-d₆)δ8.15(d,J＝7.9Hz,1H),8.10–8.06(m,2H),7.80(t,J＝8.1Hz,1H),6.30(s,1H),5.93(s,1H),5.10(s,1H),4.92–4.87(m,1H),3.54(d,J＝8.1Hz,1H),2.95–2.88(m,2H),2.87–2.81(m,1H),2.73(d,J＝16.4Hz,1H),1.81(ddd,J＝14.5,7.5,4.1Hz,1H),1.69(dd,J＝10.6,7.6Hz,1H),1.66–1.57(m,1H),1.42(d,J＝12.3Hz,1H),1.36(d,J＝12.5Hz,1H),1.25(d,J＝11.4Hz,1H),1.16–1.11(m,1H),1.10–1.05(m,1H),0.83(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.27,198.35,173.93,163.19,146.05,133.26,130.44,129.59,125.54,125.50,123.08,74.60,72.45,58.99,56.43,46.57,45.34,43.21,38.54,33.83,30.50,28.16,23.18,18.94,18.16.HR-MS(ESI):Calculated for C₂₈H₂₇FO_6.[M+NH₄]⁺:534.2103.found:534.2097.

Example 26

Compound 7c

The same operation as in example 2 was carried out using 143mg of 4-trifluoromethylbenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 67%.¹H NMR(400MHz,DMSO-d₆)δ8.07(d,J＝8.2Hz,2H),7.92(d,J＝8.3Hz,2H),6.30(s,1H),5.94(s,1H),5.10(s,1H),4.90–4.86(m,1H),3.55(d,J＝8.1Hz,1H),3.33(s,1H),2.95–2.89(m,2H),2.85(d,J＝8.2Hz,1H),2.76(s,1H),1.85–1.79(m,1H),1.72–1.67(m,1H),1.63–1.59(m,1H),1.39(d,J＝12.6Hz,2H),1.12(dt,J＝8.4,5.2Hz,2H),0.83(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.27,173.93,163.31,145.97,132.15,130.18,125.91,125.88,125.84,123.13,74.57,72.48,58.98,56.42,46.56,45.34,43.21,39.74,38.52,33.81,30.47,28.15,23.14,18.92,18.14.HR-MS(ESI):Calculated for C₂₈H₂₇FO_6.[M+NH₄]⁺:534.2103.found:534.2100.

Example 27

Compound 8a

The same procedure used in example 2 was repeated except for using 126mg of 2-nitrobenzoic acid in place of benzoic acid to give the product as a white solid in 85% yield.¹H NMR(400MHz,DMSO-d₆)δ8.10–8.07(m,1H),7.87(d,J＝3.7Hz,1H),7.86–7.85(m,1H),7.84(s,1H),6.25(s,1H),5.95(s,1H),5.06(s,1H),4.74–4.71(m,1H),3.54(d,J＝8.3Hz,1H),3.30(s,1H),2.93–2.86(m,2H),2.85–2.82(m,1H),2.73(d,J＝16.4Hz,1H),1.83–1.77(m,1H),1.65(d,J＝3.0Hz,1H),1.40(d,J＝4.3Hz,1H),1.38–1.36(m,1H),1.23(s,1H),1.10(d,J＝3.6Hz,1H),1.07(d,J＝4.5Hz,1H),0.81(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.23,197.93,173.96,163.53,147.28,145.68,133.92,133.31,129.93,125.11,124.26,123.32,75.25,72.34,58.99,56.26,46.45,45.21,43.11,39.75,38.19,33.80,30.48,28.10,23.12,18.97,18.13.HR-MS(ESI):Calculated for C₂₇H₂₇NO_8.[M+NH₄]⁺:511.2080.found:511.2077.

Example 28

Compound 8b

The procedure of example 2 was repeated except for using 126mg of 3-nitrobenzoic acid in place of benzoic acid to obtain a white solid product,the yield thereof was found to be 79%.¹H NMR(400MHz,DMSO-d₆)δ8.51(d,J＝8.2Hz,2H),8.29–8.25(m,1H),7.85(t,J＝7.9Hz,1H),6.31(s,1H),5.95(s,1H),5.12(s,1H),4.91–4.88(m,1H),3.56(d,J＝8.1Hz,1H),3.33(s,1H),2.92(q,J＝1.9Hz,1H),2.89(d,J＝2.6Hz,1H),2.88–2.83(m,1H),2.74(d,J＝16.5Hz,1H),1.84–1.78(m,1H),1.69(dd,J＝10.5,7.5Hz,1H),1.60(dt,J＝12.3,2.7Hz,1H),1.43–1.40(m,1H),1.38(s,1H),1.12(d,J＝3.9Hz,1H),1.09(t,J＝4.8Hz,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.30,198.32,173.97,162.74,147.88,145.93,135.33,130.84,129.97,128.30,123.71,123.25,74.79,72.44,58.99,56.43,46.57,45.31,43.19,39.74,38.52,33.82,30.49,28.14,23.16,18.93,18.14.HR-MS(ESI):Calculated for C₂₇H₂₇NO_8.[M+NH₄]⁺:511.2080.found:511.2086.

Example 29

Compound 8c

The same procedure used in example 2 was repeated except for using 126mg of 4-nitrobenzoic acid in place of benzoic acid to give the product as a white solid in 71% yield.¹H NMR(400MHz,DMSO-d₆)δ8.35(d,J＝2.1Hz,1H),8.33(d,J＝2.4Hz,1H),8.11(d,J＝2.3Hz,1H),8.09(q,J＝2.2Hz,1H),6.30(s,1H),5.94(s,1H),5.09(s,1H),4.91–4.87(m,1H),3.56(d,J＝8.2Hz,1H),3.33(s,1H),2.92(q,J＝1.9Hz,1H),2.89(s,1H),2.84(d,J＝8.2Hz,1H),2.74(d,J＝16.5Hz,1H),1.85–1.79(m,1H),1.69(dd,J＝10.5,7.4Hz,1H),1.60(dt,J＝14.2,3.7Hz,1H),1.41(s,1H),1.39–1.36(m,1H),1.12(d,J＝4.0Hz,1H),1.09(d,J＝4.6Hz,1H),0.83(s,3H),0.72(d,J＝3.8Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.29,198.30,173.96,162.98,150.52,145.89,133.77,130.82(*2),123.95(*2),123.26,74.83,72.46,59.00,56.41,46.56,45.32,43.20,39.73,38.48,33.82,30.48,28.14,23.14,18.93,18.14.HR-MS(ESI):Calculated for C₂₇H₂₇NO_8.[M+NH₄]⁺:511.2080.found:511.2079.

Example 30

Compound 8d

The same procedure used in example 2 was repeated except for using 160mg of 2, 4-dinitrobenzoic acid in place of benzoic acid to give a white solid product in a yield of 64%.¹H NMR(400MHz,DMSO-d₆)δ8.81(dd,J＝4.3,2.2Hz,1H),8.64(ddd,J＝8.0,5.7,2.2Hz,1H),8.14(d,J＝8.5Hz,1H),6.25(d,J＝1.2Hz,1H),5.96(d,J＝1.2Hz,1H),5.11(d,J＝1.2Hz,1H),4.75(dd,J＝3.9,1.7Hz,1H),3.62–3.56(m,1H),2.89(d,J＝8.7Hz,1H),2.87–2.84(m,1H),2.74(d,J＝16.5Hz,1H),1.84–1.73(m,2H),1.64(dt,J＝10.4,6.5Hz,2H),1.39(s,1H),1.37(s,1H),1.23(s,1H),1.12–1.09(m,1H),1.08–1.05(m,1H),0.81(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.09,197.76,173.90,162.36,149.20,147.10,145.60,131.62,130.11,128.50,123.43,119.90,75.94,72.24,59.04,56.30,46.45,45.22,43.12,39.78,38.18,33.82,30.50,28.11,23.12,18.99,18.15.HR-MS(ESI):Calculated for C₂₇H₂₆N₂O_10.[M+NH₄]⁺:556.1931.found:556.1926.

Example 31

Compound 8e

The same procedure used in example 2 was repeated except for using 160mg of 3, 4-dinitrobenzoic acid instead of benzoic acid to give a white solid product in 56% yield.¹H NMR(400MHz,DMSO-d₆)δ8.54(d,J＝3.0Hz,1H),8.35(d,J＝4.1Hz,2H),6.30(s,1H),5.93(d,J＝5.6Hz,1H),5.10(s,1H),4.99–4.95(m,1H),3.56(d,J＝8.1Hz,1H),2.92(d,J＝3.8Hz,1H),2.89(dd,J＝5.7,2.3Hz,1H),2.86–2.82(m,1H),2.73(d,J＝16.5Hz,1H),1.82–1.76(m,1H),1.72–1.67(m,1H),1.42–1.40(m,1H),1.38(s,1H),1.25(d,J＝10.7Hz,1H),1.11(d,J＝4.3Hz,1H),1.09(t,J＝4.1Hz,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.19,173.96,145.83,141.75,135.24,133.51,126.34,126.22,123.31,75.52,72.30,59.12,56.40,46.61,45.36,43.22,38.40,33.83,30.51,28.15,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₆N₂O_10.[M+H]⁺:539.1666.found:539.1652.

Example 32

Compound 8f

The same procedure used in example 2 was repeated except for using 160mg of 3, 5-dinitrobenzoic acid in place of benzoic acid to give a white solid product in 74% yield.¹H NMR(400MHz,DMSO-d₆)δ9.06–9.02(m,1H),8.78(d,J＝2.1Hz,2H),6.33(s,1H),5.96(s,1H),5.18(s,1H),4.96–4.92(m,1H),3.58(d,J＝8.1Hz,1H),3.34(s,1H),2.91(d,J＝3.9Hz,1H),2.90–2.84(m,2H),2.75(d,J＝16.5Hz,1H),1.81(ddd,J＝11.0,7.6,3.8Hz,1H),1.72–1.68(m,1H),1.39(d,J＝12.7Hz,2H),1.25(d,J＝10.2Hz,1H),1.12(s,1H),1.10–1.08(m,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.18,198.18,173.92,161.36,148.26(*2),145.90,131.51,128.98(*2),123.36,122.92,75.48,72.29,59.06,56.48,46.60,45.30,43.19,39.76,38.51,33.84,30.51,28.14,23.17,18.94,18.16.HR-MS(ESI):Calculated for C₂₇H₂₆N₂O_10.[M+NH₄]⁺:556.1931.found:556.1921.

Example 33

Compound 8j

The same procedure used in example 2 was repeated except for using 149mg of 2-nitro-5-methoxybenzoic acid in place of benzoic acid to give a white solid product in a yield of 69%.¹H NMR(400MHz,DMSO-d₆)δ8.17–8.14(m,1H),7.30(d,J＝3.3Hz,1H),7.29–7.27(m,1H),6.23(s,1H),5.94(d,J＝1.1Hz,1H),5.07(d,J＝1.2Hz,1H),4.74(dd,J＝3.9,1.7Hz,1H),3.94(d,J＝8.8Hz,1H),3.92(s,3H),3.58(d,J＝8.4Hz,1H),3.30(s,1H),2.90–2.86(m,1H),2.86–2.82(m,1H),2.74(d,J＝16.4Hz,1H),1.81–1.75(m,1H),1.67–1.61(m,2H),1.40(d,J＝3.8Hz,1H),1.37(s,1H),1.09(d,J＝4.2Hz,1H),1.06(d,J＝4.4Hz,1H),0.81(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.15,197.86,173.94,145.68,127.85,127.75,123.35,119.55,117.24,116.98,75.54,72.23,59.05,56.25,46.44,45.2,43.10,39.79,38.12,33.81,30.51,28.12,23.15,19.00,18.15.HR-MS(ESI):Calculated for C₂₈H₂₉NO_9.[M+H]⁺:524.1921.found:524.1906.

Example 34

Compound 8h

The same operation as in example 2 was carried out using 137mg of 2-nitro-5-methylbenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 78%.¹H NMR(400MHz,DMSO-d₆)δ8.04–7.99(m,1H),7.62(d,J＝7.0Hz,2H),6.24(s,1H),5.95(s,1H),5.05(s,1H),4.73(d,J＝3.9Hz,1H),3.53(d,J＝8.7Hz,1H),3.30(s,1H),2.89(d,J＝7.7Hz,1H),2.85(d,J＝7.7Hz,1H),2.76–2.70(m,1H),2.45(s,3H),1.82–1.76(m,1H),1.64(dt,J＝11.3,5.7Hz,2H),1.40(s,1H),1.11–1.06(m,2H),0.81(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.18,197.96,173.91,164.07,145.75,145.54,144.52,132.99,129.75,126.01,124.40,123.25,75.15,72.29,59.01,56.23,46.48,45.24,43.12,38.15,33.82,30.50,28.13,23.15,20.73,18.99,18.15.HR-MS(ESI):Calculated for C₂₈H₂₉NO_8.[M+NH₄]⁺:525.2237.found:525.2233.

Example 35

Compound 8i

The same procedure used in example 2 was repeated except for using 152mg of 2-nitro-5-chlorobenzoic acid in place of the benzoic acid to give a white solid product in a yield of 70%.¹H NMR(400MHz,DMSO-d₆)δ8.15(d,J＝8.7Hz,1H),7.98(d,J＝2.3Hz,1H),7.92(dd,J＝8.7,2.3Hz,1H),6.25(s,1H),5.96(s,1H),5.07(s,1H),4.81–4.77(m,1H),3.56(d,J＝8.1Hz,1H),3.30(s,1H),2.93–2.81(m,3H),2.74(d,J＝16.4Hz,1H),1.78(ddd,J＝14.6,7.6,4.0Hz,1H),1.65(dd,J＝10.6,7.6Hz,1H),1.43–1.35(m,2H),1.31–1.21(m,1H),1.12(dd,J＝13.6,4.1Hz,1H),1.05(dd,J＝13.8,4.4Hz,1H),0.81(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.15,197.85,173.93,162.59,145.69,145.23,138.88,132.71,129.42,126.40,123.34,75.58,72.21,59.05,56.25,46.44,45.19,43.07,39.79,38.10,33.81,30.50,28.11,23.16,18.99,18.15.HR-MS(ESI):Calculated for C₂₇H₂₆ClNO_8.[M+NH₄]⁺:545.1691.found:545.1683.

Example 36

Compound 8g

The same procedures used in example 2 were repeated except for using 140mg of 2-nitro-5-fluorobenzoic acid in place of the benzoic acid to give a white solid product in a yield of 66%.¹H NMR(400MHz,DMSO-d₆)δ8.24(dd,J＝9.0,4.6Hz,1H),7.81(dd,J＝8.2,2.9Hz,1H),7.70(ddd,J＝9.0,7.8,2.8Hz,1H),6.25(d,J＝1.3Hz,1H),5.95(d,J＝1.2Hz,1H),5.07(d,J＝1.2Hz,1H),4.78(dt,J＝4.1,2.2Hz,1H),3.59–3.53(m,1H),3.30(s,1H),2.92–2.88(m,1H),2.86–2.83(m,1H),2.74(d,J＝16.5Hz,1H),1.77(ddd,J＝13.4,6.7,2.6Hz,1H),1.68–1.60(m,2H),1.43–1.40(m,1H),1.37(d,J＝2.6Hz,1H),1.25(d,J＝11.1Hz,1H),1.10(d,J＝4.0Hz,1H),1.07(d,J＝4.2Hz,1H),0.81(s,3H),0.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.29,198.40,173.95,163.12,146.04,136.74,131.55,131.18,130.62,128.39,123.12,121.91,74.51,72.48,58.99,56.40,46.57,45.36,43.22,38.53,33.84,30.52,28.17,23.19,18.96,18.17.HR-MS(ESI):Calculated for C₂₇H₂₆FNO_8.[M+NH₄]⁺:529.1986.found:529.1934.

Example 37

Compound 8k

By 140mg of 3-nitro-4-fluorobenzoic acid was used in place of benzoic acid, and the procedure was as in example 2 to give a white solid product in 54% yield.¹H NMR(400MHz,DMSO-d₆)δ8.48(dd,J＝7.1,2.3Hz,1H),8.25(ddd,J＝8.8,4.2,2.3Hz,1H),7.75(dd,J＝10.9,8.8Hz,1H),6.30(s,1H),5.97–5.91(m,1H),5.08(d,J＝1.2Hz,1H),4.94–4.89(m,1H),3.54(d,J＝8.1Hz,1H),2.96–2.87(m,2H),2.87–2.81(m,1H),2.73(d,J＝16.5Hz,1H),1.80(ddd,J＝14.5,7.5,4.1Hz,1H),1.69(dd,J＝10.5,7.4Hz,1H),1.65–1.56(m,1H),1.41(d,J＝8.3Hz,1H),1.39–1.35(m,1H),1.15–1.10(m,1H),1.10–1.04(m,1H),0.83(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.22,198.26,173.92,162.04,145.93,137.14,137.06,137.01,136.90,127.41,125.54,123.21,119.61,119.39,74.95,72.38,59.02,56.41,46.57,45.35,43.22,38.50,33.83,30.51,28.15,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₆FNO_8.[M+NH₄]⁺:529.1986.found:529.1980.

Example 38

Compound 9a

The same procedure used in example 2 was repeated except for using 187mg of 2-iodobenzoic acid in place of the benzoic acid to give the product as a white solid in a yield of 76%.¹H NMR(400MHz,DMSO-d₆)δ7.79(d,J＝7.8Hz,1H),7.39(dd,J＝7.8,1.8Hz,1H),7.28(t,J＝7.6Hz,1H),7.09–7.05(m,1H),6.04(s,1H),5.72(s,1H),4.81(s,1H),4.63–4.60(m,1H),3.31(d,J＝8.2Hz,1H),3.07(s,1H),2.70–2.64(m,2H),2.61(d,J＝8.2Hz,1H),2.50(d,J＝16.4Hz,1H),1.58(ddd,J＝14.7,7.6,4.1Hz,1H),1.45–1.41(m,1H),1.17(d,J＝11.4Hz,1H),1.15–1.11(m,1H),1.01(d,J＝10.3Hz,1H),0.91–0.86(m,1H),0.85–0.80(m,1H),0.58(s,3H),0.49(d,J＝3.3Hz,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.31,198.29,173.98,164.96,145.98,140.81,134.10,133.53,130.43,128.40,123.33,94.27,74.82,72.56,58.97,56.31,46.49,45.26,43.16,39.74,38.51,33.82,30.49,28.13,23.15,18.98,18.14.HR-MS(ESI):Calculated for C₂₇H₂₇IO_6.[M+NH₄]⁺:592.1196.found:592.1192.

Example 39

Compound 9b

The same operation as in example 2 was carried out using 187mg of 3-iodobenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 82%.¹H NMR(400MHz,DMSO-d₆)δ8.11(t,J＝1.7Hz,1H),8.06(dt,J＝7.9,1.3Hz,1H),7.86(dq,J＝8.0,2.2,1.8Hz,1H),7.34(t,J＝7.9Hz,1H),6.29(s,1H),5.93(s,1H),5.04(d,J＝1.2Hz,1H),4.88–4.84(m,1H),3.50(d,J＝8.2Hz,1H),3.31(s,1H),2.94–2.86(m,2H),2.83(t,J＝8.2Hz,1H),2.72(d,J＝16.4Hz,1H),1.81(ddd,J＝14.4,7.5,4.0Hz,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.40(s,1H),1.37(s,1H),1.25(d,J＝10.6Hz,1H),1.15–1.10(m,1H),1.09–1.04(m,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.31,198.41,173.94,163.05,146.04,142.49,137.31,131.01,130.43,128.63,123.11,94.90,74.40,72.47,58.96,56.39,46.56,45.33,43.20,38.52,33.83,30.51,28.16,23.18,18.95,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇IO_6.[M+NH₄]⁺:592.1196.found:592.1191.

Example 40

Compound 9c

The same operation as in example 2 was carried out using 187mg of 4-iodobenzoic acid in place of benzoic acid to give the product as a white solid in a yield of 78%.¹H NMR(400MHz,DMSO-d₆)δ7.95–7.92(m,2H),7.60(dd,J＝8.6,2.0Hz,2H),6.28(s,1H),5.92(s,1H),5.03(s,1H),4.83(dd,J＝4.0,1.7Hz,1H),3.50(d,J＝8.2Hz,1H),3.31(s,1H),2.94–2.90(m,1H),2.88(t,J＝3.4Hz,1H),2.83(t,J＝8.2Hz,1H),2.72(d,J＝16.4Hz,1H),1.82(ddd,J＝10.8,7.6,4.0Hz,1H),1.68(dd,J＝10.5,7.5Hz,1H),1.42–1.39(m,1H),1.35(d,J＝11.4Hz,1H),1.23(s,1H),1.12(q,J＝4.8,3.6Hz,1H),1.06(dd,J＝13.7,4.5Hz,1H),0.82(s,3H),0.71(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.29,198.40,173.90,164.05,146.04,137.94(*2),130.86(*2),127.82,123.07,102.77,74.23,72.49,58.94,56.38,46.56,45.35,43.21,38.53,33.83,30.51,28.16,23.15,18.94,18.15.HR-MS(ESI):Calculated for C₂₇H₂₇IO_6.[M+NH₄]⁺:592.1196.found:592.1193.

EXAMPLE 41

Compound 10a

The same procedures used in example 2 were repeated except for using 162mg of 3-bromomethylbenzoic acid instead of benzoic acid to give a product as a white solid in a yield of 37%.¹H NMR(400MHz,DMSO-d₆)δ7.96(q,J＝2.4,1.8Hz,1H),7.82(dt,J＝7.7,1.5Hz,1H),7.78(s,1H),7.56(t,J＝7.8Hz,1H),6.28(s,1H),5.92(s,1H),5.03(s,1H),4.87(dd,J＝4.1,1.7Hz,1H),3.52(d,J＝8.1Hz,1H),3.32(s,1H),2.91(ddd,J＝16.2,5.0,2.4Hz,2H),2.84(t,J＝8.2Hz,1H),2.72(d,J＝16.5Hz,1H),1.83(ddd,J＝14.6,7.5,4.1Hz,1H),1.72–1.68(m,1H),1.66–1.61(m,1H),1.40(s,1H),1.37(s,1H),1.25(d,J＝9.6Hz,1H),1.12–1.09(m,1H),0.83(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.35,198.48,173.95,164.06,146.07,138.68,134.39,129.45,129.17,128.80,123.08,74.31,72.54,58.99,56.40,46.61,45.39,45.13,43.26,38.54,33.85,30.52,28.18,23.18,18.95,18.16.HR-MS(ESI):Calculated for C₂₈H₂₉BrO_6.[M+NH₄]⁺:541.1226.found:541.1210.

Example 42

Compound 10b

The same operation as in example 2 was carried out using 162mg of 4-bromomethylbenzoic acid instead of benzoic acid to give the product as a white solid in a yield of 27%.¹H NMR(400MHz,DMSO-d₆)δ7.78(d,J＝8.2Hz,2H),7.49(d,J＝8.3Hz,2H),6.28(s,1H),5.92(s,1H),5.03(s,1H),4.87(dd,J＝4.1,1.7Hz,1H),3.52(d,J＝8.1Hz,1H),3.32(s,1H),2.91(ddd,J＝16.2,5.0,2.4Hz,2H),2.84(t,J＝8.2Hz,1H),2.72(d,J＝16.5Hz,1H),1.83(ddd,J＝14.6,7.5,4.1Hz,1H),1.72–1.68(m,1H),1.66–1.61(m,1H),1.40(s,1H),1.37(s,1H),1.25(d,J＝9.6Hz,1H),1.12–1.09(m,1H),0.83(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.35,198.48,173.95,164.06,146.07,129.59(*2),128.58(*2),112.89,77.04,72.04,66.09,62.79,61.99,59.14,52.91,46.61,45.39,45.13,43.26,38.54,33.85,30.52,28.18,23.18,18.95,18.16.HR-MS(ESI):Calculated for C₂₈H₂₉BrO_6.[M+NH₄]⁺:541.1226.found:541.1230.

Example 43

Compound 11a

The same procedures used in example 2 were repeated except for using 116mg of 2-methyl-3-fluorobenzoic acid in place of the benzoic acid to give a white solid product in a yield of 37%.¹H NMR(400MHz,DMSO-d₆)δ7.53(d,J＝7.8Hz,1H),7.44(d,J＝9.1Hz,1H),7.39–7.34(m,1H),6.29(s,1H),5.95(s,1H),5.05(s,1H),4.82–4.79(m,1H),3.54(d,J＝8.1Hz,1H),3.31(s,1H),2.90(dd,J＝10.7,6.8Hz,2H),2.83(d,J＝8.3Hz,1H),2.73(d,J＝16.4Hz,1H),2.41(d,J＝2.4Hz,1H),2.34(d,J＝2.2Hz,3H),1.82(td,J＝7.3,4.0Hz,1H),1.70–1.66(m,1H),1.39(dd,J＝14.8,5.8Hz,2H),1.25(d,J＝11.2Hz,1H),1.11(d,J＝3.1Hz,1H),1.08(d,J＝4.6Hz,1H),0.82(s,3H),0.72(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ203.35,198.46,173.95,164.45,161.87,159.46,146.15,130.30,127.57,126.07,123.10,119.69,74.33,72.55,58.88,56.36,46.51,45.28,43.16,39.74,38.55,33.81,30.48,28.15,23.16,18.95,18.14,11.52.HR-MS(ESI):Calculated for C₂₈H₂₉FO_6.[M+NH₄]⁺:498.2292.found:498.2285.

Example 44: the anti-tumor activity evaluation of the synthesized jiyuan oridonin derivative on four cells of human breast cancer cells MCF-7, human pancreatic cancer cells SW-1990, human gastric cancer cells MGC-803 and human esophageal cancer cells Ec109 is as follows: the contents are all mass% contents, not specifically stated.

Experimental methods

Taking oridonin as a positive control, selecting the five cancer cells to determine the anti-proliferative activity of the synthesized compound for 72 hours: digesting the cells in logarithmic growth phase by pancreatin containing 0.25% EDTA to prepare cell suspension with the concentration of 1-10 x 10^ 4/mL, inoculating the cells into a 96-well plate according to 1000-10000 cells/hole, adding 100 mu L of cell suspension into the fast top wall of each hole, and after the inoculation is finished, placing the 96-well plate in a position with the volume percentage content of 5% CO₂In a 37 ℃ humidity incubator. Discarding supernatant in clean plate after 24h, adding 200 μ L culture solution containing different drug concentrations into each well, arranging three parallel wells according to a certain concentration gradient for each compound, adding 200 μ L blank culture solution into control group, and adding 5 vol% CO₂And culturing for 72 hours in a humidity incubator at 37 ℃. Adding 100 μ L of 30% TCA solution into each well, and fixing at 4 deg.C for 60 min; washing with deionized water for 3-4 times, and air drying; adding 0.4% SRB 100 μ L per well, and dyeing at room temperature for 30 min; washing with 1% acetic acid for 3 times, and air drying; adding 150 μ L of Tris base into each well, shaking with a flat plate shaker for 15min, measuring absorbance density (OD) with a microplate reader, and detecting wavelength of 540 nm. Using the solvent control treated cells as a control group, calculating the inhibition rate of the compound on the cells according to the following formula, and calculating the half inhibition concentration IC according to the middle effect equation₅₀: inhibition (%) - (control group OD average-administration group OD average)/control group OD average 100%

Anti-tumor activity of oridonin derivative

Ori is oridonin.

^aA human breast cancer cell line;^bhuman esophageal cancer cell lines;^ca human pancreatic cancer cell line;^dhuman gastric cancer cell line.

The experimental results show that the compounds have stronger in-vitro anti-tumor activity and are stronger than the positive control oridonin. Wherein the compound 8i not only has stronger in vitro anti-tumor activity, but also has good selectivity and stronger inhibition effect on SW1990 cell line (IC)₅₀0.478 μ M). The compound can be further researched and used for preparing antitumor drugs, is used for clinically treating human esophagus cancer, gastric cancer, pancreatic cancer, colorectal cancer, breast cancer and other diseases, and has potential application value.

Test for stability of drug

According to the guiding principle of stability test of 'Chinese pharmacopoeia' 2020 edition, the stability test is carried out on the 8i with the best activity in the 11, 20-dicarbonyl economic source oridonin derivatives. The following tests are required for the starting drug: testing influence factors; carrying out an accelerated test; the long-term test results are shown in the following table:

test for Effect of Compound 8i (1 test sample)

Accelerated test and Long-term test of Compound 8i (3 test batches)

From the above results, it appears that: the compound 8i has better stability and can be further developed and researched.

Drug solubility test

According to solubility test of 'Chinese pharmacopoeia' 2020 edition, the solubility of the series of compounds is determined by taking physiological saline as a solvent, and the result shows that most of the compounds can reach 15mg/mL, and the experimental requirements of animal experiment intravenous injection administration can be met.

Claims

1.11, 20-dicarbonyl economic oridonin 14-O-benzoic acid esterified derivative, which is characterized by having a structure shown in a general formula I:

the R group is mono-substituted, di-substituted or multi-substituted, and R is selected from hydrogen; a methoxy group; halogen; a nitro group; an amino group; a sulfonic acid group; a linear or branched alkyl group; straight chain alkyl substituted with halogen; a trifluoromethyl group.

2. The 11, 20-dicarbonyl oridonin 14-O-benzoic acid esterified derivative of claim 1, wherein the R group is hydrogen; c1-6 straight chain alkyl; c1-6 straight chain alkyl substituted with halogen; halogen; a nitro group; a methoxy group; a trifluoromethyl group.

3. The 11, 20-dicarbonyl oridonin 14-OH benzoic acid esterified series derivative of claim 2, wherein the R group is hydrogen; a methoxy group; a methyl group; chlorine; bromine; fluorine; a trifluoromethyl group; iodine; a nitro group; a bromomethyl group.

4. The 11, 20-dicarbonyl oridonin 14-OH benzoic acid esterified series derivative of claim 1, selected from the group consisting of:

5. the use of 11, 20-dicarbonyl oridonin 14-O-benzoic acid esterified derivative as defined in claim 1 in the preparation of antitumor medicament, wherein the derivative is used as active component in the preparation of medicament for treating esophageal cancer, gastric cancer, breast cancer or pancreatic cancer.