CN113234089A - Limonin compound, preparation method and application thereof as medicine for treating echinococcosis - Google Patents

Limonin compound, preparation method and application thereof as medicine for treating echinococcosis Download PDF

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CN113234089A
CN113234089A CN202110500091.5A CN202110500091A CN113234089A CN 113234089 A CN113234089 A CN 113234089A CN 202110500091 A CN202110500091 A CN 202110500091A CN 113234089 A CN113234089 A CN 113234089A
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limonin
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echinococcosis
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王海
李曹龙
夏元铮
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China Pharmaceutical University
Nanjing Medical University
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Nanjing Medical University
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Abstract

The limonin compounds shown in general formulas I, II and III can be efficiently prepared at normal temperature and normal pressure by adopting hydrazone forming reaction or alkali catalysis reaction, the preparation process is simple, the limonin compounds have excellent curative effect on echinococcosis treatment, the echinococcosis resistant inhibition rate reaches 95%, damaged liver cells can be repaired at the same time, the liver protection effect can be achieved, and the condition of liver and kidney injury caused by medicine application is avoided.

Description

Limonin compound, preparation method and application thereof as medicine for treating echinococcosis
Technical Field
The invention relates to the field of medicinal chemistry, in particular to a limonin compound, a preparation method and application thereof as a medicament for treating echinococcus.
Background
Echinococcosis, also known as echinococcosis, is a serious zoonosis caused by the parasitism of echinococcus larvae (echinococcosis) in human and animal bodies, is distributed worldwide, and is a high-incidence area in China. Echinococcus granulosus, Echinococcus multilocularis and Echinococcus shikoensis are prevalent in China, and among them, echinococcus granulosus larvae cause echinococcus cysticercosis. CE can cause death of human and livestock and cause direct economic loss of over 30 million yuan per year in animal husbandry in China. The CE epidemic areas in China are mainly distributed in vast pasturing areas and farming and pasturing areas in the north and the northwest, and in recent years, the disease has a tendency of spreading from pasturing areas to farming areas. The current treatment means comprises two aspects of surgical treatment and drug treatment, wherein the surgical treatment is often accompanied with the risk of relapse because of the existence of factors for insufficient removal of the small cysts, and the leakage of cyst fluid during puncture causes the sacs or the head nodes to pollute the abdominal cavity, or secondary hydatid cysts formed by rupture of the cysts in the surgical process. In the aspect of drug treatment, internationally adopted treatment drugs are praziquantel and benzimidazole compounds, albendazole is one of representative drugs, the drug has obvious advantages in the aspect of treating echinococcus granulosus and alveolar echinococcus, the survival time and the survival quality of patients are improved, but clinical studies show that the cure rate of the drugs is only 30%, the symptoms of the drugs are improved to a certain extent, the rate of the patients accounts for 30% -50%, and the problems of large individual difference, unstable curative effect and the like exist. Clinical studies show that the albendazole has serious side effects such as encephalitis syndrome, acute liver injury, hemolytic anemia, anaphylactic shock, renal failure and even death, and the like, and in addition, the albendazole mainly has a pest inhibiting effect without a pest killing effect in the process of treating echinococcus granulosus patients. Albendazole needs to be taken for a long time, which increases the economic burden on patients and even the whole society to some extent.
Figure BDA0003056152600000011
The limonoids have typical chemical structures, namely, the limonoids all contain 4,4, 8-trimethyl-17-furan steroid skeleton or are derived from the steroid skeleton, are extracted and separated from natural plants, the limonoids are plant secondary substances with special structures and functions, and are four-carbon-reduced products of triterpenes, all naturally-produced limonoids mainly existing in rue and meliaceae plants are connected with a furan ring at the C-17 position of a D ring, and oxygen-containing functional groups are connected at C-3, C-4, C-7, C-16 and C-17.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the problems that the medicine for treating echinococcosis in the prior art has no insecticidal effect, needs to be taken for a long time and has great toxic and side effects, the invention provides a limonin compound, and also provides a preparation method of the limonin compound and application of the limonin compound as a medicine for treating echinococcosis.
The technical scheme is as follows: the structural general formula of the limonin compound is as follows:
Figure BDA0003056152600000021
wherein X represents: -O-, -CH2-，Bond
R1Represents: C1-C6 alkyl,
Figure BDA0003056152600000022
Figure BDA0003056152600000023
R2All C1-C4 alkyl groups, -X, -CX3、-OCH3、-NH2、-NO2、
Figure BDA0003056152600000024
-OH。
R3All alkyl groups representing C1-C2 ═ CH2。
The preparation method of the limonin compound with the general formula I comprises the following steps:
Figure BDA0003056152600000025
wherein, when the X substituent in the limonin compound of the general formula I is-O-, Bond-CH respectively2When the compound is IV, V and VI; in the process of preparing the compound V from the compound IV, the reaction reagent is hydroiodic acid; the reaction solvent is acetic acid. In the process of preparing the compound VI from the compound V, cuprous chloride is used as a catalyst, and diiodomethane and zinc powder are stirred in ether for 48 hours at the reaction temperature of 40 ℃.
The preparation method of the limonin compound with the general formula II comprises the following steps:
Figure BDA0003056152600000031
the limonin compound of the general formula II is prepared from the limonin compound of the general formula I, the reaction reagent is hydrazine corresponding to alkyl or acyl, the reaction solvent is corresponding methanol or ethanol, acetic acid is used as a catalyst, and the reaction is carried out for 48 hours at 70 ℃.
The preparation method of the limonin compound with the general formula III comprises the following steps:
Figure BDA0003056152600000032
the limonin compound of the general formula III is prepared from the limonin compound of the general formula I, reaction reagents are corresponding iodoalkane and pyrrolidine, solvents are ethanol and benzene, the reaction temperature is 20 ℃, and the reaction time is 3 hours.
The invention also discloses application of the limonin compound as a medicine for treating echinococcosis.
Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable salt formed by the limonin compound and an acid and a pharmaceutically acceptable carrier, wherein the acid is one or more of hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages:
the limonin compound prepared by the invention is used for treating echinococcosis, can efficiently resist echinococcosis, and has an inhibition rate of 95% which is far higher than that of the existing commonly used albendazole by 77.4%; meanwhile, the liver protection effect is achieved, liver and kidney injury caused by medicines used in the prior art is avoided, damaged liver cells can be repaired, and the echinococcosis treatment effect is excellent.
Drawings
FIG. 1: eosin staining pattern of limonin compound and albendazole positive by incubating echinococcus granulosus metacercaria for 3 days in vitro;
FIG. 2: eosin staining pattern of limonin compound and albendazole positive by incubating echinococcus granulosus metacercaria for 5 days in vitro;
FIG. 3: a histogram of in vitro inhibition results of the limonin compounds and albendazole on echinococcus granulosus;
FIG. 4: the result of H.E staining of mouse liver tissue after the intervention of limonin and albendazole and the infection of echinococcus granulosus metacercaria;
FIG. 5: the limonin and albendazole intervene in the H.E staining result of the mouse kidney tissue after the echinococcus granulosus metacercaria is infected;
FIG. 6: the result of H.E staining of mouse cyst tissue after the intervention of limonin and albendazole and the infection of echinococcus granulosus metacercaria;
FIG. 7 is the results of limonin and albendazole intervention in cysts after echinococcus granulosus metacercaria infection;
FIG. 8 is a histogram of the wet weight of the bursa of each group after 3 weeks of drug treatment.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
Example 1
Preparation of Compound II-1
Taking compound limonin as a starting material, adding 0.4725g of limonin and a first component into a 50ml eggplant-shaped bottle in sequence20ml of alcohol, 0.200ml of glacial acetic acid and 0.2148g of thiophene formhydrazide, and stirring is carried out for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-1 as a white solid, 0.2507g, 42% yield. The molecular formula is: c31H34N2O8S，HR-ESUMS m/z：595.2113[M+H]+，1H NMR(300MHz,DMSO-d6)δ10.93(s,1H),7.93(s,1H),7.85(d,J＝5.0Hz,1H),7.75–7.73(m,1H),7.68(t,J＝1.7Hz,1H),7.20(dd,J＝5.0,3.7Hz,1H),6.51(dd,J＝2.0,0.8Hz,1H),5.47(s,1H),4.84(s,1H),4.42(d,J＝12.9Hz,1H),4.10(d,J＝3.6Hz,1H),4.02(s,1H),2.81(d,J＝16.4Hz,1H),2.62(dd,J＝16.4,3.9Hz,1H),2.42(d,J＝15.3Hz,1H),2.25–2.17(m,1H),1.84(d,J＝43.4Hz,4H),1.22(s,3H),1.18(s,3H),1.04(s,3H)。
Preparation of Compound II-2
The compound limonin is used as a starting material, 0.4041g of limonin, 10ml of methanol, 0.100ml of glacial acetic acid and 0.2144g of p-methoxybenzoyl hydrazine are sequentially added into a 50ml eggplant-shaped bottle, and the mixture is stirred for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography on methanol (120:1) gave II-2 as a white solid, 0.3879g, 73% yield. The molecular formula is: c34H38N2O9，HR-ESUMS m/z：641.2482[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.83(s,1H),7.77(d,J＝8.0Hz,2H),7.74(s,1H),7.68(t,J＝1.7Hz,1H),7.31(d,J＝7.9Hz,2H),6.51(s,1H),5.45(s,1H),4.84(d,J＝13.0Hz,1H),4.43(d,J＝12.8Hz,1H),4.09(s,2H),2.82(d,J＝16.5Hz,1H),2.62(dd,J＝16.4,4.0Hz,1H),2.38(s,3H),2.18(d,J＝14.2Hz,1H),1.77(s,4H),1.23(d,J＝3.6Hz,4H),1.17(s,3H),1.03(s,3H),0.99(s,3H)。
Preparation of Compound II-3
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4730g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2300g of 3-hydroxybenzoyl hydrazine, and stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (80:1) gave II-3 as a white solid, 0.3042g, 50% yield. MoleculeThe formula is as follows: c33H36N2O9，HR-ESUMS m/z：627.2319[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.81(s,1H),9.74(s,1H),7.74(d,J＝1.4Hz,1H),7.68(t,J＝1.7Hz,1H),7.29(d,J＝5.5Hz,2H),7.22(s,1H),6.97–6.92(m,1H),6.55–6.49(m,1H),5.45(s,1H),4.84(d,J＝13.0Hz,1H),4.43(d,J＝13.0Hz,1H),4.12–4.09(m,2H),3.18(s,1H),3.16(s,1H),2.82(d,J＝16.4Hz,1H),2.62(dd,J＝16.4,3.9Hz,1H),2.40(d,J＝14.6Hz,1H),2.18(d,J＝14.5Hz,1H),1.77(s,4H),1.20(d,J＝14.8Hz,6H),1.01(d,J＝13.7Hz,6H)。
Preparation of Compound II-4
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4729g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2304g of 4-hydroxybenzoyl hydrazine, and stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-4 as a white solid, 0.2193g, 36% yield. The molecular formula is: c33H36N2O9，HR-ESUMS m/z：627.2322[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.65(s,1H),10.07(s,1H),7.76(s,1H),7.74(d,J＝1.6Hz,2H),7.68(t,J＝1.7Hz,1H),6.85–6.81(m,2H),6.51(dd,J＝2.0,0.8Hz,1H),5.45(s,1H),4.83(d,J＝13.0Hz,1H),4.42(d,J＝12.9Hz,1H),4.09(s,2H),2.82(d,J＝16.3Hz,1H),2.62(dd,J＝16.5,4.0Hz,1H),2.38(d,J＝14.7Hz,1H),2.22–2.14(m,1H),1.76(s,4H),1.23(d,J＝6.8Hz,5H),1.17(s,3H),1.03(s,3H),0.99(s,3H)。
Preparation of Compound II-5
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4694g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2046g of benzoyl hydrazine, and the mixture is stirred for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-5 as a white solid, 0.2760g, 47% yield. The molecular formula is: c33H36N2O8，HR-ESUMS m/z：589.2541[M+H]+，1H NMR(300MHz,DMSO-d6)δ10.92(s,1H),7.86(d,J＝7.4Hz,2H),7.74(s,1H),7.68(t,J＝1.7Hz,1H),7.61–7.55(m,1H),7.51(t,J＝7.3Hz,2H),6.52(s,1H),5.46(s,1H),4.85(d,J＝13.0Hz,1H),4.43(d,J＝13.0Hz,1H),4.10(d,J＝5.0Hz,2H),2.95(d,J＝14.2Hz,1H),2.82(d,J＝16.4Hz,1H),2.63(dd,J＝16.5,4.0Hz,1H),2.42(d,J＝14.8Hz,1H),2.19(d,J＝14.6Hz,1H),1.77(s,3H),1.25(d,J＝4.6Hz,3H),1.18(s,3H),1.04(s,3H),1.00(s,3H)。
Preparation of Compound II-6
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4707g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2258g of p-methyl benzoyl hydrazine, and the mixture is stirred for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-6 as a white solid, 0.3087g, 51% yield. The molecular formula is: c34H38N2O8，HR-ESUMS m/z：625.2526[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.77(s,1H),7.85(d,J＝8.4Hz,2H),7.74(d,J＝1.5Hz,1H),7.68(t,J＝1.8Hz,1H),7.04(d,J＝8.7Hz,2H),6.51(d,J＝1.6Hz,1H),5.45(s,1H),4.84(d,J＝13.1Hz,1H),4.43(d,J＝13.0Hz,1H),4.09(s,2H),3.83(s,3H),2.62(dd,J＝16.6,4.0Hz,1H),2.40(d,J＝14.9Hz,1H),2.18(d,J＝14.6Hz,1H),1.76(s,4H),1.22(s,3H),1.17(s,3H),1.03(s,3H),0.99(s,3H)。
Preparation of Compound II-7
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4704g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2268g of 4-carbamyl hydrazine, and stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-7 as a white solid, 0.3526g, 58% yield. The molecular formula is: c33H37N3O8，HR-ESUMS m/z：626.2480[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.41(s,1H),7.75–7.72(m,1H),7.68(t,J＝1.7Hz,1H),7.61(d,J＝8.6Hz,2H),6.57(d,J＝8.6Hz,2H),6.51(dd,J＝2.0,0.8Hz,1H),5.73(s,2H),5.45(s,1H),4.82(d,J＝13.0Hz,1H),4.42(d,J＝12.9Hz,1H),4.13(s,1H),3.17(d,J＝5.2Hz,0H),2.91(d,J＝14.4Hz,1H),2.81(d,J＝16.4Hz,1H),2.62(dd,J＝16.5,4.0Hz,1H),2.36(d,J＝14.7Hz,1H),2.17(d,J＝14.5Hz,1H),1.91(s,1H),1.75(d,J＝9.0Hz,3H),1.22(s,3H),1.17(s,3H),1.03(s,3H),0.98(s,3H)。
Preparation of Compound II-8
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4032g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2204g of 4-chlorobenzoyl hydrazine, and the mixture is stirred for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-8 as a white solid, 0.2721g, 51% yield. The molecular formula is: c33H35ClN2O8，HR-ESUMS m/z：645.1985[M+Na]+，1H NMR(300MHz,DMSO-d6)δ10.99(s,1H),7.89(d,J＝8.2Hz,2H),7.73(s,1H),7.67(t,J＝1.7Hz,1H),7.58(d,J＝8.1Hz,2H),6.51(s,1H),5.45(s,1H),4.85(d,J＝13.0Hz,1H),4.42(d,J＝13.0Hz,1H),4.09(s,2H),2.87(d,J＝15.3Hz,0H),2.76(d,J＝16.2Hz,1H),2.63(dd,J＝16.5,4.0Hz,1H),2.41(d,J＝14.9Hz,1H),2.18(d,J＝14.2Hz,1H),1.77(s,4H),1.24(d,J＝3.6Hz,4H),1.17(s,3H),1.04(s,3H),0.99(s,3H)。
Preparation of Compound II-9
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4011g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2147g of 3-chlorobenzoyl hydrazine, and stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-9 as a white solid, 0.2853g, 54% yield. The molecular formula is: c33H35ClN2O8，HR-ESUMS m/z：645.2021[M+Na]+，1H NMR (300MHz, DMSO-d6) δ 11.03(s,1H),7.90(s,1H),7.83(d, J ═ 7.4Hz,1H),7.74(s,1H),7.68(t, J ═ 1.7Hz,1H),7.64(s,1H),7.55(t, J ═ 7.8Hz,1H),6.51(s,1H),5.45(s,1H),4.85(d, J ═ 12.9Hz,1H),4.43(d, J ═ 12.9Hz,1H),4.09(s,2H),2.87(d, J ═ 15.0Hz,0H),2.76(d, J ═ 16.7Hz,1H),2.63(d, J ═ 16.5, 0, 3.42H), 3.3.3H, 3.7 (d, 1H), 3.3.7H, 1H), 3.3.3H, 3.3H, 3.5 (d, 1H), 3.7H, 1H, 3H. The molecular formula is:
preparation of Compound II-10
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4695g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.2079g of 2-pyrazine formylhydrazine, and the mixture is stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-3 as a white solid, 0.1375g, 23% yield. The molecular formula is: c31H34N4O8，HR-ESUMS m/z：613.2276[M+Na]+，1H NMR(300MHz,DMSO-d6)δ11.25(s,1H),9.20(d,J＝1.5Hz,1H),8.91(d,J＝2.5Hz,1H),8.79(dd,J＝2.5,1.5Hz,1H),7.75(s,1H),7.68(t,J＝1.8Hz,1H),6.52(d,J＝1.3Hz,1H),5.47(s,1H),4.84(d,J＝12.9Hz,1H),4.44(d,J＝13.4Hz,1H),4.11(d,J＝6.1Hz,2H),3.13(t,J＝15.1Hz,0H),2.91(d,J＝14.6Hz,1H),2.82(d,J＝16.3Hz,1H),2.65(d,J＝3.9Hz,1H),2.60(d,J＝3.1Hz,1H),2.45(s,0H),2.31(d,J＝3.1Hz,0H),2.26(d,J＝3.3Hz,0H),1.24(s,4H),1.18(d,J＝5.0Hz,4H),1.11(s,1H),1.05–0.99(m,7H)。
Preparation of Compound II-11
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4033g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.1712g of isoniazide, and the mixture is stirred for 48 hours at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-11 as a white solid, 0.1310g, 30% yield. The molecular formula is: c32H35N3O8，HR-ESUMS m/z：598.2541[M+H]+,1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),9.01(s,1H),8.21(d,J＝8.0Hz,1H),7.74(s,1H),7.68(t,J＝1.8Hz,1H),7.56(d,J＝7.1Hz,1H),6.52(s,1H),5.46(s,1H),4.85(d,J＝13.0Hz,1H),4.43(d,J＝13.0Hz,1H),4.13–4.07(m,2H),2.99(d,J＝14.6Hz,1H),2.82(d,J＝16.4Hz,1H),2.63(dd,J＝16.4,4.0Hz,1H),2.43(d,J＝14.9Hz,1H),2.18(d,J＝14.1Hz,1H),1.77(s,3H),1.21(d,J＝21.3Hz,9H),1.05(s,3H),1.00(s,4H)。
Preparation of Compound II-12
The compound limonin is taken as a starting material and is sequentially placed in a 50ml eggplant-shaped bottleAdding limonin 0.4061g, methanol 20ml, glacial acetic acid 0.200ml, 3-pyridine carbohydrazide 0.1702g, stirring at 70 deg.C for 48 h. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-12 as a white solid, 0.1128g, 30% yield. The molecular formula is: c32H35N3O8，HR-ESUMS m/z：598.2541[M+H]+，1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),9.01(s,1H),8.74(d,J＝4.8Hz,1H),8.20(d,J＝7.9Hz,1H),7.73(s,1H),7.67(t,J＝1.8Hz,1H),7.55(d,J＝7.0Hz,1H),6.51(s,1H),5.45(s,1H),4.84(d,J＝13.1Hz,1H),4.42(d,J＝13.0Hz,1H),4.17–4.00(m,2H),2.96(s,0H),2.81(d,J＝16.4Hz,1H),2.62(dd,J＝16.4,4.0Hz,1H),2.50(p,J＝1.9Hz,9H),2.19(s,1H),1.76(s,3H),1.38–1.29(m,2H),1.24(d,J＝6.2Hz,3H),1.18(s,5H),1.04(s,3H),0.99(s,4H)。
Preparation of Compound II-13
The compound limonin is used as a starting material, 50ml of eggplant-shaped bottles are sequentially added with the limonin, 0.4717g, 20ml of methanol, 0.200ml of glacial acetic acid and 0.3062g of 4-trifluoromethyl benzoyl hydrazine, and stirred for 48h at 70 ℃. TLC detection till the reaction is complete, cooling and decompressing to evaporate the solvent. The crude product is extracted by dichloromethane: column chromatography with methanol (100:1) gave II-13 as a white solid, 0.1908g, 30% yield. The molecular formula is: c34H35F3N2O8，HR-ESUMS m/z：657.1222[M+H]+，1H NMR(300MHz,DMSO-d6)δ11.14(s,1H),8.06(d,J＝8.0Hz,2H),7.90(d,J＝8.1Hz,2H),7.74(s,1H),7.67(t,J＝1.7Hz,1H),6.52(s,1H),5.46(s,1H),4.86(d,J＝13.0Hz,1H),4.43(d,J＝13.0Hz,1H),4.10(s,2H),2.94(d,J＝14.4Hz,1H),2.82(d,J＝16.4Hz,1H),2.66(d,J＝4.0Hz,1H),2.60(d,J＝3.9Hz,0H),2.19(d,J＝14.6Hz,1H),1.78(s,4H),1.24(s,4H),1.18(s,3H),1.04(s,4H),1.02–0.99(m,3H)。
Example 2
Taking 0.1g of the compound prepared in the example 1, 0.5g of starch, 0.05g of talcum powder and a proper amount of CMC-Na as a wetting agent, granulating and tabletting.
Example 3
The application of the limonin compound as a medicine for treating echinococcosis:
the experimental contents are as follows:
1.1 pharmaceutical intervention for in vitro culture of Echinococcus granulosus Procoenuruses
Separating echinococcus granulosus prototheca in vitro, performing aseptic culture in a 24-well plate, adding limonin and limonin derivatives with different concentrations, intervening by positive drug albendazole, and dividing into a solvent group (0.1% DMSO control group), a limonin group (80 mu M), an obacunone group (80 mu M), a derivative group (80 mu M) and an albendazole group (40 mu M) for intervening for 0 day, 3 days and 5 days respectively, observing the drug action change of the larvae, and observing the activity of the prototheca by using eosin. The results are shown in FIG. 1 and FIG. 2, which are eosin staining patterns of limonoids and albendazole, a positive drug, for 3 days and 5 days of in vitro incubation of echinococcus granulosus metacercaria, respectively, wherein a: blank group, b: albendazole, c: limonin, d: compound II-5, e: compound II-10, f: the compound II-13 can see that the metacercaria treated by the limonin and the derivatives thereof has expanded volume, eversion of head nodes and abnormal overall shape, the in vitro inhibition rate after treatment is shown in figure 3 after eosin dyeing, the in vitro anti-echinococcosis activity of the limonin is superior to that of the albendazole treatment group, the synthesized part of the limonin derivatives presents a metering dependence relation on the anti-echinococcosis activity, and the experimental result shows that the limonin and the derivatives thereof have treatment effect on echinococcosis in vitro.
1.2 pharmacological intervention in vivo on Echinococcus granulosus infected mice
1.2.1 model inoculation
(1) Carefully treating hydatid cyst of a patient infected with echinococcosis with 75% ethanol, transferring into a biological safety cabinet, carefully absorbing cyst fluid into a sterile beaker by using a 50ml sterile injector, standing until protocephalic segments naturally settle, discarding supernatant, washing three times by using sterile PBS (phosphate buffer solution), adjusting the density of protocephalic segments to 10000 protocephalic segments/ml, performing intraperitoneal injection on 0.2ml protocephalic segment suspension in each KM mouse, infecting for 150 days, randomly taking the mouse, dissecting, and judging whether molding is successful or not.
(2) Pharmacodynamic tests are carried out on mice successfully molded, and the experiments are divided into a blank group, a positive drug group (albendazole group) and a treatment group (limonin group), and the three groups are total, and the administration dosage is 50 mg/kg/day and 15 days.
The specific implementation is as follows: the albendazole group and the limonin group were administered orally in a volume of 0.2ml, with the drug being 50 mg/kg/day per mouse daily intake, after grinding in a mortar with 0.4% CMC-Na. After continuously administering for 15 days, the mice are euthanized, cysts are collected from the abdominal cavity, the damp weight of the cysts is measured, the inhibition rate of the drug to the cysts is calculated, the inhibition rate is (blank group cyst damp weight-treatment group cyst damp weight)/blank group cyst damp weight is 100%, the weight and the damp weight of the mice are recorded, the liver, the kidney and the cysts are taken to be pathological sections, and the conditions of the change of the kidney and the liver, the thickness of the cyst wall, the thickness of a germinal layer and the like are observed. Finally, statistical analysis is carried out by using a sps 20.0, data are expressed by mean values +/-standard deviation, One-way ANOVA analysis is adopted among groups, and the p value is smaller than 0.05 and has statistical difference.
(3) The liver, kidney and cyst of each group of mice were fixed with 4% paraformaldehyde, embedded into sections, and after H & E staining, the liver, kidney and cyst of each group of experimental groups were observed.
As shown in fig. 4, wherein a: a normal group; b: a model group; c: albendazole treatment group; d: limonin treatment group; the limonin group has normal liver group morphology, liver tissue cell parts of albendazole lose characteristic cell structures, and the limonin has certain liver protection effect and can repair damaged liver cells, and the albendazole has liver damage. As shown in fig. 5, wherein a: a normal group; b: a model group; c: albendazole treatment group; d: in the limonin treatment group, the kidney tissue cell parts of the limonin group and the albendazole lose characteristic cell structures and have certain damage.
As shown in fig. 6, the results of h.e staining of mouse cyst tissue after intervention of limonin and albendazole with echinococcus granulosus metacercaria, where a: model group, b: albendazole treatment group, c: in the limonin treatment group, the echinococcus capsularis tissues treated by the limonin are obviously changed, the horny layer is thinned or disappears, a large amount of large-area vacuoles appear in the whole outer membrane, the structure is seriously changed, and germinal layer cells are thinned or disappear; therefore, the limonin compound can effectively treat echinococcosis and has excellent insecticidal effect
TABLE 1 cyst Wet weight and inhibition Rate in groups of mice after 3 weeks of treatment
Figure BDA0003056152600000111
As shown in fig. 7, fig. 8 and table 1, the wet weight of the cysts of the mice in each group after 3 weeks of treatment was significantly reduced compared to the model group (4.1955 ± 2.7995g) in the albendazole group (0.9480 ± 0.5558g) and the limonin group (0.2037 ± 0.2259g), and the cysts of the mice in vivo almost disappeared (p ═ 0.017 and p ═ 0.03), and compared to the limonin group and the positive albendazole group, the therapeutic effect of the limonin was significantly higher than that of the albendazole group (p ═ 0.021), and the inhibition rate reached 95%.
The limonin and the derivatives thereof are novel natural compounds which can be used for developing anti-echinococcosis drugs, in-vivo pharmacodynamic data suggest that the limonin is a relatively efficient anti-echinococcosis drug molecule, and key members of the signal path can also be used as targets for screening novel echinococcosis drugs. The invention can provide a new way for treating echinococcosis.

Claims (7)

1. A limonin compound is characterized by having a general formula shown as I, II or III:
Figure FDA0003056152590000011
wherein X is-O-, -CH2-, Bond;
R1is C1-C6 alkyl,
Figure FDA0003056152590000012
Figure FDA0003056152590000013
Figure FDA0003056152590000014
One of (1);
R2all C1-C4 alkyl, -X, -CX substituted by ortho-para3、-OCH3、-NH2、-NO2、
Figure FDA0003056152590000015
-one of OH;
R3all alkyl groups of C1-C2, ═ CH2One kind of (1).
2. A process for the preparation of limonoids of formula II as claimed in claim 1, comprising the steps of:
Figure FDA0003056152590000016
wherein the reaction reagent is alkyl hydrazine or acyl hydrazine, and the reaction solvent is methanol or ethanol solution.
3. A process for the preparation of limonoids of formula III as claimed in claim 1, including the steps of:
Figure FDA0003056152590000017
wherein the reaction reagent is iodoalkane or pyrrolidine, and the solvent is ethanol or benzene solution.
4. The method of claim 2, wherein the limonoid of formula II is prepared from the limonoid of formula I using acetic acid as the catalyst.
5. The use of a limonoid according to claim 1 as a medicament for the treatment of echinococcosis.
6. The use of claim 5, further comprising a pharmaceutically acceptable salt of a limonoid with an acid and a pharmaceutically acceptable carrier.
7. The use according to claim 6, wherein the acid is one or more of hydrogen chloride, hydrogen bromide, sulphuric acid, carbonic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulphonic acid, benzenesulphonic acid, p-toluenesulphonic acid or ferulic acid.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022237913A1 (en) * 2021-05-08 2022-11-17 南京医科大学 Limonin compound, preparation method therefor, and application of limonin compound as drug for treating hydatidosis/echinococcosis

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117327028B (en) * 2023-09-25 2024-07-23 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心) Phenothiazine derivative, pharmaceutical composition and application thereof in treating echinococcosis

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101367823A (en) * 2008-09-25 2009-02-18 成都普思生物科技有限公司 Method for separating citrate, evodiamine and rutaecarpine from evodia rutaecarpa
CN103588854A (en) * 2013-11-14 2014-02-19 中国药科大学 Limonin oxime ether derivatives as well as preparation methods and medical applications thereof
CN105481878A (en) * 2015-12-31 2016-04-13 胡文杰 Novel limonin compound and preparation method and medical application thereof
CN108727461A (en) * 2018-06-25 2018-11-02 中国药科大学 A method of detaching limonoid from Dictamnus angustifolius skin
CN111574533A (en) * 2020-05-26 2020-08-25 中国药科大学 Limonin A ring-opening aminated derivative or pharmaceutically acceptable salt thereof, preparation method and application
WO2020181173A1 (en) * 2019-03-07 2020-09-10 Kelley Darshan Singh Limonoid compounds for treatment of cancer

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113234089B (en) * 2021-05-08 2022-07-15 南京医科大学 Limonin compound, preparation method and application thereof as medicine for treating echinococcosis

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101367823A (en) * 2008-09-25 2009-02-18 成都普思生物科技有限公司 Method for separating citrate, evodiamine and rutaecarpine from evodia rutaecarpa
CN103588854A (en) * 2013-11-14 2014-02-19 中国药科大学 Limonin oxime ether derivatives as well as preparation methods and medical applications thereof
CN105481878A (en) * 2015-12-31 2016-04-13 胡文杰 Novel limonin compound and preparation method and medical application thereof
CN108727461A (en) * 2018-06-25 2018-11-02 中国药科大学 A method of detaching limonoid from Dictamnus angustifolius skin
WO2020181173A1 (en) * 2019-03-07 2020-09-10 Kelley Darshan Singh Limonoid compounds for treatment of cancer
CN111574533A (en) * 2020-05-26 2020-08-25 中国药科大学 Limonin A ring-opening aminated derivative or pharmaceutically acceptable salt thereof, preparation method and application

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AMMAR SELLES SIDI MOHAMMEDL ET AL.,: "IN-VITRO SCOLICIDAL ACTIVITY OF CITRUSLIMON EUREKA JUICE", 《AGRICULTURA》 *
COLUMBUS, OHIO, US，CHEMICAL ABSTRACT SERVICE: "STN Registry", 《STN检索报告》 *
RUNYU YANG ET AL.,: "Limonin Attenuates LPS-Induced Hepatotoxicity by Inhibiting Pyroptosis via NLRP3/Gasdermin D Signaling Pathway", 《J. AGRIC. FOOD CHEM.》 *
姜文霞 等编: "《病理学》", 31 July 2020, 华中科技大学出版社 *
张 贝 等: "柠檬苦素的性质及其生物活性研究进展", 《农产品加工》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022237913A1 (en) * 2021-05-08 2022-11-17 南京医科大学 Limonin compound, preparation method therefor, and application of limonin compound as drug for treating hydatidosis/echinococcosis

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