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Ethanol extract of Dulong Chonglou and its application in pharmaceuticals and its chemical constituents separation and identification method

A technology of ethanol extraction and heavy building, applied in chemical instruments and methods, plant/algae/fungus/moss components, organic chemistry, etc.

Active Publication Date: 2019-08-02
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the medicinal aspect of Dulong Chonglou. The present invention finds that the ethanol extract of Dulong Chonglou (PDE4) and its total saponins (PDE5) have significant cytotoxic activity on the basis of previous activity screening. Therefore, Clarifying the chemical substance basis of PDE4 and PDE5 is of great significance for the establishment of quality control standards for Dulongzhonglou, PDE4 or PDE5, and the development of new drugs based on Dulongzhonglou, PDE4 or PDE5

Method used

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  • Ethanol extract of Dulong Chonglou and its application in pharmaceuticals and its chemical constituents separation and identification method
  • Ethanol extract of Dulong Chonglou and its application in pharmaceuticals and its chemical constituents separation and identification method

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Experimental program
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Effect test

Embodiment 1

[0026] The preparation of PDE4 of the present invention and PDE5 and the separation of contained chemical composition thereof:

[0027] 1 kg of dried rhizome of Dulong Zhonglou, crushed, extracted with 3-4 liters of 90%-95% ethanol at 60-65°C for 3-4 hours, filtered to obtain filtrate and filter residue, and the filter residue was extracted twice according to the above conditions. Combine the filtrates obtained from the three extractions, and concentrate to obtain 133 g of the ethanol extract of Dulong Chonglou (referred to as PDE4).

[0028] Add 1 liter of water to PDE4 to make a suspension, and extract 3 times with 1 liter of petroleum ether, chloroform, and n-butanol successively. After the extract was concentrated, petroleum ether extract (2 g), chloroform extract (30 g) and n-butanol extract (30 g) were obtained respectively. The n-butanol extraction part is the total saponins of Dulong Chonglou, which is recorded as PDE5.

[0029] Segment PDE5 (30g) with 80-100 mesh si...

Embodiment 2

[0034] Structural identification of the chemical components isolated from the total saponins (PDE5) of the present invention:

[0035] Pachyphyll saponin V (PDE10), white solid, molecular formula C 39 h 62 o 12 , CAS registry number 19057-67-1. 1 H NMR (C 5 D. 5 N, 500MHz) δ: 6.40 (1H, br s), 5.29 (1H, br d, J = 5.1Hz), 1.78 (3H, d, J = 6.3Hz), 1.12 (3H, d, J = 7.0Hz) , 1.04 (3H, s), 0.81 (3H, s), 0.68 (3H, d, J=5.5Hz). 13 C NMR (C 5 D. 5 N, 126MHz) δ: 140.9, 121.8, 109.3, 102.2, 100.4, 81.1, 79.7, 78.4, 77.9, 77.8, 74.2, 72.9, 72.7, 71.8, 69.6, 66.9, 62.9, 62.7, 56.6, 50.3, 42.0, 40.5, 39.9, 39.0, 37.5, 37.2, 32.3, 32.3, 31.8, 31.7, 30.6, 30.2, 29.3, 21.1, 19.5, 18.8, 17.4, 16.4, 15.1. ESIMS m / z 721[M-H] - . Its NMR data are basically consistent with the data in the literature [Li Zhongqiong, Lin Ruichao, Fu Wen, Wang Gangli, Wei Feng, Hang Taijun. Research on the Chemical Components of Lettuce Flowers. Chinese Herbal Medicine, 2001,32(2):101-104].

[0036] Papaya...

Embodiment 3

[0042] The inhibitory effect of ethanol extract (PDE4), n-butanol extraction fraction (PDE5) and compounds isolated from PDE5 on the growth of cancer cells:

[0043] PDE4, PDE5 and the compounds PDE10, PDE11, PDE12, PDE14, PDE15 and PDE22 isolated from PDE5 were tested for their cytotoxic activity, and the results are shown in Table 1. The results showed that PDE4, PDE5, PDE10, PDE11, PDE14 and PDE15 had significant inhibitory activity on leukemia HL-60 cells, liver cancer SMMC-7721 cells, lung cancer A-549 cells, breast cancer MCF-7 cells and colon cancer SW480 cells, Its activity is comparable to the positive control drug cisplatin.

[0044] Table 1 Inhibitory effect of Dulong Chonglou extract and compounds on cancer cell growth

[0045]

[0046]

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Abstract

The invention discloses an ethanol extract (PDE4) and total saponin (PDE5), which have anticancer activity, from paris dulongensis H. Li&Kurita, preparation methods of the ethanol extract (PDE4) and the total saponin (PDE5) and application of the ethanol extract (PDE4) and the total saponin (PDE5) to the preparation of a medicine for treating or preventing leukemia, a liver cancer, a lung cancer, a mammary cancer and a colon cancer, and provides a separation and identification method of chemical components in the PDE 5. Polyphyllin V, polyphyllin I, beta-ecdysone, protodioscin, parisaponin I and ginsenoside Rg1 are separated from the PDE5 and identified by adopting a separation method of chromatographic column chromatography and semi-preparative high performance liquid chromatography and an identification method of a mass spectrum and a nuclear magnetic resonance wave spectrum. The separation and identification method is used for clarifying main chemical components of the PDE5 for the first time, can be used for simply ad quickly obtaining the chemical components from the PDE5 through a provided method, and is used for reconstructing a quality control standard for the paris dulongensis H. Li&Kurita, the extract PDE4 of the paris dulongensis H. Li&Kurita or the total saponin PDE5 of the paris dulongensis H. Li&Kurita.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of medicine and the field of natural product extraction and separation technology, and in particular relates to the ethanol extract PDE4 of Dulongzhonglou, the total saponins PDE5 of Dulongzhonglou and their preparation methods, as well as the medicines for treating cancer, and the anti-cancer of Dulongzhonglou A method for the isolation and identification of the chemical constituents of the active site PDE5. [0003] Background technique: [0004] Plants of the genus Paris in the Melanthiaceae family have high medicinal value, among which Paris polyphylla var.yunnanensis (Franch.) Hand.-Mazz and Paris polyphylla var. chinensis (Franch.) Hara] was included in the 2015 edition of "Pharmacopoeia of the People's Republic of China". Great progress has been made in the artificial planting of Chonglou plants. For example, in the Nujiang Lisu Autonomous Prefecture of Yunnan Province, the planting area of ​​...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J71/00C07J9/00C07J17/00A61K31/7048A61K31/575A61K36/896G01N24/08G01N30/02G01N30/90A61P35/00A61P35/02
CPCA61K31/575A61K31/7048A61K36/896C07J9/00C07J17/005C07J71/0005G01N24/087G01N30/02G01N30/90
Inventor 王跃虎汉秋燕李晓艳左何英杨珺夏梦媛罗吉凤李恒杨永平
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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