CN111233629A

CN111233629A - Method for preparing compound BZ-92 from traditional Chinese medicine bighead atractylodes rhizome

Info

Publication number: CN111233629A
Application number: CN202010168202.2A
Authority: CN
Inventors: 谢治深; 高改; 吴丽敏; 李二稳; 王梦瑶; 赵建平
Original assignee: Henan University of Traditional Chinese Medicine HUTCM
Current assignee: Henan University of Traditional Chinese Medicine HUTCM
Priority date: 2020-03-11
Filing date: 2020-03-11
Publication date: 2020-06-05
Anticipated expiration: 2040-03-11
Also published as: CN111233629B

Abstract

The invention discloses a method for preparing a compound BZ-92 from a traditional Chinese medicine bighead atractylodes rhizome. The compound BZ-92 is prepared by separating the bighead atractylodes rhizome serving as the raw material for the first time, and the advantage of abundant resources of the bighead atractylodes rhizome is fully exerted. The prior art does not report that the compound BZ-92 is contained in the largehead atractylodes rhizome, and does not report that other plants in the compositae of the largehead atractylodes rhizome contain the compound BZ-92. The invention achieves breakthrough achievement in the preparation of the compound BZ-92.

Description

Method for preparing compound BZ-92 from traditional Chinese medicine bighead atractylodes rhizome

Technical Field

The invention belongs to the field of natural medicinal chemistry, relates to a separation preparation method of a known compound, and particularly relates to a method for preparing a compound BZ-92 from a traditional Chinese medicinal material, namely bighead atractylodes rhizome.

Background

Compound BZ-92 is a natural product having the chemical structure:

the compound BZ-92 is found to have various pharmacological activities.

However, the prior art has been less studied on the isolation and preparation of the compound BZ-92. The invention is especially provided for expanding the separation raw material selection and separation and purification process of the compound BZ-92.

The atractylodes macrocephala koidz is a perennial herb of the Compositae, is also called Zhejiang kou, Dongzhu, Yushu and the like, is used as a medicine by using roots and stems, has the effects of stopping sweating, preventing miscarriage, eliminating dampness and promoting diuresis, strengthening spleen and replenishing qi, is the most commonly used medicine in a traditional Chinese medicine prescription, and has the theory of 'ten medicines and nine technologies'. At present, China has a relatively large-scale rhizoma atractylodis macrocephalae planting industry, and the rhizoma atractylodis macrocephalae resources are extremely rich.

At present, no method for preparing the compound BZ-92 by taking the largehead atractylodes rhizome as a raw material exists.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a method for preparing a compound BZ-92 from a traditional Chinese medicine bighead atractylodes rhizome.

The above purpose of the invention is realized by the following technical scheme:

a method for separating and preparing a compound BZ-92 has the following chemical structure, and uses a Chinese medicinal material of largehead atractylodes rhizome as a separation raw material;

further, the separation preparation step comprises:

step S1, crushing the traditional Chinese medicine rhizoma atractylodis macrocephalae, extracting with an alcohol-water solution, and concentrating an extracting solution to obtain a crude extract;

step S2, suspending the crude extract obtained in the step S1 with a proper amount of water, extracting with ethyl acetate, collecting ethyl acetate extract, and concentrating to obtain an ethyl acetate extract;

step S3, separating the ethyl acetate extract obtained in the step S2 by normal phase silica gel column chromatography, monitoring a compound BZ-92 in eluent, and combining the eluent containing the compound BZ-92 to obtain a compound BZ-92 enrichment substance; separating the enriched BZ-92 with reverse phase silica gel column chromatography, monitoring the compound BZ-92 in the eluent, combining the eluent containing the compound BZ-92, concentrating and drying to obtain the compound BZ-92.

Further, the alcohol aqueous solution in step S1 is 95% ethanol.

Furthermore, the elution solvent of the normal phase silica gel column chromatography in step S3 is a dichloromethane-methanol mixed solvent, and the volume ratio and the elution amount of the two mixed solvents are sequentially: 500:1, 6 column volumes; 100:1, 4 column volumes; 50:1, 6 column volumes.

Furthermore, the monitoring method of normal phase silica gel column chromatography is thin layer chromatography.

Further, the elution solvent of the reverse phase silica gel column chromatography in step S3 was 85% methanol, and the elution was performed isocratically.

Further, the monitoring method of the reversed phase silica gel column chromatography is HPLC method.

Has the advantages that:

the compound BZ-92 is prepared by separating the bighead atractylodes rhizome serving as the raw material for the first time, and the advantage of abundant resources of the bighead atractylodes rhizome is fully exerted. The prior art does not report that the compound BZ-92 is contained in the largehead atractylodes rhizome, and does not report that other plants in the compositae of the largehead atractylodes rhizome contain the compound BZ-92. The invention achieves breakthrough achievement in the preparation of the compound BZ-92.

Drawings

FIG. 1 shows the compound BZ-92¹HNMR spectra (400MHz, deuterated chloroform);

FIG. 2 shows the compound BZ-92¹³CNMR spectra and DTPT135 spectra (100MHz, deuterated chloroform).

Detailed Description

The following detailed description of the present invention is provided in connection with the accompanying drawings and examples, but not intended to limit the scope of the invention.

Example 1: preparation and structure confirmation of compound BZ-92

The preparation method comprises the following steps:

step S1, crushing 2kg of Chinese herbal medicine rhizoma atractylodis macrocephalae, soaking and extracting with 95% ethanol (5LX 3 times), combining extracting solutions, and concentrating under pressure to obtain 200g of crude extract;

step S2, suspending the crude extract obtained in step S1 with 2L of water, extracting with ethyl acetate (2LX 3 times), collecting ethyl acetate extract, and concentrating to obtain ethyl acetate extract;

step S3, eluting the ethyl acetate extract (100g) obtained in step S2 with dichloromethane (500: 1(6 column volumes), 100:1(4 column volumes) and 50:1(6 column volumes) in sequence by normal phase silica gel column chromatography, combining eluates containing the compound BZ-92 according to TLC analysis, and concentrating to obtain an enriched compound BZ-92 Fr 3; then, the enriched product Fr3(5.0g) of the compound BZ-92 was eluted with 85% methanol water isocratic by reverse phase silica gel column chromatography, the compound BZ-92 in the eluate was monitored by HPLC, the eluates containing the compound BZ-92 were combined, concentrated and dried to obtain BZ-92 (about 200mg, purity 98%).

And (3) structure confirmation: HR-ESI-MS shows [ M + H [ ]]+ is m/z 221.1563, the molecular formula C is obtained by combining the nuclear magnetic characteristics₁₅H₂₄O, unsaturation degree is 4. Hydrogen nuclear magnetic resonance spectroscopy data δ h (ppm, CDCl)₃H-1 α (1.32, m), H-1 β (1.58, m), H-2 β (1.23, m), H-2 β (1.36, m), H-3 α (2.38, br d, J ═ 13.0Hz), H-3 β (2.1l, m), H-5(2.11, br t, J ═ 11.0Hz), H-6 α (1.90, m), H-6 β (2.01, m), H-8(5.67, dd, J ═ 7.0,3.0Hz), H-9 α (1.60, m), H-9 β (1.65, m), H-12(1.36, s), H-13(1.37, s), H-14(0.69, s), H- α (15, 6, 7.61, 15, 7.82, 25, g), C NMR (82 ppm), H-12 (CDS), H-13 (2, M), H-3.69, M), H-14 (2, M), H-3.6, H-3₃,100MHz):41.8(CH₂，1-C)，23.6(CH₂,2-C)，37.2(CH₂,3-C)，150.3(C,4-C)，44.9(CH,5-C)，24.4(CH₂,6-C)，142.4(C,7-C)，116.9(CH，8-C)，41.6(CH₂,9-C)，34.1(C,10-C)，72.9(C,11-C)，29.0(CH₃，12-C)，29.0(CH₃,13-C)，17.2(CH₃,14-C)，106.3(CH₂15-C). FIG. 1 shows the compound BZ-92¹HNMR spectra (400MHz, deuterated chloroform); FIG. 2 shows the compound BZ-92¹³CNMR and DTPT135 spectra(100MHz, deuterated chloroform). The chemical structure is confirmed as follows:

The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.

Claims

1. A method for separating and preparing a compound BZ-92, wherein the chemical structure of the compound BZ-92 is as follows: taking a Chinese medicinal material of largehead atractylodes rhizome as a separation raw material;

2. the method of claim 1, wherein the step of separately preparing comprises:

3. The method of claim 2, wherein: the alcohol aqueous solution in step S1 is 95% ethanol.

4. The method according to claim 2, wherein the elution solvent of the normal phase silica gel column chromatography in step S3 is a dichloromethane-methanol mixed solvent, and the volume ratio and the elution amount of the two mixed solvents are, in order: 500:1, 6 column volumes; 100:1, 4 column volumes; 50:1, 6 column volumes.

5. The method of claim 4, wherein: the monitoring method of normal phase silica gel column chromatography is thin layer chromatography.

6. The method of claim 2, wherein: the elution solvent of the reversed phase silica gel column chromatography in the step S3 is 85% methanol, and isocratic elution is carried out.

7. The method of claim 6, wherein: the monitoring method of the reversed phase silica gel column chromatography is HPLC method.