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The method and application of extracting n6-(2-hydroxyethyl)adenosine from Cordyceps militaris

A technology of hydroxyethyl and Cordyceps militaris is applied in chemical instruments and methods, medical preparations containing active ingredients, pharmaceutical formulations, etc., and can solve the problem of large dosage, no development of anti-insomnia active monomers, long taking time, etc. problem, to achieve the effect of improving extraction rate, reducing autonomous activity, and high consistency

Active Publication Date: 2015-12-30
贵州贵安精准医学股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these natural medicines have not developed anti-insomnia active monomers, so they have disadvantages such as large dosage and long taking time.

Method used

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  • The method and application of extracting n6-(2-hydroxyethyl)adenosine from Cordyceps militaris
  • The method and application of extracting n6-(2-hydroxyethyl)adenosine from Cordyceps militaris
  • The method and application of extracting n6-(2-hydroxyethyl)adenosine from Cordyceps militaris

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Extraction of N6-(2-hydroxyethyl)adenosine in Cordyceps militaris

[0041] 1. Raw materials and materials:

[0042] The fruiting bodies of Cordyceps militaris were produced in this laboratory.

[0043] 2. Reagents:

[0044]Ultrapure water was produced by Millipore Water Purifier; 95% ethanol was purchased from Tianjin Beifang Tianyi Chemical Reagent Factory; chromatography-grade methanol was purchased from Honeywell Company.

[0045] 3. Instruments and equipment:

[0046] The ultrafine pulverizer was purchased from Shandong Sanqing Stainless Steel Equipment Co., Ltd.; the desktop refrigerated centrifuge was purchased from Thermo Company; the rotary evaporator was purchased from Shanghai Yarong Biochemical Instrument Factory; C18 chromatographic column (150×250mm) Jiangsu Hanbang Technology Co., Ltd.; Multifunctional Membrane Separation System Hangzhou Kaijie Membrane Separation Technology Co., Ltd.

[0047] The extraction steps of N6-(2-hydroxyethyl)adeno...

Embodiment 2

[0056] Example 2: Confirmation of N6-(2-hydroxyethyl)adenosine

[0057] 1. Purity analysis by HPLC:

[0058] The target component N6-(2-hydroxyethyl)adenosine obtained in Example 1 was analyzed for purity by HPLC. The purity reached 98% through high performance liquid phase detection, and the HPLC spectrogram is shown in image 3 .

[0059] 2. Mass spectrometry analysis:

[0060] Positive ion mode electrospray mass spectrometry (ESI-MS) analysis of the target component N6-(2-hydroxyethyl)adenosine: Varian450GC-320TQ-M, Hanmeng Biotechnology (Tianjin) Co., Ltd., see the mass spectrum Figure 4 . Electrospray mass spectrometry [(+)-ESI-MS] gives a quasi-molecular ion peak m / z of 311.97 [M+H] + ; Prompt molecular composition is C 12 h 17 N 5 o 5 .

[0061] 3. NMR analysis:

[0062] NMR analysis of the target component N6-(2-hydroxyethyl)adenosine:

[0063] 1D 1 The data results in the HNMR spectrum (DMSO-d6, 400MHz) are as follows: δ: 8.358 (1H, s, H-8), 8.216 (1H, s...

Embodiment 3

[0067] Embodiment 3: N6-(2-hydroxyethyl) adenosine in anti-insomnia effect test

[0068] 1. Drugs and reagents:

[0069] Zaoren Anshen Capsules were purchased from Guizhou Tongjitang Pharmaceutical Co., Ltd.; Estazolam Tablets were purchased from Tianjin Pacific Pharmaceutical Co., Ltd.; pentobarbital sodium was packaged by Sigma.

[0070] 2. Test animals:

[0071] KM mice (18-22 g) (animal license number: SCXK (Beijing) 2012-0001, certificate number: 11400700020779) were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

[0072] 3. Equipment and instruments

[0073] The syringe was purchased from Shanghai Zhiyu Medical Instrument Co., Ltd.; the stopwatch and EL electronic balance were purchased from Changzhou Tianzhiping Instrument Equipment Co., Ltd.

[0074] Get the N6-(2-hydroxyethyl)adenosine that embodiment 1 extracts, carry out anti-insomnia effect test, method step is as follows:

[0075] 1. Drug preparation:

[0076] N6-(2-hydroxyethyl...

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Abstract

The invention provides a method for extracting N6-(2-ethoxyl) adenosine from cordyceps militaris. The method comprises the following steps: carrying out superfine comminution on cordyceps militaris fruiting body to obtain superfine powder; adding ultrapure water into the superfine powder, heating and extracting at the temperature of 100 DEG C, centrifuging, taking and drying the supernatant; adding a little ultrapure water to dissolve, adding 95% ethyl alcohol till the final concentration is 80%, carrying out ethanol precipitation, collecting and drying the supernatant overnight; dissolving by using a 15% methanol-water liquid till the concentration is 200mg / ml, separating through a preparative liquid chromatography, carrying out isocratic elution for 30-80 minutes by using the 15% methanol-water liquid, collecting the target chromatographic peak, concentrating and drying to obtain the N6-(2-ethoxyl) adenosine. The method is simple, controllable and good in repeatability; the purity of the extracted N6-(2-ethoxyl) adenosine can be up to 98%; and the N6-(2-ethoxyl) adenosine is proved to have the effect of resisting insomnia and can be used for preparing insomnia-resisting drugs.

Description

technical field [0001] The invention relates to the technical field of biological product processing, in particular to a method for extracting N6-(2-hydroxyethyl)adenosine from Cordyceps militaris, and the extracted N6-(2-hydroxyethyl)adenosine is used in the preparation Application of anti-insomnia drugs. Background technique [0002] Insomnia is a common disease, frequently-occurring disease, and has had a strong impact on people's life, work and physical and mental health. The incidence of serious accidents and traumas in long-term insomniacs is significantly higher than that in normal sleepers. Therefore, many medicines have been invented to treat insomnia. The barbiturates used in the early days had large side effects and strong dependence, and are now rarely used. Benzodiazepines have become the most commonly used sleeping pills in China because of their better curative effect, relatively safe, less respiratory depression, less drug interactions, and relatively diff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H19/167C07H1/08A61K31/7076A61P25/20
Inventor 张耀洲陈玉皎崔今松舒特俊陈剑清
Owner 贵州贵安精准医学股份有限公司
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