CN110893203A

CN110893203A - Double-order nux vomica processed product extract and preparation method thereof

Info

Publication number: CN110893203A
Application number: CN201911021646.7A
Authority: CN
Inventors: 秦伟瀚; 阳勇; 李卿; 王云红; 郭延垒
Original assignee: Chongqing Academy of Chinese Materia Medica
Current assignee: Chongqing Academy of Chinese Materia Medica
Priority date: 2019-10-25
Filing date: 2019-10-25
Publication date: 2020-03-20

Abstract

The invention provides a double-order nux vomica processed product extract, wherein the active ingredients of the double-order nux vomica processed product extract are strychnine and alkaloid generated by the reaction of strychnine and phenolic or weakly acidic ingredients in ginger. The double-stage processed nux vomica product extract provided by the invention overcomes the defect that a small amount of strychnine and strychnine is remained in the nux vomica extract; the volume is reduced, and conditions are created for the formulation development of the processed products of nux vomica.

Description

Double-order nux vomica processed product extract and preparation method thereof

Technical Field

The invention relates to a traditional Chinese medicine extract, in particular to a nux vomica concocted product extract.

Technical Field

Semen Strychni is dried mature seed of Strychni Pulveratum (S t r y h o n S nux-vomica L.) and semen Strychni (S.Wallichiana Steudex D.C.: S.p ieriana.). Contains alkaloid 2-5%, mainly strychnine and strychnine. Strychnos nux-vomica is bitter in taste, cold in nature and toxic. It enters liver and spleen meridians. Has effects in dispersing pathogen accumulation, relieving swelling, dredging meridians, and relieving pain; is used for treating rheumatism, numbness, paralysis, carbuncle, swelling and pain, and traumatic injury. In the records of Yi Xue Zhong Can xi Lu, nux vomica is superior to other herbs in that it can open meridians and collaterals and reach joints thoroughly. The clinical application of nux vomica is not declined for nearly a thousand years, which shows that the nux vomica has obvious curative effect and can be called 'Duyaqiang agent good for Jiechu'. But at the same time, because it is a toxic drug, the clinical application of the medicine is greatly limited.

The therapeutic dose of raw nux vomica is very close to the toxic dose, so the nux vomica needs to be processed firstly. The processing of Chinese herbs has been done by sand-scalding, frying, processing with licorice, soaking in vinegar, and burst-pressing, and the common purpose is to reduce toxicity. Modern multiple toxicological experimental studies prove that the various processing methods have exact attenuation function. Studies by chenchenchenille et al show that: the processing has certain influence on strychnine and strychnine content. The toxicity can be reduced by processing with vinegar, frying and processing with liquorice; the content of strychnine and strychnine in vinegar-soaked products is greatly reduced, the toxicity is reduced, but the titer can also be reduced. The sand scald product contains higher amount of strychnine and strychnine than the raw product. Ding's and the like process the raw nux vomica by using a popcorn machine, which proves that the content of strychnine is lower by a bursting method compared with a sand ironing method, thereby improving the clinical medication safety and curative effect of the processed nux vomica product. Chen Shi et al used 4 different herba Ephedrae preparata methods to process semen Strychni, determined the contents of strychnine and strychnine in different processed semen Strychni products, and conducted acute toxicity, analgesic effect and antiinflammatory effect research. The results show that: 4 kinds of Chinese ephedra processed nux vomica processed by different methods have lower toxicity than the unprocessed nux vomica and the sand-heated product prepared by the pharmacopeia method, and a large number of researches show that the toxicity of the processed nux vomica is reduced, compared with the unprocessed product, the safety of the processed nux vomica is improved, and the clinical application of the nux vomica is expanded. The research results show that the processing purposes of the nux vomica are to reduce toxicity and increase curative effect and application range, and the traditional processing method is low in cost, but the attenuation effect is not ideal, and the operation is complex and time-consuming, so that a novel processing method which has exact curative effect and can greatly reduce the toxicity is needed.

Semen Strychni contains alkaloid, loganin, Doudouchun glycoside, chlorogenic acid, palmitic acid, fatty oil, protein, and polysaccharide. Wherein the total alkaloid accounts for 2 to 5 percent and is the main component. Mainly stryehnine (stryehnine) accounting for about 50% of the total alkali content, and Brucine (Brucine) content accounting for 30% -40%. And small amount of alkaloids such as isostrychnine, isomannine, pseudostrychnine, strychnine oxynitride, loganin, nuxvomicin, colubulin, Nofadrogen, Icaritin, strychnine oxynitride, and 2-hydroxy-3-methoxy brucine. Wherein strychnine and strychnine are the main active ingredients and toxic ingredients, and are also the ingredients which are researched more at present. Literature studies suggest that strychnine and strychnine content is reduced and isostrychnine, isomannine, strychnine oxynitride and strychnine oxynitride content are increased after raw strychnos nux-vomica is subjected to sand-ironing, and it is inferred that strychnine and strychnine are probably converted. Therefore, modern new analysis technology is necessary to deeply research the change and migration of the chemical components of the nux vomica and the processed products thereof, and the material bases of different processing methods can be more comprehensively explained.

Modern pharmacological studies show that nux vomica has the effects of exciting the central nervous system, treating cardiovascular system, resisting inflammation, treating immune system, resisting tumor, relieving pain, relieving cough, eliminating phlegm, relieving asthma, inhibiting bacteria, resisting arrhythmia and the like. The therapeutic dose of nux vomica can improve the excitation process and the inhibition process of cerebral cortex. The toxic dose can make cerebral cortex generate various time phase states, and severe poisoning can cause convulsion of whole body muscle spasticity and gradually leads to death of respiratory muscle spasm contraction asphyxia. Reports have shown that both strychnine and strychnine in extremely large doses prevent the action of cholinesterase on acetylcholine. Nux vomica can also directly damage renal tubular epithelial cells, resulting in acute renal failure and uremia. The researches prove that the nux vomica has definite curative effect but strong toxicity, and the clinical application of the nux vomica is greatly limited, so that a new processing method needs to be developed to achieve the aims of reducing toxicity and improving efficacy.

The applicant and the research team thereof carry out deep research on the attenuation and synergism mechanism of the new nux vomica processing technology, and research results show that a new nux vomica processing technology with two stages is obtained, and a Chinese patent 'a two-stage nux vomica processing method (application number: 201811157918.1)' is applied in 2018, 9, and 30 days; the research aims at improving the chemical structures of strychnine and strychnine which are main active ingredients in nux vomica by a new processing technology so as to achieve the aims of reducing toxicity and improving efficacy. However, the processed nux vomica products still contain a small amount of strychnine and strychnine, and the traditional processed nux vomica decoction pieces also limit the formulation development and clinical application of nux vomica.

Disclosure of Invention

Aiming at the problems, the invention provides a preparation method of a double-order nux vomica processed product extract, which comprises the following steps:

1) weighing semen Strychni, rhizoma Zingiberis recens and semen Phaseoli Radiati according to weight ratio, adding water, and boiling with strong fire for 60 min;

2) taking out semen Strychni, oven drying at 80 deg.C, and scalding with sand to obtain processed semen Strychni;

3) processing semen Strychni, pulverizing, extracting with ethanol solution of pH5 under reflux to obtain extractive solution;

4) recovering ethanol from the extracting solution, adjusting the pH value to 2-3 by using hydrochloric acid, filtering, adjusting the pH value of the filtrate to 9-10 by using ammonia water, evaporating by using a water bath, and drying to constant weight to obtain the double-order nux vomica processed product extract.

In the step 1), the mass ratio of the nux vomica to the ginger to the mung bean to the water is 10:20:1: 150.

And 3) adding 50% ethanol solution with the pH of 5, which is 6 times of the amount of the nux vomica seeds, heating and refluxing for 3 times, extracting for 1 hour each time, and combining ethanol extracting solutions to obtain an extracting solution.

A processed product extract of nux vomica is the processed product extract of nux vomica prepared by any one of the methods.

The active ingredients of the processed product extract of nux vomica are strychnine and alkaloid generated by the reaction of strychnine and phenolic or weakly acidic ingredients in ginger.

The beneficial technical effects of the invention are as follows: 1. the double-stage processed nux vomica product extract provided by the invention overcomes the defect that a small amount of strychnine and strychnine is remained in the nux vomica extract;

2. the volume is reduced, and conditions are created for the formulation development of the processed products of nux vomica.

Drawings

FIG. 1 is a process diagram for establishing a quality loss method;

FIG. 2 is a diagram of the filtration of strychnine and strychnine with mass loss of mother nucleus;

FIG. 3 is a score plot before and after molding;

FIG. 4 shows front and rear opls-da diagrams of molding.

Detailed Description

Example 1 preparation of extracts of processed double-rank Strychnos nux-vomica

With the same amount of nux vomica (100g), the addition amount of ginger, the water addition amount, the ginger decoction time, the nux vomica decoction time and the drying temperature are taken as main investigation factors, detection is carried out according to optimized chromatographic conditions after preparation is finished, and the processing technology is evaluated by taking the contents of strychnine and strychnine as indexes, which are specifically shown in table 1. The optimized processing method finally comprises the following steps: 100g of nux vomica, 200g of ginger and 10g of mung bean are added, 1.5kg of water is added, the mixture is boiled for 60min with strong fire, the mixture is put into the nux vomica to be boiled for 1.5 hours, the mixture is dried in an oven at 80 ℃, and then sand-ironing is carried out to obtain the double-order processed nux vomica product. Adding 50% ethanol solution with pH of 5 and 6 times of semen Strychni, heating and reflux-extracting for 3 times, each time for 1h, mixing the ethanol extracts, filtering, recovering ethanol, adjusting pH to 2-3 with hydrochloric acid, filtering, adjusting pH to 9-10 with ammonia water, evaporating in water bath, and drying to constant weight to obtain the extract of processed product of nux vomica. As can be seen from the liquid phase overlay, brucine and lesioning were not detected in the processed extract of nux vomica.

Example 2 component detection of processed product extract of nux vomica

The method comprises the steps of respectively taking strychnine and strychnine as mother cores, constructing a chemical component database (total 120 compounds) of ginger, presuming the type of reaction between phenolic or weakly acidic components in the ginger and alkaloid, and analyzing TOF data of the processed double-order nux vomica product extract obtained in example 1 by utilizing the mass defect screening function of PeakView software, wherein the TOF data are shown in figures 1 and 2.

Through mass loss, 23 new compounds are identified by taking strychnine as a mother nucleus, and 17 new compounds are identified by taking strychnine as a mother nucleus, which are shown in tables 1 and 2.

TABLE 1 analysis results of novel Compounds

TABLE 2 analysis results of the novel Compounds

Example 3 Activity Studies of processed double-stage Strychnos nux-vomica extract

Continuously gavage the rat for 3d, collecting serum sample after the last administration, establishing a rat toe swelling model, modeling, collecting the serum sample 30min later, centrifuging at 12000r/min ultra high speed, and taking the upper layer serum and a-80 ℃ refrigerator for later use.

Substituting mass spectrum data of samples before and after new process modeling into SIMCA software, establishing an OPLS-DA mathematical model, screening biomarkers through S-PLOT and VIP values, and importing the biomarkers into an established metabolite database to screen differential components by taking the data result of the biomarkers as an index. The results are shown in FIGS. 3 and 4.

ICR mice were divided into 3 groups (blank group, 270mg of crude drug powder of processed product of nux vomica, 11.5mg of extract of processed product of nux vomica (equivalent to 270mg of crude drug of processed product of nux vomica), and each group had 6 mice, male and female. Gavage once a day (0.5ml), gavage for 3 days continuously, after 1h of last administration, exposing the right ear of each group of mice, dripping dimethylbenzene in the middle of the ears of the mice by using a pipette gun except for the normal group, stopping dripping for 5s, dripping dimethylbenzene on the front side and the back side for 30ul, and taking the left ear as a blank control. And (3) after 30min, dislocating the cervical vertebra, cutting off double ears by using surgical scissors, punching down ear pieces at symmetrical positions of left and right ears by using a puncher, weighing by using an electronic balance, recording the weight, and calculating the swelling degree and the swelling rate. The results are shown in Table 3.

TABLE 3 mouse ear swelling test

Claims

1. a preparation method of two-stage strychnine processed product extract, is characterized in that: may further comprise the steps:

1) Weigh strychnids, ginger and mung bean according to weight ratio, add water, boil for 60min on high heat;

2) fish out the strychnids and dry them in an oven at 80°C, then carry out sand scalding to obtain the processed strychnine;

3) concocted strychnine is pulverized, and with the ethanolic solution of pH5, reflux extraction, obtains extract;

4) Recover ethanol from the extract, adjust the pH value to 2-3 with hydrochloric acid, filter, adjust the pH value of the filtrate to 9-10 with ammonia water, evaporate to dryness in a water bath, and dry to constant weight to obtain the double-stage strychnine processed product extract .

2. the preparation method of double-order strychnine processed product extract according to claim 1, is characterized in that: the mass ratio of step 1) described strychnine, ginger, mung bean and water is 10:20:1: 150.

3. the preparation method of two-stage strychnine processed product extract according to claim 1, is characterized in that: step 3) is described as adding 6 times the 50% ethanol solution of strychnine amount, pH=5, Heating and refluxing extraction for 3 times, each time for 1 h, the ethanol extract was combined to obtain the extract.

4. A two-stage strychnine processed product extract is characterized in that: it is the extract of the two-stage strychnine processed product prepared by the method described in any one of claims 1 and 3.

5. double-order strychnine processed product extract according to claim 4 is characterized in that: the active ingredient of double-order strychnine processed product extract is strychnine and strychnine and phenolic or phenolic in ginger. Alkaloids produced by the reaction of weakly acidic components.