WO2019072051A1

WO2019072051A1 - Traditional chinese medicine extract with anti-depressive effect and preparation method and application thereof

Info

Publication number: WO2019072051A1
Application number: PCT/CN2018/103870
Authority: WO
Inventors: 秦雪梅; 赵玉娇; 田俊生; 张潇; 杜冠华
Original assignee: 山西大学
Priority date: 2017-10-12
Filing date: 2018-09-04
Publication date: 2019-04-18
Also published as: CN108653405A; CN107625813A; JP6880236B2; JP2020518596A

Abstract

A traditional Chinese medicine (TCM) extract with an anti-depressive effect and a preparation method and application thereof. The method comprises: collecting, according to a component composition, root of Chinese thorowax, Chinese angelica, largehead atractylodes rhizome stir-fried with wheat bran, white paeony root, peppermint, and licorice root prepared with honey; adding ethanol to extract, filtering, combining filtrates, and concentrating to obtain an ethanol-extracted clear paste; adding water to extract from sediment, filtering, combining filtrates, and concentrating to obtain a water-extracted clear paste; adding ethanol to the water-extracted clear paste, leaving to stand, collecting a supernatant, and concentrating the supernatant into a clear paste; and combining the clear paste and the ethanol-extracted clear paste, mixing and concentrating into a thick paste, microwaving the thick paste to dry under reduced pressure, and pulverizing and sieving to obtain the TCM extract.

Description

Traditional Chinese medicine extract with antidepressant effect, preparation method and application thereof

Technical field

The invention belongs to the field of Chinese medicine extraction, and particularly relates to a traditional Chinese medicine extract with antidepressant effect, a preparation method and application thereof.

Background technique

At present, there are three main treatments for depression: drug therapy, psychotherapy and physical therapy, but still mainly based on chemical drugs. The list of antidepressant drugs listed is limited, the mechanism is single, the efficiency is not high, not more than 70%, and the side effects are large, such as constipation, decreased sexual desire, etc., in the treatment of depression, it also threatens the health of other functions of the patient's body. Clinically, when Western medicine is used to treat depression, comprehensive consideration should be given to the treatment efficiency, mechanism of action, dose selection, side effects, drug interaction and other factors, and for some complex depressions such as bipolar polyphasic depression, one or more Symptoms of disease and depression are not ideally improved.

It is precisely because of the defects of existing western medicines and the urgency of market demand that scholars at home and abroad have turned their attention to Chinese herbal extracts. How to develop from Chinese herbal medicine on the basis of clear anti-depressant efficacy, or have its own unique pharmacodynamic characteristics, or have a new anti-depressant mechanism, or for specific conditions, or have new indications, or have relatively few side effects New antidepressant drugs with multi-target, multi-effect and other features are the major trends in the development of antidepressant drugs in the future.

Xiaoyao San is a classic Chinese medicine prescription for soothing the liver and relieving depression. In the previous period, the two authorized patents of this research group (201010110283.7 and 201110084715.6) extracted with ethyl acetate and petroleum ether, indicating that Xiaoyao San has a clear anti-depression effect, but on the one hand, industrial production has many safety hazards, and the process controllability is poor; Xiaoyao Sanyuan prescription has more medicinal materials, and the dosage is relatively large, and the quality is not easy to control. Therefore, based on the research on Xiaoyao San, the present invention prepares a traditional Chinese medicine extract by using the alcoholic water extraction method which is simpler in composition and more suitable for the pilot production, and studies the antidepressant effect characteristics.

Summary of the invention

The object of the present invention is to provide a traditional Chinese medicine extract with simple composition, scientific and reasonable technology, and a plurality of medicinal properties with significant improvement in depression, and a preparation process and application thereof.

The invention provides a traditional Chinese medicine extract with antidepressant effect, which is characterized by a distribution ratio and a preparation method of the traditional Chinese medicine group:

The traditional Chinese medicine component is in terms of mass parts: Bupleurum 20-60, Angelica 20-60, bran-frying atractylodes 20-60, white peony 20-60, mint 8-20, licorice licorice 8-20;

The preparation method comprises the following steps: 1) adding 6-10 times weight of 60-90% ethanol to the component medicine, extracting twice, each time 1-3 h, filtering, combining the filtrate, and concentrating to obtain alcohol extract 2) The dregs are added with 6-10 times the weight of water for 2 times, each time 1-3h, filtered, the filtrate is combined, concentrated to obtain a water extracting paste, and ethanol is added to the water extracting paste to make the alcohol content Up to 60-80%, let stand for 24h, take the supernatant, concentrate to clear the paste; 3) combine the clear paste with the alcohol extract paste, mix and concentrate to thick paste, dry under microwave pressure, smash through 80 mesh sieve , that is, the Chinese medicine extract.

The components are preferably: Bupleurum 30, Angelica 30, Brassica chinensis 30, Angelica 30, Peppermint 10, and Zhigancao 15 in parts by mass.

The weight of the ethanol added in the step 1) is 8 times the weight of the medicinal material, the ethanol concentration is 75%, and the extraction time is 2 hours.

The weight of the added water in the step 2) is 8 times the weight of the medicinal material, and the extraction time is 2 hours.

Step 3) The water content of the water extracting paste after adding ethanol is 70%.

The density of the above alcohol extracting paste, water extracting paste and clearing paste is controlled to be 1.08 to 1.10, and the density of the thick paste is controlled to be 1.24 to 1.26.

The traditional Chinese medicine extract as described above has been experimentally proven to have significant antidepressant effects, which can be applied in the preparation of a medicament for treating depression.

The invention provides a traditional Chinese medicine granule comprising the traditional Chinese medicine extract and the auxiliary material and the like as described above.

The invention has the following advantages:

1. The invention provides that the prescription is simpler, the dosage is less, the quality is more controllable, the pharmacological activity reaches or exceeds the listed antidepressant western medicine venlafaxine, the antidepressant traditional Chinese medicine Shugan Jieyu and Xiaoyao San.

2, the use of more reasonable and safe process, easier to pilot industrial production of alcohol water extraction method, the maximum extent, more comprehensive extraction of active ingredients, remove impurities, reduce the rate of extraction to reduce the dosage of patients.

3. Compared with the commonly used antidepressant western medicine venlafaxine, the antidepressant effect is equivalent, and: 1 no venlafaxine-induced constipation, decreased gastrointestinal motility and decreased sexual desire; 2 improved CUMS-depressed rats Learning memory decline and anxiety.

DRAWINGS

Figure 1 is a flow chart of the preparation process of the traditional Chinese medicine extract of the present invention

Detailed ways

The following specific examples are set forth to illustrate the invention, but are not intended to limit the scope of the invention.

Example 1

Weigh 30 parts of Bupleurum, 30 parts of Angelica, 30 parts of bran atractylodes, 30 parts of white peony, 10 parts of menthol, 15 parts of licorice, and add 8 times of 75% ethanol for 2 times, 2h each time. After filtration, the filtrate was combined and concentrated to an alcohol extracting paste having a relative density of 1.09; the dregs were added to 8 times by weight of water for 2 times, each time for 2 hours, filtered, and the filtrate was concentrated to a water having a relative density of 1.09. Clear the paste; add ethanol to the water extract paste to make the alcohol content up to 70%, let stand for 24h (4 ° C), take the supernatant to concentrate to a relative density of 1.09 clear paste, and mix with the alcohol extract paste, mix After concentration, it is concentrated to a thick paste having a relative density of 1.25, and dried under reduced pressure in a microwave to obtain a dry extract, which is pulverized and passed through a sieve of 80 mesh.

Example 2

Weigh 30 parts of Bupleurum, 30 parts of Angelica, 20 parts of braised atractylodes, 20 parts of white peony, 10 parts of menthol, 15 parts of licorice, and add 2 times of 60% ethanol for 2 times, 2h each time. After filtration, the filtrate was combined and concentrated to an alcohol extract with a relative density of 1.08; the dregs were added to 8 times by weight of water for 2 times, each time for 2 h, filtered, and the filtrate was concentrated to a water density of 1.08. Clear the paste; add ethanol to the water extract paste to make the alcohol content up to 60%, let stand for 24h (4 ° C), take the supernatant to concentrate to a relative density of 1.08, and mix with the alcohol extract paste, mix After concentration, it was concentrated to a thick paste having a relative density of 1.24, and dried under reduced pressure in a microwave to obtain a dry extract, which was pulverized and passed through a sieve of 80 mesh.

Example 3

Weigh 30 parts of Bupleurum, 30 parts of Angelica, 30 parts of bran atractylodes, 20 parts of white peony, 10 parts of menthol, 15 parts of licorice, and add 8 times of 90% ethanol for 2 times, 2h each time. , filtered, the filtrate was combined, concentrated to a relative density of 1.10 alcohol extract paste; the drug residue was added to 8 times the weight of water for 2 times, each time 2h, filtered, the filtrate was combined, concentrated to a relative density of 1.10 water extract Clear the paste; add ethanol to the water extract paste to make the alcohol content up to 80%, let stand for 24h (4 ° C), take the supernatant to concentrate to a relative density of 1.10 clear paste, and mix with the alcohol extract paste, mix After concentration, it was concentrated to a thick paste having a relative density of 1.26, and dried under reduced pressure in a microwave to obtain a dry extract, which was pulverized and passed through a sieve of 80 mesh.

Example 4 Antidepressant efficacy test

4.1 experimental animals

SD adult male healthy rat (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number: SCXK (Beijing) 2014-0012)

4.2 Experimental drugs and herbs

Venlafaxine hydrochloride (Chengdu Kanghong Pharmaceutical Group Co., Ltd., specification: 25mg/granule, batch number: national medicine standard word H20070269)

Shugan Jieyu Capsule (Chengdu Kanghong Pharmaceutical Group Co., Ltd., Specification: 0.36g/granule, batch number: National Pharmaceutical Standard Z20080580)

The traditional Chinese medicine extract prepared in Example 1, wherein Bupleurum, Angelica, Bran White Atractylodes, Radix Paeoniae Alba, Radix Glycyrrhizae and Peppermint were purchased from Shanxi Huayang Pharmaceutical Co., Ltd., and Professor Qin Xuemei, Director of Modern Research Center of Traditional Chinese Medicine of Shanxi University The identification is genuine.

4.3 Animal grouping and administration

Rats were adaptively reared for 1 week, during which syrup training was carried out. Rats were randomly divided into 7 groups according to body weight, with 10 rats in each group.

1 blank control group (K)

2 model group control group (M)

3 venlafaxine group (35mg/kg) (Y)

4 Shugan Jieyu group (150mg/kg)(S)

5 Chinese medicine extract high dose group (16.6g crude drug / kg) (G)

6 Chinese medicine extract dose group (8.3g crude drug / kg) (Z)

7 Chinese medicine extract low dose group (4.2g crude drug / kg) (D)

All the drugs were dissolved in distilled water, and each group of animals was intragastrically administered once a day (1 ml/100 g) according to the body weight.

4.4 Model establishment

Stimuli include fasting, water ban, 4°C ice water swimming, 50°C thermal stimulation, tail trapping, intermittent foot shock, restraint, ultrasound stimulation, and day and night reversal. One stimulus was randomly given daily, and each stimulus was used 2-3 times cumulatively. Except for the blank control group, the other groups were housed in a single cage and the modeling procedure was carried out. The modeling time was 28 days. After the start of modeling, the drug was administered by intragastric administration. The drug group was given the corresponding concentration of the drug solution, and the model group and the blank control group were given an equal volume of distilled water.

4.5 detection indicators

Body weight: Rat body weight was weighed on the 0th, 7th, 14th, 21st, and 28th day of the experiment.

Sugar water preference rate: After the end of modeling, the rat 1% sugar water consumption was measured, and the sugar water preference rate was calculated.

Open field experiment: Rats were placed in the open field box for 5 min, and the number of penetrating and erecting times of the rats within 4 min after recording were recorded.

4.6 Experimental results

The effects of Chinese herbal extracts on body weight, sucrose preference rate and open field test of CUMS rats are shown in Tables 1, 2 and 3: Chinese herbal extracts can improve the growth of rat body weight caused by CUMS modeling stress. In the field experiment, the decrease of activity and the preference for sucrose sweetness indicate that the extract of Chinese medicine has a good antidepressant effect, and its pharmacological activity meets or exceeds the listed antidepressant western medicine venlafaxine and antidepressant Chinese medicine Shugan solution. Yu.

Table 1 Effect of Chinese Herb Extract on Body Weight of CUMS Rats

Compared with the blank group, #p<0.05, ##p<0.01, compared with the model group, *p<0.05, **p<0.01

Table 2 Effect of Chinese herbal extracts on saccharide water preference rate in CUMS rats

Table 3 Effect of Chinese Herb Extract on the Field Experiment of CUMS Rats

Example 5 Antidepressant efficacy--Improvement of gastrointestinal function in CUMS depressed rats

5.1 experimental animals

Same as Embodiment 4.

5.2 Experimental drugs and herbs

Same as Embodiment 4.

5.3 Animal grouping and administration

Same as Embodiment 4.

5.4 Model establishment

Same as Embodiment 4.

5.5 Test indicators

Feces: After the modeling was completed, the rats in each group were placed in metabolic cages, and the feces of the rats were collected in a plastic beaker for 12 hours. The amount of feces was counted, weighed (wet weight), and dried in an oven (70 ° C, 3 h). Weighing (dry weight). Calculate the moisture content of the stool = (wet weight dry weight) / wet weight × 100%.

Intestinal propulsion and gastric emptying experiments: The day before the death, fasting can not help but water for 12h. On the next day, 30 minutes before the rats were sacrificed, the self-made semi-solid nutrient paste was administered according to the dose of 1ml/100g. After 30 minutes, the rats were sacrificed, and the whole stomach was quickly dissected, and the gastric cardia, gastric pylorus and ileocecal part were ligated, and dried with filter paper. Regain total stomach weight (g) and small intestine. The surgical scissors were cut along the large curvature of the stomach wall, and the contents of the stomach were washed with distilled water, and the filter paper was dried and weighed to obtain the net weight (g) of the stomach.

Gastric residual rate = (stomach weight - stomach net weight) / homemade semi-solid nutrient paste weight × 100%

Small intestine propulsion rate = length of small intestine / distance of self-made semi-solid paste propulsion × 100%

5.6 Experimental results

Table 4 shows that CUMS stress increases the number of bowel movements in rats and decreases the water content of feces. Compared with venlafaxine, this phenomenon can not be improved and negative regulation is observed. After the administration of traditional Chinese medicine extract, the abnormality of the above indicators can be improved, indicating that the Chinese herbal extract improves the depression-like symptoms of CUMS rats without causing fecal abnormalities. side effect. Tables 5 and 6 show that modeling stress slows the intestinal peristalsis and gastric emptying function in rats, and venlafaxine has a decreasing trend in the small intestine propulsion rate and intragastric meal rate compared with the model group, suggesting that venlafaxine It may have the effect of reducing gastrointestinal function. Chinese herbal extracts can improve gastrointestinal function in rats while treating depression.

Table 4 Effect of Chinese Herb Extract on Feces of CUMS Rats

Table 5 Effect of Chinese Herb Extract on Small Intestinal Propulsion Rate in CUMS Rats

Table 6 Effect of Chinese Herb Extract on Gastric Emptying Rate in CUMS Rats

Example 6 Antidepressant efficacy characteristics - improving sexual function in CUMS depressed rats

6.1 experimental animals

Same as Embodiment 4.

6.2 Experimental drugs and herbs

Same as Embodiment 4.

6.3 Animal grouping and administration

Same as Embodiment 4.

6.4 Model establishment

Same as Embodiment 4.

6.5 test indicators

48 h before the experiment, female mice were injected subcutaneously with estradiol 10 μg (dissolved in 0.1 ml safflower oil); 2 h before the experiment, 10 μg of progesterone was injected subcutaneously. The CUMS male and the estrus female were caged, and at 7-9 pm, the males' sexual behavior was observed within 30 minutes, and the crawling latency and the number of crawling backs were recorded.

6.6 Experimental results

The crawling behavior of male rats in mating experiments can reflect the size of their sexual desire to a certain extent. Table 7 shows that CUMS stress prolongs the crawler latency of the rats and reduces the number of crawling backs. To some extent, the arousal of rat sexual desire is affected by the modeling stress. Compared with venlafaxine, this indicator is negatively regulated. After administration of traditional Chinese medicine extract, the abnormality of the above indicators can be improved, indicating that the Chinese herbal extract improves the depression-like symptoms of CUMS rats without causing side effects of libido reduction.

Table 7 Effect of Chinese Herb Extract on Sexual Desire in CUMS Rats

Example 7 Antidepressant efficacy--Improvement of learning and memory decline and anxiety in CUMS-depressed rats

7.1 experimental animals

Same as Embodiment 4.

7.2 Experimental drugs and herbs

Same as Embodiment 4.

7.3 Animal grouping and administration

Same as Embodiment 4.

7.4 Model establishment

Same as Embodiment 4.

7.5 Test indicators

After the modeling, the Morris water maze and the elevated cross maze experiment were carried out. The spatial navigation latency and the number of crossing platforms in the water maze were selected as the investigation of the cognitive function of CUMS rats. The number of open arms in the elevated cross maze was selected. Proportion and ratio of time to open arm were used as an examination of anxiety in CUMS rats.

7.6 Experimental results

Table 8 shows that CUMS-depressed rats have a reduced number of crossing platforms and target quadrants during the spatial exploration period, suggesting that CUMS-depressed rats are associated with cognitive impairment. Compared with venlafaxine, this phenomenon can be adjusted, and the Chinese herbal extract can improve this symptom after administration.

Table 8 Effect of Chinese Herb Extract on Learning and Memory in CUMS Rats

The proportion of CUMS-depressed rats entering the open time in the elevated plus maze and the proportion of entering the opening wall decreased, suggesting that CUMS depressed rats were associated with anxiety. Both venlafaxine and Chinese herbal extracts can improve this symptom after administration.

Table 9 Effect of Chinese Herb Extract on Anxiety of CUMS Rats

Example 8 Sugar-free granules

The traditional Chinese medicine extract powder and the pregelatinized starch are mixed in a ratio of 5:2, and are granulated by dry granulation to prepare granules. The parameters of the dry granulator are adjusted to: extrusion speed 6.0 rpm; feed speed 7.0 rpm; The pellet speed was 10.0 rpm; the side seal pressure was 13 MPa; the extrusion pressure was 27 MPa. The mesh used in the granulation was 10 mesh, and the whole sieve was 40 mesh. The granules are granulated and packaged by the composite film to obtain the finished product.

Example 9 Sugar-containing granules

Chinese herbal extract powder is mixed with sucrose and dextrin in a ratio of 2:0.5:0.5, using 75% ethanol as a wetting agent, swaying granulator, passing through 10 mesh stainless steel mesh, granulating, drying, passing 40 mesh sieve The net is granulated and packaged in a composite film.

Claims

An extract of traditional Chinese medicine having antidepressant effect, characterized by a distribution ratio and a preparation method of a traditional Chinese medicine group;

The traditional Chinese medicine component is in terms of mass parts: Bupleurum 20-60, Angelica 20-60, bran-frying atractylodes 20-60, white peony 20-60, mint 8-20, licorice licorice 8-20;

The preparation method comprises the following steps: 1) adding 6-10 times weight of 60-90% ethanol to the component medicine, extracting twice, each time 1-3 h, filtering, combining the filtrate, and concentrating to obtain alcohol extract 2) The dregs are added with 6-10 times the weight of water for 2 times, each time 1-3h, filtered, the filtrate is combined, concentrated to obtain a water extracting paste, and ethanol is added to the water extracting paste to make the alcohol content Up to 60-80%, let stand for 24h, take the supernatant, concentrate to clear the paste; 3) combine the clear paste with the alcohol extract paste, mix and concentrate to thick paste, dry under microwave pressure, smash through 80 mesh sieve , that is, the Chinese medicine extract.
The traditional Chinese medicine extract according to claim 1, wherein said components are in parts by mass: Bupleurum 30, Angelica 30, Brassica chinensis 30, Angelica 30, Peppermint 10, and Zhigancao.
The traditional Chinese medicine extract according to claim 1, wherein the weight of the ethanol added in the step 1) is 8 times the weight of the medicinal material, the ethanol concentration is 75%, and the extraction time is 2 hours.
The traditional Chinese medicine extract according to claim 1, wherein the weight of the added water in the step 2) is 8 times the weight of the medicinal material, and the extraction time is 2 hours.
The traditional Chinese medicine extract according to claim 1, characterized in that the alcohol content of the water extracting paste in step 3) after adding ethanol is 70%.
The traditional Chinese medicine extract according to claim 1, wherein the density of the alcohol extracting paste, the water extracting paste and the clearing paste is controlled to be 1.08 to 1.10, and the density of the thick paste is controlled to be 1.24 to 1.26.
The use of the traditional Chinese medicine extract according to claims 1 to 6 for the preparation of a medicament for treating depression.
A traditional Chinese medicine granule comprising the traditional Chinese medicine extract according to any one of claims 1 to 6.