WO2006097043A1

WO2006097043A1 - Bacopa monnieri extract and process for preparation and use thereof

Info

Publication number: WO2006097043A1
Application number: PCT/CN2006/000394
Authority: WO
Inventors: Mingfu Yang; Yunxiu Huang; Yuping Lei
Original assignee: Chengdu Wagott Pharmaceutical Co., Ltd.
Priority date: 2005-03-15
Filing date: 2006-03-14
Publication date: 2006-09-21
Also published as: CN1833692A; CN1833692B

Abstract

The present invention provides Bacopa monnieri extract, wherein total saponin of B. monnieri comprising 10% to 80% by weight of the extract, in terms of bacoside A. The present invention also provides pharmaceutical compositions comprising B. monnieri extract and process for preparation and use thereof. The present B. monnieri extract of various amount of bacoside A present is useful in various indications, with extensive clinical usage, reduced cost, enhanced controllability, and stable quality.

Description

Pseudomonas sinensis extract and preparation method and use thereof

The present invention relates to a shamrock extract, and more particularly to an extract of a natural medicinal material, Purslane, which comprises a preparation method and use of the extract, and belongs to the field of medicine.

Background technique

The fake purslane is the whole grass of Bacopa monninera, a traditional Indian herb. It has the effects of clearing away heat and detoxifying, reducing inflammation and reducing swelling, treating red eyes and dysentery, redness and swelling of the eyes, redness and swelling of the skin, etc. It is a well-known antipsychotic and memory-enhancing drug in India.

At present, reports on the false purslane are: Yang Xiaofeng, Song Chunqing, an artifact in the study of natural products, Journal of Pharmacy, ₁₉ 99, 34 (12), which discloses that the alcohol extract of the Indian herbal purslane can be obvious. Improve the cognitive ability of mice and improve the memory of mice. The active ingredient is a mixture of various saponins.... This paper only summarizes the efficacy of extracts of P. oleracea L., and the efficacy test of no active ingredients. There is no detailed extract preparation process; the analgesic effect of new triterpenoid bacosine in P. oleracea, Chinese herbal medicine, 1998, 5, discloses the analgesic effect of triterpenoids in P. oleracea. Gingival sputum is an "adaptogen" with two-way regulation and anti-stress response, which can help regulate the body's ability to resist environmental stress and guide the return to normal health. D. Rai et al. The effects of extracts on acute stimulation (AS) and chronic stimulation (CS) model rats showed that: extracts containing pseudo-dental saponins as the main component were taken 45 minutes earlier. Will significantly improve a series of reactions caused by AS, such as gastric ulcer index, adrenal gland weight, elevated blood glucose levels, aspartate aminotransferase activity, alanine transaminase activity and creatine kinase activity, and spleen weight Long-term stimulation experiments show that: when the low dose of P. oleracea extract will reduce the index of gastric ulcer and the activity of aspartate aminotransferase, and tend to 1 έ. High dose will make the gastric ulcer index, adrenal gland The weight decreases, and the aspartate aminotransferase and creatine kinase activities decrease and tend to be normal. Therefore, the extract of the purslane has a soothing effect, is an environmental hormone, and adapts to the original. Portulaca oleracea is a calcium blocker that has an antispasmodic effect on smooth muscle. Some people also mix the extract of P. oleracea and other ingredients into various dosage forms to improve menopausal women's estrogen reduction. Discomfort, also used in beauty and skin care.

The existing literature is mainly on the identification of its chemical constituents, especially the triterpenoid saponin structure. The extraction process of the saponin is as follows: The dried saponin is extracted with water or alcohol, separated by a solvent method or separated and purified by silica gel column chromatography. Represented by the following two patents - the first one is: WO2004060267-A2 The extraction and separation methods are as follows: water or organic solvent extraction, drying, removal of fatty substances in a non-polar solvent, extraction of residues with at least one organic solvent, extraction with water, drying of the organic layer under vacuum, separation by silica gel column chromatography, water or organic The solvent is eluted to obtain a product having a saponin A+B content of 80-100%, and A and hydrazine are separated by chromatography.

The second part is: WO2004054599-A1, extracting saponin A+B content of 20~30% extract.

Decondensation of n-hexane, drying of filter residue, extraction of acetone (removal of non-BACOSIDE components and pigment substances), extraction of methanol residue, concentration of extract, addition of concentrate to acetone, filtration, filtration of water, filtration of n-butanol, concentration The n-butanol solution was dried, the dried product was dissolved in water, and a non-toxic stabilizer (such as e-cyclodextrin) was added, stirred for 1.5 hours, and spray dried.

The above two patents, the first one is mainly to extract a single component, a higher purity product, which can be used as a standard product; the second preparation of saponin A+B content is 20~30°/. Extract. Both patents use more organic solvents.

In addition, the extraction method of P. oleracea, the literature reports: First, use water extraction, discard the aqueous extract, and then extract twice with 90% ethanol. Extract with acetone. The BACOSIDES content in BACOPA is 50-60% based on BACOSIDE A. (Adaptogenic effect of Bacopa monniera (Brahmi), D. Rai et al. I Pharmaco Iogy, Biochemistry and Behavior 75 (2003) 823-830.) The extraction process is separated and purified by acetone, using an organic solvent harmful to human body such as acetone. It is unfavorable for large industrial production operations and has high cost. Summary of the invention

The technical solution of the present invention is to provide a extract of P. oleracea, and another technical solution of the present invention provides a preparation method and use of the extract.

The present invention provides a extract of P. oleracea, wherein the total percentage of total saponins of P. oleracea is from 10% to 80% by weight of Bacoside A.

BACODSIDE A consists of the following components:

Bacoside A ₃ , bacopaside II, bacopaside X, bacopasaponin C, the structure is as follows: bacoside A3: The aglycone is jujube saponin, the sugar chain is:

3-0-β-D-glucopyranosyl-(1~► 3) - 0- [aL-furan arabinosyl-(1 -^ 2) ]-0- (β-D-glucopyranosyl)

Bacopaside II: The aglycon is: pseudojujube saponin, the sugar chain is:

3 - 0- a- L -furan arabinosyl-(I 2) - [ 0 - D -glucopyranosyl-(1 3) ] - β -D-pyranosyl

Bacopaside X: aglycon is: jujube saponin, the sugar chain is: 3-0- a-L-furan arabinosyl-(1 _► 2) - [β-D-glucopyranosyl-(1·► 3) - ] - α - L-pyranose

Bacopasaponin C:

The aglycon is: pseudojujube saponin, the sugar chain is: 3- O-β-D-glucopyranosyl-(1 -[ aL-furan arabinyl-(1- 2) ] - a- L pyran Arabinosyl

Bacopaside X

Bacopasaponin C

Further, the total percentage of total saponins of P. oleracea is in the range of Bacoside A, which accounts for 10% to 40% of the extract. Preferably, the total percentage of total saponins of P. oleracea is from 10% to 30% by weight of Bacoside A. Further preferably, the total percentage of total saponins of P. oleracea is in the range of Bacoside A, which is 20% ± 0.04 of the extract.

Or, further, the total percentage of total saponins of P. oleracea is in the range of Bacoside A, which accounts for 40% to 70% of the extract. 01。 The total weight of the total saponin of the saponin is determined by Bacoside A, accounting for 50% ± 0.1 of the extract.

Among them, the false purslane is derived from the whole grass of the scrophulariaceae plant Bacopa monnieri L.

The extract of the scutellaria chinensis is extracted by the following method: taking the whole grass of the dried scorpionfish Bacopa monnieri L., extracting 40%~80% ethanol, and extracting the extract directly and drying to obtain the extract of the scutellaria Or after concentrating the extract, the macroporous adsorption resin is adsorbed, and then eluted with 10% to 95% ethanol, and the eluate is collected and dried to obtain.

The invention also provides a method for preparing the extract of the purslane, which comprises the following steps: a, weighing the whole grass of Bacopa monnieri L., adding 40%~80% ethanol extraction, and extracting the extract, Concentrated liquid;

b. The concentrated extract is purified and purified by a macroporous adsorption resin method, and 10-95% ethanol is used as an eluent, and the eluate is collected and dried to obtain.

Among them, the ethanol elution described in step b is: eluted with 60-95% ethanol. Further, the ethanol elution described in step b is: first eluted with 10% to 60% ethanol, and then with 60% to 9% ethanol.

Among them, elution can be carried out in the following two ways: (1) directly eluted with 60 to 95% ethanol. (2) First elute with 10%~60% ethanol, and then elute with 60%~95% ethanol when the color of the eluent is obviously lighter. The present invention also provides the use of the extract of Pseudomonas sinensis in the preparation of an antidepressant and puzzled drug.

The present invention also provides a pharmaceutical composition which is prepared by adding an effective amount of a extract of P. oleracea as a raw material, and adding a pharmaceutically acceptable excipient or an excipient.

Wherein the preparation is an oral preparation or an injection.

The invention also provides the use of the pharmaceutical composition for the preparation of a medicament for antidepressant. Wherein, the weight percentage of the total saponins of Pseudomonas sinensis in the drug containing Baccoon A is 10% to 80% of the extract. Preferably, the pseudo-dentate width extract of the medicament contains the total content of the total saponins of the purslane in a Bacoside A ratio of 10 to 40%. Further preferably, the drug has a weight percentage of the total saponin of the purslane, which is 20% ± 0.04 in terms of Bacoside A.

The invention also provides the use of the pharmaceutical composition for the preparation of a puzzle drug. Wherein, the weight percentage of the total saponins of Pseudomonas sinensis in the drug is Bacoside A, which accounts for 10% to 80% of the extract. Preferably, the drug contains 0.1% by weight of the total saponins of P. oleracea extract in terms of Bacoside A, and is 40-70% by weight. Further, preferably, the drug has a weight percentage of the total saponin of the scutellaria sinensis in the medicinal saponin of the scutellaria saponin, which is 50% ± 0.1 in terms of Bacoside A.

Wherein the preparation is an oral preparation or an injection. A medically acceptable variety of preparations consisting of extracts of P. oleracea or extract of P. oleracea and pharmaceutical excipients. Such as tablets, capsules, soft capsules, granules, injections, and the like. The auxiliary materials used in the present invention are excipients commonly used in pharmaceutical engineering such as solid preparations: dextrin, starch, starch derivatives, carboxymethyl cellulose, sucrose, calcium sulfate, magnesium stearate, lactose, micro-fine cellulose , glucose, sorbitol, mannitol, etc. Excipients used in liquid preparations such as: water, ethanol, glycerin, propylene glycol, polyethylene glycol, and the like.

The extract of the purslane of the present invention may be a low content of 10 to 30%, or a high content of 60 to 80%, and different specifications of the extract of the purslane have different effects. In the production process of raw materials, it is not necessary to degrease the product, the resin is used for adsorption and impurity removal, the processing amount in the production process is large, and it can be reused through activation, which saves cost, and the organic solvent involved only ethanol, reducing organic solvent Damage to operators and production pipelines. Compared with the extract prepared by other methods, the extract of the purslane extract of the present invention has low cost, remarkable drug effect, strong controllability and stable quality.

It is apparent that various other modifications, substitutions and changes can be made in the form of the above-described basic technical concept of the present invention without departing from the spirit and scope of the invention.

The above content of the present invention will be further described in detail below by way of specific embodiments of the embodiments. Bright. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. Any technique implemented based on the above description of the present invention is within the scope of the present invention.

DRAWINGS

Figure 1 Mouse platform test - Mean latency map of each group of mice (24-hour memory retention) (S) Figure 2 Mouse platform test ^ Mean mouse latent period (long-term memory retention) mean point map S) Figure 3 Antidepressant Screening - Forced Mouse Swimming Test (Day 9) Mouse Immobility Time Mean (S) Figure 4 Antidepressant Screening - Forced Mouse Swimming Test (Day 15) Small Mouse immobility time average point map (S)

Example 1 Preparation of Extract of Pseudomonas avenae of the present invention

350g of pseudo-horse wide dry whole grass, extracted with 80% ethanol reflux three times, the amount is 10, 8, 6 times, each extraction for 1 hour, the three extracts are filtered, concentrated and dried to obtain 60.6g of product, Bacoside A The weight percentage is 10.5%.

Example 2 Preparation of the pseudo-tooth width extract of the present invention

200 fake dentate wide dry whole grass, extracted with 40% ethanol reflux three times, the amount is 10, 8, 6 times, each extraction for 1 hour, the three extracts are filtered and concentrated to no alcohol, macroporous adsorption resin adsorption, 7J was washed until it was colorless, first eluted with 10% ethanol, and then eluted with 95% ethanol. The 95% ethanol eluate was concentrated and dried to obtain 3.4 g of a product. The weight percentage of Bacoside A was 21.11%.

Example 3 Preparation of the extract of the scutellaria

60kg whole grass material, extracted twice with 70% ethanol reflux, the dosage is 10, 10 times, each extraction for 1 hour, filtering and combining the filtrate, the filtrate is concentrated on the column, washed with water until colorless, washed with 95% ethanol 120L Degassed, spray dried eluate to obtain 200 _{g of} product, Bacoside A weight percentage: 31.1%.

Example 4 Preparation of the extract of Pseudomonas avenae of the present invention

350g of dried purslane dried whole grass, 80% ethanol reflux three times, the dosage is: 8, 8, 6 times, each extraction for 1.5 hours, the three extracts are filtered and concentrated to 600ml, macroporous adsorption resin adsorption, first Wash with water until colorless, elute with 600 ml of 10% ethanol then elute with 1000 ml of 95% ethanol. The 95% ethanol eluate was concentrated and dried to obtain 6 g of dry powder. Bacoside A has a weight percentage of 43.24%.

Example 5 Preparation of the pseudo-tooth width extract of the present invention

350 _{g of} purslane dried whole grass, 80% ethanol reflux three times, the dosage is: 8, 8, 6 times, each extraction for 1.5 hours, the three extracts are filtered and concentrated to 600ml, first with 40% 600ml ethanol Elution, elution with 1000 ml of 95% ethanol, concentration and drying of 95% ethanol activation solution, to obtain 5.6 _{g of} product. Bacoside A's weight Percentage by weight: 50.37 %.

Example 6 Preparation of the extract of the scutellaria

350g of pseudo-purslane dried whole grass, extracted with 80% ethanol reflux three times, the amount is 10, 8, 6 times, each extraction for 1 hour, the three extracts are filtered and concentrated to no alcohol, macroporous adsorption resin adsorption, Wash with water until colorless, first elute with 30% ethanol, elute with 95% ethanol, concentrate 95% ethanol eluate, and dry. The product has a yield of 5.8 g and Bacoside A has a weight percentage of 60.8%.

Example 7 Preparation of the extract of the scutellaria

350 _{g of} scutellariae dried whole grass, three times with 70% ethanol reflux, the amount is 10, 8, 6 times, each extraction for 1 hour, the three extracts are filtered and concentrated to no alcohol, macroporous adsorption resin Adsorption, washing to colorless, eluting with 30% ethanol, eluting with 95% ethanol, concentrating 95% ethanol eluate, and drying. The product was obtained in an amount of 5 g, and Bacoside A had a weight percentage of 72.5%.

Example 8 Preparation of Extract of Pseudomonas avenlus of the present invention

350 Falcon purslane whole grass, extracted with 70% ethanol reflux three times, the amount is 10, 8, 6 times, each extraction for 1 hour, the three extracts are filtered and concentrated to no alcohol, macroporous adsorption resin adsorption, water washing To colorless, first elute with 30% ethanol, elute with 90% ethanol, concentrate 90% ethanol eluate, and dry. The product was 4.1 g, and Bacoside A had a weight percentage of 79.1%.

Example 9 Method for detecting extract of P. oleracea

The quality control methods of the extracts prepared in Examples 1-8 are as follows:

Instruments and reagents:

HP 1100 quaternary pump; HP 1100 column incubator; HP 1100 detector; reference purity ≥97%; instrument water for re-distilled water; experimental water for heavy distilled water; acetonitrile for chromatographic purity; other reagents are of analytical grade.

Chromatographic conditions:

Column: Inertsil ODS 150 X 4.6mm 5um

Mobile phase:

Column temperature: 30 ° C; flow rate: 1.0 ml / min; detection wavelength: 205 nm; injection amount: 20 ul; sensitivity: 0.01 AUFS. Solution preparation:

1. Preparation of standard solution

The reference substance of Bacoside A ₃ , bacopaside II, bacopaside X, bacopasaponin C dried to a constant weight by phosphorus pentoxide was accurately weighed, and methanol was added to make a mixed solution containing 0.15 mg per 1 ml, which was used as a reference solution.

2. Preparation of test solution Weigh accurately two samples of 100mg in two 50ml volumetric flasks, add about 40ml of analytical methanol, sonicate in ultrasonic, let cool to room temperature, dilute to volume, shake well and use 0.45um microporous membrane Filter, that is.

Sample determination:

The test sample was injected into the liquid chromatograph, and the peak area was recorded and calculated according to the external standard method.

The content of bacoside A in this product is based on the total amount of Bacoside A ₃ , bacopaside II, bacopaside X, and bacopasaponin C.

Example 10 Preparation of the pharmaceutical composition of the present invention

The following materials were weighed by weight: The extract of Pseudomonas cuspidatum of the present invention prepared in Example 1 was 60%, and 20% of fine cellulose, stevioside 5°/. , starch 10%, magnesium stearate 0.8%, hydroxypropyl cellulose 4.2%, fully mixed and granulated, packaged in the required dosage to obtain granules.

The granules obtained according to the examples were dispensed into the empty plastic shell at the required dosage to obtain a capsule.

The granules obtained according to the examples were directly tableted and film-coated to obtain tablets.

The above extract 30 _g , with water for injection, was completely dissolved under aseptic conditions, respectively, and passed through a glass filter G3, G6, potted in an ampoule, and sterilized at 100 ° C for 30 min to obtain 1000 pieces.

The pharmaceutically acceptable preparations of different specifications can be prepared by using the extracts of different specifications as described.

The beneficial effects of the present invention are demonstrated by pharmacodynamic tests below.

Test materials and methods -

1. Test sample - The extract of P. oleracea prepared according to the method described in Examples 1-8, calculated as Bacoside A:

A: 21.11% B: 50.37% C: 72.50%

2. Dosage setting and preparation of the test article of the present invention (the test sample is prepared according to the method of the embodiment: corresponding samples are respectively examples 2, 5, 7)

Table 1 Dose setting and preparation of test samples

Dose concentration

Group (mg/kg) ig. (mg/nil)

A low dose group 30 1. 5

Medium dose group 60 3. 0

High dose group 120 6. 0

B Low dose group 30 1. 5

Medium dose group 60 3. 0

High dose group 120 6. 0

C Low dose group 30 1. 5

Medium dose group 60 3. 0

High-dose group 120 6. 0 The above test samples are sealed and stored at room temperature. 3. Positive control and preparation

Promipramine Hydrochloride, Lot Number: 050401, Specification: 25mg/tablet, produced by Shanghai Pharmaceutical Group Co., Ltd. According to the human clinical dose, it is converted into a dose of mouse H according to the body surface area, and is prepared by using pure water to prepare 0.75 mg/inl.

Naofukang (piracetam), batch number: 040302, specification: 0.4g/tablet, produced by Guangdong Kangbotong Pharmaceutical Industry Co., Ltd. According to the human clinical dose, the daily dose of the mouse is converted according to the body surface area, and 50 mg/ral is prepared by using pure water at the time of use.

The above positive control materials are all sealed and stored at room temperature.

4 experimental animals and their management

4. 1 experimental animals

Line: KM Mouse Grade: Grade 1 Weight: 24- 30g

Gender: Male Number of animals: 288

Supply unit: Experimental Animal Research Center, Chengdu University of Traditional Chinese Medicine

License number: SCXK (川) 2004-11

4.2 Feeding management of experimental animals: 4.2. 1 Animal room:

First Class Animal House, Sichuan Province Laboratory Animal Use License No.: 006,

Temperature · 18- 25 Ό Relative humidity: 50-70%

Ventilation: New wind Light conditions: 12/12 light and dark alternate

4.2.2 Drinking water: pure water, made by Sichuan Natural Medicine Research Institute.

Pure water preparation device: Reverse osmosis ion exchange system, Institute instrument number: 117.

4.2.3 Feed: Full-price mouse particles are fed, purchased from the Sichuan Provincial Experimental Animals Committee.

5, the main experimental equipment:

Standard mouse swimming pool (water tank height 30cm, length 30cm, 30cm)

Mouse Jumping Test Chamber, DTT-2, Institute of Materia Medica, Chinese Academy of Medical Sciences

Stopwatch, etc.

5, the main experimental reagents

Scopolamine, lot number: 4A18004, specification: 0.3mg/ml, Shanghai Hefeng Pharmaceutical Co., Ltd. Test Example 1 Screening test of the puzzle effect of the present invention - a mouse platform test

After the mouse plague was qualified, the computer (PEMS2.1 software) completely divided the mice into 12 groups, which were blank control group (saline group), positive (brain rehabilitation) control group, model control group and this group. Invention drug three The high, medium and low dose groups of the test articles, 12 mice in each group, were divided into conventional words. The drug was administered to the corresponding group (dosing volume of 0.2 ml/10 g) once a day. After 7 days, the platform training was carried out. 45 min before the training, each group of mice was intragastrically administered once, 30 min before the training, except for the blank control group, the other groups of mice ip scopolamine 2. 5 mg / kg, blank control group ip the same volume of normal saline . After the injection of lOniin, the test animals were placed in the reaction chamber for 3 minutes, and then immediately passed the 36V alternating current, training 5πΰη, and the number of times the animals were subjected to electric shock was recorded as the academic achievement. Animals were routinely taken after removal. After 24 hours, the test was repeated (24-hour memory retention test), the number of animals that were shocked, the latency of the first jump off the platform and the total number of errors within 3rain. The above test (long-term memory retention test) was repeated 6 days later, data was recorded and statistical analysis was performed.

1. 1 screening test for the role of puzzles - mouse platform test

The results are shown in Tables 2 and 3 and Figures 1 and 2.

Table 2 Puzzle drug screening - mouse platform test (24 hours memory retention) results, T + sd group dose C mg / kg) number of animals number of errors (times) number of shocks (times) mouse platform lurking

Ig- (only) period (S) blank control group saline 12 0. 17±0. 39** 0. 17 persons 162. 3 ±47. 9**

0. 39**

Model control group saline 12 1. 33±0. 65 1. 42±0. 79 47. 5±61. 0 positive (brain rehabilitation) control group 50 11 0. 64±1. 29* 1. 45±1 81 147. 8 soil 61. 3* The drug A-low dose group of the invention 30 10 0. 50 ±0. 97** 0. 8±0. 79 128. 8±82. 5 The drug A-medium dosage of the invention Group 60 12 0. 33 ±0. 49** 0. 83 ±0. 72 126. 8±81. 0 Drug A-high dose group 120 12 0. 75 ±0. 97 1. 50± 1. 51 101. 9±84. 1 The drug of the invention B-low dose group 30 12 0. 25 ±0. 45** 0. 5±0. 67* 148. 6 ±72. 5* The drug B-medium dosage group of the invention 60 12 0. 00 ±0. 00** 0. 58±0. 9 180. 0±0. 0** The drug of the present invention B-high dose group 120 11 0. 45 ±0. 93** 0. 64± 0. 67 137. 7±73. 4 The drug of the present invention C-low dose group 30 12 0. 33 ±0. 65** 1. 08±1. 31 140. 6 ±72. 3 C-medium dosage of the drug of the present invention Group 60 12 0. 58 ±0. 67* 0. 67 ±0. 65 116. 6±75. 9 The drug of the present invention C-high dose group 120 12 0. 50 ±0. 67** 1. 17 ±1. 47 111. 2±74. 0 Note: Analysis was performed using SPSS11. 0 One-way A OVA-LSD-TUKEY.

"*" indicates comparison with the model control group, p<0.05. "**" indicates comparison with the model control group, p<0.01. Table 3: Puzzle drug screening - mouse platform test (long-term memory retention) Results , Γd sd

(mg/kg) Number of animals Mouse platform latency period Group Mistakes (times) Sniper times (times)

Ig. (only) (s) blank control group saline 12 0. 42 ±1· 44 0. 58 ±1. 44 165. 1±51. 4** model control saline 11 0. 45 ±0. 69 1. 10 ±0. 94 115. 7±75. 4 Positive (brain rehabilitation) control group 50 11 0. 00 ±0. 00 0. 09±0. 31** 180. 0+0. 00** Inventive drug A-low dose group 30 9 0. 00 ±0. 00 0. 11 ±0. 33** 163. 1±50. 7* The drug A-medium dose group of the invention 60 12 0. 17±0· 58 0. 25 ±0. 62** 165. 8 ±49. 1** Difficult A-high dose group 120 12 0. 00 ±0. 00 0. 09±0. 30** 180. 0±0. 0** The drug of the present invention B-low dose group 30 11 0. 08 ±0. 29 0. 08 ±0. 29** 166. 0±48. 5** The drug of the invention B-medium dose group 60 12 0. 00±0. 00 0. 00+0. 00** 180. 0±0. 0** The drug of the present invention B-high dose group 120 11 0. 00 ±0. 00 0. 09+0. 30** 180 0±0. 0** The drug of the present invention C-low dose group 30 12 0. 00±0· 00 0. 08 ±0. 29** 180. 0±0. 0** The drug of the invention C-medium dose Group 60 11 0. 09 ±0. 30 0. 18 ±0. 60** 168. 1±39. 5** The drug of the invention C-high dose group 120 12 0. 58 ±0. 67 0. 83 ±0 94 122. 7±71. 2 Note: HI'SPSSll. O One-way ANOVA- LSD analysis.

"*" indicates that compared with the model control group, p<0.05; "**" indicates that compared with the cast type control group, p<0.01 test the mouse platform test to evaluate the platform latency, the number of errors and the number of shocks. Among them, the incubation period is the main indicator, which can objectively and accurately judge the memory ability of mice. The other two are secondary indicators. The judgment is relatively subjective and the evaluation efficiency is low, so it is used as a secondary indicator.

The experimental results showed that: in the 24-hour memory retention test results of the mouse platform test, SPSS1. 0 One-way ANOVA-LSD-TUKEY was used for data analysis, and the blank control group compared with the model control group, the number of mouse errors, the number of electric shocks and There were significant differences in the indexes of the platform latency (P<0.01). At the same time, there were significant differences between the positive (brain rehabilitation) control group and the model control group in terms of the number of mouse errors and the platform latency (ρ< 0. 05), indicating that the experimental model is established and the experimental evaluation system is reliable (see Table 2). Among the three tested samples of the present invention, 50.37% of the low dose (30 mg/kg) group was significantly different from the model control group (p<0.05); 50. 37% of the medium dose (6013⁄4/13⁄4) group Compared with the model control group, there was a significant difference (p<0.01). It can be seen that, under the experimental conditions, among the three tested samples, the test sample B-50. 37% in the 24-hour memory retention test, keeping the mouse memory, improving the intelligence compared with the other two samples Has a more prominent effect.

In the long-term memory retention test, SPSS11.0 One-way ANOVA-LSD was used for data analysis. The experimental system was still established with reference to the mouse platform latency (see Table 3). In the dose group of the drug of the present invention, except for the C high dose group (120 mg/k _g ), there was a significant difference (ρ<0.01) compared with the model control group, indicating that each sample was administered after 1 day. It has a positive effect on the long-term memory retention of mouse jumping.

In summary, 50.37% of the mice have a memory retention effect in the short- and medium-term of the mice, which is more effective than the other samples.

Test Example 2 Antidepressant Screening Test of the Drug of the Invention - Forced Mouse Swimming Test (Acquired Despair Model) After the mouse was quarantined, the computer (PEMS 2.1 software) completely randomized the mice into 12 groups, respectively Control group (saline group), positive (brain rehabilitation) control group, model control group and high, medium and low dose groups of three test articles of the present invention, 12 mice in each group, caged conventional words . The corresponding group of drugs (administered volume 0.2ral/10g) was administered once a day. Two days before the test, the model mice were given lOmin swimming training once a day. The mice were fasted for 16 h before the test. After 30 minutes of re-administration for the first 30 minutes of the evaluation, a single mouse was placed in the swimming pool water for 6 minutes, and the cumulative time of the mice within 4 minutes after the recording was recorded (test on the 9th day). (Do not move means that the mouse stops struggling in the water, or the animal is floating, only the small limbs move to keep the head floating on the water surface), and the above test is repeated once every 7 days (the 15th day test). Data was recorded and statistically analyzed.

The experimental results were analyzed by ANOVA and X ² test using SPSS statistical software.

The results are shown in Tables 4 and 5, Figure 3, and Figure 4. Antidepressant screening-forced swimming test (day 9) Results, sd group dose (mg/kg) ig. Number of animals (only) Mice immobility time (s> blank control group equal volume of saline 10 1.5±2.8** model control group equal volume of normal saline 10 32.4 ± 30.3 positive ( imipramine) control group 0.75 10 6.8 ± 3.9 * The drug of the invention A - low dose group 30 10 25.7 ± 25.1 The present invention A- Dosage group 60 10 15.1±22.5 The drug A-high dose group of the invention 120 10 7.0±12.9* The drug B-low dose group of the invention 30 10 28.1±29.0 The drug B-middle dose group of the invention 60 10 34· 4 ±18.6 Inventive drug B-high dose group 120 10 3.2±5.6* The drug of the invention C-low dose group 30 10 67.3 ±38.9# The drug of the invention C-medium dose group 60 10 33.1±29.2 The drug of the invention C-髙 dose group 120 9 18.2±25.4

Note: In order to reduce the dispersion of data, each group of data is removed from the highest and lowest values and statistically processed. Therefore, the number of samples in each group is reduced by two compared with the actual sample. Analysis was performed using SPSS 11.0 One-way ANOVA-LSD.

"*" indicates p<0.05 compared with the model control group; "**" indicates comparison with the model control group, p<0.01; "#" indicates that compared with the model control group, p<0.05, but the mean value is greater than the model control. group.

Table 5 Antidepressant Screening - Forced Mouse Swimming Test (Day 15) Results, IT soil sd group dose (mg/kg) ig. Number of animals (only) Mouse immobility time (S) Blank control group, etc. Volumetric saline 10 61.3±42.8** Model control group equal volume of normal saline 10 122.5 ± 40.6 imipramine (positive) control group 0.75 10 98.1 ± 48.7 Drug A-low dose group of the invention 30. 10 82.2 ± 35.3 * The present invention Drug A-medium dose group 60 10 117.6±34.3 Drug A-high dose group of the invention 120 10 36.3 ±32.0** Drug B-low dose group of the invention 30 10 88.9 ±28.9 Drug B-middle dose group of the invention 60 10 128.8 ±46.3 Drug B-high dose group of the invention 120 10 81.4±48.4* The drug of the invention C-low dose group 30 10 115.6±45.4 The drug of the invention C-middle dose group 60 10 89.1±32.9 The C-high dose group of the invention 120 8 80.8 ±39.9* Note: In order to reduce the discreteness of the data, each group removes the highest and lowest value samples and performs statistical processing. Adopt SPSS11.0

One-way ANOVA- LSD analysis.

"*" indicates p<0.05 compared with the model control group; "**" indicates p<0.01 compared with the model control group.

The experimental results showed that: SPSS11.0 One-way ANOVA-LSD was used for data analysis on the 9th day of the forced swimming test in mice. The blank control group and the positive (imiprozine hydrochloride) control group were compared with the model control group. There were extremely significant and significant differences (p<0.01 and p<0.05), indicating that the experimental model was established and the experimental evaluation system was reliable (see Table 4). The 21.11%, 50.37% high dose (120mg/kg) group was significantly different from the model control group (p<0.05). There was no difference between the other sample dose groups and the model control group. It can be seen that under the experimental conditions, 21.11% and 50.37% of the three test samples showed a confrontation in the test on the 9th day under the condition of continuous intragastric administration of 120 mg/kg per day. The role of the mouse acquired despair model state.

Data analysis was performed using SPSS11.0 One-way ANOVA-LSD on the 15th day of the test: There was a significant difference between the white control group and the model control group ( _P < 0.01), while the positive control substance (imipramine hydrochloride) had a tendency to alleviate the state of desperate depression in mice, but there was no data between the experimental groups. Statistically significant (see Table 5), this may be related to the pharmacodynamic profile of imipramine itself. In the drug dosage group of the present invention, the 21.11% low dose group, the 50.37% sputum dose group, and the 72.50% sputum dose group were significantly different from the model control group (p<0.05); 21. The 11% sputum dose group was significantly different from the model control group (p<0.01). This indicates that each test sample has a role in combating depression in mice under certain doses and a certain administration time, and 21.11% of the high doses seem to have a longer time. Stable against the pharmacodynamic effects of desperate depressive symptoms in mice, therefore, in the three samples of the present invention, the screening of antidepressant function was superior to 21.11%.

In summary, it can be seen that the three samples of the present invention have certain functions to promote the mental and anti-depressive functions of the model mice, but the efficacy is not the same.

In terms of puzzle: For the 24-hour memory retention function, 50. 37% is effective at 30 mg/kg, and within 30 mg/kg to 60rag/kg, there is a dose-effect relationship, and the onset of effect seems to have decreased. Space; long-term memory retention, the sample three-dose group showed good pharmacodynamic effects, in which the incubation period of the middle and high dose groups reached more than 3min, no obvious mistakes occurred, and the positive control drug brain complex Compared with Kang, the dose is lower and the effect is more clear. The results of the two-hour evaluation of the 24-hour and long-term memory retention ability were basically the same, indicating the effectiveness of the drug of the present invention in promoting the memory ability of the mouse, and indirectly indicating the reproducibility of the experimental study.

In terms of antidepressant: Under the experimental conditions, the experimental system was established. The study found that in the test evaluation on the 9th day, 21.11% and 50.37% of the high-dose group were effective against the model of despair depression in mice caused by forced swimming, and its pharmacodynamic effect was comparable to that of imipramine hydrochloride. . In the 15th day of the evaluation, 21. 11% still maintained a good antidepressant effect, and the low dose group also showed a certain antidepressant effect. The high dose group had a very significant difference compared with the model group. A certain time-effect relationship, the efficacy seems to be better than the imipramine control group at this dose. 50. The 37% high dose group remained as strong as the test on day 9. The results of the two evaluations were basically the same, which also confirmed the reliability of the study.

In summary, the effect of the puzzle: the best efficacy sample: 50. 37%

Antidepressant effects: Best Pharmacodynamics: 21. 11%

According to the seventh paragraph of the Chinese Pharmacopoeia 2005 edition of the second appendix XIX A, the determination of the content of the drug substance and preparation, the range should be 80% - 120% of the test concentration; the uniformity of the preparation content, the scope should be the test concentration 70%-130%. The optimal range of pharmacodynamic effects of the puzzle in this experiment is: 50% ± 0.1, the best anti-depression sample range: 20% ± 0.04.

main reference

[1] Xie Xiuqiong, editor, Development and application of new Chinese medicine preparations, People's Medical Publishing House, 1994

[2] Xu Shuyun and other editors, Pharmacological Experimental Methodology, Third Edition, People's Medical Publishing House, 2001

[3] Li Yikui, Editor-in-Chief, Chinese Medicine Pharmacology Experimental Methodology, Shanghai Science and Technology Press, 1991

Claims

Claim

A extract of P. oleracea, characterized in that the total percentage of total saponins of P. oleracea is in the range of Bacoside A, which is 10% to 80% of the extract.

The extract of P. oleracea according to claim 1, wherein the total percentage of total saponins of P. oleracea is in terms of Bacoside A, which is 10 ° / of the extract. ~40%.

The extract of P. oleracea according to claim 2, wherein the total percentage of total saponins of P. oleracea is from 10% to 30% by weight of Bacoside A.

The extract of P. oleracea according to claim 3, wherein the total amount of the total saponin of the scutellaria is Bacoside A, which is 20% ± 0.04 of the extract.

The extract of P. oleracea according to claim 1, wherein the total content of total saponins of P. oleracea is from 40% to 70% by weight of Bacoside A.

The extract of the scutellaria sinensis according to claim 5, wherein the total saponin content of the saponin is determined by Bacoside A, which is 50% ± 0.1 of the extract.

The extract of Pseudomonas sinensis according to any one of claims 1 to 6, wherein the false purslane is derived from the whole grass of the genus Scrophulariae Bacopa monnieri L.

The extract of the scutellaria chinensis according to any one of claims 1 to 7, wherein it is extracted by the following method: taking the whole grass of the dried squid Bacopa monnieri L., 40%~ 80% ethanol extraction, the extract is directly concentrated and dried to obtain the extract of P. oleracea; or after the extract is concentrated, the macroporous adsorption resin is adsorbed, and then eluted with 10% to 95% ethanol, and the eluate is collected and dried. That is.

A method of producing the extract of the scutellaria chinensis according to any one of claims 1 to 8, which comprises the steps of:

a, weigh the whole grass of Bacopa monnieri L., add 40% ~ 80% ethanol extraction, the extract is concentrated to obtain a concentrated solution;

10. A method of preparing a extract of P. oleracea according to claim 9, wherein: the ethanol elution as described in step b is: eluted with 60 to 95% ethanol.

11. The method according to claim 9, wherein the elution of the ethanol in step b is: first eluting with 10% to 60% ethanol, and then using 60%~ Eluted with 95% ethanol.

12. Use of the extract of Pseudomonas sinensis according to any one of claims 1 to 4 for the preparation of a medicament for antidepressant.

13. Use of the extract of P. oleracea according to claims 1, 5, 6 for the preparation of a medicament having a puzzle.

A pharmaceutical composition which comprises an effective amount of the extract of the scutellaria purslane according to any one of claims 1 to 8, which is prepared by adding a pharmaceutically acceptable excipient or an auxiliary component. .

The pharmaceutical composition according to claim 14, wherein the preparation is an oral preparation or an injection.

16. Use of a pharmaceutical composition according to claim 14 or 15 in the manufacture of a medicament for antidepressant.

17. The use according to claim 16, wherein: the extract of the scutellaria sinensis extract containing the total glucosides of the scutellaria sinensis is based on Bacoside A, which accounts for 10% of the extract. ~80%.

18. Use according to claim 17, characterized in that the pseudo-dentate width extract of the medicament contains the total percentage of total saponins of the purslane saponins in the range of 10 to 40% by weight of Bacoside A.

The use according to claim 18, characterized in that: in the medicament, the weight percentage of the total saponin of the scutellaria sinensis extract is 20% ± 0.04 in terms of Bacoside A.

20. Use of a pharmaceutical composition according to claim 14 or 15 for the preparation of a nootropic drug.

The use according to claim 20, characterized in that: the drug contains a total percentage of total saponins of P. oleracea extract in terms of Bacoside A, which accounts for 10% of the extract. 80%.

The use according to claim 21, characterized in that: in the medicament, the weight percentage of the total extract of the false purslane containing the purslane is 40 to 70% by weight of Bacoside A.

The use of the drug according to claim 22, wherein: the weight percentage of the total saponin of the scutellaria sinensis extract containing the saponin of the purslane is 50% ± 0.1% by weight of Bacoside A. .