CN104258193B - A kind of pharmaceutical composition for treating neurasthenia and prevention senile dementia - Google Patents

Abstract

The invention discloses a pharmaceutical composition for treating neurasthenia and preventing senile dementia. 10%, and the balance is auxiliary materials. The medicine provided by the invention is helpful for the prolongation of the sleep time of the elderly, coix seed lactone can prevent cancer and inhibit the proliferation of various cancer cells, and the combination of the three can effectively prevent and control the occurrence and cure of senile dementia, and give the aging more More and more serious world brings a safe and effective botanical composition of pure plant extracts. The prescriptions adopted in the present invention have reasonable compatibility, and the raw materials are all from natural plants. At the same time, the preparation method of the traditional Chinese medicine composition provided by the present invention has strong operability, especially according to the extraction process related to the chemical properties of the active ingredients of various traditional Chinese medicines, the obtained extracts are effective. The composition content is high, and it has the advantages of high drug efficacy, less clinical dosage, etc., and the process is stable, which is suitable for large-scale industrial production.

Description

A pharmaceutical composition for treating neurasthenia and preventing senile dementia

technical field

The invention belongs to the technical field of biomedicine and relates to a pharmaceutical composition for treating neurasthenia and preventing senile dementia.

Background technique

With the development of my country's economy and the implementation of population policy, the age structure of my country's population has undergone great changes. China's life expectancy rose from 68.4 years in 1990 to 73.8 years in 2008, and the proportion of the elderly population in the total population is increasing. According to the data provided by the Organization for Economic Cooperation and Development, relative to China's total population, it is estimated that The proportion of the population over the age of 65 will rise from 7.8% in 2005 to 16.3% in 2030.

The researchers found that after the shark's brain tissue was severely injured, it can automatically recover its function, because the shark's brain tissue contains a substance called "nervous acid", which can repair the nerve information transmission channel of the brain - nerve fibers, and promote nerve cells to recover. regeneration. Today, nervonic acid (NA) has been recognized by scientists from all over the world as the core natural active ingredient of brain nerve fibers and nerve cells. Neural acid can also effectively treat brain diseases such as senile dementia, cerebral palsy, brain atrophy, memory loss, insomnia and forgetfulness. Humans are different from sharks. Humans cannot synthesize nervonic acid by themselves. As they age or the brain is traumatized, it may cause the attenuation of nervonic acid and lead to disease.

Extracting nervonic acid from sharks must kill sharks and destroy the ecological balance of the ocean. With the increasing calls of international organizations to ban the killing of sharks, countries such as Europe and the United States attach great importance to excavating nervonic acid resources from plants. Research has found that it contains 2 There are more than ten kinds of plants with more than 100% nervonic acid, among them, the unique single plant in China - Garlic fruit has the highest nervonic acid content, and it is as high as 62.6%. Its development value and economic value are huge, and it can be called "the golden fruit of plants". ".

Malania oleifera Chun et Lee is a plant of the genus Malania in the Olacaceae family . Also known as Shantong fruit, after Malan. It belongs to the second-class national protected plants. It is distributed in the southeastern part of Yunnan such as Funing, Guangnan and Wenshan. The seed kernel of garlic fruit is rich in oil and is an ideal raw material for the synthesis of muskone (muscine) . The oil content of garlic fruit seeds is as high as 50-60%, and the fatty acid after saponification contains 40-50% tetracos-15-enoic acid, that is, nervonic acid, which helps to restore the activity of nerve endings, promote nerve cell growth and Development, has anti-senile dementia effect.

According to statistics, there are now 24 million dementia patients in the world. In my country, the number of patients with Alzheimer's disease has reached more than 5 million, and there are 300,000 new cases every year. The pathogenesis of Alzheimer's disease and the research and development of new drugs are being paid great attention by countries all over the world. Therefore, using garlic fruit as a raw material to develop edible oils, health products and medicines for the treatment of senile dementia has great market potential and medical value.

Coix seed is the mature kernel of Coix lacryma-jobi L., a perennial herb of the Poaceae family. Coix seed is produced in most parts of China. Because of its high nutritional value, Coix seed is known as the world's The king of grasses. Coix seed contains about 2.7% of coix seed oil, and Coix seed contains 16.2% protein, 4.65% fat, 79.17% carbohydrate, and a small amount of vitamin B ₁ . Coix seed contains a variety of amino acids (including leucine, lysine, arginine, tyrosine, etc.) and also contains coixin, coixin, triterpenoids and other substances. The effective ingredients in coix seed that inhibit the proliferation of various tumor cells are coixin and coixenolactone. These compounds also have the functions of enhancing hormone regulation and promoting the immune system and enzyme system, and can also promote cellular immunity and humoral immunity. A large number of animal experiments and clinics have confirmed that coix seed oil has the effect of enhancing immunity and anti-tumor. Job's tears are planted all over my country. Yunnan Province, represented by Shizong County, Qujing City, has rich resources of Job's tears and is one of the important growing areas of Job's tears in my country. Coix seed is rich in nutrition and has good medicinal value such as anti-tumor, promoting blood circulation and removing blood stasis. Some coix seed products have appeared in the market one after another, such as coix seed health wine, coix seed vinegar, coix seed powder, coix seed tea, etc., which are very popular. The majority of consumers welcome. However, compared with foreign barley products, the development of domestic barley products is relatively insufficient, and the utilization of barley is basically still in people's diet, and there is a serious shortage of deep-processed products.

Tofu fruit, also known as sow fruit, tofu fruit extract is tofu fruit smoothie (helicid) , p-formaldehyde benzene-O-β-D-allopyranoside, also known as Kunming Shenshuai fruit, is a wild plant in Yunnan The fruit of ophthalmic radish tree (Helieiaerra6c:Hook.f) (commonly known as bean curd fruit), its chemical structure is similar to gastrodin. Pharmacological and clinical studies have shown that the effect of tofu smoothie on the central nervous system is similar to that of gastrodin, but its sedative and analgesic effects are stronger than gastrodin, and its therapeutic effect on headache, dizziness, and sleep disorders caused by neurosis is rapid. At the same time, it has a good improvement and regulation effect on memory and thinking power, and has broad application prospects. Sow fruit is mainly distributed in the south and southwestern parts of Yunnan Province, with the largest distribution in Pu'er City and Honghe Prefecture. Most of them are in a wild state, and they are in urgent need of artificial breeding protection and development and utilization of deep-processed products.

The present invention takes three kinds of medicinal plants concentratedly distributed or cultivated in Yunnan Province as the research objects, and uses phytochemical extraction methods to extract the active ingredients of preventing Alzheimer's disease, anti-depression and anti-tumor from these three plants, and uses medicament It is prepared into traditional Chinese medicine health-care tablets or capsules by scientific means, and provides a safe and effective traditional Chinese medicine health-care product for middle-aged and elderly patients.

Contents of the invention

In order to solve the problems in the prior art that raw materials for preventing senile dementia, antidepressant and antitumor drugs are difficult to obtain, and current drugs have side effects, the present invention provides a pharmaceutical composition for treating neurasthenia and preventing senile dementia, using pure Plant raw materials are used to prepare anti-senile dementia, neurasthenia and anti-tumor drugs.

The present invention is achieved through the following technical solutions: a pharmaceutical composition for treating neurasthenia and preventing senile dementia, characterized in that it consists of the following components in mass percentage:

Garlic Neural Acid 1～10%,

Tofu fruit glycosides 1~10%,

Coix seed lactone 1～10%,

The balance is excipients.

The auxiliary materials include conventional dextrin, lactose, sodium carboxymethyl starch, microcrystalline cellulose, starch and hydroxypropyl cellulose in the field of pharmacy.

Described garlic nervine acid extracts, purifies according to the following steps:

(1) Remove the shell and epidermis of the garlic seeds, and pound them with a phosphate buffer solution with a concentration of 20mM and a pH of 6.0 to 7.0 at 3 to 4°C according to the solid-to-liquid ratio g/mL of 1 to 10:5 to 50. After being crushed, it was subjected to ultrasonic crushing treatment for 30-50 minutes, extracted at 3-4°C overnight, then filtered, and then centrifuged at 5000-8000 r/min at 3-4°C for 30-40 minutes. Repeat twice, remove the supernatant to get the residue;

(2) Add petroleum ether to the residue obtained in step (1) according to the solid-to-liquid ratio g/mL=1～5:5～20 for stirring, then let it stand for stratification, collect the filtrate after filtration, and soak the filter residue repeatedly with the filtrate 2-3 times, filter after soaking for 30-60 minutes each time, combine the filtrates, and then perform rotary evaporation at 40-50°C to remove petroleum ether, and obtain garlic fruit oil;

(3) Weigh the silica gel G according to the mass ratio of the sample amount: the amount of silica gel = 1 ~ 10: 10 ~ 100, first add petroleum ether to dissolve the silica gel G according to the solid-liquid ratio g/mL = 1 ~ 5: 2 ~ 10, and then After stirring for 20-30 minutes, pour it into the chromatographic column. After the silica gel chromatographic column does not contain air bubbles and the sedimentation is stable, the garlic fruit oil obtained in step (2) is mixed according to the solid-liquid ratio g/mL=1-5:1 ~10 After adding petroleum ether to dissolve, put it into the upper layer of the silica gel chromatography column, and use petroleum ether-ethyl acetate elution system to carry out gradient elution, according to petroleum ether dissolved with garlic fruit oil:ethyl acetate=50:1～30 : 1 (V/V) fraction, the fraction was concentrated under reduced pressure at 40-50°C to obtain colorless blocky crystals, which is allicin.

The nervonic acid standard and the colorless crystal were spotted on the same thin-layer plate by thin-layer chromatography, developed, and compared. As a result, the colorless crystal and the nervonic acid standard showed the same color at the same position on the same thin-layer plate. Purple spots, and the same color spots and the same Rf value in three different unfolding systems, prove that the colorless crystal is nervonic acid. The content of nervous acid obtained by this method is greater than 95%.

The phosphate buffer in the step (1) is prepared according to the following operations: mix the disodium hydrogen phosphate solution with a concentration of 0.2M and the sodium dihydrogen phosphate solution with a concentration of 0.2M in a volume ratio of 12.3～61:87.7～39, That is, a phosphate buffer solution with a pH of 6.0 to 7.0 is obtained.

The supernatant of the step (1) is subjected to subsequent purification and separation to obtain protein precipitates.

The particle size of the silica gel G in the step (3) is 200-300 mesh.

The tofu fruit glycosides are extracted and purified according to the following method, through the following steps:

(1) Crush the bean curd seeds, pass through a 100-mesh sieve, then add ethanol with a volume concentration of 50-70%, cold-soak at 20-30°C for 30-60 minutes, then ultrasonically extract for 30-60 minutes, filter, and the filtrate is at 45-55 Carry out rotary evaporation and concentration at ℃, recover ethanol, and obtain sow fruit ethanol extract;

(2) Add pure water to the tofu fruit ethanol extract obtained in step (1) according to the solid-to-liquid ratio g/mL of 1-5:5-25 to dissolve, apply macroporous resin D101 type, and first elute with pure water to remove Water-soluble impurities such as polysaccharides and proteins, and then elute 4 to 5 retention volumes with ethanol with a volume concentration of 30 to 50%, collect the ethanol effluent with a volume concentration of 30 to 50% of the macroporous resin, concentrate under reduced pressure, and then Add 90-95% ethanol with a concentration of 2-3 times the volume of the concentrate, let stand at 3-4°C for 12-24 hours to precipitate crystals, and then filter out the crystals to obtain pure tofu fruit glycosides. The purity of the obtained tofu fruit glycosides is greater than 95%.

The coix seed lactone is extracted and purified according to the following method, through the following steps:

Crush coix seed, pass through a 100-mesh sieve, and then extract coix seed powder with supercritical carbon dioxide for 1-2 hours at a temperature of 35-40°C, a pressure of 37-40 MPa, and a _CO flow rate of 10-20 kg/h. Dissolve the extract in ethanol with a concentration of 70-95%, and then remove the ethanol by rotary evaporation to obtain yellow oil, which is coix seed lactone.

The above-mentioned garlic fruit nervonic acid, tofu glucoside, and coix seed lactone extracts are fully mixed in proportion, and then added with pharmaceutically acceptable auxiliary materials to prepare any commonly used oral preparations, such as capsules, granules, oral liquids, and pills or tablet.

The advantages and effects of the present invention: the medicine provided by the present invention is mainly a combination of small molecular compounds such as fatty acids and lactones. Through phytochemical extraction methods, active ingredients are extracted from several plants that are unique to Yunnan and grown in large areas. Among them Nervous acid has a significant anti-senile dementia effect. Tofu fruit glycosides can relieve the anxiety and tension of senile dementia patients and help the elderly to prolong their sleep time. Coix seed lactone can prevent cancer and inhibit the proliferation of various cancer cells. The combination of the three can effectively prevent and control the occurrence and cure of senile dementia, and bring a safe and effective plant composition of pure plant extracts to the world where the aging is getting worse. The prescriptions adopted in the present invention have reasonable compatibility, and the raw materials are all from natural plants. At the same time, the preparation method of the traditional Chinese medicine composition provided by the present invention has strong operability, especially according to the extraction process related to the chemical properties of the active ingredients of various traditional Chinese medicines, the obtained extracts are effective. The composition content is high, and it has the advantages of high drug efficacy, less clinical dosage, etc., and the process is stable, which is suitable for large-scale industrial production.

detailed description

The present invention is further illustrated below by way of examples.

Example 1

The pharmaceutical composition for treating neurasthenia and preventing senile dementia consists of the following components in mass percentage:

Garlic Neural Acid 2%,

Tofu fruit glycosides 6%,

Coixenolide 5%,

Excipients 87%,

The auxiliary materials are dextrin, lactose, sodium carboxymethyl starch, microcrystalline cellulose, and starch.

Wherein the garlic fruit nerve acid is extracted and purified according to the following steps:

(1) Remove the shell and epidermis of the garlic fruit seeds, according to the solid-liquid ratio g/mL of 10:50, mash with phosphate buffer solution with a concentration of 20mM and pH=6.0 at 4°C, and then ultrasonically pulverize 40min, extract overnight at 3°C, then filter with 4 layers of gauze, then centrifuge at 6000 r/min for 40min at 3°C, repeat twice, remove the supernatant to obtain the residue; The salt buffer solution is prepared according to the following operations: mix the disodium hydrogen phosphate solution with a concentration of 0.2M and the sodium dihydrogen phosphate solution with a concentration of 0.2M at a volume ratio of 61:39 to obtain a phosphate buffer solution with pH=7.0; The supernatant is purified and separated to obtain protein precipitation;

(2) Add petroleum ether to the residue obtained in step (1) according to the solid-to-liquid ratio g/mL=3:20 for stirring, then let it stand for stratification, collect the filtrate after filtration, and then soak the filter residue 3 times with the filtrate repeatedly, each Filter after soaking for 50 minutes for the first time, combine the filtrates, and then carry out rotary evaporation at 40°C to remove petroleum ether to obtain light yellow garlic fruit oil;

(3) Weigh the silica gel G with a particle size of 300 mesh according to the mass ratio of sample amount: silica gel amount = 5:60, first add petroleum ether to dissolve the silica gel G according to the solid-liquid ratio g/mL=3:8, and then stir for 25 minutes. Pour it into the chromatographic column. After the silica gel chromatographic column does not contain air bubbles and the sedimentation is stable, the garlic fruit oil obtained in step (2) is dissolved in petroleum ether according to the solid-to-liquid ratio g/mL=5:10, and then put into The upper layer of the silica gel chromatography column is gradiently eluted with petroleum ether-ethyl acetate elution system, according to the fraction of petroleum ether dissolved with garlic fruit oil:ethyl acetate=40:1 (V/V), at below 50°C Concentrate the fractions under reduced pressure to obtain colorless blocky crystals, which are allicin.

The nervonic acid standard and the colorless crystal were spotted on the same thin-layer plate by thin-layer chromatography, developed, and compared. As a result, the colorless crystal and the nervonic acid standard showed the same color at the same position on the same thin-layer plate. Purple spots, and the same color spots and the same Rf value in three different unfolding systems, prove that the colorless crystal is nervonic acid. The content of nervous acid obtained by this method is greater than 95%. The yield of nervonic acid is 10.462% of that of garlic fruit oil.

Wherein the tofu glucoside is extracted and purified according to the following method, and undergoes the following steps:

(1) Crush the bean curd seeds, pass through a 100-mesh sieve, then add ethanol with a volume concentration of 70% and soak for 40 minutes at 25°C, then ultrasonically extract for 60 minutes, filter, and the filtrate is concentrated by rotary evaporation at 45°C to recover ethanol. Obtain the ethanol extract of sow fruit;

(2) Add pure water to the ethanol extract of bean curd fruit obtained in step (1) according to the solid-to-liquid ratio g/mL of 5:25 to dissolve, apply macroporous resin D101 type, and first elute with pure water to remove polysaccharides, proteins, etc. Water-soluble impurities, then elute 5 retention volumes with ethanol with a volume concentration of 50%, collect the ethanol effluent with a volume concentration of 30% of the macroporous resin, concentrate under reduced pressure, and then add the concentration of 3 times the volume of the concentrate to be 92 % ethanol, stand at 3°C for 20h to precipitate crystals, and then filter out the crystals to obtain pure tofu fruit glycosides. The purity of the obtained tofu glucoside is greater than 95%, and the yield of the tofu glucoside is 1.06% of that of sow fruit seeds.

Among them, coix seed lactone is extracted and purified according to the following method, and undergoes the following steps:

Crush coix seed, pass it through a 100-mesh sieve, and then conduct supercritical carbon dioxide extraction for 1 h under the conditions of temperature 40°C, pressure 37 MPa, and CO ₂ flow rate 15 kg/h, and use ethanol with a volume concentration of 70% to dissolve and extract substance, and then the ethanol was removed by rotary evaporation to obtain yellow oil, which was coix seed lactone. The extraction yield of coix seed lactone was 5.3%.

The above-mentioned allicin, tofu glucoside and coix seed lactone extracts are fully mixed in proportion, and then pharmaceutically acceptable auxiliary materials are added to prepare any commonly used capsules.

Example 2

The pharmaceutical composition for treating neurasthenia and preventing senile dementia consists of the following components in mass percentage:

Garlic Nervous Acid 1%,

Tofu glucoside 10%,

Coix Seed Lactone 10%,

Excipients 79%,

The auxiliary materials are dextrin, lactose, sodium carboxymethyl starch, microcrystalline cellulose, starch, and hydroxypropyl cellulose.

Wherein the garlic fruit nerve acid is extracted and purified according to the following steps:

(1) Remove the shell and epidermis of the garlic fruit seeds, according to the solid-to-liquid ratio g/mL of 1:30, mash with phosphate buffer solution with a concentration of 20mM and pH=7.0 at 3°C, and then ultrasonically pulverize 50min, extract overnight at 4°C, then filter with 4 layers of gauze, then centrifuge at 5000r/min at 4°C for 35min, repeat twice, remove the supernatant to obtain the residue; the phosphate buffer The solution is prepared according to the following operations: mix the disodium hydrogen phosphate solution with a concentration of 0.2M and the sodium dihydrogen phosphate solution with a concentration of 0.2M at a volume ratio of 12.3:87.7 to obtain a phosphate buffer solution with pH=6.0; the supernatant After subsequent purification and separation, protein precipitation is obtained;

(2) Add petroleum ether to the residue obtained in step (1) according to the solid-to-liquid ratio g/mL=5:10 for stirring, then let it stand for stratification, collect the filtrate after filtration, and then soak the filter residue twice with the filtrate, each time After soaking for the first time for 30 minutes, filter, combine the filtrates, and then carry out rotary evaporation at 45°C to remove petroleum ether to obtain light yellow garlic fruit oil;

(3) Weigh the silica gel G with a particle size of 250 mesh according to the mass ratio of the sample amount: the amount of silica gel = 10:100, first add petroleum ether to dissolve the silica gel G according to the solid-liquid ratio g/mL=5:2, and then stir for 20 minutes. Pour it into the chromatographic column. After the silica gel chromatographic column does not contain air bubbles and the sedimentation is stable, the garlic fruit oil obtained in step (2) is dissolved in petroleum ether according to the solid-to-liquid ratio g/mL=3:1, and then put into The upper layer of the silica gel chromatography column is gradiently eluted with petroleum ether-ethyl acetate elution system, according to the fraction of petroleum ether dissolved with garlic fruit oil: ethyl acetate = 50:1 (V/V), at below 45°C Concentrate the fractions under reduced pressure to obtain colorless blocky crystals, which are allicin.

The nervonic acid standard and the colorless crystal were spotted on the same thin-layer plate by thin-layer chromatography, developed, and compared. As a result, the colorless crystal and the nervonic acid standard showed the same color at the same position on the same thin-layer plate. Purple spots, and the same color spots and the same Rf value in three different unfolding systems, prove that the colorless crystal is nervonic acid. The content of nervous acid obtained by this method is greater than 95%. The yield of nervonic acid is 11.054% of that of garlic fruit oil.

Wherein the tofu glucoside is extracted and purified according to the following method, and undergoes the following steps:

(1) Crush the bean curd seeds, pass through a 100-mesh sieve, then add ethanol with a volume concentration of 60%, soak at 20°C for 60 minutes, then extract with ultrasonic waves for 45 minutes, filter, and the filtrate is concentrated by rotary evaporation at 50°C to recover ethanol. Obtain the ethanol extract of sow fruit;

(2) Add pure water to the tofu fruit ethanol extract obtained in step (1) according to the solid-to-liquid ratio g/mL of 3:19 to dissolve, apply macroporous resin D101 type, and first elute with pure water to remove polysaccharides, proteins, etc. Water-soluble impurities, then elute 4 retention volumes with ethanol with a volume concentration of 40%, collect the ethanol effluent with a volume concentration of 40% of the macroporous resin, concentrate under reduced pressure, and then add the concentration of 2 times the volume of the concentrate to 95 % ethanol, stand at 4°C for 24 hours to precipitate crystals, and then filter out the crystals to obtain pure tofu fruit glycosides. The purity of the obtained tofu glucoside is greater than 95%, and the yield of the tofu glucoside is 1.57% of that of the sow fruit seeds.

Among them, coix seed lactone is extracted and purified according to the following method, and undergoes the following steps:

Crush coix seed, pass it through a 100-mesh sieve, and then conduct supercritical carbon dioxide extraction for 2 hours under the conditions of temperature 35°C, pressure 40MPa, CO ₂ flow rate 10kg/h, and use ethanol with a volume concentration of 85% to dissolve and extract substance, and then the ethanol was removed by rotary evaporation to obtain yellow oil, which was coix seed lactone. The obtained coix seed lactone extraction yield was 5.9%.

The above-mentioned allicin, tofu glucoside and coix seed lactone extracts are fully mixed in proportion, and then pharmaceutically acceptable auxiliary materials are added to prepare any commonly used granules, pills or tablets.

Example 3

The pharmaceutical composition for treating neurasthenia and preventing senile dementia consists of the following components in mass percentage:

Garlic Nervous Acid 10%,

Tofu glucoside 1%,

Coixenolide 1%,

Excipients 88%,

The auxiliary materials are dextrin, lactose, sodium carboxymethyl starch, microcrystalline cellulose, starch, and hydroxypropyl cellulose.

Wherein the garlic fruit nerve acid is extracted and purified according to the following steps:

(1) Remove the shell and epidermis of the garlic fruit seeds, according to the solid-to-liquid ratio g/mL of 8:5, mash with phosphate buffer solution with a concentration of 20mM and pH=7.0 at 4°C, and then ultrasonically pulverize 30min, extract overnight at 4°C, then filter with 4 layers of gauze, then centrifuge at 8000 r/min at 4°C for 30min, repeat twice, remove the supernatant to obtain the residue; The buffer solution is prepared according to the following operations: mix the disodium hydrogen phosphate solution with a concentration of 0.2M and the sodium dihydrogen phosphate solution with a concentration of 0.2M at a volume ratio of 46:68 to obtain a phosphate buffer solution with pH=7.0; the supernatant The solution was purified and separated to obtain protein precipitation;

(2) Add petroleum ether to the residue obtained in step (1) according to the solid-to-liquid ratio g/mL=1:5 for stirring, then let it stand for stratification, collect the filtrate after filtration, and soak the filter residue 3 times with the filtrate repeatedly, each Filter after soaking for 60 minutes for the first time, combine the filtrates, and then carry out rotary evaporation at 50°C to remove petroleum ether to obtain light yellow garlic fruit oil;

(3) Weigh the silica gel G with a particle size of 200 mesh according to the mass ratio of the sample amount: the amount of silica gel = 1:10, first add petroleum ether to dissolve the silica gel G according to the solid-liquid ratio g/mL=1:10, and then stir for 30 minutes. Pour it into the chromatographic column. After the silica gel chromatographic column does not contain air bubbles and the sedimentation is stable, the garlic fruit oil obtained in step (2) is dissolved in petroleum ether according to the solid-to-liquid ratio g/mL=1:8, and then loaded into The upper layer of the silica gel chromatography column is gradiently eluted with petroleum ether-ethyl acetate elution system, according to the fraction of petroleum ether dissolved with garlic fruit oil:ethyl acetate=30:1 (V/V), at below 40°C Concentrate the fractions under reduced pressure to obtain colorless blocky crystals, which are allicin.

The nervonic acid standard and the colorless crystal were spotted on the same thin-layer plate by thin-layer chromatography, developed, and compared. As a result, the colorless crystal and the nervonic acid standard showed the same color at the same position on the same thin-layer plate. Purple spots, and the same color spots and the same Rf value in three different unfolding systems, prove that the colorless crystal is nervonic acid. The content of nervous acid obtained by this method is greater than 95%. The yield of nervonic acid is 10.819% of that of garlic fruit oil.

Wherein the tofu glucoside is extracted and purified according to the following method, and undergoes the following steps:

(1) Crush the bean curd seeds, pass through a 100-mesh sieve, then add ethanol with a volume concentration of 50%, soak for 30 minutes at 30°C, then extract with ultrasonic waves for 30 minutes, filter, and concentrate the filtrate by rotary evaporation at 55°C to recover ethanol. Obtain the ethanol extract of sow fruit;

(2) Add pure water to the tofu fruit ethanol extract obtained in step (1) according to the solid-to-liquid ratio g/mL of 1:5 to dissolve, apply macroporous resin D101 type, and first elute with pure water to remove polysaccharides, proteins, etc. Water-soluble impurities, then elute 4 retention volumes with ethanol with a volume concentration of 30%, collect the ethanol effluent with a volume concentration of 50% of the macroporous resin, concentrate under reduced pressure, and then add 2 times the volume of the concentrate to a concentration of 90 % ethanol, stand at 4°C for 12 hours to precipitate crystals, and then filter out the crystals to obtain pure tofu fruit glycosides. The purity of the obtained tofu glucoside is greater than 95%, and the yield of the tofu glucoside is 1.13% of that of the sow fruit seeds.

Among them, coix seed lactone is extracted and purified according to the following method, and undergoes the following steps:

Crush coix seed, pass it through a 100-mesh sieve, then extract coix seed powder with supercritical carbon dioxide for 1.5 h at a temperature of 38°C, a pressure of 38 MPa, and a _CO flow rate of 20 kg/h, and dissolve it in ethanol with a volume concentration of 95%. Extract the substance, and then remove the ethanol by rotary evaporation to obtain a yellow oil, which is coix seed lactone. The extraction yield of coix seed lactone was 5.77%.

The above-mentioned allicin, tofu glucoside and coix seed lactone extracts are fully mixed in proportion, and then pharmaceutically acceptable auxiliary materials are added to prepare any commonly used oral liquid.

Get the pharmaceutical composition of above-mentioned embodiment gained treatment neurasthenia and prevention senile dementia, be used for following clinical treatment:

1. Subjects of treatment

30 cases of neurasthenia patients; 30 cases of senile dementia patients; 50 cases of depression patients.

The above-mentioned cases were randomly divided into treatment group and control group, 55 cases in each group.

Two, usage

The treatment group took the pharmaceutical composition for treating neurasthenia and preventing senile dementia obtained in the above examples, 2 g each time, once a day, for 30 consecutive days for observation.

The control group took commercially available Shenshuaikang capsules, several times a day, 5 capsules each time, twice a day, for 30 consecutive days for observation.

3. Results

The clinical therapeutic effects of the pharmaceutical composition for treating neurasthenia and preventing senile dementia provided by the present invention: the effective rate reaches 100%, and the effective rate reaches 83% for the symptoms of neurasthenia patients; the effective rate for the symptoms of senile dementia patients reaches 78%; The effective rate of symptoms in patients with depression is as high as 90%.

The control group has an effective rate of 75% for the symptoms of neurasthenia; 65% for the symptoms of Alzheimer's patients; and 43% for the symptoms of depression patients.

4. Conclusion

The pharmaceutical composition for treating neurasthenia and preventing senile dementia provided by the present invention is a composition composed of allicin, tofu glucoside, coix seed lactone extract, etc., and has significant anti-senile dementia, neurasthenia and anti-tumor effects. Curative effect, its effect is better than that of the control group.

The pharmaceutical composition for treating neurasthenia and preventing senile dementia is completely extracted and purified from plants, the raw materials are easy to obtain, and has the advantages of good curative effect, safe application, and small toxic and side effects. The prepared product is stable in quality and easy to use, and is more suitable for Clinical medication, without any side effects, the curative effect is remarkable.

Claims (4)

1. A pharmaceutical composition for treating neurasthenia and preventing senile dementia, characterized in that it is made up of the following components by mass percentage: Garlic Neural Acid 1～10%, Tofu fruit glycosides 1~10%, Coix seed lactone 1～10%, The balance is auxiliary materials; The excipients include conventional dextrin, lactose, sodium carboxymethyl starch, microcrystalline cellulose, starch, and hydroxypropyl cellulose in the pharmaceutical field; Wherein, the garlic nervous acid is extracted and purified according to the following steps: (1) Remove the shell and epidermis of the garlic seeds, and pound them with a phosphate buffer solution with a concentration of 20mM and a pH of 6.0 to 7.0 at 3 to 4°C according to the solid-to-liquid ratio g/mL of 1 to 10:5 to 50. After being crushed, it was subjected to ultrasonic crushing treatment for 30-50 minutes, extracted at 3-4°C overnight, then filtered, and then centrifuged at 5000-8000 r/min at 3-4°C for 30-40 minutes. Repeat twice, remove the supernatant to get the residue; (2) Add petroleum ether to the residue obtained in step (1) according to the solid-to-liquid ratio g/mL=1～5:5～20 for stirring, then let it stand for stratification, collect the filtrate after filtration, and soak the filter residue repeatedly with the filtrate 2 to 3 times, soaking for 30 to 60 minutes each time, then filtering, combining the filtrates, and performing rotary evaporation at 40 to 50°C to remove petroleum ether to obtain garlic fruit oil; (3) Weigh the silica gel G according to the mass ratio of the sample amount: the amount of silica gel = 1 ~ 10: 10 ~ 100, first add petroleum ether to dissolve the silica gel G according to the solid-liquid ratio g/mL = 1 ~ 5: 2 ~ 10, and then After stirring for 20-30 minutes, pour it into the chromatographic column. After the silica gel chromatographic column does not contain air bubbles and the sedimentation is stable, the garlic fruit oil obtained in step (2) is mixed according to the solid-liquid ratio g/mL=1-5:1 ~10 After adding petroleum ether to dissolve, put it into the upper layer of the silica gel chromatography column, and use petroleum ether-ethyl acetate elution system to carry out gradient elution, according to petroleum ether dissolved with garlic fruit oil:ethyl acetate=50:1～30 : 1 (V/V) fraction, concentrate the fraction under reduced pressure at 40-50°C to obtain colorless massive crystals, which is allicin nervine acid; The tofu fruit glycosides are extracted and purified according to the following method, through the following steps: (1) Crush the bean curd seeds, pass through a 100-mesh sieve, then add ethanol with a volume concentration of 50-70%, cold-soak at 20-30°C for 30-60 minutes, then ultrasonically extract for 30-60 minutes, filter, and the filtrate is at 45-55 Carry out rotary evaporation and concentration at ℃, recover ethanol, and obtain sow fruit ethanol extract; (2) Add pure water to the tofu fruit ethanol extract obtained in step (1) according to the solid-to-liquid ratio g/mL of 1-5:5-25 to dissolve, apply macroporous resin D101 type, and first elute with pure water to remove Water-soluble impurities such as polysaccharides and proteins, and then elute 4 to 5 retention volumes with ethanol with a volume concentration of 30 to 50%, collect the ethanol effluent with a volume concentration of 30 to 50% of the macroporous resin, concentrate under reduced pressure, and then Add ethanol with a concentration of 90-95% 2-3 times the volume of the concentrate, let stand at 3-4°C for 12-24 hours to precipitate crystals, and then filter out the crystals to obtain pure tofu fruit glycosides; The coix seed lactone is extracted and purified according to the following method; Crush coix seed, pass through a 100-mesh sieve, and then extract coix seed powder with supercritical carbon dioxide for 1-2 hours at a temperature of 35-40°C, a pressure of 37-40 MPa, and a _CO flow rate of 10-20 kg/h. Dissolve the extract in ethanol with a concentration of 70-95%, and then remove the ethanol by rotary evaporation to obtain yellow oil, which is coix seed lactone. 2. The pharmaceutical composition according to claim 1, characterized in that: the phosphate buffer solution in the step (1) is prepared according to the following operations: disodium hydrogen phosphate solution with a concentration of 0.2M and phosphoric acid with a concentration of 0.2M The sodium dihydrogen solution is mixed according to the volume ratio of 12.3-61:87.7-39 to obtain a phosphate buffer solution with a pH of 6.0-7.0. 3. The pharmaceutical composition according to claim 1, characterized in that: the supernatant of the step (1) is subjected to subsequent purification and separation to obtain protein precipitation. 4. The pharmaceutical composition according to claim 1, characterized in that: the particle size of the silica gel G in the step (3) is 200-300 mesh.