KR20040023196A - Phamaceutical composition for treating Alzheimer comprising, as main ingredients, Aurantii Nobilis Pericarpium, Hoelen, Ligusticum acutilobum, Uncariae Ramulus et Uncus, Acorus gramineus, Atractyodis Rhizoma, Angelica dahurica, Zizyphi Spinosi Semen, Rehmanniae Radix Preparata, Cornus officinalis and Polygala tenuifolia - Google Patents

Abstract

PURPOSE: Provided is a medicinal composition which exerts excellent treatment effect in treating Alzheimer's disease, multi-infarct dementia, a combination type of Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyroidism, and alcoholic dementia. CONSTITUTION: The medicinal composition comprises extracts of Aurantii nobilis pericarpium, Hoelen, Ligusticum acutilobum, Uncariae ramulus et uncus, Acorus gramineus, Atractyodis rhizoma, Angelica dahurica, Zizyphi spinosi semen, Rehmanniae radix preparata, Cornus officinalis and Polygala tenuifolia as main components, and extracts of Cyperus rotundus, Pinellia ternate, Cnidium officinale, Paeonia lactiflora, Platycodi radix, Ponciri fructus, keel, Ostreae concha, Inula helenium, Biotae semen, Cinnamomum loureirii, Disocorea batatas, Chinese matrimony vine, snake's beard, Glycyrrhizae radix and Schizandrae fructus as subsidiary components. The extraction is made by a solvent selected from the group consisting of water, low chain alcohol, ethyl acetate, aromatic hydrocarbon, and chlorinated hydrocarbon.

Description

Pharmaceutical composition for treating Alzheimer comprising, as main ingredients, Aurantii Nobilis Pericarpium, Hoelen, Ligusticum acutilobum, Uncariae Ramulus et Uncus, Acorus gramineus, Atractyodis Rhizoma, Angelica dahurica, Zizyphi Spinosi Semen, Rehmanniae Radix Preparata, Cornus officinalis and Polygala tenuifolia}

The present invention relates to the dermis (Aurantii Nobilis Pericarpium), Hoelen, Ligusticum acutilobum, Uncariae Ramulus et Uncus, Acorus gramineus, Atractyodis Rhizoma, White paper (Angelica dahurica), Sanjoin (Z) Alzheimer's disease, vascular disease (multi-infarct dementia: Multi-infarct), containing Spinosi Semen, Rehmanniae Radix Preparata, Cornus officinalis, and Polygala tenuifolia, or extracts derived from their organic or inorganic solvents. infarct dementia), a mixture of Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, and alcoholic dementia.

Dementia is a syndrome caused by brain disorders that is usually chronic and progressive, and is a multiple disorder of brain function that includes memory, thinking, coping, computing, learning, language, and judgment. The normal person is unable to maintain normal family life, social life and various interpersonal relationships due to gradual memory impairment and other loss of intellectual ability due to the disease. Loss of sensation, impaired judgment, personality changes, learning disabilities, and loss of language skills make the patient unable to care for himself. It is also a frightening senile disease that continues from 3 to 20 years. The incidence rate is similar for both men and women, which is the fourth leading cause of death for the elderly. The incidence of dementia is barely noticeable, and in Korea, more than 8% of the elderly aged 65 or older last year are expected to exceed 10% by 2020. As aging progresses, dementia patients are bound to increase. It is estimated that about 10% of elderly people have dementia.

Since it has recently been found that beta amyloid protein is one of the important causative factors for the development of Alzheimer's disease, molecular biological research on beta amyloid is expected to contribute the most to the disease. An important lesion of senile dementia is the formation of neurofibrillary tangles and neuritic plaques by the deposition of beta amyloid proteins consisting of between 39 and 43 amino acids. As described above, it is mainly deposited on cholinergic nuclei and hippocampal structures of the basal forebrain. The beta amyloid protein is formed by cleavage from a large molecular weight of the precursor protein (amyloid precursor protein 695, 751, 770: APP 695, 751, 770). However, at present, the amyloid precursor protein is cleaved by an enzyme and by a metabolic process to produce beta A4 protein. Therefore, efforts to express protease that promotes the formation of beta amyloid protein and genes of this protease are the core of the current research, and the development of drugs that inhibit the expression of this protease enzyme protein or protease gene is the future drug. It is the biggest goal in development. In addition, the method of activating the amyloid degrading enzyme (ADE) in the normal can be an important development goal.

There is also a lot of debate about the intracellular function of the amyloid precursor proteins attached to the cell membrane and the function of the beta A4 protein that is cut off from the amyloid precursor protein and deposited on the tissue. Therefore, the challenges required to advance such research and development will be the expression, mass production, and purification of amyloid precursor protein and beta A4 amyloid protein using the latest genetic engineering techniques. In addition to producing antibodies using the recombinant protein thus obtained, the in vivo function of amyloid precursor proteins and the role of beta amyloid protein in the pathogenesis of Alzheimer's disease can be precisely identified. We can study how metabolism works. Recently, various causes of cerebral neuropathy, ischemic nerve injury, and central nervous system disorder have been reported to be directly related to the generation of reactive oxygen species and the regulation of degradation mechanisms. Induced by the dementia inducer due to the generation of reactive oxygen species and the brain cell membrane is damaged and various brain neurological disorders are found to occur. (FIG. 1).

Recently, research on the neurotransmitter system has been conducted, and a rational approach to treatment has been sought. Attempts have been made to improve cholinergic function, and glucose and oxygen metabolizers have been tried. However, the possible pharmacological approaches already discussed have at best a temporary relief. Clinical results of Cognex, Aricept, and Memantine, a new drug for vascular dementia, recently developed by Merz + Co. Was released. The clinical study was conducted in a randomized, placebo-comparison test, and the results of the study were presented at the 6th International Stockholm / Springfield Alzheimer's Disease Development Symposium on April 5-8 in Stockholm, Sweden. Published by Symposium on Advances in Alzheimer Therapy, the company developed Novatis in the US for clinical applications in Europe but has not been approved by the FDA. Exelon, developed by Novartis, is widely used in Europe, but has not yet been approved by the FDA. The drug enhances memory and cognition by inhibiting the action of the enzyme acetylcholine, a neurotransmitter in the brain. In the United States, clinical trials of 941 patients with early or mid-term Alzheimer's disease reported a significant improvement in symptoms in 26%, with 82% showing no worsening or improvement in symptoms. Compared to Cognex, the ingredient Tacrine has no liver toxicity and is easier to take. As an acetylcholinease inhibitor, the effect is similar, but due to liver toxicity, patients often had to undergo liver function tests and blood tests. Because it is taken four times a day, it was very difficult for the patient to take the medicine. Aricept is taken once a day and exelon is taken twice a day. The development trend of the dementia treatment in Korea is that dementia is caused by not only the 100 amino acid masses located at the end of the carboxyl group (end C), but also the nerve cells are directly destroyed and the glial cells are stimulated to cause inflammation. According to Yoo Heon Hee of the Seoul National University School of Medicine, "Currently, six to seven toxic proteins are known to cause dementia by acting on the brain, but C protein is considered to be the most fundamental cause of dementia." It is said to be in development as dihydroebodiamine and has begun Phase I clinical trials from this year after the preclinical trial was completed at the end of last year. The dementia developed by Wonkwang University's team reported better results than Cognex and Aricept, which improve blood circulation and memory.

Alzheimer's disease has been reported to treat Alzheimer's disease by inhibiting enzymes that break down and deplete acetylcholine. Three representative products are Aricept (component name Donepezil: Korea Ezai), Exelon (component name Rivastigmine: Novartis Korea), and reminil (component name Galantamine: Jansen Korea). Three years ago, tacrine, piracetam, and idebenone, which activate the brain's mitochondria, were used a lot, but there are many side effects and insufficient efficacy, which is rarely used now. Some of the treatments for Alzheimer's disease are effective against vascular dementia and alleviate mental and behavioral problems. However, it is reported that cognitive deterioration cannot be restored and can be maintained for 2 to 5 years when used in patients with mild or moderate symptoms.

According to a study conducted last year in Canada and the United Kingdom, Aricept is also effective for patients with moderate or severe Alzheimer's disease, and has been shown to be superior to excellon and reminil in terms of efficacy and side effects. Aricept should be taken only once a day. However, execlone and reminil should be gradually increased to a therapeutic effect and taken two to three times a day. These drugs have side effects such as hot flashes, indigestion, nausea, vomiting, and diarrhea, most of which are tolerable and recover over time. At high doses, anorexia, bradycardia, muscle weakness, and dizziness may appear, but they disappear on their own, but they may give up due to difficulty taking the medicine. Drugs that are expected to alleviate symptoms of Alzheimer's disease include tocopherol (vitamin E), ginkgo biloba extract, and estrogen. Vascular dementia is a dementia caused by cumulative damage and blockage of cerebrovascular disease due to hypertension, diabetes, hyperlipidemia, heart disease, smoking, obesity and lack of exercise. Therefore, the first thing to treat is the cause. Next, to prevent clogging due to narrowing of blood vessels, drugs such as aspirin or ticlopidine may be used to prevent the formation of blood clots, or warfarin, which inhibits the sticking of blood.

Although not commonly used as a treatment for dementia, seregiline (a treatment for Parkinson's disease and mild dementia) and nisergoline (a treatment for cerebral infarction and vascular dementia) are also used as treatments for dementia. In addition, dementia patients have symptoms such as anxiety, anxiety, depression, delusions, hallucinations, violence, and insomnia in addition to cognitive impairment, so they often take antidepressants and anti-anxiety drugs together. The need for the emergence of drugs that are more effective and safer in the prescription without side effects of toxicity.

As other treatments for dementia caused by reactive oxygen species have not been clearly identified, in order to have an effect as a treatment for dementia, the herbal medicines were first searched for the antioxidant function without brain toxicity. Results The present inventors have developed a drug useful in the treatment of dementia with excellent antioxidant function while suppressing app expression caused by reactive oxygen species from a medicinal herb derived from herbal medicines. The present invention is to provide a drug useful in the treatment of dementia excellent in antioxidant function while suppressing the app expression by the reactive oxygen species as described above.

Figure 1 shows the process of generating the causative agent of the dementia pathogenesis.

Figure 2 shows a method for recovering the dry weight of the pharmaceutical composition according to the extraction process of the effective drug.

Figure 3 shows the SOD enzyme activity of the brain tissue by CHU-1.

Figure 4 is a graph showing the xanthine oxidase enzyme activity of the brain tissue by CHU-1.

Figure 5 shows the inhibitory effect of peroxidation substrate of brain tissue by CHU-1.

Figure 6 shows the activity of DPPH through the effective drug CHU-1.

7 is a graph showing the effect on the clearance of NO through the effective drug CHU-1

8 is analyzed by RT-PCR whether the effective drug CHU-1 inhibits the transcriptional expression of iNOS and COX-II in brain tissue by free radicals.

9 is analyzed by western blot whether the effective drug CHU-1 inhibits the expression of iNOS and COX-II in brain tissue by free radicals.

The present inventors have conducted a long study on herbal medicines that can be used for dementia and related brain diseases. As a result, the dermis (Aurantii Nobilis Pericarpium), Hoelen, Ligusticum acutilobum, Unguiae Ramulus et Uncus, which are present in the extract X (hereinafter referred to as "CHU-1") extracted in Example 1 of the present invention. ) Powder or water from Acorus gramineus, Atractyodis Rhizoma, Angelica dahurica, Zizyphi Spinosi Semen, Rehmanniae Radix Preparata, Cornus officinalis, Polygala tenuifolia , Extracted from lower alcohols, ethyl acetate, aromatic hydrocarbons, chlorinated hydrocarbons, and selected as a main ingredient, which includes Cyperus rotundus, Pinellia ternata, Cnidium officinale, and Peony lactiflora, Platycodi Radix, Ponciri Fructus, keel, Ostreae Concha, Inula helenium, Biotae Semen, Schizandra chinensis, Cinnamomum loureirii, Potion (Disocorea batat as, at least one supplementary drug selected from chinese matrimony vine, snake's beard, licorice (GLYCYRRHIZAE RADIX), or Schizandrarae FRUCTUS or its water, lower alcohols, ethyl acetate, aromatic hydrocarbons or chlorinated hydrocarbons A pharmaceutical composition composed of X extracted with selected solvents, which is composed of Alzheimer's disease, vascular disease (Multi-infarct dementia), Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, alcoholic dementia The present invention was completed by the discovery that it shows an excellent effect on Alzheimer's treatment. Therefore, the development of drugs that inhibit the dementia CT105 is currently in progress. And current research suggests that these drugs may serve as a useful treatment for future dementia.

The present invention is 50 to 500 parts by weight of dermis (Aurantii Nobilis Pericarpium), 50 to 500 parts by weight of Hoelen, 50 to 500 parts by weight of Ligusticum acutilobum, 50 to 500 parts by weight of Uncariae Ramulus et Uncus, Seokchangpo (Acorus gramineus) 50 to 500 parts by weight, 50 to 500 parts by weight of Atractyodis Rhizoma, 50 to 500 parts by weight of Angelica dahurica, 50 to 500 parts by weight of Zizyphi Spinosi Semen, Rehmanniae Radix Preparata 50 to 500 parts by weight, Cornus officinalis 50 to 500 parts by weight, raw paper (Polygala tenuifolia) 50 to 500 parts by weight or its extract as a main ingredient, if necessary here 50 to 500 parts by weight of Cyperus rotundus, 50 to 500 parts by weight of Pinellia ternata, 50 to 500 parts by weight of Cnidium officinale, 50 to 500 parts by weight of Paeonia lactiflora, 50 to 500 parts by weight of Platyodi Radix, and Ponciri Fructus 50 To 500 parts by weight, keel 50 to 500 parts by weight, 50 to 500 parts by weight of Ostreae Concha, 50 to 500 parts by weight of Inula helenium, 50 to 500 parts by weight of Biotae Semen, 50 to 500 parts by weight of Schizandra chinensis, 50 to 500 parts by weight of Cinnamomum loureirii, 50 to 500 parts by weight of Dosocorea batatas, 50 to 500 parts by weight of chinese matrimony vine, 50 to 500 parts by weight of snake's beard, GLYCYRRHIZAE RADIX 50 to 500 parts by weight, SCHIZANDRAE FRUCTUS 50 to 500 parts by weight of one or more auxiliary drugs selected from or a solvent selected from water, lower alcohol, ethyl acetate, aromatic hydrocarbons or chlorinated hydrocarbons is added to the molecule In biological experiments, the results provide a remarkable effect of inhibiting dementia progression.

Another object of the present invention is 50 to 500 parts by weight of dermis (Aurantii Nobilis Pericarpium), 50 to 500 parts by weight of Hoelen, 50 to 500 parts by weight of Ligusticum acutilobum, Uncariae Ramulus et Uncus 50 to 500 parts by weight Part, 50 to 500 parts by weight of Acorus gramineus, 50 to 500 parts by weight of Atractyodis Rhizoma, 50 to 500 parts by weight of Angelica dahurica, 50 to 500 parts by weight of Zizyphi Spinosi Semen, Rehmanniae Radix Preparata) 50 to 500 parts by weight, Cornus officinalis 50 to 500 parts by weight, raw paper (Polygala tenuifolia) 50 to 500 parts by weight or its extract, if necessary, here are 50 to 50 per centus (Cyperus rotundus) 500 parts by weight, 50 to 500 parts by weight of Pinellia ternata, 50 to 500 parts by weight of Cnidium officinale, 50 to 500 parts by weight of Paeonia lactiflora, 50 to 500 parts by weight of Platyodi Radix Ponciri Fructus) 50 to 500 weight Boil, keel 50 to 500 parts by weight, Ostreae Concha 50 to 500 parts by weight, Inula helenium 50 to 500 parts by weight, Biotae Semen 50 to 500 parts by weight, Schizandra chinensis 50 to 500 parts by weight, 50 to 500 parts by weight of Cinnamomum loureirii, 50 to 500 parts by weight of Dosocorea batatas, 50 to 500 parts by weight of chinese matrimony vine, 50 to 500 parts by weight of snake's beard , GLYCYRRHIZAE RADIX 50 to 500 parts by weight, SCHIZANDRAE FRUCTUS 50 to 500 parts by weight of one or more auxiliary herbs or water, lower alcohol, ethyl acetate, aromatic hydrocarbons or chlorinated hydrocarbons extracted with a solvent selected from X It is to provide a pharmaceutical formulation prepared by a conventional method for preparing a pharmaceutical formulation by adding a pharmaceutically acceptable excipient to.

Each herbal ingredient may be powdered, or each may be extracted alone to obtain an extract, and these extracts may be mixed, or the necessary herbal ingredients may be combined and extracted together.

The present invention is described in more detail by the following examples and experimental examples, which do not limit the scope of the present invention.

Example 1

1.100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma 90 g of Angelica dahurica, 90 g of Zizyphi Spinosi Semen, 90 g of Rehmanniae Radix Preparata, 90 g of Cornus officinalis, and 80 g of Polygala tenuifolia are added in a volume of 3 to 5 times distilled or purified water. After heating slowly for 12 hours under 75 ° C. conditions, the mixture was filtered and evaporated to a final volume of 500 ml, and then concentrated under reduced pressure to 200 ml, and then frozen to obtain 75 g of lyophilized weight ( Hereinafter referred to as CHU-1). (Figure 2)

2. Extracted by the above method, each extract about 10 times the amount of each herbal medicine with about 3000ml of distilled water, then the extracts are combined and treated by the above method to obtain about 750g of lyophilisate.

Example 2

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, 100 g of white paper (Angeica) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia with 2 to 5 times the volume of purified water with 50% (v / v) ethanol Heated to 12 hours under 60 ° C., filtered, evaporated to a final volume of 500 ml, concentrated under reduced pressure to 200 ml, and then frozen and lyophilized to about 75 g. Gained weight

Example 3

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia are added to 70% (v / v) ethanol in purified water and 75 ℃ After heating slowly for 12 hours under the conditions, the resultant was filtered, the resultant was evaporated to a final volume of 500 ml, concentrated under reduced pressure to 200 ml, and then frozen to obtain a lyophilized X.

Example 4

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, Zizyphi Spinosi Semen 90g, Rehmanniae Radix Preparata 90g, Cornus officinalis 90g and Polygala tenuifolia 80g Cyperus rotundus 100g, Pinellia ternataale 100ni of cinin ) 100 g was processed by the method of Example 1-1 to obtain a lyophilized product.

Example 5

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia, 50g of Paeonia lactiflora, 50g of Platyodi Radix Fructus) 50g was processed by the method of Example 1-1 to obtain a lyophilisate.

Example 6

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis, 80g of Polygala tenuifolia, 50g of keel, 50g of Ostreae Concha, 50g of Inula helenium 50 g, Biotae Semen 50 g, and Cinnamomum loureirii 50 g were treated in the same manner as in Example 1-1 to obtain a lyophilized product.

Example 7

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia, 50g of Disocorea batatas, 50g of Chinese matrimony vine 50g of snake's beard), 50g of licorice (GLYCYRRHIZAE RADIX), and 50g of Schizandrae FRUCTUS were treated by the method of Example 1-1 to obtain a lyophilized product.

Example 8

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90 g of Zizyphi Spinosi Semen, 90 g of Rehmanniae Radix Preparata, 90 g of Cornus officinalis, 80 g of Polygala tenuifolia, 100 g of Cyperus rotundus, 100 g of Pinellia ternatain Cini ) 50g of Disocorea batatas, 50g of goji (chinese matrimony vine), 50g of snake's beard, 50g of licorice (GLYCYRRHIZAE RADIX), 50g of Schizandrarae FRUCTUS by the method of Example 1-1 Got.

Example 9

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) ) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia, 100g of Cyperus rotundus, 100g of Pinellia ternataC 50 g officinale 100 g of isocorea batatas, 50 g of goji (chinese matrimony vine), 50 g of snake's beard, 50 g of GLYCYRRHIZAE RADIX, 50 g of Schizandrarae FRUCTUS were frozen by the method of Example 1-1. A dried product was obtained.

Example 10

50 g of dried dermis (Aurantii Nobilis Pericarpium), 50 g of Hoelen, 50 g of Ligusticum acutilobum, 50 g of Uncariae Ramulus et Uncus, 50 g of Agrarus gramineus, 50 g of Atractyodis Rhizoma, dagica (Angelica) ) 50g, 50g of Zizyphi Spinosi Semen, 50g of Rehmanniae Radix Preparata, 50g of Cornus officinalis and 50g of Polygala tenuifolia, 50g of Cyperus rotundus, 50g of Pinellia ternataCnig 50 g officinale, 50 g of Disocorea batatas, 50 g of Chinese matrimony vine, 50 g of snake's beard, 50 g of GLYCYRRHIZAE RADIX, and 50 g of Schizandrarae FRUCTUS were treated by the method of Example 1-1. Lyophilized was obtained.

Example 11

100 g of dried dermis (Aurantii Nobilis Pericarpium), 100 g of Hoelen, 90 g of Ligusticum acutilobum, 90 g of Uncariae Ramulus et Uncus, 90 g of Agrarus gramineus, 90 g of Atractyodis Rhizoma, dagica (Angelica) 90g, 90g of Zizyphi Spinosi Semen, 90g of Rehmanniae Radix Preparata, 90g of Cornus officinalis and 80g of Polygala tenuifolia were powdered to obtain a powder.

Experimental Example 1

To investigate the antioxidant function of the obtained CHU-1, 6 males of 200 g of SD rats, which were experimental animals, were anesthetized with ketamin, followed by an incision in the brain. pH 7.5) was added and triturated with a homogenizer (Heidolph RZR 2021) at about 4 ° C. The grinding homogenate was centrifuged at 600 xg for 10 minutes to remove the nucleus and ungrind. Then, the supernatant was ultracentrifuged at 100,000 xg for 1 hour to obtain cytosol fraction, which was used as an enzyme source. All the above operations were performed at 0-4 degreeC. Superoxide is produced by intracellular NADP oxidase, which is converted to hydrogen peroxide (H ₂ O ₂ ) by superoxide dismutase (SOD) (Bodel, 1996), and H ₂ O ₂ is catalase (CAT). It becomes H ₂ O which is harmless. However, large amounts of H ₂ O _{2 form} iron toxins as catalysts to form toxic substances such as hydroxyl radicals (· OH) (Miles et al., 1996). These toxic substances are called reactive oxygen intermediates (ROIs) and are placed in a test tube with 50 mM potassium phosphate buffer (containing 0.1 mM EDTA, pH = 7.8), 17 μl of distilled water, 17 μl of enzyme sample, and 17 μl of 5 μM sodium xanthine. Absorbance was measured at 25 ° C. and 550 nm in a thermostat. SOD was analyzed three times using absorbance based on Sen et al.'S (1994) analysis method. Excessive free radicals oxidize unsaturated fatty acids or lipids in our body and slowly kill brain cells without being excreted in the kidneys. Makes peroxidated lipid peroxide. In addition, free radicals oxidize cholesterol in the bloodstream and make lipid peroxide, which continuously attaches to blood vessel walls. Lipid peroxide is a source of adult disease, a highly toxic and deadly brain that kills brain cells. Most of the cells are made of lipids, which are easy to oxidize, and are easily oxidized by the action of strong free radicals, resulting in lipid peroxide that kills or damages brain cells and various tissues in our body. The control group was 21.51 ± 0.75% relative unit, and this CHU-1 showed about 60% of the enzyme activity compared to the control group. Therefore, this drug is expected to exhibit SOD-like function in brain cells. It was shown to increase the enzyme activity by about 60% compared to the control at a concentration of 0.05 mg / ml (Fig. 3).

Experimental Example 2

Xanthine oxidase, a type of free radical synthase, plays an important role in catalyzing the biochemical reaction by converting from xanthine dehydrogenase during radical production. The method of stripe is used to measure the enzyme activity after oral administration of CHU-1 in rats. According to the method, 60 μM of substrate xanthine and 0.2 ml of enzyme source were added to 0.1 M potssium phosphate buffer (pH 7.5), and then reacted at 37 ° C. for 5 minutes. 20% trichloroacetic acid was added to remove proteins and centrifuged. The activity of the enzyme was determined by measuring the change in absorbance at 292 nm of the uric acid produced. Xanthine oxidase is a free radical synthase that plays an important role in catalyzing the biochemical reaction in the radical production process. As a result of measuring enzyme activity in the reaction solution for measuring xanthine oxidase activity using cytosol of rat brain tissue as an enzyme source and xanthine substrate, the enzyme activity was inhibited by about 50% compared to the control at 0.05 mg / ml. (FIG. 4). In comparison with the normal group, the inhibitory activity was about 13% less than that of CHU-1.

Experimental Example 3

Lipid peroxide content in vitro was determined by adding 0.1% of brain tissue homogenate to 8 ml sodium dodesyl sulfate, 20% acetic acid buffer (pH 3.5) and 0.8% 2-thiobarbituric acid according to Ohkawa et al. The reaction mixture was reacted for 1 hour at room temperature, cooled to room temperature, and the resulting red colored material was transferred to n-butanol: pyridine (15: 1, v / v) mixture and quantified by measuring the change in absorbance at a wavelength of 532 nm. Lipid peroxide content was expressed in nmol of malondialdehydee per gram of brain tissue. In the in vivo test, 50 μg / μl of CHU-1 was orally administered to the above-mentioned experimental animals for 30 days, the rats were treated, the brain tissues were extracted, and the amount of lipid peroxide was measured by the above method. Phospholipid unsaturated fatty acids cause peroxidation by various reactive oxygen species, and the lipid peroxide produced at this time is known to be a major cause of cell membrane damage including cerebral nerve cells by dementia and various reactive oxygen species. Therefore, inhibition of lipooxidation reaction is used as the most important indicator in the antioxidant activity of herbal medicines. The production of lipid peroxide was observed by adding brain tissue grinding fluid and CHU-1 in vitro under the experimental conditions of lipid peroxide production. As a result, CHU-1 showed a tendency to significantly inhibit the production of lipid peroxide (FIG. 5). . In particular, the amount of MDA produced at the concentration of 0.05 mg / ml was about 12.5 nmoles per gram of tissue, which suppressed the lipid peroxidation reaction by 45.4% compared to the control group and showed about 5.7 nmol difference compared to the normal group. .

Experimental Example 4

To measure the amount of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical, add 4 ml of 1.5x10 ^-5 Methanol solution of DPPH radical to 0.1 ml of CHU-1 according to Blosi's method, and react for 30 minutes at room temperature. Absorbance was measured and calculated. The scavenging activity of DPPH radical was calculated by the following equation.

Radical scavenging activity (%) =

{(OD control-ODsample) / ODcontrol} X 100

In the DPPH radical scavenging effect, which is used as another indicator of antioxidant activity, the maximum scavenging effect was 25 ㎍ / ml by CHU-1, which was about 4.7 ± 2.4. It can be seen that the effect of prooxidant is higher at higher concentrations. In contrast, the control group showed a relatively low scavenging effect on DPPHradical. CHU-1 was about 4 times higher in DPPH scavenging ability than in control group. Therefore, CHU-1 is analyzed to have an excellent effect of directly removing radicals (FIG. 6).

Experimental Example 5

In order to measure the NO production amount of CHU-1 obtained by the method of the present invention, subcutaneous injection of SNP at a concentration of 500 μM was added to 50 μg / μl of CHU-1, followed by oral administration of the drug for 30 days. To determine the ability to inhibit NO production, anesthetize the animals, and take the heart blood and dispense 100 μl of serum into 96 wells, and add Griess reagent (1% sulphanilamide, 0.1% naphlethyendiamine dihydrochloride, 4% phosphoric acod) The amount of NO production in the blood was measured by ELISA reader at 570 nm after treatment with 50 μl. The normal group was 4.0 ± 2.82μM and the control group was 98.2 ± 2.9μM, but the CHN-1 was 29.1 ± 2.5μM. It can be seen that the effect of inhibiting NO production about three times (FIG. 7).

Experimental Example 6

Expression of iNOS and COX-II Proteins by RT-PCR Analysis

CHU-1 obtained by the method of the present invention in order to determine the effect on the inhibition of iNOS, COX-II expression involved in inflammation and NO production, CHU after subcutaneous injection of SNP 100 μM concentration as described above -1 50 ㎍ / ㎕, and then oral administration of the drug for 30 days and then RNA zol solution was added according to the method described above to separate the nucleic acid and protein in the brain cells, centrifuged for 15 minutes at 15,000 rpm and the supernatant and isopropanol 200 μl was added to the mixture and allowed to stand in the refrigerator at 20 ° C. for 15 minutes, followed by centrifugation to separate total RNA, followed by RT-PCR, which was confirmed on 1% agarose gel. Induction of iNOS mRNA and COX-II mRNA expression during inflammation was strongly induced in the control group (lane 2), whereas it was strongly inhibited in the CHN-1 group (lane 3) ( 7) , COX-II mRNA expression was induced in the control group (lane 2). When neuronal cells induce neuronal cell death during CT105 expression, calpain expression was induced in the control group. The expression of MAPK involved in cell proliferation was strongly inhibited by CHU-1, and it was found that CHU-1 has an effect of inhibiting neuronal cell destruction or protecting neuronal cell death. In particular, the drug group was found to inhibit iNOS, COX-II expression and delay or inhibit apoptosis when sensitive to oxidative stress by active oxygen of neurons (FIG. 7).

Experimental Example 7

Western blot analysis from SNP-induced brain tissue

According to the present invention by CHU-1, the pharmaceutical composition isolated according to the present invention, to investigate whether the brain cells in the rat brain tissue involved in inflammation or scavenging of oxides, 1 g of brain tissue cells were taken and placed in liquid nitrogen. After trituration with Niger, lysis buffer (50 mM Tris-HCl pH 7.5, 1% (v / v) Triton X-100, 150 mM NaCl, 10% (v / v) glycerol, 2 mM dithiothreitol, 10 mM MgCl2) 30 μg of the extract was eluted on 10% polyacrylamide SDS gels (SDS-PAGE), and then transferred to an Immobilon-P membrane (Amersham) to confirm the expression of protein by ehanced chemilumenescence (ECL). (Santa Cruz, 1: 1,000 dilution) Label the primary antibody iNOS, COX-II, ERK 1/2, MAPK, wash with PBS, remove side reaction by adding blotting solution, and remove the secondary antibody, Horseradish Peroxidase-conjugate anti- Label goat IgG (HRP) antibody and incubate for 3 minutes with ECL blotting reagent. The chemiluminescence was confirmed by exposure to X-ray film for 30 minutes at 30 sec to confirm the color development. iNOS protein, which induces free radical and NO production by CT105 protein expression, and COX-II protein expression when inflammation was induced, While expression was strongly induced (lane 2), it was strongly inhibited in the CHU-1 group (lane 3) (FIG. 8), and in the control group in the expression of COX-II protein, the expression was induced. On the other hand (lane 2), on the other hand, when the neurons of neural tissues induce cell death upon expression of CT105, ERK1 / 2 (p42 / 44), which activates transcription factors during cell proliferation, is weakly reduced but not by CHU-1. There was a tendency to increase the level of expression similar to the normal group, and there was a slight difference in the expression of MAPK. These results suggest that CT105 expression induces the progression of cell death by decreasing the expression of ERK 1/2, but inhibits the destruction of neurons by CHU-1 and has a protective or protective effect on cell death. In particular, the drug group inhibits the expression of iNOS and COX-II, and exhibits delayed or inhibited apoptosis when sensitive to oxidative stress by active oxygen of neurons (FIG. 8). It is believed that this shows a tendency to strongly inhibit cell death.

As confirmed from the above experimental results, the novel composition of the present invention has an excellent antioxidative function while eliminating excellent reactive oxygen species. Therefore, the composition of the present invention can be usefully used as a pharmaceutical composition. The pharmaceutical composition of the present invention may vary depending on the sex, age, degree of disease, etc. of the patient, but if the composition of the present invention is used without extracting the composition of the present invention with water or an organic solvent, 1.0 g to 50 g per day is used. If used in the form of X, 10mg to 1000mg per day can be administered once to several times per day.

Compounds of the present invention are associated with stroke, stroke, cerebral infarction, hypertension, Alzheimer's disease, vascular disease (multi-infarct dementia), mixed Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, alcoholicity Since Alzheimer's dementia has a therapeutic effect, water extract has cognex, aricept, memantine, exelon, dihydroebodiamine, and reminil, which are already used as a medicament in the composition of the present invention. It further contains at least one drug selected from tacrine, piracetam, idebenone, tocopherol (vitamin E), ginkgo biloba extract, estrogen, aspirin, ticlopidine, warfarin, seregline, and nisergoline. It may also be used in combination with drugs such as excipients, adjuvants, diluents, tonicity agents, preservatives, suspending agents, dissolution aids and the like.

Combining the compound of the present invention with the above drug can reduce the normal dose of the existing drug, thus alleviating the problems with the existing drugs.

The compounds of the present invention may be mixed with excipients, adjuvants, analgesics, isotonic agents, preservatives, and other pharmaceutically acceptable adjuvants, which are commonly used pharmaceutically, and formulated into pharmaceutically acceptable formulations for pharmaceutical purposes. Suitable formulations may be prepared. Such pharmaceutical preparations may be formulated into injections, solutions, tablets, capsules, powders, syrups and the like.

Next, the present invention will be described in more detail as formulation examples.

Formulation Example 1

CHU-1 500mg

Appropriate sterile distilled water for injection

pH adjuster

Dissolve CHU-1 in distilled water for injection, adjust the pH to about 7.6 with a pH adjuster, make the whole to 2 ml, and fill a 2 ml ampoule and sterilize to prepare a medicinal needle injection.

Formulation Example 2

X 500mg of Example 2

Appropriate sterile distilled water for injection

pH adjuster

X 500 mg of Example 2 was dissolved in sterile distilled water for injection, adjusted to pH 7.2 with a pH adjuster, and the whole was made to 2 ml, and then filled in a 2 ml ampoule to prepare a medicinal needle injection.

Formulation Example 3

500 mg of water extract powder of Example 3

Lactose 100mg

Starch 100mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 4

X 500mg of Example 5

Lactose 100mg

Starch 50mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 5

X 500mg of Example 6

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the inside is filled with a polyethylene coated coated chloride and sealed to prepare a powder.

Formulation Example 6

X 500 mg of Example 8

Lactose 50mg

Starch 50mg

Talc 2mg

Magnesium stearate appropriate amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 7

X 500 mg of Example 9

Lactose 100mg

Starch 93mg

Talc 2mg

Magnesium stearate proper amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 8

X 5g of Example 1-1

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Add 100 ml of purified water

The above components are mixed according to a conventional method for preparing a liquid, and filled into 100 ml of brown bottle and sterilized to prepare a liquid.

Formulation Example 9

X 5g of Example 2

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Add 100 ml of purified water

The above components are mixed according to a conventional method for preparing a liquid, and filled into 100 ml of brown bottle and sterilized to prepare a liquid.

Formulation Example 10

X 500mg of Example 10

Cognex 10mg

Appropriate sterile distilled water for injection

pH adjuster

X 500 mg of Example 10 was dissolved in distilled water for injection, adjusted to pH about 7.6 with a pH adjuster, and then the total amount was 2 ml, followed by filling into a 2 ml ampoule and sterilizing to prepare an injection.

Formulation Example 11

X 500mg of Example 1-1

Aricept 10mg

Proper sterile distilled water for injection

pH adjuster

X 500 mg of Example 1-1 was dissolved in sterile distilled water for injection, adjusted to pH 7.2 with a pH adjuster, the total amount was 2 ml, and then filled in a 2 ml ampoule to prepare an injection.

Formulation Example 12

X 500mg of Example 1-1

Memantine 20mg

Lactose 100mg

Starch 100mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 13

X 500mg of Example 1-1

Excellon 50mg

Lactose 100mg

Starch 50mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 14

X 500mg of Example 2

Lactose 50mg

Starch 50mg

Talc 2mg

Magnesium stearate appropriate amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 15

X 1000mg of Example 1-1

Dihydroebodiamine 10mg

Lactose 50mg

Starch 50mg

Talc 2mg

Magnesium stearate appropriate amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 16

X 100mg of Example 6

Reminil 20mg

Lactose 100mg

Starch 93mg

Tackle 2mg

Magnesium stearate proper amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 17

X 1000mg of Example 7

Tacrine 1000mg

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Add 100 ml of purified water

The above components are mixed according to a conventional method for preparing a liquid, and filled into 100 ml of brown bottle and sterilized to prepare a liquid.

As described in detail above, the dermis (Aurantii NobilisPericarpium), Hoelen, Ligusticum acutilobum, Uncariae Ramulus et Uncus, Acorus gramineus, Acreyodis Rhizoma, White paper (Angelica dahurica) ), Zizyphi Spinosi Semen, Rehmanniae Radix Preparata, Cornus officinalis, Polygala tenuifolia powder, or a solvent selected from water, lower alcohol, ethyl acetate, aromatic hydrocarbons, chlorinated hydrocarbons, or mixtures thereof Extracted by the main ingredient, if necessary, Cyperus rotundus, Pinellia ternata, Cnidium officinale, Paeonia lactiflora, Platycodi Radix, Ponciri Fructus, Keel ), Ostreae Concha, Inula helenium, Biotae Semen, Schizandra chinensis, Cinnamomum loureirii, Cinnamomum loureirii, Disocorea batatas, Chinese matrimony vine, Mac The composition containing X extracted with one or more auxiliary herbs selected from snake's beard, GLYCYRRHIZAE RADIX, SCHIZANDRAE FRUCTUS or a solvent selected from water, lower alcohol, ethyl acetate, aromatic hydrocarbons and chlorinated hydrocarbons. Alzheimer's disease, vascular disease (multi-infarct dementia), a combination of Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, alcoholic dementia It has an excellent effect on the treatment of dementia.

Claims (4)

Dermis (Aurantii Nobilis Pericarpium), Hoelen, Ligusticum acutilobum, Uncariae Ramulus et Uncus, Acorus gramineus, Atractyodis Rhizoma, Angelica dahurica, Sanjoin (Zsemeny Spin) , Powdered Rehmanniae Radix Preparata, Cornus officinalis and raw paper (Polygala tenuifolia), or extracted with water, organic solvents or mixed solvents as main ingredients, and if necessary Cyperus rotundus, Pinellia ternata, Cnidium officinale, Peony lactiflora, Platycodi Radix, Ponciri Fructus, Keel, Ostreae Concha, Inula helenium 1 or more selected from Biotae Semen, Cinnamomum loureirii, Cinnamomum batatas, Chinese matrimony vine, Snake's beard, Licorice (GLYCYRRHIZAE RADIX), SchizandraRA FRUCTUS Or water thereof, Stroke, cerebral infarction, hypertension, Alzheimer's disease, vascular disease (multi-infarct dementia), Alzheimer's disease and multi-infarct dementia, containing X extracted with a solvent selected from alcohol, ethyl acetate, aromatic hydrocarbon, chlorinated hydrocarbon, A pharmaceutical composition that has excellent effects as a pharmaceutical composition for the treatment of Parkinson's disease, hypothyrodism and alcoholic dementia. According to claim 1, 50 to 500 parts by weight of dermis (Aurantii Nobilis Pericarpium), 50 to 500 parts by weight of Hoelen, 50 to 500 parts by weight of Ligusticum acutilobum, Uncariae Ramulus et Uncus 50 to 500 parts by weight Part, 50 to 500 parts by weight of Acorus gramineus, 50 to 500 parts by weight of Atractyodis Rhizoma, 50 to 500 parts by weight of Angelica dahurica, 50 to 500 parts by weight of Zizyphi Spinosi Semen, Rehmanniae Radix Preparata) 50 to 500 parts by weight, Cornus officinalis 50 to 500 parts by weight, raw paper (Polygala tenuifolia) 50 to 500 parts by weight or its extract, if necessary, here, Cyperus rotundus 50 to 500 parts by weight, 50 to 500 parts by weight of Pinellia ternata, 50 to 500 parts by weight of Cnidium officinale, 50 to 500 parts by weight of Paeonia lactiflora, 50 to 500 parts by weight of Platyodi Radix Ponciri Fructus) 50 to 500 parts by weight, 50-500 parts by weight of keel, 50-500 parts by weight of Ostreae Concha, 50-500 parts by weight of Inula helenium, 50-500 parts by weight of Biotae Semen, 50-500 parts by weight of Schizandra chinensis 500 parts by weight, 50 to 500 parts by weight of Cinnamomum loureirii, 50 to 500 parts by weight of Dosocorea batatas, 50 to 500 parts by weight of chinese matrimony vine, 50 to 500 parts by weight of snake's beard, licorice (GLYCYRRHIZAE RADIX) 50 to 500 parts by weight, SCHIZANDRAE FRUCTUS 50 to 500 parts by weight of one or more auxiliary drugs selected from 50 to 500 parts by weight or selected from water, lower alcohol, ethyl acetate, aromatic hydrocarbons, chlorinated hydrocarbons Stroke, cerebral infarction, hypertension, Alzheimer's disease, vascular disease (multi-infarct dementia), mixed Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, alcoholic dementia cure A pharmaceutical composition which has an excellent effect as a pharmaceutical composition. A paralytic formulated with a pharmaceutically acceptable excipient, adjuvant, diluent, isotonicity agent, preservative, ointment agent and solubilizing agent in the form of a pharmaceutically acceptable pharmaceutical formulation. It has excellent effects as a pharmaceutical composition for the treatment of cerebral infarction, hypertension, Alzheimer's disease, vascular disease (Multi-infarct dementia), Alzheimer's disease and multi-infarct dementia, Parkinson's disease, hypothyrodism, and alcoholic dementia Alzheimer's disease. Pharmaceutical preparations. According to claim 3, Cognex, Aricept, Memantine, Exelon, Dihydroebodiamine, Reminyl, Tacrine, Piracetam, Idebenone, Tocopherol (Vitamin E), It further contains one or more drugs selected from ginkgo biloba extract, estrogen, aspirin, ticlopidine, warfarin, seregline, and nisergoline, and includes pharmaceutically acceptable excipients, adjuvants, diluents, isotonic agents, preservatives, and suspending agents. , Stroke, cerebral infarction, hypertension, Alzheimer's disease, vascular disease (multi-infarct dementia), a combination of Alzheimer's disease and multi-infarct dementia, formulated in a pharmaceutically acceptable pharmaceutical formulation with a dissolution aid A pharmaceutical formulation having a pharmaceutical composition which has an excellent effect as a pharmaceutical composition for treating Alzheimer's disease, hypothyrodism and alcoholic dementia.