WO2016127831A1 - 绿原酸在制备治疗帕金森病的药物中的用途 - Google Patents

绿原酸在制备治疗帕金森病的药物中的用途 Download PDF

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WO2016127831A1
WO2016127831A1 PCT/CN2016/072765 CN2016072765W WO2016127831A1 WO 2016127831 A1 WO2016127831 A1 WO 2016127831A1 CN 2016072765 W CN2016072765 W CN 2016072765W WO 2016127831 A1 WO2016127831 A1 WO 2016127831A1
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chlorogenic acid
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drug
preparation
mice
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张洁
张梦甜
黄望
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四川九章生物科技有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate

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  • the invention belongs to the new medicinal use of chlorogenic acid, in particular to the use of chlorogenic acid in the preparation of a medicament for treating Parkinson's disease.
  • Parkinson's disease also known as tremor paralysis
  • PD Parkinson's disease
  • the main clinical pathological change is the selective death and loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc), resulting in insufficient striatum DA, leading to disorders of basal ganglia neuroregulatory function.
  • DA dopaminergic
  • SNc substantia nigra pars compacta
  • SNc substantia nigra pars compacta
  • mitigating oxidative stress damage can be used as a drug target to treat Parkinson's disease.
  • ketone bodies can improve the clinical symptoms of neurodegenerative diseases such as Parkinson's disease, show clear antioxidant protection, and have certain effects on neuroprotection.
  • Apoptosis in the substantia nigra pars compacta of patients with Parkinson's disease is an important pathological basis of the disease, inhibiting such apoptosis, and will also play a positive role in the treatment of
  • Chlorogenic acid CGA also known as caffeic acid, is a phenolic acid consisting of caffeic acid CA and quinic acid QA. Its chemical name is 3-o-caffeoyl quinine. Acid (3-o-caffeoylquinic acid CGA).
  • Chlorogenic acid is a phenylpropanoid synthesized by plants in the process of aerobic respiration through the pentose phosphate pathway intermediate. Its extraction (usually extracted from plants), synthetic technology mature, chlorogenic acid has been used in food, health care products, cosmetics and pharmaceuticals and other fields. Because it is widely found in a variety of common vegetables and fruits, it has a variety of biological activities, such as: cardiovascular protection, anti-oxidation, anti-UV and anti-radiation effects, anti-mutagenic and anti-cancer Use, antibacterial action, antiviral effect, lipid-lowering and hypoglycemic effect, immune regulation and so on. It has a wide range of applications in the fields of pharmaceutical, chemical and food.
  • the drug is a drug for alleviating oxidative stress damage.
  • the oxidative stress damage refers to the body's own pathological reasons, resulting in excessive production of free radicals, active oxygen or reactive nitrogen, and weakening of the body's antioxidant capacity, destroying the normal balance of oxidation-reduction of the body, resulting in imbalance The phenomenon.
  • the body now exhibits a reduction in enzymes in a range of antioxidant defense systems, such as antioxidant enzymes SOD, glutathione peroxidase (GSH-Px), and catalase (CAT).
  • SOD antioxidant enzymes
  • GSH-Px glutathione peroxidase
  • CAT catalase
  • the drug is a drug that up-regulates the HMGCS2 gene.
  • the HMGCS2 gene is a gene encoding trihydroxytrimethyl-CoA synthetase 2, and the expression amount thereof is positively correlated with the corresponding enzyme activity. It is also positively correlated with the amount of ketone body produced in the body.
  • the drug is a drug that activates trishydroxytrimethyl-CoA synthetase 2.
  • trishydroxytrimethyl-CoA synthetase 2 is a key enzyme for the formation of ketone bodies in the body, and it is widely present in various organs. Its activity is positively correlated with the level of ketone body formation.
  • the drug is a drug that promotes ketone body formation.
  • the ketone body is a product produced by incomplete oxidation of fatty acids.
  • the latest research results show that ketone bodies have energy supplementation and antioxidant protection effects in neurodegenerative diseases such as Parkinson's disease, and the mechanism is still unclear.
  • studies have confirmed that increasing the ketone body content in patients with such diseases can have a certain control effect on the condition.
  • the medicament is a preparation prepared by adding chlorogenic acid as an active ingredient and adding one or more pharmaceutically acceptable pharmaceutical excipients.
  • chlorogenic acid can be used as the only active ingredient or as one of the active ingredients.
  • the preparation is an oral preparation or an injection preparation.
  • the specific dosage form may be various pharmaceutical dosage forms such as powder, pill, oral solution, tablet, capsule, granule, lyophilized powder injection and the like.
  • the unit preparation of the preparation contains 1-10 mg of chlorogenic acid.
  • the clinical use dose of the preparation is: 1-100 mg/kg.
  • the test of the present invention shows that chlorogenic acid has a certain therapeutic effect on Parkinson's disease, and the results of the examples show that chlorogenic acid acts as a key enzyme for reducing oxidative stress damage and activating ketone body synthesis.
  • HMGCS2 capable of up-regulating the HMGCS2 gene, which is a gene encoding a downstream enzyme
  • the enzyme By up-regulating the HMGCS2 gene and activating its corresponding enzyme (trishydroxytrimethyl-CoA synthetase 2) activity, the enzyme synthesizes the key enzymes of the body's synthesis of ketone bodies, and its activity and content are positively correlated with the biosynthesis of ketone bodies.
  • chlorogenic acid can regulate the ketone body level of the model rats.
  • chlorogenic acid can inhibit the apoptosis of substantia nigra neurons in patients.
  • the pathological features of Parkinson's disease include oxidative stress damage and apoptosis of substantia nigra neurons.
  • the implementation of chlorogenic acid can reduce oxidative stress, inhibit the apoptosis of substantia nigra cells, and also chlorogenic acid. It can up-regulate the genes and enzyme activities corresponding to key enzymes of ketone body synthesis and regulate the formation of ketone bodies.
  • chlorogenic acid is beneficial to reduce the toxicity of substantia nigra and striatum neurons, activate the autophagy activity of the substantia nigra and striatum neurons, which is beneficial to the regulation of blood ketone content in the rat model of Parkinson's disease. It inhibits the apoptosis of substantia nigra cells in the rat model of Parkinson's disease, and has positive effects and therapeutic effects on the symptoms and pathology of Parkinson's disease.
  • Example 1 is a relative expression level of HMGCS2 gene in kidneys of each group of mice according to Example 2 of the present invention
  • Figure 2 is a relative value (100%) of trihydroxytrimethyl-CoA synthetase 2 activity of each group of mice in Example 2 of the present invention
  • Fig. 3 is a graph showing apoptosis flow detection data of mouse black blood cells of each group in Example 2 of the present invention.
  • Chlorogenic acid (Sichuan Jiuzhang Biotechnology Co., Ltd.) rotenone (SIGMA), dimethyl sulfoxide (DMSO, SIGMA), corn oil (Shandong Luhua Group Co., Ltd.), mouse ⁇ -SYN monoclonal antibody, SABC immune tissue Chemical kit purchase (Wuhan Dr. Germany), mouse tyrosine hydroxylase monoclonal antibody, rabbit microtubule-associated protein light chain 3 polyclonal antibody (Biyuntian), malondialdehyde content and superoxide dismutase, valley Glutathione peroxidase and catalase kit (Nanjing Jiancheng Bioengineering Co., Ltd.). The remaining reagents were domestically produced analytically pure.
  • NC group normal control group
  • PD model group rotenone
  • LYS group chlorogenic acid treatment group
  • rotenone was first dissolved in a very low volume of DMSO and then dissolved in corn oil (formed as a 2 mg/ml oil solution).
  • the rotenone group (PD model group) was injected subcutaneously with rotenone oil solution (3 mg/kg mouse body weight) in the neck and back, and the normal control group (NC group) was injected subcutaneously into the neck and back with an equal volume of corn oil (containing the corresponding volume of DMSO).
  • the chlorogenic acid treatment group (LYS group) was injected subcutaneously with rotenone in the neck and back, and chlorogenic acid was administered at a dose of 30 mg/kg/d. All mice were free to eat and drink during the experiment, temperature 22 ⁇ 2 ° C, relative humidity (60 ⁇ 15)%. Animals were observed daily for general status and recorded changes in animal behavior for 5 weeks.
  • WMT-200 Water Loss Video Analysis System uses computer vision related algorithms to track animal behavior in real time, and analyzes and records the time required for animals to search for targets after entering the water: placing the mice before the experiment.
  • the platform was adapted for 20s, then the mice were randomly placed into the pool from different quadrant walls. The mice were recorded on the platform for 5s and the recording time was 30s. (If the platform cannot be used within 30s, the mice will be guided to the platform for 10s. Finally, the mice are dried and placed in a rat cage. The mice were placed in the swimming pool, and the rats were divided into 1 group five times. The average escape time of the four groups of mice was measured to evaluate the spatial learning ability. The experiment was carried out as described above at the 2nd, 3rd, 4th and 5th week after the experimental treatment.
  • Oxidative stress means that when the body suffers from various harmful stimuli, the high activity molecules such as active oxygen radicals and reactive nitrogen radicals in the body are excessively produced, the degree of oxidation exceeds the removal of oxides, and the oxidation system and the antioxidant system are unbalanced, resulting in Tissue damage.
  • the body now exhibits a reduction in enzymes in a range of antioxidant defense systems, such as antioxidant enzymes SOD, glutathione peroxidase (GSH-Px), and catalase (CAT). More malondialdehyde (MDA) is produced.
  • SOD antioxidant enzymes
  • GSH-Px glutathione peroxidase
  • CAT catalase
  • MDA malondialdehyde
  • the brain was quickly decapitated by the neck-removal method, and the midbrain substantia nigra and striatum were quickly separated on the ice surface, and the glass homogenizer was made into a 10% tissue homogenate, and stored in a -80 ° C refrigerator.
  • the protein was quantified by BCA method in the experiment, and the activity of SOD, GSH-Px, CAT and MDA in brain tissue were detected by chemical colorimetry according to the kit instructions.
  • mice in the PD model group showed debilitating on the 10th day from the start of the experiment, the activity was slow, the hair was loose and dull, and the appetite was obviously reduced. On the 16th day, the body flexed, the movement decreased, and sometimes the tonic and tremor performance showed The typical behavioral characteristics of Jinsen indicate that the mice with Parkinson's disease have successfully modeled.
  • the other two groups of mice showed no body flexion, reduced exercise, and rigidity and tremor.
  • the daily body weight measurements of the mice in each group showed that the weight gain of the PD model group was slower than that of the other groups, but there was no significant difference between the groups.
  • mice The MDA, GSH-Px, SOD and CAT of the PD model group were significantly higher than those of the NC group (P ⁇ 0.05). There was significant difference between the data of the LYS treatment group and the PD model group (P ⁇ 0.05). There was no statistical difference between the NC group and the NC group (p>0.05). (3) striatum oxidative stress index in mice
  • the MDA, GSH-Px, SOD and CAT of the PD model group were significantly higher than those of the NC group (P ⁇ 0.05).
  • the data of the chlorogenic acid treatment group were significantly different from those of the PD model group (P ⁇ 0.05). There was no statistical difference between the two groups (p>0.05). (4) Analysis of neuropathic behavior of water labyrinth
  • chlorogenic acid can effectively improve the pathological symptoms of the rattan model of Parkinson's mouse, reduce the toxicity of substantia nigra and striatum neurons, and activate the autophagy activity of the substantia nigra and striatum neurons. It has a preventive and therapeutic effect on Parkinson's disease.
  • Example 2 Effect of chlorogenic acid on HMGCS2 gene and corresponding trihydroxytrimethyl-CoA synthetase 2 activity, ketone body content and substantia nigra cell apoptosis in Parkinson's model mice
  • Chlorogenic acid Metopa (Dobasuo tablets, Shanghai Roche Pharmaceutical Co., Ltd.), microplate reader (Finland Rabm mk3 microplate reader), real-time PCR instrument (American Applied Biosystems; ABI), total RNA extraction kit (Tiangen Biochemical Technology Co., Ltd.), TIANScript cDNA first strand synthesis kit (Tiangen Biochemical Technology Co., Ltd.), Annexin-FITC-PI double staining apoptosis detection kit (Biyuntian), mouse ketone body An ELISA kit (Sigma), a flow cytometer (Beckman), and the like were detected.
  • the PD model mice were randomly divided into 3 groups, named as: Treatment group (positive drug treatment group), chlorogenic acid treatment group (LYS group) and Parkinson's disease model control group (PD)
  • Treatment group positive drug treatment group
  • LYS group chlorogenic acid treatment group
  • PD Parkinson's disease model control group
  • the mice were observed and the body was found to have buckling, reduced exercise, and sometimes accompanied by tonic and tremor performance, indicating typical behavioral characteristics of Parkinson's disease, indicating that Parkinson's disease mice were made.
  • the model succeeded.
  • the chlorogenic acid treatment group and the Madopar treatment group were intragastrically administered at doses of 60 mg/kg and 125 mg/kg, respectively.
  • the PD model group and the normal control group were injected with an equal volume of physiological saline.
  • mice in each group were removed from the eyeballs, and the blood was placed in a test tube which was rinsed with EDTA, numbered separately, centrifuged at 2000 g/min for 15 min, and the supernatant was taken.
  • the blood ketone content of each group of mice was determined using a mouse ketone body detection ELISA kit.
  • mice were sacrificed by cervical dislocation, the kidneys of the mice were taken and divided into two parts, A and B, for the following experiments:
  • the experimental mouse kidney A tissue homogenate was taken, and 500 ⁇ l of the lysate to which ⁇ -mercaptoethanol was added was added thereto, and mixed for 5 minutes. 600 ⁇ l of RNase-free ddH 2 O and 10 ⁇ l of proteinase K were added to the homogenate, mixed and treated at 56 ° C for 20 min; mRNA was extracted strictly according to the specific steps of the total RNA extraction kit, and stored in a refrigerator at -80 ° C for use.
  • the first strand cDNA is synthesized by using the cDNA first strand synthesis kit and the mRNA solution extracted by the above steps.
  • the specific reverse transcription process was carried out according to the instructions, and the obtained first strand of cDNA was stored at -20 ° C for storage and set aside.
  • the combination of SYBER GREEN fluorescent dye and double-stranded DNA produces strong fluorescence. By detecting and recording the final fluorescence intensity, the total amount of DNA produced can be obtained.
  • the fluorescent dye, the cDNA product synthesized in (1) step (2), the HMGCS2 primer (upstream primer: GTATGGGCTTCTGTTCG, the downstream primer: GGCGTTGGTGGTATCTA) were uniformly mixed and then subjected to RT-PCR. The system was replaced with RNase-free water.
  • the cDNA products group was an NTC control. Each sample was set to a relative value of the ⁇ -actin primer set for relative quantification.
  • Trihydroxytrimethyl-CoA synthetase 2 is an enzyme corresponding to the HMGCS2 gene.
  • the activity of this method is tested in the following examples.
  • the specific experimental steps are as follows: Take the above-mentioned B mouse kidney tissue, add PBS homogenate and add anhydrous After acetonitrile was allowed to stand for 10 min, it was centrifuged at 5000 g/min for 20 min, and the supernatant was taken to use mouse trishydroxytrimethyl-CoA synthetase 2 ELISA. A kit for detecting the activity of an enzyme. Based on the measured values of the NC group, the ratio (%) of the detected values of the other groups of mice to the detected values of the NC group was used as the final data for statistical analysis.
  • Annexin-V-FITC-PI double staining apoptosis kit was used to detect the apoptosis of substantia nigra cells.
  • the specific experimental operation is as follows:
  • mice were sacrificed, the whole brain was removed by craniotomy, and then according to the substantia nigra of the mouse brain map, the substantia nigra tissue was quickly taken out and placed in Trizol lysate, and the removed substantia nigra tissue was well homogenized, followed by PBS. Gently resuspend the cells gently and count;
  • Annexin V-FITC was green fluorescence and PI was red fluorescence.
  • the blood ketone content of each group of mice in the examples showed that the blood ketone content of the mice in the PD model group was significantly lower than that in the NC group, and there was a significant difference between the two groups (p ⁇ 0.05). ). After treatment with chlorogenic acid, the blood ketone content was significantly improved, and the value was not statistically different from that of the NC group (p>0.05), while the blood of the positive control drug Madopar was increased. The height was not obvious enough, and there was still a significant difference (p ⁇ 0.05) from the normal group.
  • the specific experimental results are shown in Table 1.
  • the enzyme activity in the kidneys of each group of mice was detected by using an ELISA kit of trishydroxytrimethyl-CoA synthetase 2, and the remaining groups of mice were calculated based on the measured values of the NC group.
  • the specific experimental results are shown in Figure 2: the LYS group can significantly enhance the activity of trishydroxytrimethyl-CoA synthetase 2, and its value is significantly higher than that of other groups of mice. Difference (p ⁇ 0.05).
  • Apoptosis of mesencephalic nigral cells is one of the important indications of Parkinson's disease.
  • the results of apoptosis detection of mouse nigral cells are shown in Figure 3.
  • the apoptosis of mouse substantia nigra cells in PD group is more serious. Very obvious pathological symptoms of Parkinson's disease.
  • the percentage of apoptosis in mice treated with domeba and chlorogenic acid was significantly down-regulated. Both treatment groups showed positive anti-apoptotic ability.
  • the experimental results are shown in Figure 3:
  • the D4 region represents apoptotic cells, the D3 region represents normal cells, and the rest are damaged or dead cells.
  • This example examines the effect of chlorogenic acid on various indicators of the Parkinson's mouse model induced by rotenone.
  • the results showed that chlorogenic acid can significantly increase the expression level of HMGCS2 gene in the kidney of Parkinson's mice, and further showed a strong activation effect on HMGCS2 corresponding trihydroxytrimethyl-CoA synthetase 2, which is in vivo.
  • the key catalytic enzyme for ketone body synthesis, ketone body has protective and anti-oxidative effects on patients with Parkinson's disease.
  • the blood ketone test results of each group also show that chlorogenic acid can effectively regulate the blood ketone content of mice, prompting It returns to normal levels.
  • chlorogenic acid has a certain ability to resist the apoptosis of substantia nigra and has a certain control effect on the progression of Parkinson's disease.
  • Chlorogenic acid provides a theoretical basis for the research and development of drugs for the treatment of Parkinson's disease.
  • the chlorogenic acid raw material used in the present embodiment is obtained by extracting and purifying from honeysuckle, and the purity is 99.52%.
  • the above prescription was completely dissolved in water for injection, filtered, and then finely filtered with a 0.22 ⁇ m sterile microfiltration membrane to adjust the pH. After the conventional operation of the sterile powder injection, 1000 ml of 2 ml powder injections were prepared. Contains 40mg of chlorogenic acid.
  • the chlorogenic acid used in the present example was extracted and purified from Eucommia ulmoides leaves, and the purity was 98.33%.
  • the chlorogenic acid used in the present example was extracted and purified from the burdock leaves, and the purity was 99.04%.
  • each oral solution is 10mL, containing green
  • the original acid is 200mg.
  • the chlorogenic acid used in the present example was extracted and purified from honeysuckle, and the purity was 98.37%.
  • a chlorogenic acid tablet is prepared by a wet granule tableting method.
  • the chlorogenic acid used in the present example was obtained by extraction and purification from Eucommia ulmoides leaves, and the purity was 99.25%.
  • each capsule contains 50mg of chlorogenic acid.
  • the chlorogenic acid used in the present example was obtained by extraction and purification from Eucommia ulmoides leaves, and the purity was 98.74%.
  • Povidone K30 was taken and water for injection was added to prepare a solution. After the prescription amount of chlorogenic acid, mannitol and sucrose were uniformly mixed, the povidone K30 solution was added to prepare a soft material. According to the conventional preparation process of the granules, the granules are obtained after sieving, drying and granulating the soft materials. The granules were dispensed under aseptic conditions to prepare 400 bags of granules, each bag containing 500 mg of chlorogenic acid.
  • the chlorogenic acid raw material used in the present embodiment is obtained by extracting and purifying the burdock leaves, and the purity is 98.82%.

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Abstract

本发明提供绿原酸在制备治疗帕金森病的药物中的用途。

Description

绿原酸在制备治疗帕金森病的药物中的用途 技术领域
本发明属于绿原酸的新的药用用途，尤其涉及绿原酸在制备治疗帕金森的药物中的用途。
背景技术
帕金森病(PD)又名震颤麻痹，是最常见的神经退行性疾病之一。临床上的主要病理改变是中脑黑质致密部(SNc)多巴胺(DA)能神经元选择性的死亡、缺失，导致纹状体DA不足，从而导致基底节神经调节功能的紊乱，其发病的病因，截至目前尚未清楚。然而研究已经证实，该病与氧化应激和线粒体功能异常等具有密切关系，因此，减轻氧化应激损伤可以作为药物作用靶点来治疗帕金森病。此外，最新的研究显示，酮体可以改善帕金森病等神经系统退行性疾病的临床症状，显示出明确的抗氧化保护作用，并且在神经保护方面有一定的效果。帕金森病患者的黑质致密区细胞凋亡，是该病的重要病理基础，抑制此种凋亡，对该病的治疗也会起到积极的作用。
绿原酸(chlorogenic acid CGA)又名咖啡鞣酸，是由咖啡酸(caffeic acid CA)和奎尼酸(quinic acid QA)组成的缩酚酸，其化学名为3-o-咖啡酰奎尼酸(3-o-caffeoylquinic acid CGA)。
CAS No.：327-97-9
CAS Name：
[1S-(1a,3b,4a,5a)]-3-[[3-(3,4-Dihydroxyphenyl)-1-oxo-2-propenyl]oxy]-1,4,5-trihydroxycyclohexanecarboxylic acid
系统名：1，3，4，5-四羟基环己烷羧酸-(3，4-二羟基肉桂酸酯)
化学结构式：
Figure PCTCN2016072765-appb-000001
绿原酸是植物在进行有氧呼吸的过程中，经磷酸戊糖途径中间产物合成的一种苯丙素类物质。其提取(通常从植物中提取)、合成技术成熟，绿原酸已经被开放应用于食品，保健品，化妆品和药品等多个领域。由于它广泛的存在于常见的各种蔬菜水果中，具有多种生物活性，如:心血管保护作用、抗氧化作用、抗紫外及抗辐射作用、抗诱变及抗癌作 用、抗菌作用、抗病毒作用、降脂降糖作用、免疫调节作用等。在医药化工和食品等领域都具有广泛的应用。
发明内容
本发明的目的在于：提供绿原酸的新的药用用途，尤其是绿原酸在制备治疗帕金森的药物中的用途。
本发明目的通过下述技术方案来实现：
绿原酸在制备治疗帕金森病的药物中的用途。
其中，所述的药物是减轻氧化应激损伤的药物。
上述方案中，所述的氧化应激损伤是指机体由于自身的病理原因，导致自由基、活性氧或活性氮产生过多和机体抗氧化能力减弱，破坏机体氧化-还原的正常平衡，造成失衡的现象。机体此时表现出一系列抗氧化防御系统的酶减少，如抗氧化酶SOD、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)等的减少。而产生的自由基等又通过氧化反应，产生较多的丙二醛(MDA)，因而在氧化应激损伤时，机体的MDA值又会明显升高。
其中，所述的药物是上调HMGCS2基因的药物。
上述方案中，所述的HMGCS2基因是编码三羟基三甲基辅酶A合成酶2的基因，其表达量和对应的酶活性成正相关。与机体的酮体产生量也呈正相关。
其中，所述的药物是激活三羟基三甲基辅酶A合成酶2的药物。
上述方案中，三羟基三甲基辅酶A合成酶2是机体内酮体生成的关键酶，它广泛的存在于各种脏器中。其活性与机体酮体的生成水平呈正相关。
其中，所述的药物是促进酮体生成的药物。
上述方案中,酮体是脂肪酸不完全氧化所产生的产物，最新的研究结果显示，酮体对于神经退行性疾病，如帕金森病中具有能量补充和抗氧化保护作用，其中机制尚不清楚，然而研究已经证实，增加此类疾病患者体内的酮体含量，能够对病情产生一定的控制作用。
其中，所述的药物是由绿原酸为有效成分，加入一种或多种药学上可接受的药用赋形剂制备而成的制剂。
上述方案中，绿原酸既可以作为唯一有效成分，也可以作为有效成分之一。
其中，所述的制剂是口服制剂或注射制剂。
上述方案中，具体剂型可以为散剂、丸剂、口服溶液剂、片剂、胶囊剂、颗粒剂、冻干粉针剂等等各种药用剂型。
其中，所述的制剂的单位制剂含绿原酸1-1000mg。
其中，所述的制剂的临床使用剂量为：1-100mg/kg。
前述本发明主方案及其各进一步选择方案可以自由组合以形成多个方案，均为本发明可采用并要求保护的方案，本领域技术人员在了解本发明方案后根据现有技术和公知常识可明了有多种组合，均为本发明所要保护的技术方案，在此不做穷举。
本发明的有益效果：本发明试验表明，绿原酸对于帕金森病具有一定的治疗效果，实施例的结果显示，绿原酸通过减轻氧化应激损伤、激活酮体合成的关键酶三羟基三甲基辅酶A合成酶2活性、调节酮体生物合成和抑制黑质细胞凋亡等方面，对帕金森病的症状和具体的病理状态进行改善和治疗。
具体而言：
①绿原酸可以降低帕金森患者的氧化应激损伤程度；
②能够上调HMGCS2基因，该基因是编码合成下游酶的基因；
③通过上调HMGCS2基因，激活其所对应的酶(三羟基三甲基辅酶A合成酶2)活性，该酶使机体合成酮体的关键酶，其活性和含量与酮体的生物合成成正相关。
④增加机体酮体水平，对于帕金森病患者具有良好的疗效，绿原酸可以调节上调模型大鼠的酮体水平。
⑤绿原酸可以抑制患者黑质神经元细胞的凋亡。简而言之，帕金森发病的病理特征包括氧化应激损伤和黑质神经元细胞凋亡等，实施例证明绿原酸可以降低氧化应激，抑制黑质细胞凋亡，此外绿原酸还能上调酮体合成关键酶对应的基因和酶活性，调节酮体的生成。
综上，绿原酸有益于降低黑质及纹状体神经元的毒性，激活中脑黑质及纹状体神经元的自噬活性，有益于帕金森病鼠模型的血酮含量调节，有益于抑制帕金森病鼠模型脑黑质细胞的凋亡，对于帕金森病的病症和病理等具有积极的改善和治疗作用。
附图说明
图1是本发明实施例2的各组小鼠肾脏中HMGCS2基因的相对表达量；
图2是本发明实施例2的各组小鼠的三羟基三甲基辅酶A合成酶2活性的相对值(100％)；
图3是本发明实施例2的各组小鼠黑质细胞的凋亡流式检测数据图。
具体实施方式
下列非限制性实施例用于说明本发明。
实施例1.绿原酸对帕金森模型小鼠的体内药效学考察
实验材料
1.1实验试剂和仪器
绿原酸(四川九章生物科技有限公司)鱼藤酮(SIGMA)，二甲基亚砜(DMSO,SIGMA)，玉米油(山东鲁花集团有限公司)，鼠α-SYN单克隆抗体、SABC免疫组织化学试剂盒购(武汉博士德公司)，鼠酪氨酸羟化酶单克隆抗体、兔微管相关蛋白轻链3多克隆抗体(碧云天)，丙二醛含量及超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶试剂盒(南京建成生物工程有限公司)。其余试剂均为国产市售分析纯。
1.2动物造模及分组
本实验采用皮下注射鱼藤酮的方法构建帕金森病的小鼠模型。购买成年雄性C57BL/6小鼠30只，体重25-28g，由中科院上海实验动物中心提供。小鼠按体重随机平均分为正常对照组(NC组，n＝10)、鱼藤酮(PD模型组，n＝10)、绿原酸治疗组(LYS组，n＝10)。实验中，将鱼藤酮先溶于极低体积的DMSO，然后溶于玉米油(配成2mg/ml油溶液)。鱼藤酮组(PD模型组)每日于颈背部皮下注射鱼藤酮油溶液(3mg/kg小鼠体重)，正常对照组(NC组)于颈背部皮下注射等体积的玉米油(含相应体积的DMSO)，绿原酸治疗组(LYS组)每日于颈背部皮下注射鱼藤酮,同时按剂量30mg/kg/d灌胃绿原酸-。实验期间所有小鼠自由饮食和饮水，温度22±2℃，相对湿度(60±15)％。每日观察动物一般状态表现并记录动物行为变化，持续5周。
2.实验检测指标
2.1WMT-200水迷路
WMT-200水迷路视频分析系统(成都泰盟科技有限公司)采用计算机视觉相关算法，实时跟踪动物的行为状态，并分析和记录动物入水后搜索目标所需的时间：实验前将小鼠置于站台上适应20s，随后将小鼠随机从不同象限面壁置入池内，小鼠登上站台5s后终止记录，最长记录时间为30s(若30s内不能上台，引导小鼠登上站台适应10s，最后将小鼠擦干放人鼠笼)。如此将小鼠置入游泳池，5次为1组，分别测量四组小鼠平均逃逸时间，以评价其空间学习能力。在实验处理后第2、3、4和5周按上述步骤进行实验。
2.2氧化应激指标检测
氧化应激是指机体在遭受各种有害刺激时,体内高活性分子如活性氧自由基和活性氮自由基产生过多,氧化程度超出氧化物的清除,氧化系统和抗氧化系统失衡,从而导致组织损伤。机体此时表现出一系列抗氧化防御系统的酶减少，如抗氧化酶SOD、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)等的减少。而产生较多的丙二醛(MDA)。对SOD、GSH-Px、CAT和MDA进行检测，可以反应出机体氧化应激损伤的程度，对于帕金森病的病程发展等具有指导意义。
5周行为检测后，采用脱颈法迅速断头取脑，在冰面上迅速分离出中脑黑质和纹状体，玻璃匀浆器制成10％组织匀浆，-80℃冰箱保存。实验时用BCA法定量蛋白，按照试剂盒说明书，采用化学比色法检测脑组织中SOD、GSH-Px、CAT活性及MDA含量。
3.实验结果
3.1各组小鼠的行为状态及体重
PD模型组小鼠从实验开始第10天出现精神萎靡，活动缓慢，毛发松散而无光泽，食欲明显减退，第16天开始相继出现身体屈曲，运动减少，时而伴有强直、震颤表现，显示帕金森的典型行为特征，表明帕金森病小鼠造模成功。其余2组小鼠未出现身体屈曲，运动减少，强直、震颤表现。对各组小鼠每日体重测量发现,PD模型组小鼠体重增长较其它各组缓慢，但组间比较均无显著性差异。
3.2各组小鼠脑黑质氧化应激指标
对各组小鼠的黑质氧化应激指标的测定结果显示，PD模型组的小鼠，其被测定的各项生化指标均偏离NC组的小鼠所测定的值。LYS治疗组的小鼠各项氧化应激指标均得到了较大程度的恢复。所测定的结果如表1所示：
表1小鼠中脑黑质氧化应激指标(
Figure PCTCN2016072765-appb-000002
n＝10)
Figure PCTCN2016072765-appb-000003
与NC组比较*p＜0.05；与PD模型组比较△p＜0.05
PD模型组的MDA、GSH-Px、SOD、CAT均显著高于NC组(P<0.05)，LYS治疗组的各组数据与PD模型组之间均有显著性差异(P<0.05),而与NC组之间没有统计学差异(p>0.05)。(3)小鼠右侧纹状体氧化应激指标
小鼠右侧纹状体氧化应激指标的测定结果如表2所示：
表2小鼠右侧纹状体氧化应激指标(
Figure PCTCN2016072765-appb-000004
n＝10)
Figure PCTCN2016072765-appb-000005
与NC组比较*p＜0.05；与PD模型组比较△p＜0.05
PD模型组的MDA、GSH-Px、SOD、CAT均显著高于NC组(P<0.05)，绿原酸治疗组的各组数据与PD模型组相比均有显著性差异(P<0.05)，而与NC组之间没有统计学差异(p>0.05)。(4)水迷路神经行为分析
实验处理后第2、3、4和5周末，分别测量小鼠平均逃逸时间,结果显示，PD模型组小鼠在第3周末平均逃逸时间显著高于NC组(P<0.05)，第4和5周末差异极显著(P<0.01)。而LYS治疗组小鼠在第3周末平均逃逸时间显著低于PD模型组(P<0.05)，第4和5周末差异极显著(P<0.01)，实验具体结果如表3所示：
表3小鼠在实验处理后各周末平均逃逸时间(
Figure PCTCN2016072765-appb-000006
n＝5)
Figure PCTCN2016072765-appb-000007
与NC组比较*p＜0.05，**p＜0.01；与PD模型组比较△p＜0.05，△△p＜0.01
(5)TH、α-SYN和LC-3B免疫阳性反应
采集图像统计分析发现，PD模型组小鼠黑质TH阳性细胞数显著低于NC照和LYS组(P<0.01)，α-SYN,LC3-B阳性细胞数显著高于NC组和LYS组(P<0.01)。LYS治疗组与PD模型组比较，TH阳性神经元数显著增加，α-SYN，LC3-B阳性神经元数显著下降(P<0.05)，具体实验结果如表4所示：
表4小鼠中脑黑质致密区TH、α-SYN及LC3-B细胞数(
Figure PCTCN2016072765-appb-000008
n＝10)
Figure PCTCN2016072765-appb-000009
与NC组比较**p＜0.01；与PD模型组比较△p＜0.05
采集图像统计分析发现，PD模型组小鼠纹状体TH免疫阳性细胞数显著低于NC照和LYS治疗组(P<0.01)，α-SYN，LC3-B阳性细胞数显著高于NC和LYS组(P<0.01)。LYS治疗组和PD模型组比较，TH免疫阳性细胞数及α-SYN、LC3-B阳性神经元数显著高于PD模型组，有显著性差(P<0.05)，具体实验结果如表5所示：
表5小鼠中脑纹状体TH、α-SYN及LC3-B细胞数(
Figure PCTCN2016072765-appb-000010
n＝10)
Figure PCTCN2016072765-appb-000011
与NC组比较**p＜0.01；与PD模型组比较△p＜0.05，
4统计学处理
连续型变量形式的实验数据以
Figure PCTCN2016072765-appb-000012
表示，2组数据间的比较用两样本t检验。采用SPSS13.0(SolutionsStatistical Package for the Social Sciences/Statistical Product and Service Solutions)统计软件，P<0.05为有统计学意义。
5.试论
综上所述，绿原酸可有效改善鱼藤酮制造的帕金森小鼠模型的病理症状，降低黑质及纹状体神经元的毒性，激活中脑黑质及纹状体神经元的自噬活性，对帕金森病具有预防和治疗的作用。
实施例2.绿原酸对帕金森模型小鼠肾脏HMGCS2基因及对应三羟基三甲基辅酶A合成酶2活性、体内酮体含量和黑质细胞凋亡状况的影响考察
1.实验材料
1.1实验试剂和仪器
绿原酸、美多芭(多巴丝肼片，上海罗氏制药有限公司)、酶标仪(芬兰雷勃mk3酶标仪)、实时荧光定量PCR仪(美国应用生物系统公司；ABI)、总RNA提取试剂盒(天根生化科技有限公司)、TIANScriptcDNA第一链合成试剂盒(天根生化科技有限公司)、Annexin-FITC-PI双染凋亡检测试剂盒(碧云天)、小鼠酮体检测ELISA试剂盒(Sigma)、流式细胞仪(Beckman)等。
1.2动物模型和分组
BALB/C小鼠30只，随机取出10只作为正常对照组(NC组)，每天注射等量的对照液(二甲基亚砜：10％聚乙二醇＝1:1)。另外30只小鼠，每天按1mg/kg的给药剂量，腹腔注射鱼藤酮，连续给药28天建成PD模型小鼠，将PD模型小鼠随机平均分为3组，分别命名为：美多芭治疗组(阳性药物治疗组)、绿原酸治疗组(LYS组)和帕金森病模型对照组(PD 模型组)，在连续注射鱼藤酮后的第28天，观察小鼠，发现小鼠身体屈曲，运动减少，时而伴有强直、震颤表现，显示帕金森的典型行为特征，表明帕金森病小鼠造模成功。在第29天至第36天绿原酸治疗组和美多芭治疗组，分别按60mg/kg和125mg/kg的给药剂量，灌胃给药。PD模型组和正常对照组，则注射等体积的生理盐水。
2.实验测定指标
2.1各组小鼠的血酮含量检测
于实验的第36天对各组小鼠摘除眼球取血，将所取血置于被EDTA润洗过的试管内，分别编号，2000g/min离心15min，取上清液。用小鼠酮体检测ELISA试剂盒测定各组小鼠的血酮含量。
2.2小鼠肾脏中HMGCS2基因和对应酶活性的检测
颈部脱臼法处死小鼠之后，取小鼠肾脏，分为A和B两份分别做以下实验：
(1)小鼠肾脏中HMGCS2基因的表达情况检测
①小鼠肾脏细胞总mRNA的提取
取实验小鼠肾脏A份组织匀浆，向其中加入500μl的已加有β-巯基乙醇的裂解液，混匀5min。向匀浆液中加入600μl的RNase-free ddH2O和10μl的蛋白酶K,混匀后56℃处理20min；严格按总RNA提取试剂盒的具体步骤提取mRNA，冻存于-80℃冰箱内备用。
②cDNA第一链的合成
本实施例采用cDNA第一链合成试剂盒和上述步骤提取得到的mRNA溶液合成对应的第一链cDNA。具体逆转录过程按照说明书操作，得到的cDNA第一链，冻存于-20℃进行保存、备用。
③RT-PCR定量
SYBER GREEN荧光染料与双链的DNA结合会产生很强的荧光，通过检测和记录最终的荧光强度，可以得到生成DNA的总量。在试管中加入荧光染料、(1)(2)步中所合成的cDNA产物、HMGCS2引物(上游引物：GTATGGGCTTCTGTTCG；下游引物：GGCGTTGGTGGTATCTA)混合均匀后进行RT-PCR检测本体系使用RNase-free water代替cDNA products组为NTC对照。每个样品均设β-actin引物组为相对值，进行相对定量。
(2)三羟基三甲基辅酶A合成酶2活性的检测
三羟基三甲基辅酶A合成酶2是HMGCS2基因所对应的酶，本实施例对其活性进行检测，具体实验步骤如下：取上述B份的小鼠肾脏组织，加入PBS匀浆后加入无水乙腈静置10min后，以5000g/min的转速离心20min，取上清液，使用小鼠三羟基三甲基辅酶A合成酶2ELISA 试剂盒，检测酶的活性。以NC组的检测值作为基准，其余各组小鼠检测值与NC组的检测值之比(％)作为最终数据进行统计和分析。
2.3小鼠黑质神经元的凋亡检测实验
本实施例采用Annexin-V-FITC-PI双染凋亡试剂盒来对黑质细胞的凋亡情况进行检测。具体实验操作如下：
①将小鼠处死后，立即开颅取出全脑，再根据小鼠脑图谱黑质定位，快速取出黑质组织并放入Trizol裂解液，将取下的黑质组织充分匀浆之后，用PBS轻轻充分重悬细胞并计数；
②取1×105个重悬的细胞，1000g离心5分钟，弃上清后，加入195μl Annexin V-FITC结合液轻轻与细胞混合；随后加入5μl Annexin V-FITC，轻轻混匀；再室温避光孵育10min；最后用1000g离心5分钟，弃上清，加入190μl Annexin V-FITC结合液轻轻重悬细胞；
③加入10μl碘化丙啶染色液，轻轻混匀，冰浴避光放置；分别设置FITC单染组和PI单染组，用于补偿的调节，此两组分别仅单染一种荧光染料；
④将处理好的细胞随即进行流式细胞仪检测，Annexin V-FITC为绿色荧光，PI为红色荧光。
3.实验结果
3.1各组小鼠的血酮含量检测结果
实施例中各组小鼠的血酮含量检测结果显示，PD模型组的小鼠，其血酮含量与NC组相比，有了明显的下降，两组之间具有显著性差异(p<0.05)。使用绿原酸治疗之后，其血酮含量有所显著改善，其值与NC组相比没有统计学差异(p>0.05)，而阳性对照药物美多芭治疗的小鼠，其血酮含量升高不够明显，与正常组之间仍然有显著性差异(p<0.05)，具体实验结果如表1所示。
表1不同组别小鼠血酮浓度的测定结果(mmol/L,
Figure PCTCN2016072765-appb-000013
)
Figure PCTCN2016072765-appb-000014
Figure PCTCN2016072765-appb-000015
与NC组相比*：p<0.05与PD模型组相比#：p<0.05
与美多芭组相比，×：p<0.05
3.2小鼠肾脏中HMGCS2基因和对应酶(三羟基三甲基辅酶A合成酶2)活性的检测结果
(1)各组小鼠肾脏中HMGCS2基因的相对定量检测结果
本实施例中，以β-actin作为内参基因，以HMGCS2基因相对于β-actin的表达情况作为最终统计的数值，得到的结果显示：PD模型组的小鼠肾脏组织中的HMGCS2基因表达水平，与NC组小鼠肾脏中的HMGCS2表达水平相比，有了明显的下调，说明所造帕金森小鼠模型中HMGCS2基因表达下调是其病理特征的一部分。而在用绿原酸治疗后的小鼠肾脏中，其HMGCS2基因的表达水平有了显著的上调，与模型组之间有显著性差异(p<0.05)，阳性对照药美多芭治疗组的小鼠肾脏组织中的HMGCS2基因水平也有少量的增高，与绿原酸治疗组的HMGCS2基因表达水平尚有显著性差异(p<0.05).具体实验结果如图1所示：
(3)各组小鼠肾脏中三羟基三甲基辅酶A合成酶2的相对活性检测
本实施例采用三羟基三甲基辅酶A合成酶2的ELISA试剂盒对各组小鼠肾脏中的该酶活性进行检测，并以NC组的测定值为基准，分别计算了其余各组小鼠相对于NC组的酶活性，具体的实验结果如图2所示：LYS组能够显著的增强三羟基三甲基辅酶A合成酶2活性，其值与其它组小鼠相比，均有显著性差异(p<0.05)。
3.3小鼠黑质细胞的凋亡检测实验结果
中脑黑质细胞的凋亡是帕金森病的重要指征之一，小鼠黑质细胞的凋亡检测实验结果如图3所示：PD组的小鼠脑黑质细胞凋亡较为严重，体现出非常明显的帕金森病理症状。多美芭和绿原酸治疗后的小鼠，其凋亡的百分比有了较大的下调，两个治疗组均显示出很积极的抗凋亡能力，实验结果如图3所示：其中，D4区域代表凋亡细胞，D3区域代表正常细胞，其余为破损或死亡的细胞
4统计学处理
连续型变量形式的实验数据以
Figure PCTCN2016072765-appb-000016
表示，2组数据间的比较用两样本t检验。采用SPSS13.0统计软件，P<0.05为有统计学意义。
5.结论
本实施例考察了绿原酸对鱼藤酮所致的帕金森小鼠模型的各项指标的影响情况。结果显示，绿原酸能显著的提高帕金森小鼠肾脏中HMGCS2基因的表达水平，进而对HMGCS2对应的三羟基三甲基辅酶A合成酶2也显示出较强的激活作用，该酶是体内酮体合成的关键催化酶，酮体对于帕金森病患者具有保护和抗氧化作用，对各组小鼠的血酮检测结果也显示，绿原酸能有效的调节小鼠的血酮含量，促使其恢复至正常水平。此外，对小鼠黑质细胞的凋亡检测实验发现，绿原酸具有一定的抵抗黑质细胞凋亡的能力，对于帕金森病的进程具有一定的控制作用，综上，所述实施例对于绿原酸作为治疗帕金森病的药物研究和开发，提供了理论基础。
实施例3：用绿原酸制备冻干粉针剂
1.绿原酸的提取：
本实施例中所用的绿原酸原料药，系由金银花中提取、纯化得到，纯度为99.52％。
2.绿原酸冻干粉针剂的制备
2.1处方：
Figure PCTCN2016072765-appb-000017
将以上处方完全溶解于注射用水中，过滤后，再用0.22μm的除菌微孔滤膜精滤，调节pH后，按照无菌粉针剂的常规操作共制成2ml粉针剂1000支，每支含绿原酸40mg。
实施例4：用绿原酸制备丸剂
1.绿原酸的提取
本实施例中所使用的绿原酸，系由杜仲叶中提取、纯化得到的，纯度为98.33％。
2.绿原酸丸剂的制备
2.1处方
Figure PCTCN2016072765-appb-000018
Figure PCTCN2016072765-appb-000019
2.2.制法：
取适量的聚维酮K30，用无水乙醇配制成溶液，再取处方量的绿原酸和淀粉，采用等量稀释法混合均匀之后，加入聚维酮K30的乙醇溶液中，充分搅拌后制得软材，采用搓丸法制得绿原酸丸剂1000粒，每粒丸剂含绿原酸1mg。
实施例5：用绿原酸制备口服溶液剂
1.绿原酸的提取
本实施例中所使用的绿原酸，系由牛蒡叶中提取、纯化得到，纯度为99.04％。
1.绿原酸口服溶液剂的制备
2.1处方
Figure PCTCN2016072765-appb-000020
2.2制法
取处方量的绿原酸和亚硫酸氢钠，溶解于10L注射用水中，按照口服液的常规制备工艺，过滤后，无菌灌装成1000支口服液，每支口服液为10mL，含绿原酸200mg。
实施例6：用绿原酸制备片剂
1.绿原酸的提取：
本实施例中所使用的绿原酸，系由金银花中提取、纯化得到，纯度为98.37％。
2.绿原酸片剂的制备
2.1处方：
Figure PCTCN2016072765-appb-000021
2.2制法：
本实施例采用制湿颗粒压片法制备绿原酸片剂。(1)按处方量取羟丙甲纤维素制成水溶液；(2)取处方量的绿原酸、淀粉和乳糖混合均匀后，加入羟丙甲纤维素水溶液，充分 搅拌均匀后制成软材；(3)将制备好的软材按常规的湿法制粒的操作规程，过筛、干燥和整粒后得到大小均一的颗粒；(4)将制得的颗粒与硬脂酸镁混合均匀后压片，共制成1000片剂，每片含绿原酸100mg。
实施例7：用绿原酸制备胶囊剂
1.绿原酸的提取：
本实施例中所使用的绿原酸，系由杜仲叶中提取、纯化得到的，纯度为99.25％。
2.绿原酸胶囊剂的制备：
2.1处方：
Figure PCTCN2016072765-appb-000022
2.2制法：
取处方量的绿原酸和糖粉，混合均匀，加入80％乙醇溶液制成软材，干燥，整粒后按照胶囊剂的常规制备工艺制备2000粒胶囊，每粒胶囊含绿原酸50mg。
实施例8：用绿原酸制备颗粒剂
1.绿原酸的提取
本实施例中所使用的绿原酸，系由杜仲叶中提取、纯化得到的，纯度为98.74％。
2.绿原酸颗粒剂的制备
2.1处方：
Figure PCTCN2016072765-appb-000023
2.2制法：
取聚维酮K30，加入注射用水，制成溶液。取处方量的绿原酸、甘露醇和蔗糖混合均匀之后，加入聚维酮K30溶液，制成软材。按照颗粒剂的常规制备工艺，对软材进行过筛、干燥和整粒之后，得到颗粒剂。在无菌条件下分装颗粒剂，制备400袋颗粒剂，每袋颗粒剂含绿原酸500mg。
实施例9：用绿原酸制备散剂
2.绿原酸的提取：
本实施例所用的绿原酸原料药，系由牛蒡叶中提取、纯化得到的，纯度为98.82％。
2.绿原酸散剂的制备：
2.1处方
纯度为98.82％的绿原酸1000g
2.2制法
取处方量绿原酸过筛后，按照散剂的常规制备工艺，无菌分装成含1000瓶/袋散剂，每瓶/袋散剂含绿原酸1000mg。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (10)

  1. 绿原酸在制备治疗帕金森病的药物中的用途。
  2. 如权利要求1所述的用途，其特征在于：所述的药物是减轻氧化应激损伤的药物。
  3. 如权利要求1所述的用途，其特征在于：所述的药物是上调HMGCS2基因的药物。
  4. 如权利要求1所述的用途，其特征在于：所述的药物是激活三羟基三甲基辅酶A合成酶2的药物。
  5. 如权利要求1所述的用途，其特征在于：所述的药物是促进酮体生成的药物。
  6. 如权利要求1所述的用途，其特征在于，所述的药物是抑制帕金森病患者脑黑质细胞凋亡的药物。
  7. 如权利要求1至6中任一权利要求中所述的用途，其特征在于：所述的药物是由绿原酸为有效成分，加入一种或多种药学上可接受的药用赋形剂制备而成的制剂。
  8. 如权利要求7所述的用途，其特征在于：所述的制剂是口服制剂或注射制剂。
  9. 如权利要求7或8所述的用途，其特征在于：所述的制剂的单位制剂含绿原酸1-1000mg。
  10. 如权利要求7、8或9所述的用途，其特征在于：所述的制剂的临床使用剂量为：1-100mg/kg。
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