WO2005002568A1

WO2005002568A1 - The application of l-n-butylphthlide in preventing and treating dementia

Info

Publication number: WO2005002568A1
Application number: PCT/CN2004/000652
Authority: WO
Inventors: Yipu Feng; Xiaoliang Wang; Rongduan Wang; Ying Peng; Liying Yang; Jinghua Yang; Jingguo Sun; Zhankun Yang
Original assignee: Shijiazhuang Pharma. Group Zhongqi Pharmaceutical Technology (Shijiazhuang) Co., Ltd.; Institute Of Materia Medica, Chinese Academy Of Medical Sciences
Priority date: 2003-06-20
Filing date: 2004-06-18
Publication date: 2005-01-13
Also published as: CN1565441B; CN1565441A

Abstract

The invention disclosed the application of l-n-butylphthlide as formula (I) in preparing medicine for preventing and treating dementia, in special Alzheimer’s disease and vascular dementia.

Description

Application of L-n-butylphthalide in prevention and treatment of dementia Technical Field

The present invention relates to the use of L-n-butylphthalide and a composition containing L-n-butylphthalide to treat dementia.

Background technique

Alzheimer's disease (Alzheimer's disease, AD) is a progressive degenerative neuropathy characterized by clinical and pathological features. Its clinical manifestations are mainly memory loss (especially near memory), cognitive decline, mental retardation, and spatial disorientation. The pathological manifestations are β-amyloid peptide (Αβ) and other molecules, neurons and non-neuronal cells forming extracellular deposits to form senile plaques and the formation of intracellular neurofibrillary tangles (NFT). The prevalence of AD in China is between 0.2% and 5.98%, and the disease mostly occurs in people over 60 years of age. It increases with age, and it is estimated that there are more than 3.6 million AD patients in China. Investigation of the incidence of dementia in Beijing found that there were more vascular dementia (VD) than AD. (Zhang Mingyuan et al .: Incidence of dementia and Alzheimer's disease. Chinese Journal of Psychiatry 1998; 31 (4): 195-196) As China has entered an aging period, the number of dementia patients has increased year by year. In addition, the incidence of cerebrovascular disease is very high in the elderly. It is estimated that the incidence of post-stroke dementia is about 9-30.8%. Long-term cerebral insufficiency is also an important cause of vascular dementia. In short, patients with dementia are not only extremely distressed to themselves, but also burden the family and society. Therefore, it is extremely important to find effective drugs to delay and control the development of AD and VD. Alzheimer's disease (AD) is the leading cause of progressive cognitive decline in the elderly. The main pathological change is the formation of senile plaques and nerve fiber tangles with β-amyloid (A (3)) deposition as the core. Studies have shown that the cholinergic system in the brain is related to human learning and memory. AD The level of ACh in the patient's brain decreases, and the activity of choline acetyltransferase (ChAT), which catalyzes the synthesis of ACh, is low, and the degree of reduction is closely related to cognitive decline. In addition, the etiology of oxidative stress and inflammation involved in AD is also increasing It is valued. Αβ is composed of 39-43 amino acids and is a degradation product of β precursor protein (ΛPP). The range of Αβ deposition is closely related to nerve damage and cognitive impairment. Previous studies have confirmed: Continuous lateral ventricular perfusion ( icv) Αβ (1-40) or Αβ (1-42) can cause learning and memory impairment in rats (Ni 11 s. et al. β-Amyloid protein-induced Alzheimer's disease animal model. Neurosci. Lett. 1994; 170: 63-66), suggesting that memory impairment caused by A (3 aggregation in the brain can mimic the symptoms of AD patients. After research, L-positive Butylphthalide (L-NBP), can significantly improve mitochondrial function, improve cerebral microcirculation and energy metabolism, inhibit neuronal apoptosis, resist oxidative damage, inhibit inflammatory response, antithrombotic, reduce intracellular calcium, and inhibit glutamic acid The brain protective effect such as the release. For this purpose, continuous lateral ventricular perfusion (icv) A β (1-40) can be used as a model. The Morris water maze and biochemical methods are used to test the function of the test compound on the near memory and spatial memory function of animals and Effects on oxidative damage.

Vascular dementia (VD) is dementia due to cerebral dysfunction caused by cerebrovascular disease. Most of them are accompanied by multiple large cerebral arterial infarction or lacunar cerebral infarction or cerebral hypoperfusion. Decreased cerebral blood flow is related to the severity of dementia (Roman et al. Vascular dementia: diagnostic criteria for research studies. Neurology 1993; 43: 250-260) Chronic progressive cerebral insufficiency, which makes it impair oxygen and glucose and other necessary metabolism Substance utilization declines, resulting in oxidative damage, impaired mitochondrial function and nerve cell biosynthesis, impaired synaptic transmission, and finally leads to neuropathological changes, that is, neurodegeneration (Beal et al. Do defects in mitochondrial metabolism underlie the pathology of neurodegenerative disease Trends Neurosci. 1993; 16: 125-131) ₀ VD patients mainly present with progressive decline in near memory and spatial perception and cognitive impairment. The occurrence and development of vascular dementia and the cholinergic nervous system Information transmission is closely related to oxidative damage to nerve cells. Pathologically, the subcortical white matter is sparse in VD patients. A large number of studies have shown that ACh is considered to be an important neurotransmitter for learning and memory. Cholinergic pathway function in patients with AD is reduced As a decrease in neurotransmitter ACh levels This is one of the important reasons for its impaired memory and cognitive impairment (Toghi et al. Cerebrospinal fluid acetylcholine and choline in vascular dement ia of Binswanger and multiple small infarct types as compared with Alzheimer-type dementia. J. Neural Transm 1996; 103: 1211-1220). The tested compound has an enhancement effect on ChAT activity, indicating that it can increase the ACh level of cholinergic nerves, which is beneficial to improve memory function. In the past ten years, many laboratories have used Morris water Maze testing of near-memory and spatial memory in rats can sensitively reflect the damage to the central nervous system of the animal and its functional changes (Richard Morris. Developments of a water-maze procedure for studying spatial learning in the rat J. Neurosci Methods. 1984 11: 47-60), and observe the effect of drugs on the model use. Because dementia patients are mainly manifested as cognitive impairment, especially progressive impairment of near memory and spatial perception. Therefore, this model may be an ideal model to observe whether the drug has a therapeutic effect on Alzheimer's disease (AD) and VD. The continuous hypoperfusion in rats with bilateral common carotid artery occlusion (2-V0) model can simulate clinical vascular dementia caused by insufficient blood supply, so this method can reflect the role of drugs in treating dementia (Ni. JW et al. Neuronal damage and decrease of central acetylcholine level following permanent occlusion bi lateral common carotid arteries in rats.

Brain Res. 1995; 673: 290-296).

Apigenin (3-n-butylphthalide, Ag-1) reported by Coix seed in 1988 was a synthetic racemic 3-n-butylphthalide, which caused learning and memory impairment in rats It has an improving effect and has a protective effect on rat hippocampal cells (Yu Yanren et al .: The role of apigenin in enhancing learning and memory. Chinese Journal of Pharmacology 1988, 9 (5): 385-388). Since then, there have been reports that celery extract can improve the learning and memory of middle-aged and elderly mice. (Effects of Li Jing et al.'S celery extract on learning and memory of middle-aged and elderly mice. Chinese Traditional and Herbal Medicine 1996, 27 (2): 104-105; Liu Luosheng et al: An Study on the quality standards of Qingzhi Capsules, Journal of Shandong Medical University 2001, 39 (6): 562- 564), but so far no optical stereoisomer-L-n-butylphthalide (L-NBP) has been used to treat senile Report of dementia. Summary of the invention

In order to overcome the shortcomings of the prior art, the present invention provides the following formula (I)

Application of L-n-butylphthalide (hereinafter referred to as L-NBP) as an anti-dementia drug.

The L-n-butylphthalide used in the present invention is chemically synthesized to obtain racemic n-butylphthalide, and then chemically resolved into the L-isomeric n-butylphthalide. The spectrum is determined by nuclear magnetic resonance, mass spectrometry, and infrared. The analysis, especially using an Hp 5890 gas chromatograph and a chiral gas chromatography column (0 ^^ 1 (160--8)), proved that the optical and chemical purity of this product is a single optical stereoisomer-L-positive Butylphthalide (specific optical rotation> -66.49 °, optical purity> 98%, chemical purity> 98%), please refer to the Chinese patent for the resolution method Method for optically active 3-n-butylphthalide ", Application No. 99109673. 8, Publication No. CN1283621. The chemical structural formula of this product is the same as that of apigenin contained in edible celery and its seeds.

The present invention uses an internationally recognized method, that is, Morris water maze experiment to detect the near memory and spatial orientation of animals.

The present invention establishes a continuous hypoperfusion model by permanently blocking bilateral common carotid arteries (2-V0)

Mor r is s water maze method was used to detect the effects of L-NBP on the near memory and spatial function of animals. Biochemical methods were used to examine the effects of L-NBP on some indicators of oxidative damage and on the cholinergic nervous system. Given that behavioral changes caused by hypo-perfusion of the brain are accompanied by activation of glial cells, the white matter becomes thinner. Therefore, in this study, pathological and immunohistochemical methods were used, and glial cell fibrillary acidic protein (GFAP) and K-B staining (reflecting the pathological changes of neuromyelin sheaths) were used as indicators to observe the effects of drugs on them. The experiments of the present invention show that the left-n-butylphthalide of the present invention has a significant improvement effect on the near memory and spatial position dysfunction of rats with insufficient blood supply to the brain. The learning and retention experiments of the water maze test are often used to evaluate the spatial memory ability of hypoperfused rats. The experimental results show that in the water maze experiment, during the first day of training, there was no significant difference in latency between the groups, indicating that all animals were not familiar with the experimental operation on the following day. After 5 days of training, the search strategy of the sham operation group changed from edge and random to trend and straight, and the latency period (12.6 ± 3.3 seconds) was significantly shortened, indicating that the animals have certain memory and space after training. Directional force. The search strategy of the solvent control group did not change significantly, and remained marginal and random. The latency period (47.6 ± 5.88 seconds) was not significantly shortened, and there was a significant difference between the two groups (P <0.01).

The search strategy of the L-NBP10mg / kg group changed from edge and random to trend and linear, and the latency (26. 85 ± 5. 98 seconds) was significantly shortened, which was significantly different from that of the solvent control group (P <0.001 Two-factor analysis of variance), and there was no significant difference compared with the sham operation group, indicating that the dose group has a significant improvement on memory and spatial orientation. Other drugs such as DL-NBP1 0 mg / kg, DL-NBP30 mg / kg and D-NBP 30 mg / kg did not improve significantly. After 5 days of learning and training, platform exploration experiments were performed, and the island of safety was removed to test whether the rats had formed a spatial memory of the island of safety. With the exception of the solvent control group, the dwell time of all rats in the target quadrant was greater than 25%, indicating that a memory of the relative positions of the safety islands had been formed. The dwell time of the sham operation group was 17.73 ± 1.19 seconds, while the dwell time of the solvent group control group (14.40 ± 0.73 seconds) was significantly shortened. Statistical analysis by one-way analysis of variance> There were significant differences between the two groups (P <0.05). L-NBP1 0mg / kg group in Ping The residence time in the quadrant where the platform was located was significantly longer than that in the solvent control group (17.62 ± 1. 27 seconds), and there was a significant difference between the two groups (P <0.05). The DL-NBP group (10 mg / kg and 30 mg / kg) had no effect. In order to exclude the influence of the animal's athletic ability, it was determined that there was no difference in swimming speed between the groups. The above shows that only L-n-butylphthalide has a significant effect on the near memory and spatial position functional deficits in rats with insufficient blood supply to the brain, while meso and D-n-butylphthalide have no effect.

SOD is an important antioxidant enzyme. The activity of SOD in cortical tissue of normal control group is 100. 07 ± 3. 64 (NU / mg pro tein); the activity in hippocampal tissue is 57. 90 ± 7. 41 (U / mg pro te in). After permanent ligation of bilateral cervical movement in rats, hippocampal SOD activity was significantly increased compared to the control group (P <0.05), which may be a compensatory response. After treatment with L-NBP (10 mg / kg), the enzyme activity was significantly closer to normal levels (P <0.05). MDA is a marker of lipid peroxidation, which can reflect the degree of lipid peroxidation in the body and indirectly reflect the degree of cell damage. In this experiment, the content of MDA in the cortex of the model group increased by 19. 9%, which was significantly different from that of the normal control group (P <0.001). After treatment with L-NBP (10 mg / kg), the content of MDA in the cortex was significantly reduced by 20.7% (P <0.001). After permanent ligation of bilateral common carotid arteries in rats, the activity of ChAT in the cortex was significantly reduced, which was reduced by 34.4% compared with the normal control group (P <0.05), indicating that hypoperfusion can cause cholinergic nerve function Damaged. 1 L / NBP (10mg / kg) after 16 days of continuous administration, can increase the activity of ChAT in the cortical tissue compared with the model group by 37.1 »/. There is a significant difference (P <0. 05). From the results, the following conclusions can be drawn: The L-NBP 10mg / kg group can significantly improve memory impairment in the near memory and spatial position of 2-V0 animals, while racemic butylbenzene Phthalide and d-butylphthalide are not effective in improving memory dysfunction. The present invention is administered from the 10th day after 2-V0 (all the way to the 35th day). The purpose is to observe the effects of drugs on cerebral hypoperfusion Therapeutic effect of neuron degeneration in order to exclude the effect on acute hypoperfusion ischemia.

The present invention shows that L-NBP has a significant therapeutic effect on vascular dementia through pathological and immunohistochemical studies. After permanent ligation of bilateral common carotid arteries, neurons in the cortex and hippocampal CA1 area were significantly reduced, cell shrinkage and deep neuron staining were observed, and L-NBP (10mg / kg) significantly improved hypoperfusion induction after treatment. Neuron damage. It has been reported in the literature that bilateral carotid artery ligation can induce glial cell activation in the brain with thin white matter. White matter thinness is generally divided into 4 levels according to their severity: 0 level, normal; 1 level, nerve fiber staggered; 2 level, obvious vacuole formation; 3 level, myelinated fibers disappear. In our experiments, the visual bundle of the model group showed significantly thinner white matter than the normal control group, and a large number of vacuoles appeared. L-NBP (10 mg / kg) can significantly improve this situation after long-term administration, and the vacuole of the optic tract is significantly reduced. Immunohistochemical experiments found that GFAP-positive astrocytes were rarely detected in the hippocampus, caudate nucleus, and corpus callosum of the normal control group. However, after 4 weeks of bilateral common carotid ligation, many GFAP-positive Glia and microglia appeared. After treatment with L-NBP (10mg / kg), GFAP-positive glial cells were significantly reduced (see Figures 3 and 4). In short, L-NBP has an enhancement effect on ChAT activity, indicating that it can increase the ACh level of cholinergic nerves and is beneficial to improve memory function. In addition, L-NBP can significantly inhibit oxidative damage, indicating that L-NBP can reduce nerve cell damage. The pathological features of cerebral hypoperfusion caused by 2-V0 are sparse white matter, vacuoles, and femtocytosis (Narri. Et al. Chronic cerebral hypoperf us ion-induced neuropathological changes in rats. Jpn. J. Psychopharmacol. 1998; 18 : 181- 188), and L-NBP can improve these pathological changes. These mechanisms of action provide the basis for L-NBP to improve memory impairment caused by 2_V0 rats. Based on the above results, L-NBP has a significant therapeutic effect on vascular dementia. The experimental results of the present invention show that the levo-n-butylphthalide of the present invention has a significant improvement effect on the memory and spatial orientation disorder in rats caused by amyloid peptide (1-40). In the water maze experiment after continuous perfusion of A β in the rat ventricle, there was no significant difference in latency between the groups during the first day of training. After 5 days of training, the search strategy of the sham operation group changed from edge and random to trend and linear, and the latency (13.02 ± 2.77 seconds) was significantly shortened. However, the search strategy of the model group did not change significantly, and remained marginal and stochastic. The latency period (30.18 ± 4.81 seconds) was not significantly shortened, and the difference between the two groups was significant (P <0.01). After L-NPB treatment, the latency period of rats in the water maze test was significantly shortened. The search strategies for L-NBP 10mg / kg group and 30mg / kg group changed from marginal and random to trend and linear, and the latency was 27.28 ± 6.42 seconds and 25.88 ± 5.51 seconds, which were significantly different from the model group (P <0.05 two-factor analysis of variance), but not significantly different from the sham operation group, indicating that the rats treated with NBP were close to normal levels. See Figure 4. In the working memory test, the latency of the first experiment and the second to fifth experiments are shown in Figure 5A and B. Although in the first experiment, the latency of each group was not significantly different, but in the next 4 experiments, the latency of the sham operation group was 9.15 ± 0.91 seconds, and the latency of the model group was 14.05 ± 1.88 seconds. The latter has a significant latency. Prolonged, there is a significant difference between the two (P <0.01). L-NBP group can significantly improve working memory ability in a dose-dependent manner (P <0.01). ₀

L-NBP can also increase GSH-Px activity and decrease MDA content. GSH-Px is an important antioxidant enzyme. In our experiments, the activity of GSH-PX in the cortical tissue of the sham operation group was 15.86 ± 0.91 (U / mg pro te in); The activity in hippocampal tissue was 16. 19 ± 1. 19 (U / mg pro te in) „After continuous perfusion of rat A β (1-40), the activity of GSH_Px was compared with that of the sham operation group. The cortex and hippocampus were reduced by 29.5% and 42.4%, respectively, with significant differences (P <0.01 and P <0.001). After L-NBP treatment, the enzyme activity of the 30mg / kg group was significant. Elevated (P <0.01), L-NBP (10 mg / kg) also increased the activity of the enzyme, but there was no statistical significance. MDA is a marker of lipid peroxidation, which reflects the lipid peroxidation in vivo The degree of cell damage was indirectly reflected in this experiment. In this experiment, the levels of MDA in the cortex and hippocampus of rats after continuous perfusion of A β (1-40) increased by 25. 7% and 23. 6%, respectively. Compared with the surgery group, there were significant differences (P <0. 05 and P <0, 01). After L-NPB treatment, the MDA content in the cortex and hippocampus was significantly reduced, and the 10 mg / kg group was reduced by 28.4%. And 24.3% (P <0.05 and P <0.01), the effect of 30mg / kg is weaker than that of 10mg / kg group, but compared with the Αβ (1-40) model group, there are still significant differences (P <0. 05).

The experiment of the present invention can draw the following conclusions: Rats with continuous lateral ventricular perfusion Aβ (1-40) caused by memory disorder is a recognized model for observing the therapeutic effect of drugs on AD. From the above results, it can be seen that L-NPB not only has a significant effect on the model of vascular dementia caused by insufficient blood supply caused by 2-V0, but also has a near memory and memory impairment caused by continuous periventricular perfusion Aβ (1-40) in rats. There is also a significant improvement. This shows that L-NPB has a significant improvement effect on near memory and spatial memory disorders caused by different reasons. In addition, L-NBP has the effect of blocking oxidative damage (increasing GSH-Px activity and reducing MDA content), combined with its obvious brain protective effect, suggesting that L-NBP has the effect of treating senile dementia.

In summary, the present invention uses a Morr is water maze to detect the near-memory and spatial memory of rats, and the rat 2-V0 model is used for continuous hypoperfusion to simulate vascular dementia caused by insufficient blood supply in the clinic, so this method can fully reflect drug treatment The role of dementia. The experimental results of the present invention show that the L-NBP 10 mg / kg group can significantly improve the memory impairment in the near-memory and spatial positions of 2-V0 animals, while the racemic butylphthalide and d-butylphthalide both improve memory dysfunction. invalid. ACh is considered an important neurotransmitter for learning and memory. The cholinergic pathway function of AD patients is reduced, which is manifested by the decrease of the neurotransmitter ACh level, which is one of the important causes of impaired memory function and cognitive impairment. And L-NBP has an increase in the activity of ChAT, which can increase the ACh level of cholinergic nerves, which is beneficial to improve memory function. L-NBP can significantly inhibit oxidative damage, indicating that L-NBP can reduce nerve cell damage. The pathological features of cerebral hypoperfusion caused by 2-V0 are sparse white matter, increased vacuoles and glial cells, and L-NBP can improve these pathological changes. Therefore, the present invention is as The compound represented by the general formula (I) has a preventive and therapeutic effect on vascular dementia. Continuous lateral ventricular infusion of Aβ (1-40) caused learning and memory impairment in rats can mimic the symptoms of Alzheimer's patients. Morr is water maze was used to detect the memory function of near memory and spatial location in rats. L-NBP significantly improved near-memory and spatial-location memory disorders, while D-NBP and DL-NBP had no effect. GSH-Px is an important antioxidant enzyme; MDA is the degree of lipid peroxidation in the body and indirectly reflects the degree of brain cell damage. L-NBP can increase GSH-Px activity and reduce MDA content, indicating that L-NBP can block oxidative damage and protect brain damage, while D-NBP and DL-NBP have no effect. Therefore, the compound represented by the general formula (I) of the present invention, L-n-butylphthalide, has a preventive and therapeutic effect on Alzheimer's disease. In short, L-n-butylphthalide has the effect of preventing and treating senile dementia, while neither of racemic butylphthalide and d-butylphthalide has an effect.

The present invention therefore also relates to a pharmaceutical composition containing as active ingredient a compound of the invention and a conventional pharmaceutical excipient or adjuvant. 1-95 重量 ° /。 Usually the pharmaceutical composition of the present invention contains 0.1-95 weight ° /. Compounds of the invention.

Pharmaceutical compositions of the compounds of the invention can be prepared according to methods well known in the art. For this purpose, the compound of the present invention may be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants, if necessary, to prepare a suitable application form or dosage for use as human or veterinary medicine. form.

The compound of the present invention or a pharmaceutical composition containing the same can be administered in unit dosage form, and the administration route can be intestinal or parenteral, such as oral, intramuscular, subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum.

The compound of the present invention or a pharmaceutical composition containing the same can be administered by injection. Injections include intravenous, intramuscular, subcutaneous, intradermal, and acupoint injections.

The dosage form may be a liquid dosage form or a solid dosage form. For example, the liquid dosage form can be a true solution type, a colloid type, a particulate dosage form, an emulsion dosage form, or a suspension dosage form. Other dosage forms include tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, lyophilized powder injections and the like.

The compound of the present invention can be made into ordinary preparations, slow-release preparations, controlled-release preparations, targeted preparations and various microparticle delivery systems.

In order to form a unit dosage form into tablets, various carriers known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents such as starch, dextrin, sulfuric acid, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, white clay, microcrystalline cellulose, aluminum silicate Etc .; humectants and binders, such as water, glycerol, polyethylene glycol, ethanol, propanol, starch syrup, dextrin, syrup, honey, glucose solution, gum arabic, gelatin syrup, sodium carboxymethyl cellulose, Shellac, fluorenyl cellulose, phosphoric acid Potassium, polyvinylpyrrolidone, etc .; disintegrating agents, such as dry starch, alginate, agar powder, alginate, sodium bicarbonate and citric acid, carbonic acid 4, polyoxyethylene sorbitol fatty acid ester, dodecyl Sodium sulfonate, fluorenyl cellulose, ethyl cellulose, etc .; disintegration inhibitors, such as sucrose, glyceryl tristearate, cocoa butter, hydrogenated oil, etc .; absorption enhancers, such as quaternary ammonium salts, dodecyl Sodium sulfate, etc .; Lubricants, such as talc, silica, corn starch, stearates, boric acid, liquid paraffin, polyethylene glycol, etc. The tablets can also be further formed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer tablets and multilayer tablets.

For example, in order to make the administration unit into a pill, various carriers known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, kaolin, talc, etc .; binders such as gum arabic, tragacanth, gelatin, Ethanol, honey, liquid sugar, rice paste or batter, etc .; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecyl trace, cellulose, ethyl cellulose, etc.

For example, in order to make a capsule from an administration unit, the active ingredient of the compound of the present invention is mixed with various carriers as described above, and the resulting mixture is placed in a hard gelatin capsule or a soft capsule. The active ingredient of the compound of the present invention can also be made into a micro tincture, suspended in an aqueous medium to form a suspension, and can also be filled into a hard gelatin tincture or used as an injection.

For example, the compound of the present invention is prepared as an injection preparation, such as a solution, a suspension solution, an emulsion, a lyophilized powder injection, and the preparation may be aqueous or non-aqueous, and may contain one and / or multiple drugs Effectively acceptable carrier, diluent, binder, lubricant, preservative, surfactant or dispersant. For example, the diluent may be selected from water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxidized isostearyl alcohol, polyoxyethylene sorbitol fatty acid ester, and the like. In addition, in order to prepare an isotonic injection, an appropriate amount of sodium chloride, glucose, or glycerol may be added to the preparation for injection, and conventional co-solvents, buffers, pH adjusters, and the like may be added. These excipients are commonly used in the art

In addition, if necessary, a colorant, a preservative, a flavor, a flavoring agent, a sweetener, or other materials may be added to the pharmaceutical preparation.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or the pharmaceutical composition of the present invention can be administered by any known method of administration.

The dosage of the pharmaceutical composition of the compound of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, personality, and individual response of the patient or animal. The drug route, the number of administrations, and the purpose of treatment, so the therapeutic dose of the present invention can vary widely. Generally speaking, the dosage of the pharmaceutical ingredients in the present invention is well known to those skilled in the art. According to the actual drug amount contained in the final preparation in the compound composition of the present invention, appropriate adjustments can be made to meet the requirements of its therapeutically effective amount, and the prevention or treatment purpose of the present invention can be accomplished. The suitable daily dose range of the compound of the present invention is preferably from 0.1 to 100 mg / kg of body weight, and more preferably from 0.1 to 100 mg / day / person. The above dosages can be administered in a single dosage form or divided into several, for example, two, three, or four dosage forms, which are limited by the clinical experience of the administrating physician and dosing regimens including the use of other treatments.

The total dose required for each treatment can be divided into multiple or single doses. The compound or composition of the present invention may be taken alone, or used in combination with other therapeutic or symptomatic drugs and adjusted in dosage.

the term:

L-NBP: L-n-butylphthalide

2-V0 model: permanent bilateral carotid artery ligation

Figure 1. Water maze experiment, search path of rats on the fifth day of 1A; search path of rats in the platform exploration experiment on the fifth day of 1B. Figure 2. Effect of oral L-NBP after permanent bilateral carotid artery ligation on spatial memory impairment in rats in a water maze experiment. Figure 2A shows the change in latency during the training phase; Figure 2B shows the platform exploration experiment after the platform is removed. The rats swim freely for 60 seconds and stay in the quadrant (Q4) where the platform is located. All values are expressed as mean ± standard error. 12-14 rats in each group; #P <0.05 compared with sham group. * P <0.05 compared with the solvent control group (veh i c l e). Figure 3. H-E staining of the hippocampal CA1 area (A, B, C) and cortex (D, E, F) of the rat after hippocampal ligation of bilateral common carotid arteries at 5 weeks (40 times). Sham operation group (A, D); solvent control group (B,

E); L-NBP (10mg / kg) treatment group (C, F) Figure 4. · KB staining (40 times magnification of A, B, C) and tail of rat optic tract 5 weeks after bilateral common carotid artery ligation Microscopic image changes of nuclear GFAP immunohistochemical staining (D, E, F magnification 20x). Sham operation Group (A, D); Solvent control group (B, E); L-NBP (10mg / kg) treatment group (C, F) Figure 5. Continuous lateral ventricular perfusion of L-NBP in rats A β (1-40 Effects of memory impairment in water maze experiments. The figure shows the avoidance of latency changes during the training phase; all values are expressed as mean ± SD. 10 rats per group.

Figure 6. Effects of L-NPB on memory impairment caused by continuous lateral ventricular perfusion Aβ (1-40) in rats. Working memory tests (5 times a day) were performed on days 14-16 after Αβ (1-40) perfusion. Figure 5A shows the change in latency during the first experiment; Figure 5B shows the change in latency during the next 4 experiments. All values are expressed as mean ± standard error. 10 rats in each group. ## Ρ <0.01 compared with sham operation group; * P <0.05 compared with Αβ (1-4) model group. detailed description

1. Example 1 Treatment of senile dementia and vascular dementia by L-n-butylphthalide Materials and methods Reagents and medicines

L-, D-, DL-NBP are provided by the synthesis room of this firm. The optical and chemical purity is> 98%, and the optical rotation is -66.49, +66.88 and 0 degrees in order. Formulated with vegetable oil.

Instrument

Morris water maze automatic monitor, dark-proof box developed by the Institute of Materia Medica, Chinese Academy of Medical Sciences

Male Wistar rats, 10 weeks old, weighing about 280 grams, were placed in 1 cage per 5 animals, kept at room temperature at 23 ° C, and fed and drained freely. Rats were anesthetized with sodium pentobarbital (40mg / kg), bilateral common carotid arteries were exposed, and the capsule and vagus nerve of the common carotid arteries were carefully separated. In the hypoperfusion model group, bilateral common carotid arteries were ligated with 5-0 silk. The sham operation group underwent the same operation except that the common carotid artery was not ligated. Sprinkle a little sterilized crystal sulfa powder on the wound after surgery, and suture the skin. One month after the operation, a water maze experiment and a dark avoidance experiment were performed.

Experimental grouping and design

The rats were randomly divided into 8 groups of 10 rats each. 1) Sham operation group: Except that the common carotid artery is not ligated, The rest of the operations were the same as those of the hypoperfusion group; 2) the solvent control group: oral vegetable oil only; 3) the DL-NBP 10 mg / kg group; 4) the DL-NBP 30 mg / kg group; 5) the L-NBP 10 mg / kg Group; 6) L-NBP 30 rag / kg group; 7) D-NBP 30 mg / kg group; starting from the 10th day after surgery, drugs or solvents were administered. The water maze experiment was performed on the 29th-33th day after the operation, and the avoidance of the dark maggots was performed on the 34-35th day. Animals were sacrificed on day 36 for biochemical or pathological examination. In behavioral experiments, they were administered 40 minutes before the experiment. Oral administration of bilateral common carotid artery ligation in rats — avoid dark

10 20 29 30 31 32 33 34 35 36

Water maze t guillotine

Platform exploration

Water maze experiment

The Morr is water maze is mainly composed of a metal cylindrical pool (pool height 60cm, diameter 120cm) and automatic display, monitoring, recording device and safety island (platform 10cm in diameter). Fresh water was poured into the pool in advance, and then an aqueous solution of 1500 g of fresh milk powder was added to make the pool water opaque and milky, and the water level was higher than the platform 15. In this way, the animal cannot reach the platform through hearing, sight and smell in order to detect the animal's sensitivity to the spatial position. The water temperature is maintained at 23 ° C and 1 ° C. The pool is divided into 4 quadrants (east, south, west, and north), and the platform is placed in the center of the southwest quadrant. The swimming activity of each rat was monitored by a television and directly connected to a computer for processing and analysis. The water maze experiment was performed continuously for 5 days. Each rat received two trainings a day to find a platform, two times from the midpoint of the northeast quadrant and the northwest quadrant, with their heads facing the pool wall, and the interval between the two training sessions was 10 minutes. Record the time to find the platform (latency) and average the results of the two experiments. If the rat does not find the platform within 60 seconds, the incubation period is calculated as 60 seconds „Whether the platform is found within 60 seconds or not, the rat stays on the platform for 10 seconds. Put the rat on the platform before the first experiment begins Adaptation in 10 seconds. With the increase of the number of training, the latency of each group of rats to find a safe island shortens. After the last training, the exploration experiment is performed. The platform is removed and the rats swim freely for 60 seconds to find the platform. Record the cost of the rats in each quadrant of Time. The long time that the rats stayed in the quadrant where the original platform was located suggests that the rats already have memory for this spatial target.

We divide the search strategy into 4 categories: (1) edge type, rats move along the edge of the pool, no motivation to find; (2) random type, rats have no clear direction when searching; (3) trend type, rats have remembered The approximate location of the island of refuge was less than 4 turns before the island was found; (4) The straight line, the rat has clearly remembered the location of the island of safety and swam directly to the island of safety. The experimental results are expressed in terms of the time to find the safety island, that is, the latency and the search strategy. Detection of choline acetyltransferase, antioxidant enzymes, and MDA

The rats were decapitated and their brains were removed, and the cortex and hippocampus were peeled off on an ice bath. After weighing, they were quickly placed in liquid nitrogen and frozen for use. The sample was added with pre-cooled pH 7.0 potassium brick buffer solution 0.05mol- '(containing 0.5mol-L "' EDTA and 7% glycerol), and a 10% tissue homogenate was made in an ice bath. The protein quantification was based on the Lowry ratio. Colorimetric determination.

1. Determination of Choline Acetyl Transfer (IK ChAT): Add 0.5 mol-L " ^{1 of} sodium phosphate buffer, acetyl coenzyme AO.0062 mol-L" ¹ , 1.0 mol-L " ^{1 of} choline chloride into the reaction system , Sulfuric acid, neostigmine 76 μηιοΐ'ΐ ', NaCL3 mol'L— ¹ and EDTAO. Oil mol'L—', 0.5 mol'l / 'of chloric anhydride hydrochloride each 40μ1, and finally add distilled water to each tube 0.8ml. After incubating at 37 ° C for 5min, add 200μ1 tissue homogenate to each tube, then add each tube to boiling water for 2min, then add 2.5mmol'L- ¹ sodium arsenate 0.8ml, and centrifuge at 15000 X g for 3min at room temperature 2.0ml supernatant was added with 40μ1 3 mmol-L " ¹ 4- PDS, and incubated at 25 ° C for 15 minutes, and the OD value was determined = 324nm). ChAT activity was calculated and expressed as nmolCoA'SH / mg protein / hr.

2. The determination of superoxide dismutase (SOD) and malondialdehyde (MDA) was performed with reference to the kit instructions of Nanjing Jiancheng Reagent Company. Pathology and immunohistochemistry

Four to six animals were randomly selected from each group. After the behavioral experiment, pentobarbital anesthesia (100mg / kg ίρ) was anesthetized, and the skin and chest cavity were sequentially cut open to fully expose the heart. The left apex was cut open, and the perfusion needle was directed toward the main body. Insert in the arterial direction, clamp with hemostatic forceps, infuse normal saline, cut the right atrium in the lower part of the right atrial appendage, infuse normal saline for 15-20min (200-300ml), the effluent becomes clear, and change to «polyaldialdehyde PBS solution to continue After perfusion for 15-20min (150-200ml), the whole body of the animal is stiff and the liver becomes white, then the head is decapitated and the brain is removed, and the front part is cut with a razor In the telencephalon and the posterior cerebellum, 4% polyacetal was added for 48 h (paraffin section) or 20% sucrose polyfluorene solution was fixed for 48 h (frozen section).

Paraffin sections were fixed and embedded. After sectioning, hematoxylin-Eosin (HE) staining and K-B (Kluver-Barrera Luxol fast blue) staining were used for pathological examination. Immunohistochemical method was used to detect the change of GFAP content in brain tissue. Statistical Analysis

All results are expressed as mean ± standard error. Differences in latency between groups in the water maze experiment were compared using a two-factor analysis of variance with repeated measurements. Differences between groups were examined using / w ocLSD or Turkey ^^. The single-factor analysis of variance was used for the water maze platform exploration experiment. The dark avoidance experiment used Kruskal-Wallis test and Mann-Whitney U test. Biochemical measurements were performed using a one-way analysis of variance. P <0.05 was significant difference. Experimental results

Effects of L-NBP on learning and memory of water maze in rats In water maze experiments, learning and retention experiments are often used to evaluate the spatial memory ability of hypoperfused rats. During the first day of training, there were no significant differences between the groups. After 5 days of training, the search strategy of the sham operation group shifted from edge and random to trend and straight, and the latency (12.6 ± 3.34 seconds) was significantly shortened. The search strategy of the solvent control group did not change significantly, and remained marginal and random. The latency period (47.6 ± 5.88 seconds) was not significantly shortened, and the difference between the two groups was significant (P <0.01). The search strategy of the L-NBP10mg / kg group shifted from edge and random to trend and linear, with a significantly shorter latency (26.85 ± 5.98 seconds) compared with the solvent control group (P <0.001 two-way analysis of variance), There was no significant difference compared with the sham operation group. Other drugs such as DL-NBP 10 mg / kg, DL-NBP 30 mg / kg and D-NBP 30 mg / kg did not improve significantly, as shown in Figures Ⅰ, Β and Figure 2. After 5 days of learning and training, platform exploration experiments were performed, and the island of safety was removed to test whether the rats had formed a spatial memory of the island of safety. With the exception of the solvent control group, all rats had a dwell time in the target quadrant greater than 25%, indicating that memory of the relative positions of the safety islands had been formed. The dwell time of the sham operation group (17.73 ± 1.19 seconds) was significantly longer than that of the solvent control group (14.40 ± 0.73 seconds :). Statistics were analyzed by one-way analysis of variance. There are significant differences (P <0.05) in Figure 3. The residence time of L-NBP10mg / kg group in the quadrant where the platform was located was significantly longer than that of the solvent control group (17.62 ± 1.27 seconds, P <0.05). DL-NBP The groups (10 mg / kg and 30 mg / kg) had no effect. In order to exclude the influence of the animal's athletic ability, it was determined that there was no difference in swimming speed between the groups. The above shows that only L-n-butylphthalide has a significant effect on the near memory and spatial position functional impairment of rats with cerebral insufficiency, while racemic and d-n-butylphthalide have no effect. Effects on SOD, ChAT activity and MDA content SOD is an important antioxidant enzyme. The activity of SOD in the cortical tissue of the normal control group is 100. 07 ± 3. 64 (NU / mg pro tein); the activity in the hippocampal tissue is 57 90 ± 7. 41 (U / mg pro te in). After permanent ligation of bilateral common carotid arteries in rats, the activity of SOD in hippocampus was significantly increased compared with the control group (P <0.05), which may be a compensatory response. After treatment with L-NBP (10 mg / kg), the enzyme activity was close to normal levels (P <0.05). MDA is a marker of lipid peroxidation and can reflect the degree of lipid peroxidation in the body, indirectly. It reflects the degree of cell damage. In this experiment, the content of MDA in the cortex of the model group increased by 19.9%, which was significantly different from that of the normal control group (P <0.001). After treatment with L-NBP (10mg / kg), the content of MDA in the cortex was significantly reduced by 20.7% (P <0.001). After permanent ligation of bilateral common carotid arteries in rats, the activity of ChAT in the cortex was significantly reduced, which was reduced by 34.4% compared with the normal control group (P <0.05), indicating that hypoperfusion can cause cholinergic nerve function Damaged. After continuous administration of L-NBP (10 mg / kg) for 16 days, the activity of ChAT in the cortical tissue was increased by 37.1% compared with the model group, with a significant difference (P <0.05). (See Table 1). Table 1. Effects of L-NBP on SOD and ChAT activity and MDA content in cortex and hippocampus of hypoperfused rats (n = 7-9) SOD activity, MDA content, ChAT activity

(NU / rag protein) (nmol / mg protein) (% of sham operation) Brain cortex hippocampus cortex hippocampus cortex hippocampus sham operation group 100. 07 ± 3. 64 57. 90 ± 7. 41 3. 13 + 0. 10 3. 03 ± 0. 12 100. 0 + 13. 3 100. 0 ± 7. 2 Solvent control group 1 14. 42 ± 7. 82 81. 16 ± 6. 84 ° 3. 91 + 0. 22 '"3 63 + 0. 45 65. 6 ± 15. 1 '102. 6 + 15. 1

L-NBP 98. 84 ± 5. 53 57. 60 ± 3. 86 * 3. 10 + 0. 09 ™ 3. 42 ± 0. 12 104. 3 ± 8. 2 * 110. 0 ± 13. 5

(10rag / kg)

'P <0. 05, "* ρ <0. 001 compared with the sham operation group;-ρ <0. 05, ··· ρ <0. 001 compared with the solvent control group. For pathology and histochemical After permanent bilateral ligation of the common carotid artery, neurons in the cortex and hippocampal CA1 area were significantly reduced, cell shrinkage and deep neuron staining were observed in the model group, and L-NBP (10mg / kg) After treatment, the neuronal damage induced by hypoperfusion can be significantly improved. It has been reported in the literature that bilateral common carotid ligation can induce glial cell activation with thin white matter. White matter thinning is generally divided into 4 levels according to their severity: 0 level, normal; 1 level, nerve fiber staggered; 2 level, obvious vacuole formation; 3 level, myelinated fibers disappear. In our experiment, the normal control group of the model group showed obvious thin white matter, and a large number of vacuoles appeared. After long-term administration of L-NBP (10mg / kg), this situation can be significantly improved. Cavitation was significantly reduced. Immunohistochemical experiments found that GFAP-positive astrocytes were rarely detected in the hippocampus, caudate nucleus, and corpus callosum of the normal control group. However, after 4 weeks of bilateral common carotid ligation, many GFAP-positive Glia and microglia appeared. After treatment with L-NBP (10mg / kg), GFAP-positive glial cells were significantly reduced (see Figures 3, 4).

Experimental conclusions From the results, the following conclusions can be drawn: L-NBP 10mg / kg group can significantly improve memory impairment in near-memory and spatial locations of 2-V0 animals, while racemic butylphthalide and d-butylphthalide can improve memory Dysfunction is ineffective. In this study, administration was started 10 days after 2-V0 (all the way to day 35). The purpose was to observe the therapeutic effect of the drug on the neuronal degeneration caused by cerebral hypoperfusion, in order to exclude the acute hypoperfusion ischemia period. The impact of ₀

L-NBP can increase the activity of ChAT, indicating that it can increase the ACh level of cholinergic nerves, which is conducive to improving memory function. In addition, L-NBP can significantly inhibit oxidative damage, indicating that L-NBP can reduce nerve cell damage. The pathological features of cerebral hypoperfusion caused by 2-V0 are sparse white matter, vacuoles and glial cells (Narr i. Et al. Chron ic cerebra l hypoperf us ion-induced neuropa tho log ica l changes in ra t s. Jpn. J. Psychopharmaco l. 1998; 18: 181-188), and L-NBP can improve these pathological changes. These mechanisms of action provided the basis for L-NBP to improve memory impairment caused by ^2- V0 rats. Based on the above results, it is suggested that L-NBP may have obvious therapeutic and preventive effects on vascular dementia.

Example 2 Levo-n-butylphthalide can significantly improve the symptoms of senile dementia caused by β-amyloid peptide (1-40) Materials and Method

Reagents and medicines: L-NBP is provided by our synthesis laboratory and formulated with vegetable oil. (1-40) purchased from Sigma. Alzet cerebral micro-osmotic pump perfusion device was purchased from DURECT Company.

Apparatus: Morris water maze automatic monitor and experimental method, see above. Experimental model of learning and memory impairment in cerebral hypoperfusion rats: Male Wistar rats, 10 weeks old, weighing about 280 grams, 1 animal per cage, kept at room temperature at twenty three. C, eat freely and divert water. Rats were anesthetized with sodium pentobarbital (40 mg / kg), fixed on a stereotactic device in the abdominal position, the skin at the beginning was cut, and a cannula for perfusion Aβ (1-40) was implanted in the right ventricle. > According to the stereotactic atlas of the rat brain of Paxions and Watson, the implantation site was 0.3mm posterior to the posterior condyle, 1.1 recessed to the right, and 4.0 mm deep. The cannula is connected to a micro-osmotic pump. The micro-osmotic pump was placed on the back of the rat. Αβ (1-40) was dissolved in 35% acetonitrile / 0.1 trifluoroacetic acid, and continuous ventricular perfusion was performed at 300 pmol / day (i.c. V). The control group was perfused with only 35% acetonitrile / 0.1% trifluoroacetic acid. Previous experiments have shown that at this flow rate, the solvent does not cause behavioral and neurochemical changes in rats. Experimental grouping and design: Rats were randomly divided into 4 groups of 10 rats each. 1) Sham operation group: Rat brain ventricle is perfused with only 35% acetonitrile / 0.1 trifluoroacetic acid + solvent; 2) Model group: Αβ (1-40) + solvent; 3) Αβ (1-40) + L- NBP10mg / kg Group; 4) Αβ (1-40) + L-NBP 30 mg / kg group. Drugs and solvents were administered from day 2 after surgery. The water maze training test was performed on the 9th to 13th days after the operation, the platform exploration test was performed on the 13th day, and the working memory test was performed on the M-16th day. Animals were sacrificed on day 17, and brains were decapitated for biochemical determination. In behavioral experiments, they were administered 40 minutes before the experiment.

Αβ (1-40) continuous ventricular perfusion (300pmol / day) orally

2 ^ 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Water maze training test † * wood

Platform exploration test

Workbook 'Recall Quiz Water maze experiment: The water maze training test was performed on the 9th to 3rd days after i cv Α β (1-40), and the platform exploration test was performed on the 13th day. Working memory test (see above) is performed on days 14-16 after i cv Α β (1-40). The test is based on the animals having acquired a certain memory base after the training test and platform exploration test. The ability of animals to learn spatial memory quickly after the quadrant. The experimental process is similar to the standard water maze training experiment except that the platform changes position daily. Five experiments per day, the rats started swimming from five water points. The first experiment of each day is called the demonstration experiment. Rats are allowed to swim on the platform in the new position and stay for 10 seconds. The next four experiments, the platform position remains unchanged, but the quadrant of the water entry point is different. The latent period of working memory ability is the average of the 2nd to 5th experiments, and the working memory ability of each rat is calculated from the experimental average of 3 days. Biochemical experiment: The determination of malondialdehyde (MDA) and glutathione peroxidase (GSH-Pχ) was the same as the above experiment. Statistical analysis: All results are expressed as mean standard error. Differences in latency between groups in the water maze experiment were compared using a two-factor analysis of variance with repeated measurements. Differences between groups were tested using the / ws t Z? Oc LSD or Turkey test. Water labyrinth platform exploration test, working memory test, biochemical measurement using single factor analysis of variance. P <0. 05 is a significant difference. Experimental results

Effects of L-NBP on learning and memory of water maze in rats

During the first day of training, there were no significant differences between the groups. After 5 days of training, the search strategy of the sham operation group changed from edge and random to trend and straight, and the latency (1 3. 02 ± 2. 77 seconds) was significantly shortened. However, the search strategy of the model group did not change significantly, and remained marginal and stochastic. The latency (30. 18 ± 4. 81 seconds) was not significantly shortened, and the difference between the two groups was significant (P <0.01). After treatment with L-NBP, the latency of rats in the water maze test was significantly shortened. The search strategies for L-NBP1 0mg / kg group and 30mg / kg group changed from marginal and random to trending and linear, and the latency was respectively It is 27. 28 ± 6.42 seconds and 25. 88 ± 5. 51 seconds. Compared with the model group, there is a significant difference (P <0.05 two-way analysis of variance), but there is no significant difference compared with the sham operation group. L-NBP treated rats have approached normal levels, see Figure 4. In the working memory test, the latency of the first experiment and the second to fifth experiments are shown in Figure 5 Λ, B. Although there was no significant difference in latency between the groups in the first demonstration experiment, in the next 4 experiments, the artificial hand The latency of the operation group was 9.15 ± 0.91 seconds, and the latency of the model group was 14.05 ± 1.88 seconds. The latency period of the latter group was significantly longer, and there was a significant difference between the two (P <0.01). The L-NBP group can significantly improve

o

Memory ability (P <0.01).

L-NBP increases GSH-Px activity and decreases MDA content

GSH-Px is an important antioxidant enzyme. In our experiments, the activity of GSH-PX in the cortical tissue of the sham operation group was 15.86 ± 0.91 (U / mg protein); the activity in the hippocampal tissue was 16.19 + 1.19 (U / After continuous perfusion of Αβ (1-40) in mg protein λ rats, the activity of GSH-Px was reduced by 29.5% and 42.4% in the cortex and hippocampus, respectively, compared with the sham operation group, with significant differences (P <0.01 and P < 0.001). After treatment with L-NBP, the activity of the enzyme in the 30mg / kg group was significantly increased (P <0.01), and L-NBP (10rag / kg) also increased the activity of the enzyme, but there was no statistical significance. MDA is a marker of lipid peroxidation, which reflects the degree of lipid peroxidation in the body and indirectly reflects the degree of cell damage. In this experiment, the levels of MDA in the cortex and hippocampus after continuous perfusion of Aβ (1-40) in rats Increased by 25.7% and 23.6%, respectively, compared with the sham operation group (P <0.05 and P <0.01). After treatment with L-NBP, the levels of MDA in the cortex and hippocampus were significantly reduced, in the 10mg / kg group They were reduced by 28.4¾ and 24.3%, respectively (P <0.05 and P <0.01). The effect of 30mg / kg was weaker than that of 10mg / kg group, but compared with (1-40) Compared with the model group, there is still a significant difference (P <0.05), see Table 2. Table 1. L-NBP on cortical and hippocampal SOD activity in rats after continuous flow A β (1-40) And MDA content (n == 7)

Group GSH-PX MDA

(U / mg protein) (nmol / mg protein)

Cortex hippocampus cortex hippocampus sham operation group 15.8610.91 16.19 ± 1.19 1.10 ± 0.09 1.13 + 0.06

Model group11 · 18 ± 1.32 ^## 9.32 ± 1.01 ^### 1.48 ± 0.14 ^# 1.48 ± 0.12 ^##

L-NBP 11.73 ± 0.53 12.73 ± 1.34 1.0610.05 * 1.12 ± 0.07 "

(lOmgkg)

L-NBP 15.40 ± 1.03 ** 18.91 ± 1.16 ** 1.06 ± 0.10 * 1.15 ± 0.07 *

(30mg / kg)

'P <0.05, "P <0.01,"' P <0.001 compared with the sham operation group; 'P <0.05, "P <0.01 compared with the model group. Experimental discussion:

Memory impairment caused by continuous lateral ventricular perfusion of Aβ (1-40) in rats is a recognized model for observing the effects of drugs on AD treatment. From the above results, it can be seen that L-NBP not only has a significant effect on the model of vascular dementia caused by insufficient blood supply caused by 2-V0, but also on the near memory and spatial memory impairment caused by continuous lateral ventricular perfusion Aβ (1-40) in rats. There is also a significant improvement. This shows that L-NBP can significantly improve the near memory and spatial memory disorders caused by different reasons. In addition, L-NPB has the effect of blocking oxidative damage (increasing GSH-Px activity and reducing MDA content), combined with its obvious brain protective effect, suggesting that L-NBP has the effect of treating and preventing senile dementia.

Claims

Rights request

1. Application of L-n-butylphthalide as shown in the general formula (I) in the preparation of a medicine for preventing or treating dementia.

Q ₄ H

(I)

2. The use according to claim 1, wherein the dementia is Alzheimer's disease.

3. The application according to claim 1, wherein the dementia is vascular dementia.

4. Use according to claim 1, characterized in that the effective therapeutic dose of the compound is:

0.1 to 100 mg / kg / day.

5. A pharmaceutical composition, characterized in that it contains a therapeutically effective dose of L-n-butylphthalide as shown by the general formula (I) and a pharmaceutically acceptable carrier.

(I)

6. The pharmaceutical composition according to claim 5, wherein the pharmaceutical composition comprises a tablet, a capsule, a pill, an injection, a sustained-release preparation, a controlled-release preparation, and various microparticle delivery systems.