WO2008037182A1

WO2008037182A1 - Use of ginsenoside rg1 for treatment of cerebral ischemia and/or diseases or symptoms induced by cerebral ischemia

Info

Publication number: WO2008037182A1
Application number: PCT/CN2007/002836
Authority: WO
Inventors: Juntian Zhang; Zhiwei Qu; Yong Cheng
Original assignee: Institute Of Materia Medica, Chinese Academy Of Medical Sciences
Priority date: 2006-09-28
Filing date: 2007-09-28
Publication date: 2008-04-03
Also published as: CN101152194A

Abstract

Use of ginsenoside Rg1 for prevention, alleviation and/or treatment of cerebral ischemia and/or diseases or symptoms induced by cerebral ischemia. Use of ginsenoside Rg1 in preparation of pharmaceutical compositions for prevention, alleviation and/or treatment of cerebral ischemia and/or diseases or symptoms induced by cerebral ischemia. Method of prevention, alleviation and/or treatment of cerebral ischemia and/or diseases or symptoms induced by cerebral ischemia, comprising administration a treatment effective amount of ginsenoside Rg1 containing pharmaceutical compositions to the subjects in need thereof.

Description

Use of ginsenoside Rg1 for diseases or symptoms caused by cerebral ischemia and/or cerebral ischemia

The present invention relates to ginsenoside Rgl for preventing, alleviating and/or treating diseases or symptoms caused by cerebral ischemia and/or cerebral ischemia, in particular, reducing infarct size, reducing brain edema, improving neurological symptoms and improving cognitive dysfunction. Use; the ginsenoside Rgl is also used for preparing a disease or symptom caused by preventing, alleviating and/or treating cerebral ischemia and/or cerebral ischemia, in particular, reducing infarct size, reducing brain edema, improving neurological symptoms and improving recognition Use of a pharmaceutical composition known to be dysfunctional. The present invention also relates to a method for preventing, alleviating and/or treating diseases or symptoms caused by cerebral ischemia and/or cerebral ischemia, in particular, reducing infarct size, reducing brain edema, improving neurological symptoms, and improving cognitive dysfunction, This includes administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising ginsenoside Rgl. Background technique

Ginsenoside Rgl is a symbolic component of ginseng and the most important active ingredient of ginseng. This ingredient can be extracted from roots, stems, leaves and American ginseng of ginseng and Tianqi. The existing process has reached the level of kilogram production. The synthesis was successful. Its structure and related biological activities have been widely reported in relevant textbooks or published literature.

It has a wide range of biological activities, and the inventors' previous work proves that Rgl is the main active ingredient of ginseng promoting wisdom. Rgl has an anti-aging effect on anti-aging, immune enhancement in older mice, and has anti-apoptotic effects on several models of apoptosis. Its mechanism of memory improvement is mainly: to enhance the activity of choline acyltransferase, thereby improving the synthesis of acetylcholine; improving synaptic efficiency and structural plasticity, increasing the expression of neurotrophic factor BDNF and the expression of Jif/268 gene, especially It can stimulate the reproduction and transformation of adult rat brain neural stem cells into Neurons. Provided experimental evidence for the treatment of Alzheimer's disease.

However, the use of ginsenoside Rgl for preventing and/or treating cerebral ischemia and improving the direction of memory disorders caused by cerebral ischemia has not been reported so far. Summary of the invention

One aspect of the invention relates to the use of ginsenoside Rgl in the manufacture of a medicament for the prevention, mitigation and/or treatment of a disease or condition caused by cerebral ischemia and/or cerebral ischemia.

In the present invention, the disease or symptom caused by preventing, alleviating and/or treating cerebral ischemia and/or cerebral ischemia is selected from the group consisting of reducing the area of cerebral infarction, reducing cerebral edema, improving neurological symptoms and improving cognitive dysfunction, and memory. obstacle.

In the present invention, the prevention refers to the suppression of the occurrence of a condition such as cerebral infarction or cerebral edema before the onset of cerebral ischemia, particularly before the onset of stroke. The remission refers to reducing the area of cerebral infarction, reducing cerebral edema, improving neurological symptoms, and improving cognitive dysfunction after cerebral ischemia or stroke. The treatment refers to achieving a clinically significant area of cerebral infarction, reducing brain edema, improving neurological symptoms, and improving cognitive dysfunction after cerebral ischemia or stroke.

More specifically, the disease or symptom caused by preventing, alleviating and/or treating cerebral ischemia and/or cerebral ischemia according to the present invention is preventing, alleviating and/or treating a brain caused by global cerebral ischemia-reperfusion injury. Ischemia reduces the area of cerebral infarction in the type of cerebral ischemia, reduces brain edema, improves neurological symptoms and triggers memory impairment.

Another aspect of the present invention relates to a pharmaceutical composition for preventing, alleviating and/or treating a disease or a symptom caused by cerebral ischemia and/or cerebral ischemia, which comprises a prophylactically or therapeutically effective amount of ginsenoside Rgl, And optionally a pharmaceutically acceptable carrier and/or adjuvant. In the present invention, the ginsenoside Rgl pharmaceutical composition may be in a dosage form selected from the group consisting of: a solution, a suspension, an emulsion, a pill, a capsule, a powder, depending on the route of administration. Controlled release or sustained release formulation.

The ginsenoside Rgl pharmaceutical composition of the present invention can be formulated in a known manner and administered to a subject using several routes including, but not limited to, parenteral, oral, topical, intradermal, intramuscular, intraperitoneal, subcutaneous, Intranasal route.

The ginsenoside Rgl in the present invention can be produced by a commercially available or known method. The ginsenoside Rgl pharmaceutical composition of the present invention may optionally be formulated by any conventional method using one or more pharmaceutically acceptable carriers and/or excipients. Therefore, ginsenoside Rgl and pharmaceutically acceptable salts and solvates thereof can be specifically formulated, for example, by inhalation or insufflation (either through the mouth or the nose) or by oral, gastrointestinal, parenteral or rectal administration. Ginsenoside Rgl can take the form of a charged, neutral and/or other pharmaceutically acceptable salt. Examples of pharmaceutically acceptable carriers include, but are not limited to, those described in REMINGTON'S PHARMACEUTICAL SCIENCES (A. R. Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000).

The ginsenoside Rgl pharmaceutical composition may also be in the form of a solution, suspension, emulsion, pill, capsule, powder, controlled release or sustained release formulation. These formulations will contain a therapeutically effective amount of ginsenoside Rgl, preferably in purified form, together with a suitable amount of carrier to provide a form suitable for administration to a patient. The formulation should suit the mode of administration.

In the present invention, the purified form of ginsenoside Rgl means substantially pure ginsenoside Rgl, especially having a purity greater than 80%, preferably greater than 85%, particularly preferably greater than 90%, even more preferably greater than 95% Ginsenoside Rgl. Specifically, the purified form of ginsenoside Rgl may have a purity in the range of, for example, 90% to 96%. Parenteral administration

The ginsenoside Rgl pharmaceutical composition can be formulated for parenteral administration by injection, for example by bolus injection. Formulations for injection can be presented in unit dosage form in ampoules or in multi-dose containers containing optional preservatives. The parenteral preparation can be contained in an ampoule, disposable syringe or multi-dose bottle made of glass, plastic or the like. Preparation can It may be in the form of a suspension, solution or emulsion such as in an oily or aqueous vehicle, and may contain adjuvants such as suspending, stabilizing and/or dispersing agents.

For example, the parenteral preparation can be a sterile injectable solution or suspension (e.g., a solution in 1, 3-butanediol) in a non-toxic parenterally acceptable diluent or solvent. Acceptable carriers and solvents which may be employed include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspension medium. For this purpose any bland fixed oil may be employed including synthetic mono- and diglycerides. In addition, fatty acids such as oleic acid can also be used in parenteral formulations.

In addition, the ginsenoside Rgl pharmaceutical composition may also be formulated in powder form for reconstitution with a suitable carrier such as pyrogen-free sterile water prior to use. For example, a ginsenoside Rgl pharmaceutical composition suitable for parenteral administration may comprise a sterile isotonic saline solution containing from 0.1% to 90% by weight of ginsenoside Rgl per volume. For example, the solution may contain from about 5% to about 20%, more preferably from about 5% to about 17%, more preferably from about 8% to about 14%, still more preferably about 10%, of ginsenoside Rgl. The solution or powder formulation may also contain a solubilizing agent and a local anesthetic such as lidocaine to relieve pain at the injection site. Methods of administration of other parenteral compounds are known in the art and are within the scope of the invention.

Oral administration

For oral administration, the ginsenoside Rgl pharmaceutical composition can be used in the form of a tablet or a gelatin which is prepared by a conventional method using pharmaceutically acceptable excipients such as a binder, a filler, a lubricant and a disintegrating agent.

A. Adhesive

Binders include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, pistachio powder, guar gum, cellulose And derivatives thereof (for example, ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose), polyvinylpyrrolidone, mercapto cellulose, pregelatinized starch, hydroxypropyl fluorenyl Cellulose (for example, Nos. 2208, 2906, 2910), microcrystalline cellulose and mixtures thereof. Suitable forms of microcrystalline cellulose include, for example, materials sold as AVICEL-PH-101, AVICEL-PH-103, and AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pennsylvania). , USA). An example of a suitable binder is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose sold by FMC Corporation as AVICEL RC-581.

B. Filler

Fillers include, but are not limited to, talc, calcium carbonate (eg, granules or powders), lactose, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, Pregelatinized starch and mixtures thereof.

C. Lubricant

Lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate , talc, hydrogenated vegetable oil (for example, peanut oil, cottonseed oil, hollyhock oil, sesame oil, olive oil, corn oil and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar and mixtures thereof. Additional lubricants include, for example, solid silica gel (AER0SIL 200, manufactured by WR Grace Co. of Baltimore, Maryland, USA), condensed aerosols of synthetic silicon (sold by Deaussa Co. of Piano, Texas, USA), CAB-O- SIL (a pyrogenic silica product sold by Cabot Co. of Boston, Massachusetts, USA) and mixtures thereof.

D. Disintegrant

Disintegrators include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polycline potassium, sodium starch glycolate, potato or tapioca Starch, other starches, pregelatinized starch, other starches, clays, other alginates, other celluloses, gums, and mixtures thereof.

The tablets or capsules can optionally be coated by methods well known in the art. If the ginsenoside Rgl pharmaceutical composition of the present invention uses a binder and/or a filler, they are generally Formulated from about 50% to about 99% by weight of the compound. In one aspect, from about 0.5% to about 15% by weight of the disintegrant, especially from about 1% to about 5% by weight of the disintegrant, can be used in combination with ginsenoside Rgl. Lubricants may optionally be added in an amount generally less than about 1% by weight of ginsenoside Rgl. Techniques for preparing solid oral dosage forms and pharmaceutically acceptable tinting agents are described in Marsha ll, SOLID ORAL DOSAGE FORMS, Modern Pharmaceut i cs (Banker and Rhodes, Eds.), 7: 359-427 (1979). Other less typical formulations are well known in the art.

Liquid preparations for oral administration can be in the form of solutions, syrups or suspensions. Alternatively, the liquid formulation may be in the form of a thousand preparations which are reconstituted with water or a suitable carrier prior to use. These liquid preparations can be exemplified by conventional methods using pharmaceutically acceptable additives such as suspending agents (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats), emulsifiers (for example, lecithin or gum arabic), non-aqueous carriers (for example). , almonds, oily esters, ethanol or fractionated vegetable oils), and/or preservatives (for example, mercapto or propyl p-hydroxybenzoate or sorbic acid) are prepared. These formulations may also contain suitable buffer salts, flavoring, coloring, perfuming, and sweetening agents. Formulations for oral administration can also be formulated to effect controlled release of the compound. The oral preparation preferably contains 10% to 95% of the compound. Further, the ginsenoside Rgl pharmaceutical composition of the present invention can also be formulated into a tablet or lozenge for administration by a conventional method. Other methods of oral administration of ginsenoside Rgl pharmaceutical compositions are well known to those skilled in the art and are within the scope of the invention.

Controlled release administration

In order to prolong the activity of ginsenoside Rgl and reduce the frequency of administration, a controlled release (or sustained release) formulation can be formulated. Controlled release formulations can also be used to affect the onset time or other characteristics, such as blood compound levels, thereby affecting the production of side effects.

The controlled release formulation can be designed to initially release an amount of ginsenoside Rgl that produces the desired therapeutic effect, gradually and continuously releasing additional levels of ginsenoside Rgl to maintain a therapeutic level over a prolonged period of time. In order to maintain a nearly constant level of compound in the body, ginsenoside Rgl can be released from the dosage form at a certain rate instead of Ginsenoside Rgl metabolized and/or secreted in the body. The controlled release of ginsenoside Rgl can be stimulated by a variety of inducing factors, such as pH changes, temperature changes, enzymes, water, or other physiological conditions or molecules.

The controlled release system can include, for example, an infusion pump that can be used to administer the compound in a manner similar to delivering insulin or a chemotherapeutic agent to a particular organ or tumor. Using this system, ginsenoside Rgl is typically administered in combination with a biodegradable, biocompatible polymeric implant that releases ginsenoside Rgl at a selected location for a controlled period of time. Examples of polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyvinyl vinyl acetate, and copolymers and combinations thereof. In addition, the controlled release system can be placed near the treatment target, so only a portion of the systemic dose is required.

The ginsenoside Rgl of the present invention can be administered by other controlled release methods or drug delivery devices known to those skilled in the art. Including, for example, hydroxypropyl fluorenyl cellulose, other polymeric matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, etc., or any combination of the above, different The ratio provides the desired release profile. Other controlled release administration methods for ginsenoside Rgl are well known to those skilled in the art and are within the scope of the invention.

Inhalation administration

Ginsenoside Rgl can also be administered directly to the lungs by inhalation. For administration by inhalation, ginsenoside Rg 1 can be conveniently delivered to the lungs by a number of different devices. For example, metered-dose inhalers ("MDI"), which use tanks containing suitable low-boiling propellants, such as dichlorodifluorodecane, trichlorofluorodecane, dichlorotetrafluoroethane, carbon dioxide or other suitable Gas can be used to deliver compounds directly to the lungs. MDI devices are available from a number of suppliers such as 3M Corporation, Avent i s, Boehr inger Ing lehe im, Fores t Laboratories ^ Glaxo-We l lcome, Scher ing Plough and Vectura.

In addition, the compound can also be administered to the lungs using a dry powder inhaler (DPI) device. DPI devices typically use a mechanism, such as an explosive gas, to create a cloud in the container. Dry powder can then be inhaled by the patient. DPI devices are also well known in the art and are available from a number of suppliers including, for example, Fisons, Glaxo-Wel lcome, Inha le Therape ic Sys tems, ML Laboratories ^ Qdose and Vectura. One popular variation is the multi-dose DPI ("MDDPI") system, which allows for the administration of more than one therapeutic dose. MDDPI devices are available from companies such as As traZeneca, GlaxoWel lcome, IVAX, Schering Plough, SkyePharma and Vectura. For example, gelatin capsules and cartridges for use in inhalers and insufflators can be formulated as a powder mix containing ginsenoside Rgl and a powder base such as lactose or starch suitable for use in the system.

Another type of device that can be used to administer a compound to the lung is, for example, a liquid spray device provided by Aradigm Corporation. Liquid spray systems use tiny nozzles to atomize liquid compounds that can then be inhaled directly into the lungs. For example, a compound can be administered to the lungs using a nebulizer device. The atomizer generates an aerosol from the liquid compound preparation, for example, by using ultrasonic energy, to form fine particles which are easily inhaled. Examples of nebulizers include those provided by Sheff ie ld/Sys temic Pulmonary De l ivery Ltd., Avent i s and Batel le Pulmonary Therapeutics.

In another embodiment, an electrohydrodynamic ("EHD") aerosol device can be used to administer a compound to the lung. The EHD aerosol device uses electrical energy to atomize a liquid compound solution or suspension. When the compound is administered to the lungs using an EHD aerosol device, the electrochemical properties of the compound formulation are important parameters to be optimized. This optimization is routinely performed by those skilled in the art. Other intrapulmonary administration methods for ginsenoside Rgl are well known to those skilled in the art and are within the scope of the invention.

Liquid ginsenoside Rgl formulations suitable for use in nebulizers and liquid spray devices and EHD aerosol devices typically comprise ginsenoside Rgl and a pharmaceutically acceptable carrier. In a representative embodiment, the pharmaceutically acceptable carrier is a liquid such as an alcohol, water, polyethylene glycol or perfluorocarbon. Optionally, another substance may be added to alter the aerosol properties of the compound solution or suspension. For example, the substance can be a liquid, such as an alcohol, Glycol, polyethylene glycol or fatty acid. Other methods of formulating liquid compound solutions or suspensions suitable for use in aerosol devices are well known to those skilled in the art.

Depot administration

Ginsenoside Rgl can also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, the compound can be formulated with suitable polymeric or hydrophobic materials, such as emulsions in acceptable oils or ion exchange resins, or as sparingly soluble derivatives such as sparingly soluble salts. Other depot delivery methods for ginsenoside Rgl are well known to those skilled in the art and are within the scope of the invention.

Topical administration

For topical administration, ginsenoside Rgl can be combined with a carrier to deliver an effective dose, for example, from 1.0 μM to 1. O raM, depending on the desired activity. In one aspect of the invention, the topically administered pharmaceutical composition can be applied to the skin. The carrier can be, for example but not limited to, in the form of an ointment, cream, gel, paste, foam, aerosol, suppository, pad or gelling stick.

Topical formulations may also contain a therapeutically effective amount in ophthalmologically acceptable excipients such as buffered saline, mineral oil, vegetable oils such as corn oil or peanut oil, petrolatum, Mig lyol 182, alcohol solutions or liposomes or liposome-like products. compound of. Any of these compounds may also contain preservatives, antioxidants, antibiotics, immunosuppressants, and other biologically or pharmaceutically effective agents that have no deleterious effects on the compound. Other methods of topical administration of ginsenoside Rgl are well known to those skilled in the art and are within the scope of the invention.

Other drug delivery systems

Various other delivery systems are known in the art and can be used to administer the compounds of the invention. Moreover, these and other delivery systems can be combined and/or altered to optimize the administration of the ginsenoside Rgl of the present invention.

Another aspect of the invention also relates to a food product comprising ginsenoside Rgl. A further aspect of the invention relates to a method of preventing, ameliorating and/or treating a disease or condition caused by cerebral ischemia and/or cerebral ischemia, comprising administering to a patient in need thereof a therapeutically effective amount of ginsenoside Rgl or containing a therapeutically effective amount A quantity of a pharmaceutical composition of ginsenoside Rgl.

In the present invention, the therapeutically effective amount of ginsenoside Rgl may be any amount between 0.5 and 200 mg/kg body weight, preferably 1 to 150 mg/kg body weight, more preferably 2 to 100 mg/ The kg body weight is more preferably from 3 to 50 mg/kg body weight, more preferably from 4 to 35 mg/kg body weight, and even more preferably from 5 to 20 mg/kg body weight.

Principle for determining the effective dose of treatment

The toxicity and efficacy of ginsenoside Rgl can be determined by measuring LD _5D (50% population lethal dose) and ED ₅ in cell culture or experimental animals. A standard pharmaceutical method (50% effective therapeutic dose) is determined. The dose ratio between toxicity and efficacy is a therapeutic index and can be expressed as the ratio LD ₅ . /ED ₅ . . .

Data obtained from cell culture assays and animal studies can be used in dosage ranges formulated for use in humans and other mammals. Ginsenoside Rgl dose preferably have little or no toxicity includes ED _5. The circulating plasma or other body fluid concentration range.

The dosage may vary within this range, depending on the dosage form employed and the route of administration. For the ginsenoside Rgl of the present invention, the therapeutically effective dose can be estimated initially from cell culture assays. In animal models can be designed doses, to achieve the IC ₅ comprises as determined in cell culture. Circulating plasma concentration range (concentration of test compound to achieve half maximal symptom suppression). This information can be used to more accurately determine the doses that are useful in humans and other mammals. The ginsenoside Rgl level in plasma can be determined, for example, by high performance liquid chromatography.

The amount of ginsenoside Rgl which can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. It will be understood by those skilled in the art that the unit content of ginsenoside Rgl contained in individual doses of each dosage form is not The need to constitute a therapeutically effective amount by itself can be achieved by administering multiple individual doses to achieve the desired therapeutically effective amount. The choice of dosage will depend on the dosage form employed, the condition being treated, and the particular purpose to be achieved according to the discretion of those skilled in the art.

The dosage regimen for treating a disease or condition with the ginsenoside Rgl of the present invention is selected according to a variety of factors, including the type of patient, age, weight, sex, diet and medical condition, route of administration, pharmacological considerations, such as activity, effectiveness The pharmacokinetic and toxicological profile of the particular compound used, whether or not the ginsenoside Rgl drug delivery system is used. Therefore, the dosage regimen actually used may be very different between the subject and the subject. DRAWINGS

Figure 1 Effect of transient global cerebral ischemia and ginsenoside Rgl on the incubation period of Mongolian gerbils. *P < 0.05 for the cerebral ischemia group, **P < 0.01 for the cerebral ischemia group, n=8 (student t-test).

Figure 2 The effect of transient global cerebral ischemia and ginsenoside Rgl on the number of errors in the darkness of Mongolian gerbils. *Ρ<0· 05 For the cerebral ischemia group, **Ρ<0.01 for the cerebral ischemia group, η=8.

Figure 3. Effect of ginsenoside Rgl on the escape latency in the Morris water maze experiment. *P < 0.05 for the cerebral ischemia group, (one-way AN0VA, followed by post hoc tests). detailed description

The following examples specifically show the application of the present invention. However, the present embodiment does not limit the scope of use of the present invention. Example 1 Anti-cerebral ischemia effect of ginsenoside Rgl

I. Effect of ginsenoside Rgl on brain damage in rats with middle cerebral artery occlusion and reperfusion Objective: To observe the effect of ginsenoside Rg l on brain damage in rats with middle cerebral artery occlusion and reperfusion.

1. Materials and methods

1. 1 Experimental animals: Male Wis tar rats weighing 280-320 g were provided by the Institute of Zoology, Chinese Academy of Medical Sciences. Animals were free to eat, and the environment was adapted to the environment in a clean animal room before the test.

1. 2 reagents and preparation:

Ginsenoside Rgl (Bethune Medical University) is formulated in distilled water to prepare 5. Omg/kg, 10 mg/kg, 20 mg/kg (5 ml/kg body weight) daily for intragastric administration.

1. 3 methods:

Middle cerebral artery occlusion reperfusion: anesthetized with chloral hydrate (350 mg/kg, ip), 2 cm mid-neck incision, exposed common carotid artery (CCA) and its branching external carotid artery (ECA) and neck on the left side Internal artery (ICA). The branch occipital artery of ECA, the superior sacral artery and the ECA terminal branch were ligated to separate the extracranial branch pterygopalatine artery of ICA. Clamp the CCA, ICA, insert the nylon thread with a diameter of 0.3 mm from the ECA, introduce the wire into the ICA, loosen the ICA clip, and continue to insert the nylon thread into the intracranial segment. The length of the inserted nylon thread is about 2 cm. Block the middle cerebral artery (MCA). After 2 hours, the line is pulled out, the ECA stump is ligated, and the CCA is released.

Male Wis tar rats weighing 280-320 g were randomly divided into five groups: MCA0 model group, 5, 10, 20 mg/kg Rgl group, and sham operation group, 8 animals per group. Rgl was administered intragastrically daily for 24 hours before surgery, 0.5 hours, 24 hours after surgery, and 46 hours from the end of the trial. Distilled water (5 ml / kg body weight) was administered intragastrically at the same time in the MCA0 model group and the sham operation group. Among them, the sham operation group performed certain surgical operations, but did not block the middle cerebral artery; the model group animals blocked the middle cerebral artery and caused focal cerebral ischemia.

1. 4 Behavioral Score:

(1) Lift the rat tail off the ground and observe the forelimbs. The forelimb of normal rats symmetrically extends to the ground. If the right shoulder is internal rotation, the right forelimb is the recipient, which is rated 1-4 points, otherwise 0 points.

(2) Place the animal on the smooth floor and push the left shoulder to the opposite side. Depending on the degree of drag reduction, it is rated 1-3 points.

(3) Place the animal on the metal net, gently pick up the rat, and the muscle of the right forelimb is decreased, and the score is 0-3 according to the degree.

The above standard scores, out of 10 points, the higher the score, the more serious the animal's behavioral disorder.

1. 5 Measurement of infarct size:

Animals took the brain, removed the olfactory bulb, cerebellum and low brainstem, cut into five pieces, stained with 4% red tetrazolium, protected from light, and incubated at 37 °C for 30 min. The infarct size was white, and the infarcted brain tissue and whole brain weight were weighed, and the infarct size was determined as a percentage.

1. 6 Determination of water content:

The amount of water content represents the extent of cerebral edema. The brain tissue was dried at 120 °C for 48 hours, and the water content of the table was (the ratio of brain wet weight to brain dry weight)/brain wet weight. High water content, severe brain edema.

2. Results statistics:

分析 Analyze with T test. See Table 1 for the results.

*P<0. 05 **P<0. 01 3. Conclusion:

A variety of different concentrations of ginsenoside Rgl can effectively improve the behavioral score, reduce the area of cerebral infarction, reduce brain edema, and have obvious anti-cerebral ischemia. Example 2 The improvement of memory impairment caused by cerebral ischemia

Experimental methods: Male Mongolian gerbils, weighing 66-80 g, were purchased from the Experimental Animal Center of the Capital Medical University. The purity of ginsenoside Rgl monomer was greater than 98%. It was purchased from the Organic Chemistry Laboratory of Bethune Medical University.

Ginsenoside Rgl was prepared with 0.9% physiological saline, JZZ94 passive avoidance recorder,

Morr i s water maze automatic monitor, manufactured by the Instrument Research Laboratory, Institute of Materia Medica, Chinese Academy of Medical Sciences.

First, gerbil passive avoidance ability test

Darkness behavior experiment: The experimental device consists of two chambers of the same size, with a volume of 1 8 x 2 5 x 2 0. A 1 5W fluorescent lamp is suspended above the bright room. The bottom of the darkroom consists of a stainless steel grid with a spacing of 1. 1 cm. It is connected to a voltage of 40 volts. There is a circular hole with a diameter of 3 cm between the two chambers. On the third day after ischemia, the gerbils were placed in the monitoring box for 5 minutes to familiarize them with the surrounding environment. The next day, the memory was trained. The electric shock switch was turned on, and the gerbils were placed in the bright room, and the gerbils were darkened. Habits enter the darkroom, and after being shocked, they quickly escape from the darkroom. The experimental conditions were controlled to ensure that each animal received only one electric shock before being removed. After 24 hours, the gerbils were placed in the bright room again, and the computer automatically recorded the incubation period and number of entries of the gerbils into the darkroom within 80 seconds. Second, gerbil space learning ability test

The modified Morr is water maze consists mainly of a metal cylindrical basin (60 cm high, 80 cm diameter), automatic detection, display, recording device and safety island. All activities of the animals in the pool can be tracked and monitored through the monitor, and the input computer can be processed and analyzed by special software. The animal found a safe island in 60 seconds. The actual time is counted as the incubation period; if not found, 60 seconds is used as the incubation period. Generally speaking, the activity track of animals in the pool can be divided into four types: edge type, random type, trend type and straight type. The edge type means that the animal has no intellectual activity, and only mechanically searches for the exit along the labyrinth wall. There is no concept of a safe island, which is the worst way to learn memory. Random type means that the animal has the concept of a safe island, but does not remember the specific location and randomizes Looking for; the trend refers to the concept that the animal not only has a safe island, but also remembers the approximate location of the safe island and looks for it purposefully; the straight line is the best performance of the animal's ability to learn and remember, and the animal is accurately remembering the location of the safe island. Find the track as a straight line. In the experiment, in addition to recording the incubation period of the animal looking for a safe island and finding the trajectory, the length of the route for finding the path was also recorded.

Before the start of the experiment, a special transparent safety island was placed in the first quadrant of the Morr i s water maze, and two different visual cue markers were placed on the inner wall of the quadrant, the height of which was determined by the gerbils. After the start of the formal experiment, the degreased instant milk powder was dissolved in warm water, and the drum was filled with milky white opaque liquid in order to interfere with the animal's line of sight, making it impossible for them to see the underwater safe island with their eyes. The water temperature was required to be controlled at 22 ± 2 during the experiment. C, the water surface is about 1 cm above the safety island. Above the pool, the camera is connected to the monitor screen and the computer to automatically monitor and record the entire activity track of the gerbil and its arrival time on the safe island. The computer records, processes and calculates the data synchronously.

On the 28th day after the operation, the water depth in the drum was controlled to about 6 cm, and the animals were trained to be familiar with the test environment. Formal training begins the following day: 2 times a day, with three or four quadrants as the entry point, each gerbil is trained for 1 minute each time. After the training, the animals were placed on a safe island for 20 seconds to achieve enhanced memory. The experimental results are expressed in terms of the latency of the gerbils looking for a safe island, the length of the swimming route, and the strategy used. The effect of transient global cerebral ischemia on passive evasive memory ability of gerbils and the protective effect of Rgl

Capturing Mongolian gerbils bilateral common carotid artery 6m in causing transient global cerebral ischemia At the beginning of 4 hr, ginsenoside Rgl or the same volume of normal saline was given intraperitoneally daily, 23 days (at this time, the animal had no obvious motor dysfunction), and the passive avoidance memory ability of the animals was tested by the automatic avoidance monitoring device. The results showed that the latency of the gerbils entering the dark room and the number of errors entering the electric shock zone were significantly shorter than those of the sham-operated group (P < 0.05, P < 0.01), indicating transient global cerebral ischemia. It can indeed cause the animal to passively avoid the damage of memory ability. The specific manifestation is that the gerbil is quickly forgotten after being shocked and enters the electric shock chamber multiple times in a short time. After the continuous administration of ginsenoside Rgl, the incubation period of the gerbil into the electric shock zone is prolonged. The number of errors also showed a certain degree of reduction (Fig. 1, 2, ), especially in the Rgl (5mg/kg) group. The above two effects were significantly different from the model group (latency: P < 0.01, P < 0. 05 ) The results of this experiment suggest that Rgl can successfully improve the passive avoidance memory ability caused by transient global cerebral ischemia. IV. The effect of transient global cerebral ischemia on spatial learning and memory ability of gerbils and ginsenosides The role of Rgl.

Two days after the end of the darkness test, we used Morr i s water maze to observe the spatial learning and memory ability of each group of gerbils. At this time, the global cerebral ischemia occurred for nearly 5 weeks. There was no obvious neurological deficit in each group of gerbils. At the same time, there was no significant difference in swimming speed during training. Therefore, the motor dysfunction itself can be excluded from the space recognition. The influence of knowledge.

After 4 days of cognitive training, we found that the latency of each group of gerbils looking for a safe island was gradually shortened, and the search strategy was also transitioned from the initial edge and random to the trend and straight line. Memory of the location of the safe island can be formed by spatial cues in the surrounding environment. As shown in Figure 3, compared with sham surgery, the ischaemia period of the ischemic group was prolonged, and the search strategy was more moderate from the edge and random to the trend and the linear. But the above changes are just a trend, lacking statistical significance, saying Transient global cerebral ischemia at 6 min may not cause a significant reduction in spatial learning and memory in animals. Compared with the ischemic group, the long-term administration of ginsenoside Rgl is shorter than the ischemic group. The latency of finding a safe island is shortened, and the distance is reduced. As the number of training increases, the purpose of the animal's search strategy is also strong. And the linear strategy has a larger ratio). Especially in the 5mg/kg group, the incubation period was significantly shorter than that of the ischemic group on the second day of training (P<0.05). The four-day swimming distance was also significantly shorter than the simple ischemia group (P<0.05), suggesting that Rgl may be The spatial cognitive ability of gerbils plays a certain role.

Claims

Rights request

1. Use of ginsenoside Rgl for the preparation of a medicament for preventing, alleviating and/or treating a disease or condition caused by cerebral ischemia and/or cerebral ischemia.

2. The use according to claim 1, wherein the disease or symptom caused by preventing, alleviating and/or treating cerebral ischemia and/or cerebral ischemia is selected from the group consisting of reducing infarct size, reducing brain edema, improving neurological symptoms, and improving recognition. Know dysfunction.

3. A pharmaceutical composition for preventing, alleviating and/or treating a disease or condition caused by cerebral ischemia and/or cerebral ischemia, comprising a prophylactically or therapeutically effective amount of ginsenoside Rgl, and optionally a pharmaceutically acceptable Accepted carrier and / or excipients.

The pharmaceutical composition according to claim 3, wherein the disease or symptom caused by preventing, alleviating and/or treating cerebral ischemia and/or cerebral ischemia is selected from the group consisting of reducing infarct size, reducing brain edema, improving neurological symptoms, and Improve cognitive dysfunction.

The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition may be in a dosage form selected from the group consisting of a solution, a suspension, an emulsion, a pill, a capsule, a powder, a controlled release or sustained release preparation.

6. A method of preventing, ameliorating and/or treating a disease or condition caused by cerebral ischemia and/or cerebral ischemia, comprising administering to a patient in need thereof a therapeutically effective amount of ginsenoside Rgl or a pharmaceutical combination according to claim 3. Things.

7. The method of claim 6, wherein the preventing or alleviating and/or treating a disease or symptom caused by cerebral ischemia and/or cerebral ischemia is selected from the group consisting of reducing infarct size and reducing brain Edema, improve neurological symptoms and improve cognitive dysfunction.

8. The method of claim 6, wherein the administration is by a route selected from the group consisting of, but not limited to, parenteral, oral, topical, intradermal, intramuscular, intraperitoneal, subcutaneous, intranasal.

9. The method of claim 6, wherein the therapeutically effective amount of ginsenoside Rg l is any amount between 0.5 and 200 mg/kg body weight, preferably 1-150 mg/kg body weight, more preferably Any amount between 2 - 100 mg / kg body weight, more preferably 3 - 50 mg / kg body weight, more preferably 4 - 35 mg / kg body weight, more preferably 5 - 20 mg / kg body weight

1 0. A food product containing ginsenoside Rgl.