JP2001302517A - Prophylactic and remedy for brain edema - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a prophylactic or a remedy for brain edema containing trehalose as an active ingredient. SOLUTION: The trehalose has excellent edema-inhibiting effects compared to mannitol, and high safety, and thereby is useful as the therapeutic agent of the brain edema caused by ischemia or another cerebral disorder. The trehalose can be used as the prophylactic agent of the brain edema by administering the trehalose before onset of the brain edema.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an agent for preventing or treating cerebral edema.

[0002]

2. Description of the Related Art In recent years, the mortality rate due to cerebrovascular disorders has been decreasing, but it is thought that the number of patients with cerebral infarction due to cerebral ischemia will further increase in the aging society.
Although cerebral infarction causes various pathological conditions depending on the site of onset, all of them cause serious symptoms such as speech disorder, limb movement disorder and memory disorder by inducing nerve cell death. The cause of nerve cell death due to such brain damage is that, as a result of the inhibition of the metabolic function of nerve cells, edema occurs in a narrow space surrounded by the skull, thereby compressing the tissue and inhibiting blood flow. It is considered. Since the brain is a tissue that requires particularly intense energy, even when small blood vessels are occluded, it is highly possible that locally generated edema inhibits blood flow to surrounding tissues and causes cell death. Therefore, there is a high clinical need for drugs that suppress cerebral edema.

Heretofore, hyperosmotic drugs such as mannitol have been exclusively used as drugs for suppressing brain edema. That is, mannitol is not metabolized in the living body and has no pharmacological action by itself, but is distributed only in the extracellular fluid. Therefore, mannitol is used for removing water and sodium ions stored in tissues by osmotic pressure.

[0004] However, mannitol requires a large amount of administration due to its nature, and its edema inhibitory effect is not always sufficient, and the life support and prognosis of patients are not satisfactory. Therefore, a drug having a higher edema inhibitory effect than mannitol was clinically desired, but a measurement system capable of objectively quantifying cerebral edema and screening for an anti-edema drug has not been established. At present, no drug has been found yet.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a drug for preventing or treating cerebral edema which has an excellent edema suppressing effect and is highly safe.

[0006]

Means for Solving the Problems The present inventors have searched for a substance that suppresses brain edema by using an objective analysis method of cerebral edema using an infrared image analyzer, and found that saccharides that replace sucrose It has been found that trehalose, which is used as a drug, has an excellent cerebral edema inhibitory effect and is useful as an agent for preventing or treating cerebral edema, and completed the present invention.

That is, the present invention provides a preventive or therapeutic agent for cerebral edema containing trehalose as an active ingredient.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION Trehalose, which is an active ingredient of the preventive or therapeutic agent for cerebral edema of the present invention, is a disaccharide in which two molecules of glucose are bonded to each other by a reducing group. In nature, bacteria, fungi, Widely distributed in algae, insects, etc.
In recent years, demand for trehalose has been rapidly increasing in the fields of foods and cosmetics because it has a water retention property as a saccharide replacing sucrose. However, in the field of pharmaceuticals, it is only reported that they have an effect of increasing bone mass.

Trehalose includes α, α-form, α, β-form and β, β-form due to the difference in the binding mode of two molecules of glucose.
There are three isomers of the body. In the present invention, any of these isomers can be used as long as they have a cerebral edema inhibitory effect, but are preferably α, α isomers.

Such trehalose can be prepared by a known method for any of the isomers. For example, α, α isomers include non-reducing saccharide-forming enzyme and trehalose-releasing enzyme in a partial hydrolyzate of starch. Can be prepared. Further, the obtained trehalose is preferably purified and used as necessary by, for example, a desalting means including a ion exchange resin, a porous resin, activated carbon and a membrane filter, an adsorption means and a filtration.

[0011] The cerebral edema inhibitory effect of such trehalose is
It can be confirmed by using an objective analysis method of cerebral edema using the infrared image analysis device found by the present inventors (see Examples). That is, as shown in Examples described later, a stimulus for inducing brain edema is given to a brain slice, a temporal change of the brain cell is obtained as infrared image data, and this is obtained by image analysis, and the infrared image is obtained. Average brightness value,
The method of quantitatively grasping the state of edema from the standard deviation value of the average luminance of the infrared image and the contrast value obtained from their ratio showed that trehalose has a better brain edema inhibitory effect than mannitol .

The preventive or therapeutic agent for cerebral edema of the present invention is aqueous,
It is preferably used in the form of a suspension or emulsion of an injection or a drop, but other general pharmaceutical preparations, that is, tablets, granules, fine granules, powders, capsules, syrups And elixirs can be used orally.

Injections are usually prepared by adding an effective amount of trehalose and optional ingredients such as stabilizers, local anesthetics and preservatives to water, and then dissolving the solution with a membrane filter or the like. After filtration, the container is appropriately filled aseptically and sealed as it is, or lyophilized and then sealed. In addition, such injection preparations may contain, if desired, components usually used in infusions other than trehalose, for example, glucose, maltose, fructose, sorbitol, saccharides such as xylitol, sodium chloride, sodium acetate, sodium lactate, and citric acid. Sodium, sodium monohydrogen phosphate, sodium dihydrogen phosphate, potassium chloride, potassium lactate, potassium citrate, potassium monohydrogen phosphate, potassium dihydrogen phosphate, calcium acetate, calcium lactate, calcium gluconate, magnesium sulfate, chloride Electrolytes such as magnesium,
Amino acids, vitamins, and drugs such as drugs for suppressing brain inflammation, antibiotics, and analgesics can also be added. When formulated as a freeze-dried preparation, it is possible to add a saccharide such as mannitol, inositol, lactose, maltose, sucrose, or a sugar alcohol or an excipient such as glycine, which is easily freeze-dried, if necessary. it can.

Formulation as internal preparations such as granules, tablets, capsules and the like includes excipients, binders, disintegrants, disintegration inhibitors,
Absorption enhancers, moisturizers, adsorbents, lubricants, coloring agents, manufactured using carriers for preparations such as fragrances, tablets, if necessary, tablets coated with normal skin, such as sugar-coated tablets, gelatin Encapsulated tablets, enteric-coated tablets, and film-coated tablets, and capsules are prepared by filling hard gelatin capsules, soft capsules, and the like.

The amount of trehalose to be incorporated in the preventive or therapeutic agent for cerebral edema of the present invention is appropriately selected according to the dosage form. In the case of an injectable preparation, the concentration of trehalose is 1% by weight or more. It is preferable that the compound is blended so as to be preferably 1.5 to 2% by weight.
It is preferable that the content is 10 to 15% by weight in the composition.

The method of administration of the agent for preventing or treating cerebral edema of the present invention is not particularly limited, and may be various preparation forms, patient age, sex,
Although the administration is carried out by a method according to the degree of the disease, administration by injection (intravenous, intramuscular, intradermal, subcutaneous or intraperitoneal) is preferred, and intravenous administration is particularly preferred. In addition, direct administration to the injured area of the skull can be expected to be even more effective.

The dose of the prophylactic or therapeutic agent for cerebral edema of the present invention can be appropriately selected depending on the usage, age of the patient, gender, degree of disease, etc., but is preferably 0.5 to 200 mg / day. Weight.

[0018]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Method for analyzing cerebral edema The outline of the method for analyzing cerebral edema used in the present invention will be described below. (1) A stimulus to induce cerebral edema is given to a brain slice, and a temporal change of the brain cell is obtained as infrared image data.
As the brain slice, a brain of an experimental animal such as a rat or a mouse, for example, a cerebral cortex, a cerebellum, or a hippocampus is used, which is sliced to a thickness of 300 to 400 μm by a vibrating tissue slicer or the like. Examples of the stimulus to induce cerebral edema include ischemia, electrical stimulation, and physical and chemical stimuli such as drugs.However, the ischemic state is prepared by perfusing a brain slice with a solution in which oxygen is aerated. It is performed by changing the perfusate to an oxygen-free and glucose-free solution. An infrared differential interference microscope is used exclusively for acquiring image data, and a brain section is photographed in real time by an infrared detection device (for example, an IR-CCD camera (manufactured by Hamamatsu Photonics KK)) attached to the infrared differential interference microscope. Then, the cell shape in the deep part of the brain is two-dimensionally infrared-imaged (see FIG. 1).

(2) The acquired infrared image data is image-processed by a general-purpose image processing device (for example, Argus 50 (manufactured by Hamamatsu Photonics)) (see FIG. 1). The average luminance value, the standard deviation value of the average luminance of the infrared image, and the contrast value are measured and calculated. Here, the average luminance value of the infrared image is a value obtained by digitizing the infrared image by an image processing apparatus and dividing the total sum of the luminance values in the entire image or a total range of pixels by the total number of pixels. The standard deviation is a standard deviation calculated for the average value. The contrast value is an index indicating the image quality of the infrared image calculated by the following equation from the average luminance of the infrared image and its standard deviation.

[0020]

(Equation 1)

(3) By such an analysis, it is possible to quantify the time course of the loss of contrast in an image due to an increase in the transmittance of infrared rays due to edema of cells. By analyzing the three parameters of the average luminance value of the infrared image, the standard deviation value of the average luminance and the contrast value, it is possible to quantify and understand the quality of ischemic edema and edema caused by other causes, Also, whether or not the above parameters have been recovered after releasing these stimuli coincides with the need to treat cerebral edema with these stimuli. Therefore, the test drug-containing solution is perfused after the ischemic state is released, and the cerebral edema inhibitory effect can be evaluated by analyzing the recovery of the three parameters over time.

Example 1 Effect of Trehalose on Ischemic Edema A solution (ischemic blood) prepared by removing enzymes and glucose added to artificial cerebrospinal fluid was prepared and exposed to a hippocampal slice specimen of a rat for 10 minutes. Created an ischemic condition. After ischemia, the artificial cerebrospinal fluid was perfused with 30 mM and 50 mM of trehalose and a solution in which no drug was added and 50 mM and 100 mM of mannitol as a comparison, and the cerebral spinal fluid was treated with an infrared image analyzer (FIG. 1). The changes were acquired as infrared image data (FIGS. 2-4). Further, the contrast value was obtained by the above equation, and the change was analyzed (FIGS. 5 to 7).

In a normal state before ischemia, the morphology of the cell body and dendrites extending from the cell body is clear, but the cell body swells 10 minutes after ischemia, and the structure of the dendrites changes. It is no longer visible. Even when perfused with a drug-free solution for 35 minutes or longer, no tendency for recovery was observed (FIG. 2). On the other hand, when perfused with a solution containing 30 mM trehalose, edema tended to recover (FIG. 3), and when perfused with a solution containing 50 mM trehalose, good recovery of edema was observed. (FIG. 4). The above-mentioned edema-suppressing effect was confirmed by a slight increase in the contrast value when 30 mM of trehalose was added, and almost complete recovery of the contrast value when 50 mM was added (FIGS. 5 and 7). On the other hand, the addition of mannitol
At the addition of 00 mM, an increase in the contrast value was observed, but at the addition of 50 mM, no increase in the contrast value was observed, but rather a decrease was observed (FIGS. 6 and 7). It was shown that the edema suppressing effect was high and the durability was high.

Example 2 Injection Trehalose, from which pyrogen had been previously removed, was dissolved in distilled water for injection to a concentration of 1.5% by weight, filtered through a membrane filter, filled into a container, and charged with high-pressure steam according to a conventional method. It was sterilized to make an injection.

[0025]

EFFECT OF THE INVENTION Trehalose has a superior edema inhibitory effect as compared with mannitol and is highly safe, so that it is useful as a therapeutic agent for cerebral edema caused by ischemia or other brain disorders. By administering before the onset of edema, it can be used as a drug for preventing brain edema.

[Brief description of the drawings]

FIG. 1 is a configuration diagram of an infrared differential interference contrast microscope-image processing apparatus.

FIG. 2 is a microscopic image (using a 40 × objective water immersion lens) of rat hippocampal CA1 pyramidal cell layer when perfused with a drug-free artificial cerebrospinal fluid after 10 minutes of ischemia. .

FIG. 3 shows rat hippocampus CA1 when perfused with trehalose (30 mM) solution after ischemia for 10 minutes.
Microscope image of pyramidal cell layer (40x objective water immersion lens)
It is.

FIG. 4 shows rat hippocampus CA1 when perfused with a trehalose (50 mM) solution after ischemia for 10 minutes.
Microscope image of pyramidal cell layer (40x objective water immersion lens)
It is.

FIG. 5 is a diagram showing the change over time of the contrast value when perfused with a trehalose (30 mM, 50 mM) solution after being in an ischemic state for 10 minutes.

FIG. 6 is a diagram showing the change over time of the contrast value when perfused with a mannitol (50 mM, 100 mM) solution after ischemic state for 10 minutes.

FIG. 7 shows a control group, a mannitol added group (50 mM, 100 mM) and a trehalose added group (3
FIG. 10 is a diagram showing average values (5 examples) of the time course of the contrast value at 0 mM and 50 mM).

Claims (1)

[Claims] 1. A preventive or therapeutic agent for cerebral edema comprising trehalose as an active ingredient.