CN103565857B - Use of water clove extract for preparing composition for enhancing memory function and delaying neurodegeneration - Google Patents

Abstract

The invention discloses application of a ludwigia octovalvis extract in preparing a composition for enhancing memory function and delaying neurodegeneration and application of the ludwigia octovalvis extract in preparing a composition for treating Alzheimer's disease. The water clove extract is used as an active ingredient, and a medical composition or a food composition for enhancing memory function and delaying neurodegeneration can be prepared for patients with Alzheimer's disease.

Description

Use of water clove extract for preparing composition for enhancing memory function and delaying neurodegeneration

technical field

The present invention relates to water clove (Ludwigiaoctovalvis) extract, specifically, the present invention relates to a kind of use of water clove extract for preparing the composition of enhancing memory function and delaying neurodegeneration.

Background technique

As the global population is aging, Alzheimer's disease has also become a problem faced by many countries. According to research estimates, more than 24 million people around the world suffer from this disease. And when the condition of Alzheimer's disease becomes more and more serious, the patient needs the help of others in all aspects of life, such as washing, eating and going to the toilet. Because people with Alzheimer's disease need people to take care of them day and night, the lives of their relatives and friends are often greatly affected.

There is still no accurate diagnosis and effective treatment for Alzheimer's disease, so scientists all over the world are devoting themselves to the research and development of new drugs or treatments.

Therefore, there is still a need for a pharmaceutical or food composition that enhances memory function and delays neurodegeneration.

Contents of the invention

The present invention provides a novel application of water clove extract in preparing a composition for enhancing memory function and delaying neurodegeneration.

The invention proposes the use of a water clove extract for preparing a composition for enhancing memory function and delaying neurodegeneration.

The invention also proposes the application of the water clove extract for preparing a composition for treating Alzheimer's disease.

In one aspect, the present invention provides a composition for enhancing memory function and delaying neurodegeneration, which comprises an effective amount of water clove (Ludwigiaoctovalvis) extract.

The invention provides a composition for treating Alzheimer's disease, which comprises an effective amount of water clove extract.

According to the present invention, the water syringa extract can be a water syringa water extract or a water syringa alcohol extract.

The composition for enhancing memory function and delaying neurodegeneration of the present invention can be a pharmaceutical composition or a food composition.

Moreover, the composition is a pharmaceutical composition, and contains an effective amount of clove extract and a pharmaceutically acceptable carrier.

In a specific embodiment of the present invention, the clove extract is prepared by a method comprising the following steps: (a) extracting clove with a working liquid to obtain clove extract and residue; (b) separating the residue and (c) drying the water clove extract by a drying process to obtain the water clove extract.

Wherein, a step (d) is further included after the step (c), redissolving the water clove extract with the working liquid, and the concentration of the water clove extract is between 0.005% and 0.5%.

Wherein, in the step (a), the working liquid is used to extract the clove for 4-12 hours.

Wherein, the temperature of the working liquid is 2-10°C.

Wherein, in the step (a), after extracting cloves with the working liquid for 4-12 hours, heat it for 10-60 minutes.

Wherein, a step (b1) is further included after the step (b), which immerses the separated clove residue in another extraction working liquid, and heats and filters the extraction working liquid to produce a water clove secondary extract, and then dissolve the water clove secondary extract into the water clove extract.

Wherein, the heating temperature is 50-100°C.

Wherein, the water clove extract is the water clove ethanol extract.

By adopting the present invention, the extract of cloves can be used as the active ingredient, combined with known techniques in the art, to prepare a pharmaceutical composition or a food composition that enhances memory function and delays neurodegeneration for use by Alzheimer's patients.

Without further elaboration, the applicant believes that one skilled in the art can, based on the preceding description, utilize the present invention to its widest extent. Therefore, it can be understood that the following descriptions are only used for illustration and not to limit the rest of the disclosure in any way.

Description of drawings

Figure 1 shows the relationship between the concentration of the water clove extract and the learning and memory of Drosophila;

Figure 2 shows the relationship between the concentration of clove extract and the memory function of mice (object cognition experiment); and

Figure 3 shows the relationship between the concentration of clove extract and the memory function of mice (elevated T-maze test).

detailed description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The word "a" used herein, unless otherwise specified, refers to at least one (one or more) quantity.

The invention discloses the novel use of the water clove extract in preparing a composition for enhancing memory function and delaying neurodegeneration and the application of the water clove extract in preparing a composition for treating Alzheimer's disease. Those skilled in the art can use the extract of Ludwigiaoctovalvis to enhance the memory function and delay neurodegeneration, use the extract of Ludwigiaoctovalvis as the active ingredient, and cooperate with the known technology in the field to prepare a drug that enhances memory function and delays neurodegeneration. The pharmaceutical composition or food composition is used by Alzheimer's patients.

According to the present invention, the water syringa extract can be a water syringa water extract or a water syringe alcohol extract, which can be obtained by extracting natural or herbal medicines known in the art. For example, water clove can be immersed in a working liquid for extraction, and the working liquid is preferably a non-toxic and stable solvent. In a specific embodiment of the present invention, the citrus water extract is an ethanol extract.

In a specific embodiment of the present invention, the clove extract is prepared by a method comprising the following steps: (a) extracting clove (Ludwigiaoctovalvis) with a working liquid to obtain clove extract and residue; (b) separating the residue; and (c) drying the clove extract by a drying process to obtain the clove extract. In another specific embodiment, the method further includes: (d) dissolving the clove extract with the working liquid so that the concentration of the clove extract is between 0.005% and 0.5%. According to a specific embodiment of the present invention, in the step (a), the working liquid is used to extract the clove for 4-12 hours, wherein the temperature of the working liquid is 2-10°C. In another specific embodiment, after the step (a) extracts cloves with the working liquid for 4-12 hours, it is further heated for 10-60 minutes. In addition, in the step (a), the water clove can be ground before extraction. In some specific embodiments of the present invention, after step (a) soaking cloves in the working liquid for extraction for a period of time, the extraction working liquid is further heated to 50-100° C. for further extraction. When the working liquid is alcohol, the water heating can be used. In a specific embodiment of the present invention, step (b) is to separate the residue by centrifugal filtration. According to the present invention, the drying process in step (c) can use methods known in the art, for example, freeze drying or concentration under reduced pressure. According to a specific embodiment of the present invention, after step (b), it further includes: (b1) extracting the residue with a working liquid, heating and filtering to obtain a secondary extract of cloves, and then cloves two The secondary extract is dissolved in the water clove extract.

The invention provides a composition for enhancing memory function and delaying neurodegeneration, which comprises an effective amount of water clove (Ludwigiaoctovalvis) extract. The present invention also provides a composition for treating Alzheimer's disease, which comprises an effective amount of water clove extract.

The term "effective amount" as used herein refers to the amount of clove extract that has the effect of enhancing memory function and delaying neurodegeneration for the subject, whether it is administered alone or co-administered with other active substances. As known to those skilled in the art, the therapeutically effective amount will vary with the route of administration, the amount of excipient used, and co-administration with other active substances.

The term "treating" as used herein refers to curing, alleviating, alleviating, altering, ameliorating a disease, symptoms of a disease or arresting the progression of a disease.

The term "delaying neurodegeneration" as used herein refers to alleviating, alleviating or improving the symptoms of neurodegeneration, including but not limited to the reduction or decline of memory function, for example, alleviating the reduction of memory function caused by Alzheimer's disease or decline.

The composition of the present invention can be a pharmaceutical composition or a food composition. The pharmaceutical composition or food composition of the present invention can be prepared by mixing the hydrangea extract with existing carriers, diluents, solvents, coloring agents, antioxidants, inert substances, etc. and/or other additives formulated into tablets, capsules, powders, granules, infusions, beverages, health foods or food additives.

Compositions for oral administration may be in any form suitable for oral administration such as, but not limited to, solutions, suspensions, lozenges, pills, capsules or powders. In the case of lozenges, known excipients such as lactose or corn starch may be included. Lubricants, such as magnesium stearate, are also commonly added. In capsule form, diluents that may be used include lactose or corn starch. In the form of aqueous solution or emulsifier, the active ingredients can be suspended, emulsified or dissolved in the oil layer. Sweetening, flavoring or coloring agents may further optionally be added.

In one aspect, the present invention provides a pharmaceutical composition for enhancing memory function and delaying neurodegeneration, which comprises an effective amount of water clove (Ludwigiaoctovalvis) extract.

The pharmaceutical composition may additionally contain a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein refers to any known type of non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material, formulation adjuvant or vehicle. A pharmaceutically acceptable carrier is nontoxic to recipients and compatible with the other ingredients of the formulation at the dosages and concentrations employed. Pharmaceutically acceptable carriers are generally readily available to the public. In addition, pharmaceutically acceptable auxiliary substances, such as pH adjusting and buffering agents, osmotic pressure adjusting agents, stabilizers, wetting agents and the like, are also publicly available.

Suitable carriers include, but are not limited to, water, dextrose, glycerol, saline, ethanol, and combinations thereof. The carrier can contain additional agents, such as wetting and emulsifying agents, pH buffering agents or adjuvants, which enhance the effectiveness of the formulation. Topical vehicles include liquid petroleum, isopropyl palmitate, polyethylene glycol, ethanol (95%), polyoxyethylene monolaurate (5%) in water, or lauryl sulfate in water Sodium (5%). Other materials such as antioxidants, humectants, viscosity stabilizers, and similar agents may be added as desired.

On the other hand, the composition according to the present invention is preferably a food composition, which can be a health food or a functional food.

The compositions of the invention may be administered via any known method. For example, the pharmaceutical composition of the present invention in the form of an injection solution can be administered to an animal body.

In addition, the compositions of the present invention are preferably delivered orally, by systemic digestion and circulatory delivery to deliver the compositions according to the present invention to the desired site of action.

Other features of the present invention will be apparent from the following examples and claims. Those skilled in the art can understand that the present invention has various implementation methods, and the following examples are only for illustration but not as limitation of the present invention.

Example 1: Preparation of Water Clove Extract

Firstly, grind the cloves and immerse them in the extraction working liquid at a low temperature of 2-10°C for 4-12 hours, and then heat them at a temperature of 50-100°C for 10-60 minutes to obtain the extract and residue of cloves . The working liquid is water or ethanol solution. Then, the residue was separated by centrifugal filtration. Next, the separated residue is immersed in another extraction working liquid, and the extraction working liquid is heated at a temperature of 50-100°C for 10-60 minutes and then centrifugally filtered to produce a secondary extract of cloves monohydrate , and then dissolve the water clove secondary extract into the water clove extract. After that, use a drying process to dry the water clove extract to a water clove extract; wherein, when the extraction working liquid is water, freeze-drying can be used, and when the extraction working liquid is ethanol solution, decompression can be used Concentration method. Finally, the water clove extract is redissolved with the extraction working liquid, so that the concentration of the water clove extract is 0.005 to 0.5%.

Example 2: Water clove extract improves memory function

Drosophila learning and memory experiment (Pavlovianolfactory associative learning&memory) was used to study the effect of water clove extract on memory function, mainly using the way of Drosophila olfactory aversion learning to teach Drosophila to remember the correlation between odor and slight electric shock stimulation: in Drosophila smell An electric shock is given when a smell is detected, and then the fruit fly will run away when it smells the smell, so as to obtain a score for the correct choice, so as to judge the learning and memory function based on this test. The fruit flies were raised in an environment of 25°C, and the fruit flies were divided into four groups. The concentration of clove extract contained in the food fed to the control group was 0%, and the concentration of clove extract contained in the food fed to the experimental group was respectively 0.005%, 0.01% and 0.1%, on the 20th day, the learning and memory function experiments were carried out on Drosophila. Before each experiment, each group of fruit flies was placed in an empty tube without food. There were about 100 fruit flies in each group (mixed male and female), and they were placed in the darkroom 30 minutes before the experiment. The odors used in this experiment were 3-Octanol (3-Oct) and 4-Methylcyclohexanol (4-MCH), which were dissolved at a ratio of 1.5:1000 and 1:1000 respectively. In mineral oil, it is an odor that fruit flies hate. In order to avoid the influence of the phototaxis of Drosophila on the experimental results, the experiment was carried out in a dark room. First, flies were exposed to the first odor 3-OCT for 60 s, alternating cycles of 1.5 s shocks (at 70 volts) with 3.5 s rest followed by rest After 45 s, the flies were exposed to another odorant, 4-MCH, for 60 s, and no shock was given during the exposure to 4-MCH. After resting for 30 seconds, the fruit flies were subjected to a two-minute T-maze experiment to test their choice of two odors: the odors of 3-OCT and 4-MCH were respectively provided at both ends of the T-maze, Then let the fruit flies enter the T-shaped maze, and test the choice of the fruit flies to 3-OCT and 4-MCH, so as to judge the memory function of the fruit flies. The memory index was calculated by subtracting the number of flies choosing the taste 3-OCT from the number of flies choosing the taste 4-MCH, dividing by the total number of flies and multiplying by 100%. In the manner of the first step above, the two odor conditions (electric shock) were exchanged for another experiment, and then the results of the two experiments were averaged to obtain the average memory index. The results are shown in Figure 1, the memory index of the control group was lower than 40%, the memory index of the group whose concentration of clove extract was 0.005% was higher but lower than 60%, and the group whose concentration of clove extract was 0.01% The memory index of the group with 0.1% concentration is higher (more than 60%), and the memory index of the group with the concentration of 0.01% water clove extract is the highest.

Example 3: Water clove extract delays neurodegeneration

Experiments were carried out on 3-month-old progeria mice (senescence-accelerated mouseprone-8, SAMP8, hereinafter referred to as mice), adding water clove extract to the drinking water of the mice, and performing memory tests after 3-4 months; the experiment The mice used are short-lived and suffer from memory loss, making them suitable experimental animal models for Alzheimer's disease (neurodegeneration). The mouse memory experiment was first to place mice drinking 0% and 0.01% water clove extract individually in an open space box (20x20x10 inches) during the three-day training period to make the mice accustomed to the open space box; then, Two different training objects were placed in the open space box during the training period and the mice were allowed to explore the two different training objects for 5 minutes. When the memory test was performed after a period of time, the mice were placed in the same open space box, and one of the training objects used during the training was replaced with a new object, and the mice were allowed to explore freely for 5 minutes, and the mice were recorded. Time spent exploring training objects and new objects. The object cognition index is divided by the time the mouse explores the training object or the new object by the sum of the time exploring the training object and the new object and multiplied by 100%. As shown in Figure 2, when the memory test was carried out one hour after the end of the training period, the mice drinking 0% and 0.01% water clove extract had significantly more object cognition indexes for the new objects than the training objects The cognitive index, where the exploration time of the novel object was greater than that of the trained object, showed that both groups of mice (0% and 0.01%) could distinguish the trained object from the new object within one hour after training. When the memory test was performed two hours after the training, the mice that only drank 0.01% water clove extract could distinguish between the training object and the new object, which means that the mice remembered the training object and spent less time exploring on the training object, thus obtaining Lower object cognition index, but more curious about new objects that have not been seen, spend more time exploring new objects, and thus get a higher object cognition index. In contrast, mice that drank 0% water clove extract could not distinguish the difference between the training object and the new object, and the object cognition index of the training object and the new object was similar. The above experiments show that the extract of cloves can indeed effectively enhance the memory ability of progeria mice.

In addition, an elevated T-shaped maze was used to test the memory ability of mice. Two symmetrical ends of this T-shaped maze are open spaces (without side walls), while the other end is a closed space (with side walls on three sides around it). ). The mice were divided into an experimental group that drank clove extract containing 0.01% water, and a control group that drank a concentration of clove extract that did not contain water. Then two groups of mice were placed in the closed end of the T-shaped maze, and the mice were allowed to explore freely for 5 minutes. If the mice left the closed end within 5 minutes, the training was terminated, and the mice were placed in the T-shaped maze again after 30 seconds. For the closed end, the mice were allowed to explore freely for 5 minutes, and the training was repeated several times until the mice did not leave the closed end of the T-shaped maze for 5 minutes, so the mice learned that the closed end was a safer area than the open end. 24 hours after the training, the mice were tested for memory. As shown in Figure 3, the average time for the mice in the experimental group to stay at the closed end was about 230 seconds, while the average time for the mice in the control group to stay at the closed end was about 120 seconds . From the above results, it can be seen that since the mice in the experimental group still remember that the closed end of the T-shaped maze is safer after 24 hours, they stay at the closed end for a longer time, while the mice in the control group have poor memory, so they will not Deliberately stay at the closed end, and re-explore the T-shaped maze.

Certainly, the present invention also can have other various embodiments, without departing from the spirit and essence of the present invention, those skilled in the art can make various corresponding changes and deformations according to the present invention, but these corresponding changes All changes and modifications should belong to the protection scope of the claims of the present invention.

Claims (3)

1. A water clove extract is used to prepare a composition for enhancing memory function and delaying neurodegeneration, characterized in that the water clove extract is prepared by a method comprising the following steps: (a) extract cloves with water or ethanol at 2-10°C for 4-12 hours, then heat for 10-60 minutes at a temperature of 50-100°C to obtain cloves extract and residue; and (b) Separating the residue by centrifugal filtration, immersing it in water or ethanol, heating it at a temperature of 50-100°C for 10-60 minutes, and then centrifugally filtering it to obtain a secondary extract of cloves, and combining the water clove secondary extract and the water clove extract to obtain the water clove extract. 2. The use according to claim 1, characterized in that the composition is a pharmaceutical composition comprising an effective amount of clove extract and a pharmaceutically acceptable carrier. 3. The use according to claim 1, characterized in that, after step (b), further comprises a step (c), redissolving the water clove extract with the water or the ethanol, and the water clove extract The concentration is between 0.005% and 0.5%.