TW201521764A - Use of nattokinase in preparation of medicament with anti-aging and improved learning and memory abilities - Google Patents

Abstract

The present invention relates to a use of a nattokinase in the preparation of a medicament with anti-aging and improved learning and memory abilities. By administrating an effective dose of the nattokinase daily, a subject possesses a lower aging score and better learning and memory abilities.

Description

A nattokinase for preparing anti-aging and improving learning Recalling the use of drugs

The present invention relates to the use of a nattokinase, and in particular to the use of a nattokinase for the preparation of a medicament for anti-aging and improving learning and memory.

As medical technology advancement and material life improvement have improved the average life expectancy of human beings, coupled with a decline in fertility rates, many countries have gradually entered an aging society, so the health issues related to aging have gradually gained attention. Aging is an effect change in many molecules, cells, and systems. The aging of molecules and cells can be attributed to intrinsic factors (referred to as gene levels) and external factors (such as nutrient deficiencies, environment or disease). The aging of the system can be manifested in the decline or external characterization of organ function. Aging is an irreversible process, and there is currently no way to effectively prevent or improve it.

In addition, aging is often accompanied by a decline in learning and memory. Learning behavior definition refers to the process of changing the adaptation behavior of animals and the new behavior patterns in the process of contact with things in the environment. Memory can be classified into sensory memory (instant memory), short-term memory, medium-term memory, and long-term memory according to the length of storage. The outside world can be formed after learning Feeling memory, this sensory memory can only last for several hours. It must be through multiple re-learning processes to consolidate memory, make sensory memory further become short-term memory, and then repeat the process of consolidating memory to form medium-term memory, even long-term memory. The decline in learning and memory ability often causes inconvenience in elderly life, and there is currently no effective way to prevent or improve the decline in learning and memory.

Nattokinase is an extracellular protein of natto (Bacillus subtilis) isolated from soy bean mash or natto. Studies have shown that nattokinase has the ability to directly decompose fibrin like fibrin, promotes the activation of fibrinogen, and is safer than the drugs used in the treatment of vascular embolism, so nattokinase It is currently used for the treatment and prevention of vascular embolism and hypertension.

However, studies on nattokinase have rarely investigated whether nattokinase can be used for anti-aging and for improving learning and memory.

In view of this, it is urgent to provide a new use of nattokinase for anti-aging and improving learning and memory ability to develop other applications of nattokinase.

Therefore, one aspect of the present invention is to provide a nattokinase for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein nattokinase is administered to a subject in a daily orally effective dose, and the subject can be compared. Low aging index and better learning and memory ability, and then the application of nattokinase to the preparation of anti-aging and improving learning and memory ability of drugs.

According to the above aspect of the invention, a nattokinase is proposed for use in the preparation of Use of anti-aging and medicines that improve learning and memory. Nattokinase is administered to a subject at a daily orally effective dose, which allows the subject to have a lower aging index and better learning and memory ability, and then apply nattokinase to prepare a drug resistant to aging and improve learning and memory. use.

According to an embodiment of the present invention, the subject may be, for example, a human, and the daily oral effective dose of the human may be, for example, 26.70 fibrin units (FU) per kilogram of body weight per day to 266.67 fibrin per kilogram of body weight per day. Decomposition unit, 26.70 fibrinolytic units per kilogram of body weight per day to 53.33 fibrinolytic units per kilogram of body weight per day, and 53.33 fibrinolytic units per kilogram of body weight per day to 266.67 fibrinolytic units per kilogram of body weight per day.

According to another embodiment of the present invention, the subject may be a rodent. And the above-mentioned rodent can be, for example, a senescence-accelerated prone mouse-8 (SAMP8), and the daily oral effective dose of the aging promoting mouse can be, for example, 8.2 fibrinolytic units per kilogram of body weight per day to every day. The kilograms of body weight 81.5 fibrinolytic units, 8.2 fibrinolytic units per kilogram of body weight per day to 16.3 fibrinolytic units per kilogram of body weight per day, and 16.3 fibrinolytic units per kilogram of body weight per day to 81.5 fibrinolytic units per kilogram of body weight per day.

The use of a nattokinase of the present invention for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein nattokinase is administered to a subject at a daily orally effective dose, which allows the subject to have a lower aging index and Better learning and memory ability, and then the application of nattokinase to the preparation of anti-aging and improving learning and memory ability of drugs.

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The above and other objects, features, advantages and embodiments of the present invention will become more <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; A bar graph of the effect of kinase on the number of escape responses to active avoidance trials.

Figure 2 is a bar graph showing the effect of administration of nattokinase on the residence time of a single passive test in SAMP8 mice according to an embodiment of the present invention.

As described above, the present invention provides a use of nattokinase for the preparation of a medicament for anti-aging and improving learning and memory ability, and administering a daily effective dose of nattokinase to a subject, so that the subject has a lower aging index and Better learning and memory skills.

The nattokinase of the present invention has been confirmed by in vivo animal experiments that it is possible to specifically reduce the aging index and improve the learning and memory ability of the test animals after oral administration of an effective dose of nattokin for a period of time. It should be noted that the term "animal experiment" as used herein refers to the use of an aging animal, such as a 6-month-old female SAMP8 mouse, after daily oral administration of an effective dose of nattokinase for 11 weeks, the SAMP8. Compared with those who did not receive nattokinase, mice did have a lower aging score and better learning and memory ability.

The aforementioned SAMP8 mice are genetically aging animal models with natural aging and intelligent gradual decline characteristics, with an average life expectancy of about 299 days (10 Month), early aging characteristics such as stopping weight gain, hair loss and poor immune system, are often used in aging related research.

The aging index referred to herein as the present invention may include a plurality of evaluation items such as behavior of SAMP8 mice, skin, eyes, and spine. Among them, the behavioral evaluation project was to observe the escape response of SAMP8 mice within 30 seconds and the escaping reaction when pinching the back skin of the neck. The external evaluation item is to observe the condition of the hair's gloss, roughness, hair loss and skin ulcers. The eye evaluation program is to observe the edema of mucositis or eyelids around the eyes. The evaluation of the spine is to use the inspection and touch methods to determine whether the spine is bent. The scores were given according to each evaluation item to reflect the degree of aging of SAMP8 mice.

In addition, the current experimental model often evaluates the learning and memory ability of animals in the form of conditioned reflex. The principle of conditioned reflex is that when an animal is subjected to a noxious stimulus (such as an electric shock), an escape response is immediately generated, and the escape response is passive. However, if a conditioned stimulus (CS) (such as sound or light) is provided prior to noxious stimulation, an active avoidance response can be gradually formed. The learning and memory ability referred to herein as the present invention is evaluated by a single-trial passive avoidance test and an active shuttle avoidance test.

The following examples are provided to illustrate the application of the present invention, and are not intended to limit the present invention, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention.

Example 1: Establishment of animal evaluation model

In this experiment, 40 female 6-month-old female SAMP8 mice were used to establish an animal evaluation model. The feeding conditions of SAMP8 mice were carried out in accordance with the relevant experimental animal management guidelines announced by the National Institutes of Health. The SAMP8 mice were housed in a transparent plastic cage (30 (W) × 20 (D) × 10 (H) cm ³ ), and the temperature in the animal room was maintained at 25 ± 2 ° C, and the relative humidity was 65 ± 5%. In the dust-free automatic control room, the lighting cycle is controlled by an automatic timer, the dark period is from 07:00 to 19:00, and the light period is from 19:00 to 07:00. Both feed and water are fed freely.

Then, according to the "Health Food Health Function Assessment Method" of the Department of Health, 6-month-old female SAMP8 mice were randomly selected and divided into control group and low, medium and high dose experimental groups, 10 mice in each group, a total of 40 mice. The experiment lasted for 12 weeks.

The dosage of nattokinase used in the present invention is based on the recommended daily intake per kilogram of body weight of humans, and is converted into the recommended daily intake per kilogram of body weight of SAMP8 mice by means of orogastric intubation. Different doses of nattokinase were administered to SAMP8 mice to evaluate the anti-aging effect of nattokinase and improve learning and memory ability.

Taking 60 kg of adults as an example, the recommended intake of nattokinase is 3200FU per day, which is converted to a daily intake of 53FU/kg BW/day per kilogram of body weight. Next, the recommended daily intake per kilogram of body weight of humans is converted to the recommended daily intake per kilogram of body weight of SAMP8 mice based on the calculation of the body surface area of humans and mice. Among them, the intake of SAMP8 mice in the middle dose group is equivalent to one dose of the recommended intake of the human body, and its use The dose in SAMP8 mice was 16.3 FU/kg BW/day. The intake of SAMP8 mice in the low-dose group and the high-dose group was equivalent to 0.5 times and 5 times of the recommended intake of the human body, respectively, and the doses used in SAMP8 mice were 8.2FU/kg BW/day, respectively. 81.5FU/kg BW/day. The control group of SAMP8 mice did not administer nattokinase. The experimental group and sample administration methods are shown in Table 1.

The daily intake and drinking capacity of SAMP8 mice were recorded during the experiment. The locomotion test was performed at the 11th week of the experiment and the aging index of SAMP8 mice was evaluated. The single passive avoidance test and the active avoidance test were used to evaluate the SAMP8 mice. Learning memory.

The data obtained in this study were statistically analyzed using the SAS statistical software package. The values of the experimental results were expressed as mean ± mean standard error (mean ± SD). The experimental data were analyzed by one-way analysis of variance (one-way ANOVA) to determine the differences between groups, and the differences between groups were compared by Duncan multiple full-range assay. When comparing the differences between groups, the control group is used as the comparison benchmark, and the English letter a is indicated above the data of the control group. If there is no significant difference between the experimental results of the experimental group and the comparison group (p>0.05), the English letter a is indicated above the experimental group data. Experimental group When there is a significant difference (p<0.05) between the experimental results of the control group, the English letter b is indicated above the experimental group data. In order to improve the clarity, the data presented below is represented by a significant difference between the experimental group and the control group data with different English letters.

Example 2: Evaluation of the effect of nattokinase on the aging index of SAMP8 mice

The evaluation of the aging index was carried out in accordance with the method established by Takeda et al. in 1981. The aging index assessment included changes in the behavior, appearance, health of the eyes, and curvature of the spine of SAMP8 mice. Among them, the behavioral evaluation item was to observe the escape reaction in the SAMP8 mice within 30 seconds and the escape response when the skin of the neck was pinched. The external evaluation item is to observe the glossiness, roughness, hair loss and skin ulcer condition of the hair. The assessment of the health of the eye is to observe the edema of mucositis or eyelids around the eyes. Changes in spinal curvature are assessed using inspection and touch. The scores were given according to each evaluation item to reflect the degree of aging of SAMP8 mice. Each item has 5 levels, 0, 1, 2, 3, and 4, which are scored according to the definition of each level. The lower the score, the better the anti-aging effect. The aging index of different doses of nattokinase administered to female SAMP8 mice at 6 months of age was as shown in Table 2.

The results showed that the female aging index of the 6-month-old female SAMP8 mice was significantly higher than the experimental group in terms of total scores of behavior, skin, hair, eyes, spine and other aging indexes after 11 weeks of different doses of nattokinase. . This result showed that administration of nattokinase significantly reduced the occurrence of aging in SAMP8 mice (p<0.05). In addition, the degree of reduction in the aging index of SAMP8 mice was more pronounced as the dose of nattokinin administered increased.

Example 3: Assessing the effect of nattokinase on the activity of SAMP8 mice

The activity test was conducted at the 11th week of the experiment. First, the SAMP8 mouse was placed in the center of a commercially available aluminum box (for example, activity monitor video path analyzer, Coulbourn instruments Model E61-21, 25 (W) × 25 (D) × 25 (H) cm ³ ), and the local portion thereof was measured. The amount of activity activity. This test operates in a low ambient light environment and the groups are crossed. The amount of activity in the plane movement time of the SAMP8 mice (locomotion; sec/5min) was recorded by the recorder every 5 minutes, and the activity test took a total of 10 minutes. The primary purpose of this test was to understand the planar walking of SAMP8 mice as a basis for assessing their activity and to eliminate SAMP8 mice with abnormal activity (eg, those with the highest number of turns in the same direction in the same or counterclockwise direction). Since the active avoidance test and the single passive avoidance test used to evaluate the learning and memory ability are all a plane moving behavior, it is necessary to use the activity test to understand the activity of the SAMP8 mouse in advance and eliminate the abnormal activity. SAMP8 mice, in order to avoid interference with subsequent learning and memory ability assessment. The amount of activity of different doses of nattokinase administered to female SAMP8 mice at 6 months of age was shown in Table 3.

The results showed that there was no significant difference in the amount of activity between the four groups of SAMP8 mice at 0-5 minutes and 6-10 minutes. In the experiment, the activity of SAMP8 mice was recorded every 5 minutes, and the activity of SAMP8 mice decreased after 5 minutes, and the amount of activity decreased with the increase of test time. This phenomenon presumes that when SAMP8 mice are in an unfamiliar environment, there will be a strong exploration behavior at the beginning, resulting in a higher activity. However, as test time increased, SAMP8 mice gradually adapted to the new environment, and their exploration behavior decreased and the amount of activity decreased. It is speculated from the experimental results that SAMP8 mice have higher learning and memory ability behaviors 5 minutes before the experiment, so it is better to set the learning and memory ability evaluation time within 5 minutes.

Example 4: Evaluation of the effect of nattokinase on learning and memory ability of SAMP8 mice

In this experiment, 11-month-old female SAMP8 mice were administered with different doses of nattokinase for 11 weeks. The effects of nattokinase on learning and memory ability of SAMP8 mice were evaluated by active avoidance test and single passive avoidance test.

Active avoidance test

Use a commercially available aluminum box (for example, shuttle cage, Model E10-15, 35 (W) × 17 (D) × 20 (H) cm ³ , Coulbourn instruments). This aluminum box has two chambers, two chambers. The middle is separated by a small gate (7.5 (W) × 6.5 (D) cm ² ) and can be connected. The bottom of the box is provided with metal rods arranged in parallel at intervals of 1 cm, and an electric current device is connected. The experimental process is controlled by the computer to control the time, sound and light and electric shock, and finally output the result. During the test, the SAMP8 mice were first placed on one side (no light appeared in the chambers on both sides), and after 10 seconds of adaptation (this is the intertrial interval), the computer detects the SAMP8 mice staying. The position of the box is located on the left or right side and gives the side 10 seconds of light and sound stimulation as a conditioned stimulus (CS). If the SAMP8 mouse is under CS and remains on the same side without any reaction, the computer gives an electric shock of 0.3 milliamperes (mA) for 5 seconds as an unconditioned stimulus (UCS). If the SAMP8 mouse enters the other side after CS, the computer immediately stops the stimulation of light and sound and does not give an electric shock. If the SAMP8 mice stay on the same side throughout the process, it means that they have not yet learned to give a shock (UCS), which is the process of learning. If a SAMP8 mouse moves to the other side under a shock (UCS), it means that it has learned not to give an electric shock, which is a manifestation of memory. The computer will automatically display the status of CS and UCS according to the SAMP8 mouse reaction. Each SAMP8 mouse will receive 5 cycles of CS and UCS as a cycle. After 15-20 minutes, the same process will be used for 4 cycles. Days, that is, 20 sound and light stimulation trainings were taken every day to observe changes in learning and memory ability of SAMP8 mice. The computer recorded the number of times SAMP8 mice were shocked and not shocked. The behavior of SAMP8 mice that are moved to the other side after being shocked is called escape responses, also known as the number of failures. The case where the mouse is stimulated by light and sound and then moved to the other side without being shocked is called avoidance response. The number of avoidance reactions and the number of failures in the 20 sound and light stimulation trainings were recorded. The fewer the number of escape reactions, the better the learning ability, and the fewer the number of escape responses, the more the number of successful avoidance reactions. The test results are shown in Figure 1.

Please refer to FIG. 1 , which is a bar graph showing the effect of nattokinase administered by SAMP8 mice on the number of escape responses in an active avoidance test according to an embodiment of the present invention, wherein the vertical axis indicates the number of escape reactions (times). The horizontal axis represents the time of the experiment (days), and the long bars 101, 103, 105, and 107 respectively represent the control Group, low dose, medium dose and high dose groups. The results showed that since the SAMP8 mice were still in the learning phase on the first day of the experiment, there was no significant difference in the number of escape responses between the groups. On the third day of the experiment, the number of escape reactions in the strip 105 (middle dose group) and the strip 107 (high dose group) was less than that in the strip 101 (control group), and significant differences were achieved. Level (p<0.05). On the fourth day of the experiment, the active learning ability of the strip 101 (control group) and the strip 103 (low dose group), 105 (medium dose group), and 107 (high dose group) all showed that the SAMP8 was small with the increase of the number of days. The rats gradually learned to escape successfully and the number of escape reactions gradually decreased, but there was no significant difference between the groups. According to the active success avoidance test, after nattokinase was administered to female SAMP8 mice at 6 months of age, the strip 105 (middle dose group) and the strip 107 (high dose group) were compared with the strip 101 (control group). There was a significant promotion of learning and memory ability in SAMP8 mice (p<0.05).

2. Single passive avoidance test

The single passive avoidance test uses the same aluminum box as the active avoidance test. The aluminum box has two chambers, and the two chambers are separated by a small gate (7.5 (W) × 6.5 (D) cm ² ) and can communicate with each other. The two chambers are respectively the bright room and the dark room. The computer can detect the position of the chamber where the SAMP8 mouse stays is located on the left or right side, and controls the time, small gate switch and electric shock, and finally outputs the result. During the test, the SAMP8 mice were first placed in the bright room, and after 10 seconds to adapt to the environment, the small gate was opened to connect the bright room to the dark room, so that the SAMP8 mice were free to explore. Since SAMP8 mice have darkening, they move toward the dark room. When the SAMP8 mouse enters the dark room, the small gate is quickly closed, and an electric shock of 0.5 microamperes for 0.5 seconds is given after five seconds. Three electric shocks are given continuously and each interval is 5 seconds, and the residence time in the bright room is recorded, that is, the learning is completed. Training process. Thereafter, the residence time in the bright room was measured 1 day, 2 days, 3 days, and 7 days after the completion of the learning training process, and the same operation method as above was used in the test, but no electric shock was given. The maximum period of each test time is 180 seconds. The longer the retention time in the bright room, the better the learning and memory ability of SAMP8 mice. The test results are shown in Figure 2.

Please refer to FIG. 2, which is a bar graph showing the effect of administration of nattokinase on the residence time of a single passive test in a SAMP8 mouse according to an embodiment of the present invention, wherein the vertical axis represents the residence time (seconds) and the horizontal axis. The time (days) is indicated, and the bars 201, 203, 205, and 207 represent the control group, the low dose group, the medium dose group, and the high dose group, respectively. The results showed that when the learning training process was completed (time is 0), there was no difference in the retention time of each group. One day after the completion of the learning training process, the longer stay time (control group) of the long strip 205 (middle dose group) and the long strip 207 (high dose group) was significantly increased (p < 0.05). 2 days, 3 days, and 7 days after the training, the retention time of each group decreased. This phenomenon may be due to passive avoidance giving electric shock during the training process, but 1 day and 2 days after the completion of the learning training process. , 3 days and 7 days did not give an electric shock, the SAMP8 mice one day after the completion of the learning training, the retention time in the bright room is longer, but as time increases, the memory retention capacity decreases, resulting in the bright room The residence time in the chain decreases. From the results of a single passive test, after nattokinase was administered to female SAMP8 mice at 6 months of age, the strip 205 (middle dose group) and the strip 207 (high dose group) were compared with the strip 201 (control group). ) significantly promoted SAMP8 mice The situation of learning memory ability (p<0.05).

In summary, the nattokinase of the present invention has been proven to be useful for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is SAMP8 mouse at a rate of 8.2 fibrin per kilogram of body weight per day. 81.5 fibrinolytic units per kilogram of body weight per day per day, 8.2 fibrinolytic units per kilogram of body weight per day to 16.3 fibrinolytic units per kilogram of body weight per day are preferred, and 16.3 fibrinolytic units per kilogram of body weight per day to daily 81.5 fibrinolytic units per kilogram of body weight are preferred. The human equivalent dose was converted to humans with 26.70 fibrinolytic units per kilogram of body weight per day to 266.67 fibrinolytic units per kilogram of body weight per day, with 26.70 fibrinolytic units per kilogram of body weight per day to per kilogram of body weight per day. The 53.33 fibrinolytic unit is preferred, and more preferably 266.67 fibrinolytic units per kilogram of body weight per day for 53.33 fibrinolytic units per kilogram of body weight per day.

It is worth mentioning that the body weight, food intake and water consumption of the SAMP8 mice in the experimental group were not significantly different from those in the control group before and after the experiment, indicating that nattokinase did not affect the food intake and drinking water of SAMP8 mice. Volume and weight changes.

In addition, feeding nattokinase did not cause damage to organs in SAMP8 mice. In other experiments, SAMP8 mice were sacrificed by eye socket sampling 12 weeks after the experiment (fasting for 8 hours before sacrifice), and blood and organs were taken for data analysis (not shown). Comparing the weight of organs in each part (including heart, liver, spleen, lungs and kidneys), the weight of each part of each experimental group There was no significant difference from the control group. The SAMP8 mice in each experimental group were observed by the naked eye after anatomical sacrifice. The results showed that the appearance of each organ in each experimental group was not significantly enlarged, and the color of each organ was also normal.

Moreover, administration of nattokinase did not cause any adverse effects on the blood quality of SAMP8 mice. In other experiments, the project included total protein, albumin, glucose, triglyceride, alanine transaminase, glutamic acid transaminase, total cholesterol, high-density lipoprotein and low-density lipoprotein. Blood biochemical measurements (not shown), blood biochemical values of each experimental group were not significantly different from the control group.

However, it should be added that the present invention exemplifies a specific animal model, a specific application mode, a specific material or a specific device, etc., and illustrates the use of the nattokinase of the present invention for preparing a drug resistant to aging and improving learning and memory. However, it is to be understood by those skilled in the art that the present invention is not limited thereto, and other animal models, other modes of administration, and other grades may be used in the nattokinase of the present invention without departing from the spirit and scope of the present invention. Equivalent materials or other equipment are carried out.

It can be seen from the above embodiments of the present invention that the present invention has the advantages of using the above-mentioned daily oral effective dose of nattokinase to administer the test object, thereby effectively reducing the aging index and increasing the learning and memory ability, thereby applying the anti-aging and improving learning and memory ability. The use of the drug.

While the invention has been described above in terms of several embodiments, it is not intended to limit the scope of the invention, and the invention may be practiced in various embodiments without departing from the spirit and scope of the invention. Change and retouch, so the scope of protection of the present invention is attached to the patent application The definition of the enclosure shall prevail.

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Claims (10)

A use of nattokinase for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the nattokinase is administered to a subject in a daily orally effective dose. The use of nattokinase as described in claim 1 for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the subject is a human. The use of nattokinase according to claim 2, for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 26.70 fibrinolytic units per kilogram of body weight per day ( Fibrin units; FU) to 266.67 fibrinolytic units per kilogram of body weight per day. The use of nattokinase according to claim 2, for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 26.70 fibrinolytic units per kilogram of body weight per day to 53.33 fibrinolytic units per kilogram of body weight per day. The use of nattokinase according to claim 2, for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 53.33 fibrinolytic units per kilogram of body weight per day to 266.67 fibrinolytic units per kilogram of body weight per day. The use of nattokinase according to claim 1, wherein the subject is a rodent. The use of nattokinase as described in claim 6 for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the rodent is a senescence-accelerated prone mouse-8 (SAMP8). The use of nattokinase according to claim 6 for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 8.2 fibrinolytic units per kilogram of body weight per day to 81.5 fibrinolytic units per kilogram of body weight per day. The use of nattokinase according to claim 6 for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 8.2 fibrinolytic units per kilogram of body weight per day to 16.3 fibrinolytic units per kilogram of body weight per day. The use of nattokinase according to claim 6 for the preparation of a medicament for anti-aging and improving learning and memory ability, wherein the daily oral effective dose of the subject is 16.3 fibrinolytic units per kilogram of body weight per day to 81.5 fibrinolytic units per kilogram of body weight per day.