Application of chitosan in the preparation of compositions that help improve sleep

Abstract

This invention discloses the application of chitosan in the preparation of a composition that helps improve sleep, wherein the chitosan has a molecular weight of not less than 1 kDa, and the composition can improve sleep in at least three ways: increasing total sleep duration, preventing sleep fragmentation, and increasing sleep depth.

Description

Technical Field

[0001] This invention relates to the fields of medicine, food, and health food technology, and in particular to the application of chitosan in the preparation of compositions that help improve sleep. Background Technology

[0002] Sleep is an important physiological process regulated by the brain. It allows the body to rest and recover physical strength, and it also allows the brain to stop thinking activities, restore mental capacity, promote brain function development, and consolidate memory. It plays a vital role in human brain development, physical and mental health, and the maintenance of cognitive function.

[0003] Insomnia is the most prevalent sleep disorder. It is characterized by frequent and persistent difficulty falling asleep and / or maintaining sleep, leading to dissatisfaction with sleep. Insomnia can exist in isolation or be comorbid with mental disorders, physical illnesses, or substance abuse, and may be accompanied by various impairments in wakefulness. Chronic insomnia affects an individual's normal life and work, increasing the risk of various health problems. Severe sleep deprivation reduces work efficiency and alertness, and may even lead to serious accidents causing significant losses.

[0004] Currently, clinical medications for treating insomnia mainly include benzodiazepine receptor agonists (BZRAs), melatonin receptor agonists, orexin receptor antagonists, and antidepressants with hypnotic effects. While prescription drugs such as gabapentin, quetiapine, and olanzapine are effective in improving sleep, they also have significant side effects, such as dependence, cognitive impairment, tolerance, and withdrawal symptoms. Chitosan, a product of the natural polysaccharide chitin after the removal of some acetyl groups, is biodegradable and biocompatible, and is widely used in many fields, including food additives, agriculture, medicine, and drug development.

[0005] Therefore, it is of great significance to provide a new drug and health food that can help improve sleep. Summary of the Invention

[0006] The present invention aims to at least solve one of the aforementioned technical problems existing in the prior art. Therefore, the object of the present invention is to provide a composition that helps improve sleep.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] This invention provides the use of chitosan in the preparation of compositions that help improve sleep, wherein the average molecular weight of the chitosan is not less than 1 kDa.

[0009] The chitosan in this invention includes chitosan and / or oligochitosan (chitosan oligosaccharides), and the average molecular weight in this invention is exponential average molecular weight.

[0010] In some embodiments, the average molecular weight of the chitosan is not less than 3 kDa.

[0011] In some embodiments, the average molecular weight of the chitosan is not less than 30 kDa.

[0012] In some embodiments, the degree of deacetylation of the chitosan is ≥85%.

[0013] In some implementations, improving sleep refers to improving insomnia.

[0014] In some embodiments, the improvement in sleep includes at least one of the following:

[0015] Increase total sleep duration;

[0016] Prevent sleep fragmentation;

[0017] Increase sleep depth.

[0018] In some embodiments, the composition is a drug, and the drug may also contain other pharmaceutically active ingredients.

[0019] In some embodiments, the composition is a drug, and the drug may also contain a pharmaceutically acceptable carrier.

[0020] In some embodiments, the composition is a health food product, which may also contain a carrier acceptable to health foods.

[0021] In some embodiments, the carrier is selected from the group consisting of: solubilizers, binders, controlled-release polymers, disintegrants, colorants, flavorings, antioxidants, fillers, suspending agents, and / or surfactants.

[0022] In some embodiments, the drug may be formulated in a dosage form selected from the group consisting of tablets, pills, capsules, granules, powders, chewing gums, suspensions, emulsions, suppositories, or solutions.

[0023] In some embodiments, the route of administration of the drug is oral and / or parenteral.

[0024] In some embodiments, the route of administration of the drug is oral and / or parenteral.

[0025] In some embodiments, the parenteral route includes intradermal, buccal, sublingual, pulmonary, transdermal, transmucosal, intravenous, subcutaneous, intraperitoneal, and / or intramuscular injection.

[0026] In some embodiments, the health food can be made into a dosage form selected from the group consisting of: tablets, pills, capsules, granules, powders, chewing gums, suspensions, emulsions, suppositories, or solutions.

[0027] The beneficial effects of this invention are:

[0028] This invention provides the application of chitosan in the preparation of compositions that help improve sleep, wherein the active ingredient in these compositions is chitosan with a molecular weight of not less than 1 kDa. In this invention, the effect of chitosan with a molecular weight of not less than 1 kDa on improving caffeine-induced insomnia was verified using the model organism Drosophila. When the molecular weight of chitosan is not less than 1 kDa, sleep duration increases, the number of sleep episodes increases, and the duration of each sleep episode decreases, indicating that chitosan with a molecular weight of not less than 1 kDa can improve sleep in at least three ways: increasing total sleep duration, preventing sleep fragmentation, and increasing sleep depth. Attached Figure Description

[0029] Figure 1 This is a schematic diagram of a fruit fly activity monitoring system.

[0030] Figure 2 The effect of chitosan of different molecular weights on sleep duration in Drosophila. Note: Data are presented as mean ± SEM (n=16). Normal: blank group, Caffeine: caffeine model group, Caffeine+COS 1K: COS1K group (addition amount 0.375%), Caffeine+COS 3K: COS3K group (addition amount 0.375%), Caffeine+COS 30K: COS30K group (addition amount 0.375%).

[0031] Figure 3 The effect of chitosan of different molecular weights on the number of sleep cycles in Drosophila. Note: Data are presented as mean ± SEM (n=16). Normal: blank group, Caffeine: caffeine model group, Caffeine+COS 1K: COS1K group (addition amount 0.375%), Caffeine+COS 3K: COS3K group (addition amount 0.375%), Caffeine+COS 30K: COS30K group (addition amount 0.375%).

[0032] Figure 4The effect of chitosan of different molecular weights on the single sleep time of Drosophila. Note: Data are presented as mean ± SEM (n=16). Normal: blank group, Caffeine: caffeine model group, Caffeine+COS 1K: COS1K group (addition amount 0.375%), Caffeine+COS 3K: COS3K group (addition amount 0.375%), Caffeine+COS 30K: COS30K group (addition amount 0.375%). Detailed Implementation

[0033] The present invention will be further described in detail below through specific embodiments. Unless otherwise specified, the raw materials, reagents, or apparatus used in the embodiments and comparative examples are all available from conventional commercial sources or can be obtained by existing technical methods. Unless otherwise specified, the test or experimental methods are conventional methods in the art.

[0034] In this invention, the medicine and / or health food that helps improve sleep uses chitosan with a molecular weight of not less than 1 kDa as the effective active ingredient. The chitosan mentioned in this invention is chitosan and / or oligochitosan (chitosan oligosaccharides). When the composition contains an effective amount of chitosan with a molecular weight of not less than 1 kDa, it can improve sleep.

[0035] The effective amount refers to the amount that will result in improved sleep when used. There are no particular limitations on the subjects who ingest or administer the sleep-improving composition of this invention (also referred to as the subject of administration).

[0036] Chitosan, also known as chitosan polysaccharide, is a linear polysaccharide prepared from chitin through deacetylation. It possesses properties such as biodegradability, cell affinity, and biological effects. As an alkaline polysaccharide, it exerts biological effects including lowering blood lipids and blood sugar, antioxidation, anti-inflammation, antibacterial and bacteriostatic effects, and immunomodulation. Chitosan oligosaccharide (COS), also called chitosan oligosaccharide, is produced by enzymatic hydrolysis of chitosan using papain (or papain and cellulase), filtration, and spray drying. Its main component is oligoglucosamine with a degree of polymerization between 2 and 20. It is an alkaline, water-soluble oligosaccharide with various important biological activities, such as preventing and treating metabolism-related diabetes and hyperlipidemia, anti-tumor activity, alleviating degenerative diseases like Alzheimer's disease and osteoarthritis, and enhancing immunity.

[0037] The molecular weight of a polymer can affect its activity. In this application, it was unexpectedly discovered that chitosan with a molecular weight of not less than 3 kDa has a better effect on improving sleep, and chitosan with a molecular weight of not less than 30 kDa is preferred.

[0038] In this invention, improving sleep refers to improving insomnia, and more specifically, it refers to increasing total sleep duration, preventing sleep fragmentation, and / or increasing sleep depth. As shown in the examples described later, in fruit flies with caffeine-induced insomnia, a shortening of sleep duration, an increase in the number of sleep episodes, and a shortening of the duration of each sleep episode were observed. Compared to fruit flies that did not ingest chitosan, after ingesting chitosan with a molecular weight of at least 1 kDa, the fruit flies showed an increasing trend in sleep duration, a decreasing number of sleep episodes, and an increasing duration of each sleep episode. Therefore, chitosan with a molecular weight of at least 1 kDa has a sleep-improving effect, and can improve sleep quality in at least three ways: increasing total sleep duration, preventing sleep fragmentation, and increasing sleep depth. It can be used as an effective ingredient in compositions for improving sleep. Compared to fruit flies that did not ingest chitosan, the sleep duration of fruit flies significantly increased when ingesting chitosan with a molecular weight of at least 3 kDa. Compared to fruit flies that did not ingest chitosan, the sleep duration and duration of each sleep episode significantly increased when ingesting chitosan with a molecular weight of at least 30 kDa, while the number of sleep episodes significantly decreased.

[0039] The materials and equipment used in the embodiments of this invention are as follows:

[0040] The 1 kDa chitosan oligosaccharide was selected from Shandong Weikang Biotechnology Co., Ltd. (COS1K, product number HS161023), the 3 kDa chitosan oligosaccharide was selected from Shanghai Yien Chemical Technology Co., Ltd. (COS3K, product number RH330031), the 30 kDa chitosan was selected from Shanghai Maclean Biochemical Technology Co., Ltd. (COS30K, product number C15709246), agar (Biofroxx), yeast powder (Hubei Angel Yeast), wheat germ (commercially available), sucrose (commercially available), corn flour (commercially available), propionic acid (Shanghai Guoyao Group), and fruit fly activity monitor (Trikinetics, USA, model DAM5).

[0041] The number-average molecular weight of COS1K is 1490 Da, that of COS3K is 3550 Da, and that of COS30K is 34850 Da, with a degree of deacetylation of ≥85%.

[0042] Example 1: Effect of chitosan on sleep in fruit flies

[0043] Using the fruit fly (Drosophila melanogaster) to establish human disease research models is gradually becoming a new research highlight. Because fruit fly sleep exhibits significant similarities to human sleep in terms of behavioral manifestations, drug responses, and gene expression, coupled with its genetic advantages, the fruit fly has become a novel model organism for sleep research.

[0044] A 24-hour continuous monitoring of fruit fly activity was conducted using the Drosophila Activity Monitoring System (DAMS; Trikinetics Inc., USA). The DAMS system consisted of a monitor and 32 individual monitoring tubes, each 65 mm long, with food placed at one end and a breathable stopper at the other. The glass tubes were inserted into a device with an infrared emitter. As each fruit fly moved back and forth within the tube, its movement through the middle of the tube was recorded due to the interruption of the infrared beam. The fruit fly's sleep state was determined by its behavior, specifically any period of stillness lasting 5 minutes or more. The effects of chitosan on fruit fly sleep were evaluated using the number of nocturnal activities, sleep duration, number of sleep episodes, and duration of each sleep episode as key performance indicators. Monitoring was conducted for two consecutive days in a dark environment.

[0045] Laboratory animals and their rearing

[0046] Wild-type Drosophila melanogaster (Canton S) was obtained from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences. The flies were reared in an artificial climate chamber (temperature set at 25 ℃, relative humidity set at 60%), with 12 hours of daylight and 12 hours of darkness per day, in a day-night cycle.

[0047] Preparation of Drosophila test culture medium

[0048] The culture medium formula for the Drosophila experiment was 2.5 mL propionic acid, 5 g agar, 25 g sucrose, and 500 mL water. Sucrose and agar were accurately weighed into a beaker and transferred to an iron pot. Water was added, and the mixture was heated to boiling. After boiling, it was cooled, and propionic acid was added. The mixture was stirred thoroughly to obtain a blank culture medium (normal control group). Based on the blank culture medium, a caffeine-containing medium (caffeine model group) was prepared according to the specified mass ratio. Based on the caffeine culture medium, experimental culture media containing 0.375% COS1K, COS3K, and COS30K were prepared according to the specified mass ratio.

[0049] The basic culture medium formula is as follows: 40g corn flour, 25g sucrose, 20g yeast powder, 15g wheat germ, 5g agar, 2.5mL propionic acid, and 500mL water. Accurately weigh the corn flour, sucrose, yeast powder, wheat germ, and agar into a beaker, transfer them to an iron pot, add water, heat to boiling, and after boiling, cool. Then add propionic acid and stir well to obtain the basic culture medium.

[0050] Fruit fly sleep monitoring

[0051] Newborn male fruit flies within 8 hours were selected and placed in basal culture medium for rearing. After 3 days of normal culture, the flies were transferred into fruit fly activity monitoring tubes containing different intervention media, and then placed into a fruit fly activity monitoring instrument. A total of 5 groups were set up, as shown in the table below:

[0052]

[0053] Each group was equipped with one monitor, with each monitor corresponding to 32 individual monitoring tubes. One fruit fly was placed in each tube, with culture medium at one end and a vent plug at the other end. The tubes were placed in a temperature- and humidity-controlled incubator and connected to a fruit fly activity monitoring device to continuously monitor the fruit fly's activity and sleep status for two days. The fruit fly's sleep status can be defined by its activity behavior; any period of stillness lasting 5 minutes or more is considered as the fruit fly being in a sleep state.

[0054] Each fruit fly activity monitor can simultaneously monitor the activity of 32 fruit flies. The monitor has a total of four infrared beams used to monitor fly activity (e.g., ...). Figure 1 As shown, the fruit fly activity monitor emits an infrared beam from one side and receives it from the other. When a fruit fly is active inside the tube, it blocks the reception of the infrared beam. The status of the blocked beam is transmitted back to the computer once per minute to monitor the fruit fly's activity. Monitoring data is transmitted to the corresponding computer in real time. The fruit fly's sleep state is determined by its behavior, specifically any period of stillness lasting 5 minutes or more. Monitoring was conducted for two consecutive days using the number of nocturnal activities, sleep duration, number of sleep episodes, and duration of each sleep episode as evaluation indicators. All monitoring was carried out in a dark environment.

[0055] Data processing and analysis

[0056] For the Drosophila sleep study, GraphPad Prism 9.0 software was used to generate images, and SPSS 20.0 software was used for statistical analysis of the data. The Duncan test in one-way ANOVA was employed. Data are expressed as mean ± SEM. p <0.05 indicates a significant difference between groups, with different letters representing significant differences between different groups.

[0057] Test results

[0058] Effects of different molecular weights of chitosan oligosaccharides and chitosan on the sleep duration of fruit flies

[0059] Regarding sleep duration ( Figure 2 On day 1, compared with the normal group of fruit flies, the caffeine group of fruit flies had significantly reduced nighttime sleep duration. p<0.05 indicates that the caffeine-induced insomnia fruit fly model was successfully established; compared with the caffeine group, the nighttime sleep duration of fruit flies in the COS1K, COS3K, and COS30K groups was significantly increased ( p <0.05); compared with the COS1K and COS3K groups, the nighttime sleep duration of fruit flies in the COS30K group was significantly increased ( p <0.05). On day 2, compared with the normal group of fruit flies, the caffeine group of fruit flies had significantly reduced nighttime sleep duration ( p <0.05). Compared with the caffeine group, the nighttime sleep duration of fruit flies in the COS1K, COS3K, and COS30K groups was increased to some extent, and the nighttime sleep duration of fruit flies in the COS3K and COS30K groups was significantly increased ( p <0.05). Furthermore, with increasing chitosan molecular weight, the sleep duration of fruit flies increased, and compared to the COS1K group, the nighttime sleep duration of fruit flies in the COS30K group was significantly increased ( p <0.05). These results indicate that, within the intervention period, chitosan with a molecular weight within the tested range can prolong the sleep duration of insomnia-stricken fruit flies, and the larger the molecular weight, the better the effect on increasing the sleep duration of fruit flies.

[0060] Effects of different molecular weight chitosans on the number of sleep cycles in fruit flies

[0061] Regarding the number of sleeps ( Figure 3 On day 1, compared with the normal group of fruit flies, the number of sleep episodes in the caffeine group was significantly increased. p <0.05 indicates that caffeine induces fragmented sleep in fruit flies, leading to decreased sleep quality; compared with the caffeine group, there was no significant change in the number of sleep episodes in the COS1K and COS3K groups, while the number of sleep episodes in the COS30K group was significantly reduced. p <0.05). Compared with the COS1K and COS3K groups, the number of sleep cycles in the COS30K group of fruit flies was significantly reduced ( p <0.05). On day 2, compared with the caffeine group, the number of sleep events in the COS1K, COS3K, and COS30K groups showed a decreasing trend, and the intervention with COS30K significantly reduced the number of sleep events in the fruit flies ( p <0.05). Compared with the COS1K and COS3K groups, the number of sleep cycles in the COS30K group of fruit flies was significantly reduced ( p <0.05). These results indicate that, within the intervention period, chitosans with molecular weights within the tested range can reduce the number of sleep episodes in insomnia-prone fruit flies to some extent. Among these, COS30K showed the best effect in reducing the number of sleep episodes and the most significant improvement in sleep fragmentation.

[0062] Effects of different molecular weight chitosans on the single sleep time of fruit flies

[0063] For the duration of a single sleep episode ( Figure 4 On day 1, compared with the normal group of fruit flies, the single sleep time of the caffeine group of fruit flies was significantly reduced. p <0.05). Compared with the caffeine group, the single sleep time of fruit flies in the COS1K, COS3K, and COS30K groups all showed an increasing trend, and the single sleep time of fruit flies in the COS30K group was significantly increased ( p <0.05). Compared with the COS1K and COS3K groups, the single sleep time of fruit flies in the COS30K group was significantly increased ( p <0.05). On day 2, compared with the normal group of fruit flies, the single sleep time of the caffeine group of fruit flies was significantly reduced ( p <0.05). Compared with the caffeine group, the single sleep duration of fruit flies in the COS1K, COS3K, and COS30K groups showed an increasing trend, and the single sleep duration of fruit flies in the COS3K and COS30K groups was significantly increased ( p <0.05). Compared with the COS1K and COS3K groups, the single sleep time of fruit flies in the COS30K group was significantly increased ( p <0.05). These results indicate that, within the intervention period, chitosan with a molecular weight within the experimental range can increase the single sleep time of insomnia-sleeping fruit flies to a certain extent, and the larger the molecular weight, the more significant the increase in the single sleep time of fruit flies, and the better the effect on increasing the sleep depth of fruit flies.

[0064] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The use of chitosan in the preparation of compositions that help improve sleep, wherein the average molecular weight of the chitosan is not less than 30 kDa; The improvement of sleep mentioned above refers to the improvement of insomnia.

2. The application according to claim 1, characterized in that, The degree of deacetylation of the chitosan is ≥85%.

3. The application according to claim 1, characterized in that, The improvement in sleep includes at least one of the following: Increase total sleep duration; Prevent sleep fragmentation; Increase sleep depth.

4. The application according to claim 1, characterized in that, The composition is a drug, and the drug may also contain other active pharmaceutical ingredients.

5. The application according to claim 4, characterized in that, The composition is a drug, and the drug may also contain a pharmaceutically acceptable carrier.

6. The application according to claim 1, characterized in that, The composition is a health food product, and the health food product may also contain a carrier acceptable to health food products.

7. The application according to claim 5 or 6, characterized in that, The carrier is selected from the group consisting of: solubilizers, binders, controlled-release polymers, disintegrants, colorants, flavorings, antioxidants, fillers, and / or suspending agents.

8. The application according to claim 4, characterized in that, The drug can be formulated in dosage forms selected from the group consisting of: tablets, pills, capsules, granules, powders, chewable gums, suspensions, emulsions, suppositories, or solutions.

9. The application according to claim 4, characterized in that, The drug is administered orally and / or via parenteral route.

10. The application according to claim 6, characterized in that, The dosage forms that the health food can be made are selected from the following group: tablets, pills, capsules, granules, powders, chewable gums, suspensions, emulsions, suppositories, or solutions.

Images