Use of morin in improving sleep

Abstract

The application discloses application of morin in improving sleep. Experiments prove that morin has no direct hypnotic effect, but can significantly shorten the pentobarbital sodium-induced sleep latency, prolong the sleep time, synergistically increase the sleep incidence with a subthreshold dose of pentobarbital sodium, relieve the anxiety-like behavior and increase the spontaneous activity. Compared with the existing chemical synthesis sleep-improving drugs, morin is derived from mulberry leaves which are homology of medicine and food, has no toxic side effects and no addiction, and raw materials are easy to obtain, so that morin has double development prospects of drugs and functional foods.

Description

Technical Field

[0001] This invention relates to the application of morin in improving sleep, and belongs to the fields of medicine and food technology. Background Technology

[0002] Sleep-wake cycles perform various basic biological functions in the human body. Sleep deprivation can impair emotional processing and regulation. A full night of sleep deprivation can lower the response threshold to negative emotional stimuli and increase the intensity of the response. Sleep deprivation can also lead to overexcitation of the autonomic nervous and endocrine systems in response to threatening stimuli. After sleep deprivation, threatening stimuli elicit a stronger heart rate response. When faced with stressors containing threatening information, the hypothalamic-pituitary-adrenal (HPA) axis releases hormones such as cortisol, accompanied by corresponding emotional responses.

[0003] Currently, methods for improving sleep mainly fall into two categories: cognitive behavioral therapy and chemically synthesized drugs. Benzodiazepines are the most commonly used sleep aids, offering short-term effects, but long-term use can lead to addiction, tolerance, and withdrawal symptoms. In contrast, the active ingredients in plant or natural preparations have fewer side effects and do not suppress the central nervous system. Plant compounds that are both medicinal and edible contain various bioactive substances, showing good efficacy in improving sleep with minimal toxicity. This invention is therefore proposed. Summary of the Invention

[0004] The purpose of this invention is to provide a safe, effective, and non-centrally depressant plant-derived sleep-improving monomer compound—morulaein—and its application in the preparation of sleep-improving products.

[0005] This invention relates to Morin, a plant-derived active monomer compound. Morin is a natural flavonol compound with the chemical formula C64-320. 15 H 10 O7, systematically named 3,5,7,2′,4′-pentahydroxyflavone, has hydroxyl groups attached to the 5 and 7 positions of ring A, the 3 position of ring C, and the 2′ and 4′ positions of ring B, respectively. The B ring exhibits a catechol structure, which endows it with good antioxidant activity and metal ion chelating ability.

[0006] Morin is mainly found in the form of glycosides in plants of the Moraceae family (such as mulberry leaves and mulberry bark), plants of the Equisetaceae family, and some fruits (such as apples, onions, and oranges). The morin used in this invention can be obtained from mulberry leaves through conventional phytochemical methods such as ethanol extraction and purification with macroporous adsorption resin, or it can be purchased directly from commercially available chemical or reagent suppliers (purity ≥ 98%).

[0007] This invention discovered that morin's mechanism of action is distinctly different from existing sleep-improving drugs (such as benzodiazepines): morin does not have a direct hypnotic effect and does not cause central nervous system inhibition or anxiety behavior. Its sleep-improving mechanism is mainly manifested in: shortening the sleep latency induced by sodium pentobarbital; prolonging the sleep time of suprathreshold sodium pentobarbital; synergistically increasing the incidence of sleep with subthreshold sodium pentobarbital; and indirectly improving spontaneous activity and sleep quality by reducing anxiety and stress.

[0008] This invention first provides the application of morin in the preparation of products that improve sleep; Preferably, the product is a drug, food, or health food; Preferably, the improved sleep performance includes at least one of the following: a) Shorten sleep latency; b) Extend sleep time; c) Increases the incidence of sleep induced by subthreshold doses of hypnotics; d) Reduce anxiety or stress-related changes in spontaneous activity behavior.

[0009] Preferably, the effective dose of morin in the product is 10 mg / kg body weight / day to 150 mg / kg body weight / day, more preferably 20 mg / kg body weight / day to 80 mg / kg body weight / day.

[0010] The present invention further provides a composition for improving sleep, comprising an effective amount of morin and pharmaceutically or food-acceptable excipients; The morin is the sole active ingredient in the composition; Preferably, the dosage form of the composition is an oral formulation.

[0011] Finally, this invention provides the use of morin in the preparation of food or pharmaceuticals having any of the following functions: Reduce the excitability of the central nervous system; Synergistic effect with the hypnotic effect of sodium pentobarbital; Improve anxiety-related behaviors caused by sleep deprivation.

[0012] Compared with the prior art, the present invention has the following beneficial effects: This invention discloses for the first time a novel use of morin in improving sleep: Through animal experiments, this invention demonstrates for the first time that morin can significantly shorten sleep latency (by 21.4% to 30.95% at doses of 20-80 mg / kg) and prolong sleep duration (by 93.19% at doses of 40 mg / kg). In conjunction with subthreshold doses of sodium pentobarbital, it increases the incidence of sleep to 70%, with effects close to those of the positive control, estazolam.

[0013] With a unique mechanism of action and no central nervous system depressant side effects: morin has no direct hypnotic effect (the incidence of sleep at each dose is 0%), and can reduce anxiety and stress in mice (the number of central grid entries is reduced by 13.96%-27.93%), unlike the central nervous system depressant, addiction, and withdrawal symptoms of existing benzodiazepines.

[0014] With few toxic side effects and readily available raw materials: Mulberry pigment is derived from mulberry leaves, a plant that is both a food and a medicine. Within the effective dosage range, there are no abnormal behaviors or deaths, indicating high safety. Mulberry leaf resources are abundant, the extraction process is mature, and it is easy to carry out industrial production. Attached Figure Description

[0015] Figure 1 The effect of morin on spontaneous activity behavior in mice of different groups.

[0016] Figure 2 The effect of morin on the sleep time induced by sodium pentobarbital in mice of different groups. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with embodiments. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0018] The source of morin in the following examples: Xi'an Huageng Biotechnology Co., Ltd.

[0019] Example 1: Effects of plant-derived monomeric compounds on direct sleep in mice of different groups 1. Drugs and reagents Test substance: Morin (purity ≥97%).

[0020] Positive control: Estazolam tablets (commercially available).

[0021] Solvent: 0.5% sodium carboxymethyl cellulose (CMC-Na) solution.

[0022] 2. Laboratory animals SPF-grade male Kunming mice, 6-8 weeks old and weighing 18-25g, were purchased from the Henan Provincial Laboratory Animal Center. Animals were housed in dry, clean plastic cages, fed standard laboratory animal diets, with free access to food and water, at an ambient temperature of 22±2℃, using a 12h light / 12h dark cycle. They were used in experiments after 5 days of acclimatization.

[0023] 3. Animal grouping and administration Mice were randomly divided into 5 groups of 10 each: Blank control group (NC): Administered by gavage 10 mL / kg bw 0.5% sodium carboxymethyl cellulose solution; Positive control group (Est): 2.5 mg / kg bw estazolam tablets (dissolved in 0.5% CMC-Na) were administered by gavage. Low-dose morin group (morin-L): 20 mg / kg bw morin (dissolved in 0.5% CMC-Na) was administered by gavage. Morin-M medium-dose group: 40 mg / kg morin by gavage; High-dose morin group (morin-H): 80 mg / kg morin was administered by gavage.

[0024] 4. Experimental Methods Mice in each group were administered the drug by gavage for 30 minutes, and the occurrence of sleep was observed. The disappearance of the righting reflex for more than 1 minute was used as the criterion for sleep. The incidence of sleep in each group of mice was recorded.

[0025] 5. Experimental Results The results are shown in Table 1. The incidence of sleep was 0% in all morin dosage groups (20, 40, 80 mg / kg) and the positive control group (2.5 mg / kg estazolam), as was the blank control group. This indicates that under these experimental conditions, morin has no direct hypnotic effect, meaning it does not improve sleep by directly inducing sleep.

[0026] Table 1. Effects of morin on direct sleep in mice.

[0027] Example 2: Effects of plant-derived monomeric compounds on spontaneous activity behavior in mice of different groups 1. Experimental Methods The open field test was used to evaluate spontaneous activity and anxiety-like states in mice. The open field test chamber was a rectangular box (50 cm on each side of the base and 40 cm in height), with the bottom divided into 25 identical small squares. The experiment was conducted in a dimly lit and quiet environment.

[0028] Mice in each group were administered the drugs via gavage for 7 consecutive days according to the grouping and administration regimen described in Example 1. Thirty minutes after the last administration, the mice were gently placed into an open-field box facing one corner, allowed to move freely for 5 minutes, and the process was recorded on video. The following indicators were statistically analyzed: Central grid entry count (number of times the limbs entered the central grid area); Central grid dwell time (total time spent in the central grid area); Number of times standing upright (number of times forelimbs leave the ground or climb onto the box wall).

[0029] 2. Experimental Results Experimental results are as follows Figure 1 As shown.

[0030] Number of times the central grid is entered ( Figure 1 Figure A in the middle section shows that, compared with the NC group, the number of times mice in the Est group entered the central grid was significantly reduced. P <0.05 indicates that estazolam induces anxiety-like behavior; the number of entries into the central grid decreased by 13.96%, 27.93%, and 10.50% in the morin-L, morin-M, and morin-H groups, respectively. P <0.05 or P <0.001), with the morin-M group showing the most significant effect.

[0031] Central grid dwell time ( Figure 1 Figure B in the middle section shows that, compared with the NC group, the dwell time in the central grid of the Est group was significantly reduced. P <0.001); the morin-M group showed a 22.12% decrease ( P <0.001), the morin-H group decreased by 11.06% ( P <0.05).

[0032] Number of times upright ( Figure 1 (Figure C): Compared with the NC group, the morin-M group had 25.87% more upright postures. P <0.001), while the number of upright movements decreased in the Est group.

[0033] The above results indicate that morin can reduce anxiety-like behavior and increase spontaneous activity in mice, with the medium dose (40 mg / kg) showing the best effect. This contrasts sharply with the anxiety-inducing effects of estazolam, suggesting that the mechanism of action of morin in improving sleep differs from that of traditional benzodiazepines.

[0034] Example 3: Effect of plant-derived monomeric compounds on suprathreshold dose-induced sleep in mice by sodium pentobarbital 1. Experimental Methods Male Kunming mice were used, grouped and administered drugs as in Example 1 (n=10 per group), and were continuously administered by gavage for 7 days. Thirty minutes after the last administration, sodium pentobarbital (50 mg / kg, suprathreshold dose) was injected intraperitoneally. The disappearance of the righting reflex was used as an indicator of sleep onset, and the following parameters were recorded: Sleep latency: the time from the injection of sodium pentobarbital to the disappearance of the righting reflex; Sleep duration: The time from the disappearance of the righting reflex to its recovery.

[0035] 2. Experimental Results Experimental results are as follows Figure 2 As shown.

[0036] Sleep latency ( Figure 2 (Figure A): Compared with the NC group, the sleep latency of the morin-L, morin-M, and morin-H groups was shortened by 21.4%, 28.57%, and 30.95%, respectively. P <0.05 or P <0.001), the Est group shortened by 23.81% ( P <0.05). This indicates that morin can significantly shorten the sleep latency induced by sodium pentobarbital in a dose-dependent manner.

[0037] Sleep time ( Figure 2 Figure B in the middle section shows that compared with the NC group, the sleep time in the morin-M group was prolonged by 93.19% ( P <0.001), although the sleep time in the morin-L and morin-H groups showed a trend of increasing, it was not statistically significant; the sleep time in the Est group was significantly increased ( P <0.001).

[0038] The above results indicate that morin (especially at a dose of 40 mg / kg) can significantly prolong the sleep time induced by a suprathreshold dose of sodium pentobarbital, and has a clear sleep-improving effect.

[0039] Example 4: Effect of plant-derived monomeric compounds on subthreshold dose-induced sleep in mice by sodium pentobarbital 1. Experimental Methods To rule out the possibility that morin indirectly prolongs sleep by inhibiting sodium pentobarbital metabolic enzymes (such as mixed-function oxidases), a subthreshold dose sodium pentobarbital hypnotic experiment was conducted. Grouping and administration were the same as in Example 1, with continuous gavage for 7 days. Thirty minutes after the last administration, sodium pentobarbital (40 mg / kg, subthreshold dose, at which the incidence of sleep in normal mice is typically less than 20%) was injected intraperitoneally. The absence of the righting reflex for more than 1 minute was used as the criterion for sleep onset, and the incidence of sleep in each group was recorded.

[0040] 2. Experimental Results The results are shown in Table 2.

[0041] The incidence of sleep in the NC group mice was 10%; the incidence of sleep in the Est-positive control group was 70%. P <0.001—NC); the incidence of sleep in the morin-L group was 40% ( P <0.01—NC); the morin-M group was 60% ( P <0.001—NC); the morin-H group was 70% ( P <0.001—NC). The incidence of sleep in the morin-H group was comparable to that in the positive control group.

[0042] Table 2. Effects of morin on sleep induced by subthreshold doses of sodium pentobarbital in mice.

[0043] Note: Compared with the NC group, P<0.01, *P<0.001.

[0044] The above results indicate that morin can effectively enhance the hypnotic effect of subthreshold doses of sodium pentobarbital, significantly increase the incidence of sleep, and the effects of the medium and high dose groups are close to those of the positive control. This suggests that the sleep-improving effect of morin is not achieved by inhibiting drug-metabolizing enzymes, but rather by acting directly on the central nervous system, synergistically exerting a hypnotic effect with sodium pentobarbital.

[0045] Example 5: Preparation of an oral sleep-improving preparation containing morin Weigh out the required dosage of morin raw material, add 0.5% sodium carboxymethyl cellulose solution, and stir until completely dissolved or evenly dispersed. Based on animal dosage conversion, the concentration of morin in the preparation can be formulated to 1-15 mg / mL, so that a dosage of 10-150 mg / kg morin can be administered at 10 mL / kg body weight.

[0046] This formulation can be used directly in animal experiments, or it can be further dried or excipients added to make oral dosage forms such as tablets, capsules, and granules for use as a drug or health food to improve sleep.

[0047] Based on the results of Examples 1-4, this invention demonstrates for the first time that: Morin has no direct hypnotic effect and does not cause central nervous system depression; Morin can reduce anxiety-like behavior and increase spontaneous activity in mice; Morusin can significantly shorten the sleep latency induced by sodium pentobarbital, prolong sleep time, and increase the incidence of subthreshold dose hypnosis; The above effects showed a good dose-response relationship within the dosage range of 20-80 mg / kg, with 40 mg / kg being the optimal dosage.

[0048] Therefore, morin, as a natural plant-derived monomeric compound, can be used to prepare drugs or functional foods that improve sleep, and has the advantages of being safe, effective, non-addictive, and having readily available raw materials.

[0049] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Application of morin in the preparation of sleep-improving products.

2. The application according to claim 1, characterized in that: The product is a drug, food, or health food.

3. The application according to claim 1 or 2, characterized in that: The improved sleep performance includes at least one of the following: a) Shorten sleep latency; b) Extend sleep time; c) Increases the incidence of sleep induced by subthreshold doses of hypnotics; d) Reduce anxiety or stress-related changes in spontaneous activity behavior.

4. The application according to any one of claims 1-3, characterized in that: The effective dose of morin in the product is 10 mg / kg body weight / day to 150 mg / kg body weight / day.

5. The application according to claim 4, characterized in that: The effective dose is 20 mg / kg body weight / day to 80 mg / kg body weight / day.

6. A composition for improving sleep, comprising an effective amount of morin and pharmaceutically or food-acceptable excipients; The morin is the sole active ingredient in the composition.

7. The composition according to claim 6, characterized in that: The composition is in the form of an oral preparation.

8. Application of morin in the preparation of food or medicine with any of the following functions: Reduce the excitability of the central nervous system; Synergistic effect with the hypnotic effect of sodium pentobarbital; Improve anxiety-related behaviors caused by sleep deprivation.

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