Medicine for improving sleep deprivation induced dysmnesia or depressive symptoms

Abstract

The invention relates to the technical field of medical preparations, in particular to a medicine for improving sleep deprivation induced dysmnesia or depressive symptoms. It is found that the cannabinoid composition composed of cannabidiol and cannabinocene can significantly improve sleep deprivation induced dysmnesia and depression-like changes through a synergistic effect, significantly reduce microglial cell M1 subtype markers, reduce polarization of pro-inflammatory subtypes and increase polarization of anti-inflammatory M2 subtypes. According to the application, the increase of sleep deprivation induced proinflammatory cytokines can be reduced, the concentration of anti-inflammatory cytokines can be improved, and a scientific basis is provided for treating sleep deprivation induced dysmnesia and depression-like change by cannabinoid.

Description

Technical Field

[0001] The present invention relates to the technical field of medical preparations, in particular to a medicine for improving memory impairment or depression symptoms induced by sleep deprivation. Background Art

[0002] Hectic lifestyles, increased screen time, economic pressures, and aging are the main causes of sleep disorders, which are often accompanied by depressive symptoms and memory impairment. Approximately 25% of people worldwide suffer from sleep disorders, urgently requiring effective and safe treatments to alleviate depression and memory impairment. The development of active substances that improve memory function and alleviate depressive mood holds significant therapeutic potential. However, research on naturally occurring substances that improve memory and alleviate depression remains at the laboratory stage, with numerous challenges remaining regarding raw material selection, evaluation methods, pharmacokinetic mechanisms, and therapeutic efficacy.

[0003] Cannabinoids are a class of substances with broad sources, strong biological activity, and high development value. The market is flooded with a variety of cannabinoid-related products, which often claim to have effects on sleep deprivation, memory impairment, and depression. However, the actual purity, dosage, and therapeutic bioactive compounds vary greatly, and there is a lack of credible scientific evidence to support their claims of improving depressive symptoms and memory impairment caused by sleep disorders by regulating changes in neuroinflammation.

[0004] Cannabidiol (CBD), cannabigerol (CBG), and cannabichromene (CBC) are non-psychoactive cannabinoids. The human body produces small amounts of similar compounds, known as endocannabinoids. There are two main endocannabinoids: anandamide (AEA) and 2-arachidonoylglycerol (2-AG). In the body, these endocannabinoids are synthesized on demand from lipid precursors in cell membranes: N-arachidonoylphosphatidylethanolamine for AEA and diacylglycerol for 2-AG. These molecules bind to cannabinoid receptors (CB1 and CB2), which are located in the central and peripheral nervous systems.

[0005] Due to genetic factors, chronic stress, inflammation, neurological diseases, aging, improper dietary intake and lifestyle, dysregulated or impaired production of endocannabinoids can disrupt the endocannabinoid system, leading to reduced production of AEA and 2-AG. The endocannabinoid system (ECS) is involved in regulating a variety of physiological processes, including mood, appetite, pain perception, and immune response. By regulating these functions, endocannabinoids help maintain balance and stability in the body's internal environment. Studies have shown that the ECS is crucial for cognitive processes such as learning and memory, as well as mood regulation. In addition, endocannabinoids have anti-inflammatory and neuroprotective properties, making them potential targets for treating conditions such as chronic pain, neurodegenerative diseases, and anxiety disorders.

[0006] Besides the recent rediscovery of endocannabinoids by modern science, the sources of these compounds (such as hemp seeds, leaves, hemp oil, flaxseed, and chia seeds) have a deep historical roots in traditional medicine. In ancient China, cannabis was used medicinally as early as 2900 BC. The Shennong Bencao Jing, one of the earliest Chinese pharmacopoeias, recommended cannabis for a variety of ailments, including constipation, rheumatic pain, and malaria.

[0007] The ECS plays a crucial role in maintaining homeostasis, regulating many of our most vital functions. For example, CB1 receptors, primarily found in the brain, influence the release of neurotransmitters, impacting mood, memory, and pain perception. CB2 receptors, primarily located in the immune system, play a crucial role in regulating inflammation and immune responses. The ECS's ability to adapt to diverse physiological demands makes it a versatile system crucial to overall health.

[0008] Furthermore, endocannabinoids have enormous therapeutic potential. They are currently being investigated for their role in treating epilepsy, multiple sclerosis, and even cancer. Their anti-inflammatory and neuroprotective properties make them promising candidates for the development of new treatments for a variety of diseases. Summary of the Invention

[0009] This study investigated the effects of cannabidiol (CBD), cannabigerol (CBG), cannabichromene (CBC), and their combination on sleep deprivation-induced memory impairment and depression through animal experiments. The findings revealed that the combination of CBD and CBC exhibited a superior synergistic effect compared to the therapeutic effects of the three cannabinoids alone, significantly improving sleep deprivation-induced memory impairment and depression. Based on this, the following technical solution is proposed.

[0010] First, the present invention provides a composition consisting of cannabidiol and cannabichromene.

[0011] Preferably, the mass ratio of cannabidiol to cannachromene is 1:(0.8-1.2).

[0012] More preferably, the mass ratio of cannabidiol to cannachromene is 1:1.

[0013] Furthermore, the present invention provides a medicine containing the composition.

[0014] In some embodiments, the drug further includes excipients permitted in the pharmaceutical field.

[0015] In some embodiments, the excipients permitted in the pharmaceutical field include at least one of a filler, an excipient, a lubricant, a wetting agent, and a diluent.

[0016] In some embodiments, the dosage form of the drug includes tablets, capsules, granules, pills, powders, mixtures, oral drops, infusions, injections or sprays.

[0017] Preferably, the medicament is used to improve memory impairment or depression symptoms induced by sleep deprivation.

[0018] Preferably, the medicament is used to improve sleep deprivation-induced microglial M1 inflammatory polarization.

[0019] Preferably, the medicament is used to reduce the level of pro-inflammatory cytokines induced by sleep deprivation.

[0020] Preferably, the medicament is used to increase the level of anti-inflammatory cytokines induced by sleep deprivation.

[0021] Compared with the prior art, the present invention has the following beneficial effects: The present invention discovered that a cannabinoid composition consisting of cannabidiol and cannabichromene can significantly improve memory impairment and depressive-like changes induced by sleep deprivation through a synergistic effect, can significantly reduce microglial M1 subtype markers, reduce the polarization of pro-inflammatory subtypes and increase the polarization of anti-inflammatory M2 subtypes, can improve the increase of pro-inflammatory cytokines induced by sleep deprivation, and increase the concentration of anti-inflammatory cytokines. The present invention provides a scientific basis for the treatment of memory impairment and depressive-like changes induced by sleep deprivation with cannabinoids. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is the experimental result of the therapeutic effect of cannabinoids on behavioral changes induced by sleep deprivation; p<0.001 vs. control; #p<0.05, ##p<0.01, ###p<0.001 vs. SD; ^p<0.05, ^^^p<0.001vs. CBD+CBC.

[0023] Figure 2 is the experimental result of the therapeutic effect of cannabinoids on sleep deprivation-induced microglial polarization; p<0.05, p<0.001 vs. control; #p<0.05, ##p<0.01, ###p<0.001 vs. SD; ^p<0.05, ^^p<0.01, ^^^p<0.001 vs. CBD+CBC.

[0024] Figure 3 is the result of an experiment on the therapeutic effect of cannabinoids on the cytokine profile induced by sleep deprivation; p<0.01, p<0.001 vs. control; #p<0.05, ##p<0.01, ###p<0.001 vs. SD; ^p<0.05, ^^p<0.01, ^^^p<0.001 vs. CBD+CBC. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solutions and advantages of the present invention clearer, the technical solutions in the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. In the embodiments provided in this specification, those without specifying specific techniques or conditions are based on the techniques or conditions described in the literature in this area, or are based on product specifications. Reagents or instruments used are not specified in the manufacturer, and are conventional products that can be purchased through regular channels. The main reagents and instruments used in the following embodiments are as shown in Table 1.

[0026] Table 1 Main reagents and instruments

[0027] Example Sexually mature C57BL6 / J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. (SCXK 2020-001). Eighty mice were randomly divided into 10 groups of 8 mice each. Body weights did not differ significantly among the groups. Animals were treated as described in Table 2. Before the experiment, animals were acclimated to their new environment for one week. Following acclimation, they were dosed daily for 7 weeks as follows. For the first 5 weeks, animals were dosed in their normal cages. During the final 15 days, animals in each model group underwent chronic sleep deprivation (SD) and were dosed daily. Following the sleep deprivation protocol, animals underwent behavioral testing (including novel object recognition, open field, sucrose preference, and tail suspension tests), and brain tissue samples were collected for ELISA analysis. A CB1 antagonist (Rimonabant) and cannabinoids were dissolved in DMSO and then dissolved in corn oil to a final DMSO concentration of 5%. They were administered orally daily for the duration of the experiment.

[0028] Table 2 Experimental animal groups

[0029] The new object recognition test method is as follows: The effects of sleep deprivation on working memory were assessed using the novel object recognition (NOR) test. During the training phase, animals were placed in an open field for 5 minutes, and two similar objects were placed in two opposite quadrants of the field. During the testing phase, animals were placed in an open field for 5 minutes, but one of the objects used in the training phase was replaced by a novel object. After each test, the objects were wiped with 70% ethanol to avoid olfactory cues. Exploration of an object was defined as sniffing, touching, or licking the object, with the nose within 1 cm of the object. The number of explorations was recorded by video, and the discrimination index was calculated.

[0030] The open field (OF) test method is as follows: The open field test assesses depressive-like symptoms by evaluating spontaneous locomotor activity and exploratory behavior. Animals were tested in a polycarbonate open field (40 cm high, white background). Mice were gently placed in the center of the field, facing the wall, and recorded for 5 minutes. Distance traveled, number of stances, number of entries into the central area, and time spent in the central area were recorded. After each test, the field was wiped with 70% ethanol to eliminate olfactory cues.

[0031] The sucrose preference (SP) test method is as follows: A modified sucrose preference test was used to assess anhedonia in mice. After a 24-hour fast, mice were placed in a bottle containing a 1% (w / v) sucrose solution and a water solution. Consumption was recorded 24 hours later. The sucrose preference index was calculated as follows: Sucrose preference (%) = sucrose solution consumption / (sucrose solution consumption + pure water consumption) * 100%.

[0032] The tail suspension test (TS) method is as follows: Mice were suspended by their tails at a height of approximately 60 cm for 5 minutes. The time the animal ceased all active movement and remained suspended by its tail was considered an indicator of despair. The time spent immobile during the 5-minute period was recorded (in seconds) and analyzed.

[0033] The ELISA test method is as follows: The hippocampus was weighed and phosphate-buffered saline (PBS) was added at a ratio of 1:8. After tissue homogenization, the cells were centrifuged at 8000 rpm for 15 minutes at 4°C. The supernatant was collected and analyzed by ELISA according to the manufacturer's instructions.

[0034] Data analysis and graphing were performed using GraphPad Prism 10.3.0. Differences between groups were compared using one-way analysis of variance followed by Tukey's post hoc test. Results are presented as mean ± standard error of the mean (SEM).

[0035] The test results are as follows: 1. The therapeutic effects of cannabinoids on behavioral changes induced by sleep deprivation The results of behavioral tests are as follows Figure 1As shown, sleep deprivation model animals showed significant memory impairment (43.07%) compared to the control group (p < 0.001). Cannabinoid treatment significantly improved sleep deprivation-induced memory impairment compared to the sleep deprivation group, with increases in each group compared to the SD group (CBD, 37.31%; CBG, 36.53%; CBC, 33.40%; CBD + CBG, 33.32%; CBD + CBC, 39.40%; CBG + CBC, 33.29%) (p < 0.001). Among the cannabinoid-treated groups, the CBD + CBC combination was the most effective, with decreases in each group compared to the CBD + CBC group (CBD, 16.71%; CBG, 17.26%; CBC, 19.66%; CBD + CBG, 9.11%; CBG + CBC, 9.16%).

[0036] Sleep deprivation significantly induced depression-like behavioral impairments, as the number of entries into the central area was significantly reduced in sleep-deprived animals (64.08%) compared to the control group (p<0.001). Cannabinoid treatment significantly increased the number of entries into the central area compared to the sleep-deprived group (CBD, 56.86%; CBG, 43.13%; CBC, 60.78%; CBD+CBG, 43.95%; CBD+CBC, 56.03%; CBG+CBC, 45.16%). Among the cannabinoid treatment groups, the CBD+CBC combination was the most effective (CBD, 31.03%; CBG, 37.06%; CBC, 29.31%; CBD+CBG, 21.55%; CBG+CBC, 19.82%).

[0037] Analysis of the OF test results showed that sleep-deprived animals spent significantly less time in the central zone (52.59%) compared to the control group (p<0.001). Cannabinoid treatment significantly increased the number of entries into the central zone compared to the sleep-deprived group (CBD, 34.64%; CBG, 27.19%; CBC, 48.24%; CBD + CBG, 17.39%; CBD + CBC, 36.84%; CBG + CBC, 30.90%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 14.95%; CBG, 19.66%; CBC, 6.37%; CBD + CBG, 23.54%; CBG + CBC, 8.58%).

[0038] Analysis of the SP test results showed that the sleep-deprived model animals had a significantly reduced sucrose preference index (49.93%) compared to the control group (p<0.001). Cannabinoid treatment significantly increased the sucrose preference index compared to the sleep-deprived group (CBD, 28.15%; CBG, 20.34%; CBC, 41.93%; CBD+CBG, 36.20%; CBD+CBC, 44.53%; CBG+CBC, 37.26%). Among the cannabinoid treatment groups, the CBD+CBC combination was the most effective (CBD, 28.92%; CBG, 33.25%; CBC, 21.27%; CBD+CBG, 13.03%; CBG+CBC, 11.58%).

[0039] Analysis of the TS test results showed that sleep-deprived animals significantly increased their immobility time (147.41%) compared to the control group (p<0.001). Cannabinoid treatment significantly reduced immobility time compared to the sleep-deprived group (CBD, 16.15%; CBG, 12.23%; CBC, 20.92%; CBD + CBG, 23.60%; CBD + CBC, 34.71%; CBG + CBC, 23.10%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 12.95%; CBG, 18.23%; CBC, 6.51%; CBD + CBG, 8.98%; CBG + CBC, 9.42%).

[0040] These results suggest that cannabinoid treatment significantly improved sleep deprivation-induced memory impairment and depression-like behaviors, with the CBD + CBC combination being the most effective.

[0041] 2. Therapeutic effects of cannabinoids on sleep deprivation-induced microglial polarization Compared to the control group, sleep deprivation models showed a significant increase in the concentration of CD45, a marker for M1, in the hippocampus (135.73%) (p<0.001). Cannabinoid treatment significantly reduced CD45 concentrations compared to the sleep deprivation group (CBD, 34.65%; CBG, 22.32%; CBC, 33.74%; CBD + CBG, 59.61%; CBD + CBC, 88.11%; CBG + CBC, 66.77%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 22.93%; CBG, 46.11%; CBC, 24.64%; CBD + CBG, 17.85%; CBG + CBC, 12.79%).

[0042] Compared to the control group, sleep deprivation models showed a significant increase in hippocampal concentrations of the M1 marker IBA-1 (45.63%) (p<0.001). Cannabinoid treatment significantly reduced IBA-1 concentrations compared to the sleep deprivation group (CBD, 22.27%; CBG, 19.63%; CBC, 23.94%; CBD + CBG, 32.84%; CBD + CBC, 38.78%; CBG + CBC, 35.18%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 7.86%; CBG, 11.54%; CBC, 5.55%; CBD + CBG, 4.47%; CBG + CBC, 2.26%).

[0043] Compared to the control group, sleep deprivation significantly reduced ARG-1 levels in the hippocampus of sleep-deprived animals (25.52%) (p<0.001). Cannabinoid treatment significantly increased ARG-1 levels compared to the sleep deprivation group (CBD, 15.37%; CBG, 11.33%; CBC, 21.79%; CBD + CBG, 21.74%; CBD + CBC, 29.15%; CBG + CBC, 22.62%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 18.26%; CBG, 21.12%; CBC, 13.71%; CBD + CBG, 9.47%; CBG + CBC, 8.44%).

[0044] These results indicate that cannabinoid treatment significantly reduces microglial M1 marker concentrations, decreases pro-inflammatory polarization and increases polarization of the anti-inflammatory M2 subtype, with the CBD+CBC combination having the best effect.

[0045] 3. The therapeutic effects of cannabinoids on sleep deprivation-induced cytokines Compared to the control group, sleep deprivation models showed a significant increase in IL-1β concentrations in the hippocampus (209.14%) (p<0.001). Cannabinoid treatment significantly reduced IL-1β concentrations compared to the sleep deprivation group (CBD, 43.29%; CBG, 41.14%; CBC, 46.80%; CBD + CBG, 82.64%; CBD + CBC, 121.17%; CBG + CBC, 100.37%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 25.42%; CBG, 30.17%; CBC, 17.66%; CBD + CBG, 21.09%; CBG + CBC, 10.38%).

[0046] Compared to the control group, sleep deprivation models showed a significant increase in IL-6 concentrations in the hippocampus (39.93%) (p<0.001). Cannabinoid treatment significantly reduced IL-6 concentrations compared to the sleep deprivation group (CBD, 6.07%; CBG, 3.91%; CBC, 4.84%; CBD + CBG, 8.91%; CBD + CBC, 13.93%; CBG + CBC, 8.17%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 7.01%; CBG, 9.47%; CBC, 8.41%; CBD + CBG, 4.60%; CBG + CBC, 5.31%).

[0047] Compared to the control group, sleep deprivation significantly increased TNF-α concentrations in the hippocampus of the animals (91.89%) (p<0.001). Cannabinoid treatment significantly reduced TNF-α concentrations compared to the sleep deprivation group (CBD, 11.36%; CBG, 8.95%; CBC, 17.15%; CBD + CBG, 35.45%; CBD + CBC, 49.71%; CBG + CBC, 44.25%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 32.69%; CBG, 36.31%; CBC, 24.03%; CBD + CBG, 10.52%; CBG + CBC, 3.78%).

[0048] Compared to the control group, sleep deprivation significantly reduced IL-10 concentrations in the hippocampus of the animals (73.36%) (p<0.001). Cannabinoid treatment significantly increased IL-10 concentrations compared to the sleep deprivation group (CBD, 135.88%; CBG, 133.12%; CBC, 152.29%; CBD + CBG, 63.33%; CBD + CBC, 68.75%; CBG + CBC, 66.36%). Among the cannabinoid treatment groups, the CBD + CBC combination was the most effective (CBD, 26.29%; CBG, 27.15%; CBC, 21.16%; CBD + CBG, 14.79%; CBG + CBC, 7.10%).

[0049] These results suggest that cannabinoid treatment ameliorates sleep deprivation-induced increases in pro-inflammatory cytokines and increases concentrations of anti-inflammatory cytokines, with the CBD+CBC combination having the best effect.

[0050] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A composition, characterized in that It is composed of cannabidiol and cannabichromene.

2. The composition according to claim 1, characterized in that The mass ratio of cannabidiol to cannabichromene is 1:(0.8~1.2).

3. A medicine, characterized in that: It contains the composition according to claim 1 or 2.

4. The medicine according to claim 3, characterized in that The medicine also includes excipients permitted in the pharmaceutical field.

5. The medicine according to claim 4, characterized in that The excipients permitted in the pharmaceutical field include at least one of fillers, excipients, lubricants, wetting agents, and diluents.

6. The medicine according to claim 3, characterized in that The dosage forms of the medicine include tablets, capsules, granules, pills, powders, mixtures, oral drops, infusions, injections or sprays.

7. The medicine according to any one of claims 3 to 6, characterized in that The medicine is used to improve memory impairment or depression symptoms induced by sleep deprivation.

8. The medicine according to any one of claims 3 to 6, characterized in that The drug is used to improve the polarization of the M1 subtype of microglia induced by sleep deprivation.

9. The medicine according to any one of claims 3 to 6, characterized in that The drug is used to reduce the level of pro-inflammatory cytokines induced by sleep deprivation.

10. The medicine according to any one of claims 3 to 6, characterized in that The drug is used to increase the level of anti-inflammatory cytokines induced by sleep deprivation.

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