Application of capric acid monoglyceride in preparation of product for improving cognitive impairment

Abstract

The invention discloses application of capric acid monoglyceride in preparation of a product for improving cognitive impairment. Experiments prove that the capric acid monoglyceride has the effects of enhancing the memory ability of the cognitive impairment, reducing the level of neuroinflammatory factors in the brain tissue of the cognitive impairment and improving the gene expression level of neurotrophic factors in the brain tissue of the cognitive impairment, and can be used for developing products for improving the cognitive impairment. The product is a medicine for improving cognitive impairment or a health food for enhancing memory.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical and health product technology, specifically to the application of decanoic acid monoglyceride in the preparation of products for improving cognitive impairment. Background Technology

[0002] Lipid metabolism disorders are a core mechanism of obesity-related cognitive decline. The dynamic balance of lipids in the brain is crucial for neuronal function and synaptic plasticity, while lipid overload caused by obesity leads to decreased hippocampal synaptic plasticity, elevated levels of neuroinflammatory factors (such as IL-6 and TNF-α), and impaired blood-brain barrier (BBB) ​​integrity. High-fat diets (HFD) are a major contributing factor; HFD induces peripheral insulin resistance, leading to Aβ deposition and Tau protein hyperphosphorylation in the brain, thus disrupting hippocampal synaptic plasticity. However, how HFD leads to cognitive impairment through the "metabolism-inflammation-neural damage" axis is not fully understood. Existing drugs such as insulin sensitizers (metformin) and anti-inflammatory drugs (non-steroidal anti-inflammatory drugs) can improve neuroinflammation, but their effects are limited and they have hepatotoxic and nephrotoxic effects. Poor adherence to diet control / exercise, and unstable efficacy of probiotics / prebiotics mean that current methods cannot simultaneously address the multiple pathological links of "metabolic disorder-neural inflammation-synaptic damage." Glyceryl decanoate (GMD) has been widely shown to regulate obesity-induced glucose and lipid metabolism disorders, intestinal damage, and gut microbiota regulation, but no studies have been reported on the role of GMD in improving cognitive impairment in HFD-induced obese mice. Summary of the Invention

[0003] Based on the first discovery that decanoic acid monoglyceride has a novel function in improving cognitive impairment, this application provides a new use for decanoic acid monoglyceride, which can be used to prepare drugs for improving cognitive impairment or health foods for enhancing memory.

[0004] This application provides the use of decanoic acid monoglyceride in the preparation of a medicament for improving cognitive impairment.

[0005] Optionally, decanoic acid monoglyceride is the sole active ingredient in the drug.

[0006] Optionally, the drug comprises a therapeutically effective amount of decanoic acid monoglyceride or a pharmaceutically acceptable carrier or excipient.

[0007] Therapeutic effective dose refers to the range of doses within which a drug, in clinical application, produces a clear therapeutic effect on the target disease while adverse reactions remain within an acceptable range. This range can vary depending on the severity of the disease and the physical condition, age, weight, and sex of the subject to be treated. Pharmaceutically acceptable carriers or excipients refer to all substances in a pharmaceutical preparation, other than the active ingredient, that have undergone reasonable and safe evaluation. These substances themselves typically do not have a therapeutic effect.

[0008] When used in pharmaceutical preparations, the decanoic acid monoglyceride can be prepared with pharmaceutically acceptable excipients using conventional pharmaceutical formulation methods. Pharmaceutically acceptable excipients include buffer solutions, emulsifiers, suspending agents, disintegrants, disintegrants, dispersants, binders, excipients, stabilizers, chelating agents, diluents, gelling agents, preservatives, wetting agents, lubricants, absorption delay agents, liposomes, etc., which can be selected according to different needs.

[0009] Optionally, the dosage form of the drug is a tablet, granule, oral liquid preparation, injection preparation, or capsule preparation.

[0010] Optionally, the improvement of cognitive impairment includes at least one of the following: (1) Enhance the memory ability of people with cognitive impairment; (2) Reduce the level of neuroinflammatory factors in the brain tissue of individuals with cognitive impairment; (3) Increase the expression levels of neurotrophic factors and cognitive-related genes in the brain tissue of individuals with cognitive impairment.

[0011] Optionally, the neuroinflammatory factors include at least one of TNF-α (tumor necrosis factor-α), IL-1β (interleukin-1β), IL-6 (interleukin-6), MCP-1 (monocyte chemoattractant protein-1), and iNOS (inducible nitric oxide synthase).

[0012] Optionally, the neurotrophic factor includes at least one of BDNF (brain-derived neurotrophic factor), NT-3 (neurotrophic factor-3), and NT-4 (neurotrophic factor-4), all of which are secreted neurotrophic factors of the NGF family.

[0013] Optionally, the cognitive-related genes include at least one of the following: PSD-95 (synaptic scaffold protein) gene, Fxr1 (RNA-binding protein) gene, and Fxr2 (RNA-binding protein) gene.

[0014] Optionally, the cognitive impairment includes at least one of the following behavioral impairments: memory impairment, visuospatial impairment, and computational impairment.

[0015] Optionally, the cognitive impairment may be obesity-related cognitive decline.

[0016] Optionally, the cognitive impairment may be Alzheimer's disease.

[0017] This application also provides the use of decanoic acid monoglyceride in the preparation of health food products that enhance memory.

[0018] Optionally, the health food includes decanoic acid monoglyceride or a food-grade acceptable carrier or excipient.

[0019] When used in the preparation of health food products, the decanoic acid monoglyceride can be prepared with food-acceptable excipients according to conventional food preparation methods. These excipients include solvents, binders, fillers, sugars, and sweeteners, which can be selected according to different needs. The health food product can be any of the following types: tablets, capsules, oral liquids, compressed candies, solid beverages, pills, granules, decoctions, pastes, syrups, drop pills, or tea bags.

[0020] This application has the following beneficial effects: This application demonstrates that decanoic acid monoglyceride can increase motor and spatial exploration abilities in HFD-induced mice, improve anxiety-like behavior in HFD-induced mice, enhance cognitive and memory abilities, and reduce the gene expression levels of neuroinflammatory factors and increase neurotrophic factors, thereby improving cognitive impairment in HFD-induced mice.

[0021] This application is the first to discover that decanoic acid monoglyceride, a medium-chain fatty acid monoglyceride, has the effect of improving cognitive impairment, and preliminarily reveals its potential mechanisms of improving cognitive impairment through improving motor spatial exploration ability, anxiety-like behavior, enhancing cognitive ability, and improving neuroinflammation. Attached Figure Description

[0022] Figure 1 The figure shows the effect of decanoic acid monoglyceride on HFD-induced motor and spatial exploration abilities in mice (a: representative movement trajectory in the open field; b: total activity distance in the open field; c: activity distance in the center of the open field; d: dwell time in the central region; e. percentage of time spent in the central region compared to the surrounding region).

[0023] Figure 2 The figure shows the effect of decanoic acid monoglyceride on cognitive and memory abilities in HFD-induced mice (a: representative trajectory for new object recognition; b: new object recognition discrimination index; c: new object recognition preference index).

[0024] Figure 3 Figure 1 shows the effect of decanoic acid monoglyceride on HFD-induced anxiety-depression-like behavior in mice (a: representative movement trajectory in the elevated cross maze; b: percentage of open-arm movement distance; c: ratio of open-arm movement distance to closed-arm movement distance; d: percentage of open-arm dwell time; e: ratio of open-arm movement time to closed-arm movement time).

[0025] Figure 4 The figure shows the effect of decanoic acid monoglyceride on HFD-induced neuroinflammation in mice (af: mRNA expression levels of TNF-α, IL-1β, IL-6, COX-2, MCP-1, and iNOS).

[0026] Figure 5 shows the effect of decanoic acid monoglyceride on HFD-induced striated plasticity in mouse brain (af: mRNA expression levels of BDNF, PSD-95, NT-3, NT-4, Fxr1, and Fxr2). Detailed Implementation

[0027] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0029] The glyceryl monodecanoate (GMD) used in the following examples is commercially available and is CAS number 26402-22-2.

[0030] Example 1 Evaluation of the functional activity of decanoic acid monoglyceride in improving cognitive impairment: Functional evaluation was conducted using mice as animal models.

[0031] Animal feeding and grouping: Thirty-six healthy male C57BL / 6 mice aged 6-8 weeks were fed a normal diet for 7 days to acclimatize to the experimental environment (12h day / night cycle). After the acclimatization period, they were randomly divided into three groups: a normal group (NCD, fed a normal diet), a model group (HFD, fed a 60% kcal high-fat diet), and an intervention group (GMD, fed a 60% kcal high-fat diet supplemented with 1600 mg / kg GMD). Each group contained 12 mice, divided into 3 cages of 4 mice each. The environment was (22 ± 2 °C) with humidity ranging from 30% to 70%. Mice had free access to food and water. The experiment lasted for 14 weeks, and body weight and food intake were recorded regularly. Behavioral experiments were conducted in week 8. Mice were dissected in week 9, and mouse brain tissue was cryopreserved at -80°C. All methods used in this animal experiment were reviewed and approved by the Ethics Committee of Zhejiang University of Traditional Chinese Medicine (20250526-09).

[0032] (1) Open field test Behavioral experiments were conducted in week 8 to investigate the effects of GMD intervention on cognitive and memory functions in mice. The Open Field Test (OFT) was used to assess the mice's spatial exploration abilities and anxiety-depression-like behaviors. Rodents' natural avoidance of open areas (approach-avoidance) and reduced exploration of central areas are typical manifestations of anxiety.

[0033] The experimental setup consisted of a square open-field box, with its bottom divided into a central area and a peripheral area. Before the formal test, mice were placed inside the box for 2 minutes to acclimatize to the environment, minimizing the interference of environmental variables (such as changes in odor and light) on their behavior. After the acclimatization period, the mice were placed in the central area for a 5-minute formal test. A behavior tracking system recorded the duration, distance, and trajectory of their activities in the central and peripheral areas in real time. After each test, the inside of the box was thoroughly wiped with 75% ethanol to eliminate any residual odor that could affect subsequent experimental results.

[0034] The effects of glyceryl decanoate (GMD) on the motor and spatial exploration abilities of mice fed a high-fat diet were measured in this experiment. Figure 1 As shown in the figure, a: representative movement trajectories of different groups in the open field; b: total activity distance in the open field; c: activity distance at the center of the open field; d: dwell time in the central area; e: percentage of time spent in the central area compared to the surrounding areas.

[0035] The results showed that the total open field activity distance of the HFD group ( Figure 1 (b) Distance between the center of the open field and the activity area ( Figure 1 c) Stay time in the central area ( Figure 1 (d) and the percentage of time spent in the central area compared to the surrounding areas ( Figure 1 Both (e) and (e) were significantly reduced, indicating that the high-fat diet significantly reduced the movement and spatial exploration abilities of mice. Meanwhile, the open field center movement distance (e) in the GMD group was significantly reduced. Figure 1 c) Stay time in the central area ( Figure 1 (d) and the percentage of time spent in the central area compared to the surrounding areas ( Figure 1 Both (e) increased significantly, indicating that GMD can improve its space exploration ability and anxiety-like behavior.

[0036] (2) New object recognition experiment In week 8, a Novel Object Recognition Test (NOR) was conducted to assess the spatial memory and cognitive function of mice. This test was performed sequentially over three consecutive days, divided into three phases: adaptation, training, and testing. Adaptation period (day 1): Mice freely explore a square open space box without objects to familiarize themselves with the experimental environment; Training period (day 2): Place two identical objects symmetrically inside the box (9 cm from the box wall) and allow the mice to explore freely for 5 minutes; Test period (day 3): Replace one of the old objects with a new object, place the mouse in the center of the box and let it explore freely for 5 minutes, and record the duration of its exploration of the new and old objects respectively.

[0037] After each test, the chamber and object surfaces were thoroughly cleaned with 75% ethanol to eliminate any residual odor that might interfere with subsequent experiments.

[0038] Discrimination Index = (New object exploration time – Old object exploration time) / (New object exploration time + Old object exploration time) Preference Index = New object exploration time / (New object exploration time + Old object exploration time).

[0039] The effects of glyceryl decanoate (GMD) on spatial cognition and memory in mice fed a high-fat diet were measured in this experiment. Figure 2 As shown in the figure, a: representative trajectories of new object recognition in different groups; b: new object recognition resolution index; c: new object recognition preference index.

[0040] The results showed that the discrimination index and preference index of the HFD group tended to decrease compared with the NCD group, indicating that a long-term high-fat diet weakened the mice's ability to consolidate memories of familiar objects, leading to failure in distinguishing between new and old objects. In contrast, the GMD group significantly improved the discrimination index and preference index, indicating that GMD can significantly improve cognitive dysfunction and memory decline in mice.

[0041] (3) Elevated cross maze test The elevated plus maze (EPM) test was conducted at week 8. The EPM test is used to assess anxiety-like behaviors in mice. In this experiment, the EPM test was used to evaluate the effect of GMD on HFD-induced anxiety-depression-like behaviors.

[0042] The experimental setup consisted of two open arms (without sidewalls) and two closed arms (with 15 cm high sidewalls) perpendicularly intersecting each other, with the entire setup 60 cm above the ground. During testing, mice were placed in the central area of ​​the maze, with their heads facing the open arms. The duration of time the mice spent on each arm and their movement trajectory were recorded continuously for 5 minutes. After each test, all arms and the central area of ​​the maze were thoroughly wiped with 75% ethanol to eliminate any odor residue that might interfere with subsequent experiments.

[0043] The effects of glyceryl decanoate (GMD) on anxiety-depressive-like behavior in mice fed a high-fat diet were measured in this experiment. Figure 3 As shown in the figure, a: representative motion trajectories of elevated cross mazes in different groups; b: percentage of open arm movement distance; c: ratio of open arm movement distance to closed arm movement distance; d: percentage of open arm dwell time; e: ratio of open arm movement time to closed arm movement time.

[0044] The results showed that the percentage of open arm movement distance in the HFD group ( Figure 3 (b) Open arm / closed arm travel distance ratio ( Figure 3 c) Percentage of open arm dwell time ( Figure 3 (d), open arm / closed arm activity time ratio ( Figure 3 The percentage of open arm movement distance (e) in the GMD group was significantly reduced, indicating that a high-fat diet can induce anxiety-depression-like behavior in mice, reflecting the enhanced avoidance of dangerous environments in HFD mice, consistent with the "approach-avoidance" instinct in rodents. Figure 3 (b) Open arm / closed arm travel distance ratio ( Figure 3 c) Percentage of open arm dwell time ( Figure 3 (d), open arm / closed arm activity time ratio ( Figure 3 The significant increases in both (e) and (e) indicate that GMD alleviates anxiety-like behaviors by reducing the generalization of fear memories and enhancing risk assessment capabilities.

[0045] (4) RNA extraction and real-time quantitative PCR At week 14, 30-50 mg of mouse brain tissue was added to Trizol, and RNA was extracted according to the reagent instructions. The RNA concentration and the absorbance ratio at 260 nm / 280 nm were measured using a Nanodrop spectrophotometer, and samples with an absorbance ratio between 1.8 and 2.0 were selected. The RNA was diluted to the same concentration according to experimental requirements. A reverse transcription system was prepared, and the RNA was reverse transcribed into cDNA using a PCR instrument.

[0046] The reverse transcription system includes: first step: Table 1

[0047] Step Two: Table 2

[0048] Reverse transcription PCR program settings: (1) Genomic DNA removal: After the reaction solution is prepared in the first step, gently mix it by pipetting and incubate at 42°C for 2 min.

[0049] (2) Preparation of reverse transcription reaction system: Add 5 × qRT SuperMix II directly to the reaction tube of step 1 and mix gently by pipetting. Place the reaction tube in a 50℃ water bath for 15 min, and then in an 85℃ water bath for 2 min. The product can be used immediately for qPCR reaction.

[0050] Prepare the experimental system according to the SYBR qPCR Master Mix instructions and set the expansion program accordingly. Record the Ct values, using GAPDH as an internal control, and follow the 2... - Ct Calculate relative gene expression. Primer sequences are shown in Table 3.

[0051] Table 3 RT-qPCR Primer Sequences

[0052] To determine whether GMD could alleviate neuroinflammation induced by HFD in obese mice, this experiment measured the mRNA expression levels of neuroinflammatory factors TNF-α, IL-1β, IL-6, COX-2, MCP-1, and iNOS in the brain. The effects of decanoic acid monoglyceride on neuroinflammation in mice fed a high-fat diet were also measured. Figure 4 As shown in the figure, af represents the mRNA expression levels of TNF-α, IL-1β, IL-6, COX-2, MCP-1, and iNOS, respectively. The results show that HFD can significantly upregulate TNF-α (… Figure 4 a) IL-1β Figure 4 (b) IL-6 Figure 4 c), COX-2 ( Figure 4 (d) MCP-1 ( Figure 4 (e), iNOS ( Figure 4 GMD significantly downregulates the mRNA expression level of TNF-α (f). Figure 4 a) IL-1β Figure 4 (b) IL-6 Figure 4 c), COX-2 ( Figure 4 (d) MCP-1 ( Figure 4 (e), iNOS ( Figure 4 The mRNA expression levels of (f) were observed. These results indicate that GMD can alleviate brain neuroinflammation by inhibiting the expression of neuroinflammatory factors (TNF-α, IL-1β, IL-6, MCP-1, iNOS).

[0053] Synaptic plasticity-related molecules play a crucial regulatory role in neurogenesis and brain function maintenance in mice. Among them, neurotrophic factors (such as BDNF, NT-3, and NT-4), postsynaptic density protein (PSD-95), and RNA-binding proteins (Fxr1 and Fxr2) are essential for maintaining the complex network of synaptic structure and function. Therefore, this experiment detected the mRNA expression of BDNF, NT-3, NT-4, and key synaptic structural proteins PSD-95, Fxr1, and Fxr2 in the brain. The effect of decanoic acid monoglyceride on synaptic plasticity in the brains of mice fed a high-fat diet is shown in Figure 5, where af represents the mRNA expression levels of BDNF, PSD-95, NT-3, NT-4, Fxr1, and Fxr2, respectively. The results showed that compared with the NCD group, the HFD group had significantly higher levels of BDNF (…). Figure 5 a) PSD-95 Figure 5 (b) NT-3 Figure 5 c), NT-4 ( Figure 5 d), Fxr1 ( Figure 5 (e), Fxr2 ( Figure 5 The mRNA expression of all six key genes (f) was significantly downregulated, while GMD significantly upregulated the expression levels of the above six key genes, indicating its potential to regulate synaptic plasticity in the mouse brain.

[0054] HFD activates the TLR4 / NF-κB pathway, and pro-inflammatory factors (TNF-α, IL-1β) inhibit hippocampal BDNF transcription. Impaired insulin signaling (PI3K / Akt) leads to decreased CREB phosphorylation, reduced PSD-95 synthesis, and damage to the structural stability of the postsynaptic compact area. Oxidative stress induced by HFD activates the JNK pathway and inhibits NT-3 / NT-4-dependent TrkB / TrkC receptor dimerization.

[0055] The above results indicate that GMD can alleviate neuronal damage to the brain caused by HFD. GMD can alleviate HFD diet-induced cognitive dysfunction by upregulating the expression of cognitive-related genes (PSD-95, Fxr1, Fxr2).

[0056] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. Use of capric acid monoglyceride in the preparation of a medicament for improving cognitive impairment.

2. Use according to claim 1, characterized in that, The capric acid monoglyceride is the only active ingredient in the medicament.

3. Use according to claim 1, characterized in that, The medicament comprises a therapeutically effective amount of capric acid monoglyceride or a pharmaceutically acceptable carrier or excipient.

4. Use according to claim 3, characterized in that, The dosage form of the medicament is a tablet, granules, oral liquid preparation, injection preparation or capsule preparation.

5. The use according to claim 1, characterized in that, The improvement of cognitive impairment comprises at least one of the following: (1) enhancing the memory ability of a cognitive impairment subject; (2) inhibiting the expression of brain neuroinflammatory factors in a cognitive impairment subject to alleviate brain neuroinflammation; (3) increasing the expression level of neurotrophic factors and cognitive-related genes in the brain tissue of a cognitive impairment subject.

6. Use according to claim 5, characterized in that, The neuroinflammatory factors include at least one of TNF-α, IL-1β, IL-6, MCP-1 and iNOS.

7. Use according to claim 5, characterized in that, The neurotrophic factors include at least one of BDNF, NT-3 and NT-4; the cognitive-related genes include at least one of PSD-95 gene, Fxr1 gene and Fxr2 gene.

8. The use according to claim 1, characterized in that, The cognitive impairment includes at least one of the following behavioral disorders: memory disorder, visual space disorder, computational ability disorder.

9. Use of capric acid monoglyceride in the preparation of a health food for enhancing memory.

10. Use according to claim 9, characterized in that, The health food comprises capric acid monoglyceride or a food-acceptable carrier or excipient.

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