Use of daucosterol in the preparation of a medicament for treating delirium-related cognitive impairment

Abstract

The application discloses application of daucosterol in preparation of a medicine for treating delirium-related cognitive impairment. The application first discovers that daucosterol intragastric administration shows a significant improvement in cognitive function on a drug-induced delirium mouse model, and compared with a solvent group, daucosterol can significantly reduce cognitive impairment. In vitro experiments show that daucosterol can activate nuclear receptors ROR alpha and ROR gamma, up-regulate the expression of a target gene E4BP4, and then inhibit Mapk1 and Mapk3, which are genes related to cognitive impairment, to regulate cognitive impairment. In conclusion, daucosterol plays a role in improving delirium through a ROR alpha / gamma-E4BP4-Mapk1 / 3 pathway. In addition, daucosterol shows good compatibility when combined with other delirium medicines. Daucosterol is a natural ingredient, has obvious cost advantages in production, and has excellent safety.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of lentinan in drugs for the prevention and / or treatment of delirium-related cognitive impairment. Background Technology

[0002] Cognitive disorder refers to a decline or impairment in an individual's cognitive abilities, including thinking, memory, learning, comprehension, concentration, and decision-making. It affects an individual's processing and acquisition of information and can negatively impact daily functioning and life. Cognitive disorder has a significant impact on an individual's daily living functions, independence, and social interaction abilities. It can be caused by a variety of factors, including delirium, Alzheimer's disease, Parkinson's disease, and cerebrovascular diseases. Cognitive disorder is associated with several risk factors, such as hypertension, cardiovascular disease, diabetes, obesity, smoking, lack of exercise, low education level, and genetic factors. Given the diverse and multifactorial etiologies of cognitive disorder, its pathogenesis may involve multiple neurobiological mechanisms, including neuroinflammation, altered brain energy metabolism, cerebrovascular dysfunction, neurotransmitter disorders, and weakened neural network connectivity.

[0003] Delirium, also known as acute brain syndrome, is a severe neuropsychiatric syndrome characterized primarily by acute cognitive and attentional deficits. The prevalence of severe cases can be as high as 60–80%. Delirium is associated with prolonged hospital stays, cognitive and motor impairments, and increased mortality. Risk factors for delirium include acute medical conditions (such as sepsis and liver failure), trauma (such as fractures and severe burns), surgery, and medications (such as benzodiazepines, antihistamines, and opioids). Developing drugs to treat cognitive impairment associated with diseases such as delirium has significant clinical value.

[0004] Lysionotin (Formula 1) is a natural flavonoid compound primarily derived from the herbaceous plant *Primulina* of the Gesneriaceae family. This compound is a trimethoxylated member of the flavonoid family. Studies have revealed a variety of pharmacological effects of lysionotin, including antibacterial, anti-inflammatory, antihypertensive, and free radical scavenging effects. In the intervention of lung diseases, lysionotin has been widely used in combination therapy with other drugs or herbs. In terms of antibacterial activity, lysionotin has been shown to exert its effects by inhibiting the α-toxin produced by *Staphylococcus aureus*. Furthermore, this compound has also shown anti-inflammatory efficacy in rat models, particularly in a carrageenan-induced paw edema model. Notably, although some disease intervention effects of lysionotin have been discovered, its potential in the treatment of cognitive impairment has not been fully investigated.

[0005]

[0006] The nuclear receptor superfamily is a group of ligand-dependent transcription factors that play important regulatory roles in a variety of physiological processes. Retinol-associated orphan receptors (RORs) are an important subfamily of this superfamily, consisting of three members: RORα, RORβ, and RORγ. These receptors regulate the promoter activity of target genes by binding to specific ROR response elements (ROREs), where each RORE consists of a core motif AGGTCA and a preceding 5-bp A / T-rich sequence. RORs can interact with specific co-repressors and co-activators to achieve cell- and gene-specific regulation, thereby modulating the expression and function of downstream genes. Within the ROR subfamily, RORα and RORγ exhibit widespread expression patterns in various tissues, including the cerebellum, liver, thymus, skeletal muscle, skin, lung, adipose tissue, and kidney. RORγ is highly expressed in the thymus and plays an important role in immune regulation. In contrast, RORβ is mainly expressed in specific regions of the central nervous system, such as the retina and pineal gland.

[0007] E4BP4 (NFIL3) is a direct target of RORs (Reactive Oocytes) and participates in various physiological processes, including transcriptional control of the circadian rhythm, neuronal growth and survival, osteoblast function, and regulation of ovulation. In the pathophysiology of delirium, E4BP4 exhibits a crucial regulatory role. Deletion of E4BP4 in the mouse hippocampus exacerbates cognitive impairment and increases the expression of key cognitive impairment-related enzymes Mapk1 and Mapk3. E4BP4 regulates post-delirium cognitive impairment at the transcriptional level by directly binding to D-box response elements on the promoters of cognitive impairment-related genes (Mapk1 and Mapk3). In conclusion, E4BP4 not only plays a crucial role in immune regulation and various physiological processes but is also an important regulator of cognitive impairment. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings and deficiencies of the prior art and to provide the application of lentinan in the preparation of drugs for the treatment and / or prevention of cognitive impairment.

[0009] The objective of this invention is achieved through the following technical solution:

[0010] The use of lentinan in the preparation of drugs for the treatment and / or prevention of cognitive impairment.

[0011] Furthermore, the cognitive impairments mentioned include, but are not limited to, cognitive impairments caused by delirium, neurodegenerative diseases, and cerebrovascular diseases.

[0012] Furthermore, the application is the use of lentinan in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment.

[0013] Furthermore, the lentinan has the effect of regulating the expression of cognitive impairment regulatory factors.

[0014] Furthermore, the regulatory factors include the negative regulatory factor E4BP4 and the positive regulatory factors Mapk1 and Mapk3.

[0015] Furthermore, the romolysin upregulates the expression of the negative regulator E4BP4 and downregulates the expression of the positive regulators Mapk1 and Mapk3 by activating nuclear receptors RORα and RORγ.

[0016] Furthermore, the mass percentage of the genistein in the drug is 0.01–99.5%.

[0017] In the above applications, the physalisin also includes pharmaceutically acceptable inorganic or organic acid salts of physalisin, including but not limited to hydrochloride, phosphate, sulfate, acetate, maleate, methanesulfonate, fumarate, citrate, benzenesulfonate, methylbenzenesulfonate, or tartrate.

[0018] In the above applications, the available dosage forms of the drug include various oral and injectable dosage forms, including but not limited to tablets, capsules, granules, powders, powder injections, pills, emulsions, suspensions, solutions, sprays, patches, or sublingual tablets.

[0019] In the above applications, the drug may also include various pharmaceutically acceptable excipients / components, including but not limited to coating materials, solvents, solubilizers, binders, stabilizers, antioxidants, pH adjusters, flavoring agents, etc., with particular preference given to various excipients / components for oral dosage forms.

[0020] In the above applications, the medication may include other Chinese and Western medicines that prevent and / or treat cognitive impairment, or may be used in combination with treatments such as surgery.

[0021] In the above applications, the structural formula of the physalisin is shown in Formula 1:

[0022]

[0023] This invention utilizes an animal model of delirium induced by LPS combined with midazolam. Benzodiazepines (midazolam) alter human sleep patterns and increase the risk of delirium, while inflammation can also induce delirium. Use of midazolam or LPS in mice resulted in significant reductions in cognitive function, motor activity, and memory function, exhibiting a delirium-like phenotype similar to that in humans. Therefore, this model aligns with current phenomenological models of human delirium.

[0024] The present invention has the following advantages and effects compared with the prior art:

[0025] In this invention, gavage administration of lentinan showed a significant improvement in cognition in a mouse model of drug-induced delirium. Compared with the solvent group, lentinan significantly reduced cognitive impairment. In vitro experiments showed that lentinan can activate nuclear receptors RORα and RORγ, upregulate the expression of its target gene E4BP4, and thereby inhibit cognitive impairment-related genes Mapk1 and Mapk3, thus regulating cognitive dysfunction. In summary, lentinan exerts its effect in improving delirium through the RORα / γ-E4BP4-Mapk1 / 3 pathway.

[0026] Furthermore, lentinan exhibits good compatibility when combined with other delirium medications.

[0027] Rock bean extract is a natural ingredient derived from plants such as rockwort. It has significant cost advantages in production and exhibits excellent safety profiles. Attached Figure Description

[0028] Figure 1 This is the chemical structure diagram of lentinan.

[0029] Figure 2 The figure shows the results of a study on the effect of lentinan on object preference in a novel object recognition experiment in delirious mice.

[0030] Figure 3 The figure shows the results of a study on the effect of follicularin on the exploration time in a novel object recognition experiment in delirious mice.

[0031] Figure 4 Figure 1 shows the results of a study on the effects of lentinan on alternating Y-maze behavior in delirious mice.

[0032] Figure 5 The figure shows the results of the study on the activation effect of lentinan on RORα / γ / β.

[0033] Figure 6 Figure showing the results of a study on the effect of lentinan on the ROR target gene E4bp4.

[0034] Figure 7 This is a figure showing the results of a study on the effects of lentinan on the cognitive impairment-related genes Mapk1 and Mapk3. Detailed Implementation

[0035] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0036] Example 1: Establishment of a mouse model of delirium

[0037] Healthy male mice of the C57BL / 6 strain, aged 8–12 weeks (purchased from Guangzhou Zitangjin Biotechnology Co., Ltd.), were used. All mice were housed in an SPF-grade environment with 12 hours of light / 12 hours of darkness (light period from 7:00 AM to 7:00 PM) at a temperature of 22–25°C. We first acclimatized all mice to the environment for 7 days, providing standard feed and purified water. After the acclimatization period, the mice were randomly divided into two groups: a normal control group and a delirium model group (n=10 in the normal control group and n=40 in the delirium model group).

[0038] Normal control group: fed with standard feed, and simultaneously injected intraperitoneally with blank solvent (distilled water).

[0039] Delirium model group: LPS (200 μg / kg) plus midazolam (10 mg / kg) was administered intraperitoneally once daily for 3 consecutive days to induce delirium-like syndrome. Mice were divided into a solvent control group, a prophylactic administration group, and a therapeutic administration group, with 10 mice in each group.

[0040] (1) Solvent control group: Blank reagent (distilled water) was administered by gavage during modeling;

[0041] (2) Prophylactic administration group: 3 days before the start of modeling, lentinan was administered by gavage. The administration of lentinan was divided into a low concentration group (10 mg / kg) and a high concentration group (50 mg / kg).

[0042] (3) Therapeutic administration group: One day after modeling, lentinan was administered by gavage for 3 days. The administration of lentinan was divided into a low concentration group (10 mg / kg) and a high concentration group (50 mg / kg).

[0043] 72 hours later, the mice underwent behavioral tests. The mice were then euthanized to collect plasma and hippocampal samples for corticosterone measurement (ELISA), qPCR, and / or Western blot analysis.

[0044] Example 2: Study on the therapeutic effect of lentinan on cognitive impairment

[0045] Both prophylactic and therapeutic administration of lentinan significantly improved the cognitive state of delirium-inducing mice, showing a significant difference compared to the solvent group. After administration of lentinan, delirium-inducing mice exhibited a significantly higher preference for new objects than for old objects in the new object recognition test, with high concentrations of lentinan showing a more pronounced therapeutic effect, comparable to normal control mice. Figure 2 Regarding exploration time, the delirium model mice treated with lentinan also showed similar results to the normal control mice, both spending more time exploring new objects than the solvent-treated delirium model mice. Figure 3This indicates that fosetyl-methyl can restore memory and recognition abilities in delirium-inducing mice. In the Y-maze experiment, the fosetyl-methyl group significantly increased the number of alternations in delirium-inducing mice. Figure 4 ).

[0046] The above experiments show that lentinan can improve memory and cognition in delirious mice.

[0047] Example 3: Study on the activating effect of lentinan on nuclear receptors RORα / β / γ

[0048] The agonistic effect of lentinan on the nuclear receptor RORα was investigated using a dual-luciferase reporter gene assay. In the Gal4 co-transfection assay, only the specific ligands of RORα / γ / β could bind to the ligand-binding domain (LBD) to activate gene transcription. Microglia (BV2 cells, purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences) were co-transfected with the GAL4-NR LBD structure (purchased from BioNTech), pGL4.35 [luc2P / 9XGAL4UAS / Hygro] vector, and pRL-TK vector. On the second day, the cells were incubated with lentinan (10 μM) or dimethyl sulfoxide (solvent control) for 24 hours. The activity of luciferase was then measured using a dual-luciferase reporter gene assay kit (Promega, Madison, WI) and a Glomax 20 / 20 luminometer. In addition, microglia were incubated with lentinan (10 μM) or dimethyl sulfoxide (solvent control) for 24 hours, and the expression of target genes E4bp4, Mapk1 and Mapk3 was measured by qPCR.

[0049] The results showed that dual-luciferase reporter gene assays demonstrated that lentin significantly activated RORα and RORγ activities, but had no activating effect on RORβ. Figure 5 Furthermore, lentinan can increase the expression of the RORα / γ target gene E4bp4 (a negative regulator of delirium and cognitive impairment). Figure 6 ), reducing the expression of delirium-related genes Mapk1 and Mapk3 ( Figure 7 In summary, lentinan exerts its effect on improving post-delirium cognitive impairment through the RORα / γ-E4BP4-Mapk1 / 3 pathway.

[0050] 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 lentinan in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment.

2. The use of lentinan according to claim 1 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: The aforementioned lentinan has the effect of regulating the expression of negative regulatory factors of cognitive impairment.

3. The use of lentinan according to claim 2 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: The negative regulatory factors mentioned include E4BP4; The lentinan upregulates the expression of the negative regulator E4BP4 by activating nuclear receptors RORα and RORγ.

4. The use of lentinan according to claim 3 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: The mass percentage of the lentinan in the drug is 0.01 to 99.5%.

5. The use of lentinan according to claim 1 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: In applications, the genistein also includes pharmaceutically acceptable inorganic or organic acid salts of genistein.

6. The use of lentinan according to claim 5 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: In applications, the genistein also includes pharmaceutically acceptable hydrochloride, phosphate, sulfate, acetate, maleate, methanesulfonate, fumarate, citrate, benzenesulfonate, methylbenzenesulfonate, or tartrate of genistein.

7. The use of lentinan according to claim 1 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: In application, the dosage form of the drug includes tablets, capsules, granules, powders, powder injections, pills, emulsions, suspensions, solutions, sprays, or patches.

8. The use of lentinan according to claim 1 in the preparation of drugs for the treatment and / or prevention of delirium-related cognitive impairment, characterized in that: In application, the drug also includes pharmaceutically acceptable coating materials, solvents, solubilizers, binders, stabilizers, antioxidants, pH adjusters, or flavoring agents.

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