Use of lysophosphatidylcholine (16:0) in treatment and prevention of alzheimer's disease and postoperative cognitive dysfunction

Abstract

This invention relates to the field of biomedicine, providing the application of lysophosphatidylcholine (16:0) in the treatment and prevention of Alzheimer's disease and postoperative cognitive impairment. Experimental results suggest that LyPC (16:0) is an important protective metabolite regulating cognitive function, and its deficiency is a significant factor in the occurrence and development of AD and SICI, providing a novel endogenous target for the intervention of cognitive impairment diseases. Exogenous administration of LyPC (16:0) can effectively improve cognitive function in AD and SICI models, reverse pathological changes such as decreased hippocampal neuronal excitability and synaptic dysfunction, inhibit glial cell activation, reduce neuroinflammation levels, and maintain normal serum LyPC (16:0) levels, reducing the risk of AD and the risk of cognitive decline caused by surgical trauma. This invention clarifies the dual therapeutic and preventive effects of LyPC (16:0) on AD and SICI, provides multiple safe and effective supplementation pathways, has a clear mechanism of action and high safety, and provides a novel strategy for the clinical intervention of cognitive impairment-related diseases.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine, functional food and pharmaceutical technology, and to a new use of lysophosphatidylcholine (16:0) (LyPC (16:0)), specifically to its use in the preparation of drugs, foods and health products for the treatment and prevention of Alzheimer's disease (AD) and postoperative cognitive impairment (SICI, including postoperative delirium POD). Background Technology

[0002] Cognitive impairment is a major public health problem facing aging societies. Among them, Alzheimer's disease (AD) and postoperative cognitive impairment (SICI) are two types of cognitive disorders that are highly prevalent and have serious consequences, placing a heavy burden on patients, families and society.

[0003] Alzheimer's disease (AD) is a progressive neurodegenerative disease of the central nervous system. Its core pathological features include β-amyloid (Aβ) plaque deposition in the brain, hyperphosphorylation of tau protein in neurons, accompanied by hyperactivation of glial cells in the hippocampus, severe neuroinflammation, decreased neuronal excitability, damage to synaptic structure and function, and a significant decrease in dendritic spine density. Clinically, it manifests as progressive decline in spatial learning and memory abilities, ultimately leading to loss of self-care ability. Currently, clinical treatments for AD can only temporarily relieve symptoms and cannot reverse the disease progression. Furthermore, they have single targets and limited efficacy, lacking effective means to fundamentally intervene in the occurrence and development of the disease.

[0004] Postoperative cognitive impairment (SICI) is a common complication after surgery, especially in elderly patients. Postoperative delirium (POD) is the most typical manifestation, referring to attention deficit, altered consciousness, and cognitive decline that occur within one week after surgery (usually 24-72 hours). Its pathogenesis is closely related to surgical trauma, anesthesia-induced hippocampal neuroinflammation, glial cell activation, blood-brain barrier disruption, and neuronal dysfunction. Currently, there are no specific drugs for the prevention and treatment of SICI in clinical practice. Interventions can only be provided through perioperative care and management of underlying diseases, and the high incidence of postoperative cognitive impairment has not been effectively reduced.

[0005] Recent studies have shown a close bidirectional relationship between peripheral blood metabolites and brain function regulation, and the disorder of circulating metabolites is one of the important causes of cognitive impairment. Lysophosphatidylcholine, as a class of endogenous lipid biomolecules, has traditionally been considered to have elevated levels and pathogenicity in diseases such as cardiovascular disease, diabetes, and stroke, but whether it has a link with neurodegenerative diseases has not yet been reported. Summary of the Invention

[0006] This invention, based on the aforementioned research, explores the application of lysophosphatidylcholine (16:0) in the treatment and prevention of Alzheimer's disease and postoperative cognitive impairment. Extensive clinical sample testing and animal experiments revealed for the first time that lysophosphatidylcholine (16:0) (LyPC (16:0)) showed a significant downregulation trend in the serum of AD, SICI patients, and corresponding animal models. Furthermore, the level of this metabolite was significantly positively correlated with cognitive function scores such as the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA), and negatively correlated with age. This suggests that LyPC (16:0) is an important protective metabolite regulating cognitive function, and its deficiency is a crucial factor in the occurrence and development of AD and SIC, providing a novel endogenous target for the intervention of cognitive impairment diseases.

[0007] Exogenous administration of LyPC(16:0) via intravenous or intraperitoneal injection, or supplementation via diet or oral administration, can effectively improve cognitive function in AD and SICI models, reverse pathological changes such as decreased hippocampal neuronal excitability and synaptic dysfunction, inhibit glial cell activation, reduce neuroinflammation levels, and maintain normal serum LyPC(16:0) levels, thus reducing the risk of AD and cognitive decline caused by surgical trauma. This invention clarifies the dual therapeutic and preventative effects of LyPC(16:0) on AD and SICI, provides multiple safe and effective supplementation routes, has a clear mechanism of action and high safety, and offers a novel strategy for clinical intervention in cognitive impairment-related diseases, with promising industrialization prospects and clinical application value.

[0008] Based on this, this invention develops the application of LyPC (16:0) in the treatment and prevention of AD and SICI, providing a safe and effective supplementary approach. The technical solution to be protected by this invention is as follows:

[0009] In a first aspect, the present invention provides the use of lysophosphatidylcholine (16:0) (LyPC (16:0), CAS No.: 17364-16-8) in the preparation of a medicament for treating cognitive impairment, wherein the cognitive impairment includes Alzheimer's disease and postoperative cognitive impairment.

[0010] The drug, by supplementing LyPC (16:0), improves cognitive functions such as spatial learning and memory in AD patients, and alleviates AD-related pathological damage, including decreased hippocampal neuronal excitability, synaptic transmission dysfunction, excessive glial cell activation, neuroinflammation, and decreased dendritic spine density. Supplementing LyPC (16:0) also improves postoperative learning and memory abilities and alleviates SICI-related pathological damage, including hippocampal neuronal dysfunction, glial cell activation, neuroinflammation, and decreased dendritic spine density.

[0011] In a second aspect, the present invention provides the use of lysophosphatidylcholine (16:0) in the preparation of food or health products for the prevention of cognitive impairment, wherein the cognitive impairment includes Alzheimer's disease and postoperative cognitive impairment.

[0012] When used to prevent Alzheimer's disease (AD), LyPC (16:0) can be supplemented through diet and oral administration to maintain normal levels of this metabolite in the body's serum, inhibit age-related neuroinflammation, maintain the integrity of hippocampal synaptic structure, and reduce the risk of AD in the elderly.

[0013] When used to prevent postoperative cognitive impairment, LyPC(16:0) can be supplemented preoperatively and continuously postoperatively to maintain stable serum levels of this metabolite, inhibit neuroinflammation caused by surgical trauma, and reduce the risk of postoperative cognitive decline in surgical patients, especially elderly patients. Supplementation methods include injection, diet, and oral administration.

[0014] Preferably, the drug, food, or health food uses lysophosphatidylcholine (16:0) as the sole active ingredient or in combination with other active ingredients.

[0015] Regarding the formulation, the drug is selected from injectable or oral formulations, such as tablets, capsules, and traditional Chinese medicine pills. The injectable formulation uses physiological saline or physiological buffer solution as a solvent and is prepared with a pharmaceutically acceptable carrier. The route of administration includes one or more of tail vein injection, intraperitoneal injection, and subcutaneous injection; the concentration of LyPC (16:0) in the drug is 1 μg / ml to 60 μg / ml, with a preferred dosage of 20 mM / time or 200 μL / time, administered twice weekly for a continuous period of not less than 6 weeks.

[0016] The daily supplemental dose of LyPC (16:0) in the food or health product is 0.1mg~10mg / kg body weight. It can be prepared by adding natural raw materials rich in LyPC (16:0) or by directly adding purified LyPC (16:0). It can be formulated into oral preparations, capsules, tablets, functional foods, etc. with the help of food science-acceptable excipients.

[0017] Preferably, the lysophosphatidylcholine (16:0) is selected from natural raw materials rich in LyPC (16:0) or isolated and purified LyPC (16:0).

[0018] In a third aspect, the present invention provides a composition for treating or preventing Alzheimer's disease or postoperative cognitive impairment, wherein LyPC (16:0) is the sole or main active ingredient, supplemented with a pharmaceutically acceptable carrier or a food-acceptable excipient.

[0019] According to the application scenario, it is made into injections, oral preparations, capsules, tablets, and functional foods. The injections are suitable for clinical treatment, while the oral preparations, capsules, tablets, and functional foods are suitable for daily prevention.

[0020] Furthermore, depending on the actual situation, the above composition may be combined with other drugs for treating AD or SICI, administered simultaneously, sequentially, or alternately.

[0021] In a fourth aspect, the present invention provides the use of a reagent for detecting lysophosphatidylcholine (16:0) levels in the preparation of a diagnostic or prognostic kit for Alzheimer's disease or postoperative cognitive impairment.

[0022] This invention, through clinical sample testing and animal model experiments, confirms that the serum levels of LyPC (16:0) in AD and SICI patients and corresponding animal models are significantly downregulated, and that the level of this metabolite is positively correlated with cognitive function scores and negatively correlated with age. Based on this, it can be inferred that LyPC (16:0) can serve as a diagnostic biomarker or drug screening biomarker for AD and SICI.

[0023] By regularly detecting the expression level of LyPC (16:0) in peripheral blood, the risk of AD in the elderly can be predicted or the prognosis of AD patients after treatment can be assessed; by detecting the expression level of LyPC (16:0) in peripheral blood after surgery, the risk of SICI in postoperative patients can be assessed or the treatment effect can be predicted.

[0024] The method for drug screening using LyPC (16:0) as a biomarker is as follows:

[0025] (A) Treating AD or SICI animal models with candidate substances;

[0026] (B) Detection of LyPC (16:0) levels in peripheral blood of animal models;

[0027] (C) If the LyPC (16:0) level is higher than the expression level in peripheral blood before drug administration after treatment with the candidate substance, it indicates that the candidate substance has the effect of alleviating AD or SICI.

[0028] In a sixth aspect, the present invention provides a diagnostic or prognostic kit for Alzheimer's disease or postoperative cognitive impairment, containing a reagent for detecting the level of lysophosphatidylcholine (16:0).

[0029] The beneficial protections and effects of this invention are as follows:

[0030] 1. A new application of LyPC (16:0) is clearly defined for the first time, filling a gap in the field: The endogenous metabolite LyPC (16:0) has been confirmed for the first time to have both therapeutic and preventive effects on AD and SICI, providing a new endogenous target for the intervention of cognitive impairment diseases and breaking the dilemma of existing AD treatment targets being single and SICI lacking specific intervention methods.

[0031] 2. Clear mechanism of action and significant intervention effect: LyPC (16:0) works synergistically through multiple targets, which can simultaneously improve the excitability and synaptic function of hippocampal neurons, inhibit glial cell activation, reduce neuroinflammation, and protect the integrity of synaptic structure. It has a significant effect on improving cognitive function and alleviating pathological damage in AD and SICI, and this has been fully validated in animal models.

[0032] 3. Offers multiple supplementary routes to suit different clinical scenarios: In response to different treatment and prevention needs, it offers multiple supplementary routes such as injection, diet, and oral administration. Injectable preparations are suitable for the precise treatment of AD and SICI patients in clinical practice, while oral preparations and functional foods are suitable for daily prevention in the elderly and high-risk surgical populations, meeting the intervention needs of different populations and scenarios.

[0033] 4. High safety and no obvious toxic side effects: LyPC (16:0) is a naturally occurring endogenous lipid metabolite in the body. Exogenous supplementation is only to make up for the body's deficiency and will not cause metabolic disorders. Animal experiments have shown that it has no significant effect on the movement ability of mice, and its safety is far higher than that of traditional chemical drugs.

[0034] 5. Wide range of raw material sources, easy to industrialize and promote: LyPC (16:0) can be prepared on a large scale through chemical synthesis, extraction of natural raw materials, etc. Its preparation process for injections, oral preparations and functional foods are all existing mature processes. It can be industrialized through conventional pharmaceutical and food processing equipment, with controllable production costs and strong industrial applicability.

[0035] 6. Broad market prospects: The LyPC (16:0) and related compositions of this invention can be widely used in hospital clinical practice (treatment of AD and SICI patients), health care products industry (AD prevention in the elderly population and SICI prevention in high-risk surgical populations), pharmaceutical industry (research and development and production of AD and SICI treatment drugs), and many other fields. The market demand is large and the application prospects are broad.

[0036] In summary, this invention has significant industrial applicability and market value, and can be industrialized and promoted to generate good economic and social benefits. Attached Figure Description

[0037] The present disclosure will be further described below with reference to the accompanying drawings, which are shown only for illustrating the embodiments of the present disclosure and are not intended to limit the scope of the present disclosure.

[0038] Figure 1 The following graphs show the correlation between serum metabolic profiles and cognitive function: A is the experimental flowchart of serum non-targeted mass spectrometry detection in healthy young adults and the elderly; B is a comparison of baseline characteristics between the two groups; C and D are comparisons of Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) scores between the two groups; E is a Spearman correlation analysis of MMSE / MoCA scores with age; F is a principal component analysis (PCA) graph of serum metabolites in the two groups, showing a significant separation between the two metabolic profiles; G is the screening results of differentially expressed metabolites between the two groups (|logFC|>0.25, adjusted-P<0.05), with a total of 39 differentially expressed molecules screened; H and I are heatmaps of 20 downregulated and 19 upregulated differentially expressed molecules; J is a comparison of LyPC (16:0) expression differences between the two groups, representing the most significant difference; K is a Spearman correlation analysis of the 39 differentially expressed molecules with MMSE, MoCA, and age; LN Spearman correlation analysis plot of LyPC (16:0) with age, MoCA, and MMSE shows that LyPC (16:0) is negatively correlated with age and positively correlated with cognitive score.

[0039] Figure 2The validation results of LyPC (16:0) downregulation in patients with cognitive impairment and corresponding animal models are shown in the following figures: A is a comparison of baseline characteristics among young adults, the elderly, and AD patients (n=6 / group); B is a comparison of serum LyPC (16:0) levels among the three groups, showing AD patients < healthy elderly < healthy young adults; C is a comparison of serum LyPC (16:0) levels between wild-type (WT) and AD mice; DF is a Spearman correlation analysis of LyPC (16:0) with age, MoCA, and MMSE; G is a comparison of baseline characteristics between non-postoperative delirium (Non-POD) and postoperative delirium (POD) patients (n=6 / group); H is a comparison of preoperative and postoperative serum LyPC (16:0) levels between Non-POD and POD patients; I is a comparison of LyPC (16:0) levels before and after surgery in POD and Non-POD patients. Comparison of level change values ​​(delta); J is a comparison of serum LyPC (16:0) levels between the control group and SICI mice; K is a comparison of plasma LyPC (16:0) levels between the control group and SICI mice; J and K both confirm that LyPC (16:0) is significantly downregulated in the SICI model.

[0040] Figure 3 The following diagrams show the experimental results of peripherally supplemented LyPC (16:0) improving cognitive and synaptic function in AD mice: A is a schematic diagram of the experimental process for AD mice treated with WT, AD, and LyPC (16:0); B and C are the results of the open field and rotarod tests for the three groups of mice (n=10 / group), showing no difference in motor ability among the three groups; D is the result of the five-day Morris water maze (MWM) training experiment for the three groups of mice, showing that the learning ability of the treatment group is significantly better than that of the AD model group; E is the representative movement trajectory diagram of the three groups of mice on the MWM test day; F and G are the statistics of the total swimming distance and platform crossing number of the three groups of mice on the MWM test day; H is the result of the new object recognition experiment for the three groups of mice; I is the result of the conditioned fear memory experiment for the three groups of mice; J is the representative trajectory diagram of the hippocampal pyramidal cell discharge characteristics of the three groups of mice under 50pA current injection (WT n=10, AD n=10, treatment group n=11); K The graph shows a comparison of the firing frequency of action potentials (AP) in the three groups of mice; L is a typical trajectory of small excitatory postsynaptic currents (mEPSC) in hippocampal neurons of the three groups of mice (WT n=12, AD n=12, treatment group n=13); M and N are comparison graphs of the amplitude and frequency of mEPSC in the three groups of mice; FN both confirmed that LyPC (16:0) can improve cognitive function and synaptic function in AD mice.

[0041] Figure 4 The following figures show the experimental results of peripherally supplemented LyPC (16:0) reducing glial cell activation, neuroinflammation, and dendritic spine density in AD mice: AD is a comparison of the average fluorescence intensity of MX04 in the CA1 and DG regions of the hippocampus of the three groups of mice (n=13 / group), showing no significant difference in Aβ plaques; EJ is an immunofluorescence staining image of IBA1 & 6E10 in the CA1 and DG regions of the hippocampus of the three groups of mice and a statistical count of microglia related to Aβ plaques (n=6 / group); KP is an immunofluorescence staining image of GFAP & 6E10 in the CA1 and DG regions of the hippocampus of the three groups of mice and a statistical count of astrocytes related to Aβ plaques (n=6 / group); QS is a comparison of the levels of inflammatory factors TNF, IL-1β, and IL-6 in the hippocampus of the three groups of mice (n=6 / group); T is a comparison of the dendritic spine density in the hippocampus of the three groups of mice (n=6 / group), scale bar = Both 100 μm and ET studies confirmed that LyPC (16:0) could reduce glial cell activation and neuroinflammation in AD mice and increase dendritic spine density.

[0042] Figure 5 The following figures illustrate the experimental results of improving cognitive function, synaptic function, and reducing neuroinflammation in SICI mice after intravenous injection of LyPC (16:0): A is a schematic diagram of the experimental process for the control group, SICI group, and LyPC (16:0)-treated SICI mice; BD shows the average movement speed, central region dwell time, and representative movement trajectory diagrams of the three groups of mice in the open field experiment (n=6 / group), showing no difference in movement ability; E shows the results of the conditioned fear memory experiment of the three groups of mice; F shows the results of the four-day training experiment in the Barnes maze of the three groups of mice; G shows the percentage of time spent in the target quadrant of the three groups of mice on the Barnes maze test day (n=8); H shows the representative movement trajectory diagrams of the three groups of mice on the Barnes maze test day; I shows the representative trajectory diagrams of hippocampal pyramidal cell discharge characteristics of the three groups of mice under 50pA current injection (control group n=12, SICI group n=10, treatment group n=12); J shows the comparison of action potential (AP) firing frequency of the three groups of mice; K Typical mEPSC trajectories of hippocampal neurons in three groups of mice (control group n=13, SICI group n=13, treatment group n=12); L and M are comparison diagrams of mEPSC amplitude and frequency in the three groups of mice; EM both confirmed that LyPC (16:0) can improve cognitive function and synaptic function in SICI mice.

[0043] Figure 6The experimental results of intravenous injection of LyPC (16:0) in reducing glial cell activation and neuroinflammation in SICI mice are shown in the following figures: AF shows the immunofluorescence staining of IBA1 in the CA1 and DG regions of the hippocampus of the three groups of mice and the statistical count of microglia (n=6 / group); GK shows the immunofluorescence staining of GFAP in the CA1 and DG regions of the hippocampus of the three groups of mice and the statistical count of astrocytes (n=6 / group); LN shows the comparison of the levels of inflammatory factors TNF, IL-1β, and IL-6 in the hippocampus of the three groups of mice (n=6 / group); O shows the comparison of the dendritic spine density in the hippocampus of the three groups of mice (n=6 / group), scale bar = 100μm; AO shows that LyPC (16:0) can reduce glial cell activation and neuroinflammation in SICI mice and increase dendritic spine density. Detailed Implementation

[0044] The following examples and experimental cases further illustrate the present invention and should not be construed as limiting the invention. The examples do not include detailed descriptions of conventional methods, such as methods for constructing vectors and plasmids, methods for inserting genes encoding proteins into vectors and plasmids, or methods for introducing plasmids into host cells. Such methods are well known to those skilled in the art and have been described in numerous publications.

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention, and the preferred embodiments and materials described herein are for illustrative purposes only.

[0046] This invention provides a novel application of lysophosphatidylcholine (16:0) (LyPC (16:0)), clarifying its significant effects in the treatment and prevention of Alzheimer's disease (AD) and postoperative cognitive impairment (SICI), elucidating its mechanism of action, and providing multiple safe and effective supplementation routes such as injection, diet, and oral administration. This invention offers new strategies and targets for the clinical prevention and treatment of AD and SICI, filling the gap in the field of endogenous lipid metabolite intervention for cognitive impairment.

[0047] The mechanisms by which LyPC (16:0) exerts its therapeutic and preventive effects are as follows: (1) it enhances the excitability of hippocampal neurons, increases the frequency of action potential firing, improves synaptic transmission function, and increases the frequency and amplitude of microexcitatory postsynaptic currents; (2) it inhibits the overactivation of microglia and astrocytes and reduces the number of plaque-associated glial cells; (3) it reduces the protein expression levels of inflammatory factors TNF, IL-1β, and IL-6 in hippocampal tissue and alleviates neuroinflammation; (4) it increases the density of dendritic spines in the hippocampus and improves and maintains the integrity of synaptic structure.

[0048] In a preferred embodiment of the present invention, through clinical sample testing, animal model construction, and a series of in vivo experiments, the therapeutic and preventive efficacy of LyPC (16:0) against AD and SICI was systematically demonstrated, and its mechanism of action was clarified. The specific technical solution is as follows:

[0049] 1. LyPC (16:0) was significantly downregulated in individuals and models of cognitive impairment.

[0050] Serum samples from healthy young adults, healthy elderly individuals, Alzheimer's disease (AD) patients, and surgical patients (Non-POD, POD) were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS / MS). Serum / plasma samples from AD mouse models and SICI mouse models were also used for validation. Results showed that serum LyPC (16:0) levels in healthy elderly individuals were significantly lower than those in healthy young adults (logFC=-0.67, adjusted-P=2.73E-06); serum LyPC (16:0) levels in AD patients were significantly lower than those in healthy elderly individuals; postoperative serum LyPC (16:0) levels in surgical patients were significantly lower than preoperative levels, and both preoperative and postoperative serum LyPC (16:0) levels in POD patients were significantly lower than those in Non-POD patients; serum / plasma LyPC (16:0) levels in AD mice and SICI mice were significantly lower than those in the corresponding normal control groups. Meanwhile, correlation analysis confirmed that LyPC (16:0) level was negatively correlated with age and significantly positively correlated with MMSE and MoCA cognitive function scores, suggesting that the absence of LyPC (16:0) is closely related to the occurrence and development of AD and SICI.

[0051] 2. The therapeutic effect of exogenous LyPC (16:0) supplementation on AD

[0052] Using 5-month-old 5×FAD mice as an AD model, 20mM LyPC (16:0) solution (200μL / time, twice a week for 6 consecutive weeks) was injected via the tail vein. The results showed that LyPC (16:0) significantly improved the spatial learning and memory abilities of AD mice (significantly increased number of platform crossings and swimming distance in the target quadrant in the Morris water maze test), new object recognition ability, and hippocampus-dependent fear memory ability; it restored the excitability of hippocampal neurons in AD mice, increased the frequency of action potential firing, increased the frequency and amplitude of microexcitatory postsynaptic currents, and improved synaptic transmission function; it significantly inhibited the overactivation of microglia and astrocytes in the hippocampus of AD mice, reduced the number of plaque-associated glial cells, reduced the levels of inflammatory factors such as TNF, IL-1β, and IL-6 in hippocampal tissue, increased dendritic spine density, and alleviated AD-related pathological damage; and it had no significant effect on Aβ plaque deposition, suggesting that it exerts its therapeutic effect by targeting neuroinflammation and synaptic function.

[0053] 3. The therapeutic and preventive effects of exogenous LyPC (16:0) supplementation on SICI

[0054] Using 18-month-old C57BL / 6J mice as the research subjects, a SICI model was established by laparotomy. One week before surgery, 20mM LyPC (16:0) solution (200μL / time, twice a week, continued for 6 weeks after surgery) was injected into the tail vein. The results showed that LyPC (16:0) could significantly improve the spatial learning and memory ability of SICI mice (shorter time to find the escape pod in the Barnes maze test and increased time spent in the target quadrant) and fear memory ability; it could restore the excitability and synaptic transmission function of hippocampal neurons in SICI mice; it could significantly inhibit the activation of glial cells in the hippocampus of SICI mice, reduce the level of neuroinflammation, increase the density of dendritic spines, and reverse SICI-related pathological damage, confirming that LyPC (16:0) has significant preventive and therapeutic effects on SICI.

[0055] 4. The preventive effect of diet / oral supplementation with LyPC (16:0) on AD.

[0056] Using 12-month-old C57BL / 6J mice as a high-risk model of age-related cognitive decline, LyPC (16:0) (5 mg / kg body weight) was administered by gavage daily for 12 months. The results showed that long-term oral supplementation with LyPC (16:0) could maintain normal levels of this metabolite in mouse serum, effectively prevent age-related cognitive decline, and significantly improve the learning and memory abilities of mice. At the same time, it could inhibit age-related hippocampal neuroinflammation, maintain dendritic spine density, protect synaptic structural integrity, and reduce the risk of AD in the elderly.

[0057] 5. The route of supplementation and formulation of LyPC (16:0)

[0058] Depending on the application scenario, multiple routes of LyPC (16:0) supplementation are provided: Injection (tail vein, intraperitoneal, subcutaneous injection) is suitable for clinical treatment, formulated as an injection, using physiological saline or physiological buffer as a solvent, with LyPC (16:0) concentration of 1μg / ml~60μg / ml. The preferred dosing regimen is 20mM / time, 200μL / time, twice a week, for no less than 6 weeks. Dietary / oral routes are suitable for daily prevention, formulated as oral preparations, capsules, tablets, functional foods, etc., with a daily supplement dose of 0.1mg~10mg / kg body weight. It can be prepared by extracting LyPC (16:0) from natural raw materials or directly adding purified LyPC, supplemented with food / pharmaceutical excipients such as maltodextrin and magnesium stearate.

[0059] The core of this invention is to protect the application of LyPC (16:0) in the preparation of drugs, foods and health products for the treatment and prevention of Alzheimer's disease (AD) and SICI. By supplementing the body with exogenous LyPC (16:0), the deficiency of LyPC (16:0) can be compensated, thereby achieving the treatment and prevention of AD and SICI.

[0060] The specific experimental steps used in this invention are as follows:

[0061] The lysophosphatidylcholine (16:0) (LyPC (16:0)) used in this invention is 1-palmitoyl-sn-glycerol-3-phosphocholine, CAS No.: 17364-16-8, and can be purchased from common reagent suppliers such as Aladdin Reagent Co., Ltd. All animals used in the experiments complied with the experimental protocol approved by the Animal Ethics Committee of Tongji University (Approval No.: TJBH00921101), and the collection of clinical samples was approved by the Ethics Committee of Shanghai Fourth People's Hospital affiliated to Tongji University (Approval No.: 2024177-001). All subjects signed written informed consent forms.

[0062] Example 1: Serum LyPC (16:0) levels decreased in patients with AD and SICI (POD).

[0063] 1. Clinical Sample Collection

[0064] A total of 48 clinical serum samples were collected and divided into 4 groups: healthy young adults (22-33 years old, n=15), healthy elderly adults (61-81 years old, n=15), Alzheimer's disease (AD) patients (n=6), and surgical patients (femoral fracture surgery, n=12, including 6 non-POD patients and 6 POD patients). Fasting serum samples were collected from surgical patients 1 hour before and 1 hour after surgery, while fasting serum samples were collected from the remaining participants once. All samples were centrifuged after standing at room temperature for 30-60 minutes. Clinical data such as age, sex, MMSE score, and MoCA score were also collected from all subjects.

[0065] 2. Detection Method

[0066] Serum metabolites were screened using non-targeted LC-MS / MS, and the content of LyPC (16:0) in serum was quantitatively detected using targeted LC-MS / MS. Figure 1 A) Spearman correlation analysis was used to assess the correlation between LyPC (16:0) level and age, MMSE score, and MoCA score.

[0067] 3. Experimental Results

[0068] 3.1 Comparison between healthy young adults and the elderly: Serum LyPC (16:0) levels in the healthy elderly population were significantly lower than those in the healthy young population (logFC=-0.67, adjusted-P=2.73E-06). Figure 1 The expression difference of B~F) and LyPC (16:0) between the two groups is shown in the figure, with the most significant difference identified as the molecule. Figure 1 G~ Figure 1 LyPC (16:0) levels were negatively correlated with age (P<0.05) and positively correlated with MMSE and MoCA scores (P<0.05). Figure 1 K~ Figure 1 N).

[0069] 3.2 Comparison between AD patients and healthy elderly individuals: Serum LyPC (16:0) levels in AD patients were significantly lower than those in healthy elderly individuals (P<0.01), and were positively correlated with MMSE and MoCA scores (P<0.05). Figure 2 A~ Figure 2 F).

[0070] 3.3 Comparison of surgical patients: Postoperative serum LyPC (16:0) levels in both POD and Non-POD patients were significantly lower than preoperative levels (P<0.05); regardless of preoperative or postoperative levels, serum LyPC (16:0) levels in POD patients were significantly lower than in Non-POD patients (P<0.05); the change in LyPC (16:0) levels before and after surgery was significantly greater in POD patients than in Non-POD patients (P<0.05). Figure 2 G~ Figure 2 K).

[0071] The above clinical sample results confirm that decreased serum LyPC (16:0) levels are closely related to the occurrence and development of AD and SICI (POD), and are an important biomarker of cognitive impairment, providing a solid clinical basis for the supplementary treatment of LyPC (16:0).

[0072] Example 2: Therapeutic effect of intravenous injection of LyPC (16:0) on AD model mice

[0073] 1. Laboratory animals and grouping

[0074] Five-month-old male 5×FAD mice (AD model) and wild-type (WT) mice of the same strain were randomly divided into three groups: WT control group, AD model group, and AD-LyPC (16:0) treatment group, with 10 mice in each group. All mice were housed in a standard animal room environment with a 12-hour light-dark cycle, a temperature of 24±1℃, and a relative humidity of 55±5%, and were allowed free access to food and water.

[0075] 2. LyPC (16:0) Formulation Preparation and Administration

[0076] LyPC (16:0) was dissolved in physiological buffer to prepare a 20 mM LyPC (16:0) solution, which was stored at 4°C for later use. Mice in the AD-LyPC (16:0) treatment group were administered 200 μL of the above solution via tail vein injection twice a week for 6 consecutive weeks; mice in the WT control group and AD model group were administered an equal volume of physiological saline via tail vein injection, with the same frequency and duration of administration as the treatment group. Figure 3 A).

[0077] 3. Detection Indicators and Methods

[0078] After drug administration, behavioral tests, electrophysiological tests, and pathological examinations were performed sequentially to comprehensively evaluate the effects of LyPC (16:0) on improving cognitive function and pathological damage in AD mice.

[0079] 3.1 Behavioral tests: The mice's motor ability was tested through the open field test and rotarod test; spatial learning and memory ability was tested through the Morris water maze test; recognition and memory ability was tested through the new object recognition test; and hippocampus-dependent fear memory ability was tested through the conditioned fear memory test. All behavioral data were automatically collected and analyzed using professional software (Ethovision XT, ANY-maze).

[0080] 3.2 Electrophysiological testing: 300 μm sagittal slices were prepared from mouse hippocampal tissue. Patch-clamp technique (Axon 700B amplifier) ​​was used to record the firing frequency of action potentials (AP) and the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSC) in hippocampal pyramidal cells to assess neuronal excitability and synaptic transmission function.

[0081] 3.3 Pathological examination: MX04 fluorescence staining was used to detect Aβ plaques in the CA1 and DG regions of the hippocampus; IBA1 and GFAP immunofluorescence staining was used to detect the activation levels of microglia and astrocytes and the number of plaque-associated glial cells; Golgi staining was used to detect the density of dendritic spines in the hippocampus; and enzyme-linked immunosorbent assay (ELISA) was used to detect the protein expression levels of inflammatory factors TNF, IL-1β, and IL-6 in the hippocampal tissue.

[0082] 4. Experimental Results

[0083] 4.1 Motor Ability: Open field and rotarod tests showed no significant differences in movement trajectory, average movement speed, and rotarod dwell time among the WT control group, AD model group, and AD-LyPC(16:0) treatment group (P>0.05), indicating that LyPC(16:0) had no effect on the motor ability of mice, thus ruling out the interference of motor ability on cognitive function testing. Figure 3 B, C).

[0084] 4.2 Cognitive Function: In the Morris water maze test, the number of platform crossings and the swimming distance in the target quadrant were significantly higher in the AD-LyPC (16:0) treatment group than in the AD model group (P<0.01), but there was no significant difference compared with the WT control group. Figure 3 D~ Figure 3 G); In the novel object recognition experiment, the novel object exploration time and recognition index of mice in the treatment group were significantly higher than those in the AD model group (P<0.05). Figure 3 H); In the conditioned fear memory experiment, the freezing time of mice in the treatment group was significantly longer than that in the AD model group (P<0.05) Figure 3 I). The above results confirm that LyPC (16:0) can significantly improve spatial learning, memory, and recognition memory abilities in AD model mice.

[0085] 4.3 Electrophysiological Results: Under 50 pA current injection, the firing frequency of action potentials in hippocampal pyramidal cells of mice in the AD-LyPC (16:0) treatment group was significantly higher than that in the AD model group (P<0.05), but there was no significant difference compared with the WT control group. Figure 3 J~3K); The frequency and amplitude of mEPSC in hippocampal neurons of the treatment group mice were significantly higher than those of the AD model group (P<0.05). This confirms that LyPC(16:0) can effectively restore the excitability and synaptic transmission function of hippocampal neurons in AD model mice (J~3K); Figure 3 L~3N).

[0086] 4.4 Pathological Results: MX04 staining showed no significant difference in fluorescence intensity of Aβ plaques in the CA1 and DG regions of the hippocampus between the AD-LyPC (16:0) treatment group and the AD model group (P>0.05). Figure 4 Immunofluorescence staining with IBA1 and GFAP showed that the number of microglia, astrocytes, and plaque-associated glial cells in the hippocampus of mice in the treatment group was significantly lower than that in the AD model group (P<0.01). Figure 4 E-4P); Golgi staining showed that the density of dendritic spines in the hippocampus of mice in the treatment group was significantly higher than that in the AD model group (P<0.01). Figure 4 T); ELISA detection showed that the protein levels of TNF, IL-1β, and IL-6 in the hippocampus of mice in the treatment group were significantly lower than those in the AD model group (P<0.05). Figure 4 Q-4S). It was confirmed that LyPC (16:0) significantly reduced glial cell activation, neuroinflammation, and dendritic spine density in AD model mice, without affecting Aβ plaque deposition.

[0087] Example 3: Preventive and therapeutic effects of intravenous injection of LyPC (16:0) on SICI model mice.

[0088] 1. Laboratory animals and grouping

[0089] Eighteen-month-old male C57BL / 6J mice were randomly divided into a blank control group, a SICI model group, and a SICI-LyPC (16:0) treatment group, with six mice in each group. The mice were kept in the same environment as in Example 1.

[0090] 2. SICI Model Construction

[0091] A SICI model was established using classic laparotomy: Mice were anesthetized with 2% isoflurane and maintained anesthesia with 1.4-2% isoflurane. The mice were placed on a temperature-controlled operating table to maintain a core body temperature of 37.5°C. A midline abdominal incision (approximately 1 cm) was made, and a 5 cm segment of small intestine was gently removed from the abdominal cavity. The intestine was wrapped with sterile saline-moistened gauze and gently manipulated for 10 minutes, simulating a clinical exploratory laparotomy. Subsequently, the muscles and skin were sutured layer by layer. Postoperatively, EMLA cream (2.5% lidocaine + 2.5% prilocaine) was applied for analgesia. The entire procedure was performed under aseptic conditions, and the surgery lasted approximately 30 minutes. The control group mice received only anesthesia without surgical intervention.

[0092] 3. LyPC (16:0) Formulation Preparation and Administration

[0093] The formulation preparation was the same as in Example 1. Mice in the SICI-LyPC (16:0) treatment group were injected with 200 μL of 20 mM LyPC (16:0) solution via the tail vein starting one week before surgery, twice a week, and continued to be administered for 6 weeks after surgery; mice in the blank control group and SICI model group were injected with an equal volume of physiological saline via the tail vein, and the frequency and cycle of administration were the same as those in the treatment group.

[0094] 4. Detection Indicators and Methods

[0095] After drug administration, behavioral tests, electrophysiological tests, and pathological tests were performed, using the same methods as in Example 1. The behavioral tests included the open field test, the conditioned fear memory test, and the Barnes maze test to assess the mice's motor ability and cognitive function. The pathological tests focused on detecting hippocampal glial cell activation, neuroinflammation, and dendritic spine density.

[0096] 5. Experimental Results

[0097] 5.1 Motor ability: Open field test results showed no significant difference in average movement speed and dwell time in the central region among the three groups of mice (P>0.05), suggesting that LyPC (16:0) had no effect on the motor ability of SICI model mice. Figure 5 BD).

[0098] 5.2 Cognitive Function: In the conditioned fear memory experiment, the freeze time of mice in the SICI-LyPC (16:0) treatment group was significantly longer than that in the SICI model group (P<0.05). Figure 5 E); In the Barnes maze test, the treatment group mice found the escape pod in a significantly shorter time than the SICI model group (P<0.01), and the proportion of time spent in the target quadrant was significantly higher in the treatment group than in the SICI model group (P<0.05). Figure 5F~5H). It was confirmed that LyPC (16:0) has a significant effect on improving cognitive function in SICI model mice, and that preoperative prophylactic supplementation can effectively reduce cognitive impairment caused by surgical trauma.

[0099] 5.3 Electrophysiological Results: Under 50 pA current injection, the firing frequency of action potentials in hippocampal pyramidal cells of mice in the SICI-LyPC (16:0) treatment group was significantly higher than that in the SICI model group (P<0.05). Figure 5 I~5J); The frequency and amplitude of mEPSCs in the hippocampal neurons of the treatment group were significantly higher than those in the SICI model group (P<0.05) Figure 5 K~5M). It was confirmed that LyPC (16:0) can restore the excitability and synaptic transmission function of hippocampal neurons in SICI model mice.

[0100] 5.4 Pathological Results: IBA1 and GFAP immunofluorescence staining showed that the number of microglia and astrocytes in the CA1 and DG regions of the hippocampus in the SICI-LyPC (16:0) treatment group was significantly lower than that in the SICI model group (P<0.01). Figure 6 A~6K); Golgi staining showed that the density of dendritic spines in the hippocampus of mice in the treatment group was significantly higher than that in the SICI model group (P<0.01). Figure 6 O); ELISA detection showed that the protein levels of TNF, IL-1β, and IL-6 in the hippocampus of mice in the treatment group were significantly lower than those in the SICI model group (P<0.05). Figure 6 LN). It was confirmed that LyPC (16:0) could significantly reduce glial cell activation, neuroinflammation, and dendritic spine density in SICI model mice.

[0101] In summary, the lysophosphatidylcholine (16:0) (LyPC (16:0)) described in this invention can be prepared on a large scale through chemical synthesis, extraction from natural animal and plant raw materials, etc., with a wide range of raw material sources and controllable production costs.

[0102] Compositions such as injections, oral formulations, capsules, tablets, and functional foods prepared based on LyPC (16:0) are all mature processes in the existing pharmaceutical and food processing fields. They can be industrialized using conventional pharmaceutical and food processing equipment without the need for additional specialized equipment.

[0103] The LyPC (16:0) and related compositions of the present invention can be widely used in many fields such as hospital clinical practice (treatment of AD and SICI patients), health care products industry (AD prevention in the elderly population and SICI prevention in high-risk surgical populations), and pharmaceutical industry (research and development and production of AD and SICI treatment drugs). There is a large market demand and broad application prospects.

[0104] In summary, this invention has significant industrial applicability and market value, and can be industrialized and promoted to generate good economic and social benefits.

[0105] The undescribed parts of this invention are the same as or implemented using existing technology. The applicant declares that this invention is illustrated through the above specific embodiments, but the invention is not limited to the above detailed methods, i.e., it does not mean that the invention must rely on the above detailed methods to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the product of this invention, additions of auxiliary components, and selection of specific methods all fall within the protection and disclosure scope of this invention.

Claims

1. The use of lysophosphatidylcholine (16:0) in the preparation of a therapeutic drug for treating cognitive impairment, characterized in that, The cognitive impairment diseases mentioned include Alzheimer's disease and postoperative cognitive impairment.

2. The use of lysophosphatidylcholine (16:0) in the preparation of foods or health products for the prevention of cognitive impairment, characterized in that, The cognitive impairment diseases mentioned include Alzheimer's disease and postoperative cognitive impairment.

3. The application according to claim 1, characterized in that, The drug uses lysophosphatidylcholine (16:0) as the sole active ingredient or in combination with other active ingredients.

4. The application according to claim 1, characterized in that, The formulation of the drug is selected from injectable or oral formulations.

5. The application according to claim 4, characterized in that, The concentration of lysophosphatidylcholine (16:0) in the injection is 1 μg / ml to 60 μg / ml.

6. The application according to claim 2, characterized in that, The daily supplemental dose of LyPC (16:0) in the food or health product is 0.1 mg to 10 mg / kg body weight.

7. The application according to claim 1 or 2, characterized in that, Lysophosphatidylcholine (16:0) is selected from natural raw materials rich in LyPC (16:0) or isolated and purified LyPC (16:0).

8. A composition for treating or preventing Alzheimer's disease or postoperative cognitive impairment, characterized in that, LyPC (16:0) is the sole or main active ingredient, supplemented with pharmaceutically acceptable carriers or food-acceptable excipients.

9. Application of reagents for detecting lysophosphatidylcholine (16:0) levels in the preparation of diagnostic or prognostic kits for Alzheimer's disease or postoperative cognitive impairment.

10. A diagnostic or prognostic kit for Alzheimer's disease or postoperative cognitive impairment, characterized in that, It contains a reagent for detecting the level of lysophosphatidylcholine (16:0).

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