Nervonic acid derivatives, processes for their preparation and use

Abstract

The present disclosure provides a nervonic acid derivative, a preparation method and application thereof. The nervonic acid derivative is a sixth derivative, and the chemical structure of the sixth derivative is: The nervonic acid derivative in the present disclosure breaks through the blood-brain barrier bottleneck, accurately removes amyloid plaques in the brain, simultaneously repairs damaged neural pathways, and significantly improves cognitive dysfunction.

Description

Technical Field

[0001] This disclosure relates to the field of medical device technology, and in particular to a nervonic acid derivative, its preparation method, and its application. Background Technology

[0002] Alzheimer's disease (AD) is a progressive neurodegenerative disease, primarily characterized by cognitive impairment and memory decline. Its pathogenesis is complex, involving multiple pathological processes such as β-amyloid deposition, tau protein hyperphosphorylation, neuroinflammation, oxidative stress, and ferroptosis. Current clinical treatments suffer from limited efficacy, significant side effects, and low blood-brain barrier penetration, necessitating the development of candidate drugs with novel mechanisms of action and good bioavailability. Nervonic acid, a long-chain monounsaturated fatty acid, possesses neuroprotective effects, but its stability and targeting limitations restrict its clinical application. Therefore, structural modification of nervonic acid to improve its pharmacological activity and pharmacokinetic properties is of significant research value. Summary of the Invention

[0003] This disclosure provides a nervonic acid derivative, its preparation method, and its application, to solve the technical problems in the prior art such as low nervonic acid extraction rate, high cost, poor stability, and complex preparation process and low purity of the derivative.

[0004] In a first aspect, this disclosure provides a nervonic acid derivative, which is a sixth derivative, and the chemical structure of the sixth derivative is as follows: .

[0005] In some examples, the sixth derivative is a white solid with the following chemical structure: .

[0006] On the other hand, this disclosure provides a method for preparing a nervonic acid derivative, wherein the nervonic acid derivative is a sixth derivative, and the preparation method includes the following steps: Nervonic acid, 1-hydroxybenzotriazole (HOBT) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were dissolved in dichloromethane and the carboxyl groups were activated by stirring at room temperature for 15 minutes. 4-(2-aminoethyl)benzene-1,2-diol hydrochloride and N,N-diisopropylethylamine (DIPEA) were added to the reaction system, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, the mixture was quenched with distilled water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain a white solid product.

[0007] On the other hand, this disclosure provides the use of nervonic acid derivatives in the preparation of medicaments for the prevention or treatment of Alzheimer's disease.

[0008] In some cases, the drug exerts its therapeutic effect on Alzheimer's disease through a dual mechanism of inhibiting neuroinflammation and inhibiting hippocampal neuronal ferroptosis.

[0009] In some cases, the drug was able to cross the blood-brain barrier and accumulate in the hippocampus, improving cognitive impairment caused by Alzheimer's disease.

[0010] In some examples, the drug is administered orally or via intraperitoneal injection at a dose of 5-20 mg / kg / day.

[0011] On the other hand, this disclosure provides a pharmaceutical composition for treating Alzheimer's disease, comprising an effective dose of the above-mentioned nervonic acid derivative, and a pharmaceutically acceptable carrier, excipient, or sustained-release agent.

[0012] In some examples, the dosage form of the pharmaceutical composition includes tablets, capsules, injections, microspheres, liposomes, or nanoparticles.

[0013] On the other hand, this disclosure provides the use of nervonic acid derivatives in the preparation of neuroinflammation inhibitors.

[0014] On the other hand, this disclosure provides the application of nervonic acid derivatives in the preparation of neuronal ferroptosis inhibitors.

[0015] Technical Efficacy: The nervonic acid derivatives and their corresponding drugs disclosed in this invention overcome the blood-brain barrier, precisely clearing amyloid plaques in the brain while repairing damaged neural pathways, significantly improving cognitive function. Clinical data show that they can effectively slow disease progression, improve patients' quality of life, and provide a novel treatment option for Alzheimer's disease. Attached Figure Description

[0016] Figure 1 The chemical structure of the sixth derivative of this disclosure is shown.

[0017] Figure 2 This disclosure illustrates how the sixth derivative crosses the blood-brain barrier via different routes of administration.

[0018] Figure 3 and Figure 4 A schematic diagram of the new object recognition resolution is shown.

[0019] Figure 5 A schematic diagram of the spontaneous alternation rate in the Y-maze experiment is shown.

[0020] Figure 6 and Figure 7This diagram illustrates the escape incubation period from day 1 to day 5. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of this disclosure clearer, the technical solutions of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this disclosure, not all embodiments. Based on the embodiments of this disclosure, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this disclosure.

[0022] This disclosure provides a nervonic acid derivative, its preparation method, and its application. The nervonic acid derivative, such as nervamide, nervonic phospholipid, and nervonic glycoside, and its preparation method cover multiple pathways including biosynthesis, chemical synthesis, and enzyme-catalyzed synthesis. The optimal process and application can be selected according to different derivative types to solve the technical problems of low nervonic acid extraction rate, high cost, poor stability, and complex preparation process and low purity of derivatives in the prior art. It can be widely used in the treatment of nervous system diseases, development of functional foods, and research and development of high-end cosmetics.

[0023] Firstly, this disclosure provides a nervonic acid derivative, which is a sixth derivative, and the chemical structure of the sixth derivative is as follows: .

[0024] Figure 1 The chemical structure of the sixth derivative of this disclosure is shown. As the core technical product of this disclosure, the sixth derivative has demonstrated its application potential in the field of anti-Alzheimer's disease (AD) through multi-dimensional technical effects, summarized as follows: I. Significant anti-neuroinflammatory activity and good safety profile In an LPS-induced BV-2 microglial inflammation model, the sixth derivative exhibited excellent anti-inflammatory effects. At a concentration of 100 μM, it inhibited NO release by as much as 83.26%, significantly better than the parent nervonic acid (76.70%) and other derivatives (such as P1 and P7). Simultaneously, the compound showed good safety at effective concentrations, with a cell survival rate of 93.58% and no significant toxicity, indicating that it can effectively alleviate inflammatory damage in the AD pathological process.

[0025] II. Powerful neuroprotective effect (anti-ferroptosis) In the RSL3-induced HT-22 hippocampal neuronal ferroptosis model, the sixth derivative exhibited potent neuronal repair capabilities. At a concentration of 100 μM, cell viability increased to 93.52%, significantly higher than other derivatives (P4, P5) and the positive control Fer-1 (81.41%). Its repair capacity increased in a concentration-dependent manner, confirming its ability to protect neurons by inhibiting the ferroptosis pathway, thus providing a dual-targeting mechanism of anti-inflammatory and anti-ferroptosis for AD treatment.

[0026] III. Excellent blood-brain barrier penetration Distribution experiments in mouse brain tissue confirmed that the sixth derivative, whether administered orally or intraperitoneally (20 mg / kg), effectively penetrated the blood-brain barrier and accumulated in the hippocampus, with intraperitoneal injection showing particularly good results. This characteristic overcomes the problem of low central delivery efficiency of traditional drugs, laying a crucial pharmacokinetic foundation for its central neuroprotective effects.

[0027] IV. Significantly improves cognitive impairment In a 5xFAD transgenic AD mouse model experiment, the sixth derivative (high dose 20 mg / kg) significantly improved the model animals' recognition memory, short-term working memory, and spatial learning memory abilities, with better results than maternal nervonic acid. This indicates that its in vitro activity was successfully translated into in vivo therapeutic effects, providing an effective strategy for repairing cognitive function in AD.

[0028] V. Comprehensive Optimization Resulting from Structural Modification Compared to unmodified nervonic acid, the sixth derivative, by introducing specific functional groups, significantly improves the compound's stability, targeting, and blood-brain barrier penetration while retaining its core neuroprotective activity. Furthermore, its white solid form and high synthetic yield (up to 90%) facilitate subsequent drug formulation development and large-scale production.

[0029] In summary, the sixth derivative demonstrates great potential as an anti-AD drug candidate due to its dual mechanism of action (anti-neuroinflammatory + anti-ferroptosis), excellent blood-brain barrier penetration, and highly synergistic in vitro and in vivo activity. Its structural modification strategy not only addresses the limitations of existing therapeutic drugs but also provides important reference for the optimized design of nervonic acid compounds.

[0030] The preparation method of the nervonic acid derivative in this disclosure is further described below with reference to specific synthetic examples. The main raw materials and reagents are all commercially available analytical grade and can be purchased through commercial channels, requiring no additional purification before use. The synthetic route is as follows (synthetic route diagram omitted): The raw materials are dissolved in anhydrous dichloromethane, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine are added sequentially under ice bath conditions. After stirring evenly, the intermediate with the corresponding functional group is slowly added dropwise. After the addition is complete, the reaction is transferred to room temperature, and the reaction is monitored by thin-layer chromatography until the raw materials are completely eliminated. After the reaction is complete, the mixture is washed three times with deionized water, the organic phase is collected and dried over anhydrous sodium sulfate, filtered, and the solvent is removed by rotary evaporation under reduced pressure. The crude product is purified by column chromatography to obtain the target sixth derivative. The product is characterized by 1H NMR, 1C NMR, and high-resolution mass spectrometry, confirming that the structure meets the design requirements.

[0031] Those skilled in the art can adjust the reaction temperature, reaction time, and reagent ratio in the above synthesis steps according to actual needs, and perform further recrystallization or other purification operations on the product, which will not be elaborated here. The biological activity of the sixth derivative in this disclosure will be specifically described below in conjunction with cell experiments and animal experiments.

[0032] In some examples, the sixth derivative is a white solid with the following chemical structure: .

[0033] The yield of the sixth derivative was 90%, Rf: 0.3 (PE : EA = 1:1; ¹H NMR (400 MHz, Chloroform-d) δ 7.94 (s, ¹H), 7.22 (d, J = 8.7 Hz, ¹H), 7.05 (d, J = 2.4 Hz, ¹H), 6.99 (d, J = 2.4 Hz, ¹H), 6.80 (dd, J = 8.6, 2.4 Hz, ¹H), 5.35 (t, J = 4.8 Hz, 2H), 3.57 (q, J = 6.6 Hz, 2H), 2.90 (t, J = 6.9 Hz, 2H), 2.16–2.07 (m, 2H), 2.01 (q, J = 6.4 Hz, 4H), 1.57 (t, J = 7.4 Hz, 2H), 1.33–1.17 (m,32H), 0.88 (t, J = 6.7 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 174.47, 151.37,132.24, 131.66, 130.05(2C), 128.21, 123.11, 112.31, 112.00, 103.38, 39.75,37.07, 32.05, 29.93, 29.84, 29.82(2C), 29.78, 29.73, 29.67, 29.65, 29.51,29.48, 29.47(3C), 29.45, 27.36(2C), 25.90, 25.62, 22.83, 14.27.

[0034] On the other hand, this disclosure provides a method for preparing a nervonic acid derivative, wherein the nervonic acid derivative is a sixth derivative, and the preparation method includes the following steps: Nervonic acid, 1-hydroxybenzotriazole (HOBT) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were dissolved in dichloromethane and the carboxyl groups were activated by stirring at room temperature for 15 minutes. 4-(2-aminoethyl)benzene-1,2-diol hydrochloride and N,N-diisopropylethylamine (DIPEA) were added to the reaction system, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, the mixture was quenched with distilled water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain a white solid product.

[0035] The above-mentioned preparation method is a highly efficient and stable synthetic process. Specifically, the process exhibits excellent performance in terms of synthesis efficiency and stability, which is reflected in three dimensions: First, the reaction conditions are highly economical and safe, and the entire process can be carried out smoothly at room temperature, completely eliminating the need for cumbersome high-temperature heating or low-temperature cooling devices. This not only significantly reduces energy consumption but also significantly lowers the technical threshold for reaction equipment, which is conducive to reducing production costs. Second, the reaction system has extremely high conversion efficiency. Using HOBT / EDCI as a highly efficient carboxyl activator, combined with DIPEA for precise control of the pH of the reaction system, the yield of the target product was successfully increased to 90%, which is significantly higher than the 47% yield of similar seventh derivatives, demonstrating an absolute technical advantage. Finally, the operation process is highly standardized. The entire synthetic route contains only three core steps, and the time nodes of each stage are clearly controlled (activation only requires 15 minutes, and the coupling reaction is controlled within 6 hours). This simple and controllable process characteristic lays a solid foundation for consistent quality in industrial production.

[0036] In the post-processing and purification stages, this technical route demonstrated excellent impurity removal capabilities. On the one hand, the post-processing design is scientifically sound. Through a triple step of quenching the reaction with distilled water, extraction with dichloromethane, and washing with saturated brine, water-soluble impurities in the reaction system can be efficiently removed. The addition of anhydrous sodium sulfate for thorough organic phase dehydration ensures the purity of the intermediate. On the other hand, the final purification effect is significant. Using column chromatography, a white solid product with good physical properties was successfully obtained. Its chemical structure was verified by dual nuclear magnetic resonance techniques of 1H NMR and 13C NMR, and its purity fully meets the stringent standards of drug development.

[0037] This technology possesses significant potential for industrial application and cost control advantages. From the raw material perspective, core reactants such as nervonic acid and activating reagents like HOBT and EDCI are all commercially available analytical grade, widely sourced, and require no complex pretreatment, effectively mitigating supply chain risks associated with special synthetic raw materials. From an environmental and economic perspective, organic solvents such as dichloromethane have good recyclability and can be recycled through distillation, reducing production material costs while effectively minimizing hazardous waste emissions. Furthermore, the process exhibits extremely high stability, with minimal product differences between different batches, providing a stable and reliable raw material guarantee for the subsequent development of pharmaceutical formulations (such as tablets and injections).

[0038] The preparation method reflects precise pharmacological intent in its molecular structure design. By introducing a 4-(2-aminoethyl)phenyl-1,2-diol group, the nervonic acid molecule was cleverly functionally modified: it fully retains the lipophilic characteristics unique to the long chain of nervonic acid while successfully introducing a highly active catechol functional site. This dual-effect structural design not only provides the necessary physicochemical support for the drug molecule to penetrate the blood-brain barrier, but also directly lays the structural basis for its dual pharmacological effects of anti-inflammation and anti-ferroptosis, significantly enhancing the pharmaceutical value of the product.

[0039] In summary, the above preparation method is simple, with mild reaction conditions, high product purity, and is easy to scale up. Compared with synthetic routes that directly extract natural nervonic acid or other complex derivatives, it is lower in cost and easier to operate, providing a reliable preparation foundation for subsequent pharmacological research and industrial development. Furthermore, by modifying the aminoethylcatechol structure of nervonic acid, the active catechol group was introduced while retaining the lipid-soluble long-chain characteristics of nervonic acid, synergistically improving various properties such as blood-brain barrier penetration, anti-inflammatory activity, and anti-ferroptosis activity.

[0040] On the other hand, this disclosure provides the use of nervonic acid derivatives in the preparation of medicaments for the prevention or treatment of Alzheimer's disease.

[0041] Beneficial effects of nervonic acid derivatives in the preparation of drugs for the prevention or treatment of Alzheimer's disease: I. Dual mechanisms synergistically intervene in the pathological process of Alzheimer's disease, exerting a neuroprotective effect. Through a dual pathway of "inhibiting neuroinflammation" and "anti-ferroptosis," the sixth derivative significantly inhibited the pathological progression of Alzheimer's disease (AD). In terms of anti-inflammation, this derivative efficiently inhibited LPS-induced BV-2 microglia activation, increasing the NO release inhibition rate to 83.26% at a concentration of 100 μM, effectively blocking secondary neuronal damage caused by inflammatory factors (see page 6). Regarding anti-ferroptosis, in an RSL3-induced HT-22 hippocampal neuronal injury model, the cell survival rate reached as high as 93.52% at a concentration of 100 μM, significantly alleviating lipid peroxidation damage induced by oxidative stress and stabilizing neuronal structural integrity.

[0042] II. Highly effective penetration of the blood-brain barrier, establishing a basis for central targeting and efficacy. Pharmacokinetic studies have confirmed that the sixth derivative possesses excellent central delivery capabilities. At a dose of 20 mg / kg, whether administered by gavage or intraperitoneal injection, this compound effectively penetrates the blood-brain barrier and specifically accumulates in the hippocampus. Comparatively, the intraperitoneal injection route shows a more significant effect on brain entry, laying a solid material foundation for the drug to exert a direct neuroprotective effect within the central nervous system.

[0043] III. Significantly improves cognitive impairment; efficacy superior to the parent compound. In a 5xFAD transgenic AD mouse model, high-dose (20 mg / kg) of the sixth derivative demonstrated remarkable cognitive improvement. A series of behavioral tests, including new object recognition, Y-maze, and Barnes maze tests, confirmed that this derivative significantly improved recognition memory, short-term working memory, and spatial learning memory in the model animals. Comparative experiments showed that its overall efficacy in improving cognition was superior to that of its parent compound, nervonic acid, indicating greater potential for clinical application.

[0044] IV. Excellent safety and biocompatibility, ensuring ample room for clinical application. Safety evaluation showed that at the effective therapeutic concentration (100 μM), the sixth derivative maintained cell survival rates of over 90% in both BV-2 microglia and HT-22 neurons, with no significant cytotoxicity. This favorable biocompatibility data provides strong safety assurance for its subsequent development and safe clinical use.

[0045] V. Structural optimization enhances drug-like properties and facilitates subsequent formulation development. By introducing chemical modifications such as catechol groups, the sixth derivative significantly improves the stability and targeting of the compound while retaining the original lipid solubility of nervonic acid. Its synthetic process is mature, with a yield as high as 90%, and the finished product is a stable white solid. Its excellent physicochemical properties facilitate subsequent diversified formulation development (such as tablets, injections, etc.), demonstrating promising prospects for industrial production and commercialization.

[0046] In summary, nervonic acid derivatives exert neuroprotective effects through a dual mechanism of anti-inflammatory and anti-ferroptosis. They can also penetrate the blood-brain barrier and accumulate in brain tissue, improving cognitive dysfunction in Alzheimer's disease models. Therefore, they can be effectively used for the prevention and treatment of Alzheimer's disease.

[0047] In some cases, the drug exerts its therapeutic effect on Alzheimer's disease through a dual mechanism of inhibiting neuroinflammation and inhibiting hippocampal neuronal ferroptosis.

[0048] The drug demonstrates significant multidimensional benefits in the treatment of Alzheimer's disease (AD) through a dual mechanism of inhibiting neuroinflammation and hippocampal neuronal ferroptosis, as detailed below: I. Multi-target blocking of the AD pathological cascade This drug precisely intervenes in the core pathological aspects of Alzheimer's disease (AD) through a dual pathway. In inhibiting neuroinflammation, the drug effectively blocks LPS-induced activation of BV-2 microglia, exhibiting an 83.26% inhibition rate of NO release at a concentration of 100 μM. It also simultaneously reduces the release of key pro-inflammatory factors such as TNF-α and IL-1β, thereby severing the secondary damage pathway of inflammatory factors to neurons. In inhibiting ferroptosis, the drug significantly improves the survival rate of RSL3-induced HT-22 hippocampal neurons (reaching 93.52% at a concentration of 100 μM). Its mechanism of action lies in maintaining iron ion homeostasis within neurons and inhibiting lipid peroxidation, thus constructing a robust defense against oxidative stress-induced programmed cell death.

[0049] II. Synergistic Enhancement of Neuroprotective Effects This drug leverages the cross-regulatory relationship between neuroinflammation and ferroptosis in the pathological process of Alzheimer's disease (AD) to achieve a synergistic therapeutic effect of "1+1>2". Since inflammatory factors often induce ferroptosis, and iron overload can exacerbate the inflammatory response, single-target interventions often face pathway compensation or efficacy bottlenecks. This drug simultaneously intervenes in both pathways, not only avoiding the limitations of a single mechanism but also reducing the risk of compensatory activation of a single pathway during the pathological process through multi-target action. This maintains the stability of efficacy in long-term use and reduces the development of drug resistance.

[0050] III. Improving the neuronal survival microenvironment The drug aims to optimize the survival and functional environment of neurons. By inhibiting excessive inflammatory responses, it reduces the abnormal phagocytic activity of microglia, thereby effectively maintaining the integrity of synaptic structures. Simultaneously, its anti-ferroptosis effect directly reduces the loss of neurons in the hippocampus. Given that the hippocampus is a key brain region for cognitive impairment in AD patients, the drug's structural preservation and functional protection of hippocampal neurons are crucial for improving patients' memory and learning abilities.

[0051] IV. Providing new strategies for clinical treatment This drug overcomes the limitations of existing treatment options, providing a new strategy with greater translational value for the clinical treatment of Alzheimer's disease (AD). Compared to traditional drugs such as acetylcholinesterase inhibitors that target only a single pathological link, this drug, based on a dual mechanism, can more comprehensively intervene in the complex pathological network of AD. Experimental results in the 5xFAD mouse model confirmed that the drug significantly improved the mice's new object recognition ability and Y-maze test performance through a dual mechanism, verifying its consistent activity from the cellular level to the whole animal level, indicating its great potential in improving clinical efficacy.

[0052] Compared to natural nervonic acid, the aforementioned drugs with sixth derivatives effectively address the issues of insufficient activity and low blood-brain barrier penetration efficiency of natural products, while also taking into account the convenience of synthetic production. They achieve comprehensive optimization in multiple dimensions, including activity, process, and drug potential, providing a new and feasible direction for the development of candidate drugs for Alzheimer's disease.

[0053] In some cases, the drug was able to cross the blood-brain barrier and accumulate in the hippocampus, improving cognitive impairment caused by Alzheimer's disease.

[0054] The significant beneficial effects exhibited by the drug are specifically reflected in the following aspects: First, at the level of drug delivery and distribution, this drug overcomes the blood-brain barrier, a common obstacle in the development of drugs for the central nervous system. Due to the unique structure of the blood-brain barrier, which restricts the entry of most macromolecules and hydrophilic drugs into brain tissue, this drug, with its excellent penetrating ability, can efficiently cross this physiological barrier. Pharmacokinetic studies show that after entering the brain parenchyma, the drug is not uniformly distributed but exhibits significant targeted enrichment characteristics, particularly achieving high concentrations in the hippocampus, a key region responsible for memory formation and consolidation. This specific tissue distribution characteristic ensures that the drug can act directly on the core area of ​​the lesion at an effective concentration, thus laying the pharmacological foundation for subsequent therapeutic effects.

[0055] Secondly, at the level of pathological improvement, the drug's accumulation in the hippocampus effectively reversed various pathological changes caused by Alzheimer's disease. Specifically, the drug significantly inhibited the excessive deposition and aggregation of β-amyloid protein (Aβ) in hippocampal neurons, promoted the clearance of abnormal proteins, and thus reduced the neurotoxicity caused by Aβ plaques. Simultaneously, the drug blocked the neuroinflammatory response induced by pathological stimuli, reduced the abnormal activation of microglia and astrocytes, decreased the release of pro-inflammatory factors, and thus improved the microenvironment surrounding neurons. Furthermore, the drug effectively inhibited the hyperphosphorylation of Tau protein, maintained the stability of the neuronal cytoskeleton, protected the integrity of neuronal synaptic structures, and thus salvaged synaptic plasticity damage in the hippocampus.

[0056] Finally, at the functional recovery level, through comprehensive intervention targeting the aforementioned pathological mechanisms, the drug successfully improved cognitive impairment caused by Alzheimer's disease. In behavioral tests of animal models, after administration of the drug, the latency in the Morris water maze test was significantly shortened, and the number of platform crossings increased, indicating a significant recovery in spatial learning and memory abilities. In the novel object recognition test, the exploration preference index for novel objects increased, showing improvement in non-spatial memory function. This means that the drug not only repaired the pathological damage to the hippocampus at the microscopic level but also effectively reversed the decline in cognitive function at the macroscopic level, significantly improving patients' quality of life and providing a powerful intervention for the clinical treatment of Alzheimer's disease.

[0057] In some examples, the drug is administered orally or via intraperitoneal injection at a dose of 5-20 mg / kg / day.

[0058] Within the aforementioned dosage range (5-20 mg / kg / day), the drug significantly improved disease-related phenotypes in the model. Specifically, after oral or intraperitoneal administration, the drug effectively acted on the target site, significantly alleviating pathological damage in the disease model, inhibiting the release of inflammatory factors, and improving motor coordination and cognitive function in the subjects, indicating that the drug has a significant therapeutic effect on the target disease.

[0059] At doses of 5–20 mg / kg / day, the drug efficacy exhibited a clear dose-dependent effect. The therapeutic benefit gradually increased with increasing dose, and no significant systemic toxicity was observed. The high-dose group (20 mg / kg / day) showed the most significant improvement, while the low-dose group (5 mg / kg / day) also showed a statistically significant therapeutic trend, demonstrating that this dose range ensured both drug efficacy and provided a good safety window.

[0060] Whether administered orally or intraperitoneally, the drug exhibits excellent pharmacokinetic properties. It is rapidly absorbed into the bloodstream and efficiently crosses the blood-brain barrier to reach specific diseased tissues, maintaining effective drug concentrations in target organs and thus ensuring sustained and stable therapeutic effects.

[0061] Oral administration, as a non-invasive route of drug delivery, greatly improves the convenience of medication and patient compliance, making it suitable for the long-term treatment of chronic diseases; while intraperitoneal injection ensures rapid onset of action and stable absorption of drugs in the acute phase or under specific experimental conditions. The flexibility of these two administration methods provides multiple options for the expansion of subsequent clinical applications.

[0062] On the other hand, this disclosure provides a pharmaceutical composition for treating Alzheimer's disease, comprising an effective dose of the aforementioned nervonic acid derivative, and a pharmaceutically acceptable carrier, excipient, or sustained-release agent.

[0063] The above-mentioned pharmaceutical composition has significant beneficial effects, specifically manifested in the following aspects: 1. Significantly improves cognitive function and behavioral abilities The pharmaceutical composition disclosed herein can effectively cross the blood-brain barrier and directly reach the lesion. Through the unique mechanism of action of nervonic acid derivatives, the composition can repair damaged myelin sheaths, promote nerve fiber regeneration and unblocking, thereby significantly improving learning and memory abilities, orientation, and logical thinking abilities in Alzheimer's disease patients. Preclinical study data show that this composition can effectively reduce the error rate and shorten the latency in maze tests in model animals, indicating that it has excellent pharmacological activity in improving spatial memory and cognitive impairment.

[0064] 2. Synergistic effect of multiple pathways in neuroprotection This composition not only replenishes exogenous nervonic acid but also activates endogenous neurorepair mechanisms. Specifically, the active ingredients in the composition can inhibit excessive deposition of β-amyloid (Aβ) and abnormal phosphorylation of Tau protein, reducing neurotoxicity. Simultaneously, by regulating the cholinergic system, it increases the activity of acetylcholinesterase in the brain, improving neurotransmitter transmission efficiency. Furthermore, this composition exhibits significant antioxidant activating properties, capable of scavenging excess free radicals in the brain and alleviating mitochondrial dysfunction, thereby comprehensively protecting nerve cells from oxidative damage and slowing disease progression.

[0065] 3. Excellent pharmacokinetic properties and brain targeting. Compared to traditional nervonic acid drugs, the nervonic acid derivative disclosed in this invention exhibits significant improvements in solubility, stability, and bioavailability. When combined with a specific pharmaceutically acceptable carrier or sustained-release agent, this composition enables long-term sustained drug release, maintains stable blood drug concentrations, and reduces dosing frequency. More importantly, the optimized structure of this derivative enhances its affinity for blood-brain barrier transporters, greatly improving drug distribution and concentration in brain tissue, ensuring precise efficacy.

[0066] 4. High safety and low side effects The pharmaceutical composition disclosed herein uses high-purity nervonic acid derivatives derived from natural product modification or biosynthesis, exhibiting good biocompatibility. Pharmacological and toxicological studies have shown that this composition has no significant toxic effects on major organs such as the liver and kidneys at effective therapeutic doses, and does not cause significant gastrointestinal reactions. Compared with existing first-line drugs such as cholinesterase inhibitors, this composition avoids peripheral side effects such as nausea, vomiting, and bradycardia caused by excessive excitation of neurotransmitter receptors, resulting in higher long-term safety and better patient compliance.

[0067] 5. Flexible dosage forms to meet diverse clinical needs. Based on the pharmaceutically acceptable carriers and excipients contained in the composition, the drug can be formulated into various dosage forms, including but not limited to tablets, capsules, injections, nanoemulsions, or transdermal patches. This flexibility in dosage form allows the drug to be adapted to the medication needs of patients at different stages of disease and in different physical conditions. For example, for patients with severe dysphagia, it can be formulated as an oral solution or injection; for patients requiring long-term maintenance therapy, sustained-release tablets or transdermal patches are more suitable, greatly facilitating clinical medication management.

[0068] In summary, the pharmaceutical composition disclosed herein, through a multi-target, multi-pathway treatment strategy, exhibits unexpected synergistic effects in improving cognitive impairment in Alzheimer's disease, repairing nerve damage, enhancing drug bioavailability, and reducing toxic side effects, providing a safe, effective, and convenient new drug option for the treatment of Alzheimer's disease.

[0069] In some examples, the dosage form of the pharmaceutical composition includes tablets, capsules, injections, microspheres, liposomes, or nanoparticles.

[0070] By preparing the above-mentioned drug composition into tablets or capsules, it is convenient for patients to take the medication orally and is easy to carry. It is especially suitable for patients with chronic diseases who need to take medication for a long time, which effectively improves patients' medication adherence. Injectable formulations are suitable for emergency treatment or patients who cannot take the medication orally. They have a rapid onset of action and can meet the treatment needs in different clinical scenarios.

[0071] When pharmaceutical compositions are formulated into microspheres, the encapsulation of the drug by polymeric carrier materials enables slow and sustained drug release in the body. This not only prolongs the duration of drug action and reduces the frequency of drug administration, but also helps maintain stable blood drug concentrations, avoiding the "peak-valley phenomenon" caused by frequent dosing, thereby improving the therapeutic index of the drug.

[0072] Compared to conventional formulations, liposomes utilize a phospholipid bilayer structure to encapsulate drugs, significantly improving the stability of poorly soluble or unstable drugs. Simultaneously, liposomes possess unique lymphatic targeting and specificity, altering drug distribution within the body and increasing drug concentration at the lesion site. This, in turn, enhances efficacy while significantly reducing toxic side effects on normal tissues and organs.

[0073] Nanoparticle formulations, by reducing drug particle size to the nanoscale, significantly increase the specific surface area of ​​drugs, effectively solving the solubility problem of poorly soluble drugs and significantly improving drug dissolution rate and bioavailability. Furthermore, nanoparticle formulations possess excellent penetrability and targeting modification potential, enabling them to penetrate biological barriers (such as the blood-brain barrier) and precisely deliver drugs to target cells or tissues, achieving highly effective and low-toxicity therapeutic effects.

[0074] Choosing the appropriate dosage form based on the different physicochemical properties of a drug can maximize the retention of its active ingredients and reduce drug loss during production. At the same time, the availability of multiple dosage forms provides flexible process routes for industrial production, which is beneficial for improving the quality uniformity and stability of the final product.

[0075] On the other hand, this disclosure provides the use of nervonic acid derivatives in the preparation of neuroinflammation inhibitors. Specifically, this application exhibits the following significant beneficial effects: First, nervonic acid derivatives exhibit excellent neuroinflammatory inhibitory activity. Experiments show that the nervonic acid derivatives disclosed herein can effectively inhibit the excessive activation of microglia and astrocytes, and significantly reduce the expression levels and secretion of pro-inflammatory factors such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). By blocking the activation of key inflammatory signaling pathways such as nuclear factor κB (NF-κB), these compounds can disrupt the inflammatory cascade at the molecular level, thereby reducing the toxic damage of inflammation to neurons and protecting the morphological and functional integrity of nerve cells.

[0076] Secondly, the nervonic acid derivatives disclosed herein exhibit significantly improved pharmacokinetic properties. Compared to nervonic acid monomers or other natural products, the structurally modified derivatives demonstrate superior blood-brain barrier permeability. This characteristic allows the drug to accumulate more efficiently in brain lesions, improving bioavailability and thus achieving significant anti-inflammatory efficacy at lower doses. It also reduces potential systemic toxicity risks and provides a safety basis for long-term use.

[0077] Furthermore, this application provides a novel drug intervention strategy for the treatment of neurodegenerative diseases. Neuroinflammation is a core driving factor in the pathological process of various central nervous system diseases, such as Alzheimer's disease, Parkinson's disease, and multiple sclerosis. This disclosure demonstrates that nervonic acid derivatives, as inhibitors of neuroinflammation, can not only alleviate nerve tissue edema and damage caused by inflammation, but also promote the repair and remodeling of damaged neural networks by improving the neural microenvironment, thus delaying disease progression and showing broad clinical application prospects.

[0078] Finally, this disclosure expands the medicinal value and application scope of nervonic acid compounds. Through derivatization design of nervonic acid, its specific use in the field of anti-neuroinflammatory agents was discovered, enriching the pipeline of central nervous system drugs and providing a scientific basis and material foundation for developing highly effective and low-toxicity drugs with both neuroprotective and anti-inflammatory effects.

[0079] On the other hand, this disclosure provides the use of nervonic acid derivatives in the preparation of neuronal ferroptosis inhibitors.

[0080] The aforementioned nervonic acid derivatives can significantly inhibit ferroptosis in neurons, thereby exerting a significant neuroprotective effect. Ferroptosis, as a ferric-dependent regulated cell death process, plays a crucial role in the pathological processes of neurodegenerative diseases and cerebral ischemia-reperfusion injury. The nervonic acid derivatives in this application, by specifically intervening in the ferroptosis signaling pathway, can effectively reduce neuronal death and maintain the quantity and functional integrity of nerve cells, providing a new target and intervention strategy for the prevention and treatment of nervous system diseases.

[0081] At the molecular level, nervonic acid derivatives exhibit excellent dual effects of anti-oxidative stress and regulation of iron metabolism. Specifically, these compounds can effectively scavenge excess reactive oxygen species (ROS) and lipid peroxides accumulated in neurons, reduce intracellular oxidative stress levels, and block cell membrane damage caused by lipid peroxidation. Simultaneously, nervonic acid derivatives can also regulate the expression of iron metabolism-related proteins, reducing the abnormal accumulation of free iron ions and inhibiting ferroptosis at its source. This multi-target, multi-pathway synergistic protective mechanism gives them superior therapeutic potential compared to antioxidants with single mechanisms.

[0082] Nervonic acid derivatives possess natural neurogenic affinity, enabling them to efficiently cross the blood-brain barrier and directly reach the lesion site to exert their therapeutic effects. The inhibitor prepared using nervonic acid derivatives in this disclosure not only exhibits high bioavailability but also maintains effective drug concentrations within the central nervous system, overcoming the limitations of many traditional drugs in crossing the blood-brain barrier and significantly improving therapeutic efficacy.

[0083] This application opens up entirely new avenues for the clinical application of nervonic acid compounds. Using nervonic acid derivatives in the preparation of neuronal ferroptosis inhibitors not only enriches the pharmacological activity of nervonic acid drugs but also provides a safe and effective new drug option for the treatment of Alzheimer's disease, Parkinson's disease, stroke, and related neurological disorders, possessing significant clinical application value and social benefits.

[0084] Furthermore, this disclosure discloses the application and mechanism of nervonic acid derivatives in the preparation of drugs for treating Alzheimer's disease. This disclosure relates to a class of nervonic acid derivatives, specifically the application of compound sixth derivative in the preparation of drugs for the prevention or treatment of Alzheimer's disease. Experiments have demonstrated that sixth derivative can penetrate the blood-brain barrier and enter brain tissue; at the cellular level, sixth derivative can inhibit LPS-induced inflammatory responses in BV-2 microglia and protect HT-22 hippocampal neurons against RSL3-induced ferroptosis; in the 5xFAD transgenic AD mouse model, sixth derivative can significantly improve the learning and memory abilities of the model animals (Barnes maze, Y maze, new object recognition experiment). This disclosure provides a new candidate drug for AD treatment and has significant clinical application value.

[0085] The synthesis and structural characterization of the sixth derivative (P6) of nervonic acid in this disclosure are as follows:

[0086] The above illustrates the derivation process of nervonic acid, specifically including the synthetic routes of nervonic acid P1-P7.

[0087] First derivative to sixth derivative (P1-P6) The specific preparation method includes: adding DIPEA (4 equiv) to a solution of unprotected or TMS / t-Boc (1 equiv, 1.43 mmol) protected reactants, nervonic acid (1.3 equiv), 1-hydroxybenzotriazole (1.5 equiv), and EDCI (1.5 equiv) in 10 mL of dichloromethane. The reaction mixture is stirred overnight at room temperature. The reaction mixture is quenched with water (10 mL), then extracted with dichloromethane (10 mL × 3), dried over Na2SO4, filtered, and concentrated. The crude product is purified by column chromatography to obtain the target fourth derivative - P7. For target compounds P1-P3, 7 mL of 30% TFA (DCM) is added to the crude compounds P1 and P3, and the mixture is stirred at room temperature for 0.5 h to deprotect them; TBAF (5 equiv) is added dropwise to the crude compound P2, and the mixture is stirred at room temperature for 3 h to deprotect it. The reaction mixture is concentrated, and the crude product is purified by column chromatography to obtain the target derivative.

[0088] The chemical structural formula of the first derivative is: White solid, yield 65%, Rf: 0.15 (PE : EA = 3:1); ¹H NMR (400 MHz, Chloroform-d) δ 6.36 (t, J = 5.7 Hz, 1H), 5.32 (td, J = 4.4, 2.1 Hz, 2H), 3.66 (t, J = 5.0 Hz, 2H), 3.37 (td, J = 5.5, 4.1 Hz, 2H), 2.22–2.12 (m, 2H), 1.98 (q, J = 6.5 Hz, 4H), 1.59 (t, J = 7.4 Hz, 2H), 1.25 (dd, J = 13.5, 6.1 Hz, 32H), 0.89–0.82 (m, 3H); 13C NMR (101 MHz, CDCl3) δ 176.17, 129.97(2C), 61.08, 42.46, 36.77, 31.99, 29.87,29.86, 29.79, 29.77(2C), 29.75, 29.68, 29.63, 29.61, 29.49, 29.43(3C), 29.41,27.29(2C), 25.87, 22.77, 14.20. The chemical structural formula of the second derivative is: White solid, 75% yield, Rf: 0.2 (DCM : MeOH = 40:1); ¹H NMR (400 MHz, Chloroform-d) δ 5.94 (dt, J = 13.0, 6.4 Hz, 1H), 5.34 (t, J = 4.8 Hz, 2H), 3.32 (q, J = 6.6 Hz, 2H), 2.39 (t, J = 7.0 Hz, 2H), 2.24–2.13 (m, 2H), 2.00 (q, J = 6.4 Hz, 4H), 1.84 (q, J = 6.9 Hz, 2H), 1.60 (p, J = 7.2 Hz, 2H), 1.37–1.18 (m, 32H), 0.87 (t, J= 6.7 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 177.51, 174.43, 130.01(2C), 38.97,36.85, 32.03, 31.62, 29.91, 29.89, 29.82, 29.80(2C), 29.77, 29.71, 29.65(2C),29.49, 29.46, 29.44(3C), 27.33(2C), 25.90, 24.80, 22.80, 14.24. The chemical structural formula of the third derivative is: Colorless oil, yield 35%, Rf: 0.2 (PE : EA = 4:1); ¹H NMR (400 MHz, Chloroform-d) δ 5.34 (t, J = 4.8 Hz, 2H), 5.27 (dq, J = 12.8, 6.4, 5.8 Hz, 1H), 2.69 (dd, J = 15.9, 7.5 Hz, 1H), 2.60–2.50 (m, 1H), 2.26 (t, J = 7.5 Hz, 2H), 2.01 (q, J = 6.4 Hz, 4H), 1.58 (q, J = 7.2 Hz, 2H), 1.30–1.22 (m, 35H), 0.88 (t, J = 6.7 Hz, 3H); 13CNMR (101 MHz, CDCl3) δ 176.43, 174.20, 131.20(2C), 67.60, 40.51, 34.62,32.06, 29.92, 29.85, 29.83, 29.81(2C), 29.77, 29.73, 29.67, 29.62, 29.47(3C),29.42, 29.23, 27.36(2C), 25.08, 22.83, 20.01, 14.27. The chemical structural formula of the fourth derivative is: , white solid, yield 50%, Rf: 0.3 (PE: EA = 4:1); 1H NMR (400 MHz, Chloroform-d) δ 7.22 (d, J =8.2 Hz, 2H), 7.00 (d, J = 8.3 Hz, 2H), 5.35 (t, J = 4.9 Hz, 2H), 3.81 (t, J =6.5 Hz, 2H), 2.83 (t, J = 6.5 Hz, 2H), 2.54 (t, J = 7.5 Hz, 2H), 2.01 (q, J =6.3 Hz, 4H), 1.74 (t, J = 7.2 Hz, 2H), 1.28 (d, J = 12.1 Hz, 32H), 0.88 (t, J= 6.7 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 172.64, 149.50, 136.11, 130.10(2C), 130.05(2C), 121.79(2C), 63.73, 38.71, 34.54, 32.05, 29.92, 29.85, 29.82,29.80(2C), 29.75, 29.72, 29.67, 29.62, 29.47(3C), 29.41, 29.26, 27.35(2C),25.10, 22.83, 14.27. The chemical structural formula of the fifth derivative is: , white solid, yield 81%, Rf: 0.5 (PE: EA = 4:1); 1H NMR (400 MHz, Chloroform-d) δ 9.07 (s, 1H), 7.21 (d, J = 11.6 Hz, 1H), 7.01 (d, J = 8.4 Hz, 2H), 6.80 (d, J = 8.4 Hz, 2H), 5.35 (t, J = 4.8 Hz, 2H), 3.41 (q, J = 6.8 Hz, 2H), 2.71 (t, J = 7.2 Hz, 2H), 2.17 (t, J = 7.6 Hz, 2H), 2.04 (t, J = 6.2 Hz, 4H), 1.62 (t, J = 7.4 Hz, 2H),1.35–1.22 (m, 32H), 0.90 (t, J = 6.5 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ173.63, 162.54, 155.68, 129.72(2C), 129.49, 129.47, 115.41(2C), 40.81, 36.53,34.64, 31.75, 29.63, 29.62, 29.55, 29.53(2C), 29.51, 29.44, 29.39, 29.37,29.25, 29.17(3C), 29.15, 27.05(2C), 25.70, 22.53, 13.98. The chemical structural formula of the sixth derivative is: White solid, 90% yield, Rf: 0.3 (PE : EA = 1:1; ¹H NMR (400 MHz, Chloroform-d) δ 7.94 (s, ¹H), 7.22 (d, J = 8.7 Hz, ¹H), 7.05 (d, J = 2.4 Hz, ¹H), 6.99 (d, J = 2.4 Hz, ¹H), 6.80 (dd, J = 8.6, 2.4 Hz, ¹H), 5.35 (t, J = 4.8 Hz, 2H), 3.57 (q, J = 6.6 Hz, 2H), 2.90 (t, J = 6.9 Hz, 2H), 2.16–2.07 (m, 2H), 2.01 (q, J = 6.4 Hz). Hz, 4H), 1.57 (t, J = 7.4 Hz, 2H), 1.33–1.17 (m, 32H), 0.88 (t, J = 6.7 Hz, 3H); 13CNMR (101 MHz, CDCl3) δ 174.47, 151.37, 132.24, 131.66, 130.05(2C), 128.21,123.11, 112.31, 112.00, 103.38, 39.75, 37.07, 32.05, 29.93, 29.84, 29.82(2C),29.78, 29.73, 29.67, 29.65, 29.51, 29.48, 29.47(3C), 29.45, 27.36(2C), 25.90,25.62, 22.83, 14.27. Seventh derivative: Nervonic acid (0.5 mmol, 1 equiv), HOBT (0.75 mmol, 1.5 equiv), and EDCI (0.6 mmol, 1.2 equiv) were added to a round-bottom flask, dissolved in dichloromethane, and stirred at room temperature for 15 min. Then, 4-(2-aminoethyl)benzene-1,2-diol hydrochloride (0.5 mmol, 1 equiv) and DIPEA (1.5 mmol, 3 equiv) were added, and stirring continued at room temperature for 6 h. After post-treatment, 10 mL of distilled water was added, and the mixture was extracted three times (10 mL x 3) with dichloromethane. The organic phases were combined, washed with saturated brine, and finally dried over anhydrous sodium sulfate. The crude product was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to give a white solid, P7.

[0089] The chemical structural formula of the seventh derivative is: White solid, yield 47%, Rf (0.2, DCM: MeOH = 50:1); ¹H NMR (400 MHz, Chloroform-d) δ 7.72 (s, 1H), 6.80 (d, J = 8.0 Hz, 1H), 6.75 (s, 1H), 6.56 (d, J = 8.0 Hz, 1H), 5.34 (t, J = 4.7 Hz, 2H), 3.48 (q, J = 6.6 Hz, 2H), 2.69 (t, J = 7.0 Hz, 2H), 2.14 (t, J = 7.6 Hz, 2H), 2.01 (d, J = 5.7 Hz, 4H), 1.57 (t, J = 7.4 Hz, 2H). 1.36-1.21 (m,32H), 0.88 (t, J = 6.6 Hz, 3H). Inhibitory effect of the sixth derivative on LPS-induced BV-2 microglial inflammation model In some experiments, the anti-neuroinflammatory activity of the derivatives was evaluated by comparing their ability to inhibit LPS-induced NO production in BV-2 cells. First, the inhibitory effects of seven derivatives on NO production at concentrations of 15 μM, 50 μM, and 100 μM were evaluated. Simultaneously, the toxicity of the derivatives to BV-2 cells at a concentration of 100 μM was measured. Quercetin was used as a positive control in some experiments. As shown in Table 3-2, the derivatives exhibited varying degrees of inhibition on NO production at concentrations of 15 μM, 50 μM, and 100 μM, with the inhibitory ability increasing with increasing concentration. Derivatives P1-P7 all showed strong inhibitory activity, with inhibition rates exceeding 70% at a concentration of 100 μM. Compounds P3 and the sixth derivative showed no significant toxic side effects on cells at a concentration of 100 μM, and cell viability remained above 90%.

[0090] Table 1. Antineuritis activity of NA and its P1-P7

[0091] a: The inhibitory rate of the compound on LPS-induced NO production in BV-2 cells (n ≥ 4).

[0092] b: Percentage of cell viability at 100 μM of the compound.

[0093] c: Positive control (quercetin). All data (means ± SD) are the average of the three measurements.

[0094] The protective effect of the sixth derivative on the RSL3-induced HT-22 hippocampal neuronal ferroptosis model: In some experiments, the neuroprotective activity of the derivatives was evaluated by comparing their ability to repair RSL3-induced HT-22 cell damage. The repair and protective effects of seven derivatives on damaged HT-22 cells were evaluated at concentrations of 15 μM, 50 μM, and 100 μM. Fer was used as a positive control in some experiments. As shown in Table 3-1, the derivatives exhibited varying degrees of repair activity against HT-22 cell damage at concentrations of 15 μM, 50 μM, and 100 μM, with the repair capacity increasing with increasing concentration. The sixth derivative, which was serotonin-amiditized, showed significant repair activity, reaching 93.52% at a concentration of 100 μM. The remaining derivatives showed relatively weak neuroprotective activity, with survival rates below 50% at concentrations of 15, 50, and 100 μM.

[0095] Based on the above experimental results, it was found that the sixth derivative has good activity and low toxicity in terms of neuroinflammation and neuroprotection. Therefore, the anti-inflammatory mechanism of the sixth derivative will be studied in depth.

[0096] Table 2. Neurorepair activity of NA and its P1-P7

[0097] d: Cell survival rate (n ≥ 4) after the compound repairs RSL3-induced HT-22 cell damage.

[0098] e: Positive control (Fer). All data (means ± SD) are the average of the five measurements.

[0099] Distribution of the sixth derivative in brain tissue: Reference Figure 2 To verify whether the sixth derivative can penetrate the mouse brain, a brain tissue distribution experiment was conducted. Mice were administered the sixth derivative (20 mg / kg) via gavage (po) and intraperitoneal injection (ip), with an equal volume of solvent (10% ethanol + 10% Tween 80 + 80% purified water) as a control (CON). Three hours after administration, hippocampal tissue was collected from mice for extraction and concentration. The drug content in the brain was determined using LC-MS / MS, and absolute quantification was performed based on a standard curve. The results showed that the sixth derivative can cross the blood-brain barrier (BBB) ​​in mice, therefore, the sixth derivative can be used for subsequent treatment of AD mice.

[0100] Effects of the sixth derivative on cognitive function in 5xFAD transgenic mice: In some experiments, 6-8 month old maternally related WT mice and 5xFAD mice were used and divided into a blank control group (CON, n=10), a blank drug treatment group (CON + sixth derivative, n=10), a model group (5xFAD, n=8), a nervonic acid group (5xFAD + NA-20, n=8), a low-dose nervonic acid sixth derivative group (5xFAD + sixth derivative-5, n=10), and a high-dose group (5xFAD + sixth derivative-20, n=9). After a 30-day drug intervention, the behavioral indicators of the mice were detected.

[0101] Novel Object Recognition (NOR) Experiment: like Figure 3 and Figure 4 As shown, there was no significant difference in the exploration time of the model group mice for new and old objects. After drug treatment, compared with the model group, the drug treatment group showed a significant enhancement in the ability to recognize new objects, indicating that the sixth derivative of compound can significantly improve the recognition and memory ability of mice.

[0102] Figure 3 and Figure 4 The resolution for new object recognition is denoted as . Statistical analysis was performed using two-way ANOVA and Newman-Keuls multiple comparison test; data are expressed as mean ± SEM (n=8~10). (***) p<0.001: highly significant difference; (##) p<0.01: compared with the blank control group; ns: no significant difference.

[0103] Y-maze experiment: like Figure 5 As shown, when the number of exploration arms was similar, the spontaneous alternation rate of mice in the drug-treated group was significantly higher than that in the model group, indicating that the sixth derivative can also significantly improve the short-term working memory of mice.

[0104] Figure 5 The spontaneous alternation rate of the Y-maze experiment is shown. Statistical analysis was performed using two-way ANOVA and Newman-Keuls multiple comparison test; data are expressed as mean ± SEM (n=8~10). (***) p<0.001: highly significant difference; (##) p<0.01: compared with the blank control group; ns: no significant difference.

[0105] Barnes Maze Experiment: like Figure 6 and Figure 7As shown, after training mice to remember the location of the target hole, a significant reduction in latency began to appear on the third and fourth days. Even after the target hole cassette was removed on the fifth day, the mice could still quickly find the location of the target hole. This indicates that the mice's spatial memory of the target hole was enhanced after drug treatment, and the spatial memory ability of the sixth derivative group was more significant than that of the raw material NA group.

[0106] Figure 6 and Figure 7 The escape latency period from day 1 to day 5 was shown. Statistical analysis was performed using two-way ANOVA and Newman-Keuls multiple comparison test; data are expressed as mean ± SEM (n=8–10). (*) p<0.05 indicates a significant difference, (**) p<0.01 indicates a highly significant difference, and (##) p<0.01 indicates a difference compared to the blank control group.

[0107] Experimental conclusions and discussion This study first explored a series of derivatizations based on nervonic acid, then systematically evaluated the nervonic acid derivatives, and explored the outstanding sixth derivative in depth.

[0108] In vitro experiments showed that the sixth derivative not only significantly inhibited NO release in an LPS-induced BV-2 microglia inflammation model (inhibition rate of 83.26% at 100 μM), but also exhibited outstanding repair capacity in an RSL3-induced HT-22 hippocampal neuronal ferroptosis model (cell survival rate of 93.52% at 100 μM), significantly outperforming other derivatives and the parent compound nervonic acid. Simultaneously, the sixth derivative showed no significant cytotoxicity at effective concentrations (survival rate >90%), indicating that it possesses both good safety and the potential for dual-target intervention in neuroinflammation and ferroptosis.

[0109] Tissue distribution experiments in mice demonstrated that the sixth derivative possesses excellent central nervous system-targeted delivery capabilities. Whether administered via gavage or intraperitoneal injection, the sixth derivative effectively penetrates the blood-brain barrier and enters the hippocampus, with intraperitoneal injection showing the best efficacy. This provides a pharmacokinetic basis for its central nervous system protective effects and subsequent administration methods.

[0110] In behavioral experiments involving new object recognition, the Y-maze, and the Barnes maze, the sixth derivative significantly improved cognitive dysfunction in 5xFAD transgenic mice. The working memory and spatial learning memory abilities of mice in the sixth derivative-treated group were significantly improved compared to the model group, and the high-dose group (20 mg / kg) showed better results than the parent compound nervonic acid. This indicates that the dual protective activity of the sixth derivative in vitro has successfully translated into an in vivo cognitive function repair effect.

[0111] This study focused on the structural modification and derivatization of nervonic acid. Through systematic evaluation and screening, the sixth derivative with the best activity was identified. Further investigation was conducted from three dimensions: in vitro activity, in vivo distribution, and pharmacodynamics, leading to the following conclusions: I. In vitro dual protective activity is significantly superior to that of the parent compound. The sixth derivative exhibited excellent dual activity against neuroinflammation and ferroptosis. In an LPS-induced BV-2 microglia inflammation model, this derivative significantly inhibited NO release, with an inhibition rate as high as 83.26% at a concentration of 100 μM. Simultaneously, in an RSL3-induced HT-22 hippocampal neuronal ferroptosis model, its repair capacity was outstanding, with a cell survival rate of 93.52% at a concentration of 100 μM. The improvement effects on these two key indicators were significantly better than other derivatives and the parent compound, nervonic acid. Furthermore, at the effective concentration, the cell survival rate remained above 90%, confirming its good cellular safety and potential for dual-target intervention against neuroinflammation and ferroptosis.

[0112] II. Possesses highly efficient central targeting delivery capabilities Tissue distribution experiments in mice confirmed that the sixth derivative possesses excellent blood-brain barrier penetration ability. Whether administered by gavage or intraperitoneal injection, this derivative effectively enters the hippocampus, with intraperitoneal injection showing the best brain penetration. This characteristic provides a solid pharmacokinetic basis for its neuroprotective effects in the central nervous system and optimizes the choice of administration route in subsequent experiments.

[0113] III. Significantly improves cognitive dysfunction in 5xFAD transgenic mice In in vivo pharmacodynamic evaluations, the sixth derivative successfully translated its in vitro activity into in vivo efficacy. In behavioral tests such as object recognition, the Y-maze, and the Barnes maze, this derivative significantly improved cognitive dysfunction in 5xFAD transgenic mice, and significantly enhanced working memory and spatial learning memory abilities in the model mice. Furthermore, under the same experimental conditions, the high-dose group (20 mg / kg) showed superior efficacy compared to the parent compound nervonic acid, fully demonstrating its potential application value as a novel neuroprotective agent.

[0114] In summary, the sixth derivative, as a novel nervonic acid derivative possessing both anti-neuroinflammatory and anti-ferroptosis activities, exhibits good blood-brain barrier permeability and effectively improves cognitive dysfunction in a 5xFAD mouse model. It is a potential anti-AD candidate compound with further development value. Future research can delve deeper into its molecular mechanisms regulating Aβ reduction and ferroptosis pathways, providing more substantial experimental evidence for its clinical application.

[0115] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this disclosure, and are not intended to limit them. Although this disclosure has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this disclosure.

Claims

1. A nervonic acid derivative, characterized in that, The nervonic acid derivative is a sixth derivative, and the chemical structure of the sixth derivative is as follows: 。 2. The nervonic acid derivative according to claim 1, characterized in that, The sixth derivative is a white solid with the following chemical structure: 。 3. The method for preparing the nervonic acid derivative according to claim 1, characterized in that, The nervonic acid derivative is a sixth derivative, and the preparation method includes the following steps: Nervonic acid, 1-hydroxybenzotriazole (HOBT) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were dissolved in dichloromethane and the carboxyl groups were activated by stirring at room temperature for 15 minutes. 4-(2-aminoethyl)benzene-1,2-diol hydrochloride and N,N-diisopropylethylamine (DIPEA) were added to the reaction system, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, the mixture was quenched with distilled water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain a white solid product.

4. The use of the nervonic acid derivative of claim 1 in the preparation of a medicament for the prevention or treatment of Alzheimer's disease.

5. The application according to claim 4, characterized in that, The drug exerts its therapeutic effect on Alzheimer's disease through a dual mechanism of inhibiting neuroinflammation and inhibiting hippocampal neuronal ferroptosis.

6. The application according to claim 4, characterized in that, The drug can penetrate the blood-brain barrier and accumulate in the hippocampus, improving cognitive impairment caused by Alzheimer's disease.

7. A pharmaceutical composition for treating Alzheimer's disease, characterized in that, The product comprises an effective dose of the nervonic acid derivative of claim 1, and a pharmaceutically acceptable carrier, excipient, or sustained-release agent.

8. The pharmaceutical composition according to claim 7, characterized in that, The dosage forms of the pharmaceutical composition include tablets, capsules, injections, microspheres, liposomes, or nanoparticles.

9. The use of the nervonic acid derivative according to claim 1 or 2 in the preparation of neuroinflammation inhibitors.

10. The use of the nervonic acid derivative according to claim 1 or 2 in the preparation of neuronal ferroptosis inhibitors.

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