Application of isocurcumenol in preparation of medicine for treating attention deficit hyperactivity disorder

Abstract

The invention relates to application of isocurcumenol in preparation of a medicine for treating attention deficit hyperactivity disorder, and belongs to the technical field of biological medicine. According to the invention, 1119 traditional Chinese medicine monomers are subjected to primary screening, secondary screening and dose dependence verification by using a per1b <- / -> zebra fish ADHD model. Through the multi-stage evaluation, it is determined that the isocurcumenol is a compound capable of remarkably rescuing ADHD related behavioral abnormalities. After treatment is carried out for 1 hour under the optimal concentration of 10 mu M, the isocurcumenol remarkably rescues hyperactivity, impulse and inattention existing in ADHD. The discovery shows that the isocurcumenol has a stable and repeatable behavior rescue effect, and supports the potential of the isocurcumenol as an anti-ADHD potential drug.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of isocurcumol in the preparation of drugs for the treatment of attention deficit hyperactivity disorder. Background Technology

[0002] Attention deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder. Clinically, ADHD has three main symptoms: hyperactivity, impulsivity, and inattention. These three conditions have detrimental effects on a child's intellectual, daily, and social development and may lead to secondary problems such as crime and drug abuse in adulthood. Individuals with ADHD exhibit hyperactivity, leading to sleep deprivation. ADHD has become a recognized disorder, and significant progress has been made in understanding its clinical manifestations, genetics, and treatment. ADHD is associated with functional and psychosocial comorbidities, including an increased risk of mood and anxiety disorders. Its prevalence is approximately 8% of the global population.

[0003] Uncontrolled ADHD leads to poor quality of life and an increased risk of suicide. While environmental factors are known to influence the development and manifestation of ADHD, genetic variation is considered a key etiology. Twin studies have played a crucial role in quantifying the heritability of ADHD, with meta-analyses of large-scale twin studies estimating the heritability of ADHD in childhood and adolescents to be between 0.7% and 0.8%. In contrast, the heritability in adults is estimated to be between 0.3% and 0.4%. However, some experts believe that the heritability in adults may be underestimated, and the true heritability is likely consistent with the estimated 0.7% to 0.8% in childhood. Identifying these heritable components deepens our understanding of the genetic basis of ADHD, particularly the role of genes related to neurotransmitters, including those regulating dopamine and serotonin.

[0004] Drug treatment for ADHD includes FDA-approved psychoactive agents that directly regulate neurotransmitters (such as methylphenidate and d-amphetamine derivatives), as well as non-psychoactive drugs that indirectly affect neurotransmitter activity and brain signal transduction (such as atomoxetine, clonidine, and guanosine), thereby influencing other pathways and downstream neurotransmitter responses. Cognitive behavioral therapy (CBT) is also used, especially in combination with medication. Although these methods can alleviate symptoms, current drug therapies have limited efficacy, produce significant adverse reactions, and often fail to resolve or prevent the disease. Summary of the Invention

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the problem that ADHD treatment targets in the prior art have significant adverse reactions.

[0006] To address the aforementioned technical problems, this invention provides the application of isocurcumol in the preparation of drugs for treating attention deficit hyperactivity disorder (ADHD), pointing to a new direction for the design of novel therapeutic agents and molecular targets for ADHD. This invention uses per1b - / - A total of 1119 traditional Chinese medicine monomers were initially screened, rescreened, and their dose-dependent effects were validated in a zebrafish ADHD model. Through this multi-stage evaluation, this invention identified isocurcumol as a compound that can significantly rescue ADHD-related behavioral abnormalities. After treatment at the optimal concentration of 10 μM for 1 hour, isocurcumol significantly rescued per1b. - / - Hyperactivity, impulsivity, and inattention. These findings suggest that isocurcumol has a stable and repeatable behavioral rescue effect, supporting its potential as a drug for treating ADHD.

[0007] The first objective of this invention is to provide the use of isocurcumol in the preparation of medicaments for the prevention or treatment of attention deficit hyperactivity disorder.

[0008] Furthermore, the structural formula of the isocurcumol is shown in Formula I:

[0009] .

[0010] Furthermore, the medication for the prevention or treatment of attention deficit hyperactivity disorder also includes pharmaceutically acceptable excipients.

[0011] Furthermore, the excipients include pharmaceutically acceptable salts, excipients, or carriers.

[0012] Furthermore, the excipients include at least one of the following: filler, binder, disintegrant, lubricant, flow aid, wetting agent, effervescent agent, colorant, sweetener, flavoring agent, preservative, dispersant, film-forming agent, plasticizer, pore-forming agent, opaque agent, retardant, and solvent.

[0013] Furthermore, the dosage form of the medication for the prevention or treatment of attention deficit hyperactivity disorder includes capsules, tablets, powders, injections, or oral preparations.

[0014] A second objective of this invention is to provide a treatment for attention deficit hyperactivity disorder (ADHD), the treatment comprising isocurcumol.

[0015] Furthermore, the effective concentration range of isocurcumol in the aforementioned attention deficit hyperactivity disorder (ADHD) treatment drug is 5-15 μM.

[0016] Furthermore, the aforementioned attention deficit hyperactivity disorder (ADHD) treatment drug upregulates the expression of adrenergic receptor β1 (adrb1), neuronal PAS domain protein 4a (npas4a), serotonin receptor 1b (htr1b), adrenergic receptor β2 (adrb2b), and dopamine receptor D2a (drd2a).

[0017] A third objective of this invention is to provide the application of isocurcumol in the preparation of a neuroactive ligand-receptor interaction pathway activator, wherein the neuroactive ligand-receptor interaction pathway activator upregulates the expression of adrb1, npas4a, htr1b, adrb2b, and drd2a.

[0018] The fourth objective of this invention is to provide the application of isocurcumol in the preparation of a steroid hormone metabolic pathway activator, which upregulates the expression of steroid 5α-reductase 1 (srd5a1), UDP glucuronyl transferase family 1 member A1 (ugt5a1), and hydroxysteroid 11-β dehydrogenase 2 (hsd11b2).

[0019] Compared with the prior art, the above-described technical solution of the present invention has the following advantages:

[0020] This invention discovers that isocurcumol is a compound that can significantly rescue ADHD-related behavioral abnormalities. After treatment with the optimal concentration of 10 μM for 1 hour, isocurcumol significantly rescued per1b. - / - Hyperactivity, impulsivity, and inattention. These findings suggest that isocurcumol has a stable and repeatable behavioral rescue effect, supporting its potential as a drug for treating ADHD. Attached Figure Description

[0021] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0022] Figure 1 The results show the test results of ADHD-like behaviors in each group. A is a graph showing the relationship between the dose-dependent effect of isocurcumol and the total movement distance (mm), and B is a graph showing the comparison between isocurcumol and the positive control. **** indicates P<0.0001, *** indicates P<0.001, ** indicates P<0.01, * indicates P<0.05, and ns indicates no significance.

[0023] Figure 2 This is a time-process graph showing the average distance traveled during the impulsivity measurement for each group;

[0024] Figure 3 This is a diagram showing the results of an experiment involving zebrafish color preference and a maze.

[0025] Figure 4 This is a graph showing the molecular validation results of isocurcumenol in an ADHD zebrafish model. A is the KEGG pathway enrichment analysis, B is the GO enrichment result graph, C is the expression result of key neurotransmitter receptor genes in different treatment groups, and D is the relative expression result of adra1ab, drd4 and gabra6a genes in different treatment groups. Isocurcumenol is abbreviated as iso.

[0026] Figure 5 This is a graph showing the expression analysis of steroid hormone-related genes. Detailed Implementation

[0027] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0028] Example

[0029] 1. Animals

[0030] Use wild-type and per1b 6-12 month old - / - Adult mutant zebrafish. Maintain a temperature of 28.5±1°C and a pH of 7.2-7.6 under a 14-hour light / 10-hour dark cycle (14L:10D). Feed brine shrimp twice daily (morning and evening), and dry food at midday.

[0031] 2. Collection of zebrafish embryos and juveniles

[0032] Adult zebrafish were placed in a mating box, with males and females separated by a baffle. The next morning, the baffle was removed after the lights were turned on, and embryos were collected one hour later. Unfertilized embryos were removed, and the remaining embryos were cultured in E3 medium at 28.5°C under 14L:10D illumination until they reached 6 days post-fertilization.

[0033] 3. Behavior of juvenile fish

[0034] Behavioral analysis was performed using a zebrafish behavior analyzer (ViewPoint) equipped with an infrared high-speed camera. Six-day-old post-fertilization per1b mutant larvae were placed in 96-well plates, each containing 200 μL of E3 medium. The larvae were pretreated with the test compound at a final concentration of 10 μM for 1 hour before being transferred to the behavior monitoring system. Zebrafish movement was recorded continuously for 1 hour under constant light (300 lux).

[0035] 4. Drug screening

[0036] The screening used TargetMol's herbal monomer library (L6810). Stock solutions were prepared in DMSO at 10 mM. The procedure is as follows:

[0037] (1) Add one juvenile fish to each well of a 96-well plate containing 200 μL of E3 medium.

[0038] (2) 0.2 μL of the drug stock solution was administered to 200 μL to achieve a final concentration of 10 μM. At the same time, an appropriate control group was set up, including a negative control with only DMSO added and an untreated per1b mutant control.

[0039] (3) One hour after exposure to the drug, the juvenile fish were transferred to a behavior monitoring system and their movement was recorded for one hour.

[0040] 5. Results and Statistical Analysis

[0041] The experimental data were statistically analyzed using GraphPad Prism8. The bar chart data were labeled as mean ± SEM. The experimental results were analyzed using a one-way value <0.05, which means that there are significant differences between the data, indicated by *; when P <0.01, it means that there are significant differences between the data, indicated by **; when P <0.001, it means that there are very significant differences between the data, indicated by ***.

[0042] 6. Screening of monomeric compounds of traditional Chinese medicine in per1b knockout ADHD models

[0043] In this embodiment, a library of 1119 monomeric compounds from traditional Chinese medicine was used to treat per1b at an initial concentration of 20 μM. - / - Zebrafish fry. Behavioral assessment was performed 1 hour after drug treatment, with a focus on hyperactivity in fry 6 days post-fertilization. Screening showed that isocurcumenol had a significant rescue effect on hyperactivity-like behaviors.

[0044] 7. The discovery of isocurcumol as an effective anti-ADHD compound

[0045] To confirm the rescue effect, an impulsive behavior test was performed on isocurcumol. The results showed that at a concentration of 10 μM, isocurcumol exhibited the most significant rescue effect, almost completely rescuing per 1b. - / - The ADHD-like behaviors of the juvenile fish returned to normal levels.

[0046] To further verify the role of isocurcumol in attention deficit hyperactivity disorder (ADHD), an experimental group design was carried out according to Table 1.

[0047] Table 1 Grouping and Related Processing

[0048]

[0049] Each group of zebrafish juveniles was treated for 1 hour, then exposed to alternating light (90 seconds) and dark (90 seconds) phases for a total experimental time of 1 hour to assess the activity levels of different groups of zebrafish throughout the entire period of impulsive behavior. Results are as follows: Figure 1 As shown in Figure A, per1b - / - The hyperactive phenotype in juvenile fish was almost completely recovered in a dose-dependent manner, with 10 μM isocurcumol showing the best rescue effect. Furthermore, the rescue effect of 10 μM isocurcumol was similar to that of atoxetine, a drug already in clinical use. Figure 1 B). Furthermore, per1b - / - Impulsivity in juvenile fish decreased significantly in a dose-dependent manner. Figure 2 ).

[0050] In addition, this embodiment uses a custom-designed multi-color discrimination device to assess the attention deficit of zebrafish juveniles. The device consists of six independent cross-shaped modules, each providing four different color zones (red, blue, black, and yellow). The device structure is as follows: Figure 3 (As shown in A). Previous studies have shown that zebrafish avoid yellow, making it a stable negative preference control. In per1b - / - In a zebrafish ADHD model, juvenile fish exhibited abnormally frequent entry into the yellow region, indicating impaired attention-based decision-making. Behavioral tracking was performed for one hour after exposure to isocurcumol or DMSO to quantify color region selection, switching frequency, and decision delay. Results are as follows: Figure 3 As shown in B, treatment with 10 μM isocurcumol significantly reduced per1b. - / - The juvenile fish reverted to the avoidance pattern observed in the yellow area, reverting to the pattern seen in wild-type fish. Figure 3 B). These results demonstrate that isocurcumol can significantly reduce core behavioral abnormalities associated with ADHD in a zebrafish model, including all core symptoms (hyperactivity, impulsivity, and inattention), highlighting its potential as a neuroactive therapeutic agent.

[0051] 8. Changes in transcriptional levels in zebrafish treated with isocurcumol

[0052] RNA sequencing analysis showed that isocurcumol significantly regulated multiple signaling pathways, particularly the neuroactive ligand-receptor interaction pathway and other neurotransmitter-related pathways. Figure 4 A). GO enrichment analysis showed that differentially expressed genes are mainly involved in synaptic signal transduction, neurotransmitter receptor activity, and neural signal transduction processes. Figure 4B). Further analysis revealed abnormal expression of multiple ADHD-related neurotransmitter genes in the disease model, which showed an overall improvement trend after treatment with isocurcumol, including genes related to dopaminergic, serotonergic, and adrenergic signaling. Figure 4 C). Quantitative PCR was used to verify the selected genes, and the results were consistent with RNA sequencing data, confirming that isocurcumol can regulate key genes in the neurotransmitter signaling pathway. Figure 4 D).

[0053] RNA-seq data showed that in a zebrafish model of ADHD, isocurcumin treatment regulated genes related to steroid hormone metabolism. Specifically, isocurcumin treatment restored the expression of steroid 5α-reductase 1 (srd5a1) and its downstream steroid metabolism-related genes, such as polypeptide B1 (ugt1b2) of the UDP-glucuronyltransferase 2 family, UDP-glucuronyltransferase family 1 member A1 (ugt5a1), and hydroxysteroid 11-β dehydrogenase 2 (hsd11b2). Figure 5 These genes are all part of zebrafish orthologous gene dre:100384893 and steroid hormone biosynthesis pathway (KO00140).

[0054] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. Application of isocurcumol in the preparation of drugs for the prevention or treatment of attention deficit hyperactivity disorder.

2. The application according to claim 1, characterized in that, The medications for the prevention or treatment of attention deficit hyperactivity disorder also include pharmaceutically acceptable excipients.

3. The application according to claim 2, characterized in that, The excipients include pharmaceutically acceptable salts, excipients, or carriers.

4. The application according to claim 3, characterized in that, The excipients include at least one of the following: filler, binder, disintegrant, lubricant, flow aid, wetting agent, effervescent agent, colorant, sweetener, flavoring agent, preservative, dispersant, film-forming agent, plasticizer, pore-forming agent, opaque agent, retardant, and solvent.

5. The application according to claim 1, characterized in that, The dosage forms of the medications for the prevention or treatment of attention deficit hyperactivity disorder include capsules, tablets, powders, injections, or oral dosage forms.

6. A medication for treating attention deficit hyperactivity disorder, characterized in that, The medications used to treat attention deficit hyperactivity disorder include isocurcumol.

7. The medication for treating attention deficit hyperactivity disorder according to claim 6, characterized in that, The effective concentration range of isocurcumol in the aforementioned medication for treating attention deficit hyperactivity disorder is 5-15 μM.

8. The medication for treating attention deficit hyperactivity disorder according to claim 6, characterized in that, The medication for treating attention deficit hyperactivity disorder upregulates the expression of adrenergic receptor 1, neuronal PAS domain protein 4, 5-hydroxytryptamine 1B receptor, adrenergic receptor 2, and dopamine receptor D2.

9. The application of isocurcumol in the preparation of activators of neuroactive ligand-receptor interaction pathways, characterized in that, The neuroactive ligand-receptor interaction pathway activator upregulates the expression of adrenergic receptor 1, neuronal PAS domain protein 4, 5-hydroxytryptamine 1B receptor, adrenergic receptor 2, and dopamine receptor D2.

10. The application of isocurcumol in the preparation of steroid hormone metabolic pathway activators, characterized in that, The steroid hormone metabolic pathway activator upregulates the expression of steroid 5α-reductase 1, UDP glucuronyltransferase 2 family member B1, UDP glucuronyltransferase 1 family member A1, and hydroxysteroid 11-β dehydrogenase 2.

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