Application of longishin in preparation of medicine for preventing or treating jetlag syndrome

Abstract

The invention relates to an application of longishin (CRS, C17H16N2O2S) in preparation of a medicine for preventing or treating a time difference syndrome, and belongs to the technical field of biological medicines. The invention finds that the longiday element can adjust the biological clock phase of the zebra fish at 0.3-80 [mu] M. Furthermore, the action mechanism of the longdate element in the period and phase of the biological clock is defined, the longdate element is found to adjust the phase of the biological clock instead of mainly aiming at sleep, the compound has more and more obvious phase adjustment along with the increase of concentration, and the compound is especially suitable for people who intercontinental cross-time-zone travel, and has a wide application prospect. And the internal biological clock can be properly adjusted by adopting corresponding concentration according to the change amount of the time zone, so that the clock can quickly adapt to the local time. Meanwhile, the biological clock regulation function of the longbiantin is reversible, the longbiantin has the regulation function after being taken, and the longbiantin recovers to a normal biological clock after being canceled.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of long-day extract in the preparation of drugs for the prevention or treatment of jet lag syndrome. Background Technology

[0002] The Earth's rotation creates a 24-hour biological clock. Shift work, intercontinental travel across time zones, irregular eating and sleeping habits, and genetic mutations related to the biological clock can all lead to varying degrees of circadian rhythm disruption. Circadian rhythm disruption affects the development of tumors, cardiovascular diseases, metabolic diseases, mental illnesses, reproductive health, and thus impacts human health. In my country, approximately 10-15 million individuals experience significant jet lag each year, and this number rises to 20-30 million per year if estimated based on travel frequency. This group is primarily concentrated among residents of first- and second-tier cities who frequently travel internationally, business people, students studying abroad, aviation professionals, diplomats, and employees of multinational corporations. In the context of globalization, cross-border remote collaboration is increasing; even without travel, working with colleagues in different time zones at night can lead to "social jet lag." After flying across time zones, approximately 70%-80% of people experience varying degrees of jet lag symptoms (fatigue, insomnia, decreased concentration, gastrointestinal discomfort, headaches, etc.), with about 30% experiencing significant symptoms, termed jet lag syndrome.

[0003] Currently, there is no specific cure for jet lag. Symptoms can only be alleviated with sleep-regulating medications or supplements such as melatonin, including melatonin supplements like tasematron and ramelton, sleep aids like diphenhydramine and zolpidem, and wakefulness-promoting drugs like caffeine and modafinil. All of these medications regulate sleep / wakefulness. However, the discomfort caused by jet lag is mainly due to the discrepancy between the body's own biological clock and the light and dark times of the external environment, resulting in a lack of coordination between the internal biological clock and external time. Adjusting sleep alone cannot completely synchronize the body's internal biological clock with environmental time in a short period. Furthermore, sleep-promoting / wakefulness drugs also have certain side effects. For example, diphenhydramine's side effects include drowsiness the next day, slowed reaction time, and impaired coordination; zolpidem may cause taste abnormalities; tasematron and ramelton may cause headaches and dizziness; caffeine may cause palpitations and increased blood pressure; and modafinil may cause anxiety, irritability, headaches, and dizziness. Therefore, there is an urgent need to develop new anti-jet lag drugs with fewer toxic side effects that are specifically designed to adjust the biological clock. Summary of the Invention

[0004] Therefore, the technical problem to be solved by the present invention is to overcome the lack of an effective drug for improving jet lag in the prior art.

[0005] To address the aforementioned technical problems, this invention provides the application of long-day glycosides (CRS) in the preparation of drugs for the prevention or treatment of jet lag. This invention utilizes the diurnal behavior of zebrafish, which is similar to that of humans, to screen for drugs that can affect the cycle and phase of the biological clock, ultimately discovering long-day glycosides (CRS, molecular formula: C). 17 H 16 N2O2S can adjust the circadian rhythm phase in zebrafish at concentrations of 0.08-80 μM. Furthermore, this invention clarifies the mechanism of action of long-day agonist in the circadian rhythm cycle and phase, revealing that long-day agonist regulates the circadian rhythm phase, rather than primarily targeting sleep. Moreover, the phase-regulating effect of this compound becomes increasingly significant with increasing concentration, making it particularly suitable for individuals traveling across continental time zones. The appropriate concentration can be used to adjust their internal circadian rhythm according to the degree of time zone change, allowing them to quickly adapt to the local time. Simultaneously, the circadian rhythm regulation function of long-day agonist is reversible; the regulatory function is present after taking the compound, and the circadian rhythm returns to normal after discontinuation.

[0006] The first objective of this invention is to provide the use of long-day extract in the preparation of medicaments for the prevention or treatment of jet lag syndrome.

[0007] Furthermore, the structural formula of the long-day element is shown in Formula I:

[0008] .

[0009] Furthermore, the jet lag prevention or treatment medication also includes pharmaceutically acceptable excipients.

[0010] Furthermore, the excipients include pharmaceutically acceptable salts, excipients, or carriers.

[0011] 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.

[0012] Furthermore, the dosage form of the jet lag prevention or treatment drug includes capsules, tablets, powders, injections, or oral dosage forms.

[0013] A second objective of this invention is to provide a product for the prevention or treatment of jet lag syndrome, the product comprising a long-lasting nutrient.

[0014] Furthermore, the effective concentration range of the long-term effect factor in the jet lag prevention or treatment product is 0.08-80 μM.

[0015] A third objective of this invention is to provide the application of long-day sedative in the preparation of a medicament for regulating the phase and / or amplitude of the biological clock.

[0016] A fourth objective of this invention is to provide a drug for regulating the phase and / or amplitude of the biological clock, said drug comprising long-day hormone.

[0017] Furthermore, the drug can be used to treat jet lag, shift work, age-related sleep disorders, or circadian rhythm-related sleep disorders.

[0018] Compared with the prior art, the above-described technical solution of the present invention has the following advantages:

[0019] This invention discovers long-day element (molecular formula: C 17 H 16 N2O2S can adjust the circadian rhythm phase in zebrafish at concentrations of 0.08-80 μM. Furthermore, this invention reveals that long-term circadian rhythm regulation primarily targets the phase of the circadian rhythm, rather than sleep, and that the phase-regulating effect becomes increasingly significant with increasing concentration. Therefore, long-term circadian rhythm regulation is suitable for individuals traveling across continental time zones, allowing them to adjust their internal circadian rhythm to adapt quickly to local time by using appropriate concentrations based on the degree of time zone change. Simultaneously, the circadian rhythm regulation function of long-term circadian rhythm regulation is reversible; the regulatory function is present after taking the compound, and the circadian rhythm returns to normal after discontinuation. Attached Figure Description

[0020] 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.

[0021] Figure 1 This is the drug screening process that affects the behavioral rhythms of zebrafish;

[0022] Figure 2 The behavioral characteristics of zebrafish treated with long-day saturation inhibitor (CRS) under DD conditions, (AB) the behavioral rhythms of wild-type juvenile zebrafish treated with several low (A) and high (B) concentrations of CRS under DD conditions;

[0023] Figure 3 This is a bioluminescence assay of the Tg(per3:luc) characteristics of zebrafish treated with CRS.

[0024] Figure 4After washing away CRS, the phase delay effect disappeared. (A) Zebrafish were treated with three concentrations of long-day diathermycin for two days. After washing away the long-day diathermycin, the bioluminescence rhythm was monitored for five days under DD conditions. (BD) Bioluminescence rhythms under DD conditions with long-day diathermycin concentrations of 0.3 μM (B), 1.25 μM (C), and 5 μM (D) during and after treatment.

[0025] Figure 5 Other casein kinase inhibitors failed to regulate the behavioral rhythms of zebrafish under DD conditions. (AD) Under DD conditions, the behavioral rhythms of wild-type juvenile zebrafish were treated with 10 μmol of WAY-650313 (A), WAY-621640 (B), GO289 (C) and Casein kinase 1δ-IN-25 (D).

[0026] Figure 6 This is a structural similarity comparison between long-terminus and WAY-650313, WAY-621640, GO289, and Casein Kinase 1δ-IN-25. 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 1

[0029] 1. Animals

[0030] Wild-type zebrafish aged 6-12 months. They are raised using a professional breeding system, with the water temperature maintained at 28.5±1℃ and the pH between 7.2 and 7.6. The light cycle in the fish room is 14 hours of light / 10 hours of darkness, i.e., 14L:10D. They are fed brine shrimp in the morning and evening and dry food at noon.

[0031] 2. Obtaining zebrafish embryos and juveniles

[0032] On the first night, a pair of adult zebrafish, male and female, were placed in a mating box, separated by a partition. The next day, after the aquarium lights were turned on, the partition was removed, and the embryos were collected one hour later and placed in E3 culture medium. Unfertilized embryos were removed, and the remaining embryos were placed in a light-dark incubator (14L:10D, 300 lux) at 28.5℃. The zebrafish fry were ready for use when they reached 6-7 days post-fertilization (dpf).

[0033] 3. Behavior of juvenile fish

[0034] The experimental apparatus used was a zebrafish behavior analyzer (Noldus). The instrument contained a high-speed infrared camera to capture the movement parameters of the juvenile fish in each well. At 5 dpf, zebrafish juveniles were added to 96-well square plates, with only one fish per well. After abstaining from water, 650 μL of E3 juvenile culture medium containing 10 μM of the target compound was added. The 96-well plate was then transferred to a zebrafish behavior tracking system, and continuous monitoring was performed for 72 hours under dark conditions, with data recording intervals set at 10-minute intervals.

[0035] 4. Drug screening

[0036] The experiment used a known active library (L2000) from Selleck. All drugs used were dissolved in 100 μL of dimethyl sulfoxide (DMSO) at a concentration of 10 mM. In one experiment, each compound was used to treat eight juvenile zebrafish; that is, one 96-well plate could screen the effects of 11 compounds (excluding the DMSO control group) on zebrafish behavioral rhythms. To ensure the accuracy of the screening results, compounds that showed significant effects on behavioral rhythms in the first round of screening were subjected to a second and third round of screening for confirmation. Finally, compounds with consistent data from all three rounds of screening were selected for further experimental verification.

[0037] 5. Long-term in vivo monitoring of bioluminescence rhythms

[0038] Tg(per3:luc) zebrafish larvae were kept under a 14-hour light / 10-hour dark cycle at 28.5°C, with the light cycle starting at 9:00 AM and ending at 11:00 PM. On day 5 post-fertilization, individual Tg(per3:luc) larvae were placed in 96-well plates containing 200 μL of system water and 0.5 mmol / L D-luciferin potassium salt. 10 mmol / L stock solution (dissolved in dimethyl sulfoxide) was added to each well by pipetting to introduce the small molecule compound. Before the experiment, the culture plates were sealed with a transparent sealing film to reduce liquid evaporation. Bioluminescence was monitored using a Luminoskan Ascent microplate bioluminescence analyzer for at least 72 hours at 6-12 minute intervals.

[0039] 6. Results and Statistical Analysis

[0040] Rhythm analysis was performed using the Biodare2 platform (https: / / biodare2.ed.ac.uk / ). The JTK (Jonckheere-Terpstra-Kendall) periodicity algorithm was employed, using a pre-defined COS_2H waveform. For periodicity analysis, the input data was not detrended, with an expected period range of 18 to 34 hours, and the MFourFit method was used. The term Δperiod refers to the difference in period between the compound treatment group and the corresponding batch of DMSO treatment group. Similarly, Δphase refers to the phase difference between the compound treatment group and the corresponding batch of DMSO treatment group within a 24-hour time frame. Δamplitude represents the ratio of the amplitude of the compound treatment group to the amplitude of the corresponding batch of DMSO treatment group. When expressed in ns, it indicates no significant difference between the data; when P < 0.05, it indicates a significant difference between the data, indicated by *; when P < 0.01, it indicates a relatively significant difference between the data, indicated by **; when P < 0.001, it indicates a highly significant difference between the data, indicated by ***.

[0041] Example 2

[0042] 1. Drug screening using wild-type zebrafish juveniles with compounds from a known active library.

[0043] We treated 5-day-old post-fertilization zebrafish larvae with 10 μM of a known active compound around 5 PM on the screening day, and then quickly placed them in a behavior analyzer for continuous 72-hour behavioral monitoring under dark conditions. A DMSO negative control group was included in each 96-well plate. The behavioral curves obtained for each compound treatment were compared with those for the DMSO group, and the period, phase, and amplitude were calculated using software. Figure 1 The screening process consisted of three rounds. The first two rounds were conducted through behavioral screening, and the third round was validated using the Tg(per3:luc) fish strain. The long diurnal compound was identified as a compound that significantly affected the phase in all three rounds of screening.

[0044] 2. Determine the effective concentration of long-day factor that affects the phase of the biological clock.

[0045] To further clarify the effective concentration of long-day diathermy influencing the biological clock, we treated zebrafish juveniles with 14 concentrations of long-day diathermy ... Figure 2 The results showed that phase lag began at a concentration of 0.08 μM, and the degree of phase lag increased with increasing concentration, reaching a plateau at 20 μM. Importantly, the rhythmic pattern was still present. Figure 2 Similar phenotypes were also observed in zebrafish exposed to seven concentrations of Tg(per3:luc) ranging from 1.25 to 80 μM. Figure 3After two days of treatment with long-day compound (LTC) on Tg(per3:luc), the compound was removed at 7 dpf, leading to a significant recovery of the circadian rhythm under subsequent five days of DD conditions. Figure 4 This indicates that the effects of long-day hormone are reversible, and the maintenance of the altered phenotype depends on the continued presence of the compound. Therefore, long-day hormone has a wide effective concentration range for regulating the circadian rhythm, from 0.08 μM to 80 μM, and the higher the concentration, the more pronounced the delay in circadian rhythm regulation. Furthermore, the regulation of the circadian rhythm by long-day hormone is controllable, and the circadian rhythm returns to normal after drug withdrawal.

[0046] 3. Long-terminant regulates the phase of the biological clock by targeting CK1δ.

[0047] Structural comparisons revealed that long-day kinase inhibitor (CK1) is an inhibitor of casein kinase 1 (CK1). We compared the effects of other casein kinase inhibitors on the behavioral rhythms of zebrafish under continuous darkness conditions. Figure 5 ), including WAY-650313 (CK1δ inhibitor, Figure 5 A) WAY-621640 (CK1δ inhibitor, Figure 5 B) GO289 (CK2 inhibitor) Figure 5 C) Caseinkinase 1δ-IN-25 (CK1δ inhibitor) Figure 5 (D) None of the four compounds at 10 μM affected behavioral rhythms. Although the targets were the same or similar, the effects varied greatly, further demonstrating the superiority of long-day cyclin in regulating jet lag. To analyze the reasons for the significant differences in the effects of different CK1 inhibitors on circadian rhythm regulation, we compared... Figure 5 The structural formulas of four compounds and chronoside were compared. The results showed that the structural similarity between the four compounds themselves and with chronoside was not high. CK1 can phosphorylate phosphorylation sites of multiple genes, and in addition to core circadian rhythm genes, it may also regulate multiple pathways. Figure 6 Therefore, it is speculated that the above four casein kinase inhibitors did not affect the phosphorylation of core circadian rhythm proteins. However, long-day kinase inhibitors can affect core circadian rhythm proteins, thereby playing a role in regulating circadian rhythm phase.

[0048] 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 long-day extract in the preparation of drugs for the prevention or treatment of jet lag syndrome.

2. The application according to claim 1, characterized in that, The medications for the prevention or treatment of jet lag 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 jet lag include capsules, tablets, powders, injections, or oral medications.

6. A product for the prevention or treatment of jet lag syndrome, characterized in that, The jet lag prevention or treatment products include Changrisu.

7. The jet lag prevention or treatment product according to claim 6, characterized in that, The effective concentration range of the long-term effect factor in the aforementioned time lag prevention or treatment product is 0.08-80 μM.

8. Application of long-day sedative in the preparation of drugs that regulate the phase and / or amplitude of the biological clock.

9. A drug for regulating the phase and / or amplitude of the biological clock, characterized in that, The drug includes chlorhexidine.

10. The medicament according to claim 9, characterized in that, The drug can be used to treat jet lag, shift work, age-related sleep disorders, or circadian rhythm-related sleep disorders.

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