Application of edaravone in prevention and treatment of cognitive impairment caused by plateau hypoxia environment

Abstract

The invention relates to an application of edaravone in prevention and treatment of cognitive impairment caused by plateau hypoxia environment, in particular to an application of edaravone or pharmaceutically acceptable salts or analogues or derivatives thereof in preparation of drugs for prevention and / or treatment of cognitive impairment caused by plateau hypobaric hypoxia. The invention surprisingly discovers that edaravone has a very good prevention and treatment effect on cognitive impairment caused by plateau hypobaric hypoxia, and particularly has an outstanding treatment effect on cognitive impairment caused by chronic plateau hypobaric hypoxia; biochemical indexes, hippocampal neurological defects, neuroinflammation and the like caused by chronic plateau hypobaric hypoxia can be improved.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceuticals, and particularly relates to a new use of edaravone, and more particularly to the application of edaravone in the preparation of a drug for preventing and / or treating cognitive dysfunction caused by high altitude hypobaric hypoxia. Background Art

[0002] High altitude hypobaric hypoxia (HAH) refers to a situation in high altitude areas where the partial pressure of oxygen decreases due to the lower atmospheric pressure, resulting in a lack of sufficient oxygen in the body. The higher the altitude, the lower the atmospheric pressure, and the decrease in air pressure leads to a reduction in the partial pressure of oxygen, causing the respiratory system to work harder to supply sufficient oxygen to various parts of the body, thereby causing a series of physiological and symptomatic changes. The brain is an organ with high metabolism and oxygen consumption and is extremely vulnerable to the effects of hypoxia. Chronic hypobaric hypoxia can damage the cognitive functions of humans and animals, especially learning and memory abilities. The incidence of cognitive impairment in the elderly living in high altitude areas is approximately twice that in other regions of the world. In addition, even when returning to plain areas, the cognitive dysfunction caused by hypobaric hypoxia persists.

[0003] To alleviate or treat cognitive dysfunction caused by high altitude hypoxia, scholars at home and abroad have conducted a large number of studies. Disappointingly, there is currently no effective drug to treat or prevent cognitive impairment caused by high altitude hypobaric hypoxia, especially in long-term hospitalized patients. Oxygen therapy is an effective treatment method, but most people cannot use it. Many drugs, such as the corticosterone synthesis inhibitor metyrapone and isradipine, are not suitable for long-term use due to potential side effects.

[0004] There is a need in this field for a drug that can prevent and / or treat cognitive dysfunction caused by high altitude hypobaric hypoxia. Summary of the Invention

[0005] The purpose of the present invention is to provide a new use of edaravone, that is, for preventing and / or treating cognitive impairment caused by high altitude hypobaric hypoxia.

[0006] First of all, the present invention provides the application of edaravone or its pharmaceutically acceptable salt or analogue or derivative (preferably alone as the sole active ingredient) in the preparation of a drug for preventing and / or treating cognitive impairment caused by high altitude hypobaric hypoxia.

[0007] On the other hand, the present invention also provides a pharmaceutical composition for preventing and / or treating cognitive impairment caused by high altitude hypobaric hypoxia, comprising edaravone or its pharmaceutically acceptable salt or analogue or derivative, and one or more pharmaceutically acceptable excipients.

[0008] According to the present invention, the administration route of the pharmaceutical composition is intravenous drip, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or intravaginal route. Preferably, the administration route of the pharmaceutical composition is oral administration.

[0009] In one embodiment of the present invention, the pharmaceutical dosage form for oral administration is a solid preparation, preferably a solid dispersion preparation, which comprises the active ingredient edaravone or a pharmaceutically acceptable salt thereof, the polymer carrier is selected from Soluplus, polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose acetate (HPMCAS), hydroxypropyl cellulose (HPC), and chitosan (wherein Soluplus and / or HPMC are preferred, and Soluplus is more preferred), and optionally a surfactant such as TPGS 1000. In the pharmaceutical composition, the unit dose of edaravone is 0.001 - 1000 mg, preferably 0.01 - 500 mg, more preferably 0.1 - 100 mg, most preferably 1 - 50 mg, such as 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, and 50 mg.

[0010] The present invention relates to a method for preventing and / or treating cognitive impairment caused by hypobaric hypoxia at high altitude using edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof, which comprises administering a therapeutically effective amount of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof to a subject in need.

[0011] The present invention also relates to edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof, as a medicament for preventing and / or treating cognitive impairment caused by hypobaric hypoxia at high altitude, which medicament comprises edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof.

[0012] The present invention unexpectedly discovers that edaravone has very good preventive and therapeutic effects on cognitive impairment caused by hypobaric hypoxia at high altitude, especially has outstanding therapeutic effects on cognitive dysfunction caused by chronic hypobaric hypoxia at high altitude, and can improve biochemical indexes, hippocampal nerve defects, neuroinflammation, etc. caused by chronic hypobaric hypoxia at high altitude. Brief Description of the Drawings

[0013] In order to more clearly describe the technical solutions of the present invention, a brief introduction will be given below in conjunction with the drawings. Obviously, these drawings are only some specific embodiments recorded in the present application. The present invention includes but is not limited to these drawings.

[0014] Figure 1 Showing a schematic diagram of the Barnes maze;

[0015] Figure 2Shows the representative trajectories of mice in the open field test;

[0016] Figure 3 Shows the total movement distance of mice in the open field test;

[0017] Figure 4 Shows the representative heatmap of mice in the novel object recognition test (N: novel object; F: familiar object);

[0018] Figure 5 Shows the discrimination rate of mice in the novel object recognition test;

[0019] Figure 6 Shows the latency of mice to enter the target hole in the Barnes maze test;

[0020] Figure 7 Shows the errors made by mice during learning in the Barnes maze test;

[0021] Figure 8 Shows the SOD and GSH / GSSG levels of mice;

[0022] Figure 9 Shows the number of Iba1+ cells in mice;

[0023] Figure 10 Shows the number of neural progenitor cells (Sox2+ / GFAP+) in the DG region of the hippocampus of mice;

[0024] Figure 11 Shows the number of DCX+ cells in the DG region of the hippocampus of mice. Detailed implementation mode

[0025] The present application will be further described in detail below with reference to the accompanying drawings and embodiments. Through these descriptions, the features and advantages of the present application will become more clearly defined.

[0026] The term "pharmaceutically acceptable" means approved by a regulatory agency such as the EMEA (Europe) and / or the FDA (US) and / or any other national regulatory agency for use in animals and preferably in humans.

[0027] In addition, the technical features involved in different embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0028] Definition

[0029] The term "pharmaceutically acceptable" means approved by a regulatory agency such as the EMEA (Europe) and / or the FDA (US) and / or any other national regulatory agency for use in animals and preferably in humans.

[0030] The term "excipient" refers to a diluent, adjuvant or carrier administered together with a therapeutic agent. Examples of suitable pharmaceutical excipients are described in "Remington’s Pharmaceutical Sciences" written by E.W. Martin.

[0031] 1. Edaravone and its salts, analogs or derivatives

[0032] Edaravone (EDA) is one of the pyrazolone derivatives, and its chemical name is 3-methyl-1-phenyl-2-pyrazolin-5-one, with the molecular formula C 10 H 10 N2O, and the structural formula is shown as follows:

[0033]

[0034] The core structure or active center of EDA is 2-pyrazolin-5-one, which plays a major role in free radical scavenging activity.

[0035] EDA is a nitrogen-containing heterocyclic compound, which is basic and easily reacts with acids to form salts. Its pharmaceutically acceptable salts include common inorganic acid salts and organic acid salts in the art. Among them, inorganic acid salts include hydrochloride, carbonate, phosphate, etc., and organic acid salts include citrate, tartrate, acetate, oxalate, salicylate, malate, lactate, etc.

[0036] EDA analogs include that the 3-position methyl on the pyrazoline ring can be a lower (C 1-6 ) alkyl such as ethyl, propyl, or a lower alkoxy group such as methoxy, ethoxy, etc.; or the 3-position methyl can be H, and the H at the 4-position can be replaced by a lower alkyl or alkoxy group. Derivatives of edaravone include esters, that is, the 5-position ketone of the pyrazoline ring is converted into enol and reacts with carboxylic acid to form esters, such as methyl ester, ethyl ester, etc. After hydrolysis in vivo, the ester (precursor) is then converted back into a ketone. In addition, the phenyl group is also optionally substituted by one or more substituents selected from lower alkyl, lower alkoxy, nitro, halogen, etc.

[0037] EDA derivatives include up to 18 EDA derivatives disclosed in Table 1-2 of Bioorg. Med. Chem. Lett. 16 (2006) and Figure 2 in which the benzene ring (R 1 ) of EDA, position 3 (R 3 ) and position 4 (R 4 ) can be further modified and have similar oxidation potential (Epa) and hydroxyl radical scavenging activity (IC 50 ).

[0038] EDA derivatives include 21 EDA derivatives confirmed in Bioorg. Med. Chem. Lett. (2015), which have the same anti-aggregation properties (http: / / dx.doi.org / 10.1016 / j.bmcl.2015.11.022).

[0039] EDA derivatives also include the compound of formula (I), especially the compound of formula (Ia) (BE) disclosed in Chinese Patent Application No. 201710036907.7. The content of this patent application document is incorporated herein by reference in its entirety.

[0040] 2. Dosage forms of edaravone

[0041] According to the needs of the administration route, edaravone and its salts or analogs or derivatives can be developed into various dosage forms. For example, it can be administered by intravenous drip, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or intravaginal routes.

[0042] Edaravone was first developed by Mitsubishi Tanabe Pharm Corp. (Osaka, Japan) for improving neurological symptoms, activities of daily living, and functional disorders caused by acute cerebral infarction. The marketed dosage form is an injection. Currently, edaravone has been widely used at home and abroad for the treatment of acute ischemic stroke (AIS) and amyotrophic lateral sclerosis (ALS), and also for the treatment of diseases related to excessive reactive oxygen species (ROS), such as cardiovascular diseases, etc.

[0043] CN105816423B discloses various preparations of edaravone (such as solid dispersion preparations) and their uses for effectively treating human oxidative stress-related diseases; CN 110996944A discloses a liquid aqueous solution of edaravone for treating oxidative stress-mediated neurodegenerative diseases in human patients, such as amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), cerebral amyloid angiopathy (CAA), Alzheimer's disease, and Parkinson's disease. CN 105616504B discloses edaravone or its analogs or derivatives for treating cerebral amyloid angiopathy (CAA) (the content of the above 3 patent application documents is incorporated herein by reference in its entirety).

[0044] Considering the convenience of administration and increasing the medication compliance of patients, the preferred dosage form is a solid preparation, preferably a solid dispersion preparation, for oral administration.

[0045] The solid dispersion preparation may include the active ingredient edaravone or its pharmaceutically acceptable salt or analog or derivative, a polymer carrier, and optionally a surfactant. The polymer carrier is selected from one or more of Soluplus, polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose acetate (HPMCAS), hydroxypropyl cellulose (HPC), and chitosan.

[0046] The surfactant includes anionic, cationic or amphoteric surfactants, and is selected from sodium dodecyl sulfonate, sodium dodecyl sulfate (SDS), sodium lauryl sulfate (SLS), polyoxyethylene sorbitan long-chain fatty acid esters, vitamin E-TPGS, bile salts, sodium deoxycholate, sodium glycocholate, polyoxyethylene polyoxypropylene glycol, and combinations thereof. Preferably, the surfactant is TPGS1000.

[0047] The preparation method of the oral administration preparation of the present invention can be prepared by referring to the conventional methods in the art, and it is particularly preferred to prepare the oral administration preparation of edaravone or its pharmaceutically acceptable salt by referring to the method disclosed in CN105816423B.

[0048] In the present invention, edaravone or its pharmaceutically acceptable salt or analog or derivative are sometimes collectively referred to as edaravone and used interchangeably.

[0049] According to the present invention, the administration dose of edaravone is between 0.1 mg / kg and 100 mg / kg. For treatment, the administration dose is preferably 0.1 - 50 mg / kg, such as 0.1 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 3.0 mg / kg, 5.0 mg / kg, 10.0 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg or 50 mg / kg, etc., and is administered twice a day to twice a week.

[0050] The dose and frequency of administration of the preparation of the present invention can vary according to the patient's condition, and sometimes it is necessary to administer at relatively short intervals and relatively high doses (for example, a dose of 5.0 - 35 mg / kg) until the progression of the disease is alleviated or terminated, preferably until the patient shows partial or complete improvement of the disease symptoms. The drugs of the present invention can be administered by parenteral, topical, intravenous, oral, subcutaneous, intra-arterial, intracranial, intrathecal, intraperitoneal, intranasal or intramuscular routes. Although other routes are equally effective, the typical route of administration is oral, followed by intramuscular injection. This type of injection is most commonly performed in the arm or leg muscles.

[0051] The present application provides the use of edaravone or a pharmaceutically acceptable salt, analogue or derivative thereof in the preparation of a drug for preventing and / or treating cognitive impairment caused by hypobaric hypoxia at high altitude.

[0052] When an animal body (including the human body) lives in a hypobaric hypoxia environment at high altitude for a long time, due to being in a hypoxic state for a long time, some unique trends of biochemical index changes will occur. These trends of index changes are different from those caused by usual intermittent hypoxia (hypoxia-reoxygenation-hypoxia-reoxygenation process), etc. For example, the level of superoxide dismutase (SOD) decreases significantly, and the ratio of reduced glutathione (glutathione, GSH) to oxidized glutathione (glutathione disulfide, GSSG), GSH / GSSG, also decreases significantly.

[0053] The present invention unexpectedly discovers that edaravone has very good preventive and therapeutic effects on cognitive impairment caused by hypobaric hypoxia at high altitude, especially has prominent therapeutic effects on cognitive dysfunction caused by chronic hypobaric hypoxia, and can improve biochemical indexes, hippocampal nerve defects, neuroinflammation, etc. caused by chronic hypobaric hypoxia.

[0054] To further understand the present invention, the preferred embodiments of the present invention will be described below in conjunction with examples. These descriptions are only examples to illustrate the features and advantages of the technical solutions of the present invention, rather than limiting the protection scope of the present invention.

[0055] Experimental purpose: To test the improvement effect of edaravone on cognitive dysfunction caused by hypobaric hypoxia at high altitude.

[0056] Experimental animals: All experimental procedures were carried out in accordance with the guiding principles of the Animal Ethics Committee of Army Medical University. Adult male C57BL / 6 mice (8-9 weeks old) were housed in a specific pathogen-free facility with a 12-hour light-dark cycle and had free access to standard feed and water.

[0057] Experimental design: To detect the effectiveness of edaravone as a preventive or therapeutic agent against hypobaric hypoxia, the mice were randomly divided into the following 4 groups, with 9 mice in each group:

[0058] Table 1: Experimental grouping design

[0059]

[0060] The edaravone oral solid dispersion granules used were prepared according to Example 7 of CN_105816423_B.

[0061] Test indicators

[0062] A series of behavioral tests were conducted on days 36-44, including:

[0063] 1. Open Field Test: The Open Field Test is used to evaluate general motor function. The apparatus is made of white plexiglass (40 cm × 40 cm × 40 cm). The mouse is placed in the central area and can freely explore the entire field. The total movement distance of the mouse within 10 minutes is recorded and analyzed.

[0064] 2. Novel Object Test: 24 hours after the adaptation phase (Open Field Test), during the acquisition phase, the mouse is allowed to explore two identical objects in the open field for 10 minutes. Two hours later, the mouse is allowed to explore a novel object and a familiar object in the open field for 5 minutes. The exploration times of the familiar object and the novel object are recorded. The discrimination rate is calculated as follows: 100% × novel object time / (novel object time + familiar object time).

[0065] 3. Barnes Maze: The maze consists of a white circular platform with a diameter of 91 cm and 20 holes with a diameter of 5 cm. A black escape box is placed under the target hole. The spatial cues around the maze and white noise (80 dB) can facilitate and motivate the mouse to find the target hole. Each time during the test, first, the mouse is placed in an opaque cylinder in the starting area for 10 seconds. Then, the cylinder is lifted and the white noise is turned on. When the mouse enters the target hole, the buzzer is turned off. During the adaptation phase (day 0), the mouse is allowed to explore the maze for 180 seconds and then stay in the box for 120 seconds. During the learning phase (days 1 - 4), the mouse is trained to find and enter the target hole within 180 seconds. After finding the target hole, all mice are allowed to stay in the box for 60 seconds. If the target hole is not found within 180 seconds, the mouse is guided to the target hole and allowed to stay in the hole for 60 seconds. Three trials are conducted each day, with a 15 - minute interval between each trial, for 4 consecutive days. During the test phase (day 5), the mouse explores the maze within 90 seconds. The latency of the mouse to enter the target hole and the number of errors in each trial are recorded and analyzed (for the schematic diagram, see Figure 1 ).

[0066] 4. Biochemical Assay: A total superoxide dismutase (SOD) detection kit (containing WST - 8) and a reduced glutathione (GSH) and oxidized glutathione (GSSG) detection kit (Beyotime) are used to evaluate the activity of SOD and the ratio of GSH / GSSG, respectively.

[0067] 5. Immunofluorescence staining and cell counting: After phenobarbital anesthesia, mice were perfused with 0.01 M phosphate buffered saline (PBS). Brains were dissected according to a previous protocol. Coronal brain sections (30 μm thick) were rinsed with 0.01 M PBS and then incubated with 0.3% Triton X-100 for 1 hour at room temperature. Then, primary antibody incubations were performed, including rabbit anti-ionized calcium-binding adapter molecule-1 (Iba1) (1:1000, Wako), mouse anti-glial fibrillary acidic protein (GFAP) (1:1000, Millipore), rabbit anti-sex determining region Y-box 2 (Sox2) (1:1000, Millipore), rabbit anti-doublecortin (DCX) (1:1000, Cell Signaling Technology), overnight at 4°C. After rinsing the sections with 0.01 M PBS, they were incubated with fluorescently labeled secondary antibodies (including Cy3 (1:500, donkey anti-rabbit, Jackson ImmunoResearch), Alexa 488 (1:500, donkey anti-mouse, Jackson ImmunoResearch)) for 2 hours at 37°C. Finally, nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich). After staining, photographs were taken using a Zeiss microscope.

[0068] According to the previous method (Zhang, R., Cai, Y., Xiao, R., Zhong, H., Li, X., Guo, L., Xu, H., and Fan, X. (2019). Human amniotic epithelial cell transplantation promotes neurogenesis and ameliorates social deficits in BTBR mice. Stem Cell Res Ther 10, 153. 10.1186 / s13287-019-1267-0), radial glial cells were labeled by GFAP+Sox2+, and their extended apices passed through the granular cell layer. Five matched sections across the dentate gyrus (DG) region were counted using a blind method to evaluate the number of Sox2+ / GFAP+ and DCX+ cells in the DG.

[0069] Statistical analysis

[0070] Statistical analysis was performed using SPSS (version 25.0 for Windows, IBM). Data with a normal distribution were expressed as mean ± SEM. One-way analysis of variance and Turkey's post hoc test were used for multiple comparisons. For the evaluation data of repeated time points, two-way analysis of variance was performed, followed by Turkey's post hoc test.

[0071] Results:

[0072] 1. Results of the open-field test

[0073] The open-field test was used to evaluate general motor function. Compared with mice raised under normal conditions, long-term low-pressure hypoxia significantly impaired the spontaneous motor function of mice in the high-altitude + placebo group, as manifested by a decrease in the total movement distance of mice in the open-field test (P < 0.001). Compared with the high-altitude + placebo group, administration of edaravone significantly increased the total movement distance of mice in the high-altitude + treatment group (P < 0.05) or the high-altitude + prevention group (P < 0.01) (Table 2, Figure 2 , 3).

[0074] Table 2: Total movement distance of mice in the open-field test (mean ± standard deviation)

[0075] Total movement distance (cm) Plain + placebo <![CDATA[3443.49±488.11 ### > High altitude + placebo 2412.83±244.02*** High altitude + treatment <![CDATA[2986.13±384.41 # > High altitude + prevention <![CDATA[3027.01±317.44 ## >

[0076] Note: Compared with the plain + placebo group, ***P < 0.001; compared with the high-altitude + placebo group, # P < 0.05, ## P < 0.01, ### P < 0.001. n = 9 / group. One-way analysis of variance and Turkey's post hoc test.

[0077] 2. Results of the novel object recognition experiment

[0078] The novel object recognition experiment was used to evaluate episodic declarative memory. In the novel object recognition experiment, the discrimination rate of mice in the high altitude + placebo group was lower than that of mice in the plain + placebo group (P < 0.01), while edaravone administered for 5 weeks could significantly reverse the memory deficit of mice (P < 0.01)( Figure 3 , 4). In addition, compared with mice in the high altitude + placebo group, the edaravone treatment group also showed a trend of higher discrimination rate (P = 0.079) (Table 3, Figure 4 , 5).

[0079] Table 3: Discrimination rate of mice in the novel object recognition experiment (mean ± standard deviation)

[0080] Discrimination rate (%) Plain + placebo <![CDATA[59.12±10.07 ## > High altitude + placebo 34.34±10.15** High altitude + treatment 48.45±16.86 High altitude + prevention <![CDATA[58.92±9.15 ## >

[0081] Note: Compared with the plain + placebo group, **P < 0.01; compared with the high altitude + placebo group,

[0082] ## P < 0.01. n = 9 / group. One-way ANOVA and Turkey post hoc test.

[0083] 3. Results of the Barnes maze experiment

[0084] The Barnes maze was used to evaluate learning and memory functions. In the learning phase of the Barnes maze, compared with the plain + placebo group, the latency (P < 0.01) and the number of errors (P < 0.001) of mice in the high altitude + placebo group to enter the target hole were significantly increased. The performance of mice in the high altitude + treatment or high altitude + prevention groups in terms of latency and the number of errors during the learning phase was much better than that of mice in the high altitude + placebo group (P < 0.05 for all comparisons) (Table 4, 5; Figure 6 , 7).

[0085] Table 4: Latency of mice to enter the target hole in the Barnes maze experiment (mean ± standard deviation)

[0086] <![CDATA[Plain + Placebo ## > High altitude + placebo** <![CDATA[High altitude + treatment # > <![CDATA[Plateau + Prevention # > Day 1 138.70±39.39 152.51±31.80 142.96±38.13 132.69±33.80 Day 2 68.55±30.56 115.41±32.00 67.16±18.02 67.32±42.47 Day 3 43.21±21.50 64.48±32.42 44.15±23.40 40.38±15.07 Day 4 29.39±23.21 81.41±49.61 48.18±27.96 45.47±27.19

[0087] Note: Compared with the plain + placebo group, **P < 0.01; compared with the high altitude + placebo group, # P < 0.05, ## P < 0.01. n = 9 / group. Two-way ANOVA was used and Turkey post hoc test was performed.

[0088] Table 5: Errors made by mice during the learning period in the Barnes maze experiment

[0089] <![CDATA[Plain + placebo ### > High altitude + placebo*** <![CDATA[High altitude + treatment # > <![CDATA[Plateau + Prevention # > Day 1 45.78±19.38 51.67±16.05 64.30±14.65 55.56±18.61 Day 2 20.96±9.74 41.04±16.15 20.74±9.51 25.00±11.62 Day 3 15.33±8.42 29.59±12.95 17.74±8.34 16.96±8.50 Day 4 8.93±12.26 25.37±18.58 13.48±9.18 16.81±13.88

[0090] Note: Compared with the plain + placebo group, ***P < 0.001; compared with the high altitude + placebo group, # P < 0.05, ### P < 0.001. n = 9 / group. Two-way ANOVA was used, followed by Tukey's post hoc test.

[0091] 4. Results of biochemical tests

[0092] Compared with the mice in the plain + placebo group, the SOD (P < 0.01) and GSH / GSSG (P < 0.001) in the brains of the mice in the high altitude + placebo group decreased significantly. Edaravone treatment and intervention could partially reverse the changes in SOD (high altitude + treatment vs. high altitude + placebo, P < 0.05; high altitude + prevention vs. high altitude + placebo, P < 0.01) and GSH / GSSG (high altitude + treatment vs. high altitude + placebo, P < 0.001; high altitude + prevention vs. high altitude + placebo, P < 0.001) (Table 6, Figure 8 A - B). It is suggested that edaravone can significantly improve the biochemical indexes caused by hypobaric hypoxia.

[0093] Table 6: Biochemical levels of mice (mean ± standard deviation)

[0094] SOD GSH / GSSG Plain + placebo <![CDATA[16.29±2.47 ## > <![CDATA[17.43±3.69 ### > High altitude + placebo 8.51±3.92** 8.01±1.07*** High altitude + treatment <![CDATA[13.99±3.16 # > <![CDATA[14.13±1.66 ### > High altitude + prevention <![CDATA[15.34±4.88 ## > <![CDATA[14.34±2.70 ### >

[0095] Note: Compared with the plain + placebo group, **P < 0.01, ***P < 0.001; compared with the high altitude + placebo group, # P < 0.05, ## P < 0.01, ### P < 0.001. n = 8 / group. One-way ANOVA and Tukey's post hoc test were used.

[0096] 5. Results of detection of neuroinflammatory levels

[0097] The results of immunofluorescence staining showed that compared with the mice in the plain + placebo group, the number of microglia (Iba1+) in the DG region of the mice in the high altitude + placebo group increased significantly (P < 0.001), indicating that hypobaric hypoxia increased the inflammatory level of the hippocampus in mice. In addition, compared with the high altitude + placebo group, the number of Iba1+ cells in the DG of the mice in the high altitude + treatment group (P < 0.01) and the high altitude + prevention group (P < 0.001) was less (Table 7, Figure 9 ). It is suggested that edaravone can reduce the neuroinflammatory level caused by hypobaric hypoxia.

[0098] Table 7: Neuroinflammatory levels of mice (mean ± standard deviation)

[0099] Number of Iba1+ cells / DG Plain + placebo <![CDATA[21.13±1.22 ### > High altitude + placebo 26.76±1.77*** High altitude + treatment <![CDATA[23.80±0.97**, ## > High altitude + prevention <![CDATA[22.09±1.95 ### >

[0100] Note: Compared with the plain + placebo group, **P < 0.01,***P < 0.001; compared with the high altitude + placebo group, ## P < 0.01, ### P < 0.001. n = 9 / group. One-way ANOVA and Turkey post hoc test.

[0101] 6. Hippocampal neurogenesis disorder

[0102] Chronic hypobaric hypoxia leads to hippocampal neurogenesis disorder in mice, manifested as a decrease in the number of neural progenitor cells (NPCs) (Sox2+ / GFAP+) in the DG region of mice in the high altitude + placebo group compared with those in the plain + placebo group (P < 0.001). Compared with the high altitude + placebo group, edaravone administration can significantly increase the number of Sox2+ / GFAP+ cells in the DG of the high altitude + treatment group (P < 0.01) and the high altitude + prevention group (P < 0.01) (Table 8, Figure 10 ).

[0103] Table 8: Number of (Sox2+ / GFAP+) in the DG region of the mouse hippocampus (mean ± standard deviation)

[0104] Number of Sox2+GFAP+ cells / DG Plain + placebo <![CDATA[25.00±3.87 ### > High altitude + placebo 16.67±3.84*** High altitude + treatment <![CDATA[22.67±3.35 ## > High altitude + prevention <![CDATA[23.44±2.70 ## >

[0105] Note: Compared with the plain + placebo group, ***P < 0.001; compared with the high altitude + placebo group, ## P < 0.01, ### P < 0.001. n = 9 / group. One-way ANOVA and Turkey post hoc test.

[0106] Compared with the plain + placebo group, the number of DCX+ cells in the DG region of mice in the high altitude + placebo group (P < 0.001), high altitude + treatment group (P < 0.05), or high altitude + prevention group (P < 0.05) increased significantly, indicating that long-term hypobaric hypoxia increases the differentiation of NPCs in the hippocampus. The number of DCX+ cells in the high altitude + treatment group (P < 0.05) and the high altitude + prevention group (P < 0.05) was significantly lower than that in mice in the high altitude + placebo group (Table 9, Figure 11 ).

[0107] Table 9: Number of DCX+ cells in the DG region of the mouse hippocampus (mean ± standard deviation)

[0108] Number of DCX+ cells / DG Plain + placebo <![CDATA[45.71±6.05 ### > High altitude + placebo 61.80±4.20*** High altitude + treatment <![CDATA[53.49±6.31 # > High altitude + prevention <![CDATA[54.13±5.23 # >

[0109] Note: Compared with the plain + placebo group, ***P < 0.001; compared with the high altitude + placebo group, # P < 0.05, ###P < 0.001. n = 9 / group. One-way ANOVA and Turkey post hoc test.

[0110] It is indicated that edaravone can improve the hippocampal neurogenesis disorder caused by hypobaric hypoxia.

[0111] In summary, edaravone has a significant protective effect in improving hippocampal nerve defects, neuroinflammation and cognitive dysfunction caused by chronic hypobaric hypoxia, can significantly improve its biochemical indexes, and can even partially reverse the changes of typical biochemical indexes such as SOD and GSH / GSSG ratio.

[0112] The description of the above specific embodiments is only used to help understand the core idea of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the technical solution of the present invention, but these improvements and modifications also fall within the scope of the claims of the present invention.

Claims

1. Use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof in the preparation of a drug for preventing and / or treating cognitive impairment caused by hypobaric hypoxia at high altitude.

2. The application according to claim 1, wherein, The drug comprises edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof and one or more pharmaceutically acceptable excipients.

3. The application according to claim 2, wherein, The administration route of the drug is selected from intravenous drip, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or intravaginal administration, and oral administration is preferred.

4. The application according to claim 1, wherein The dosage form of the drug is a solid preparation, and a solid dispersion preparation is preferred.

5. The application according to claim 4, wherein, The solid dispersion preparation comprises the active ingredient edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof, a polymer carrier, and an optional surfactant.

6. The application according to claim 5, wherein, The polymer carrier is selected from one or more of Soluplus, polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose acetate (HPMCAS), hydroxypropyl cellulose (HPC), and chitosan.

7. The application according to claim 5, wherein The surfactant includes anionic, cationic or amphoteric surfactants, and is selected from sodium dodecyl sulfonate, sodium dodecyl sulfate (SDS), sodium lauryl sulfate (SLS), polyoxyethylene sorbitan long-chain fatty acid esters, vitamin E-TPGS, bile salts, sodium deoxycholate, sodium glycocholate, polyoxyethylene polyoxypropylene glycol, and combinations thereof.

8. The application according to claim 5, wherein The surfactant is TPGS1000.

9. The application according to claim 1, wherein, The unit dose of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof is 0.001 - 1000 mg, preferably 0.1 - 100 mg, more preferably 1 - 50 mg.

10. The application according to claim 1, wherein, The administration dose of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof is 0.1 - 100 mg / kg, preferably 1 - 24 mg / kg.

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