Use of edaravone in prevention and treatment of acute high-altitude illnesses

Abstract

Provided in the present disclosure is the use of edaravone and a pharmaceutically acceptable salt thereof in the prevention and / or treatment of acute high-altitude illnesses caused by acute high-altitude hypobaric hypoxia. The acute high-altitude illnesses include, for example, acute mountain sickness (AMS), high-altitude cerebral edema (HACE), and high-altitude pulmonary edema (HAPE). Additionally provided in the present disclosure is the use of edaravone and a pharmaceutically acceptable salt thereof in the prevention and / or treatment of high-altitude pulmonary hypertension (HAPH) and inflammatory responses caused by acute high-altitude hypobaric hypoxia.

Description

Application of edaravone in the prevention and treatment of acute high-altitude illness Technical Field

[0001] This disclosure pertains to the pharmaceutical field and specifically relates to a novel use of edaravone, particularly in the preparation of edaravone for the prevention and / or treatment of diseases caused by high altitudes. Background Technology

[0002] The Qinghai-Tibet Plateau, known as the "Roof of the World," is located in the southwestern border region of my country, with an average altitude of over 4,000 meters. It plays a crucial role in stabilizing the climate system of my country and even East Asia, and serves as an important ecological security barrier for my country and the world. According to the "2023 China Statistical Yearbook," the population of the Qinghai-Tibet Plateau at the end of 2022 was 9.59 million, and statistics from the Ministry of Culture and Tourism of the People's Republic of China show that the number of tourists residing on the Qinghai-Tibet Plateau reached 99.9332 million in 2023. According to the altitude classification standards jointly established by the International Organization for Standardization (ISO) and the International Civil Aviation Organization (ICAO), the Qinghai-Tibet Plateau belongs to the high-altitude and extremely high-altitude regions. The harsh environment of low pressure and oxygen deficiency, coupled with increasingly severe climate change, has become a major obstacle to human survival and development in this region.

[0003] Hypoxia is a pathological process in which insufficient oxygen supply or impaired oxygen utilization in tissues leads to abnormal changes in their metabolism, function, and morphology. Long-term and severe hypoxia can cause failure of vital organs such as the heart and brain, ultimately leading to death.

[0004] At altitudes above 3500m, the low pressure and low oxygen environment can cause altitude sickness, including chest tightness, shortness of breath, nausea, vomiting, headache, and palpitations, which can be life-threatening in severe cases. Therefore, improving one's tolerance to hypoxia is crucial for adapting to life at high altitudes.

[0005] With the increasing demand for high-altitude tourism, sports competitions, urban construction, and military activities, more and more people are traveling from plains to high-altitude areas. Research on the prevention and treatment of bodily damage caused by the low-oxygen environment of high-altitude areas has become a hot topic.

[0006] Currently, there are few medications available for treating altitude sickness, with acetazolamide being the only FDA-approved drug for preventing and treating altitude sickness. The Wilderness Medical Society has reported the use of acetazolamide and dexamethasone to treat altitude sickness symptoms and for the prevention and treatment of acute mountain sickness (AMS) and high-altitude acute brain injury (HACE) (Wilderness Medical Society Clinical Practice Guidelines for the Prevention, Diagnosis, and Treatment of Acute Altitude Illness: 2024 Update). However, the effective dosage of acetazolamide reported in the literature is inconsistent, and both acetazolamide and dexamethasone have significant toxic and side effects, limiting their use in routine prevention.

[0007] On the other hand, traditional Chinese medicine compound preparations, mainly containing plateau plants such as Rhodiola rosea, can also be used to alleviate high-pressure reactions, but they face problems such as relatively limited resources, high prices, and over-exploitation damaging the plateau's ecological environment.

[0008] There is a need in this field for effective drugs to treat acute hypobaric hypoxia, which is of great significance to the health and safety of people exposed to hypoxia at high altitudes. Summary of the Invention

[0009] The inventors discovered that when animals were given edaravone prophylactically and then rapidly advanced to high altitudes, their blood oxygen saturation was significantly higher than that of animals that were not given prophylactic medication. Lung tissue damage was alleviated to varying degrees, and the animals showed a protective effect against edema in organs such as the brain and lungs caused by rapid advancement to high altitudes, reducing the inflammatory response.

[0010] The purpose of this disclosure is to provide a novel use of edaravone for the prevention and / or treatment of acute altitude sickness, including acute altitude sickness (AMS), high-altitude encephalopathy (HACE), and high-altitude angina (HAPE).

[0011] First, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof (preferably alone as the sole active ingredient) in the preparation of a medicament for the prevention and / or treatment of acute high-altitude illnesses.

[0012] The acute altitude sickness is at least one selected from the group consisting of: acute mountain sickness (AMS), high-altitude acute brain injury (HACE), and high-altitude acute lung injury (HAPE). The AMS includes mild AMS, moderate-to-severe AMS, or altitude sickness symptoms expressed as a Lake Louise score of 3-12. In one embodiment, the acute altitude sickness is selected from HACE and HAPE.

[0013] On the other hand, this disclosure also provides a pharmaceutical composition for the prevention and / or treatment of acute high-altitude illnesses, comprising edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof, and one or more pharmaceutically acceptable excipients.

[0014] According to this disclosure, the pharmaceutical composition can be administered via intravenous infusion, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or vaginal routes. Preferably, the pharmaceutical composition is administered orally.

[0015] In one embodiment of this disclosure, the oral dosage form is a solid dosage form, preferably a solid dispersion, comprising the active ingredient edaravone or a pharmaceutically acceptable salt thereof, a polymer carrier selected from Soluplus, polyethylene glycol (PEG), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose acetate (HPMCAS), hydroxypropyl cellulose (HPC), and chitosan (preferably Soluplus and / or HPMC, more preferably Soluplus), and optionally a surfactant such as TPGS1000. 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, for example 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.

[0016] This disclosure relates to a method for preventing and / or treating acute altitude sickness (AMS) with edaravone or a pharmaceutically acceptable salt, analog, or derivative thereof, comprising administering a therapeutically effective amount of edaravone or a pharmaceutically acceptable salt, analog, or derivative thereof to a subject in need. Acute altitude sickness includes AMS, HACE, and HAPE, among others.

[0017] This disclosure also relates to edaravone or a pharmaceutically acceptable salt or analog or derivative thereof, for the prevention and / or treatment of acute high-altitude illnesses, the medicament comprising edaravone or a pharmaceutically acceptable salt or analog or derivative thereof.

[0018] This publication unexpectedly discovered that edaravone has very good preventive and therapeutic effects on acute high-altitude diseases, especially on acute high-altitude brain injury and acute high-altitude lung injury caused by acute high-altitude hypoxia, and can improve brain injury, blood oxygen concentration, degree of lung tissue damage, and degree of pulmonary edema.

[0019] In addition, this disclosure provides the use of edaravone and its pharmaceutically acceptable salts in the prevention and / or treatment of inflammatory responses caused by high-altitude pulmonary hypertension (HAPH) and acute high-altitude hypobaric hypoxia. Attached Figure Description

[0020] To more clearly describe the technical solutions of this disclosure, a brief introduction will be given below in conjunction with the accompanying drawings. Obviously, these drawings are merely some specific embodiments described in this disclosure. This disclosure includes, but is not limited to, these drawings.

[0021] Figure 1 shows the blood gas analysis results of rats exposed to hypoxia after receiving different doses of EDA. Figure 1A shows the partial pressure of oxygen (PaO2) in the blood of rats exposed to hypoxia after receiving different doses of EDA. Symbols in the figure: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ## indicates P < 0.01, ### indicates P < 0.001; compared with group E, △△ indicates P < 0.01. Figure 1B shows the blood pH of the rats.

[0022] Figure 2 shows the wet-dry ratio of lung tissue in rats exposed to hypobaric hypoxia after receiving different doses of EDA. Symbols in the figure: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ## indicates P < 0.01, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05.

[0023] Figure 3 shows the hypoxia indices in the lung tissue of rats exposed to low-pressure hypoxia after receiving different doses of EDA. Symbols in the figure: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.01; compared with group B, # indicates P < 0.05, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05, △△△ indicates P < 0.001.

[0024] Figure 4 shows the hypoxia indices in the lung tissue of rats exposed to low-pressure hypoxia after receiving different doses of EDA. Symbols in the figure: Compared with group A, * indicates P < 0.05, *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ### indicates P < 0.001; compared with group E, △ indicates P < 0.01, △△ indicates P < 0.01.

[0025] Figure 5 shows the hypoxia indices of rat brain tissue exposed to low-pressure hypoxia after administration of different doses of EDA. Symbols in the figure: Compared with group A, *** indicates P<0.001, ** indicates P<0.01, and * indicates P<0.05; compared with group B, ### indicates P<0.001, ## indicates P<0.01, and # indicates P<0.05; compared with group E, △ indicates P<0.05, and △△△ indicates P<0.001.

[0026] Figure 6 shows the hypoxia indices of rat brain tissue exposed to low-pressure hypoxia after administration of different doses of EDA. Symbols in the figure: Compared with group A, * indicates P<0.05, *** indicates P<0.001; compared with group B, ## indicates P<0.01, ### indicates P<0.001; compared with group E, △ indicates P<0.05.

[0027] Figure 7 shows the hypoxia indices in the serum of rats exposed to low-pressure hypoxia after receiving different doses of EDA. Symbols in the figure: *** indicates P < 0.001 compared to group A; ### indicates P < 0.001 compared to group B; # indicates P < 0.05.

[0028] Figure 8 shows the hypoxia indices in the serum of rats exposed to low-pressure hypoxia after receiving different doses of EDA. Symbols in the figure: *** indicates P < 0.001 compared to group A; ### indicates P < 0.001 compared to group B; △ indicates P < 0.05 compared to group E.

[0029] Figure 9 shows HE-stained sections of rat lung tissue exposed to low-pressure hypoxia after receiving different doses of EDA.

[0030] Figure 10 shows the protein concentration in the bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia after receiving different doses of EDA. Symbols in the figure: *** indicates P < 0.001 compared to group A; ### indicates P < 0.001 compared to group B; △ indicates P < 0.05 compared to group E.

[0031] Figure 11 shows cell counts in bronchoalveolar lavage fluid from rats exposed to hypobaric hypoxia after receiving different doses of EDA. Symbols in the figure: Compared with group A, *** indicates P < 0.001, * indicates P < 0.05; compared with group B, ### indicates P < 0.001, # indicates P < 0.05; compared with group E, △△ indicates P < 0.01.

[0032] Invention Details

[0033] definition

[0034] High-altitude illnesses refer to diseases caused by the inability of the human body to adapt to the high-altitude environment when entering or moving from a high-altitude area. They can be divided into acute high-altitude illnesses (HAIs) and chronic high-altitude illnesses (CMS).

[0035] Acute high-altitude illnesses include acute mountain sickness (AMS), high-altitude acute brain injury (HACE), and high-altitude acute lung injury (HAPE). Chronic high-altitude illnesses are characterized by excessive erythrocyte proliferation and significant pulmonary hypertension, with a prolonged course and eventual right heart failure.

[0036] Acute mountain sickness (AMS) is a variety of pathological reactions that occur when the body is exposed to a low-oxygen environment for a short period of time after entering a high-altitude area.

[0037] AMS diagnosis is primarily based on symptoms. Symptoms of AMS are atypical and mainly include headache, loss of appetite, nausea, fatigue, dizziness or vertigo, which usually appear within hours to three days after arriving at high altitudes.

[0038] One of the reference criteria for diagnosing AMS is the Lake Louise Acute Mountain Sickness Score. The Lake Louise score includes scores for four main symptoms: headache, gastrointestinal symptoms, fatigue or weakness, and dizziness or vertigo. 0 indicates no discomfort, 1 indicates mild, 2 indicates moderate, and 3 indicates severe symptoms. A Lake Louise score of 3-5 indicates mild AMS, 6-9 indicates moderate AMS, and 10-12 indicates severe AMS.

[0039] Table 1 AMS Classification

[0040] Further progression of acute myocardial infarction (AMS) can lead to more severe high-altitude acute brain injury (HACE) and high-altitude acute lung injury (HAPE). Reports suggest that disruption of the blood-brain barrier causes HACE, while increased pulmonary capillary pressure and associated pressure failure trigger HAPE.

[0041] HACE (high altitude cerebral edema) is diagnosed primarily based on symptoms. Ataxia is usually the earliest clinical manifestation of HACE, and other symptoms include headache, nausea, vomiting, apathy, irritability, fatigue, and altered consciousness. In areas with better medical facilities, cranial imaging and laboratory tests can rule out other possible diagnoses, such as cerebrovascular accident, carbon monoxide poisoning, and traumatic brain injury.

[0042] When HACE occurs, cranial MRI typically shows high signal intensity in the corpus callosum on fluid attenuated inversion recovery and T2 sequences; at the same time, microbleeds consistent with blood-brain barrier disruption can also be observed on susceptibility-weighted imaging, mainly in the corpus callosum region.

[0043] HAPE (high altitude pulmonary edema) is a type of high-flow-rate, high-pressure, high-osmolar edema, a life-threatening non-cardiac pulmonary edema.

[0044] Patients with high-altitude pulmonary edema (HAPE) may experience symptoms such as dry cough, fatigue, weakness, chest tightness, and shortness of breath. As the condition progresses, patients may experience difficulty breathing even with slight activity or at rest, and may also develop cyanosis and cough up pink frothy sputum. The bronchoalveolar lavage fluid of patients with high-altitude pulmonary edema has significantly higher protein levels than the control group and contains a large number of red blood cells, white blood cells, and macrophages. Autopsy revealed a significant increase in protein levels in the pulmonary edema fluid; scattered alveolar hemorrhages; and fibrin exudation and hyaline membrane formation in the blood vessel walls. Some reports suggest that HAPE is primarily caused by elevated pulmonary artery pressure, with inflammatory responses being a consequence, which in turn further exacerbates HAPE.

[0045] Acute high-altitude hypoxia: In this disclosure, it means hypoxia within 1-7 days or several weeks after arriving at a plateau or high-altitude area.

[0046] Inflammatory response caused by acute high-altitude hypoxia: In this disclosure, the inflammatory response includes inflammatory responses involved in pulmonary edema and cerebral edema in HACE and HAPE.

[0047] PAH (pulmonary hypertension) is hemodynamically defined as a mean pulmonary artery pressure ≥25 mmHg at rest and a normal left ventricular filling pressure ≥30 mmHg during exercise. PAH is caused by uncontrolled pulmonary vascular remodeling, leading to increased pulmonary vascular resistance. High pulmonary vascular resistance results in progressively increasing pulmonary artery pressure and persistent vasoconstriction, leading to poorly adaptive right ventricular hypertrophy. PAH is a fundamental factor in the development of high-altitude pulmonary edema. Patients with high-altitude pulmonary edema, whether in the early stages or during the recovery phase, have significantly higher pulmonary artery pressure than normal individuals at the same altitude. As altitude increases, atmospheric pressure decreases, leading to a decrease in PiO2, PaO2, and SaO2%, resulting in hypoxemia. This causes hypoxic vasoconstriction in the pulmonary vessels, increasing PVR (pulmonary vascular resistance), and PAP (prostatic acid phosphatase) levels with altitude.

[0048] High-altitude pulmonary hypertension (HAPH) is a pathological syndrome caused by hypoxia, characterized by pulmonary vasoconstriction and pulmonary vascular remodeling. It is a chronic high-altitude disease with high morbidity and mortality rates in high-altitude regions. Studies indicate that long-term hypoxia can promote HAPH development through vascular remodeling (a major factor) and inflammatory responses. The pathological features of HAPH mainly include: 1) endothelial cell damage and proliferation; 2) proliferation and phenotypic transformation of vascular smooth muscle cells; 3) myomodulation of non-muscular arterioles; and 4) adventitia fibroblast proliferation and extracellular matrix deposition.

[0049] HH: Hypobaric hypoxia. At high altitudes, the oxygen concentration remains relatively constant under atmospheric pressure, but as atmospheric pressure decreases, the partial pressure of oxygen (PIO2) in the inhaled gas decreases proportionally, resulting in hypobaric hypoxia. In this disclosure, "hypobaric hypoxia" is used with the same meaning as "hypobaric hypoxia" and "low-pressure hypoxia".

[0050] PaO2: Arterial partial pressure of oxygen (PaO2)

[0051] PIO2: Partial pressure of oxygen in inhaled gas, calculated as PIO2 = FiO2 × (P B -P H2O Where FiO2 is the inhaled oxygen concentration, P H2O This refers to the airway water vapor pressure, typically 47 mmHg; P B It is 1 standard atmosphere.

[0052] Acetazolamide (ACZ) is a carbonic anhydrase (CA) inhibitor that increases bicarbonate excretion, promotes diuresis, stimulates the respiratory tract, increases arterial oxygen partial pressure, reduces cerebrospinal fluid production, and promotes ion transport across the blood-brain barrier. ACZ is recommended for use in the Wilderness Medical Society Clinical Practice Guidelines for the Prevention, Diagnosis, and Treatment of Acute Altitude Illness: 2024 Update.

[0053] The term “pharmaceutical acceptable” means approved by regulatory authorities such as the EMEA (Europe) and / or the FDA (US) and / or any other national regulatory authority for use in animals, preferably in humans.

[0054] The term "excipient" refers to a diluent, adjuvant, or carrier that is administered in conjunction with a therapeutic agent. Examples of appropriate pharmaceutical excipients are described in "Remington's Pharmaceutical Sciences" by EW Martin.

[0055] Soluplus: A polyethylene oxide-polyethylene polymer copolymer with the molecular formula (C8H13NO·C4H6O2·C2H4O)x and the chemical formula (PEO)m(PPO)n(PEO)m, where PEO represents polyethylene oxide and PPO represents polypropylene ether. Also known as PCL-PVAc-PE. Soluplus is a solvent-based solubilizer.

[0056] compound

[0057] 1. Edaravone and its salts or analogues or derivatives

[0058] Edaravone (EDA) is a pyrazolinone derivative with the chemical name 3-methyl-1-phenyl-2-pyrazolin-5-one and the molecular formula C2. 10 H 10 N2O, the structural formula is shown below:

[0059] The core structure or active center of EDA is 2-pyrazolin-5-one, which plays a major role in free radical scavenging activity.

[0060] EDA is a nitrogen-containing heterocyclic compound that is basic and readily reacts with acids to form salts. Its pharmaceutically acceptable salts include inorganic and organic acid salts commonly found in the field. Inorganic acid salts include hydrochlorides, carbonates, phosphates, etc., while organic acid salts include citrates, tartrates, acetates, oxalates, salicylates, malates, lactates, etc.

[0061] EDA analogs include a methyl group at the 3-position of the pyrazoline ring, which can be ethyl, propyl, or other lower (C) groups. 1-6The phenyl group may be alkyl or lower alkoxy, such as methoxy, ethoxy, etc.; or the methyl group at position 3 may be H, while the H at position 4 may be replaced by a lower alkyl or alkoxy group. Edaravone derivatives include esters, where the ketone at position 5 of the pyrazoline ring is converted to an enol, which reacts with a carboxylic acid to form an ester, such as a methyl ester, ethyl ester, etc. The ester (precursor) is then hydrolyzed in vivo and converted back to a ketone. Additionally, the phenyl group may optionally be substituted with one or more substituents selected from lower alkyl, lower alkoxy, nitro, halogen, etc.

[0062] EDA derivatives include up to 18 EDA derivatives disclosed in Tables 1-2 and Figure 2 of Bioorg. Med. Chem. Lett. 16 (2006), of which the benzene ring (R) of EDA is... 1 ), Position 3 (R) 3 ) and position 4 (R 4 It can be further modified to possess similar oxidation potential (Epa) and hydroxyl radical scavenging activity (IC). 50 ).

[0063] EDA derivatives include 21 EDA derivatives identified in Bioorg. Med. Chem. Lett. (2015) that have the same anti-aggregation properties (http: / / dx.doi.org / 10..1016 / j.bmcl.2015.11.022).

[0064] EDA derivatives also include compounds of formula (I) disclosed in Chinese Patent Application No. 201710036907.7, especially compound (BE) of formula (Ia). The entire contents of that patent application are incorporated herein by reference.

[0065] 2. Dosage forms of edaravone

[0066] Edaravone and its salts or analogues or derivatives can be developed into various dosage forms depending on the route of administration required, such as intravenous infusion, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or vaginal administration.

[0067] Edaravone was first developed by Mitsubishi Tanabe Pharm Corp. (Osaka, Japan) to improve neurological symptoms, activities of daily living, and functional impairments caused by acute ischemic stroke. It is marketed as an injectable formulation. Currently, edaravone is widely used both domestically and internationally to treat acute ischemic stroke (AIS) and amyotrophic lateral sclerosis (ALS), and also for treating diseases related to excessive reactive oxygen species (ROS), such as cardiovascular diseases.

[0068] CN105816423B discloses various formulations of edaravone (e.g., solid-phase dispersions) and their use in treating human oxidative stress-related diseases; CN 110996944A discloses a liquid aqueous solution of edaravone for treating human patients with oxidative stress-mediated neurodegenerative diseases, 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 analogues or derivatives for treating cerebral amyloid angiopathy (CAA) (the contents of the above three patent applications are incorporated herein by reference in their entirety).

[0069] Considering the convenience of administration and to increase patient compliance, the preferred dosage form is a solid dosage form, preferably a solid dispersion, administered orally.

[0070] Solid dispersion formulations may include the active ingredient edaravone or its pharmaceutically acceptable salts or analogues or derivatives, 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.

[0071] 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.

[0072] The method for preparing the oral formulation disclosed herein can be obtained by referring to conventional methods in the art, and is particularly preferred for preparing oral formulations of edaravone or its pharmaceutically acceptable salts by referring to the method disclosed in CN105816423B.

[0073] In this disclosure, edaravone or its pharmaceutically acceptable salts or analogs or derivatives are sometimes collectively referred to as edaravone and used interchangeably.

[0074] According to this disclosure, the dosage of edaravone is between 0.1 mg / kg and 100 mg / kg. For therapeutic purposes, the preferred dosage is 0.1-50 mg / kg, for example, 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, administered twice daily to twice weekly.

[0075] When used for the prevention of altitude sickness, it is recommended that subjects begin using this formulation 2 days or 1 day before commencing their mountaineering activity from an altitude below 1200 meters, and continue using it. An optional dose is 1-2 times daily. When used to treat altitude sickness, this formulation should be started when the subject's altitude rapidly increases and / or symptoms of Alzheimer's disease (AMS), Hepatocellular carcinoma (HACE), or Hyperalgesia-Hyperalgesia (HAPE) appear, and continued until the symptoms partially or completely improve. An optional dose is once every 12 hours or once every 24 hours, adjusted according to the symptoms.

[0076] The use of this disclosed formulation may be combined with the following measures: stopping the subject's ascent, reducing the subject's altitude, administering oxygen to the subject, or using a portable hyperbaric oxygen chamber.

[0077] The dosage and frequency of administration of the formulation disclosed herein may vary depending on the patient’s condition, and may sometimes require relatively short intervals and relatively high doses (e.g., 5.0-35 mg / kg) until the progression of the disease is reduced or terminated, preferably until the patient shows partial or complete improvement in the symptoms of the disease.

[0078] The disclosed drug can be administered via parenteral, local, 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 administered in the muscles of the arm or leg.

[0079] use

[0080] This disclosure provides for the use of edaravone or its pharmaceutically acceptable salts or analogs or derivatives in the preparation of medicaments for the prevention and / or treatment of acute high-altitude illnesses.

[0081] In one embodiment, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof in the preparation of a medicament for the prevention and / or treatment of acute high-altitude diseases caused by acute high-altitude hypoxia, including acute altitude sickness (AMS), high-altitude acute brain injury (HACE), and high-altitude acute lung injury (HAPE).

[0082] In one embodiment, the AMS of this disclosure includes mild AMS, moderate to severe AMS, or altitude sickness symptoms expressed as a Lake Louise score of 3-12. Those skilled in the art will understand that the AMS of this disclosure does not include further aggravated symptoms.

[0083] In one embodiment, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof for the prevention and / or treatment of acute myasthenia gravis (AMS) and / or for reducing the Lake Louise score of a subject. Symptoms of AMS include, but are not limited to, headache, loss of appetite, nausea, fatigue, dizziness or vertigo, decreased PiO2, and hypoxemia.

[0084] In one embodiment, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof for the prevention and / or treatment of HACE. Symptoms of HACE include, but are not limited to, ataxia, headache, nausea, vomiting, apathy, irritability, fatigue, altered consciousness, high signal intensity on the corpus callosum on fluid attenuation inversion recovery and T2 sequences on cranial MRI, blood-brain barrier disruption, microbleeds in the corpus callosum region on susceptibility-weighted imaging, decreased PaO2, and hypoxemia.

[0085] In one embodiment, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof for the prevention and / or treatment of HAPE. Symptoms of HAPE include, but are not limited to, dry cough, fatigue, weakness, chest tightness and dyspnea, shortness of breath even with mild activity or at rest, cyanosis and coughing up pink frothy sputum, elevated protein levels in bronchoalveolar lavage fluid, a large increase in red blood cells, white blood cells, and neutrophils in bronchoalveolar lavage fluid, recruitment of mononuclear lymphocytes into the airway lumen, scattered alveolar hemorrhages, fibrin exudation in the vascular walls, and hyaline membrane formation.

[0086] In one embodiment, this disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analogue or derivative thereof in the preparation of an anti-altitude hypoxia drug. The anti-altitude hypoxia drug is used to improve physiological indicators in subjects, including but not limited to: blood gas parameters, blood oxygen content, and arterial partial pressure of oxygen.

[0087] In one embodiment, this disclosure provides the use of edaravone and pharmaceutically acceptable salts or analogs or derivatives thereof in the prevention and / or treatment of pulmonary hypertension (PAH) caused by acute high-altitude hypobaric hypoxia. The pulmonary hypertension is involved in the formation of acute high-altitude pulmonary edema (HAPE) and high-altitude pulmonary hypertension (HAPH).

[0088] In one embodiment, this disclosure provides the use of edaravone and its pharmaceutically acceptable salts or analogs or derivatives in the prevention and / or treatment of high-altitude pulmonary hypertension (HAPH). Symptoms of high-altitude pulmonary hypertension include, but are not limited to: right heart catheterization at the altitude of the settlement, mean pulmonary artery pressure >30 mmHg or pulmonary systolic pressure >50 mmHg; right ventricular hypertrophy; heart failure; moderate hypoxemia without erythrocyte polycythemia. Pathological changes in high-altitude pulmonary hypertension include, but are not limited to: 1) vascular endothelial cell damage and proliferation; 2) proliferation and phenotypic transformation of vascular smooth muscle cells; 3) non-muscular arteriolar myogenesis; 4) adventitia fibroblast proliferation and extracellular matrix deposition.

[0089] In one embodiment, this disclosure provides the use of edaravone or its pharmaceutically acceptable salts or analogues or derivatives for reducing inflammatory responses induced by acute high-altitude hypobaric hypoxia. The inflammatory responses include, but are not limited to, changes in the expression of inflammatory factors such as thromboxane A2 (TXA2), endothelin-1 (ET-1), interleukin-1β (IL-1β), hypoxia-inducible factor-1 (HIF-1), tumor necrosis factor (TNF-α), interferon-C, and erythropoietin (EPO), as well as alveolar wall thickening, pulmonary inflammatory cell infiltration, and pulmonary fluid exudation. Detailed Implementation

[0090] The present disclosure 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 disclosure will become clearer and more apparent.

[0091] The term “exemplary” as used herein means “serving as an example, embodiment, or illustration.” Any embodiment illustrated herein as “exemplary” is not necessarily to be construed as superior to or better than other embodiments. Although various aspects of embodiments are shown in the accompanying drawings, the drawings are not necessarily drawn to scale unless specifically indicated otherwise.

[0092] Furthermore, the technical features involved in the different embodiments of this disclosure described below can be combined with each other as long as they do not conflict with each other.

[0093] To further understand this disclosure, preferred embodiments will be described below. These descriptions are merely illustrative of the features and advantages of the technical solutions of this disclosure and are not intended to limit the scope of protection of this disclosure.

[0094] Example

[0095] Partial abbreviations

[0096] MDA: Malondialdehyde.

[0097] SOD: Superoxide dismutase.

[0098] MRI: Magnetic Resonance Imaging.

[0099] HVR: Hypoxic ventilatory response.

[0100] EDA: Edaravone.

[0101] ACZ: Acetazolamide.

[0102] LWC: Lung water content.

[0103] BALF: Bronchoalveolar lavage fluid.

[0104] PaO2: Arterial partial pressure of oxygen.

[0105] pH: Acidity / alkalinity (potential of hydrogen).

[0106] Example 1: Edaravone treatment of high-altitude injury induced by acute high-altitude hypoxia in rats

[0107] Acute altitude sickness, including acute mountain sickness (AMS), is a series of rapidly progressive hypoxia symptoms triggered when a person moves from a low-altitude area to a high-altitude area. Due to the increasing demand for travel or work in high-altitude areas, the need for treatment of acute altitude sickness has increased, requiring effective treatment methods. Currently, there are no reports on the efficacy of edaravone in the treatment or prevention of altitude sickness.

[0108] The applicant discovered through testing that edaravone can improve the survival rate of rats in extremely low-pressure, low-oxygen environments.

[0109] First, the applicant conducted an experiment on the survival of rats in an extreme low-pressure hypoxic environment simulating an altitude of 10,000 meters. After maintaining the pressure simulating an altitude of 10,000 meters for 120 minutes, the pressure was reduced, and the mortality rate was statistically analyzed. In the medicated group (n = 23), edaravone 10 mg / kg, the active ingredient API (Active Pharmaceutical Ingredient) EDA 10 mg / kg + excipient Soluplus 150 mg / kg were administered. In the placebo group (n = 23), Soluplus 150 mg / kg was administered. All medications were given to the rats by gavage 30 minutes before entering the simulation chamber.

[0110] The experimental data showed that 6 rats survived in the group given edaravone, with a survival rate of 26%. Only 3 rats survived in the placebo group, with a survival rate of 13%.

[0111] The results showed that the survival of rats in the medicated group in the extreme low-pressure hypoxic environment for 120 minutes was significantly better than that in the placebo group, thus proving that edaravone has a positive effect on improving the survival rate of animals in an extremely hypoxic environment.

[0112] The purpose of this experiment was to detect the therapeutic effect of edaravone as a preventive and / or therapeutic agent on acute high-altitude injury caused by acute high-altitude hypobaric hypoxia.

[0113] 1.1 Materials and Methods

[0114] 1.1.1 Experimental Animals

[0115] 72 SPF-grade SD rats, half male and half female, with a body weight of 250 - 310 g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the animal production license number: SCXK (Beijing) 2021 - 0011. Before the experiment, the rats were adaptively fed for 5 days. During this period, the mice could freely obtain water and food. The relative humidity of the animal feeding environment was 40% - 70%, the temperature was 20°C - 24°C, and the day and night were each half.

[0116] All animal experiments in this disclosure were approved by the Laboratory Animal Management and Use Committee of Qinghai University, and the ethical batch number was IACUC - DWZX - 2022 - 610.

[0117] 1.1.2 Experimental Instruments and Equipment

[0118] Optical microscope (Nikon Corporation, Japan), experimental animal low-pressure simulation chamber (AVIC Guizhou Fenglei Aviation Ordnance Co., Ltd.), electronic balance (Shanghai Youke Instrument Co., Ltd.), electrothermal constant temperature forced air drying oven (Shanghai Senxin Test Instrument Co., Ltd.), handheld blood gas analyzer (Abbott Laboratories, USA), high-speed low-temperature centrifuge (Hunan Xiangyi Co., Ltd.), multi-functional microplate reader (Tecan Group Ltd., Switzerland). Open field experiment (Shanghai Xinruan Co., Ltd.).

[0119] 1.1.3 Main Reagents and Consumables

[0120] Blood gas and biochemistry multi-test cards (Abbott Laboratories, USA), OCT frozen section embedding medium (Surrey, UK), and the activities of stress markers malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GSH-px), and glutathione S-transferase (GSH-ST) were detected using MDA activity assay kit (No. A003-1-2), SOD activity assay kit (No. A001-3-2), glutathione activity assay kit (No. A006-2-1), GSH activity assay kit (No. A005-1-2), and GSH-ST assay kit (No. A004-1-1), purchased from Jiancheng Company (Nanjing, China).

[0121] 1.2 Experimental Methods

[0122] 1.2.1 Animal grouping and administration

[0123] Seventy-two SD rats, 36 males and 36 females, were randomly divided into four groups according to sex: normal control group (A), hypoxia model group (B), low-dose EDA group (C), medium-dose EDA group (D), high-dose EDA group (E), and acetylcholine group (F). Twelve rats were placed in each group and acclimatized in an SPF rat house for 5 days.

[0124] Table 2

[0125] 1.2.2 Methods for establishing a rat model of injury caused by acute high-altitude hypoxia

[0126] Groups A and B were administered Soluplus via gavage at a standard dose of 100 mg / kg / day. Group C was administered a low-dose EDA solution via gavage at a standard dose of 5 mg / kg / day. Group D was administered a medium-dose EDA solution via gavage at a standard dose of 10 mg / kg / day. Group E was administered a high-dose EDA solution via gavage at a standard dose of 20 mg / kg / day. Group F was administered ACZ solution via gavage at a standard dose of 200 mg / kg / day. Control group rats were housed in the laboratory of the Plateau Medical Center of Qinghai University.

[0127] The medication was administered via gavage twice a day, at 8:00 AM and 8:00 PM on the first and second days, respectively.

[0128] From day 3 to day 5, the experimental animals were placed in a low-pressure simulation chamber (altitude 6000m). To avoid the effects of repeated pressure changes, the twice-daily gavage was combined into a once-daily gavage, administered at 8:00 AM every day for 3 days.

[0129] The edaravone oral solid dispersion particles used were prepared according to Example 7 of CN_105816423_B.

[0130] 1.2.3 Blood gas parameters measurement

[0131] Arterial blood PaO2, PaCO2, SaO2, and pH values ​​were measured using a handheld blood gas analyzer. Before exiting the oxygen chamber, rats in each group were anesthetized by intraperitoneal injection of 2% sodium pentobarbital solution (40 mg / kg) in an oxygen chamber at an altitude of 3500 m. 0.3 mL of blood was collected from the abdominal aorta, and arterial blood gases, including arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), arterial oxygen saturation (SaO2), and pH, were measured using a blood gas analyzer.

[0132] 1.2.4 Hypoxia Indicators in Rat Serum, Brain, and Lung Tissue

[0133] After modeling, rats were anesthetized by intraperitoneal injection of 0.4% sodium pentobarbital. Blood was collected from the abdominal aorta and injected into an anticoagulant tube. The rats were then fixed in a supine position on a surgical board to expose the thoracic cavity. Heart, lung, and brain tissues were quickly harvested, rinsed with 4°C saline, placed in sterile, enzyme-free cryovials, and stored in liquid nitrogen at -80°C for subsequent indicator testing.

[0134] Antioxidant enzymes and other indicators in the serum, brain, and lung tissue of rats in the hypoxic rat experiment were measured according to the kit instructions.

[0135] 1. Blood parameters

[0136] Blood samples were allowed to stand at room temperature for 30 minutes, then centrifuged at 4°C and 3000 rpm for 10 minutes. The supernatant was collected and stored for serum marker detection. MDA, T-SOD, and GSH levels, as well as GSH-Px and GST activities, were measured according to the instructions of each kit.

[0137] 2. Brain tissue indicators

[0138] Brain tissue homogenate was prepared from the cortical portion of rat brain tissue, centrifuged at 3000 r / min for 10 min, and the supernatant was placed in EP tubes. The contents of MDA, T-SOD, and GSH, as well as the activities of GSH-Px and GST, were detected by the TBA method (thiobarbituric acid method), microplate method, and colorimetric method according to the kit instructions.

[0139] 2. Lung tissue parameters

[0140] A portion of rat lung tissue was used to prepare a lung tissue homogenate. The homogenate was centrifuged at 3000 r / min for 10 min, and the supernatant was placed in an EP tube. The contents of MDA, T-SOD, and GSH, as well as the activities of GSH Px and GST, were detected by the TBA method, microplate method, and colorimetric method according to the kit instructions.

[0141] 1.2.5 Lung water content (LWC) measurement

[0142] Take the upper lobe of the right lung of a rat, wash off the bloodstains on the surface in ice-cold PBS solution, blot the moisture of the lung lobe with filter paper, weigh it three times on an electronic balance, and calculate the average value, which is recorded as the wet lung mass (g). Wrap the lung lobe completely in aluminum foil and place it in a 6-well plate. Bake it continuously in a constant temperature drying oven at 50℃ for 72 hours. After the mass is constant, weigh it three times again and calculate the average value, which is recorded as the dry lung mass. Calculate LWC (%) according to formula (1). LWC (%) = wet lung mass / dry lung mass × 100% (1)

[0143] 1.2.6 Lung tissue pathology

[0144] Rat lung tissue was taken, fixed in 4% paraformaldehyde, rinsed, and then processed through dehydration, clearing, and paraffin embedding to prepare paraffin-embedded modules. After being cut into paraffin sections, the lung tissue was stained with hematoxylin and eosin (HE), dehydrated, mounted, and observed under a microscope.

[0145] The Smith scoring method was used to semi-quantitatively analyze pulmonary edema, alveolar and interstitial inflammation, alveolar and interstitial hemorrhage, atelectasis, and hyaline membrane formation, with scores ranging from 0 to 4. No injury was scored as 0 points, lesion area <25% = 1 point, lesion area 25%-50% = 2 points, lesion area 50%-75% = 3 points, and lesion covering the entire field of view = 4 points. The total lung injury score was the sum of the above scores. Ten high-power fields of view were observed for each animal, and the average score was taken.

[0146] 1.2.7 Bronchoalveolar lavage fluid (left lung)

[0147] After the last administration, rats were anesthetized by intraperitoneal injection of 10% chloral hydrate. Their thoracic cavity was cut open and the trachea was immediately exposed. The rat trachea was separated, and a gavage tube was inserted between the third and fourth tracheal rings to a depth of about 2 cm. 2.5 mL of pre-cooled physiological saline was slowly infused, and the bronchoalveolar lavage fluid was repeatedly aspirated 3 times. The protein concentration and bronchoalveolar lavage fluid count of the collected bronchoalveolar lavage fluid were determined using a BCA kit.

[0148] 1.3 Statistical Analysis Methods

[0149] Data were processed using SPSS 22.0 statistical software and plotted using GraphPad Prism 10.1. Quantitative data conforming to a normal distribution were expressed as mean ± standard deviation. This indicates that when the variances of multiple groups are homogeneous, one-way ANOVA is used; when the variances are unequal, the Kruskal-Wallis H test is used; pairwise comparisons are performed using the LSD-t test; and pairwise comparisons between groups with significant differences are performed using the least significant difference (LSD) method. P < 0.05 is considered statistically significant (the specific method is selected according to the data type of the added experimental items).

[0150] result

[0151] 1. EDA's effect on correcting pulmonary function, metabolic state, and acid-base balance caused by acute hypobaric hypoxia.

[0152] Arterial blood gas (ABG) analysis is commonly used in the intensive care unit (ICU), emergency department, and respiratory departments for the diagnosis and assessment of patients' conditions. It can quickly provide information on lung function, metabolic status, and acid-base balance.

[0153] Following prophylactic administration of edaravone, the blood oxygen saturation of hypoxic rats significantly increased. The effect of EDA on the arterial blood oxygen partial pressure (PaO2) of rats exposed to low-pressure hypoxia is shown in Table 3 and Figure 1A, and the pH status of rats in each group is shown in Figure 1B.

[0154] Table 3. Statistical analysis of the effects of different doses of EDA on the partial pressure of oxygen in rat blood (mean ± SE) Note: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001; compared with group E, △△ indicates P < 0.01.

[0155] The results showed that, as shown in Table 3 and Figure 1A, compared with the normal control group A, the PaO2 of rats in group B exposed to hypoxia was significantly reduced (P<0.001); compared with group B, the PaO2 of groups C, D, and E, which were given different doses of EDA, and group F, which was given acetazolamide, was significantly increased (P<0.01).

[0156] In different EDA dosage groups, PaO2 showed a trend of increasing with increasing dosage; in particular, the PaO2 of the medium-dose EDA group and the high-dose EDA group was higher than that of the acetazolamide group, while the PaO2 level of the low-dose group was similar to that of the acetazolamide group.

[0157] Figure 1B shows that there were no significant differences in blood pH among the groups of rats.

[0158] Discussion: Compared with the normal group, the blood PaO2 of rats in the model group exposed to hypobaric hypoxia was significantly reduced, indicating that hypobaric hypoxia can cause a decrease in the partial pressure of oxygen in the blood, PaO2. The experimental results suggest that pre-administration of edaravone for two days, followed by continued administration, can alleviate lung function damage caused by acute hypobaric hypoxia, improve metabolic status, and correct acid-base balance. Doses of 5, 10, and 20 mg / kg / day all achieved the aforementioned effects, with the highest efficacy observed at the high dose of 20 mg / kg / day. Particularly in the high-dose EDA group, the blood PaO2 value of rats was significantly increased (P<0.05), suggesting a protective effect.

[0159] 2. EDA reduces pulmonary edema damage caused by acute hypobaric hypoxia.

[0160] The wet / dry ratio of lung tissue (lung tissue W / D index) indicates the water content of lung tissue. The higher the water content, the greater the degree of edema and the more severe the lung damage. The W / D index can be used to assess the severity of pulmonary edema; the more severe the lung damage, the higher the W / D index.

[0161] The effect of EDA on the W / D index of lung tissue in rats exposed to hypoxia is shown in Table 4 and Figure 2. "All" here means both males and females are included, and the same applies below.

[0162] Table 4. Statistical analysis of the effect of EDA on the wet-to-dry ratio of lung tissue in rats exposed to hypoxia (mean ± SE) Note: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001; compared with group E, △ indicates P < 0.05.

[0163] The results showed that, as indicated in Table 4 and Figure 2, the W / D ratio of group B exposed to hypoxia was significantly increased compared to the normal control group A (P<0.001). Compared to group B, the W / D ratios of groups C, D, and E, which received different doses of EDA, and group F, which received acetazolamide, were significantly reduced. Among these, the improvements were most significant in group D (medium dose EDA), group E (high dose EDA), and group F (acetazolamide) (P<0.001), and the improvement in group E (high dose EDA) was better than that in group F (acetazolamide) (P<0.05).

[0164] Discussion: Compared with the control group, exposure to acute hypobaric hypoxia can cause pulmonary edema, and EDA can reduce hypoxia-induced pulmonary edema. In particular, when EDA was administered at 20 mg / kg / day, the improvement was more significant compared with ACZ treatment at 200 mg / kg / day, suggesting the excellent effect of EDA in reducing pulmonary edema caused by hypobaric hypoxia.

[0165] 3. Effects of EDA on lung tissue in rats induced by acute hypobaric hypoxia

[0166] MDA is produced by lipid peroxidation, and SOD is also an antioxidant enzyme that can scavenge free radicals of superoxide anions. By monitoring the levels of MDA and SOD, it is possible to effectively reflect whether the antioxidant enzyme system of various organs in the body is in a balanced state.

[0167] The effects of EDA on indicators in the lung tissue of rats exposed to hypoxia are shown in Tables 5-6 and Figures 3-4.

[0168] Table 5. Indicators (mean ± SE) in lung tissue of rats exposed to hypoxia. Note: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.01; compared with group B, # indicates P < 0.05, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05, △△△ indicates P < 0.001.

[0169] Table 6. Indicators (mean ± SE) in lung tissue of rats exposed to hypoxia. Note: Compared with group A, * indicates P < 0.05, *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ### indicates P < 0.001; compared with group E, △ indicates P < 0.01, △△ indicates P < 0.01.

[0170] As shown in Tables 5-6 and Figures 3-4, compared with the normal control group A, the GSH content, GSH-PX enzyme activity, and GST activity in the lung tissue of rats exposed to low-pressure hypoxia were significantly reduced (P<0.001), T-SOD activity was significantly reduced (P<0.001), and MDA content was significantly increased (P<0.001).

[0171] GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity

[0172] Compared with group B, groups C, D, and E, which received different doses of EDA, and group F, which received ACZ, all showed significantly higher levels of GSH, GSH-PX enzyme activity, GST activity, and T-SOD activity. Specifically, compared with group B, groups C, D, and E showed significantly increased levels of GSH, GSH-PX enzyme activity, GST activity, and T-SOD activity (P < 0.001).

[0173] When comparing groups C, D, E, and F, group E, which was given 20 mg / kg / d of EDA, showed significantly higher GSH-PX enzyme activity and T-SOD activity compared to group F, which was given 200 mg / kg / d of acetazolamide (P<0.001).

[0174] MDA content

[0175] Compared with model group B, the MDA content in groups C, D, and E, which were given different doses of EDA, and group F, which was given ACZ, was significantly reduced (P<0.001). There was a significant difference in MDA content between groups E and F (△ indicates P<0.01), and the lung tissue of rats given high doses of EDA had even lower MDA content.

[0176] When comparing groups C, D, E, and F, the MDA content in group E, which was given 20 mg / kg / d of EDA, was significantly lower than that in group F, which was given 200 mg / kg / d of acetazolamide (P<0.05).

[0177] Discussion: Compared with the normal group, the lung tissue GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of rats in the model group exposed to hypoxia were significantly decreased, close to half of those in the normal group; while compared with the normal group, the lung tissue MDA content of rats in the model group exposed to hypoxia was increased.

[0178] After treatment with EDA or ACZ, the above indicators all recovered to varying degrees, with significant differences. Furthermore, it was found that compared to ACZ at 200 mg / kg / day, EDA at 20 mg / kg / day (high concentration group) had a stronger effect in increasing GSH-PX enzyme activity, MDA content, and T-SOD activity. In summary, EDA is significantly superior to ACZ in increasing GSH-PX enzyme activity, MDA content, and T-SOD activity. The high dose of EDA at 20 mg / kg / day showed a significant, concentration-dependent effect.

[0179] 4. Effects of EDA on brain tissue hypoxia in rats induced by acute hypobaric hypoxia

[0180] MDA is a metabolite produced by lipid peroxidation of polyunsaturated fatty acids in biological membranes after they are attacked by oxygen free radicals.

[0181] The effects of EDA on brain tissue hypoxia in rats exposed to hypobaric hypoxia are shown in Tables 7-8 and Figures 5-6.

[0182] Table 7. Indicators (mean ± SE) in brain tissue of rats exposed to hypoxia. Note: Compared with group A, *** indicates P < 0.001, ** indicates P < 0.01, and * indicates P < 0.05; compared with group B, ### indicates P < 0.001, ## indicates P < 0.01, and # indicates P < 0.05; compared with group E, △ indicates P < 0.05, and △△△ indicates P < 0.001.

[0183] Table 8. Indicators (mean ± SE) in brain tissue of rats exposed to hypoxia. Note: Compared with group A, * indicates P < 0.05, *** indicates P < 0.001; compared with group B, ## indicates P < 0.01, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05.

[0184] As shown in Tables 7-8 and Figures 5-6, compared with the normal control group A, the brain tissue of rats exposed to hypoxia in group B showed a significant decrease in GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity (P<0.001), and a significant increase in MDA content (P<0.001).

[0185] GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity

[0186] Compared with group B, groups C, D, and E, which received different doses of EDA, and group F, which received ACZ, all showed significantly higher levels of GSH, GSH-PX enzyme activity, GST activity, and T-SOD activity. Specifically, compared with group B, groups C, D, and E showed significantly increased levels of GSH, GSH-PX enzyme activity, GST activity, and T-SOD activity (P < 0.001). Among groups C, D, E, and F, compared with group E, group F showed significantly decreased levels of GSH, GSH-PX enzyme activity (p < 0.001), GST activity, and T-SOD activity (P < 0.05).

[0187] MDA

[0188] Compared with model group B, the MDA content in the brains of groups C, D, and E, which were given different doses of EDA, was significantly reduced (P<0.001). Compared with group E, which was given a high dose of EDA, group F had a higher MDA content, and the difference was statistically significant (P<0.05), indicating an increased degree of lipid peroxidation.

[0189] Discussion: The above results indicate that hypoxia can lead to a decrease in GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity in brain tissue, while increasing MDA content.

[0190] After treatment with EDA and ACZ, all of the above indicators showed significant recovery. At the same time, the high concentration of EDA had a more significant effect on GSH content, GSH-PX enzyme activity, GST activity, MDA content, and T-SOD activity than ACZ. It can be seen that EDA is significantly better than ACZ in terms of GSH content, GSH-PX enzyme activity, GST activity, MDA content, and T-SOD activity.

[0191] In the hypoxic-hypoxic model group, the SOD activity in the brain tissue of rats after 72 hours of hypoxia was lower than that in the ACZ group and the edaravone treatment group, while the SOD activity in the edaravone treatment group was higher than that in the positive drug group. This suggests that edaravone treatment can alleviate brain tissue damage to some extent in rats with brain injury caused by acute hypoxic-hypoxic conditions.

[0192] High-dose EDA reduced hypoxia-induced brain tissue damage in rats, and this effect was stronger than that of ACZ. In the hypoxia model group, after 72 hours of hypoxia, the MDA activity in the brain tissue of rats was higher than that in the ACZ-positive drug group and the edaravone treatment group, while the edaravone treatment group was lower than that in the ACZ-positive drug group. In conclusion, these results suggest that edaravone can be an effective treatment for high-altitude cerebral edema.

[0193] 5. Effects of EDA on serum markers in rats induced by acute hypobaric hypoxia

[0194] The effects of EDA on brain tissue hypoxia in rats exposed to hypobaric hypoxia are shown in Tables 9-10 and Figures 7-8.

[0195] Table 9. Serum markers (mean ± SE) in rats exposed to hypoxia. Note: Compared with group A, *** indicates P<0.001; compared with group B, ### indicates P<0.001, # indicates P<0.05. Table 10: Serum indices (mean±SE) of rats exposed to hypoxia. Note: Compared with group A, *** indicates P < 0.001; compared with group B, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05.

[0196] As shown in Tables 9-10, compared with the normal control group A, the rats in group B exposed to low-pressure hypoxia had significantly reduced serum GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity (P<0.001), and significantly increased MDA content (P<0.001).

[0197] Serum GSH level, GSH-PX enzyme activity, GST activity and T-SOD activity

[0198] Compared with group B, groups C, D, and E, which received different doses of EDA, and group F, which received ACZ, all showed significantly higher serum GSH levels, GSH-PX enzyme activity, GST activity, and T-SOD activity (P < 0.001). Among groups C, D, E, and F, group E, which received 20 mg / kg / d of EDA, showed significantly higher GSH-PX enzyme activity and T-SOD activity compared to group F, which received 200 mg / kg / d of acetazolamide (P < 0.001).

[0199] Serum MDA content

[0200] Compared with group B, groups C, D, E, and F showed a significant reduction in MDA content (P<0.001); among groups C, D, E, and F, group E, which was given 20 mg / kg / d of EDA, had a significantly lower MDA content than group F, which was given 200 mg / kg / d of acetazolamide (P<0.05).

[0201] Discussion: The results above indicate that hypoxia can lead to a decrease in GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity in brain tissue, while increasing MDA content. After prevention and treatment with EDA and ACZ, all of the above indicators showed significant recovery. Furthermore, the high concentration of EDA (20 mg / kg / day) had a more significant effect on GSH-PX enzyme activity, MDA content, and T-SOD activity compared to ACZ. Therefore, EDA is significantly more effective than ACZ in affecting GSH-PX enzyme activity, MDA content, and T-SOD activity.

[0202] summary

[0203] Numerous studies have demonstrated that HH significantly exacerbates the accumulation of excess reactive oxygen species (ROS) and MDA in the brain, reduces GSH levels, and inhibits the activity of T-SOD, GSH, GSH-Px, and GST. This is consistent with the results of the above embodiments. This disclosure finds that edaravone protects cells from damage caused by acute hypobaric hypoxia, suggesting that edaravone can address damage caused by acute altitude sickness.

[0204] 6. Improvement of lung morphology / pulmonary inflammation in rats exposed to hypobaric hypoxia by EDA

[0205] Following prophylactic administration of edaravone, HE staining of the lungs of hypoxic rats showed varying degrees of relief from lung tissue structural damage. The effects of EDA on lung tissue morphology in rats exposed to hypobaric hypoxia are shown in Table 11 and Figure 9. Table 11 shows the Smith scores of lung tissue in each group of rats; "all" in parentheses means both males and females are included. Figure 9 shows HE-stained sections of lung tissue from each group of rats.

[0206] Table 11. Protective effect of EDA on lung tissue injury in rats exposed to hypobaric hypoxia (mean ± SE) Note: Compared with group A, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001; compared with group B, # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001.

[0207] HE staining results

[0208] As shown in Figure 9, compared with the normal control group A, the alveolar walls of rats exposed to low-pressure hypoxia in group B were significantly thickened, the alveolar septa were significantly reduced, and the infiltration of inflammatory cells was significantly increased.

[0209] Compared with model group B, the alveolar wall thickness was significantly reduced, the alveolar interstitial spaces were significantly increased, and the infiltration of inflammatory cells was significantly reduced in the lung tissues of groups C, D, and E, which were given different doses of EDA, and group F, which was given ACZ.

[0210] Meanwhile, comparisons among groups C, D, E, and F showed that group E, which received 20 mg / kg / d of EDA, had the greatest reduction in alveolar walls, the largest alveolar spaces, and the least inflammatory cell infiltration, and was morphologically closest to group A. In groups C, D, and E, which received 5, 10, and 20 mg / kg / d of EDA, the degree of lung tissue lesions decreased sequentially. Group F, which received 200 mg / kg / d of ACZ, was morphologically close to the lesion level of group D, which received 10 mg / kg / d of EDA.

[0211] Furthermore, based on HE-stained sections, the inventors observed the differences between male and female rats in the same group and found that: the pathological changes in the lung tissue of female rats in group A showed a similar trend to those in male rats; however, the degree of lung lesions in female rats in model group B was less than that in male rats in group B; and the male rats in group E, which were given 20 mg / kg / d of EDA, were morphologically similar to the female rats in group E. This suggests that male rats have a weaker tolerance to acute hypobaric hypoxia than female rats, and acute hypobaric hypoxia is more likely to cause hypoxic injury in the same amount of time.

[0212] Smith rating

[0213] As can be seen from the overall Smith score results in Table 11, compared with the normal control group A, the Smith scores of rats in group B exposed to low-pressure hypoxia were significantly higher (P<0.001).

[0214] Compared with model group B, statistically, groups E (administered 20 mg / kg / d EDA) and F (administered 200 mg / kg / d ACZ) showed significantly lower Smith scores, with a notable difference, particularly in group E (P < 0.001). Meanwhile, compared with model group B, groups C and D (administered 5 mg / kg / d EDA, 10 mg / kg / d EDA, respectively) also showed a gradual decrease in damage scores, although no statistically significant difference was observed. This may be because the Smith score is a semi-quantitative analysis, and when scoring tissue sections, it is difficult for the Smith score to differentiate between distinct tissue differences.

[0215] Comparisons among groups C, D, E, and F show no statistically significant differences in Smith scores between any two groups. Numerically, group F's Smith score falls between groups D and E. The reason for this lack of statistical difference is likely due to the Smith score's difficulty in differentiating tissue differences.

[0216] Compared to group E, group F had a slightly higher Smith score, but the difference was not statistically significant.

[0217] Discussion: The experiment shows that exposure to low-pressure hypoxia can induce significant lung tissue damage. Administration of EDA can alleviate lung tissue damage caused by acute hypoxia. The most significant improvement in lung tissue damage was observed when EDA was administered to rats at a dose of 20 mg / kg / day. The effect of EDA at doses of 10-20 mg / kg / day in alleviating lung tissue damage in rats was similar to that of ACZ at 200 mg / kg / day, demonstrating very good preventative and therapeutic effects.

[0218] 7. EDA detection of lung injury in rats induced by acute hypobaric hypoxia (BALF protein concentration)

[0219] Compared with the control group, the bronchoalveolar lavage fluid (BALF) of the high-altitude hypobaric model group showed a large amount of protein exudation. The effects of EDA on the bronchoalveolar lavage fluid (BALF) of rats exposed to hypobaric hypoxia are shown in Table 12 and Figure 10.

[0220] Table 12. Statistical analysis of BALF protein concentration in bronchoalveolar lavage fluid of rats exposed to hypoxia (mean ± SE) Note: Compared with group A, *** indicates P < 0.001; compared with group B, ### indicates P < 0.001; compared with group E, △ indicates P < 0.05.

[0221] As shown in Table 12 and Figure 10, compared with the normal control group A, the BALF protein concentration of rats exposed to low-pressure hypoxia in group B was significantly increased (P<0.001).

[0222] Compared with model group B, the BALF protein concentration was significantly reduced in groups C, D, and E, which were given different doses of EDA, and in group F, which was given ACZ (P<0.001), indicating an effective therapeutic effect.

[0223] The comparison among groups C, D, E, and F showed that group E had a lower BALF protein concentration compared to group F (P<0.05). Numerically, the BALF protein concentration showed a gradual decreasing trend from group C and D to group E, although there was no statistically significant difference.

[0224] Discussion: The results above indicate that acute hypobaric hypoxia can lead to severe pulmonary edema and inflammation in rats, resulting in increased protein concentration in bronchoalveolar lavage fluid (BALF). When EDA and ACZ were administered for prevention and treatment, BALF protein concentration decreased, alleviating pulmonary edema and inflammation. Specifically, treatment with 20 mg / kg / day of EDA significantly reduced BALF protein concentration compared to ACZ at 200 mg / kg / day, demonstrating that EDA is significantly superior to ACZ in improving BALF protein concentration.

[0225] 8. Detection of lung injury in rats induced by acute hypobaric hypoxia using EDA (BALF cell counting)

[0226] Microscopic examination and classification of the obtained bronchoalveolar lavage fluid revealed that all BF groups showed predominant recruitment of mononuclear lymphocytes (MN) cells and secondary recruitment of polymorphonuclear leukocytes (PMN) cells (neutrophils) in the airway lumen.

[0227] The effects of EDA on various cells in the bronchoalveolar lavage fluid of rats exposed to hypoxia, including the number and proportion of white blood cells (WBC), MN#, and PMN#, are shown in Table 13 and Figure 11.

[0228] Table 13. Cell counts in bronchoalveolar lavage fluid of rats exposed to hypoxia (mean ± SE) Note: Compared with group A, *** indicates P < 0.001, * indicates P < 0.05; compared with group B, ### indicates P < 0.001, # indicates P < 0.05; compared with group E, △△ indicates P < 0.01.

[0229] Compared with the normal control group A, the levels of white blood cells (WBC) and neutrophils (PMN) in the bronchoalveolar lavage fluid of the model group B exposed to hypoxia were significantly increased (P<0.001).

[0230] WBC white blood cells, MN

[0231] Compared with model group B, groups C, D, and E, which were given different doses of EDA, and group F, which was given ACZ, showed a significant decrease in WBC and MN (P<0.001).

[0232] Among groups C, D, and E, the white blood cell (WBC) and MN levels in the bronchoalveolar lavage fluid of rats decreased sequentially from group C to groups D and E. Compared with group F, which was given ACZ, group E had significantly lower WBC levels (P<0.01).

[0233] PMN

[0234] Compared with model group B, the PMN values ​​of groups C, D, and E, which were given different doses of EDA, and group F, which was given ACZ, were all reduced to varying degrees, although only group E showed a significant difference in PMN (P<0.001).

[0235] Among groups C, D, and E, the PMN count in the bronchoalveolar lavage fluid of rats decreased sequentially from group C to groups D and E. The PMN count in group F, which was given 200 mg / kg / d of ACZ, was similar to that in the medium-dose EDA group and lower than that in the high-dose EDA group (20 mg / kg / d).

[0236] Discussion: The results showed that prophylactic and therapeutic administration of EDA to rats exposed to hypoxia could reduce the pulmonary inflammatory response caused by acute hypoxia, with higher doses showing the best effect and being superior to the positive control drug ACZ.

[0237] in conclusion

[0238] Experimental results showed that when edaravone was administered to rats in a simulated acute environment at an altitude of 6,000 m, the levels of SOD, GST, and GSH-PX in their serum, lung, and brain tissues significantly increased, while the levels of MDA significantly decreased. Blood gas analysis indicated that the blood oxygen saturation in the edaravone-treated group was significantly higher than that in the model group. HE staining results showed that the level of pulmonary inflammation and fluid exudation in the edaravone-treated group was significantly reduced, indicating that the structural damage to lung tissue was alleviated to varying degrees.

[0239] Experimental results showed that the higher the dose of edaravone, the more significant the improvement effect, suggesting that edaravone has anti-hypoxia activity and a protective effect against major organ damage in rats rapidly ascending to high altitudes. These results suggest that edaravone has the potential to become a novel drug for the prevention and treatment of acute high-altitude hypoxia (HAIs).

[0240] The above results suggest that prophylactic and therapeutic edaravone administration can alleviate acute hypoxemia-associated muscular atrophy (AMS), increase blood oxygen concentration, and reduce hypoxemia. Prophylactic and therapeutic edaravone administration can reduce hepatocellular carcinoma (HACE) and decrease cerebral inflammation. Prophylactic and therapeutic edaravone administration can reduce high-altitude hypobaric hypoxia (HAPE), decrease edema, and reduce lung damage and inflammation. Prophylactic and therapeutic edaravone administration can reduce the inflammatory response induced by acute high-altitude hypobaric hypoxia, decrease pulmonary inflammatory cell infiltration, and reduce inflammatory fluid exudation.

[0241] The present disclosure has been described above with reference to preferred embodiments; however, these embodiments are merely exemplary and illustrative. Various substitutions and modifications can be made to the present disclosure based on these embodiments, all of which fall within the protection scope of the present disclosure.

Claims

1. Use of edaravone or its pharmaceutically acceptable salts or analogs or derivatives in the preparation of medicaments for the prevention and / or treatment of acute high-altitude illnesses.

2. The application according to claim 1, wherein the acute high-altitude disease is at least one selected from the group consisting of: acute mountain sickness (AMS), high-altitude acute brain injury (HACE), and high-altitude acute lung injury (HAPE).

3. The application according to claim 2, wherein the AMS includes mild AMS, moderate to severe AMS, or altitude sickness symptoms expressed as a Lake Louise score of 3-12.

4. The application according to claim 2, wherein the acute high-altitude disease is selected from HACE and HAPE.

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

6. The application according to claim 5, wherein, The administration route of the drug is selected from intravenous infusion, intramuscular injection, oral administration, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or vaginal administration, with oral administration being preferred.

7. The application according to claim 1, wherein, The drug is in the form of a solid dosage form, preferably a solid dispersion.

8. The application according to claim 7, wherein, The solid dispersion formulation includes the active ingredient edaravone or its pharmaceutically acceptable salt or analogue or derivative, a polymer carrier, and optionally a surfactant.

9. The application according to claim 8, 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.

10. The application according to claim 8, wherein, Surfactants include anionic, cationic, or amphoteric surfactants, and are 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.

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

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

13. The application according to claim 1, wherein, The dosage of the edaravone or its pharmaceutically acceptable salts or analogs or derivatives is 0.1-100 mg / kg, preferably 1-24 mg / kg.

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