Application of edaravone in prevention and treatment of acute high-altitude diseases

Abstract

The invention provides application of edaravone and pharmaceutically acceptable salts thereof in prevention and / or treatment of acute high altitude diseases caused by acute plateau hypobaric hypoxia. The acute high altitude diseases include, for example, acute altitude stress (AMS), plateau acute brain injury (HACE), and plateau acute lung injury (HAPE). In addition, the present disclosure provides the use of edaravone and pharmaceutically acceptable salts thereof for the prevention and / or treatment of inflammatory responses caused by plateau pulmonary arterial hypertension (HAPH) and acute plateau hypobaric hypoxia.

Description

Technical Field

[0001] The present disclosure belongs to the pharmaceutical field, and specifically relates to a new use of edaravone, and in particular to the use of edaravone in preparing a drug for preventing and / or treating high-altitude diseases. Background Art

[0002] The harsh environment of low pressure and hypoxia in high and extremely high altitude areas and the increasingly severe climate change have become the main problems restricting human survival and development in this area.

[0003] Hypoxia refers to a pathological process in which the metabolism, function and morphological structure of tissues undergo abnormal changes due to insufficient oxygen supply or oxygen utilization disorders. For patients with long-term or severe hypoxia, it can cause failure of important organs such as the heart and brain, and eventually lead to death.

[0004] At plateaus above 3,500 meters above sea level, people experience high altitude sickness such as chest tightness, shortness of breath, nausea and vomiting, headache and palpitations due to the low pressure and low oxygen environment, which can be life-threatening in severe cases. Therefore, improving one's own hypoxia tolerance is of great significance for people to adapt to plateau life.

[0005] With the growing demand for plateau tourism, sports competitions, urban construction and military activities, more and more people are moving from plains to plateaus. Research on drug prevention and treatment of body damage caused by plateau hypoxia has become a hot topic today.

[0006] Currently, there are few drugs for the treatment of high-altitude illnesses, and acetazolamide is the only drug approved by the FDA for the prevention and treatment of altitude sickness. The Wilderness Medical Society of the United States reported that acetazolamide and dexamethasone can be used to treat symptoms of altitude sickness, and to prevent and treat acute mountain sickness (AMS) and acute high-altitude brain injury (HACE) (Wilderness Medical Society Clinical Practice Guidelines for the Prevention, Diagnosis, and Treatment of Acute Altitude Illness: 2024 Update). However, the effective doses of acetazolamide used in the literature are not consistent, and acetazolamide and dexamethasone have obvious toxic and side effects, which limits their use in routine prevention.

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

[0008] There is a need in this field for anti-acute hypobaric hypoxia drugs with reliable efficacy, which is of great significance to the health and safety of people exposed to hypoxia in plateaus. Summary of the invention

[0009] The inventors found that when animals were rapidly placed under high altitude conditions after being given edaravone for preventive administration, their blood oxygen saturation was significantly increased compared with animals not given the drug for preventive administration, and the damage to the lung tissue structure was alleviated to varying degrees. This method has a protective effect against edema of the brain, lungs and other organs caused by rapid placement under high altitude conditions, and reduces the inflammatory response therein.

[0010] The purpose of the present disclosure is to provide a new use of edaravone, namely, for preventing and / or treating acute high altitude diseases, including AMS, HACE and HAPE.

[0011] First, the present disclosure provides a use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof (preferably as the sole active ingredient) in the preparation of a medicament for preventing and / or treating acute high altitude illness.

[0012] 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). The AMS includes mild AMS, moderate-severe AMS, or mountain sickness symptoms represented by a Lake Louise score of 3-12. In one embodiment, the acute high altitude disease is selected from HACE and HAPE.

[0013] In another aspect, the present disclosure further provides a pharmaceutical composition for preventing and / or treating acute high altitude illness, comprising edaravone or a pharmaceutically acceptable salt or analog or derivative thereof, and one or more pharmaceutically acceptable excipients.

[0014] According to the present disclosure, the pharmaceutical composition is administered by intravenous infusion, intramuscular injection, oral administration, transdermal administration, sublingual administration, intranasal administration, intraocular administration, inner ear administration, rectal administration, or intravaginal administration. Preferably, the pharmaceutical composition is administered orally.

[0015] In one embodiment of the present disclosure, the oral pharmaceutical dosage form is a solid preparation, preferably a solid dispersion preparation, which includes an 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 an optional 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, and 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.

[0016] The present disclosure relates to a method for preventing and / or treating acute high altitude disease using edaravone or a pharmaceutically acceptable salt or analog or derivative thereof, comprising administering a therapeutically effective amount of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof to a subject in need thereof. The acute high altitude disease includes: AMS, HACE and HAPE, etc.

[0017] The present disclosure also relates to edaravone or a pharmaceutically acceptable salt, analog or derivative thereof, and a drug for preventing and / or treating acute high altitude illness, wherein the drug comprises edaravone or a pharmaceutically acceptable salt, analog or derivative thereof.

[0018] The present disclosure unexpectedly discovered that edaravone has a very good preventive and therapeutic effect on acute high-altitude diseases, especially has an outstanding therapeutic effect on high-altitude acute brain injury and high-altitude acute lung injury caused by acute high-altitude hypoxia, and can improve brain damage, blood oxygen concentration, degree of lung tissue damage, degree of pulmonary edema, etc.

[0019] In addition, the present disclosure provides use of edaravone and pharmaceutically acceptable salts thereof for preventing and / or treating inflammatory responses caused by high altitude pulmonary hypertension (HAPH) and acute high altitude hypobaric hypoxia. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly describe the technical solution of the present disclosure, a brief introduction will be given below in conjunction with the accompanying drawings. Obviously, these drawings are only some specific implementation methods recorded in the present disclosure. The present disclosure includes but is not limited to these drawings.

[0021] Figure 1 Results of blood gas analysis of rats exposed to hypobaric hypoxia and given different doses of EDA. Figure 1A is the blood oxygen partial pressure PaO2 of rats exposed to hypobaric hypoxia and given 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 1 B is the pH value of the blood of the rat.

[0022] Figure 2 The dry-to-wet ratio of lung tissue in rats given different doses of EDA and exposed to hypobaric hypoxia. 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 These are the hypoxia indexes of the lung tissue of rats given different doses of EDA and exposed to hypobaric hypoxia. Symbols in the figure: compared with group A, * indicates P < 0.05, **P < 0.01, ***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 These are the hypoxia indexes of the lung tissue of rats given different doses of EDA and exposed to hypobaric hypoxia. Symbols in the figure: compared with group A, * indicates P<0.05, ***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 These are the hypoxia indexes of brain tissue in rats given different doses of EDA and exposed to hypobaric hypoxia. 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 These are the hypoxia indexes of brain tissue in rats given different doses of EDA and exposed to hypobaric hypoxia. 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 The hypoxia index in the serum of rats exposed to hypobaric hypoxia and given different doses of EDA. Symbols in the figure: Compared with group A, *** means P < 0.001; compared with group B, ### means P < 0.001, # means P < 0.05.

[0028] Figure 8 The hypoxia index in the serum of rats exposed to hypobaric hypoxia and given different doses of EDA. Symbols in the figure: compared with group A, *** means P < 0.001; compared with group B, ### means P < 0.001; compared with group E, △ means P < 0.05.

[0029] Fig. 9 HE-stained sections of lung tissues of rats given different doses of EDA and exposed to hypobaric hypoxia.

[0030] Fig.10 The protein concentration in bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia and given different doses of EDA. Symbols in the figure: compared with group A, *** means P < 0.001; compared with group B, ### means P < 0.001; compared with group E, △ means P < 0.05.

[0031] Fig.11 The cell counts of bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia and given 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. DETAILED DESCRIPTION OF THE INVENTION

[0033] definition

[0034] High-altitude sickness: refers to the disease caused by the human body's inability to adapt to the plateau environment when entering a plateau or going from a plateau to a higher altitude. It can be divided into acute high-altitude illnesses (Acute High-altitude illnesses) and chronic high-altitude sickness (Chronic mountain sickness, CMS).

[0035] Acute high altitude illness includes acute mountain sickness (AMS), high altitude acute brain injury (HACE) and high altitude acute lung injury (HAPE). Chronic high altitude illness manifests itself in excessive erythrocytosis and significant pulmonary hypertension, with a prolonged course and eventually right heart failure.

[0036] Acute mountain sickness (AMS) is a variety of pathological reactions caused by the body being exposed to a low oxygen environment in a short period of time after entering a plateau area.

[0037] The diagnosis of AMS is mainly based on symptoms. The symptoms of AMS are atypical and mainly include headache, loss of appetite, nausea, fatigue, dizziness or vertigo, etc. These symptoms usually appear within a few hours to 3 days after arriving at high altitude.

[0038] One of the references for the diagnosis of AMS can be the Lake Louise Acute Mountain Sickness Score (Lake Louise). The LakeLouise score includes the scores of four main symptoms: headache, gastrointestinal symptoms, fatigue or weakness, and dizziness or vertigo, with 0 indicating no discomfort, 1 indicating mild, 2 indicating moderate, and 3 indicating 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]

[0041] AMS can further develop into more severe high-altitude acute brain injury (HACE) and high-altitude acute lung injury (HAPE). It is reported that the destruction of the blood-brain barrier leads to HACE, and the increase in pulmonary capillary pressure and related pressure failure trigger HAPE.

[0042] HACE (high altitude cerebraledema). The diagnosis of HACE is also mainly based on symptoms. Ataxia is usually the earliest clinical manifestation of HACE, which also includes headache, nausea, vomiting, apathy, irritability, fatigue and impaired consciousness. In areas with better medical conditions, other possible diagnoses such as cerebrovascular accident, carbon monoxide poisoning and brain trauma can be ruled out through cranial imaging and laboratory tests.

[0043] When HACE occurs, cranial MRI examination usually shows high signal of the corpus callosum on fluid attenuated inversion recovery and T2 sequences; at the same time, microbleeding foci consistent with blood-brain barrier destruction can also be observed on susceptibility-weighted imaging, mainly in the corpus callosum region.

[0044] HAPE (high altitude pulmonary edema) is a high-permeability edema caused by high flow, high flow rate and high pressure, a life-threatening non-cardiogenic pulmonary edema.

[0045] HAPE patients will experience dry cough, fatigue, weakness, chest tightness and dyspnea. As the disease progresses, patients will feel shortness of breath during slight activities or even rest. They may also experience cyanosis and cough up pink foamy sputum. The amount of protein in the alveolar lavage fluid of patients with high-altitude pulmonary edema is significantly higher than that of the control group and contains a large number of red blood cells, white blood cells and macrophages. Autopsy found that the amount of protein in the pulmonary edema fluid increased significantly; there was scattered bleeding in the alveoli; there was fibrin exudation and hyaline membrane formation in the blood vessel wall. Some reports believe that HAPE is mainly caused by increased pulmonary artery pressure, and inflammatory response is the result, which in turn further aggravates HAPE.

[0046] Acute high-altitude hypobaric hypoxia: in the present disclosure refers to hypobaric hypoxia within 1-7 days or several weeks after arriving at a plateau or high altitude area.

[0047] Inflammatory response caused by acute high-altitude hypobaric hypoxia: In the present disclosure, the inflammatory response includes the inflammatory response involved in pulmonary edema and cerebral edema in HACE and HAPE.

[0048] PAH (pulmonary arterial 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, which leads to increased pulmonary vascular resistance. High pulmonary vascular resistance can lead to a progressive increase in pulmonary artery pressure and sustained vasoconstriction, resulting in poorly adaptable right ventricular hypertrophy. PAH is the basic factor for the occurrence of high altitude pulmonary edema. The pulmonary artery pressure of patients with high altitude pulmonary edema is significantly higher than that of normal people at the same altitude, whether in the early stages of the disease or in the recovery stage. When the altitude increases, the atmospheric pressure decreases, PiO2 decreases accordingly, PaO2 decreases, SaO2% decreases, the body develops hypoxemia, the pulmonary vessels show a hypoxic contraction response, PVR (pulmonary vascular resistance) increases, and PAP (prostatic acid phosphatase) increases with altitude.

[0049] HAPH (high-altitude pulmonary hypertension) is a pathological syndrome of pulmonary vasoconstriction and pulmonary vascular remodeling caused by hypoxia. It is a chronic high-altitude disease with high morbidity and mortality in plateau areas. Related studies have shown that long-term hypoxic stimulation can promote the development of HAPH through vascular remodeling (the main factor) and inflammatory response. The pathological characteristics of HAPH are mainly manifested as: 1) vascular endothelial cell damage and proliferation; 2) proliferation and phenotypic conversion of vascular smooth muscle cells; 3) muscularization of non-muscular arterioles; 4) proliferation of adventitial fibroblasts and deposition of extracellular matrix.

[0050] HH: hypobaric hypoxia. The oxygen concentration under atmospheric pressure in high altitude areas remains basically unchanged, but as the atmospheric pressure decreases, the oxygen partial pressure (PIO2) of the inspired gas decreases proportionally, forming hypobaric hypoxia. In this disclosure, "hypobaric hypoxia" is used synonymously with "hypobaric hypoxia" and "hypobaric hypoxia".

[0051] PaO2: arterial partial pressure of oxygen (PaO2)

[0052] PIO2: partial pressure of oxygen inhaled gas, calculated by PIO2 = FiO2 × (P B -P H2O ). Among them, FiO2 is the inhaled oxygen concentration, P H2O P is the airway water vapor pressure, usually 47 mmHg; B is 1 standard atmosphere.

[0053] Acetazolamide (ACZ) is a carbonic anhydrase (CA) inhibitor that increases the excretion of bicarbonate and promotes diuresis. It also stimulates the respiratory tract, increases arterial oxygen tension, reduces cerebrospinal fluid production, and promotes ion transport across the blood-brain barrier. ACZ is recommended for high-altitude illness by the Wilderness Medical Society Clinical Practice Guidelines for the Prevention, Diagnosis, and Treatment of Acute Altitude Illness: 2024 Update.

[0054] The term "pharmaceutically acceptable" means approved for use in animals, preferably in humans, by a regulatory agency such as EMEA (Europe) and / or FDA (US) and / or any other national regulatory agency.

[0055] The term "excipient" refers to a diluent, adjuvant, or carrier with which a therapeutic agent is administered. Examples of suitable pharmaceutical excipients are described in "Remington's Pharmaceutical Sciences" by EW Martin.

[0056] Soluplus: A polyethylene oxide-polyethylene polymer copolymer, with a molecular formula of (C8H13NO·C4H6O2·C2H4O)x and a chemical formula of (PEO)m(PPO)n(PEO)m, where PEO stands for polyethylene oxide and PPO stands for polypropylene ether. Also known as PCL-PVAc-PE. Soluplus is a solvent-based solubilizer.

[0057] Compound

[0058] 1. Edaravone and its salts, analogs or derivatives

[0059] Edaravone (EDA) is a pyrazolone derivative, its chemical name is 3-methyl-1-phenyl-2-pyrazol-5-one, molecular formula C 10 H 10 N2O, the structural formula is as follows:

[0060]

[0061] The core structure or active center of EDA is 2-pyrazol-5-one, which plays a major role in free radical scavenging activity.

[0062] EDA is a nitrogen-containing heterocyclic compound, which is alkaline and easily reacts with acid to form salts. Its pharmaceutically acceptable salts include inorganic acid salts and organic acid salts commonly found in the art, wherein inorganic acid salts include hydrochlorides, carbonates, phosphates, etc., and organic acid salts include citrates, tartrates, acetates, oxalates, salicylates, malates, lactates, etc.

[0063] EDA analogs include methyl at the 3-position of the pyrazoline ring, which may be ethyl, propyl or other lower (C 1-6) alkyl, or lower alkoxy, such as methoxy, ethoxy, etc.; or the methyl at position 3 can be H, and the H at position 4 can be substituted by a lower alkyl or alkoxy. The derivatives of edaravone include esters, that is, the ketone at position 5 of the pyrazoline ring is converted into an enol, which reacts with a carboxylic acid to form an ester, such as a methyl ester, an ethyl ester, etc. The ester (precursor) is hydrolyzed in vivo and then converted into a ketone. In addition, the phenyl group is optionally substituted by one or more substituents selected from lower alkyl, lower alkoxy, nitro, halogen, etc.

[0064] EDA derivatives include Tables 1-2 and Figure 2 As many as 18 EDA derivatives are disclosed in 1 ), position 3 (R 3 ) and position 4 (R 4 ) can be further modified to have similar oxidation potential (Epa) and hydroxyl radical scavenging activity (IC 50 ).

[0065] 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).

[0066] EDA derivatives also include compounds of formula (I) disclosed in Chinese Patent Application No. 201710036907.7, especially compounds of formula (Ia) (BE). The contents of this patent application document are fully incorporated into this disclosure by reference.

[0067] 2. Dosage forms of edaravone

[0068] Edaravone and its salts or analogs or derivatives can be developed into various dosage forms according to the needs of the administration route, for example, they can be administered by intravenous infusion, intramuscular injection, oral, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or vaginal routes.

[0069] Edaravone was first developed by Mitsubishi Tanabe Pharm Corp. (Osaka, Japan) to improve neurological symptoms, daily living activities and functional impairment caused by acute cerebral infarction. The marketed dosage form is injection. At present, Edaravone has been widely used in the treatment of acute ischemic stroke (AIS) and amyotrophic lateral sclerosis (ALS) at home and abroad, and is also used to treat diseases related to excessive reactive oxygen species (ROS), such as cardiovascular disease.

[0070] CN105816423B discloses various preparations of edaravone (e.g., solid phase dispersion preparations) and their use in effectively treating human oxidative stress-related diseases; CN 110996944A discloses that a liquid aqueous solution of edaravone is used to treat 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 that edaravone or its analogs or derivatives are used to treat cerebral amyloid angiopathy (CAA) (the contents of the above three patent application documents are all incorporated into this disclosure by reference).

[0071] Taking into account the convenience of administration and increasing the patient's medication compliance, the preferred dosage form is a solid preparation, preferably a solid dispersion preparation, which is administered orally.

[0072] The solid dispersion formulation may include an active ingredient, edaravone or a pharmaceutically acceptable salt thereof or an analog or derivative thereof, a polymer carrier and an optional 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.

[0073] The surfactant includes an anionic, anionic or an amphoteric surfactant 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.

[0074] The preparation method of the oral preparation disclosed in the present invention can be prepared by referring to conventional methods in the art, and it is particularly preferred to prepare the oral preparation of edaravone or a pharmaceutically acceptable salt thereof by referring to the method disclosed in CN105816423B.

[0075] In the present disclosure, edaravone or its pharmaceutically acceptable salts or analogs or derivatives are sometimes collectively referred to as edaravone and used interchangeably.

[0076] According to the present disclosure, the dosage of edaravone is between 0.1 mg / kg and 100 mg / kg. For treatment, the dosage 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.

[0077] When used to prevent high-altitude diseases, it is recommended that the subject use the disclosed preparation 2 days before or 1 day before starting mountaineering activities at an altitude of less than 1,200 meters, and continue to use it. The optional dosage is 1-2 times a day. When used to treat high-altitude diseases, the subject starts using the disclosed preparation when the altitude increases rapidly and / or symptoms of AMS, HACE, and HAPE appear, and continues to use it until the symptoms of the disease are partially or completely improved. The optional dosage is once every 12 hours, or once every 24 hours, and the usage is adjusted according to the symptoms.

[0078] The use of the preparation disclosed herein may be optionally combined with the following measures: stopping the subject from ascending to an altitude, lowering the altitude, giving the subject oxygen, using a portable hyperbaric oxygen chamber, etc.

[0079] The dosage and frequency of administration of the disclosed formulations can vary depending on the patient's condition, and may sometimes require relatively high doses (e.g., 5.0-35 mg / kg) at relatively short intervals until the progression of the disease is alleviated or stopped, preferably until the patient shows partial or complete improvement in disease symptoms.

[0080] The disclosed drugs can be administered parenterally, topically, intravenously, orally, subcutaneously, intraarterially, intracranially, intrathecally, intraperitoneally, intranasally or intramuscularly. 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 arm or leg muscles.

[0081] use

[0082] The present disclosure provides use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof in the preparation of a medicament for preventing and / or treating acute high altitude illness.

[0083] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof in the preparation of a medicament for preventing and / or treating acute high altitude diseases caused by acute high altitude hypobaric hypoxia, wherein the acute high altitude diseases include acute mountain sickness (AMS), high altitude acute brain injury (HACE) and high altitude acute lung injury (HAPE), etc.

[0084] In one embodiment, the AMS disclosed herein includes mild AMS, moderate-severe AMS, or altitude sickness symptoms represented by a Lake Louise score of 3 to 12. Those skilled in the art will appreciate that the AMS disclosed herein does not include further aggravated symptoms.

[0085] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof for preventing and / or treating AMS, and / or reducing the Lake Louise score of a subject. The symptoms of AMS include but are not limited to headache, loss of appetite, nausea, fatigue, dizziness or vertigo, decreased PiO2, and hypoxemia.

[0086] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof for preventing and / or treating HACE. The symptoms of HACE include, but are not limited to, ataxia, headache, nausea, vomiting, apathy, irritability, fatigue, impaired consciousness, high signal of the corpus callosum on fluid-attenuated inversion recovery and T2 sequences on cranial MRI, blood-brain barrier disruption, microbleeding foci in the corpus callosum region on susceptibility-weighted imaging, decreased PaO2, and hypoxemia.

[0087] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof for preventing and / or treating HAPE. The symptoms of HAPE include, but are not limited to, dry cough, fatigue, weakness, chest tightness and dyspnea, shortness of breath during slight activity or even at rest, cyanosis and coughing up pink foamy sputum, increased protein content in bronchoalveolar lavage fluid, a large increase in red blood cells, a large increase in white blood cells, a large increase in neutrophils in bronchoalveolar lavage fluid, recruitment of mononuclear lymphocytes in the airway lumen, scattered alveolar hemorrhage, fibrin exudation and hyaline membrane formation in the vascular wall, etc.

[0088] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof in the preparation of an anti-altitude hypoxia drug. The anti-altitude hypoxia drug is used to improve physiological indicators of a subject including but not limited to: blood gas index, blood oxygen content, arterial oxygen partial pressure, etc.

[0089] In one embodiment, the present disclosure provides the use of edaravone and its pharmaceutically acceptable salts or analogs or derivatives for preventing and / or treating 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).

[0090] In one embodiment, the present disclosure provides the use of edaravone and its pharmaceutically acceptable salts or analogs or derivatives for preventing and / or treating high altitude pulmonary hypertension (HAPH). Symptoms of high altitude pulmonary hypertension include, but are not limited to: right heart catheterization at the altitude of settlement, mean pulmonary artery pressure>30mmHg or pulmonary artery systolic pressure>50mmHg; right ventricular hypertrophy; heart failure; moderate hypoxemia without polycythemia. Pathological changes of high altitude pulmonary hypertension include, but are not limited to: 1) vascular endothelial cell damage and proliferation; 2) proliferation and phenotypic conversion of vascular smooth muscle cells; 3) muscularization of non-muscular arterioles; 4) proliferation of adventitial fibroblasts and deposition of extracellular matrix.

[0091] In one embodiment, the present disclosure provides the use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof for reducing the inflammatory response caused by acute high altitude hypobaric hypoxia. The inflammatory response includes, but is 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, erythropoietin (EPO), alveolar wall thickening, lung inflammatory cell infiltration, lung fluid exudation, etc. DETAILED DESCRIPTION

[0092] The present disclosure is further described in detail below through the accompanying drawings and embodiments. Through these descriptions, the characteristics and advantages of the present disclosure will become more clear and distinct.

[0093] The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments. Although various aspects of the embodiments are shown in the drawings, the drawings are not necessarily drawn to scale unless otherwise noted.

[0094] In addition, the technical features involved in different embodiments of the present disclosure described below can be combined with each other as long as they do not conflict with each other.

[0095] In order to further understand the present disclosure, the preferred embodiments of the present disclosure are described below in conjunction with the embodiments. These descriptions are only examples of the features and advantages of the technical solutions of the present disclosure, and are not intended to limit the protection scope of the present disclosure.

[0096] Example

[0097] Some abbreviations

[0098] MDA: Malondialdehyde.

[0099] SOD: superoxide dismutase.

[0100] MRI: Magnetic Resonance Imaging.

[0101] HVR: hypoxic ventilatory response.

[0102] EDA: Edaravone.

[0103] ACZ: acetazolamide.

[0104] LWC: lung water content.

[0105] BALF: bronchoalveolar lavage fluid.

[0106] PaO2: arterial partial pressure of oxygen.

[0107] pH: acidity (potential of hydrogen).

[0108] Example 1 Edaravone treats high altitude injury caused by acute high altitude hypobaric hypoxia in rats

[0109] Acute high altitude illness includes acute mountain sickness (AMS), which is a series of rapidly progressive hypoxic symptoms caused by people moving from low altitude areas to high altitude areas. Due to the increasing demand for people to travel or work in high altitude areas, the demand for the treatment of acute high altitude illness has increased, and effective treatment methods need to be provided. There is currently no report on the effect of edaravone in the treatment or prevention of high altitude illness.

[0110] The applicant discovered through testing that edaravone can improve the survival rate of rats in an extreme low-pressure and low-oxygen environment.

[0111] First, the applicant conducted an experiment on the survival of rats in an extreme hypobaric hypoxic environment simulating an altitude of 10,000 meters. The pressure at an altitude of 10,000 meters was simulated for 120 minutes, and then the blood pressure was lowered and the mortality rate was counted. The medication group (n=23) was given 10 mg / kg of Edaravone, active ingredient API (Active Pharmaceutical Ingredient) EDA 10 mg / kg + excipient Soluplus 150 mg / kg. The placebo group (n=23) was given Soluplus 150 mg / kg. All drugs were given to rats by gavage 30 minutes before entering the simulation cabin.

[0112] Experimental data showed that 6 rats given edaravone survived, with a survival rate of 26%, while only 3 rats given a placebo survived, with a survival rate of 13%.

[0113] The results showed that the survival rate of rats in the drug group in the extreme low-pressure and low-oxygen environment for 120 minutes was significantly better than that in the placebo group, which proves that edaravone has a positive effect on the survival rate of animals in extreme low-oxygen environments.

[0114] The purpose of this study was to examine the therapeutic effects of edaravone as a preventive and / or therapeutic agent for acute high altitude injury caused by acute high altitude hypobaric hypoxia.

[0115] 1.1 Materials and methods

[0116] 1.1.1 Experimental animals

[0117] 72 SPF SD rats, half male and half female, weighing 250-310 g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., with animal production certificate number: SCXK (Beijing) 2021-0011. The mice were adaptively fed for 5 days before the experiment, during which time they had free access to water and food. The relative humidity of the animal breeding environment was 40%-70%, the temperature was 20℃-24℃, and the day and night were half and half.

[0118] All animal experiments disclosed in this paper were approved by the Experimental Animal Care and Use Committee of Qinghai University, with ethics approval number IACUC-DWZX-2022-610.

[0119] 1.1.2 Experimental instruments and equipment

[0120] Optical microscope (Nikon, Japan), experimental animal hypobaric simulation chamber (AVIC Guizhou Fenglei Aviation Ordnance Co., Ltd.), electronic balance (Shanghai Youke Instrument Co., Ltd.), electric constant temperature blast drying oven (Shanghai Senxin Testing Instrument Co., Ltd.), handheld blood gas analyzer (Abbott, USA), high-speed low-temperature centrifuge (Hunan Xiangyi Company), multifunctional microplate reader (Tecan, Switzerland). Open field experiment (Shanghai Xinruan Co., Ltd.).

[0121] 1.1.3 Main reagents and consumables

[0122] Blood gas and biochemistry multiple test cards (Abbott, USA), OCT frozen section embedding medium (Surrey, UK), 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 detection kit (NO.A004-1-1) were used to detect the activities of stress markers malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GSH-px) and glutathione S-transferase (GSH-ST) and were purchased from Jiancheng Company (Nanjing, China).

[0123] 1.2 Experimental methods

[0124] 1.2.1 Animal grouping and drug administration

[0125] 72 SD rats, including 36 males and 36 females, were randomly divided into normal control group (A), hypoxia model group (B), EDA low-dose group (C), EDA medium-dose group (D), EDA high-dose group (E) and acetazamide group (F) according to gender. There were 12 rats in each group, and the rats in each group were placed in SPF rat room for 5 days for adaptive feeding.

[0126] Table 2

[0127]

[0128] 1.2.2 Methods for establishing a rat model of acute high altitude hypobaric hypoxia injury

[0129] Group A and Group B were given Soluplus by gavage, with a standard dose of 100 mg / kg / d, Group C was given EDA low-dose solution by gavage, with a standard dose of 5 mg / kg / d, Group D was given EDA medium-dose solution by gavage, with a standard dose of 10 mg / kg / d, Group E was given EDA high-dose solution by gavage, with a standard dose of 20 mg / kg / d, and Group F was given ACZ solution by gavage, with a standard dose of 200 mg / kg / d. The rats in the control group were housed in the laboratory of Qinghai University Plateau Medical Center.

[0130] The drug was administered by intragastric administration twice on the first and second days, at 8 am and 8 pm respectively.

[0131] On the 3rd to 5th day, the experimental animals were placed in a low pressure simulation chamber (altitude 6000m). To avoid the effects of repeated increases and decreases in pressure, the two daily gavages were combined into one daily gavage, which was administered at 8 am every day for 3 days.

[0132] The edaravone oral solid dispersion particles used were prepared according to Example 7 of CN_105816423_B.

[0133] 1.2.3 Blood gas index determination

[0134] A handheld blood gas analyzer was used to detect arterial blood PaO2, PaCO2, SaO2, pH and other values. Before leaving the oxygen chamber, each group of rats was anesthetized by intraperitoneal injection of 2% sodium pentobarbital solution (40 mg / kg) in the oxygen chamber at an altitude of 3500 m, and 0.3 mL of abdominal aortic blood was collected. Arterial blood gas was measured using a blood gas analyzer, including arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), arterial oxygen saturation (SaO2), and pH (potential of hydrogen).

[0135] 1.2.4 Hypoxia indexes in rat serum, brain, and lung tissues

[0136] After the modeling was completed, 0.4% sodium pentobarbital was injected intraperitoneally for anesthesia, and blood was collected from the abdominal aorta and injected into an anticoagulant tube. The rats were then fixed in a supine position on the surgical operating board to expose the chest cavity. The heart, lung, and brain tissues were quickly removed, rinsed with 4°C saline, placed in sterile enzyme-free cryopreservation tubes, and stored in liquid nitrogen at -80°C for subsequent index detection.

[0137] The antioxidant enzymes and other indicators in the serum, brain, and lung tissues of rats in the hypoxic rat experiment were measured according to the kit instructions.

[0138] 1. Blood indicators

[0139] The blood was placed at room temperature for 30 minutes, centrifuged at 4°C, 3000 rpm for 10 minutes, and the supernatant was collected and retained for the detection of serum indicators. MDA, T-SOD, GSH content and GSH-Px, GST activity were detected according to the instructions of each kit.

[0140] 2. Brain tissue indicators

[0141] The cortical part of rat brain tissue was taken to prepare brain tissue homogenate, which was centrifuged at 3000r / min for 10min. The supernatant was placed in an EP tube and tested for MDA, T-SOD, GSH content and GSH-Px, GST activity using TBA method (thiobarbituric acid method), microplate method, and colorimetric method according to the instructions of the kit.

[0142] 2. Lung tissue indicators

[0143] Lung tissue of rats was taken to prepare lung tissue homogenate, which was centrifuged at 3000 r / min for 10 min. The supernatant was placed in an EP tube and tested for MDA, T-SOD, GSH content and GSH Px, GST activity using TBA method, microplate method and colorimetric method according to the instructions of the kit.

[0144] 1.2.5 Determination of lung water content (LWC)

[0145] The upper lobe of the right lung of the rat was taken, and the blood on the surface was washed in an iced PBS solution. The water of the lung lobe was absorbed by filter paper, and the weight was weighed 3 times on an electronic balance to calculate the average value, which was recorded as the wet weight of the lung (g). The lung lobe was completely wrapped with tin foil and placed in a 6-well plate, and baked in a 50℃ constant temperature drying oven for 72 hours. After the weight was constant, it was weighed again 3 times to obtain the average value, which was recorded as the dry weight of the lung. It was calculated according to formula (1).

[0146] LWC (%) = wet lung mass / dry lung mass × 100% (1)

[0147] 1.2.6 Lung tissue pathomorphology

[0148] The rat lung tissue was fixed in 4% paraformaldehyde, and after washing, it was dehydrated, transparentized, and wax-impregnated to make a paraffin-embedded module. After cutting into paraffin sections, the lung tissue was stained with HE, dehydrated, sealed, and observed under a microscope.

[0149] The Smith scoring method was used to perform semi-quantitative analysis of pulmonary edema, alveolar and interstitial inflammation, alveolar and interstitial hemorrhage, atelectasis and hyaline membrane formation, with a score of 0-4; no damage was 0 point, lesion range <25% = 1 point, lesion range 25%-50% = 2 points, lesion range 50%-75% = 3 points, lesion full field of view = 4 points. The total lung injury score was the sum of the above items. Ten high-power microscopic fields were observed for each animal and the average value was taken.

[0150] 1.2.7 Bronchoalveolar lavage fluid (left lung)

[0151] After the last administration, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, the chest 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 saline was slowly infused, and the blood was repeatedly drawn and washed three times. The protein concentration and alveolar lavage fluid count of the collected alveolar lavage fluid were determined using a BCA kit.

[0152] 1.3 Statistical analysis methods

[0153] SPSS22.0 statistical software was used to process the data and GraphPad Prism 10.1 was used to draw the data. Quantitative data that conformed to normal distribution were expressed as mean ± standard deviation (X ± SD). One-way analysis of variance was used when the variances were equal among multiple groups. Kruskal-Wallis H test was used when the variances were unequal. LSD-t test was used for pairwise comparison. The least significant difference (LSD) method was used for pairwise comparison between groups when there was a significant difference. P < 0.05 was considered statistically significant (the specific method was selected according to the data type of the added experimental project).

[0154] result

[0155] 1. EDA corrects lung function, metabolic state and acid-base balance caused by acute hypobaric hypoxia

[0156] Arterial blood gas analysis (ABG) is commonly used in the diagnosis and assessment of diseases in the intensive care unit (ICU), emergency department and respiratory department, and can quickly provide information about lung function, metabolic status and acid-base balance.

[0157] After the preventive administration of edaravone, the blood oxygen saturation of the hypoxic rats increased significantly. The effect of EDA on the arterial oxygen partial pressure (PaO2) of rats exposed to hypobaric hypoxia is shown in Table 3. Figure 1 A, pH conditions of rats in each group are shown in Figure 1 B.

[0158] Table 3 Statistics of the effects of different doses of EDA on blood oxygen partial pressure in rats (mean ± SE)

[0159]

[0160] Note: 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

[0161] The results show that from Table 3 and Figure 1 A shows that compared with the normal control group A, the PaO2 of rats in group B exposed to hypobaric hypoxia was significantly reduced (P<0.001); compared with group B, the PaO2 of groups C, D, and E given different doses of EDA and group F given acetazolamide was significantly increased (P<0.01).

[0162] In the groups receiving different doses of EDA, PaO2 showed a trend of increasing with increasing dose; in particular, the PaO2 in the medium-dose EDA group and the high-dose EDA group was higher than that in the acetazolamide group, and the PaO2 levels in the low-dose group and the acetazolamide group were similar.

[0163] Figure 1 B shows that there is no significant difference in blood pH among the rats in each group.

[0164] Discussion: Compared with the normal group, the blood PaO2 of rats in the model group exposed to hypobaric hypoxia was significantly reduced, indicating that the body's blood oxygen partial pressure PaO2 can be reduced in a low-pressure hypoxic environment. The experimental results suggest that pre-administration of edaravone for two days and 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 / d can all achieve the above effects, and the high dose of 20 mg / kg / d has the highest therapeutic effect. Especially in the high-dose EDA group, the blood PaO2 value of rats increased significantly (P<0.05), suggesting a protective effect.

[0165] 2. EDA reduces lung tissue edema damage caused by acute hypobaric hypoxia

[0166] The dry / wet specific gravity of lung tissue (W / D index of lung tissue) can indicate the water content of lung tissue. The higher the water content, the higher the degree of edema and the more serious the damage to the lungs. The W / D index can be used to assess the severity of pulmonary edema. The more serious the lung damage, the higher the W / D index.

[0167] The effect of EDA on the W / D index of rat lung tissue exposed to hypobaric hypoxia is shown in Table 4. Figure 2 The “all” here means that both males and females are included, and the same applies below.

[0168] Table 4 Statistics of the effect of EDA on the dry-wet ratio of lung tissue in rats exposed to hypobaric hypoxia (mean ± SE)

[0169]

[0170] Note: 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. The results show that from Table 4 and Figure 2 It can be seen that compared with the normal control group A, the W / D index of group B exposed to hypobaric hypoxia increased significantly (P<0.001); compared with group B, the W / D of groups C, D, E given different doses of EDA and group F given acetazolamide decreased significantly, among which the improvements in group D with medium dose of EDA, group E with high dose and group F with acetazolamide were the most obvious (P<0.001), and the improvement of group E with high dose of EDA was better than that of group E with acetazolamide (P<0.05).

[0171] Discussion: Compared with the normal group, exposure to acute hypobaric hypoxia can cause pulmonary edema damage, and administration of EDA can reduce hypoxia-induced pulmonary edema damage. In particular, when 20 mg / kg / d EDA was administered, the degree of treatment improvement was more significant compared with 200 mg / kg / d ACZ treatment, suggesting that EDA has an excellent effect in reducing pulmonary edema caused by hypobaric hypoxia.

[0172] 3. Effects of EDA on lung tissue of rats induced by acute hypobaric hypoxia

[0173] MDA is produced by lipid peroxidation. SOD is also an antioxidant enzyme that can scavenge superoxide anion free radicals. By monitoring the levels of MDA and SOD, it can effectively reflect whether the antioxidant enzyme system of each organ in the body is in a balanced state.

[0174] The effects of EDA on the indicators in the lung tissue of rats exposed to hypobaric hypoxia are shown in Tables 5-6. Figure 3-Figure 4 .

[0175] Table 5 Indexes in lung tissue of rats exposed to hypobaric hypoxia (mean ± SE)

[0176]

[0177]

[0178] Note: Compared with group A, * indicates P < 0.05, **P < 0.01, ***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

[0179] Table 6 Indexes in lung tissue of rats exposed to hypobaric hypoxia (mean ± SE)

[0180]

[0181] 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

[0182] From Table 5-Table 6, Figure 3-Figure 4 It can be seen that compared with the normal control group A, the GSH content, GSH-PX enzyme activity, and GST activity in the lung tissue of rats in group B exposed to low-pressure hypoxia were significantly reduced (P<0.001), the T-SOD activity was significantly reduced (P<0.001), and the MDA content was significantly increased (P<0.001).

[0183] GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity

[0184] Compared with the model group B, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of groups C, D and E given different doses of EDA and group F given ACZ were higher, and the differences were statistically significant. In particular, compared with group B, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of groups C, D and E were significantly increased (P<0.001).

[0185] When comparing groups C, D, E and F, the GSH-PX enzyme activity and T-SOD activity of group E given 20 mg / kg / d of EDA were higher than those of group F given 200 mg / kg / d of acetazolamide, and the difference was significant (P<0.001).

[0186] MDA content

[0187] Compared with the model group B, the MDA content in groups C, D and E given different doses of EDA, and group F 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 MDA content in the lung tissue of rats given high doses of EDA was lower.

[0188] 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).

[0189] Discussion: Compared with the normal group, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of the lung tissue of the rats in the model group exposed to hypobaric hypoxia were significantly decreased, close to half of that in the normal group; compared with the normal group, the MDA content of the lung tissue of the rats in the model group exposed to hypobaric hypoxia was increased.

[0190] After EDA or ACZ treatment, the above indicators were restored to varying degrees, with significant differences. At the same time, it can be seen that compared with 200 mg / kg / d of ACZ, 20 mg / kg / d of EDA (high concentration group) had a stronger effect on increasing GSH-PX enzyme activity, MDA content and T-SOD activity indicators. In summary, EDA is significantly better than ACZ in terms of its ability to increase GSH-PX enzyme activity, MDA content and T-SOD activity indicators. The high dose of 20 mg / kg / d of EDA has a significant effect and is concentration-dependent.

[0191] 4. Effect of EDA on brain tissue hypoxia in rats induced by acute hypobaric hypoxia

[0192] MDA is a metabolite produced by lipid peroxidation after polyunsaturated fatty acids in biological membranes are attacked by oxygen free radicals.

[0193] The effects of EDA on brain tissue hypoxia in rats exposed to hypobaric hypoxia are shown in Tables 7 and 8. Figure 5-Figure 6 .

[0194] Table 7 Indexes in brain tissue of rats exposed to hypobaric hypoxia (mean ± SE)

[0195]

[0196] 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, △△△ indicates P<0.001

[0197] Table 8 Indexes in brain tissue of rats exposed to hypobaric hypoxia (mean ± SE)

[0198]

[0199] 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

[0200] From Table 7-Table 8, Figure 5-Figure 6It can be seen that compared with the normal control group A, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of the brain tissue of the rats in group B exposed to hypobaric hypoxia were significantly reduced (P<0.001), and the MDA content was significantly increased (P<0.001).

[0201] GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity

[0202] Compared with group B, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of groups C, D and E given different doses of EDA and group F given ACZ were higher, and the differences were statistically significant. In particular, compared with group B, the GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity of groups C, D and E increased significantly (P<0.001). Comparison between groups C, D, E and F showed that compared with group E, group F had a significant decrease in GSH content, GSH-PX enzyme activity (p<0.001), GST activity and T-SOD activity (P<0.05).

[0203] MDA

[0204] 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, the MDA content in group F was higher, and the difference was statistically significant (P<0.05), indicating an increase in the degree of lipid peroxidation.

[0205] Discussion: The above results show that hypoxia can lead to a decrease in GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity in brain tissue, and an increase in MDA content.

[0206] After treatment with EDA and ACZ, the above indicators were significantly restored. At the same time, the effect of high concentration of EDA on GSH content, GSH-PX enzyme activity, GST activity, MDA content and T-SOD activity was more obvious than that of ACZ. It can be seen that EDA has a significantly better effect on GSH content, GSH-PX enzyme activity, GST activity, MDA content and T-SOD activity than ACZ.

[0207] The SOD activity of brain tissue in the rats in the hypobaric hypoxia model group was lower than that in the ACZ group and the edaravone treatment group at 72 hours of hypoxia, while that in the edaravone treatment group was higher than that in the positive drug group, indicating that the application of edaravone in the treatment of rats with brain damage caused by acute hypobaric hypoxia can alleviate brain tissue damage to a certain extent.

[0208] High-dose EDA reduced the damage of hypoxia to rat brain tissue, and this ability was stronger than ACZ. The MDA activity of brain tissue in the hypoxia model group was higher than that in the ACZ positive drug group and the edaravone treatment group at 72 hours after hypoxia, and the MDA activity in the edaravone treatment group was lower than that in the ACZ positive drug group. In summary, the results suggest that edaravone can be used as an effective drug for the treatment of high-altitude cerebral edema.

[0209] 5. Effects of EDA on serum parameters of rats induced by acute hypobaric hypoxia

[0210] The effects of EDA on brain tissue hypoxia in rats exposed to hypobaric hypoxia are shown in Tables 9 and 10. Figure 7-Figure 8 .

[0211] Table 9 Indicators in serum of rats exposed to hypobaric hypoxia (mean ± SE)

[0212]

[0213] Note: Compared with group A, *** indicates P < 0.001; compared with group B, ### indicates P < 0.001, # indicates P < 0.05 Table 10 Serum indexes of rats exposed to hypobaric hypoxia (mean ± SE)

[0214]

[0215] 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

[0216] As shown in Tables 9 and 10, compared with the normal control group A, the rats in group B exposed to hypobaric hypoxia had a significant decrease in serum 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).

[0217] Serum GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity

[0218] Compared with model group B, groups C, D, E, and ACZ given different doses of EDA and group F had higher serum GSH content, GSH-PX enzyme activity, GST activity, and T-SOD activity, with statistically significant differences (P<0.001). Comparison between groups C, D, E, and F showed that compared with group F given 200 mg / kg / d of acetazolamide, group E given 20 mg / kg / d of EDA had significantly higher GSH-PX enzyme activity and T-SOD activity (P<0.001).

[0219] Serum MDA content

[0220] Compared with group B, the MDA content in groups C, D, E and F was significantly reduced (P<0.001). Comparison among groups C, D, E and F showed that the MDA content in group E, which was given 20 mg / kg / d of EDA, was lower than that in group F, which was given 200 mg / kg / d of acetazolamide, and the difference was significant (P<0.05).

[0221] Discussion: The above results show that hypoxia can lead to a decrease in brain tissue GSH content, GSH-PX enzyme activity, GST activity and T-SOD activity, and an increase in MDA content. After EDA and ACZ prevention and treatment, the above indicators were significantly restored. At the same time, the high concentration of EDA at 20 mg / kg / d had a more obvious effect on GSH-PX enzyme activity, MDA content and T-SOD activity than ACZ. It can be seen that EDA has a significantly better effect on GSH-PX enzyme activity, MDA content and T-SOD activity than ACZ.

[0222] summary

[0223] A large number of studies have shown that HH significantly increases the accumulation of excessive 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 examples. The present disclosure found that edaravone protects cells from damage caused by acute hypobaric hypoxia, suggesting that edaravone can cope with damage caused by acute altitude sickness.

[0224] 6. EDA improves lung tissue morphology / pulmonary inflammation in rats exposed to hypobaric hypoxia

[0225] After the preventive administration of Edaravone, HE staining of the lungs of hypoxic rats showed that the damage to the lung tissue structure was alleviated to varying degrees. The effects of EDA on the morphology of the lung tissue of rats exposed to hypobaric hypoxia are shown in Table 11. Fig. 9 Table 11 shows the Smith scores of lung tissues of rats in each group. The "all" in brackets means that both males and females are included. Fig. 9 HE-stained sections of lung tissues of rats in each group.

[0226] Table 11 Protective effect of EDA on lung tissue damage in rats exposed to hypobaric hypoxia (mean ± SE)

[0227]

[0228]

[0229] Note: 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

[0230] HE staining results

[0231] Depend on Fig. 9 The results showed that compared with the normal control group A, the alveolar walls in the lung tissue of rats in group B exposed to low-pressure hypoxia were significantly thickened, the alveolar septa were significantly reduced, and the infiltration of inflammatory cells was significantly increased.

[0232] Compared with the model group B, the alveolar wall thickness was significantly reduced, the alveolar space was significantly increased, and the inflammatory cell infiltration was significantly reduced in the lung tissues of groups C, D, and E given different doses of EDA and group F given ACZ.

[0233] At the same time, comparison between groups C, D, E and F showed that group E, which was given 20 mg / kg / d of EDA, had the greatest reduction in alveolar wall, the largest alveolar space, the least inflammatory cell infiltration, and was morphologically closest to group A; the degree of lung tissue lesions in groups C, D and E, which were given 5, 10, and 20 mg / kg / d of EDA, decreased successively, and group F, which was given 200 mg / kg / d of ACZ, was morphologically close to the lesion level of group D, which was given 10 mg / kg / d of EDA.

[0234] In addition, according to the 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 had a similar trend to those of male rats; but the degree of lesions in the lung tissue of female rats in group B was smaller than that of 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 damage at the same time.

[0235] Smith score

[0236] From the overall Smith score results in Table 11, it can be seen that compared with the normal control group A, the Smith scores of the rats in group B exposed to hypobaric hypoxia were increased, and the difference was significant (P<0.001).

[0237] Compared with model group B, the Smith scores of group E, which was given 20 mg / kg / d EDA, and group F, which was given 200 mg / kg / d ACZ, were statistically lower, especially group E, which was significantly reduced (P<0.001). At the same time, compared with model group B, groups C and D, which were given 5 and 10 mg / kg / d EDA, also showed a gradual decrease in injury scores, although no statistical difference was obtained. This may be because the Smith score is a semi-quantitative analysis, and when scoring the slices, it is difficult to widen the score interval in distinguishing tissue differences.

[0238] It can be seen that there is no statistical difference in Smith score between groups C, D, E and F. From the numerical point of view, the Smith score of group F is between group D and group E. It is speculated that the reason for not obtaining statistical difference is the same as above, which may be due to the difficulty of Smith score in distinguishing tissue differences.

[0239] Compared with group E, the Smith score in group F was slightly higher, but there was no significant difference.

[0240] Discussion: The experiment shows that exposure to hypobaric hypoxia can cause obvious lesions in lung tissue. Administration of EDA can reduce tissue damage to lung tissue caused by acute hypobaric hypoxia. When EDA is administered to rats at a dose of 20 mg / kg / d, the effect on lung tissue damage is most obvious. When EDA is administered to rats at a dose of 10-20 mg / kg / d, the effect of reducing lung tissue damage is close to that of 200 mg / kg / d of ACZ, and it has a very good preventive and therapeutic effect.

[0241] 7. EDA detection of acute hypobaric hypoxia-induced lung injury in rats (BALF protein concentration)

[0242] Compared with the control group, a large amount of protein exudates were found in the bronchoalveolar lavage fluid (BALF) of the high altitude hypobaric hypoxia model group. The effects of EDA on the bronchoalveolar lavage fluid (BALF) of rats exposed to hypobaric hypoxia are shown in Table 12. Fig.10 .

[0243] Table 12 Statistics of BALF protein concentration in bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia (mean ± SE)

[0244]

[0245] 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

[0246] From Table 12, Fig.10 It can be seen that compared with the normal control group A, the BALF protein concentration of rats in group B exposed to hypobaric hypoxia increased significantly (P<0.001).

[0247] Compared with model group B, groups C, D and E given different doses of EDA, and group F given ACZ showed a significant decrease in BALF protein concentration (P<0.001), indicating an effective therapeutic effect.

[0248] Comparison among groups C, D, E, and F showed that the BALF protein concentration of rats in group E was lower than that in group F (P<0.05). Numerically, the BALF protein concentration showed a trend of gradually decreasing from group C, D to group E, although there was no statistically significant difference.

[0249] Discussion: The above results show that acute hypobaric hypoxia can lead to severe pulmonary edema and inflammatory process in rats, which increases the protein concentration in bronchoalveolar lavage fluid. After EDA and ACZ were given for prevention and treatment, the protein concentration of BALF decreased, and the edema and inflammation of the lungs were alleviated. In particular, after treatment with 20 mg / kg / d of EDA, the effect of EDA in reducing the protein concentration of BALF was more obvious than that of 200 mg / kg / d of ACZ, indicating that EDA is significantly better than ACZ in improving the protein concentration of BALF.

[0250] 8. EDA detection of acute hypobaric hypoxia-induced lung injury in rats (BALF cell count)

[0251] Microscopic examination and classification of the bronchoalveolar lavage fluid revealed that in all BF groups, mononuclear lymphocytes (MN) were mainly recruited in the airway lumen, supplemented by polymorphonuclear leukocytes (PMN) cells (neutrophils).

[0252] The effects of EDA on the cells in the bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia: the number and proportion of white blood cells (WBC), MN#, and PMN# are shown in Table 13. Fig.11 .

[0253] Table 13 Statistics of cells in bronchoalveolar lavage fluid of rats exposed to hypobaric hypoxia (mean ± SE)

[0254]

[0255] 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

[0256] Compared with the normal control group A, the levels of WBC and PMN in the bronchoalveolar lavage fluid of the model group B exposed to hypobaric hypoxia were significantly increased (P<0.001).

[0257] WBC, MN

[0258] Compared with model group B, the WBC and MN in groups C, D and E given different doses of EDA, and group F given ACZ were significantly decreased (P<0.001).

[0259] The WBC and MN values ​​in the bronchoalveolar lavage fluid of rats decreased in order from group C to group D and group E. Compared with group E and group F given ACZ, the WBC value in group E was lower, with a significant difference (P<0.01).

[0260] PMN

[0261] Compared with model group B, the PMN values ​​of groups C, D and E given different doses of EDA, and group F given ACZ were reduced to varying degrees, although only PMN of group E showed significant differences (P<0.001).

[0262] The PMN counts in the bronchoalveolar lavage fluid of rats decreased from group C to group D and group E. The PMN counts in group F, which was given 200 mg / kg / d of ACZ, were similar to those in the medium-dose EDA group, but lower than those in the high-dose EDA group of 20 mg / kg / d.

[0263] Discussion: The results showed that the preventive and therapeutic administration of EDA to rats exposed to hypobaric hypoxia could reduce the pulmonary inflammatory response caused by acute hypobaric hypoxia, and the high dose had the best effect, which was better than the positive drug ACZ.

[0264] in conclusion

[0265] The experimental results showed that when edaravone was used on rats in an acute environment simulating an altitude of 6,000m, the levels of SOD, GST, GSH-PX and other indicators in their serum, lung and brain tissues increased significantly, while the levels of MDA and other indicators decreased significantly. The results of blood gas analysis showed 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 lung inflammation and fluid exudation in the edaravone-treated group was significantly reduced, indicating that the damage to the lung tissue structure was alleviated to varying degrees.

[0266] The experimental results showed that the greater the dose of edaravone, the more obvious the improvement effect, suggesting that edaravone has anti-altitude hypoxia activity and has a protective effect on the main organ damage of rats that are rapidly exposed to the plateau. The results suggest that edaravone has the potential to become a new drug for the prevention and treatment of acute HAIs.

[0267] The above results suggest that preventive and therapeutic administration of edaravone can alleviate AMS, increase blood oxygen concentration, and alleviate hypoxemia. Preventive and therapeutic administration of edaravone can alleviate HACE and reduce brain inflammation. Preventive and therapeutic administration of edaravone can alleviate HAPE, reduce the degree of edema, and alleviate lung damage and inflammation. Preventive and therapeutic administration of edaravone can alleviate the inflammatory response caused by acute high-altitude hypobaric hypoxia, reduce lung inflammatory cell infiltration, and reduce inflammatory fluid exudation.

[0268] The present disclosure has been described above in conjunction with preferred embodiments, but these embodiments are only exemplary and serve only as an illustration. On this basis, various replacements and improvements may be made to the present disclosure, all of which fall within the scope of protection of the present disclosure.

Claims

1. Use of edaravone or a pharmaceutically acceptable salt or analog or derivative thereof in the preparation of a medicament for preventing and / or treating acute high altitude illness.

2. The use 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 use according to claim 2, wherein the AMS comprises mild AMS, moderate to severe AMS, or altitude sickness symptoms represented by a Lake Louise score of 3-12.

4. The use according to claim 2, wherein the acute high altitude illness is selected from HACE and HAPE.

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

6. The use according to claim 5, wherein: The administration method of the drug is selected from intravenous drip, intramuscular injection, oral, transdermal, sublingual, intranasal, intraocular, inner ear, rectal, or intravaginal administration, preferably oral administration.

7. The use according to claim 1, wherein: The dosage form of the drug is a solid preparation, preferably a solid dispersion preparation.

8. The use according to claim 7, wherein: The solid dispersion preparation comprises an active ingredient, edaravone or a pharmaceutically acceptable salt or analog or derivative thereof, a polymer carrier and an optional surfactant.

9. The use 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 use according to claim 8, 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.

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

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

13. The use according to claim 1, wherein: The dosage of edaravone or its pharmaceutically acceptable salt, analog or derivative is 0.1-100 mg / kg, preferably 1-24 mg / kg.

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