Use of masitinib for the treatment of COVID-19

Abstract

The present invention relates to masitinib or a pharmaceutically acceptable salt or solvate thereof for use in treating coronavirus infection, such as SARS-CoV-2 infection that causes COVID-19, in a subject in need thereof.

Description

Technical Field

[0001] The present invention relates to treating coronavirus infection in a subject in need thereof. In particular, the present invention relates to treating SARS-CoV-2 infection in a subject in need thereof, i.e., treating COVID-19, COVID-19-related pneumonia and / or COVID-19-related acute respiratory distress syndrome (ARDS) in a subject in need thereof. Background Art

[0002] Coronaviruses (CoV) are positive-sense single-stranded RNA viruses (+ssRNA viruses) of the Coronaviridae family, characterized by an unusually large RNA genome, a unique replication strategy, and a unique morphology seen under an electron microscope, i.e., a crown-like appearance resulting from rod-like projections protruding from the surface of its envelope (Fehr & Perlman, Methods Mol Biol. 2015; 1282: 1–23). Coronaviruses are nidoviruses (i.e., they belong to the order Nidovirales), infect mammals and birds, and cause a wide range of respiratory, gastrointestinal, nervous system, and systemic diseases.

[0003] Human coronaviruses were first identified in the mid-1960s and were initially thought to cause only mild respiratory infections, such as the common cold, in most cases. Therefore, it is estimated that four endemic human CoVs (HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1) account for 10% to 30% of upper respiratory tract infections in adults (Paules et al., JAMA. 2020 Feb 25; 323 (8): 707-708). However, in recent years, two highly pathogenic coronaviruses that cause severe respiratory diseases have appeared in animal hosts: severe acute respiratory syndrome coronavirus (SARS-CoV), first identified in 2003, and Middle East respiratory syndrome coronavirus (MERS-CoV), first identified in 2012. 8096 cases of severe acute respiratory syndrome (SARS), including 774 deaths, have been reported worldwide, and 1728 cases of Middle East respiratory syndrome (MERS), including 624 deaths, have been reported worldwide (de Wit et al., Nat Rev Microbiol. 2016; 14(8):523–534).

[0004] In December 2019, a new infectious respiratory disease of unknown cause was identified (Huang et al., Lancet. 2020; 395(10223):497–506; Wang et al., Lancet. 2020; 395(10223):470–473; Zhu et al., N Engl J Med. 2020; 382(8):727–733). Coronavirus RNA was rapidly identified in some patients, and in January 2020, researchers from the Shanghai Public Health Clinical Center and the School of Public Health and their collaborators published the full genome sequence of the newly identified human coronavirus SARS-CoV-2 (previously named 2019-nCoV). The genome sequence of SARS-COV-2 has 89% nucleotide identity with the genome sequence of the bat coronavirus SARS-like-CoVZXC21 (SARS-like-CoVZXC21) and 82% nucleotide identity with the genome sequence of human SARS-CoV (Chanet al., Lancet. 2020; 395(10223): 514–523). As previously shown for SARS-CoV, SARS-CoV2 appears to utilize ACE2 (angiotensin-converting enzyme 2) as a receptor for viral cell entry (Hoffmann et al., Cell. 2020Apr16; 181(2): 271-280).

[0005] SARS-CoV2 infection is thought to be asymptomatic or cause few or no clinical manifestations in 30% to 60% of infected subjects. In symptomatic infected subjects, the disease caused by SARS-COV-2 is now referred to as "COVID-19" (coronavirus disease 2019). COVID-19 is a respiratory disease that typically presents first with symptoms including headache, muscle pain, and / or fatigue / tiredness, followed by fever and respiratory symptoms (such as dry cough, shortness of breath, and / or chest tightness). Although symptoms remain mild in most subjects, in others they may progress to pneumonia (referred to herein as COVID-19-associated pneumonia or COVID-19 pneumonia) and / or multi-organ failure. Complications of COVID-19 include acute respiratory distress syndrome (ARDS) (referred to herein as COVID-19-associated ARDS or COVID-19ARDS), RNAaemia, acute cardiac injury, and secondary infection (Huang et al., Lancet. 2020; 395(10223): 497–506). It is estimated that approximately 5% of patients with COVID-19 require hospitalization, with approximately 25% requiring admission to an intensive care unit (ICU). COVID-19 causes substantial morbidity and mortality and is likely to place unprecedented strain on many health systems.

[0006] As a result, global efforts to evaluate new antiviral drugs and therapeutic strategies for the treatment of COVID-19 have intensified. Notably, many clinical trials have been registered to evaluate the efficacy of drugs such as remdesivir (a nucleotide analog antiviral drug under development), lopinavir / ritonavir (an antiretroviral therapy mainly used to treat human immunodeficiency virus 1 (HIV-1)), and chloroquine or hydroxychloroquine (both mainly used to prevent and treat malaria, and also used to treat rheumatoid arthritis and lupus erythematosus). However, there is still a lack of therapeutic drugs with proven efficacy in preventing and / or treating COVID-19, COVID-19-related pneumonia, or COVID-19-related acute respiratory distress syndrome (ARDS).

[0007] Therefore, there remains a need for effective treatments for infections with nested viruses, including coronaviruses, and for infections with picornaviruses, which are +ssRNA viruses that belong to the same class as the nested viruses (i.e., Pisoniviricetes). In particular, there remains a need for effective treatments for infections with coronaviruses, particularly betacoronaviruses. Currently, there is an urgent need for effective and safe treatments for infections with SARS-CoV-2, which causes COVID-19, particularly for prophylactic and / or therapeutic treatments for COVID-19-associated pneumonia and COVID-19-associated acute respiratory distress syndrome (ARDS).

[0008] The present invention relates to a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally in combination with isoquercetin, for treating a nested virus or a small RNA virus infection in a subject in need thereof, in particular for treating a coronavirus infection, such as SARS-CoV-2 infection causing COVID-19. Summary of the invention

[0009] The present invention relates to a 2-aminoaryl thiazole derivative or a pharmaceutically acceptable salt or solvate thereof for use in treating coronavirus infection in a subject in need thereof.

[0010] In one embodiment, the coronavirus infection is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection that causes COVID-19.

[0011] In one embodiment, the 2-aminoarylthiazole derivative or a pharmaceutically acceptable salt or solvate thereof is used for combined administration with isoquercetin, preferably for combined administration with isoquercetin in a dosage range of about 0.4 g / day to about 2 g / day, more preferably for combined administration with isoquercetin in a dosage range of about 1 g / day.

[0012] In one embodiment, the 2-aminoarylthiazole derivative has the formula (II):

[0013]

[0014] in:

[0015] -R 1 are independently selected from hydrogen, halogen, (C 1 -C 10 ) alkyl, (C 3 -C 10) cycloalkyl, trifluoromethyl, alkoxy, amino, alkylamino, dialkylamino, solubilizing groups and solubilizing groups substituted (C 1 -C 10 )alkyl; and

[0016] -m is 0-5.

[0017] In one embodiment, the 2-aminoarylthiazole derivative or a pharmaceutically acceptable salt or solvate thereof is masitinib or a pharmaceutically acceptable salt or solvate thereof. In one embodiment, the pharmaceutically acceptable salt of masitinib is masitinib mesilate.

[0018] In one embodiment, 2-aminoaryl thiazole derivatives or pharmaceutically acceptable salts or solvates thereof are used for oral administration. In one embodiment, 2-aminoaryl thiazole derivatives or pharmaceutically acceptable salts or solvates thereof are administered in a dosage range of about 1 mg / kg / day to about 12 mg / kg / day (mg / kg body weight / day), preferably in a dosage range of about 3 mg / kg / day to about 6 mg / kg / day. In one embodiment, 2-aminoaryl thiazole derivatives or pharmaceutically acceptable salts or solvates thereof are used for administration at an initial dose of about 3 mg / kg / day during at least one week, and thereafter at a dose of about 4.5 mg / kg / day, with toxicity control for each dose increase.

[0019] In one embodiment, the subject exhibits at least one risk factor that may lead to an increased risk of developing COVID-19.

[0020] In one embodiment, the subject has mild to moderate COVID-19, preferably moderate COVID-19. In one embodiment, the subject has severe COVID-19. In one embodiment, the subject has critical COVID-19.

[0021] In one embodiment, the subject has COVID-19 and has a score range of 2 to 9 on the World Health Organization (WHO) COVID-19 10-point progression scale (as described in Table 1 herein). In one embodiment, the subject has COVID-19 and has a score of 2 or 3 on the WHO COVID-19 10-point progression scale (as described in Table 1 herein). In one embodiment, the subject has COVID-19 and has a score range of 4 to 6, preferably 4 or 5, on the WHO COVID-19 10-point progression scale (as described in Table 1 herein). In one embodiment, the subject has COVID-19 and has a score range of 2 to 6, preferably a range of 2 to 5, more preferably 4 or 5 on the modified WHO COVID-19 7-point progression scale (as described in Table 2 herein).

[0022] In one embodiment, the 2-aminoarylthiazole derivative or a pharmaceutically acceptable salt or solvate thereof is used for administration with at least one additional pharmaceutically active agent. In one embodiment, at least one additional pharmaceutically active agent is selected from antiviral agents, anti-interleukin 6 (anti-IL6) agents, protease inhibitors, Janus-associated kinase (JAK) inhibitors, BXT-25, brilacidin, dehydroandrographolide succinate, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon, carrimycin, and any mixture thereof.

[0023] definition

[0024] In the present invention, the following terms have the following meanings:

[0025] "About" preceding a number includes plus or minus 10% or less of the numerical value. It should be understood that the value referred to by the term "about" itself is also specifically and preferably disclosed.

[0026] As used herein, "baseline" refers to the time before the start of treatment with a 2-aminoarylthiazole derivative as described herein, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof. For example, for a given subject, the interleukin 6 (IL6) plasma level at baseline is the interleukin 6 (IL6) plasma level before the subject is administered a 2-aminoarylthiazole derivative as described herein, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof.

[0027] "Best supportive care" refers to the supportive care routinely provided to subjects who are hospitalized and have respiratory diseases, particularly lower respiratory tract diseases, such as pneumonia or ARDS. Best supportive care may include, for example, at least one of the following: supplemental oxygen (O2) (also called oxygen therapy) (e.g., by mask or nasal prong), non-invasive ventilation (NIV), invasive mechanical ventilation, extracorporeal membrane oxygenation (ECMO), vasopressor therapy (e.g., phenylephrine, norepinephrine, epinephrine, vasopressin and / or dopamine), fluid therapy, antimicrobial therapy, renal support, sedation.

[0028] As used herein, "consisting essentially of" with respect to a composition, a pharmaceutical composition or a medicament means that the 2-aminoarylthiazole derivative as described herein, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is the only active agent (also referred to as active ingredient or active compound), i.e., the only agent that exhibits biological or pharmacological activity in the composition, pharmaceutical composition or medicament.

[0029] As used herein, "high flow" or "high flow oxygen" refers to high flow oxygen therapy (HFOT) as a form of respiratory support.

[0030] As used herein, "laboratory confirmed SARS-CoV-2 infection" refers to SARS-CoV-2 infection confirmed by a laboratory test, such as an rRT-PCR (real-time reverse transcription polymerase chain reaction) test that allows detection of the presence of SARS-CoV-2 in a subject sample (e.g., a nasal swab sample, an oropharyngeal swab sample, a sputum sample, a lower respiratory tract aspirate, a bronchoalveolar lavage fluid, a nasopharyngeal wash / aspirate, or a nasal aspirate), or an antibody test (e.g., an enzyme-linked immunosorbent assay (ELISA)) that allows detection of the presence of antibodies against SARS-CoV-2 in a subject sample (e.g., a blood sample).

[0031] A "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to an excipient or carrier that does not produce adverse, allergic or other untoward reactions when administered to a mammal, preferably a human. It includes any and all solvents, such as dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delaying agents. A pharmaceutically acceptable excipient or carrier refers to any type of non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation aid. For human administration, the formulation should meet the sterility, pyrogenicity, general safety and purity standards required by regulatory agencies such as the FDA (U.S. Food and Drug Administration) or EMA (European Medicines Agency).

[0032] "Subject" refers to a mammal, preferably a human. Mammals include, but are not limited to, rodents, including mice; Lagomorpha, including rabbits; Carnivora, including felines (cats) and canines (dogs); Artiodactyla, including bovines (cows) and suines (pigs); Perissodactyla, including equines (horses); Primates, including monkeys, apes and humans. In one embodiment, the mammal is selected from rodents, lagomorpha, carnivora, artiodactyla, perissodactyla and primates. In one embodiment, the mammal is selected from mice, rabbits, cats, dogs, cattle, pigs, horses, monkeys, apes and humans. In one embodiment, the subject is a primate, preferably a human. According to one embodiment, the subject is a mammal, preferably a human, who has been exposed to, suspected of being exposed to, or expected to be exposed to a nested virus or a small RNA virus, in particular a coronavirus such as SARS-CoV-2. According to one embodiment, the subject is a mammal, preferably a human, suffering from a nested virus infection or a picornavirus infection, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19. In one embodiment, the subject may be a "patient", i.e. a mammal, in particular a warm-blooded mammal, preferably a human, who is awaiting or receiving medical care or was / is / will be the subject of a medical procedure, or is being monitored for the development of a nested virus infection or a picornavirus infection, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19.

[0033] "Therapeutically effective amount" or "therapeutically effective dose" refers to the amount or dosage or concentration of a 2-aminoarylthiazole derivative as described herein, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally in combination with isoquercetin or quercetin, sufficient to induce a meaningful benefit in the subject, cell or tissue to be treated. Meaningful benefits include, for example, detectable treatment, alleviation or reduction of one or more symptoms of a disease caused by a nested virus or a small RNA virus (e.g., inflammation, effusion), in particular a coronavirus; inhibition, prevention of development, prevention or cessation of further development of a viral infection or disease caused by a nested virus or a small RNA virus, in particular a coronavirus; reduction of the incidence of a disease caused by a nested virus or a small RNA virus, in particular a coronavirus; prevention of a disease caused by a nested virus or a small RNA virus, in particular a coronavirus, in a subject, cell or tissue at risk of the disease but not yet diagnosed with the disease; and / or detectable inhibition of one or more active sites of a viral protein in a subject, cell or tissue. The meaningful benefit observed in the subject, cell or tissue to be treated can be achieved to any suitable extent (10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more). In one embodiment, the therapeutically effective dose is the amount or dose or concentration of the 2-aminoarylthiazole derivative described herein, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally in combination with isoquercetin or quercetin, which is intended to prevent, reduce, alleviate or slow down (mitigate) one or more symptoms or manifestations of a nested virus infection or a small RNA virus infection, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19, in the subject in need of treatment without causing significant negative or adverse side effects.

[0034] "Treatment" refers to therapeutic treatment, prophylactic (or preventive) treatment, or therapeutic treatment and prophylactic (or preventive) treatment, wherein the purpose is to prevent, reduce, alleviate and / or slow down (mitigate) one or more symptoms or manifestations of a nested virus infection or a small RNA virus infection, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19 in a subject in need thereof. Symptoms of coronavirus infection, in particular SARS-CoV-2 infection that causes COVID-19, include but are not limited to fever and respiratory symptoms, such as dry cough and / or dyspnea that may require respiratory support (e.g., supplemental oxygen, non-invasive ventilation, invasive mechanical ventilation, extracorporeal membrane oxygenation (ECMO)). The manifestations of coronavirus infection, in particular SARS-CoV-2 infection, include but are not limited to the viral load (also referred to as viral load or viral titer) detected in a subject's sample. In one embodiment, "treatment" refers to therapeutic treatment. In another embodiment, "treatment" refers to prophylactic or preventive treatment. In yet another embodiment, "treatment" refers to both prophylactic (or preventive) treatment and therapeutic treatment. In one embodiment, the purpose of treatment according to the present application is to achieve at least one of the following:

[0035] oReduction of viral load detected in subject samples;

[0036] o Reduced need for respiratory support, such as reduced use of ECMO, invasive mechanical ventilation, noninvasive ventilation, or supplemental oxygen (including high-flow oxygen therapy); and / or reduced need for vasopressor therapy;

[0037] o Transfer from the intensive care unit;

[0038] oDischarge from hospital. DETAILED DESCRIPTION OF THE INVENTION

[0040] The present invention relates to 2-aminoarylthiazole derivatives as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a nested virus infection in a subject in need thereof. Examples of nested viruses (i.e., viruses belonging to the order Nested Viruses) include coronaviruses, toroviruses, arteriviruses, and okaviruses. Diseases caused by nested viruses include, but are not limited to, COVID-19, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), respiratory diseases (e.g., pneumonia, bronchitis, pleural effusion), inflammatory diseases (e.g., inflammation, COVID-19-induced inflammation, pediatric multisystem inflammatory syndrome (PMIS)), porcine reproductive and respiratory syndrome, equine viral arteritis, and gastroenteritis).

[0041] In one embodiment, the nested virus is a coronavirus, an arterivirus, or a tegument virus. In one embodiment, the nested virus is a coronavirus.

[0042] Therefore, according to one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a coronavirus infection in a subject in need thereof.

[0043] In one embodiment, the coronavirus is an alpha coronavirus or a beta coronavirus, preferably a beta coronavirus, including a beta coronavirus, a beta coronavirus, a beta coronavirus, a beta coronavirus, and a beta coronavirus. Therefore, in one embodiment, the coronavirus is a beta coronavirus. In one embodiment, the beta coronavirus is a beta coronavirus, a beta coronavirus, a beta coronavirus, or a beta coronavirus.

[0044] Examples of alpha coronaviruses include, but are not limited to, human coronavirus 229E (HCoV-229E) and human coronavirus NL63 (HCoV-NL63) (sometimes also referred to as HCoV-NH or New Haven human coronavirus). Examples of beta coronaviruses include, but are not limited to, human coronavirus OC43 (HCoV-OC43), human coronavirus HKU1 (HCoV-HKU1), Middle East respiratory syndrome-related coronavirus (MERS-CoV) (previously referred to as novel coronavirus 2012 or HCoV-EMC), severe acute respiratory syndrome coronavirus (SARS-CoV) (also referred to as SARS-CoV-1 or classic SARS (SARS-classic)), and severe acute respiratory syndrome coronavirus (SARS-CoV-2). In one embodiment, beta coronavirus is HCoV-OC43, MERS-CoV, SARS-CoV (also referred to as SARS-CoV-1) or SARS-CoV-2. In one embodiment, beta coronavirus is HCoV-sOC43 or SARS-CoV-2.

[0045] In one embodiment, the coronavirus is selected from the group comprising or consisting of HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, MERS-CoV, SARS-CoV-1 and SARS-CoV-2.

[0046] In one embodiment, the coronavirus is selected from the group comprising or consisting of: MERS-CoV, SARS-CoV-1 and SARS-CoV-2. Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein or a pharmaceutically acceptable salt or solvate thereof for use in treating a subject in need thereof with a MERS-CoV coronavirus infection causing MERS, a SARS-CoV-1 infection causing SARS, or a SARS-CoV-2 infection causing COVID-19.

[0047] In one embodiment, the coronavirus is a MERS coronavirus. In one embodiment, the coronavirus is MERS-CoV that causes Middle East Respiratory Syndrome (MERS).

[0048] In one embodiment, the coronavirus is a SARS coronavirus.

[0049] In one embodiment, the coronavirus is SARS-CoV-1 or SARS-CoV-2. Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a SARS-CoV-1 infection causing SARS or a SARS-CoV-2 infection causing COVID-19 in a subject in need thereof.

[0050] In one embodiment, the coronavirus is SARS-CoV (also known as SARS-CoV-1), which causes severe acute respiratory syndrome (SARS).

[0051] According to one embodiment, the coronavirus is SARS-CoV-2, which causes COVID-19. Therefore, according to one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a SARS-CoV-2 infection that causes COVID-19 in a subject in need thereof. In one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating COVID-19 in a subject in need thereof.

[0052] The present invention also relates to 2-aminoarylthiazole derivatives as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a picornavirus infection in a subject in need thereof. Examples of picornaviruses include poliovirus, rhinovirus, enterovirus and coxsackievirus. Diseases caused by picornaviruses include, but are not limited to, acute flaccid myelitis (AFM), respiratory diseases and gastrointestinal diseases.

[0053] In one embodiment, the picornavirus is a poliovirus, a rhinovirus, an enterovirus, or a coxsackievirus. In one embodiment, the picornavirus is a rhinovirus or a coxsackievirus.

[0054] Therefore, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a nested virus infection or a picornavirus infection in a subject in need thereof. In other words, therefore, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in treating an infection caused by a virus of the class Nidoviridae, wherein the virus of the class Nidoviridae is a nested virus or a picornavirus.

[0055] In one embodiment, the severity of COVID-19 is assessed according to the SARS-CoV-2 diagnosis and treatment guidelines issued by the National Health Commission of China (Chen et al., Detectable serum SARS-CoV-2 viral load (RNA aemia) is closely associated with drastically elevated interleukin 6 (IL-6) levels in critically ill COVID-19 patients. medRxiv 2020.02.29.20029520; Liu et al., The potential role of IL-6 in monitoring severe case of coronavirus disease 2019. medRxiv 2020.03.01.20029769; Zhang et al., Allergy. 2020 Jul; 75 (7): 1730-1741).

[0056] In one embodiment, the method is based on the Interim clinical guidance for patients suspected of / confirmed with COVID-19 in Belgium (March 19, 2020; version 4) of the Belgium National Public Health Institute (Sciensano), retrieved from https: / / epidemio.wiv-isp.be / ID / Documents / Covid19 / COVID-19_

[0057] InterimGuidelines_Treatment_ENG.pdf) to assess the severity of COVID-19.

[0058] In one embodiment, the severity of COVID-19 is assessed according to the World Health Organization (WHO) severity criteria. The WHO COVID-19 severity criteria are as follows:

[0059] - Mild: cases with mild clinical symptoms and no signs of pneumonia in imaging studies.

[0060] -Moderate: Fever and respiratory symptoms with radiologic findings of pneumonia and requiring oxygen (O 2 ): 3L / min <O 2 Cases with blood pressure <5 L / min;

[0061] - Severe: Cases that meet any of the following criteria:

[0062] o Respiratory distress (respiratory rate (RR) ≥ 30 breaths / min);

[0063] o Resting oxygen saturation in ambient air (SpO 2 )≤93%; or SpO 2 ≤ 97% with O 2 >5L / min;

[0064] oThe ratio of arterial oxygen partial pressure to inspired oxygen concentration (PaO 2 / FiO 2 )≦300mmHg (1mmHg=0.133kPa), PaO in high altitude areas (altitude is above 1000 meters above sea level) 2 / FiO 2 The following formula should be used for correction: PaO 2 / F O 2 [multiplied by] [atmospheric pressure (mmHg) / 760]; and / or

[0065] o Chest imaging shows significant lesion progression >50% within 24-48 hours;

[0066] - Critical: Patients who meet any of the following criteria:

[0067] oRespiratory failure and requiring mechanical ventilation;

[0068] oShock; and / or

[0069] oOther organ failure requiring ICU care.

[0070] In one embodiment, the severity and / or progression of COVID-19 is assessed using the WHO 10-point progression scale as shown in Table 1 below (WHO Working Group on the Clinical Characterisation and Management of COVID-19 infection. A minimal common outcome measure set for COVID-19 clinical research. Lancet Infect Dis. 2020 Aug; 20(8): e192-e197. doi: 10.1016 / S1473-3099(20)30483-7).

[0071] Table 1: WHO COVID-19 10-point progression scale

[0072]

[0073]

[0074] In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score range of 2 to 9 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score range of 2 to 5 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score of 2, 3, 4, or 5 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score of 2 or 3 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score range of 4 to 9 on the WHO COVID-19 10-point progression scale (as described in Table 1), preferably in the range of 4 to 6, more preferably 4 or 5. In one embodiment, the subject to be treated according to the present invention has COVID-19 and scores 4, 5 or 6 on the WHO COVID-19 10-point progression scale (as described in Table 1).

[0075] In one embodiment, the subject to be treated according to the present invention has COVID-19 and is hospitalized, but does not require an ICU upon admission, and:

[0076] - A score of 5 on the WHO COVID-19 10-point Progress Scale (as described in Table 1); and

[0077] - Requires more than 3 L / min of oxygen but does not require non-invasive ventilation (NIV) or high flow.

[0078] In one embodiment, the severity and / or progression of COVID-19 is assessed using a modified WHO 7-point progression scale as shown in Table 2 below.

[0079] Table 2: Modified WHO COVID-19 7-point Progress Scale

[0080] Descriptors Fraction No hospitalization, no restriction on activities 1 No hospitalization, limited activities 2 Hospitalized, no need for supplemental oxygen 3 Hospitalized, requiring supplemental oxygen 4 Hospitalization, receiving noninvasive ventilation (NIV) or high-flow oxygen equipment 5 Hospitalization with invasive mechanical ventilation or extracorporeal membrane oxygenation (ECMO) 6 die 7

[0081] In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score ranging from 2 to 6 on the WHO COVID-19 7-point progression scale (as described in Table 2), preferably ranging from 2 to 5. In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score ranging from 3 to 6 on the WHO COVID-19 7-point progression scale (as described in Table 2), preferably ranging from 3 to 5. In one embodiment, the subject to be treated according to the present invention has COVID-19 and has a score of 3, 4 or 5, preferably 4 or 5, on the modified WHO COVID-19 7-point progression scale (as described in Table 2).

[0082] In one embodiment, COVID-19 is mild to moderate COVID-19. Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in preventing and / or treating mild to moderate COVID-19 in a subject in need thereof.

[0083] In one embodiment, mild to moderate COVID-19 is defined as a score ranging from 1 to 5 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, mild to moderate COVID-19 is defined as a score of 1, 2, 3, 4, or 5 on the WHO COVID-19 10-point progression scale (as described in Table 1).

[0084] In one embodiment, mild to moderate COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection associated with at least one of the following clinical symptoms: fever, respiratory symptoms (e.g., cough, shortness of breath, and / or chest tightness), and imaging findings of pneumonia.

[0085] In one embodiment, subjects with mild to moderate COVID-19 are not hospitalized. In one embodiment, subjects with mild to moderate COVID-19 are hospitalized. In one embodiment, subjects with mild to moderate COVID-19 are hospitalized but do not need to be admitted to an intensive care unit (ICU).

[0086] In one embodiment, a subject with mild to moderate COVID-19 as described above requires oxygen therapy. In one embodiment, a subject with mild to moderate COVID-19 as described above requires non-invasive ventilation (NIV).

[0087] In one embodiment, mild COVID-19 is defined as not requiring oxygen (O 2 ) or laboratory-confirmed SARS-CoV-2 infection without evidence of pneumonia.

[0088] In one embodiment, subjects with mild COVID-19 are not hospitalized. In one embodiment, subjects with mild COVID-19 are hospitalized. In one embodiment, subjects with mild COVID-19 are hospitalized but do not require ICU admission.

[0089] In one embodiment, mild COVID-19 is defined as a score ranging from 1 to 3 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, mild COVID-19 is defined as a score of 1, 2, or 3 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, mild COVID-19 is defined as a score of 4 or 5 on the WHO COVID-19 10-point progression scale (as described in Table 1).

[0090] In one embodiment, mild COVID-19 is defined as COVID-19 that requires hospitalization but does not require oxygen therapy. In one embodiment, mild COVID-19 is defined as COVID-19 that requires hospitalization and requires oxygen therapy via a mask or nasal prongs.

[0091] In one embodiment, moderate COVID-19 is defined as a score of 4 or 5 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, moderate COVID-19 is defined as a score of 5 on the WHO COVID-19 10-point progression scale (as described in Table 1), requiring more than 3 L / min of oxygen, but not requiring non-invasive ventilation (NIV) or high flow.

[0092] In one embodiment, moderate COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection associated with the following clinical symptoms: fever, respiratory symptoms (e.g., dry cough, shortness of breath, and / or chest tightness), and radiographic findings of pneumonia.

[0093] In one embodiment, subjects with moderate COVID-19 are not hospitalized. In one embodiment, subjects with moderate COVID-19 are hospitalized. In one embodiment, subjects with moderate COVID-19 are hospitalized but do not require ICU admission.

[0094] In one embodiment, a subject with moderate COVID-19 is hospitalized but does not require ICU admission, wherein moderate COVID-19 is defined as a score of 5 on the WHO COVID-19 10-point progression scale (as described in Table 1) and requires more than 3 L / min of oxygen but does not require non-invasive ventilation (NIV) or high flow.

[0095] In one embodiment, a subject with moderate COVID-19 as described above requires oxygen therapy. In one embodiment, a subject with moderate COVID-19 as described above requires NIV.

[0096] In one embodiment, COVID-19 is severe COVID-19. Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in preventing and / or treating severe COVID-19 in a subject in need thereof.

[0097] In one embodiment, severe COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection with at least one of the following:

[0098] - Respiratory distress with respiratory rate (or respiratory rate (RR)) ≥ 30 / min;

[0099] - Pulse oxygen saturation ≤ 93% at rest; and / or

[0100] - Oxygenation index (ratio of arterial oxygen partial pressure / inspired oxygen concentration (PaO 2 / FiO 2 ))≤300mmHg.

[0101] In one embodiment, severe COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection with at least one of the following:

[0102] - Tachypnea, respiratory rate (RR) ≥ 30 times / min;

[0103] - Blood oxygen saturation less than 93% at rest;

[0104] -PaO 2 / FiO 2 ≤300 mmHg; and / or

[0105] - Radiographic evaluation demonstrates progression of lung lesions greater than 50% within 24 to 48 hours.

[0106] In one embodiment, severe COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection with at least one of the following:

[0107] - Respiratory rate ≥30 / min (adults); ≥40 / min (children <5 years old);

[0108] - Blood oxygen saturation ≤ 93%:

[0109] -PaO 2 / FiO 2 ≤300 mmHg; and / or

[0110] - Pulmonary infiltrates > 50% of lung fields within 24-48 hours.

[0111] In one embodiment, severe COVID-19 is defined as a score ranging from 6 to 9 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, severe COVID-19 is defined as a score of 6, 7, 8, or 9 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, severe COVID-19 is defined as a score of 6 on the WHO COVID-19 10-point progression scale (as described in Table 1).

[0112] In one embodiment, severe COVID-19 is defined as COVID-19 requiring hospitalization and NIV or high-flow oxygen therapy.

[0113] In one embodiment, the subject with severe COVID-19 is hospitalized. In one embodiment, the subject with severe COVID-19 is hospitalized but does not need to be admitted to the ICU. In one embodiment, the subject with severe COVID-19 needs to be admitted to the ICU.

[0114] In one embodiment, a subject with severe COVID-19 as described above requires oxygen therapy. In one embodiment, a subject with severe COVID-19 as described above requires NIV.

[0115] In one embodiment, COVID-19 is critical COVID-19. Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in preventing and / or treating critical COVID-19 in a subject in need thereof.

[0116] In one embodiment, critical COVID-19 is defined as laboratory-confirmed SARS-CoV-2 infection with at least one of the following in addition to the criteria / criteria present in severe COVID-19:

[0117] - Respiratory failure requiring mechanical ventilation;

[0118] - shock (septic shock); and / or

[0119] - Multi-organ failure (extra-pulmonary organ failure) requiring admission to the intensive care unit (ICU).

[0120] In one embodiment, critical COVID-19 is defined as a score in the range of 7 to 9 on the WHO COVID-19 10-point progression scale (as described in Table 1). In one embodiment, critical COVID-19 is defined as a score of 7, 8, or 9 on the WHO COVID-19 10-point progression scale (as described in Table 1).

[0121] In one embodiment, critical COVID-19 is defined as requiring hospitalization, intubation and mechanical ventilation, PaO 2 / FIO 2 ≥ 150 mm Hg or SpO 2 / FIO 2 ) ≥ 200 mmHg for COVID-19.

[0122] In one embodiment, critical COVID-19 is defined as COVID-19 requiring hospitalization and one of the following:

[0123] -Mechanical ventilation (PaO2 / FIO 2 <150 mmHg or SpO 2 / FIO 2 <200 mmHg); or

[0124] - Vasopressors (norepinephrine > 0.3 mcg / kg / min).

[0125] In one embodiment, critical COVID-19 is defined as COVID-19 requiring hospitalization and one of the following:

[0126] - Mechanical ventilation (PaO2 / FIO2 <150 mmHg) and vasopressors (norepinephrine >0.3 mcg / kg / min);

[0127] - dialysis; or

[0128] -ECMO.

[0129] In one embodiment, the subject with critical COVID-19 is hospitalized. In one embodiment, the subject with critical COVID-19 requires admission to the ICU.

[0130] In one embodiment, a subject with critical COVID-19 as described above requires oxygen therapy. In one embodiment, a subject with critical COVID-19 as described above requires NIV. In one embodiment, a subject with critical COVID-19 as described above requires invasive ventilation, such as intubation and mechanical ventilation. In one embodiment, a subject with critical COVID-19 as described above requires vasopressor therapy (e.g., phenylephrine, norepinephrine, epinephrine, vasopressin, and / or dopamine).

[0131] In one embodiment, COVID-19 may cause COVID-19 associated pneumonia (also referred to as COVID-19 pneumonia). Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in preventing and / or treating COVID-19 associated pneumonia in a subject in need thereof.

[0132] In one embodiment, COVID-19 related pneumonia affects both lungs. In one embodiment, COVID-19 related pneumonia appears as blurred patches on lung scans (e.g., computed tomography (CT) scans), particularly blurred patches clustered at the outer edges of the lungs. In one embodiment, COVID-19 related pneumonia appears as abnormal ground-glass opacity radiological findings or mixed pattern radiological findings (a combination of consolidation, ground-glass opacity, and reticular opacity in the presence of architectural distortion) on lung scans.

[0133] In one embodiment, COVID-19 may lead to COVID-19-related acute respiratory distress syndrome (ARDS) (also referred to as COVID-19ARDS). Therefore, in one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described herein, in particular masitinib, or a pharmaceutically acceptable salt or solvate thereof, for use in preventing and / or treating COVID-19-related ARDS in a subject in need thereof.

[0134] In one embodiment, ARDS is defined as a form of acute lung injury (ALI) and occurs as a result of severe lung damage, causing uneven alveolar damage throughout the lung.

[0135] In one embodiment, the subject is male. In one embodiment, the subject is female.

[0136] In one embodiment, the age of the subject is less than 80, 75, 70, 65 or 60 years old. In one embodiment, the subject is under 80 years old. In one embodiment, the subject is under 60 years old. In one embodiment, the age of the subject is greater than 40 years old. In one embodiment, the age of the subject is greater than 60, 65, 70 or 75 years old. In one embodiment, the age of the subject is greater than 60, 65, 70 or 75 years old and less than 80 years old. In one embodiment, the subject is over 60 years old. In one embodiment, the subject is over 60 years old and less than 80 years old. In one embodiment, the subject is over 80 years old. In one embodiment, the age of the subject is greater than 80 years old. In one embodiment, the subject lives in a nursing home or long-term care facility.

[0137] In one embodiment, the subject is not hospitalized. In one embodiment, the subject is hospitalized. In one embodiment, the subject is hospitalized but does not need to be admitted to an intensive care unit (ICU). In one embodiment, the subject is hospitalized and needs to be admitted to an intensive care unit (ICU). In one embodiment, the subject does not need oxygen therapy. In one embodiment, the subject needs oxygen therapy. In one embodiment, the subject needs NIV. In one embodiment, the subject needs invasive ventilation, such as intubation and mechanical ventilation.

[0138] In one embodiment, the subject to be treated according to the present invention has not received or is not currently receiving any other active agent. In one embodiment, the subject to be treated according to the present invention has not received or is not currently receiving any other antiviral agent. Therefore, in one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is administered as a first-line treatment. In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is administered as the only active agent.

[0139] In one embodiment, the subject has an interleukin 6 (IL6) plasma level above 20 pg / mL, particularly an interleukin 6 (IL6) plasma level at baseline. In one embodiment, the subject has an interleukin 6 (IL6) plasma level equal to or below 20 pg / mL, particularly an interleukin 6 (IL6) plasma level at baseline.

[0140] In one embodiment, the subject is at risk of developing a disease caused by a nested virus infection or a picornavirus infection, preferably a coronavirus infection, such as COVID-19 caused by a SARS-CoV-2 infection. In one embodiment, the subject is at risk of developing a severe or critical form of the disease caused by a coronavirus infection, such as COVID-19 caused by a SARS-CoV-2 infection. In one embodiment, the subject is at risk of developing severe or critical COVID-19 as described above. In one embodiment, the subject has COVID-19 and is at risk of developing at least one of the following: pneumonia, acute respiratory distress syndrome (ARDS), sepsis, septic shock, altered consciousness, and / or multiple organ failure.

[0141] In one embodiment, the subject exhibits at least one risk factor that may lead to an increased risk of developing a disease caused by a nested virus infection or a picornavirus infection, preferably a coronavirus infection, such as COVID-19 caused by a SARS-CoV-2 infection. In one embodiment, the subject exhibits at least one risk factor that may lead to an increased risk of developing a severe or critical form of the disease caused by a coronavirus infection, such as COVID-19 caused by a SARS-CoV-2 infection. In one embodiment, the subject exhibits at least one risk factor that may lead to an increased risk of developing a severe or critical form of COVID-19 as described above.

[0142] As used herein, "risk factor" refers to a pre-existing disease, condition, habit or behavior that may lead to an increased risk of developing a disease caused by a nested virus infection or a picornavirus infection, preferably a coronavirus infection, such as COVID-19 caused by SARS-CoV-2 infection. In one embodiment, a "risk factor" refers to a pre-existing disease, condition, habit or behavior that may lead to an increased risk of developing a severe or critical form of a disease caused by a coronavirus infection, such as COVID-19 caused by SARS-CoV-2 infection.

[0143] In one embodiment, the subject exhibits at least one risk factor selected from the group comprising or consisting of: active chemotherapy or radical radiotherapy for lung cancer, active smoking, acute kidney injury, asthma, atopy, autoimmune diseases or conditions, autoinflammatory diseases or conditions, bone marrow or stem cell transplantation within the past 6 months, bronchial hyperresponsiveness, blood or bone marrow cancer (such as leukemia, lymphoma or myeloma) at any stage of treatment, cardiovascular disease or condition, chronic bronchitis, chronic kidney disease, chronic obstructive pulmonary disease (COPD), long-term passive smoking (also known as environmental exposure to smoking), cystic fibrosis, diabetes, emphysema, blood disease, hypertension, immunodeficiency, immunosuppressive therapy (especially immunosuppressive therapy sufficient to significantly increase the risk of infection), immunotherapy or antibody treatment of cancer, HIV (human immunodeficiency virus) infection, lung cancer, obesity, pregnant women (especially pregnant women with severe heart disease (whether congenital or acquired)), pulmonary hypertension (pulmonary hypertension), rare diseases and inborn errors of metabolism that significantly increase the risk of infection (e.g., severe combined immunodeficiency or homozygous sickle cell), reactive airway disease, recipients of solid organ transplants, severe respiratory disease, sickle cell disease, solid cancers, and targeted cancer therapies that may affect the immune system (e.g., protein kinase inhibitors or PARP inhibitors).

[0144] In one embodiment, the subject suffers from at least one comorbidity.

[0145] As used herein, "comorbidity" refers to a disease or condition that coexists with a nested virus infection or a picornavirus infection, preferably a coronavirus infection, such as SARS-CoV-2 infection that causes COVID-19, in a subject to be treated according to the present invention. Examples of comorbidities that may coexist with a nested virus infection or a picornavirus infection, preferably a coronavirus infection, such as SARS-CoV-2 infection that causes COVID-19, in a subject to be treated according to the present invention include, but are not limited to, acute kidney injury, asthma, atopy, autoimmune diseases or conditions, autoinflammatory diseases or conditions, bone marrow or stem cell transplantation within the past 6 months, bronchial hyperresponsiveness, blood or bone marrow cancers (such as leukemia, lymphoma or myeloma) at any stage of treatment, cardiovascular diseases or conditions, chronic bronchitis, chronic kidney disease, chronic obstructive pulmonary disease (COPD), cystic fibrosis, diabetes, emphysema, blood diseases, hypertension, immunodeficiency, HIV infection, lung cancer, obesity, pulmonary hypertension, rare diseases and inborn errors of metabolism (which significantly increase the risk of infection (such as severe combined immunodeficiency or homozygous sickle cell)), reactive airway disease, recipients of solid organ transplants, severe respiratory diseases, sickle cell disease and solid cancers.

[0146] In one embodiment, the subject exhibits at least one comorbidity selected from the group comprising or consisting of acute kidney injury, asthma, atopy, autoimmune diseases or conditions, autoinflammatory diseases or conditions, bone marrow or stem cell transplantation within the past 6 months, bronchial hyperresponsiveness, blood or bone marrow cancer (such as leukemia, lymphoma or myeloma) at any stage of treatment, cardiovascular disease or condition, chronic bronchitis, chronic kidney disease, chronic obstructive pulmonary disease (COPD), cystic fibrosis, diabetes, emphysema, blood disease, hypertension, immunodeficiency, HIV infection, lung cancer, obesity, pulmonary hypertension, rare diseases and inborn errors of metabolism (which significantly increase the risk of infection (such as severe combined immunodeficiency or homozygous sickle cell)), reactive airway disease, recipients of solid organ transplants, severe respiratory diseases, sickle cell disease and solid cancers.

[0147] In one embodiment, the subject has sickle cell disease.

[0148] In one embodiment, the subject is at least one of the following: an active smoker or a long-term passive smoker (that is, the subject is exposed to a smoking environment), immunocompromised, pregnant (especially with severe heart disease (whether congenital or acquired)), receiving active chemotherapy or radical radiotherapy for lung cancer, receiving immunosuppressive therapy (especially immunosuppressive therapy sufficient to significantly increase the risk of infection), receiving cancer immunotherapy or antibody therapy, receiving targeted cancer therapy that can affect the immune system (such as protein kinase inhibitors or PARP inhibitors).

[0149] In the present invention, 2-aminoarylthiazole derivatives refer to compounds characterized by the presence of a thiazole group substituted with a secondary or tertiary amine at the 2-position (i.e., between the heterocyclic nitrogen atom and the sulfur atom), wherein the nitrogen atom of the amine is substituted with at least one aryl group.

[0150] According to one embodiment, the aryl group is substituted with an arylamide group (ie -NH-CO-aryl).

[0151] In one embodiment, the 2-aminoarylthiazole derivatives of the present invention have the following formula (I):

[0152]

[0153] in:

[0154] -R 1 and R 2 are independently selected from hydrogen, halogen, (C 1 -C 10 ) alkyl, (C 3 -C 10 )cycloalkyl, trifluoromethyl, alkoxy, cyano, dialkylamino, solubilizing groups and (C 1 -C 10 )alkyl;

[0155] -m is 0-5;

[0156] -n is 0-4;

[0157] -R 3 is one of the following:

[0158] (i) aryl (e.g. phenyl), which is optionally substituted by one or more substituents such as halogen, (C 1 -C 10 ) alkyl, trifluoromethyl, cyano and alkoxy substituted;

[0159] (ii) heteroaryl (e.g. 2, 3 or 4-pyridyl), which is optionally substituted by one or more substituents such as halogen, (C 1 -C 10 ) alkyl, trifluoromethyl and alkoxy substituted;

[0160] (iii) a five-membered aromatic heterocyclic group (e.g., 2-thienyl, 3-thienyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), which is optionally substituted by one or more substituents such as halogen, (C 1 -C 10 ) alkyl, trifluoromethyl and alkoxy substituted.

[0161] In one embodiment, R of formula (I) 1 and R 2 are independently selected from hydrogen, halogen, (C 1 -C 10 ) alkyl, (C 3 -C 10 ) cycloalkyl, trifluoromethyl, alkoxy, cyano, dialkylamino and solubilizing groups.

[0162] Therefore, in one embodiment, the 2-aminoarylthiazole derivative of the present invention or a pharmaceutically acceptable salt or solvate thereof is a 2-aminoarylthiazole derivative of formula (I) as described above or a pharmaceutically acceptable salt or solvate thereof.

[0163] In one embodiment, the 2-aminoarylthiazole derivatives of the present invention have the following formula (II):

[0164]

[0165] in:

[0166] -R 1 are independently selected from hydrogen, halogen, (C 1 -C 10 ) alkyl, (C 3 -C 10 )cycloalkyl, trifluoromethyl, alkoxy, amino, alkylamino, dialkylamino, solubilizing groups, and (C 1 -C 10 )alkyl; and

[0167] -m is 0-5.

[0168] In one embodiment, R of formula (II) 1 are independently selected from hydrogen, halogen, (C 1 -C 10 ) alkyl, (C 3 -C 10 ) cycloalkyl, trifluoromethyl, alkoxy, amino, alkylamino, dialkylamino and solubilizing groups.

[0169] In one embodiment, R of formula (II) 1In one embodiment, R of formula (II) 1 is substituted with a solubilizing group (C 1 -C 10 )alkyl.

[0170] In one embodiment, R of formula (II) 1 is (C1-C10)alkyl-(C2-C11)heterocycloalkyl-(C1-C10)alkyl-. In one embodiment, R of formula (II) 1 Yes (C 1 -C 4 )alkyl-(C 2 -C 11 )heterocycloalkyl-(C 1 -C 10 )alkyl-, preferably (C 1 -C 2 )alkyl-(C 2 -C 11 )heterocycloalkyl-(C 1 -C 10) In one embodiment, R of formula (II) 1 Yes (C 1 -C 10 )alkyl-(C 2 -C 11 )heterocycloalkyl-(C 1 -C 4 )alkyl-, preferably (C 1 -C 10 )alkyl-(C 2 -C 11 )heterocycloalkyl-(C 1 -C 2 )alkyl-. In one embodiment, R of formula (II) 1 Yes (C 1 -C 10 )alkyl-(C 2 -C 6 )heterocycloalkyl-(C 1 -C 10 )alkyl-, preferably (C 1 -C 10 )alkyl-(C 4 )heterocycloalkyl-(C 1 -C 10 )alkyl-. In one embodiment, R of formula (II) 1 Yes (C 1 -C 4 )alkyl-(C 2 -C 6 )heterocycloalkyl-(C 1 -C4 )alkyl-, preferably (C 1 -C 2 )alkyl-(C 4 )heterocycloalkyl-(C 1 -C 2 )alkyl-. In one embodiment, R of formula (II) 1 Yes (C 1 -C 4 )alkyl-piperazinyl-(C 1 -C 4 )alkyl-, preferably (C 1 -C 2 )alkyl-piperazinyl-(C 1 -C 2 )alkyl-. In one embodiment, R of formula (II) 1 is methylpiperazinyl-(C 1 -C 2 )alkyl-, preferably methylpiperazinyl-methyl-, more preferably 4-methylpiperazinyl-methyl-.

[0171] Therefore, in one embodiment, the 2-aminoarylthiazole derivative of the present invention or a pharmaceutically acceptable salt or solvate thereof is a 2-aminoarylthiazole derivative of formula (II) as described above or a pharmaceutically acceptable salt or solvate thereof.

[0172] As used herein, the term "aryl" refers to a polyunsaturated aromatic hydrocarbon group having a single aromatic ring (i.e., phenyl) or multiple aromatic rings fused together (e.g., naphthyl) or covalently linked, typically containing 5 to 12 atoms; preferably 6 to 10, wherein at least one ring is aromatic. The aromatic ring may optionally include one to two additional rings (cycloalkyl, heterocyclyl, or heteroaryl) fused thereto. Aryl is also intended to include partially hydrogenated derivatives of the carbocyclic ring systems listed herein. Examples of suitable aryl groups include, but are not limited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fused carbocyclic moieties, such as 5,6,7,8-tetrahydronaphthyl. Aryl may be unsubstituted or substituted with one or more substituents. In one embodiment, aryl is a monocyclic ring wherein the ring contains 6 carbon atoms, referred to herein as "(C 6 )aryl".

[0173] As used herein, the term "alkyl" refers to a saturated straight or branched non-cyclic hydrocarbon having 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms. Representative saturated straight chain alkyls include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. Saturated branched alkyls include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl, 2,2-di ... The alkyl group included in the compounds of the present invention can be optionally substituted with one or more substituents.

[0174] As used herein, the term "alkoxy" refers to an alkyl group connected to another moiety through an oxygen atom. Examples of alkoxy groups include, but are not limited to, methoxy, isopropoxy, ethoxy, tert-butoxy. Alkoxy groups may be optionally substituted with one or more substituents.

[0175] As used herein, the term "cycloalkyl" refers to a saturated cyclic alkyl group having 3 to 10 carbon atoms. Representative cycloalkyl groups include cyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl and cyclodecyl. Cycloalkyl groups may be optionally substituted with one or more substituents.

[0176] As used herein, the term "halogen" refers to -F, -Cl, -Br or -I.

[0177] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic heteroaromatic ring comprising carbon atom ring members and one or more heteroatom ring members (e.g., oxygen, sulfur, or nitrogen). Typically, a heteroaryl has 1 to about 5 heteroatom ring members and 1 to about 14 carbon atom ring members. Representative heteroaryls include, but are not limited to, pyridyl, 1-oxo-pyridyl, furanyl, benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl, pyrrolyl, oxazolyl, imidazolyl, thiazolyl, isoxazolyl, quinolyl, pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl , triazolyl, thiadiazolyl, isoquinolyl, indazolyl, benzoxazolyl, benzofuranyl, indolizinyl, imidazopyridinyl, tetrazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl, azaindolyl, imidazopyridinyl, quinazolinyl, purinyl, pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl, imidazo[1,2-a]pyridinyl and benzo(b)thienyl. The heteroatoms may be substituted by protecting groups known to those of ordinary skill in the art, for example, the hydrogen on nitrogen may be substituted by tert-butyloxycarbonyl. The heteroaryl may optionally be substituted by one or more substituents. In addition, the nitrogen or sulfur heteroatom ring members may be oxidized. In one embodiment, the heteroaromatic ring is selected from a 5-8 membered monocyclic heteroaryl ring. The point of attachment of a heteroaromatic or heteroaryl ring to another group can be at a carbon atom or a heteroatom of the heteroaromatic or heteroaryl ring.

[0178] As used herein, the term "heterocycle" refers collectively to heterocycloalkyl and heteroaryl groups.

[0179] As used herein, the term "heterocycloalkyl" refers to a monocyclic or polycyclic group having at least one heteroatom selected from O, N or S and having 2-11 carbon atoms, which may be saturated or unsaturated, but not aromatic. Examples of heterocycloalkyl include, but are not limited to, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 4-piperidonyl, pyrrolidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyridinyl, tetrahydrothiopyran ... The term "heterocycloalkyl" refers to a group consisting of 3 to 7 members, 1,3-dioxolane, 1,3-dioxolane, 1,3-dioxolane, 1,4-dihydrofuranyl, 1,4-dihydrofuranyl-2-one, 1,4-dihydrothiophenyl and 1,1-dioxothiophenyl. Typically, monocyclic heterocycloalkyl has 3 to 7 members. Preferred 3 to 7-membered monocyclic heterocycloalkyls are those with 5 or 6 ring atoms. The heteroatoms may be substituted with protecting groups known to those of ordinary skill in the art, for example, the hydrogen on the nitrogen may be substituted with tert-butyloxycarbonyl. In addition, the heterocycloalkyl may be optionally substituted with one or more substituents. In addition, the point of attachment of the heterocycle to another group may be at a carbon atom or a heteroatom of the heterocycle. Only stable isomers of such substituted heterocyclic groups are considered in this definition.

[0180] As used herein, the term "substituent" or "substituted" refers to the replacement of a hydrogen radical on a compound or group by any desired group that is substantially stable to the reaction conditions in unprotected form or when protected with a protecting group. Examples of preferred substituents include, but are not limited to, halogen (chlorine, iodine, bromine or fluorine); alkyl; alkenyl; alkynyl; hydroxyl; alkoxy; nitro; thiol; thioether; imine; cyano; amido; phosphonate (phosphonato); phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; ester; oxygen (-O); haloalkyl (e.g., trifluoromethyl); cycloalkyl, which may be a single ring or a fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), or heterocycloalkyl, which may be wherein the alkylene group is a monocyclic or fused or non-fused polycyclic group (e.g., pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl or thiazinyl), a monocyclic or fused or non-fused polycyclic aryl or heteroaryl group (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolyl, isoquinolyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzimidazolyl, benzothienyl or benzofuranyl); amino (primary, secondary or tertiary); CO 2 CH 3 ;CONH 2 ;OCH 2 CONH 2 NH 2 ;SO 2 NH 2 ;OCHF 2 CF 3 ;OCF 3 ; and such moieties may also be optionally fused ring structures or bridges such as -OCH 2 O-substituted. These substituents may be optionally further substituted by substituents selected from these groups. In certain embodiments, the term "substituent" or the adjective "substituted" refers to a substituent selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, haloalkyl, -C(O)NR 11 R 12 、-NR 13 C(O)R 14 , halogen, -OR 13 , cyano, nitro, haloalkoxy, -C(O)R 13 、-NR 11 R 12 、-SR 13 、-C(O)OR 13 、-OC(O)R 13 、-NR 13 C(O)NR 11 R12 、-OC(O)NR 11 R 12 、-NR 13 C(O)OR 14 、-S(O)rR 13 、-NR 13 S(O)r 14 、-OS(O)rR 14 、S(O)rNR 11 R 12 , -O, -S, and -NR 13 , where r is 1 or 2; R 11 and R 12 is independently for each occurrence H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl; or R 11 and R 12 together with the nitrogen to which they are attached are optionally substituted heterocycloalkyl or optionally substituted heteroaryl; and R 13 and R 14 is independently for each occurrence H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl. In certain embodiments, the term "substituent" or the adjective "substituted" refers to a solubilizing group.

[0181] As used herein, the term "solubilizing group" refers to any group that can be substantially ionized and enables a compound to be soluble in a desired solvent such as water or an aqueous solvent ("water-solubilizing group"). In addition, a solubilizing group can be a group that increases the lipophilicity of a compound or complex. In one embodiment, the solubilizing group is selected from an alkyl group substituted with one or more heteroatoms such as N, O, S, each heteroatom optionally substituted with an alkyl group independently substituted with an alkoxy, amino, alkylamino, dialkylamino, carboxyl, cyano; or substituted with a cycloheteroalkyl or heteroaryl, or a phosphate, or a sulfate, or a carboxylic acid. In one embodiment, the solubilizing group is one of the following:

[0182] -alkyl, cycloalkyl, aryl, heteroaryl, containing at least one nitrogen or oxygen heteroatom and / or the group is substituted with at least one amino or oxo group (including but not limited to 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 4-piperidonyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydropyranyl, morpholinyl, 1,3-dioxolane, tetrahydrofuranyl and dihydrofuranyl-2-one);

[0183] -amino, which may be a saturated cyclic amino group (including but not limited to piperidinyl, piperazinyl and pyrrolidinyl), which may be substituted by a group consisting of alkyl, alkoxycarbonyl, halogen, haloalkyl, hydroxyalkyl, amino, monoalkylamino, dialkylamino, carbamoyl, monoalkylcarbamoyl and dialkylcarbamoyl (including but not limited to methylpiperidinyl, methylpiperazinyl and methylpyrrolidinyl);

[0184] - one of the structures a) to i) shown below, wherein the wavy lines and arrow lines correspond to the points of attachment to the core structure of the 2-aminoarylthiazole derivative of the present invention (e.g., the 2-aminoarylthiazole derivative of formula (I) or (II)):

[0185]

[0186] In one embodiment, the solubilizing group is a saturated cyclic amino group (including but not limited to piperidinyl, piperazinyl and pyrrolidinyl), which may be substituted with a group consisting of alkyl, alkoxycarbonyl, halogen, haloalkyl, hydroxyalkyl, amino, monoalkylamino, dialkylamino, carbamoyl, monoalkylcarbamoyl and dialkylcarbamoyl (including but not limited to methyl-piperidinyl, methyl-piperazinyl and methyl-pyrrolidinyl).

[0187] In one embodiment, the solubilizing group is structure c) as shown above, wherein the wavy line corresponds to the point of attachment to the core structure of the 2-aminoarylthiazole derivative of the invention (eg, a 2-aminoarylthiazole derivative of formula (I) or (II)).

[0188] As used herein, "pharmaceutically acceptable salts" refer to salts of free acids or free bases that are not biologically undesirable and are generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the free acid with a suitable organic or inorganic base. Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen, phosphate / dihydrogen, phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, toluenesulfonate, trifluoroacetate, and xinofoate. Suitable basic salts are formed from bases that form non-toxic salts. Examples include aluminum, arginine, benzathine penicillin, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, 2-(diethylamino)ethanol, ethanolamine, morpholine, 4-(2-hydroxyethyl)morpholine, and zinc salts. Hemisalts of acids and bases, such as hemisulphate and hemicalcium salts, can also be formed.

[0189] In one embodiment, the pharmaceutically acceptable salt is a pharmaceutically acceptable acid addition salt, for example a pharmaceutically acceptable acid addition salt formed with an inorganic acid, for example hydrochloric acid, sulfuric acid or phosphoric acid, or with a suitable organic carboxylic acid or sulfonic acid, for example an aliphatic mono- or dicarboxylic acid, for example trifluoroacetic acid, acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, malic acid, tartaric acid, citric acid or oxalic acid, or an amino acid, for example arginine or lysine, an aromatic carboxylic acid, for example benzoic acid, 2-phenoxy-benzoic acid, 2-acetoxy-benzoic acid, salicylic acid, 4-aminosalicylic acid, an aromatic-aliphatic carboxylic acid, for example mandelic acid or cinnamic acid, a heteroaromatic carboxylic acid, for example nicotinic acid or isonicotinic acid, an aliphatic sulfonic acid, for example methane-, ethane- or 2-hydroxyethane-sulfonic acid, in particular methanesulfonic acid, or an aromatic sulfonic acid, for example benzene-, p-toluene- or naphthalene-2-sulfonic acid.

[0190] In one embodiment, the pharmaceutically acceptable salt of the 2-aminoarylthiazole derivative of the present invention is a methanesulfonate salt.

[0191] Unless otherwise indicated, the term "mesulphonate" as used herein refers to a salt of methanesulfonic acid with a specified drug substance (e.g., a compound of formula (I) or (II)). The use of mesilate rather than mesylate complies with the INNM (International nonproprietary names modified) published by WHO (e.g., World Health Organization (February 2006). International nonproprietary names revised. INN working document 05.167 / 3.WHO).

[0192] As used herein, "pharmaceutically acceptable solvate" refers to a molecular complex comprising the 2-aminoarylthiazole derivatives of the present invention and a stoichiometric or substoichiometric amount of one or more pharmaceutically acceptable solvent molecules such as ethanol. The term "hydrate" refers to when the solvent is water.

[0193] According to one embodiment, the 2-aminoarylthiazole derivative of the present invention or a pharmaceutically acceptable salt or solvate thereof is masitinib or a pharmaceutically acceptable salt or solvate thereof.

[0194] The chemical name of Masitinib is 4-(4-methylpiperazin-1-ylmethyl]-N-[4-methyl-3-(4-(pyridin-3-ylthiazol-2-ylamino)phenyl]benzamide - CAS number 790299-79-5:

[0195]

[0196] Masitinib was first described in US 7,423,055 and EP 1525200 B1.

[0197] According to one embodiment, the 2-aminoarylthiazole derivative of the present invention or a pharmaceutically acceptable salt or solvate thereof is masitinib mesylate. Therefore, in one embodiment, the pharmaceutically acceptable salt of masitinib as described above is masitinib mesylate. As described above, in other words, the pharmaceutically acceptable salt of masitinib is the mesylate salt of masitinib.

[0198] A detailed procedure for the synthesis of masitinib mesylate is given in WO 2008 / 098949.

[0199] In one embodiment, "masitinib mesylate" refers to the orally bioavailable mesylate salt of masitinib - CAS 1048007-93-7 (MsOH); C 28 H 30 N 6 OS.CH 3 SO 3H;MW 594.76:

[0200]

[0201] According to one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration as the sole active agent (i.e., the only drug that exhibits biological or pharmacological activity). Therefore, according to one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is not used for administration together with another active agent, such as another antiviral agent. In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is not used for administration together with another antiviral agent.

[0202] According to one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration in a therapeutically effective dose.

[0203] In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose of at least about 0.01 mg / kg / day (mg / kg body weight / day), preferably at least about 0.1 mg / kg / day, more preferably at least about 1 mg / kg / day. In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose range of about 1 mg / kg / day to about 500 mg / kg / day, preferably at a dose range of about 1 mg / kg / day to about 200 mg / kg / day.

[0204] In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration in a dosage range of about 1 to about 12 mg / kg / day (mg / kg body weight / day). In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration in a dosage range of about 1.5 to about 7.5 mg / kg / day. In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration in a dosage range of about 3 to about 12 mg / kg / day, preferably about 3 to about 6 mg / kg / day, more preferably about 3 to about 4.5 mg / kg / day.

[0205] In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 mg / kg / day. In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose of about 1.5, 3, 4.5, 6, 7.5, 9, 10.5 or 12 mg / kg / day.

[0206] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose of about 3, 4.5 or 6 mg / kg / day, preferably at a dose of about 3 mg / kg / day or about 4.5 mg / kg / day.

[0207] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at a dose as described above for at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days.

[0208] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, can be dose-escalated in increments of about 1.5 mg / kg / day, to a maximum dose of about 12 mg / kg / day. Toxicity control is performed for each dose increase, and dose escalation is allowed only in the absence of any toxic events.

[0209] In one embodiment, the dose escalation of the 2-aminoaryl thiazole derivative or a pharmaceutically acceptable salt or solvate thereof occurs at any time point after at least 1 day after the initial dose is administered; for example, 1, 2, 3, 4, 5, 6 or 7 days later, preferably 4 days later, more preferably 2 days later. In one embodiment, the dose escalation of the 2-aminoaryl thiazole derivative or a pharmaceutically acceptable salt or solvate thereof occurs at any time point after at least 1 week after the initial dose is administered; for example, 1 week, 2 weeks, 3 weeks or 4 weeks after the initial dose is administered, preferably 1 week later. Toxicity control is performed for each dose increase. Examples of toxicity control include assessments: no suspected severe adverse events were reported during the first 2 days, 4 days or 1 week of treatment with a constant dose of the investigational treatment, no adverse events suspected of leading to treatment interruption, and / or no ongoing suspected adverse events, regardless of their severity, when the dose is increased.

[0210] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at an initial dose of about 3 mg / kg / day during at least 1, 2, 3, 4, 5, 6 or 7 days, preferably during at least 4 days, more preferably during at least 2 days, and then preferably for administration at a dose of about 4.5 mg / kg / day during at least 1, 2, 3, 4, 5 or 6 days. In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration at an initial dose of about 3 mg / kg / day during at least 1 week, at least 2 weeks or at least 3 weeks, and then preferably for administration at a dose of about 4.5 mg / kg / day during at least 1, 2, 3, 4, 5 or 6 days. In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is administered at an initial dose of about 3 mg / kg / day during at least 1, 2, 3, 4, 5, 6 or 7 days, preferably during at least 4 days, more preferably during at least 2 days, then at a dose of about 4.5 mg / kg / day during at least 1, 2, 3, 4, 5, 6 or 7 days, preferably at least 4 days, more preferably at least 2 days, and then at a dose of about 6 mg / kg / day during preferably at least 1, 2, 3, 4, 5 or 6 days. In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is administered at an initial dose of about 3 mg / kg / day for at least 1 week, at least 2 weeks or at least 3 weeks, then at a dose of about 4.5 mg / kg / day for at least 1 week, at least 2 weeks or at least 3 weeks, and then thereafter preferably at a dose of about 6 mg / kg / day for at least 1, 2, 3, 4, 5 or 6 days.

[0211] According to one embodiment, any dosage indicated herein refers to the amount of the active ingredient (also referred to as the active agent) itself, rather than the amount in the form of a pharmaceutically acceptable salt or solvate thereof. Therefore, changes in the composition of the pharmaceutically acceptable salt or solvate of the 2-aminoarylthiazole derivatives (particularly masitinib) of the present invention will not affect the dosage to be administered.

[0212] According to one embodiment, the 2-aminoaryl thiazole derivatives as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, can be administered orally, intravenously, parenterally, topically, by inhalation spray, rectally, nasally or buccally. In one embodiment, the 2-aminoaryl thiazole derivatives as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, can be administered as an oral, sublingual, transdermal, subcutaneous, topical, for absorption through the epithelial layer or mucocutaneous lining, intravenously, intranasally, intraarterially, intramuscularly, intraperitoneally, intrathecally, rectally, vaginally or aerosol formulation.

[0213] Formulations suitable for oral administration may consist of: (a) liquid solutions, such as an effective amount of the compound dissolved in a diluent (e.g., water, saline or orange juice), and including additives such as cyclodextrins (e.g., α-, β- or γ-cyclodextrin, hydroxypropyl cyclodextrin) or polyethylene glycol (e.g., PEG400); (b) capsules, sachets, tablets, lozenges and troches, each of which contains a predetermined amount of active ingredient in solid or granular form; (c) powders; (d) suspensions in appropriate liquids; and (e) suitable emulsions and gels. Liquid formulations may include diluents such as water and alcohols, such as ethanol, benzyl alcohol and polyvinyl alcohol, with or without the addition of pharmaceutically acceptable surfactants, suspending agents or emulsifiers. Capsule forms may be of the ordinary hard or soft shell gelatin type, containing, for example, surfactants, lubricants and inert fillers such as lactose, sucrose, calcium phosphate and corn starch. Tablet forms may include one or more of: lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, gum arabic, gelatin, guar gum, colloidal silicon dioxide, crosslinked sodium carboxymethylcellulose, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid and other excipients, colorants, diluents, buffers, disintegrants, wetting agents, preservatives, flavoring agents and pharmacologically compatible carriers. Lozenge forms may contain the active ingredient in a flavoring agent, which is typically sucrose and gum arabic or tragacanth, and pastilles contain the active ingredient in an inert matrix, such as gelatin and glycerin, or sucrose and gum arabic, emulsions, gels, etc., containing in addition to the active ingredient a carrier known in the art.

[0214] Suitable formulations for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may include suspending agents, solubilizers, thickening agents, stabilizers and preservatives. The active agent can be administered in a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or liquid mixture, including water, saline, dextrose (dextrose) aqueous solution and related sugar solutions, alcohols, such as ethanol, isopropanol or hexadecanol, glycols, such as propylene glycol or polyethylene glycol, glycerol ketal, such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, such as polyethylene glycol (e.g., PEG400), oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides, with or without the addition of pharmaceutically acceptable surfactants, such as fatty acid salts (soap) or detergents, suspending agents, such as pectin, carbomer, methylcellulose, hydroxypropyl methylcellulose or carboxymethylcellulose, or emulsifiers and other pharmaceutical adjuvants. Oils that can be used for parenteral preparations include petroleum, animal oils, vegetable oils or synthetic oils. Specific examples of oils include peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum oil and mineral oil. Suitable fatty acids for parenteral formulations include oleic acid, stearic acid and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters. Suitable fatty acid salts (soaps) for parenteral formulations include fatty alkali metal, ammonium and triethanolamine salts, suitable detergents include (a) cationic detergents, such as dimethyldialkylammonium halides and alkyl pyridinium halides, (b) anionic detergents, such as alkyl, aryl and olefin sulfonates, alkyl, olefin, ether and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents, such as fatty amine oxides, fatty acid alkanolamides and polyoxyethylene-polypropylene copolymers, (d) amphoteric detergents, such as alkyl-β-aminopropionates and 2-alkyl-imidazoline quaternary ammonium salts, and (e) mixtures thereof. Parenteral formulations typically contain about 0.5 to about 25% by weight of an active agent, i.e., a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in solution. Suitable preservatives and buffers may be used in such formulations. In order to minimize or eliminate irritation at the injection site, such compositions may contain one or more nonionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such formulations ranges from about 5 to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and high molecular weight adducts of ethylene oxide with a hydrophobic base, which are formed by condensation of propylene oxide with propylene glycol.Parenteral formulations can be present in unit dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in freeze-dried (lyophilized) conditions, which only require the addition of sterile liquid carriers, such as water, for injection just before use. Ready-to-use injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the aforementioned type.

[0215] The active agent, i.e., a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, can be formulated into an injectable formulation. The requirements for effective drug carriers for injectable compositions are well known to those of ordinary skill in the art. See Pharmaceutics and Pharmacy Practice, JB Lippincott Co., Philadelphia, Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986).

[0216] The composition for topical application is generally in the form of liquid (e.g., mouthwash), cream, paste, lotion, and gel. Topical application includes application to oral mucosa, including oral cavity, oral epithelium, palate, gums, and nasal mucosa. In some embodiments, the composition contains at least one active ingredient and a suitable solvent or carrier. It may also include other components, such as anti-irritants. The carrier may be a liquid, solid, or semisolid. In embodiments, the composition may be an aqueous solution, such as a mouthwash. Alternatively, the composition may be a dispersant, emulsion, gel, lotion, or cream solvent for various components. In one embodiment, the main solvent is water or substantially neutral or has become a substantially neutral biocompatible solvent. The liquid solvent may include other materials, such as buffer, alcohol, glycerol, and mineral oil, and various emulsifiers or dispersants known in the art, to obtain desired pH, consistency, and viscosity. The composition may be produced as a solid, such as a powder or granules. The solid can be applied directly or dissolved in water or a biocompatible solvent prior to use to form a substantially neutral or rendered substantially neutral solution which can then be applied to the target site. In embodiments of the invention, vehicles for topical application to the skin can include water, buffered solutions, various alcohols, glycols such as glycerol, lipid materials such as fatty acids, mineral oils, phosphoglycerides, collagen, gelatin, and silicone-based materials.

[0217] The active agent, i.e., a 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, can be prepared into an aerosol formulation to be administered by inhalation. These aerosol formulations can be placed in pressurized acceptable propellants. Suitable propellants include, for example, fluorinated hydrocarbons (e.g., trichloromonofluoromethane, dichlorodifluoromethane, chlorodifluoromethane, chlorodifluoroethane, dichlorotetrafluoroethane, heptafluoropropane, tetrafluoroethane, difluoroethane), hydrocarbons (e.g., propane, butane, isobutane) or compressed gases (e.g., nitrogen, nitrous oxide, carbon dioxide). They can also be formulated as medicines for non-pressurized formulations, such as in a nebulizer or atomizer.

[0218] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is for oral administration.

[0219] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is for administration at least once a day, preferably twice a day.

[0220] In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration for a period of at least 1, 2, 3, 4, 5 or 6 weeks, preferably for a period of at least 2 weeks. In one embodiment, the 2-aminoaryl thiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for administration for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 days, preferably for a period of at least 15 days.

[0221] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is in a form suitable for oral administration. Examples of forms suitable for oral administration include, but are not limited to, liquids, pastes or solid compositions, more specifically tablets, capsules, pills, liquids, gels, syrups, slurries and suspensions.

[0222] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is for administration as a tablet, preferably as a 100 mg or 200 mg tablet.

[0223] According to one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for combined administration with a quercetin flavonol such as isoquercetin, quercetin or quercetin-3-O-β-D-glucuronide.

[0224] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for combined administration with isoquercetin or quercetin, preferably with isoquercetin.

[0225] Isoquercetin (CAS No. 482-35-9) is also known as quercetin-3-O-pyranoglucoside, quercetin-3-O-glucoside, isoquercitroside, isotrifoliin, trifolin, trifoliin A or isoquercitrin. Its molecular formula is C21H20O12, and its IUPAC (International Union of Pure and Applied Chemistry) name is 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromene-4-one.

[0226] Isoquercetin has the following structural formula:

[0227]

[0228] As used herein, the term "isoquercetin" includes its crystalline solid form, any prodrug, pharmaceutically acceptable salt, hydrate and solvate.

[0229] Isoquercetin is a flavonol belonging to a large class of plant-derived pigments called flavonoids. Flavonoids are the largest group of naturally occurring polyphenolic compounds with a variety of biological activities. Isoquercetin is an orally bioavailable derivative of quercetin.

[0230] Quercetin (CAS No. 117-39-5) is also known as sophoretin, meletin, xanthaurine, quercetol, quercitin, quertine, 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one, 3,3',4',5,7-pentahydroxyflavone or 3,5,7,3',4'-pentahydroxyflavone. Its molecular formula is C15H10O7, and its IUPAC name is 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one.

[0231] Quercetin has the following structural formula:

[0232]

[0233] As used herein, the term "quercetin" includes its crystalline solid form, any prodrug, pharmaceutically acceptable salt, hydrate and solvate.

[0234] Quercetin is an abundant polyphenolic flavonoid that has been isolated from a variety of fruits and plants and exhibits a variety of biological activities.

[0235] Therefore, an object of the present invention is a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in combination with isoquercetin or quercetin, preferably in combination with isoquercetin, for use in treating a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19, in a subject in need thereof as described above.

[0236] In one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in combination with isoquercetin for use in treating coronavirus infection in a subject in need thereof as described above, in particular SARS-CoV-2 infection that causes COVID-19.

[0237] In one embodiment, the present invention relates to a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in combination with isoquercetin for use in preventing and / or treating COVID-19 in a subject in need thereof as described above.

[0238] Therefore, in one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for simultaneous, separate or sequential administration with isoquercetin or quercetin.

[0239] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, is used for combined administration with isoquercetin or quercetin, for example in the form of a combined preparation, pharmaceutical composition or medicament.

[0240] Another object of the present invention is a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and isoquercetin or a combination of quercetin, preferably isoquercetin, for use in treating a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, in particular a SARS-CoV-2 infection that causes COVID-19, in a subject in need thereof as described above.

[0241] In one embodiment, the present invention relates to a combination of masitinib or a pharmaceutically acceptable salt or solvate thereof and isoquercetin for use in treating coronavirus infection, in particular SARS-CoV-2 infection causing COVID-19, in a subject in need thereof as described above.

[0242] In one embodiment, the present invention relates to a combination of masitinib or a pharmaceutically acceptable salt or solvate thereof and isoquercetin for use in preventing and / or treating COVID-19 in a subject in need thereof as described above.

[0243] In one embodiment, the combination of the invention is a simultaneous, separate or sequential combination.In one embodiment, the combination of the invention is a combined preparation, a pharmaceutical composition or a medicament.

[0244] In one embodiment, isoquercetin as described above is used for administration in a dosage range of about 0.25 g / day to about 5 g / day, preferably in a dosage range of about 0.5 g / day to about 2.5 g / day, more preferably in a dosage range of about 1 g / day to about 2 g / day. In one embodiment, isoquercetin as described above is administered in a dosage range of about 0.4 g / day to about 2 g / day.

[0245] In one embodiment, isoquercetin as described above is administered at a dose of about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 g / day. In one embodiment, isoquercetin as described above is administered at a dose of about 1 g / day.

[0246] In one embodiment, isoquercetin as described above may be administered in regularly decreasing doses in increments of about 0.5 g / day.

[0247] In one embodiment, the dose reduction of isoquercetin as described above occurs at any time point after at least 7 days after the initial dose and before 28 days after the initial dose; for example, 7 days, 14 days, or 21 days after the initial dose.

[0248] In one embodiment, isoquercetin as described above is administered at an initial dose of about 2 g / day, followed by administration at a dose of about 1.5 g / day over a period of at least 7, 14 or 21 days.

[0249] In one embodiment, isoquercetin as described above is administered at an initial dose of about 2 g / day for at least 7 days, then at a dose of about 1.5 g / day for at least 7 days, and thereafter at a dose of about 1 g / day.

[0250] According to one embodiment, isoquercetin as described above may be administered orally, intravenously, parenterally, topically, by inhalation spray, rectally, nasally or buccally.

[0251] In one embodiment, isoquercetin as described above is for oral administration.

[0252] In one embodiment, isoquercetin as described above is administered at least once a day, preferably twice a day.

[0253] In one embodiment, isoquercetin as described above is used for administration for a period of at least 1, 2, 3, 4, 5 or 6 weeks, preferably for a period of at least 2 weeks. In one embodiment, isoquercetin as described above is used for administration for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 days, preferably for a period of at least 15 days.

[0254] In one embodiment, isoquercetin as described above is in a form suitable for oral administration. Examples of forms suitable for oral administration are indicated above.

[0255] In one embodiment, isoquercetin as described above is for administration as a capsule, preferably as a 250 mg capsule.

[0256] Another object of the present invention is a kit-of-parts comprising a first part and a second part, wherein the first part comprises a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and the second part comprises isoquercetin or quercetin as described above, preferably isoquercetin.

[0257] In one embodiment, the kit of parts of the present invention comprises a first part comprising masitinib or a pharmaceutically acceptable salt or solvate thereof, and a second part comprising isoquercetin.

[0258] According to one embodiment, the 2-aminoarylthiazole derivative of the present invention, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally together with isoquercetin or quercetin, is administered together with at least one additional pharmaceutically active agent.

[0259] In one embodiment, a 2-aminoarylthiazole derivative as described above, or a pharmaceutically acceptable salt or solvate thereof, is combined with isoquercetin or quercetin, preferably with isoquercetin, for administration together with at least one additional pharmaceutically active agent.

[0260] In one embodiment, the combination of a 2-aminoarylthiazole derivative as described above, or a pharmaceutically acceptable salt or solvate thereof, and isoquercetin or quercetin, preferably with isoquercetin, is for administration together with at least one additional pharmaceutically active agent.

[0261] According to the present invention, the 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally together with isoquercetin or quercetin, may be administered simultaneously, separately or sequentially with the at least one additional pharmaceutically active agent.

[0262] In one embodiment, the 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, optionally together with isoquercetin or quercetin, is used for combined administration with at least one additional pharmaceutically active agent, preferably in the form of a combined preparation, a pharmaceutical composition or a medicament.

[0263] Examples of additional pharmaceutically active agents that can be administered to a subject suffering from a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, particularly a SARS-CoV-2 infection that causes COVID-19, include, but are not limited to, antiviral agents, anti-interleukin 6 (anti-IL6) agents, protease inhibitors, Janus-associated kinase (JAK) inhibitors and other agents, such as BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon or carlimycin.

[0264] In one embodiment, at least one additional pharmaceutically active agent is selected from the group comprising or consisting of: an antiviral agent; an anti-interleukin 6 (anti-IL6) agent; a protease inhibitor; a JAK inhibitor; other agents, such as BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon, carlimycin, angiotensin receptor blockers (ARB), angiotensin converting enzyme inhibitors (ACE-I), losartan or CD24Fc; and any mixtures thereof.

[0265] In one embodiment, at least one additional pharmaceutically active agent is selected from the group comprising or consisting of: an antiviral agent; an anti-interleukin 6 (anti-IL6) agent; a protease inhibitor; a JAK inhibitor; other agents, such as BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon, carlimycin, and any mixture thereof.

[0266] In one embodiment, at least one additional pharmaceutically active agent is selected from the group consisting of: remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir) with or without interferon (e.g., interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b) and pegylated interferon beta-1a), a combination of darunavir and cobicistat (darunavir / cobicistat), oseltamivir, favipiravir, ), hydroxychloroquine, chloroquine, tocilizumab, sarilumab, baricitinib, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon (e.g., interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b) and pegylated interferon beta-1a), carlimycin, angiotensin receptor blockers (ARBs), angiotensin converting enzyme inhibitors (ACE-I), losartan, CD24Fc, and any mixture thereof.

[0267] In one embodiment, at least one additional pharmaceutically active agent is selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir) with or without interferon (e.g., interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b), and pegylated interferon beta-1a), hydroxychloroquine, an anti-IL6 agent (e.g., tocilizumab, siltuximab, salilumab, sirukumab, clazakizumab, or olokizumab), and any mixture thereof.

[0268] In one embodiment, at least one additional pharmaceutically active agent is selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir) with or without interferon (e.g., interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b), and pegylated interferon beta-1a), hydroxychloroquine, and any mixture thereof.

[0269] In one embodiment, at least one additional pharmaceutically active agent is an antiviral agent. Examples of antiviral agents that can be administered to subjects with a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, particularly a SARS-CoV-2 infection causing COVID-19, include, but are not limited to, remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir), chloroquine, hydroxychloroquine, ribavirin, oseltamivir, beclabuvir, saquinavir, umifenovir, favipiravir, leronlimab, a combination of darunavir and cobicistat (darunavir / cobicistat), galidesivir and fabiravir.

[0270] In one embodiment, at least one additional pharmaceutically active agent is an antiviral agent selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir), chloroquine, hydroxychloroquine, ribavirin, oseltamivir, belabuvir, saquinavir, uminavir, favipiravir, lelizumab, a combination of darunavir and cobicistat (darunavir / cobicistat), galisvir, fabiravir, and any mixture thereof.

[0271] In one embodiment, at least one additional pharmaceutically active agent is an antiviral agent selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir), chloroquine, hydroxychloroquine, oseltamivir, favipiravir, a combination of darunavir and cobicistat (darunavir / cobicistat), galisvir, fabiravir, and any mixture thereof.

[0272] In one embodiment, at least one additional pharmaceutically active agent is an antiviral agent selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir), chloroquine, hydroxychloroquine, and any mixture thereof.

[0273] In one embodiment, the at least one additional pharmaceutically active agent is an antiviral agent selected from the group comprising or consisting of remdesivir, a combination of lopinavir and ritonavir (lopinavir / ritonavir), hydroxychloroquine, and any mixture thereof.

[0274] In one embodiment, the at least one additional pharmaceutically active agent is an antiviral agent selected from the group comprising or consisting of remdesivir, hydroxychloroquine, and any mixture thereof.

[0275] In one embodiment, at least one additional pharmaceutically active agent is an anti-IL6 agent. Examples of anti-IL6 agents that can be administered to a subject suffering from a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, in particular a SARS-CoV-2 infection causing COVID-19, include, but are not limited to, tocilizumab, siltuximab, salizumab, salizumab, clazakizumab, and olotuzumab.

[0276] In one embodiment, the at least one additional pharmaceutically active agent is an anti-IL6 agent selected from the group comprising or consisting of tocilizumab, cetuximab, thalirumab, sirukumab, clazakizumab, olotuzumab, and any mixture thereof.

[0277] In one embodiment, the at least one additional pharmaceutically active agent is tocilizumab.

[0278] In one embodiment, at least one additional pharmaceutically active agent is a protease inhibitor. Examples of protease inhibitors that can be administered to subjects suffering from a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, in particular a SARS-CoV-2 infection causing COVID-19, include, but are not limited to, simeprevir and camostat mesylate.

[0279] In one embodiment, the at least one additional pharmaceutically active agent is a protease inhibitor selected from the group comprising or consisting of simiprevir, camostat mesylate, and any mixtures thereof.

[0280] In one embodiment, at least one additional pharmaceutically active agent is a JAK inhibitor. Examples of JAK inhibitors that can be administered to subjects suffering from a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, in particular a SARS-CoV-2 infection causing COVID-19, include, but are not limited to, baricitinib, fedratinib, and ruxolitinib.

[0281] In one embodiment, the at least one additional pharmaceutically active agent is a JAK inhibitor selected from the group comprising or consisting of baricitinib, fumaratetinib, ruxolitinib, and any mixtures thereof.

[0282] Other agents that can be administered to a subject suffering from a nested virus infection or a picornavirus infection as described above, preferably a coronavirus infection, particularly a SARS-CoV-2 infection that causes COVID-19, include, but are not limited to, BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon (such as interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b) and pegylated interferon beta-1a), carlimycin, angiotensin receptor blockers (ARBs), angiotensin converting enzyme inhibitors (ACE-I), losartan, bevacizumab, CD24Fc.

[0283] In one embodiment, at least one additional pharmaceutically active agent is selected from the group comprising or consisting of BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon (e.g., interferon beta-1a (IFN-β-1a), interferon beta-1b (IFN-β-1b) and pegylated interferon beta-1a), carlimycin, angiotensin receptor blockers (ARBs), angiotensin converting enzyme inhibitors (ACE-I), losartan, bevacizumab, CD24Fc, and any mixtures thereof.

[0284] Another object of the present invention is a method for treating a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above, in a subject in need thereof, comprising or consisting of the following steps: administering to the subject a 2-aminoarylthiazole derivative as described above, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof.

[0285] In one embodiment, the method of the present invention comprises or consists of the following steps: administering a pharmaceutical composition as described herein, comprising, consisting essentially of, or consisting of: a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

[0286] In one embodiment, the method of the present invention comprises or consists of administering a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in combination with isoquercetin or quercetin, preferably isoquercetin.

[0287] In one embodiment, the methods of the present invention comprise administering at least one additional pharmaceutically active agent as described above.

[0288] In one embodiment, the methods of the present invention are used to treat SARS-CoV-2 infection causing COVID-19 as described above. In one embodiment, the methods of the present invention are used to prevent and / or treat COVID-19 related pneumonia and / or COVID-19 related acute respiratory distress syndrome (ARDS) in a subject in need thereof as described above.

[0289] Another object of the present invention is a pharmaceutical composition for treating a nested virus infection or a picornavirus infection as described above in a subject in need thereof, wherein the pharmaceutical composition comprises, consists essentially of or consists of: a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient. According to one embodiment, the present invention relates to a pharmaceutical composition for treating a coronavirus infection, in particular an infection with SARS-CoV-2 causing COVID-19, in a subject in need thereof, wherein the pharmaceutical composition comprises, consists essentially of or consists of: a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

[0290] Pharmaceutically acceptable excipients, such as solvents, adjuvants, carriers or diluents, are well known to those skilled in the art and are readily available to the public. Typically, a pharmaceutically acceptable excipient is an excipient that is chemically inert to the active compound (also referred to as an active agent or active ingredient) and has no harmful side effects or toxicity under the conditions of use.

[0291] In one embodiment, the pharmaceutical composition comprises, consists essentially of, or consists of masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition consists of masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

[0292] In one embodiment, the pharmaceutical composition of the present invention further comprises isoquercetin or quercetin, preferably isoquercetin. Therefore, in one embodiment, the pharmaceutical composition comprises, consists essentially of, or consists of: masitinib or a pharmaceutically acceptable salt or solvate thereof; isoquercetin or quercetin, preferably isoquercetin; and at least one pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition consists of masitinib or a pharmaceutically acceptable salt or solvate thereof; isoquercetin or quercetin, preferably isoquercetin; and at least one pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition consists of masitinib or a pharmaceutically acceptable salt or solvate thereof, isoquercetin, and at least one pharmaceutically acceptable excipient.

[0293] In one embodiment, the pharmaceutical composition of the present invention is used in combination with isoquercetin or quercetin, preferably isoquercetin, to treat a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above.

[0294] Therefore, another object of the present invention is a pharmaceutical composition as described above for use in the treatment of a nested virus infection or a picornavirus infection, preferably a coronavirus infection, in a subject in need thereof, in combination with isoquercetin or quercetin, preferably isoquercetin, wherein the pharmaceutical composition comprises, consists essentially of or consists of a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

[0295] Another object of the present invention is a pharmaceutical composition in combination with isoquercetin or quercetin, preferably isoquercetin, for use in the treatment of a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above, in a subject in need thereof, wherein the pharmaceutical composition comprises, consists essentially of or consists of a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

[0296] In one embodiment, the pharmaceutical composition of the present invention is used to treat SARS-CoV-2 infection causing COVID-19 as described above. In one embodiment, the pharmaceutical composition of the present invention is used to prevent and / or treat COVID-19 related pneumonia and / or COVID-19 related acute respiratory distress syndrome (ARDS) in a subject in need thereof.

[0297] Another object of the present invention is the use of a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, in the preparation of a medicament for treating a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above, in a subject in need thereof.

[0298] In one embodiment, the present invention relates to the use of a 2-aminoarylthiazole derivative, preferably masitinib, or a pharmaceutically acceptable salt or solvate thereof, and isoquercetin or a combination of quercetin in the preparation of a medicament for treating a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above, in a subject in need thereof.

[0299] In one embodiment, the present invention relates to the use of a 2-aminoarylthiazole derivative as described above or a pharmaceutically acceptable salt or solvate thereof for the preparation of a medicament for treating a nested virus infection or a picornavirus infection, preferably a coronavirus infection, as described above, in a subject in need thereof, wherein the medicament is for administration in combination with isoquercetin or quercetin.

[0300] In one embodiment, the medicament is for administration in combination with at least one additional pharmaceutically active agent as described above.

[0301] In one embodiment, the coronavirus infection is a SARS-CoV-2 infection causing COVID-19 as described above. In one embodiment, the medicament is used to prevent and / or treat COVID-19-related pneumonia and / or COVID-19-related acute respiratory distress syndrome (ARDS) in a subject in need thereof.

[0302] BRIEF DESCRIPTION OF THE DRAWINGS

[0303] Figures 1A-1C is a combination graph illustrating the effects of masitinib, isoquercetin, and the combination of masitinib and isoquercetin on non-senescent cells. Figure 1A The dose-dependent effect of masitinib alone (from 0.1 to 2 μM) on the viability of non-senescent cells is shown. Figure 1B Shown is a dose-dependent effect of isoquercetin alone (from 1 to 20 μM) on the viability of non-senescent cells. Figure 1C The combination of masitinib (from 0.1 to 2 μM) and isoquercetin (from 1 to 20 μM) was shown to have a dose-dependent effect on the viability of non-senescent cells.

[0304] Figures 2A-2C is a combination graph illustrating the effects of masitinib, isoquercetin, and the combination of masitinib and isoquercetin on senescent cells. Figure 2A The dose-dependent effect of masitinib alone (from 0.1 to 2 μM) on the viability of senescent cells is shown. Figure 2B Shown is a dose-dependent effect of isoquercetin alone (from 1 to 20 μM) on senescent cell viability. Figure 2C The dose-dependent effect of the combination of masitinib (from 0.1 to 2 μM) and isoquercetin (from 1 to 20 μM) on the viability of senescent cells was shown.

[0305] Figure 3 is a graph showing the average percentage of OC43 infected cells per well versus increasing concentrations of masitinib. Individual measurements are shown as semi-transparent circles (some circles overlap).

[0306] Figure 4 is a line graph showing that masitinib inhibits OC43 replication in primary human airway epithelial cells with an EC50 of 0.58 μM.

[0307] Figure 5 is a graph showing the results of masitinib treatment of A549 cells overexpressing ACE2, which were pretreated with various concentrations of masitinib for 2 hours, infected with SARS-CoV-2 (MOI 0.5) and incubated for 2 days. Cells were stained for the presence of spike protein and analyzed for the percentage of infected cells. Individual measurements are shown as semi-transparent circles (some circles overlap).

[0308] Figure 6 is a graph showing the effect of masitinib on SARS-CoV-2 progeny production. Cells were treated with 10 μM masitinib for 2 hours, infected with SARS-CoV-2 (MOI=0.5) and cell supernatants were collected 2 days later for titration, n=3. Individual measurements are shown as translucent circles. Masitinib showed a statistically significant reduction in viral titers (p value <0.001, one-tailed t-test, FDR correction).

[0309] Fig. 7A -E is a set of graphs showing the effects of masitinib on the major proteases of SARS-CoV-2 (called 3CLpro, M pro or nsp5). Fig. 7A is a bar graph showing the results of a FlipGFP reporter assay designed to assess the inhibitory effect of masitinib on 3CLpro at a single concentration (10 μM). Individual measurements are shown as circles. The bar graph depicts the mean ± SE. Masitinib treatment completely inhibited 3CLpro activity. Figure 7B is a dose-response curve of masitinib inhibition of 3CLpro obtained using the FlipGFP reporter assay, n = 6. Individual measurements are shown as circles. Figure 7C is a dose-response curve of masitinib inhibition of 3CLpro obtained using a luciferase reporter assay, n = 3. Individual measurements are shown as circles. Fig.7D is a dose response curve of masitinib inhibition of 3CLpro obtained in a cell-free assay using purified 3CLpro and a fluorogenic peptide substrate, n = 3. Individual measurements are shown as circles. Fig. 7E is a line graph showing in vitro characterization of the inhibitory effect of masitinib on 3CL in the presence of different substrate (S) concentrations as indicated. Masitinib is a competitive inhibitor of 3CL activity with a Ki value of 2.58 μM.

[0310] Fig. 8A -B shows the interaction of masitinib with the main proteases of SARS-CoV-2 (called 3CLpro, M pro or nsp5). Fig. 8A Dimer formation, domain structure, and masitinib binding site of SARS-CoV-23CLpro are shown. In monomer A, the inhibitor masitinib is drawn in stick form, binding to the active site between D1 and D2. The sites of the three binding pockets S1, S2, and S4 are labeled. Figure 8B The interaction of masitinib with 3CLpro is shown. The ribbon diagram shows the details of some of the interactions formed between masitinib and 3CLpro at the active site. The key pocket forming or interacting residues of 3CLpro are also presented in stick form with their C atoms. Hydrogen bonds are drawn with dashed lines. The two catalytic residues are marked with asterisks.

[0311] Fig. 9A -B is a set of graphs illustrating the inhibitory effect of masitinib on small RNA viruses. Fig. 9A is a bar graph showing the results of a luciferase reporter assay performed to investigate the ability of masitinib to inhibit the proteolytic activity of picornavirus 3C, derived from coxsackievirus B3 (CVB3). n=6, p value=7X10-6 (one-tailed t-test). Fig. 9B The bar graphs shown show the results after Huh7 cells were treated with 10 μM masitinib for 2 hours, infected with coxsackievirus B3 (CVB3) or human rhinovirus 2, 14 and 16 (HRV2, HRV14, HRV16) at an MOI of 0.01, and supernatants were collected after 24 hours for titration. n=3, p value <0.001 (one-tailed t-test, FDR corrected).

[0312] Fig.10Bar graphs shown demonstrate that masitinib (10 μM) had no significant effect on cells infected with influenza A virus (IAV, Orthomyxoviridae), measles virus (MeV, Paramyxoviridae), lymphocytic choriomeningitis virus (LCMV), and chikungunya virus (CHIKV, Togaviridae). All n=3 except LCMV (n=2). p-value>0.07 (one-tailed t-test, FDR corrected).

[0313] Fig.11A -B shows dot plots showing SARS-CoV-2 viral loads in mouse lungs (11A) and mouse nasal nasal conchae (11B) at 4 and 6 days post-infection. Mice were treated with masitinib (25 or 50 mg / kg, bid, ip) or PBS.

[0314] Fig.12 is a line graph showing the clinical scores of mice 1-6 days after infection. Mice were treated with masitinib (25 or 50 mg / kg, bid, ip) or PBS. Example

[0315] The present invention is further illustrated by the following examples.

[0316] Example 1: In vitro senolytic effects of masitinib and isoquercetin

[0317] Materials and methods

[0318] Material

[0319] BV2 cells are retrovirally immortalized microglia-like cells used as a model of cellular senescence. BV2 cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum (hiFBS) and seeded in 6-well multiwell plates for processing and flow cytometric analysis.

[0320] Masitinib (masitinib mesylate) was obtained from AB Science (Paris, France) and prepared as a 0.1-2 μM solution in DMSO.

[0321] Isoquercetin was prepared into 1-20 μM DMSO solution.

[0322] method

[0323] Aging Model

[0324] BV2 cells were treated with temozolomide (TMZ), an alkylating agent that induces DNA damage, to induce cellular senescence. For senescence induction studies, cells were plated and treated twice with increasing doses (10-150 μM) of TMZ over a 5-hour period every 24 hours. After two consecutive exposures of BV2 cells to TMZ treatment, their proliferation was reduced and the cells developed a characteristic senescent phenotype with increased size and flat granular appearance. The senescent phenotype of BV2 cells after TMZ-induced genotoxicity was further confirmed by measuring the number of cells showing β-gal activity, a well-established marker of cellular senescence. After the second TMZ treatment, flow cytometry analysis was performed by measuring β-galactosidase activity by alkalinization of lysosomes using a 5-dodecanoylaminofluorescein Di-β-D-Galactopyranoside (C12FDG) kit according to the manufacturer's instructions (Thermo Fisher Scientific, #D2893). TMZ induced a dose-dependent increase in the number of β-gal+ cells. Up to 60% of β-gal+ BV2 cells, i.e., senescent cells, were obtained at a TMZ concentration of 100 μM (data not shown).

[0325] Treatment with masitinib, isoquercetin, or a combination of masitinib and isoquercetin

[0326] To analyze the cell viability of non-senescent cells (BV2 cells) and senescent cells (BV2 cells pretreated with TMZ), cells were plated in 96-well multiwell plates over a period of 72 hours. Cells were treated with increasing doses of isoquercetin (1-20 μM in DMSO), masitinib (0.1-2 μM in DMSO), or a combination of isoquercetin and masitinib to investigate any potential synergistic effects. After 48 hours of treatment, cell viability was analyzed by sulforhodamine B (SRB) assay. The optical density (OD) of each well was read at 540 nm in a 96-well plate reader. The OD of the SRB solution is proportional to the number of cells.

[0327] result

[0328] BV2 cells treated with TMZ, a well-suited model for screening anti-aging drugs, were used to evaluate the effects of masitinib alone, isoquercetin alone, or the combination of masitinib and isoquercetin in reducing the viability of senescent cells.

[0329] Therefore, BV2 cells pretreated with TMZ were incubated with masitinib alone, isoquercetin alone, or a combination of masitinib and isoquercetin. As a control, non-senescent proliferating BV2 cells were incubated with masitinib alone, isoquercetin alone, or a combination of masitinib and isoquercetin under the same conditions.

[0330] As shown in Figure 1, when non-senescent proliferating BV2 cells were exposed to masitinib ( Figure 1A ), isoquercetin ( Figure 1B ) or the combination of masitinib and isoquercetin ( Figure 1C ) at 48 h, there was no significant change in cell viability, even at high concentrations of masitinib and / or isoquercetin, as estimated by flow cytometric analysis of live cells stained with sulforhodamine B (SRB).

[0331] As shown in Figure 2, senescent BV2 cells exposed to masitinib alone ( Figure 2A ) or isoquercetin alone ( Figure 2B ) induced only a slight decrease in cell viability, except for high concentrations of masitinib (2 μM) or isoquercetin (20 μM). In contrast, when senescent BV2 cells were exposed to the combination of masitinib and isoquercetin for 48 h, the number of viable TMZ-induced senescent BV2 cells was significantly reduced ( Figure 2C ). Even at low concentrations of masitinib and isoquercetin (0.1 μM masitinib + 1 μM isoquercetin), the anti-aging effect was evident, with a decrease in senescent cell viability of approximately 30%. Strikingly, the anti-aging effect of the combination of masitinib and isoquercetin was significantly greater than the sum of the anti-aging effects of masitinib alone and isoquercetin alone. For example, the combination of 2 μM masitinib and 20 μM isoquercetin induced a decrease in senescent cell viability of approximately 70% ( Figure 2C ), while 2 μM masitinib alone and 20 μM isoquercetin alone each induced a decrease in senescent cell viability by approximately 25% ( Figure 2A -B).

[0332] In summary, the data obtained with BV2 cells show that masitinib and isoquercetin can each selectively and significantly induce the loss of viability of senescent cells at higher concentrations (e.g., 2 μM masitinib and 20 μM isoquercetin). The data obtained with BV2 cells also show that the combination of masitinib and isoquercetin selectively and significantly induces the loss of viability of senescent cells, even at low concentrations (e.g., 0.1 μM masitinib and 1 μM isoquercetin). Moreover, the data obtained with BV2 cells show that the anti-aging effect of the combination of masitinib and isoquercetin is synergistic, that is, masitinib and isoquercetin work synergistically in selectively and significantly inducing the loss of viability of senescent cells.

[0333] Example 2: Clinical trial investigating the efficacy of a combination of masitinib and isoquercetin for the treatment of COVID-19

[0334] This article describes a randomized, double-blind, placebo-controlled clinical trial designed to evaluate the safety and efficacy of a combination of masitinib and isoquercetin (with best supportive care) in the treatment of hospitalized patients with COVID-19.

[0335] The overall goal of the study in adult patients hospitalized with COVID-19 is to evaluate the efficacy of the combination of masitinib and isoquercetin.

[0336] The study is a randomized, double-blind, placebo-controlled clinical trial in which two different patient groups are defined based on the severity of the disease (defined according to the World Health Organization (WHO) COVID-19 severity criteria), which can also be broadly categorized based on the clinical management of the disease, i.e., not requiring admission to the intensive care unit (ICU) (Group 1) versus requiring ICU admission (Group 2). Each patient group will have a separate control group, thus bringing the total to four treatment groups.

[0337] Overall, 120 patients will be recruited; 60 patients in each group, of whom 30 patients will be randomly assigned in a 1:1 ratio to either the masitinib / isoquercetin (i.e., a combination of masitinib and isoquercetin, with best supportive care) group or the control group (placebo masitinib and placebo isoquercetin, with best supportive care).

[0338] Group 1: Patients who do not require ICU admission (at the time of randomization)

[0339] 30 patients will be randomized to receive masitinib / isoquercetin and best supportive care (excluding hydroxychloroquine and chloroquine). Masitinib will be given orally at a dose of 3 mg / kg / day (mg / kg body weight / day) or 4.5 mg / kg / day. If the safety profile assessed by the Data Safety Monitoring Board (DSMB) is acceptable, patients may receive 3 mg / kg / day of masitinib for at least 4 days, preferably at least 2 days, followed by 4.5 mg / kg / day.

[0340] At least 30 patients with matching baseline characteristics will be included in the control group and will receive placebo masitinib and placebo isoquercetin and best supportive care (excluding hydroxychloroquine and chloroquine).

[0341] Best supportive care was the best available therapy selected by the investigator and included, but was not limited to, oxygenation, analgesics, antithrombotic agents, antiviral agents, and biologic agents.

[0342] Group 2: Patients requiring ICU admission

[0343] 30 patients will be randomized to receive masitinib / isoquercetin and best supportive care (excluding hydroxychloroquine and chloroquine). Masitinib will be given orally at a dose of 3 mg / kg / day or 4.5 mg / kg / day. If the safety profile assessed by the DSMB is acceptable, patients may receive masitinib at 3 mg / kg / day for at least 4 days, preferably at least 2 days, followed by 4.5 mg / kg / day. Patients may or may not receive steroids, according to local procedures.

[0344] At least 30 patients with matching baseline characteristics will be included in the control group and will receive placebo masitinib and placebo isoquercetin and best supportive care (excluding hydroxychloroquine and chloroquine).

[0345] The recommended oral dose of the masitinib / isoquercetin combination is:

[0346] Masitinib: Patients receive a daily dose of 3 mg / kg / day (mg / kg body weight / day) or 4.5 mg / kg / day of masitinib. If the safety profile assessed by the DSMB is acceptable, patients can receive a daily dose of 3 mg / kg / day of masitinib for at least 4 days, preferably at least 2 days, and then receive a daily dose of 4.5 mg / kg / day of masitinib.

[0347] Isoquercetin: The daily dose of isoquercetin is 1 g / day, administered by oral route.

[0348] Masitinib / isoquercetin should be started within 24 hours of cessation of oxygen therapy or discharge from hospital, with a minimum treatment period of 7 days being preferred.

[0349] The study duration was 90 days.

[0350] The WHO COVID-19 severity criteria are as follows:

[0351] - Mild: cases with mild clinical symptoms and no signs of pneumonia in imaging studies;

[0352] -Moderate: Fever and respiratory symptoms with radiologic findings of pneumonia and requiring oxygen (O 2 ): 3L / min <O 2 Cases with blood pressure <5L / min;

[0353] - Severe: Cases that meet any of the following criteria:

[0354] ο Respiratory distress (respiratory rate (RR) ≧ 30 breaths / min);

[0355] ο Resting oxygen saturation in ambient air (SpO 2 )≤93%; or SpO 2 ≤ 97% with O 2>5L / min;

[0356] The ratio of arterial oxygen partial pressure to inspired oxygen concentration (PaO 2 / FiO 2 )≦300mmHg (1mmHg=0.133kPa), PaO in high altitude areas (altitude is above 1000 meters above sea level) 2 / FiO 2 The following formula should be used for correction: PaO 2 / F O 2 [multiplied by] [atmospheric pressure (mmHg) / 760]; and / or

[0357] ο Chest imaging shows significant lesion progression >50% within 24-48 hours;

[0358] - Critical: Patients who meet any of the following criteria:

[0359] oRespiratory failure and requiring mechanical ventilation;

[0360] oShock; and / or

[0361] oOther organ failure requiring ICU care.

[0362] Inclusion criteria:

[0363] 1. Laboratory-confirmed SARS-CoV-2 infection as determined by polymerase chain reaction (PCR) or other commercial or public health assays in any sample ≤72 hours before randomization and / or CT scan following typical radiological findings (abnormal ground-glass opacities and absence of lymphadenopathy, pleural effusion, pulmonary nodules, pulmonary cavitation) 2. Patients hospitalized for treatment of COVID lung disease

[0364] 3. Adult males or females aged ≥18 years at the time of enrollment

[0365] 4. Patients belonging to one of the following two groups:

[0366] - Group 1: patients with moderate and severe lung disease who did not require ICU at initial admission according to WHO COVID-19 severity criteria:

[0367] Moderate cases (score of 4 on the modified WHO 7-point progression scale as described in Table 2 above)

[0368] Cases that meet all of the following criteria:

[0369] · Radiological findings showing fever and respiratory symptoms with pneumonia; and

[0370] Requires 3 L / min to 5 L / min of oxygen to maintain SpO2 >97%.

[0371] or

[0372] Moderate lung disease cases defined by all of the following criteria:

[0373] Requires more than 3 L / min of oxygen;

[0374] Score = 5 on the 10-point WHO Progress Scale; and

[0375] No non-invasive ventilation (NIV) or high-flow oxygen.

[0376] Severe cases (score of 5 on the modified WHO 7-point progression scale as described in Table 2 above)

[0377] Cases that meet any of the following criteria:

[0378] Respiratory distress (RR ≥ 30 breaths / min);

[0379] SpO at rest in ambient air 2 ≤93% ； or SpO 2 ≤ 97% with O 2 >5 L / min; and / or

[0380] ·PaO 2 / FiO 2 ≤300mmHg.

[0381] - Group 2: Patients requiring ICU based on COVID lung disease severity criteria:

[0382] Respiratory failure requiring mechanical ventilation; and

[0383] No do-not-resuscitate order (DNR order).

[0384] 5. Weight > 45 kg and BMI ≥ 18 and ≤ 35 kg / m 2 of patients.

[0385] The primary and secondary endpoints will depend on the patient group tested.

[0386] For group 1 patients (not requiring ICU):

[0387] - Co-primary endpoints

[0388] 1. Survival without the need for a ventilator, including non-invasive ventilation (NIV), on day 14. Therefore, the events considered are the need for a ventilator, including NIV, or death. A new DNR order will be considered the event on the DNR date.

[0389] 2. Early endpoint: clinical status of the patient when the WHO 10-point progression scale score is ≤5 on day 4 (as described in Table 1 above) or when the modified WHO 7-point progression scale is used on day 15 (as described in Table 2 above).

[0390] - Secondary endpoints for Cohort 1 will be as follows:

[0391] · WHO 10-point Progress Scale score at days 4, 7, and 14;

[0392] Overall survival at 14, 28, and 90 days;

[0393] Time to transfer to the ICU;

[0394] · Duration of use of ventilator, NIV or high flow;

[0395] Time of discharge from hospital;

[0396] Time when oxygen is not needed;

[0397] ·Time of negative viral excretion;

[0398] Improvements in biological parameters: estimated glomerular filtration rate (eGFR), C-reactive protein (CRP), myoglobin, creatine phosphokinase (CPK), cardiac troponin, ferritin, lactate, cell blood counts, liver enzymes, lactate dehydrogenase (LDH), D-dimer, albumin, fibrinogen, triglycerides, coagulation tests, urine electrolytes, creatinuria, proteinuria, uricemia, IL6, procalcitonin, immunophenotype, and exploratory tests;

[0399] Clinical status using the following ordinal scale corresponding to the modified WHO 7-point progression scale as described in Table 2 above: 1. Not hospitalized, no limitation of activity; 2. Not hospitalized, limitation of activity; 3. Hospitalized, not requiring supplemental oxygen; 4. Hospitalized, requiring supplemental oxygen; 5. Hospitalized, using noninvasive ventilation or high-flow oxygen device; 6. Hospitalized, receiving invasive mechanical ventilation or ECMO; 7. Death.

[0400] For Group 2 patients (ICU required):

[0401] - Co-primary endpoints

[0402] 1. Cumulative incidence of successful tracheal extubation (defined as extubation duration >48 hours) at day 14. Death or DNR order will be considered as competing events.

[0403] 2. Early endpoint: WHO 10-point Progression Scale score ≤ 7 on day 4 (as described in Table 1 above).

[0404] - Secondary endpoints for Group 2 will be as follows:

[0405] · WHO 10-point Progress Scale score at days 4, 7, and 14;

[0406] Overall survival at 14, 28, and 90 days;

[0407] 28 ventilator-free days;

[0408] ·PaO 2 / FiO 2 The evolution of the ratio;

[0409] Respiratory acidosis on day 4 (arterial pH <7.25, arterial carbon dioxide partial pressure [Paco 2 ]≥60 mmHg, duration>6 hours);

[0410] Time when oxygen is not needed;

[0411] Length of hospital stay;

[0412] ·Time of negative viral excretion;

[0413] ICU discharge time;

[0414] Time of discharge from hospital;

[0415] Improvements in biological parameters (eGFR, CRP, cardiac troponin, urine electrolytes and creatinine, proteinuria, uricemia, IL6, myoglobin, kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), CPK, ferritin, lactate, blood counts, liver enzymes, LDH, D-dimer, albumin, fibrinogen, triglycerides, coagulation tests (including activated partial thromboplastin time), procalcitonin);

[0416] Clinical status using the following ordinal scale corresponding to the modified WHO 7-point progression scale as described in Table 2 above: 1. Not hospitalized, no limitation of activity; 2. Not hospitalized, limited activity; 3. Hospitalized, not requiring supplemental oxygen; 4. Hospitalized, requiring supplemental oxygen; 5. Hospitalized, using noninvasive ventilation or high-flow oxygen device; 6. Hospitalized, receiving invasive mechanical ventilation or ECMO; 7. Death;

[0417] · Rate of renal replacement therapy;

[0418] Ventilation parameters.

[0419] Safety assessment criteria include:

[0420] Number of serious adverse events

[0421] Cumulative incidence of serious adverse events

[0422] Cumulative incidence of grade 3 and 4 adverse events

[0423] Study drug discontinuation (for any reason)

[0424] Example 3: Clinical trial investigating the efficacy of masitinib as a single agent for the treatment of COVID-19

[0425] This article describes a randomized, double-blind, placebo-controlled phase 2 clinical trial designed to evaluate the antiviral efficacy of masitinib in patients with mild to moderate COVID-19 symptoms.

[0426] The overall goal of the study is to evaluate the antiviral efficacy of 3 different doses of masitinib in patients with mild to moderate COVID-19 symptoms.

[0427] In addition, all patients will receive best supportive care and will be randomly assigned to one of the following 3 groups:

[0428] Masitinib 3.0 mg / kg / day for 10 days versus corresponding placebo,

[0429] Masitinib 3.0 mg / kg / day for 2 days, then 4.5 mg / kg / day for 8 days, compared with matching placebo,

[0430] Masitinib 3.0 mg / kg / day for 2 days, then 4.5 mg / kg / day for 2 days, then 6.0 mg / kg / day for 6 days, compared with matching placebo.

[0431] For each group, hydroxychloroquine, chloroquine, and remdesivir were excluded. Best supportive care is the best available therapy selected by the investigator, including but not limited to antipyretics, corticosteroids, oxygenation, analgesics, antithrombotic agents, and antiviral drugs approved for COVID-19. Treatment will be administered for 10 days. Patients will be followed for 1 month.

[0432] 78 patients will be recruited. For each group, 20 patients will be treated with masitinib and 6 patients will be treated with placebo. 50% of the patients will be included in the active and control groups with a score of 2 or 3 on the WHO 10-point Clinical Progression Scale (as described in Table 1 above), and 50% of the patients will be included in the active and control groups with a score of 4 or 5 on the WHO 10-point Clinical Progression Scale (as described in Table 1 above).

[0433] Inclusion criteria:

[0434] 1. Men or non-pregnant women with symptomatic ambulatory mild COVID-19 (scores 2 and 3 on the WHO 10-point Clinical Progression Scale as described in Table 1 above) and

[0435] Age ≥ 75 years

[0436] or 65 years old < age < 74 years old and have the following comorbidities:

[0437] - Complex arterial hypertension

[0438] - Class I obesity: BMI 30 to ≤35 kg / m 2

[0439] -diabetes

[0440] - Obstructive pulmonary disease or respiratory failure

[0441] Or hospitalized adult males or non-pregnant adult females aged ≥18 years with COVID-19 and a score of 4 on the 10-point WHO Clinical Progression Scale as described in Table 1 above with the following comorbidities at the time of enrollment:

[0442] - Complex arterial hypertension

[0443] - Class I obesity: BMI 30 to ≤35 kg / m 2

[0444] -diabetes

[0445] - Obstructive pulmonary disease or respiratory failure

[0446] Or hospitalized adult males or non-pregnant females aged ≥18 years with COVID-19 and a score of 5 on the 10-point WHO Clinical Progression Scale as described in Table 1 above at the time of enrollment.

[0447] 2. According to the investigator's judgment, the patient has symptoms consistent with COVID-19, with onset ≤ 5 days before randomization.

[0448] 3. Positive COVID-19 test ≤72 hours before randomization (as verified by SARS-CoV-2 RT-PCR or other molecular diagnostic assay using appropriate specimens, such as nasal, oropharyngeal, or saliva specimens) and no other explanation for the current clinical condition.

[0449] 4. Weight > 45 kg and BMI ≥ 18 and ≤ 35 kg / m 2 of patients.

[0450] The primary objective was to evaluate the efficacy of masitinib in patients with mild and moderate COVID-19 based on the viral load of the patients after 10 days of treatment. Therefore, the primary endpoint was: change in viral load measured by RT-qPCR in nasal swabs on days 4, 7, and 10.

[0451] Safety assessment criteria include:

[0452] Cumulative incidence of serious adverse events (SAEs)

[0453] Cumulative incidence of grade 3 and 4 adverse events (AEs).

[0454] Discontinuation of investigational medicinal product (for any reason)

[0455] Example 4: In vitro antiviral effect of masitinib

[0456] This example shows that masitinib inhibits both nested viruses and picornaviruses.

[0457] Materials and methods

[0458] Material

[0459] cell

[0460] A549 expressing H2B-mRuby is produced by first infecting A549 cells (ATCCCL-185) with a lentivirus (carrying H2B-mRuby) and FACS sorting mRuby+ cells. They are maintained as a polyclonal population and grown in DMEM+10% BCS (calf serum). These cells are used for all OC43 infections (i.e., HCoV-OC43 infections). Ace2-A549 cells (Blanco-Melo et al., Cell, 181: 1036-1045.e9 (2020), incorporated herein by reference) are used for SARS-CoV-2 infection. They are maintained in DMEM+10% FBS (fetal bovine serum). African green monkey kidney cells (Vero E6) are maintained in DMEM supplemented with 10% FBS, 1% penicillin-streptomycin and 1% HEPES. Huh7 cells were used for picornavirus infection (i.e., infection with one of coxsackievirus B3 (CVB3) or human rhinovirus 2, 14, and 16 (HRV2, 14, 16)). MDCK-SIAT1-TMPRSS2 cells were used for influenza A virus (IAV) infection. A549 cells were maintained in 50:50 DMEM:F-12 medium supplemented with 10% FBS and 1% penicillin-streptomycin for lymphocytic choriomeningitis virus (LCMV) infection.

[0461] Virus

[0462] OC43 (i.e., HCoV-OC43) was obtained from ATCC (VR-1558) and grown and titrated on A549-mRuby cells. SARS-CoV-2 (nCoV / Washington / 1 / 2020) was provided by the National Biocontainment Laboratory (Galveston, TX). VeroE6 cells were used to propagate and titrate SARS-CoV-2. Coxsackievirus B3 or CVB3 (Nancy strain), human rhinovirus (HRV) 2, 14, and 16 were derived from full-length infectious clones and produced in Vero cells (NR-10385, BEI Resources, NIAID, NIH). Recombinant lymphocytic choriomeningitis virus (rLCMV) based on the Armstrong 53b strain was generated as previously described (Flatz et al., Proc. Natl. Acad. Sci. USA, 103:4663-4668 (2006) and Ziegler et al., PLoS Pathog., 12:e1005501 (2016), each of which is incorporated herein by reference). Working stocks were generated in Vero E6 cells, and viral titers were measured using the same cells. The measles virus (MeV) used was derived from a molecular cDNA clone of the Moraten / Schwartz vaccine strain (del Valle et al., J. Virol., 81:10597-10605 (2007), incorporated herein by reference). Recombinant measles virus was engineered to express firefly luciferase ( et al., Viruses, 11:(2019), incorporated herein by reference).

[0463] method

[0464] Drug Screening

[0465] A549-mRuby cells were seeded (3,000 cells per well) in nine 384-well plates using a Multidrop combi. Cells were seeded in a final volume of 30 μL with DMEM + 10% BCS. The next day, 20 μL of OC43 (multiplicity of infection (MOI) 0.3) was added and incubated at 33°C, 5% CO 250 nL (1:1,000 dilution) of an FDA-approved drug library from Selleck (cat# L1300, Selleck) was added. Two columns (32 wells) remained uninfected, and two columns were treated with DMSO and virus (no drug control). Cells were imaged using an IncuCyte S3 to measure cell number on day 0. Cells were incubated at 33°C, 5% CO 2 Incubate for 4 days at room temperature and stain OC43 nuclear protein. All the following steps were performed at room temperature. Cells were fixed in 50 μL 4% PFA / PBS for 15 min, blocked with 50 μL 10% BSA+0.5% Triton X-100 in PBS for 30 min, stained with 50 μL anti-OC43 (cat#MAB9013, Millipore) diluted 1:2,000 in 2% BSA+0.1% Triton X-100 in PBS for 1 hour, washed three times with 50 μL PBS, stained with anti-mouse-AlexaFluor488 diluted 1:1,000 in 2% BSA+0.1% Triton X-100 in PBS for 1 hour, washed 3 times with 50 μL PBS and imaged on IncuCyteS3 (Day 4). Screening was performed twice.

[0466] The following parameters were extracted from the images: cell number at day 0, cell number at day 4, and total OC43 staining intensity at day 4. For analysis, OC43 staining intensity was normalized to the number of cells in the well and further normalized to the mean of the no-drug control, which was set to 100. Compounds that showed a significant effect on cell growth were removed from the analysis. For each plate, a drug was considered a putative hit if it reduced OC43 staining by more than 3 standard deviations relative to the mean of the no-drug control. A drug was considered a hit if it was non-toxic and reduced OC43 staining by more than 3 standard deviations in two replicates. Masitinib was thus identified as a hit with an average % OC43 staining of 12.2 (corresponding to % OC43 staining of 14.4 in screen replicate #1 (normalized to the no drug control in the same plate) and % OC43 staining of 10.1 in screen replicate #2 (normalized to the no drug control in the same plate)), and the number of cells on day 4 divided by the number of cells on day 0 was 4.3 (replicate #1) and 4.7 (replicate #2).

[0467] Dose-response analysis of OC43 and SARS-CoV-2 infection

[0468] Dose response analysis for OC43 infection was similar to that for drug screening, except that cells were plated at 5,000 cells per well and the medium contained 2% BCS instead of 10%. OC43 staining was performed 2 days post infection and its analysis was similar to that described for drug screening. EC50 values ​​were extracted using sigmoid fitting using Matlab.

[0469] All SARS-CoV-2 infections were performed at Howard T. Ricketts Regional Biocontainment Laboratory under biosafety level 3 conditions. Ace2-A549 cells in DMEM + 2% FBS were treated with drugs for 2 hours, with 2-fold dilutions starting from 10 μM, and each assay was performed in triplicate. Cells were infected at an MOI of 0.5 in medium containing the appropriate concentration of drug. After 48 hours, cells were fixed with 3.7% formalin, blocked and probed with a mouse anti-spike protein antibody (GTX632604, GeneTex) diluted 1:1,000 for 4 hours, rinsed and probed with anti-mouse HRP for 1 hour, washed, and then developed with DAB (3,3'-diaminobenzidine) substrate for 10 minutes. Spike protein-positive cells were quantified by light microscopy as blinded samples (n>40).

[0470] For SARS-CoV-2 plaque titers, cell supernatants from the above infections were serially diluted (using 10-fold steps) and used to infect Vero E6 cells for 1 hour. The inoculum was removed and a 1.25% methylcellulose DMEM solution was added to the cells and incubated for 3 days. The plates were fixed in 1:10 formalin for 1 hour, stained with crystal violet for 1 hour and counted to determine plaque forming units (PFU) / ml.

[0471] FlipGFPSARS-CoV-23Clpro assay

[0472] 293T cells were seeded on poly-lysine treated plates 24 hours before transfection. The next day, SARS-CoV-23CLpro plasmid, FlipGFP coronavirus reporter plasmid, Opti-MEM and TransIT-LT (Mirus) were combined and incubated at room temperature for 20 minutes before adding to the cells. At the time of transfection, the indicated concentrations of masitinib were applied to the cells. 24 hours after transfection, cells were fixed with 2% PFA at room temperature for 20 minutes and incubated overnight at 4°C in PBS solution of 1:10,000 Hoescht 33342 (Life Technologies). Quantification was performed by using a CellInsight CX5 (Thermo Scientific) device.

[0473] 3CLpro luciferase reporter assay

[0474] About 16 hours before transfection, 293T cells were seeded in 96-well plates and grown overnight to 70-80% confluence. The next day, cells were transfected with 37.5ng pGlo-30F-VRLQS, 37.5ng SARS-CoV-23CLpro, and 2.5ng pRL-TK (Promega) using Lipofectamine 2000 (Invitrogen) according to the manufacturer's recommendations. After 18 hours, Masitinib (0-10 μM) was added to the cells and incubated for another 6 hours, and then luciferase readout was performed on a Biotek Synergy plate reader as previously described (Kilianski et al., J. Virol., 87: 11955-11962 (2013), incorporated herein by reference). Briefly, 40 μL of growth medium was removed from each well, and then 40 μL of firefly assay buffer (Triton lysis buffer (50 mM Tris, pH 7.0, 75 mM NaCl, 3 mM MgCl) containing 5 mM DTT, 0.2 mM Coenzyme A, 0.15 mM ATP, and 1.4 mg / mL D-luciferin) was added. 2 , 0.25% Triton X-100)) to lyse cells, and provide substrate for firefly luciferase. Read firefly luminescence after 10min, and add 40L Renilla assay buffer (45mM EDTA, 30mM sodium pyrophosphate, 1.4M NaCl, 0.02mM PTC124, 0.003mM coelenterazine h (CTZ-h)) to stop firefly luciferase activity and provide substrate for Renilla luciferase. Read Renilla fluorescence after adding buffer for 2-3min. Firefly luciferase luminescence is normalized to produce normalized luminescence with respect to corresponding Renilla luciferase luminescence.

[0475] 3CLpro kinetic assay

[0476] Cell-free inhibition assays were performed in triplicate at 25°C using a 96-well plate. Reactions containing different concentrations of masitinib (0-100 M) and 3CLpro enzyme (125 nM), 1 mM EDTA, 2 mM DTT in Tris-HCl pH 7.3 were incubated for 20 min. The reaction was then initiated with 5-FAM-TSATLQSGFRK (QXL520) -NH2 probe substrate (1.5 μM). Fluorescence emission intensity was measured (excitation: 490 nm; emission: 520 nm). The data were fitted using a sigmoid curve in Matlab.

[0477] The cloning of 3Clpro (3CL protease) from SARS CoV-2 is based on the original cloning of SARS-CoV 3CLpro (Xue et al., J. Mol. Biol., 366: 965-975 (2007), incorporated herein by reference). The gene encoding 3CLpro from SARS CoV-2 was cloned between the upstream MBP and downstream sequences of GPHHHHHH. Kneller et al. (Kneller et al., Nat. Commun., 11: 3202 (2020), incorporated herein by reference) describes the detailed cloning of pCSGID-Mpro carrying 3CLpro from SARS CoV-2.

[0478] pCSGID-Mpro was transformed into 100 mL of E. coli BL21 (DE3)-Gold (Strategene) under ampicillin (150 mg / L) selection and grown overnight at 37°C. The primer was then transferred to a 4 L LB-Miller culture and grown at 37°C with constant shaking (190 rpm). After reaching an OD600 of ~1, the shaker was set to 4°C. When the temperature reached 18°C, IPTG and K were added, respectively. 2 HPO 4 to 0.2 mM and 40 mM, and the culture was soaked at 18° C. The cells were centrifuged at 4000 g, resuspended in lysis buffer (500 mM NaCl, 5% (v / v) glycerol, 50 mM HEPES pH 8.0, 20 mM imidazole pH 8.0, 1 mM TCEP), and frozen at -80° C.

[0479] Bacterial cells were lysed by sonication and debris was removed by centrifugation at 25,400 x g for 60 min at 4 °C. The clarified supernatant was mixed with 3 mL Ni 2+Sepharose (GE Healthcare Life Sciences) was mixed. The suspension was applied to a Flex-Column (420400-2510) connected to a Vac-Man vacuum manifold. Unbound proteins were washed off using controlled aspiration lysis buffer (160 ml). 3CLpro was eluted using 15 mL of buffer containing 500 mM NaCl, 5% (v / v) glycerol, 50 mM HEPES pH 8.0, 500 mM imidazole pH 8.0, and 1 mM TCEP. Fractions containing 3CLpro were pooled, and rhinovirus 3C His6-tagged protease was added at a protease: protein ratio of 1:25, and incubated overnight at 4°C to cleave the C-terminal His6 tag, thereby generating 3CLpro with authentic N and C termini. The protein solution was concentrated using a 10 kDa MWCO filter (Amicon-Millipore) and subsequently applied to a Superdex 75 column pre-equilibrated with lysis buffer. Fractions containing 3CLpro were pooled together and passed through 2 mL of Ni resin. The flow-through was collected and the lysis buffer was exchanged with crystallization buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 2 mM DTT (1,4-dithiothreitol, Roche, Basel, Switzerland)) using a 10 kDa MWCO filter. The 3CLpro solution was concentrated to 49 mg / mL, aliquoted, frozen and stored at -80°C.

[0480] Crystallization of masitinib and SARS-CoV-23CLpro

[0481] Crystallization was performed using a previous protocol (Kim et al., Methods 55, 12-28 (2011), incorporated herein by reference). 3CLpro was mixed with a DMSO solution of 0.2M masitinib. The final protein concentration was 6.25 mg / mL, and the inhibitor concentration was 8 times higher. The mixture was incubated for 1 hour (at room temperature) and centrifuged at 12,000 x g to remove the precipitate. For crystallization, a sitting drop vapor diffusion method was used in a 96-well CrystalQuick plate (Greiner Bio-One, Monroe, NC, USA) by a Mosquito liquid dispenser (TTP LabTech, Royston, UK), using a protein-to-matrix ratio of 1: 1. Crystallization was performed at 16°C using ProPlex, PACT premier (Molecular dimensions, Cambridge, UK) and TOP96 (Anatrace, Maumee, OH, USA) sieves. The first thin plate crystals (obtained under several conditions one day later) were used as seeds. The best crystals appeared in PACT B7 (0.2M sodium chloride, 0.1MES pH6.0, 20% PEG6000), TOP96 H8 (0.1M ammonium acetate, 0.1M Bis-Tris pH5.5, 17% PEG10000) and Top96 F11 (0.1M Bis-Tris pH6.5; 25% PEG3350). The crystals selected for data collection were treated in their crystallization buffer supplemented with 10-18% glycerol and then rapidly cooled in liquid nitrogen.

[0482] X-ray data collection and structure determination

[0483] Cryogenically cooled crystals (100K) were measured using single wavelength X-ray diffraction experiments at the 19-ID beamline of the Advanced Photon Source at Argonne National Laboratory (Structural Biology Center, Advanced Photon Source at Argonne National Laboratory) (using the SBCcollect program). The intensities of each data set were integrated, scaled, and merged (using the HKL-3000 program suite (Minor et al., Acta Crystallogr. D Biol. Crystallogr., 62: 859-866 (2006), incorporated herein by reference)). The structure of 3CLpro in complex with masitinib was determined using the molecular replacement method (Vagin et al., Acta Crystallogr. D Biol. Crystallogr. 66, 22-25 (2010), incorporated herein by reference)) with the apo form of 3CLpro (PDB code: 7JFQ) as a search template. In the difference Fourier map, additional electron density was observed in the substrate binding site of 3CLpro and was subsequently identified as the contribution of masitinib. The data set was used for further model reconstruction, including building masitinib into additional density using the Coot program (Emsley et al., Acta Crystallogr. D Biol. Crystallogr., 60: 2126-2132 (2004), incorporated herein by reference) and using the program phenix.refine for correction (Terwilliger et al., Acta Crystallogr. D Biol. Crystallogr., 68: 861-870 (2012), incorporated herein by reference) (see Table 3 below). MOLPROBITY (Chen et al., Acta Crystallogr. D Biol. Crystallogr., 66: 12-21 (2010), incorporated herein by reference) was used to verify the stereochemistry of the structure (see Table 3 below).

[0484] The X-ray structure of 3CLpro-bound masitinib has been deposited to PDF under accession number 7JU7.

[0485] Table 3: Antibody binding of masitinib to SARS-CoV-2 3CL protease (also known as M pro )

[0486]

[0487]

[0488] 1 Last resolution bin 2 Molecular replacement method.

[0489] Picornavirus Infection

[0490] Before infection, Huh7 cells were pretreated for two hours. Serum-free DMEM (SFM) was used to dilute the virus to reach an MOI of 0.01. Cell supernatants (collected 24 hours after infection) were diluted in SFM and used to inoculate Vero cells at 37°C for 10-15min. After covering the cells with DMEM containing 2% NBCS and 0.8% agarose, the cells were incubated at 37°C for 2 days. The cells were then fixed with 4% formalin and developed with crystal violet solution (10% crystal violet; Sigma-Aldrich). The number of plaque forming units (PFU / per ml) was then calculated.

[0491] 3C protease activity assay

[0492] Huh7 cells were transfected with LipoD293 (SignaGen Laboratories) and 3C substrate, 3C protease (derived from CVV3) and Renilla transfection control plasmid (siCheck). Proteases and target constructs were generated using previously described protocols (Dial et al., Viruses 11, (2019), incorporated by reference). 24 hours after transfection, cells were combined with firefly substrate (Bright-Glo; Promega) and then with subsequent Renilla (Stop and Glo; Promega) luciferase substrate. The assay was performed using the manufacturer's recommendations (Promega) and the results were quantified using a Veritas microplate luminometer (Turner BioSystems).

[0493] Influenza A infection

[0494] MDCK-SIAT1-TMPRSS2 cells were infected with influenza A / Puerto Rico / 8 / 1934 (PR8) at an MOI of 0.01 TCID50 / cell. After 1 hour of adsorption, the virus was removed and the cells were washed. Masitinib or DMSO was added to the virus growth medium to a final concentration of 10 μM. The supernatant was harvested 20 hours after infection and clarified. The supernatant was titrated using TCID on MDCK-SIAT1-TMPRSS2 cells.

[0495] LCMV infection

[0496] The day before infection, A549 cells were seeded in 12-well culture dishes (80,000 cells per well). Cells were infected with rLCMV at an MOI of 0.01 for one hour at 37°C. The inoculum was removed and the cells were covered with 1 mL of complete medium containing masitinib or DMSO control alone. The supernatant was harvested 48 hours after infection, clarified, and titrated by previously described immunofocus assay (Graham et al., medRxiv2020.07.15.20154443 (2020). doi: 10.1101 / 2020.07.15.20154443 and Ziegler et al., Gen. Virol. 97: 20842089 (2016), both incorporated herein by reference) using mouse anti-LCMV nucleoprotein antibodies (1-1.3) and peroxidase-labeled goat anti-mouse antibodies (SeraCare).

[0497] Measles virus suppression assay

[0498] Vero cells were infected with measles virus expressing luciferase at an MOI of 0.01 for 90 min. The inoculum was removed and fresh medium containing masitinib or DMSO was added to the cells for further culture. Three days later, firefly luciferase activity was measured by adding 0.5 mM D-luciferin to each well and quantified using the Infinite M200 Pro multi-mode microplate reader.

[0499] Statistical analysis

[0500] For all experiments described, sample size (n) refers to independent biological samples tested. All analyses were performed in Matlab. Multiple comparison correction was performed using the FDR method.

[0501] result

[0502] A drug repurposing screen targeting human betacoronavirus OC43 identified masitinib as effective against SARS-CoV-2.

[0503] A library of 1,900 clinically used drugs that were either approved for use in humans or had extensive human safety data (Phase 2 or Phase 3 clinical trials) was screened for their ability to inhibit OC43 infection of the human lung epithelial cell line A549 (expressing the H2BmRuby nuclear reporter). One day after plating, cells were infected at an MOI of 0.3, incubated at 33°C for 1 hour, and then drugs were added to a final concentration of 10 μM. Cells were then incubated at 33°C for 4 days, fixed, and stained for the presence of viral nucleoprotein. Cells were imaged at day 0 (after drug addition) and day 4 (after staining) to determine the effects of drugs on cell growth and OC43 infection.

[0504] The screening was repeated twice and identified 108 drugs, including masitinib, which significantly reduced OC43 infection without significant cytotoxicity. The overall consistency between the two replicates was high (R2 = 0.81). Among the top hits, 29 drugs were reselected for further validation, including masitinib. Thus, masitinib inhibited OC43 infection in a dose-dependent manner, with an EC50 value (drug concentration required to reduce infection by 50%) of 2.1 μM ( Figure 3 ).

[0505] also, Figure 4 The results shown show that masitinib inhibits OC43 replication in primary human airway epithelial cells with an EC50 of 0.58 μM.

[0506] The EC50 value of masitinib against SARS-CoV-2 infection was determined. In a high biocontainment (BSL3) facility, A549 cells overexpressing the angiotensin-converting enzyme 2 (ACE2) receptor were treated with masitinib for 2 hours, infected with SARS-CoV-2 at an MOI of 0.5, incubated for 2 days, fixed, and stained for viral spike protein (as a marker of SARS-CoV-2 infection). After staining, cells were imaged under a microscope to quantify the proportion of infected cells. Masitinib inhibited SARS-CoV-2 infection in a dose-dependent manner with an EC50 value of 3.2 μM ( Figure 5 ).

[0507] The effect of masitinib on viable progeny virus production was evaluated. Cells were treated with 10 μM masitinib for 2 h, infected at an MOI of 0.5, and supernatants were collected 2 days later for titration ( Figure 6 Masitinib completely abolished the production of SARS-CoV-2 progeny (>5-log reduction).

[0508] Therefore, screening identified masitinib as a human-safe drug that can inhibit OC43 and SARS-CoV-2 infection in vitro.

[0509] Masitinib is a true 3CLpro inhibitor

[0510] Masitinib was studied to inhibit the main protease of SARS-CoV-2 (also known as 3CLpro, M pro and nsp5). 3CLpro is essential for the viral replication cycle and is well conserved among coronaviruses. The ability of masitinib to inhibit 3CLpro activity in 293T cells transfected with the FlipGFP reporter system was tested at a single concentration of 10 μM (Anand et al., Science, 300: 1763-1767 (2003), incorporated herein by reference). In this assay, 3CLpro cleavage of the FlipGFP reporter is required to produce GFP fluorescence, and therefore the level of GFP+ cells reports 3CLpro activity. Fig. 7A As shown, masitinib showed a statistically significant decrease in the percentage of GFP expressing cells. Masitinib completely inhibited 3CLpro activity.

[0511] Therefore, the IC50 value (drug concentration resulting in a 50% reduction in enzyme activity) of masitinib inhibition of 3CLpro activity was determined in two different cell-based assays: the same FlipGFP reporter assay as described above ( Figure 7B ), and a luciferase reporter assay adapted for SARS-CoV-224 ( Figure 7C ). These assays determined an IC50 value of 2.5 μM ( Figure 7B -C), similar to that against OC43 infection (2.1 μM, Figure 3 ) and SARS-CoV-2 infection (3.2 μM, Figure 5 ) determined, indicating that masitinib inhibits coronavirus infection by inhibiting 3CLpro activity.

[0512] The IC50 value of masitinib was evaluated in a cell-free assay using purified 3CLpro and a fluorescent substrate, in which the fluorescent signal was released by the proteolytic activity of 3CLpro ( Fig.7D , IC50 = 4.3 μM. See above for a full description of the assay). This assay strongly suggests that a direct interaction of masitinib with the viral protease is responsible for its effect on SARS-CoV-2 replication.

[0513] Finally, in vitro characterization of 3CL inhibition by masitinib was measured in the presence of different substrate concentrations. Fig. 7E Medium. Masitinib is a competitive inhibitor of 3CL activity with a Ki value of 2.58 μM.

[0514] Masitinib inhibits 3CLpro by directly binding to its active site

[0515] To gain further mechanistic understanding of the inhibitory mode of action of masitinib on 3CLpro, the high-resolution structure of 3CLpro bound to masitinib was determined using X-ray crystallography ( Fig. 8A -B). The structure shows that masitinib non-covalently binds to a shallow and elongated groove between domains I and II of 3CLpro and passes through the 3CLpro active site. The enzyme is a dimer and both active sites are occupied by masitinib. Specifically, the pyridine ring of masitinib fits into the S1 substrate pocket of the 3CLpro peptide recognition site. In addition to the hydrophobic and van der Waals interactions between the ring and the pocket-forming residues around it, it also forms a hydrogen bond with His163 located at the bottom of the S1 pocket. The aminothiazole ring of masitinib forms a hydrogen bond with the carbonyl group of His164 and interacts directly with the key catalytic residue -Cys145. The second catalytic residue His41 forms a nearly perfect ππ stack with the hydrophobic toluene ring of masitinib occupying the S2 binding pocket. These three active groups (pyridine, aminothiazole, and toluene ring) contribute to the majority of the interaction between masitinib and 3CLpro, binding key active site residues and effectively blocking peptide substrate access to the protease catalytic dyad, thereby preventing polyprotein cleavage.

[0516] In summary, the results suggest that masitinib, originally designed as a tyrosine-kinase inhibitor and considered for the treatment of a variety of human diseases, possesses potent anti-coronavirus activity through its direct binding and inhibition of the viral main protease.

[0517] Masitinib blocks picornavirus replication by inhibiting picornavirus 3C protease

[0518] Since masitinib directly binds to and inhibits coronavirus 3CL proteases, given the extensive structural homology and substrate specificity shared between these viral proteases, its effectiveness against 3C proteases of picornaviruses (human pathogens that cause a range of diseases, including meningitis, hepatitis, and poliomyelitis) was investigated. Using a luciferase reporter assay (Dial et al., Viruses, 11: (2019), incorporated herein by reference), it was found that masitinib significantly inhibited the activity of 3C proteases in cells ( Fig. 9A Masitinib also effectively blocks the replication of multiple small RNA viruses, namely coxsackievirus B3 (CVB3) and human rhinoviruses 2, 14, and 16 (HRV2, HRV14, HRV16) ( Fig. 9B), but not against other RNA viruses, namely influenza A virus (IAV, Orthomyxoviridae), measles virus (MeV, Paramyxoviridae), lymphocytic choriomeningitis virus (LCMV), and chikungunya virus (CHIKV, Togaviridae) ( Fig.10 ). Thus, masitinib is able to inhibit a variety of coronaviruses and picornaviruses, but not other RNA viruses that do not rely on 3CL-like proteases to complete their life cycle.

[0519] Although masitinib binds to 3CLpro in a non-covalent manner, it shows better in vitro efficacy against SARS-CoV-2 replication than the covalent preclinical 3CLpro inhibitor 13b (Zhang et al., Science, 368: 409-412 (2020)). In addition, although masitinib has a higher EC50 value than the other two preclinical covalent inhibitors 11a and 11b (Dai et al., Science, 368: 1331-1335 (2020)), it shows better inhibition of progeny production at 10 μM (masitinib exceeds 5-log, compared to 2-log for 11a and 11b).

[0520] Example 5: In vivo inhibitory effect of masitinib on SARS-CoV-2

[0521] Materials and methods

[0522] Seven-week-old female Tg(K18-hACE2)2Prlmn (Jackson Laboratories, Bar Harbor, ME) mice were intranasally delivered with 2×10 4 pfu of USA-WA1 / 2020SARS-CoV-2 (2019-nCoV) for virus challenge. Mock-infected female mice received 50 μL of PBS instead of virus challenge. Mice were treated by intraperitoneal injection of 100 μL of PBS or masitinib at a concentration ranging from 25 mg / kg to 50 mg / kg twice a day (i.e., bid) starting 12 hours after inoculation. The clinical symptoms and weight loss of mice were tracked twice a day for 6 days after the challenge. Categories included in the clinical score included the appearance of posture and fur (erection of hair) (0-3 points, the lower the score, the better the posture and appearance), and the development of respiratory distress (0-3 points, the lower the score, the lighter the respiratory distress). On the fourth and sixth days after the challenge, five mice in each treatment group were killed and the lungs and nasal conchae were collected to evaluate the viral load. All mouse work was approved by the Animal Care and Use Committee, and all procedures were performed in a certified animal biosafety level 3 laboratory.

[0523] result

[0524] The SARS-CoV-2 viral load in mice was measured 4 and 6 days after infection with SARS-CoV-2. Mice were treated with masitinib (25 or 50 mg / kg, bid, ip) or PBS. Fig.11A -B, masitinib induced a significant reduction in SARS-CoV-2 viral load in the lungs and nasal turbinates.

[0525] Clinical scores of mice were measured 1-6 days after infection with SARS-CoV-2. Mice were treated with masitinib (25 or 50 mg / kg, bid, ip) or PBS. Fig.12 As shown, masitinib (at a dose of 25 mg / kg, bid, ip or a dose of 50 mg / kg, bid, ip) induced a significant decrease in clinical scores, that is, significantly improved the clinical status of mice.

[0526] Thus, the results presented here demonstrate the potent in vivo therapeutic effect of masitinib in treating SARS-CoV-2 infection.

Claims

1. Use of masitinib or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causing COVID-19 in a subject in need thereof.

2. The use according to claim 1, wherein the masitinib or a pharmaceutically acceptable salt thereof is used for combined administration with isoquercetin.

3. The use according to claim 2, wherein the masitinib or a pharmaceutically acceptable salt thereof is used for combined administration with isoquercetin at a dose ranging from 0.4 g / day to 2 g / day.

4. The use according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt of masitinib is masitinib mesylate.

5. The use according to any one of claims 1 to 3, wherein the masitinib or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 1 mg / kg / day to 12 mg / kg / day (mg / kilogram body weight / day). 6 . The use according to claim 5 , wherein the masitinib or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 3 mg / kg / day to 6 mg / kg / day.

7. The use according to any one of claims 1 to 3, wherein the masitinib or a pharmaceutically acceptable salt thereof is administered at an initial dose of about 3 mg / kg / day for a period of at least 2 days and thereafter at a dose of about 4.5 mg / kg / day, with toxicity management at each dose increase.

8. The use according to any one of claims 1 to 3, wherein the subject exhibits at least one risk factor that may lead to an increased risk of developing COVID-19.

9. The use according to any one of claims 1 to 3, wherein the subject has mild to moderate COVID-19.

10. The use according to claim 9, wherein the subject has moderate COVID-19.

11. The use according to any one of claims 1 to 3, wherein the subject suffers from severe COVID-19.

12. The use according to any one of claims 1 to 3, wherein the subject suffers from critical COVID-19.

13. The method of any one of claims 1 to 3, wherein the subject suffers from COVID-19 and scores in the range of 2 to 9 on the World Health Organization (WHO) COVID-19 10-point progression scale.

14. The method of claim 13, wherein the subject has COVID-19 and scores 2 or 3 on the WHO COVID-19 10-point progression scale.

15. The use according to claim 13, wherein the subject has COVID-19 and scores in the range of 4 to 6 on the WHO COVID-19 10-point progression scale.

16. The method of claim 13, wherein the subject has COVID-19 and scores 4 or 5 on the WHO COVID-19 10-point progression scale.

17. The use according to any one of claims 1 to 3, wherein the subject has COVID-19 and scores in the range of 2 to 6 on the modified WHO COVID-19 7-point progression scale.

18. The use according to claim 17, wherein the subject has COVID-19 and scores in the range of 2 to 5 on the modified WHO COVID-19 7-point progression scale.

19. The method of claim 17, wherein the subject has COVID-19 and scores 4 or 5 on the modified WHO COVID-19 7-point progression scale.

20. The use according to any one of claims 1 to 3, wherein the masitinib or a pharmaceutically acceptable salt thereof is for administration together with at least one additional pharmaceutically active agent.

21. The use according to claim 20, wherein the at least one additional pharmaceutically active agent is selected from antiviral agents, anti-interleukin 6 (anti-IL6) agents, protease inhibitors, Janus-associated kinase (JAK) inhibitors, BXT-25, brilacidin, dehydroandrographolide succinate half ester, APN01, fingolimod, methylprednisolone, thalidomide, bevacizumab, sildenafil citrate, interferon, carlimycin, and any mixture thereof.

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