WO2023222332A1 - Diphenyl ureas for the treatment of viral infections - Google Patents

Diphenyl ureas for the treatment of viral infections Download PDF

Info

Publication number
WO2023222332A1
WO2023222332A1 PCT/EP2023/060613 EP2023060613W WO2023222332A1 WO 2023222332 A1 WO2023222332 A1 WO 2023222332A1 EP 2023060613 W EP2023060613 W EP 2023060613W WO 2023222332 A1 WO2023222332 A1 WO 2023222332A1
Authority
WO
WIPO (PCT)
Prior art keywords
compounds
sars
infection
cov
subject
Prior art date
Application number
PCT/EP2023/060613
Other languages
French (fr)
Inventor
Ulrich Betz
Sven Lindemann
Original Assignee
Merck Patent Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Merck Patent Gmbh filed Critical Merck Patent Gmbh
Publication of WO2023222332A1 publication Critical patent/WO2023222332A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4184—1,3-Diazoles condensed with carbocyclic rings, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • Diphenyl urea is an important link structure in the design of active substance for treating cancer due to its near-perfect binding with certain acceptors and has demonstrated many activities against several human cancer cell lines. Diphenyl urea is utilized to treat cancer by inhibiting cell signaling transduction, such as RAS-RAFMEK- ERK signaling pathway and PI3K-Akt-mTOR pathway.
  • cell signaling transduction such as RAS-RAFMEK- ERK signaling pathway and PI3K-Akt-mTOR pathway.
  • this structure inhibits tumor cell growth by inhibiting receptor tyrosine kinases multiply, such as Vascular Endothelial Growth Factor Receptors (VEGFRs), Platelet-Derived Growth Factor Receptors (PDGFRs), Epidermal Growth Factor Receptors (EGFRs) (Yi-Cong et al., Mini-Reviews in Organic Chemistry, 16,7, 2019).
  • VEGFRs Vascular Endothelial Growth Factor Receptors
  • PDGFRs Platelet-Derived Growth Factor Receptors
  • EGFRs Epidermal Growth Factor Receptors
  • Coronaviruses are positive-sense, single-stranded RNA (ssRNA) viruses of the order Nidovirales, in the family Coronaviridae .
  • ssRNA single-stranded RNA
  • SARS-CoV-1 severe acute respiratory syndrome
  • MERS-CoV Middle East respiratory syndrome
  • SARS-CoV-2 COVID-19
  • SUBSTITUTE SHEET (RULE 26) the causative agent of SARS epidemic of 2002-03 (Fung, et al, Annu. Rev. Microbiol. 2019. 73:529-57). Severe disease has been reported in approximately 15% of patients infected with SARS-CoV-2, of which one third progress to critical disease (e.g. respiratory failure, shock, or multi organ dysfunction (Siddiqi, et al, J. Heart and Lung Trans. (2020), doi: https://doi.Org/10.1016/j.healun.2020.03.012, Zhou, et al, Lancet 2020; 395: 1054-62.
  • critical disease e.g. respiratory failure, shock, or multi organ dysfunction
  • the virus has a high transmission rate, likely linked to high early viral loads and lack of pre-existing immunity (He, et. al, Nat Med 2020 https://doi.org/10.1038/s41591-020-0869-5). It causes severe disease especially in the elderly and in individuals with comorbidities.
  • the global burden of COVID- 19 is immense, and therapeutic approaches are increasingly necessary to tackle the disease.
  • Intuitive anti-viral approaches including those developed for enveloped RNA viruses like HIV- 1 (lopinavir plus ritonavir) and Ebola virus (remdesivir) have been implemented in testing as investigational drugs (Grein et al, NEJM 2020 https://doi.org/10.1056/NEJMoa2007016; Cao,et al, NEJM 2020 DOI: 10.1056/NEJMoa2001282). But given that many patients with severe disease present with immunopathology, host-directed immunomodulatory approaches are also being considered, either in a staged approach or concomitantly with antivirals (Metha, et al, The Lancet 2020; 395(10229) DOI: https://doi.org/10.
  • Molnupiravir (MK-4482/EIDD-2801) is an investigational, orally administered form of a potent ribonucleoside analog that inhibits the replication of SARS-CoV-2, the causative agent of COVID-19. Molnupiravir has been shown to be active in several preclinical models of SARS- CoV-2, including for prophylaxis, treatment, and prevention of transmission (DOI: 10.1101/2021.06.17.21258639).
  • Nafamostat a serine protease inhibitor
  • COVID-19 pneumonia https://doi.org/10.1128/AAC.00754-20.
  • Diphenyl urea inhibitbitors with antiviral activity against RNA viruses
  • kinases represent an attractive target for broad-spectrum therapy [1] in general but it is unclear what is the best combination of kinases to be inhibited.
  • Garcia et al [doi: https://doi.org/10.1101/2020.06.24.150326] mention mTOR-P 13K-AKT, ABL-25 BCR/MAPK and DNA Damage Response.
  • JAK inhibitors JAK inhibitors have shown significant benefit and have been used in treating several autoimmune diseases and inflammatory diseases, like rheumatoid arthritis and inflammatory bowl disease.
  • JAK-inhibitors showed favorable outcome in Covid- 19, in terms of recovery rate, shortened time to recovery, reduction risk of clinical deterioration, and reduction of mortality rate [Limen et al., Expert Review of Anti-infective Therapy https://doi.org/10.1080/14787210.2021.1982695].
  • Another publication analyzed reports on safety and efficacy of JAK-inhibitors and found that they decreased the use of invasive mechanical ventilation and increased survival [Chen et al., Leukemia 2021, 35:2616-2620, https://doi.org/10.1038/s41375-021-01266-6].
  • VEGF is deemed a promising therapeutic target in suppressing inflammation during SARS-CoV2 infection with neurological symptoms
  • Zambrana et al. [2021, Scientific Reports 11: 18985, https://doi.org/10.1038/s41598-021-98289-x] found that NO and VEGF signaling are potential molecular pathways whose function are very similar with several observed COVID-19 symptoms.
  • VEGF-signaling we suggest as a putative target gene KDR (VEGFR-2).
  • SUBSTITUTE SHEET (RULE 26) demonstrated a higher percentage of survival in comparison to vehicle and a significant reduced virus load.
  • the invention provides compounds 1 -7 for use in the treatment of viral infections in a subject in need thereof.
  • the compounds are subject matter of W02006040056.
  • the viral infection is a single-strand RNA viral infection.
  • the viral infection is a coronavirus infection.
  • the viral infection is a SARS-CoVl, MERS- CoV, or SARS-CoV-2 infection.
  • the viral infection is a SARS-CoV-2 infection.
  • a second embodiment is a method of treating a coronavirus infection in a subject in need thereof, comprising administering an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7, or a pharmaceutically acceptable salt thereof, to the subject.
  • the administration of a diphenyl urea selected but not limited from the group of compounds 1 -7 reduces the viral load in the subject.
  • the subject has a mild to moderate SARS-CoV-2 infection.
  • the subject is asymptomatic at the start of the administration regimen.
  • R2 H, -CO-NHR, -NH-CO-OR with
  • R alkyl
  • SARS-CoV-2 directly enters cells expressing ACE2 via receptor-mediated endocytosis (Hoffmann, et al). Successful viral replication requires host endosome acidification
  • SUBSTITUTE SHEET (RULE 26) to release the viral genome into the host cytosol.
  • Innate immune cells like monocytes, macrophages and neutrophils do not highly express ACE2, but have abundant Fc receptors (Zou, et al; Qi, et al; Lu, et al, Nat. Rev. Imm. 2018 https://doi.org/10.1038/nri.2017.106).
  • stage II Fig.l
  • antibodies that bind the virus can mediate viral uptake into myeloid cell endosomes via Fc receptors (FcR) or complement receptors (CR) (Lu, et al; Dandekar, et al, Nat. Rev. Imm.
  • ACE2 FcR and CR present three mechanisms how SARS-CoV-2 can enter endosomes and trigger hyperinflammation leading to cytokine storm and severe disease.
  • ssRNA virus can induce NETosis in neutrophils (Saitoh, et al, Cell Host Microbe (2012), 19;12(1): 109-16) leading to release of DNA and RNA, creating a feed-forward loop to further fuel inflammation (Herster et al, Nat Commun 2020; 11, 105 https://doi.org/10.1038/s41467-019-13756-4), which has been proposed as a driver of severe COVID-19 (Barnes, et al, I Exp med 2020; 217 (6) https://doi.org/10.1084/jem.20200652).
  • SARS-CoV-1 derived ssRNA has been shown to mediate severe lung pathology in animal models and presents as a potential driver of virus- associated cytokine storm (Li, et al, Microbes Infect 2013; 15 (2) 88-95. https://doi.Org/10.1016/j.micinf.2012.10.008). Being able to slow the viral reproduction in the early stages of infection may allow the subject to avoid severe disease.
  • COVID-19 is the name of the disease which is caused by a SARS-CoV-2 infection. While care was taken to describe both the infection and disease with accurate terminology, “COVID-19” and “SARS-CoV-2 infection” are meant to be roughly equivalent terms.
  • “mild to moderate” COVID-19 occurs when the subject presents as asymptomatic or with less severe clinical symptoms (e.g., low grade or no fever ( ⁇ 39.1°C), cough, mild to moderate discomfort) with no evidence of pneumonia, and generally does not require medical attention.
  • “moderate to severe” infection generally patients present with more severe clinical symptoms (e.g., fever >39.1°C, shortness of breath, persistent cough, pneumonia, etc.).
  • “moderate to severe” infection typically requires
  • SUBSTITUTE SHEET (RULE 26) medical intervention, including hospitalization.
  • a subject can transition from “mild to moderate” to “moderate to severe” and back again in one course of bout of infection.
  • Treatment of CO VID-19 using the methods of this invention include administration of an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention at any stage of the infection to prevent or reduce the symptoms associated therewith.
  • subjects will be administered an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention after definitive diagnosis and presentation with symptoms consistent with a SARS-CoV2 infection, and administration will reduce the severity of the infection and/or prevent progression of the infection to a more severe state.
  • the clinical benefit upon such administration is described in more detail in the sections below.
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (Ci-4alkyl)4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.
  • structures depicted herein are also meant to include all isomeric (e g , enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention.
  • structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are
  • the group comprises one or more deuterium atoms.
  • the term “patient” or “subject”, as used herein, means an animal, preferably a human. However, “subject” can include companion animals such as dogs and cats.
  • the subject is an adult human patient.
  • the subject is a pediatric patient.
  • Pediatric patients include any human which is under the age of 18 at the start of treatment.
  • Adult patients include any human which is age 18 and above at the start of treatment.
  • the subject is a member of a high-risk group, such as being over 65 years of age, immunocompromised humans of any age, humans with chronic lung conditions (such as, asthma, COPD, cystic fibrosis, etc.), and humans with other co-morbidities.
  • the other co-morbidity is obesity, diabetes, and/or hypertension.
  • compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir.
  • the compositions are administered orally.
  • the oral formulation of a compound of the invention is administered orally.
  • the oral formulation is a solution or suspension which may be given to a subject in need thereof via mouth or nasogastric tube. Any oral formulations of the invention may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.
  • compositions of this invention are orally administered in any orally acceptable dosage form.
  • exemplary oral dosage forms are capsules, tablets, aqueous suspensions or solutions.
  • carriers commonly used include lactose and corn starch.
  • Lubricating agents, such as magnesium stearate, are also used.
  • SUBSTITUTE SHEET typically added.
  • useful diluents include lactose and dried corn starch.
  • the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents are optionally also added.
  • compositions of the present invention that are optionally combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration.
  • provided compositions should be formulated so that a dosage of between 0.1 - 100 mg/kg body weight/day of the compound can be administered to a patient receiving these compositions.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention act as host-targeting antivirals by blocking the host pyrimidine synthesis, and thereby blocking the viral replication.
  • the total amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 administered to the subject in need thereof is generally in the range from 0.2 to 90 mg/kg of body weight of the recipient (mammal) per day and particularly typically in the range from 1 to 10 mg/kg of body weight per day.
  • the actual amount per day for an adult mammal weighing 70 kg is usually between 70 and 700 mg, where this amount can be administered as an individual dose per day or usually in a series of part-doses (such as, for example, two, three, four, five or six) per day, so that the total daily dose is the same.
  • An effective amount of a salt or solvate or of a physiologically functional derivative of the diphenyl urea selected but not limited from the group of compounds 1 -7 can be determined as the fraction of the effective amount of the compound per se.
  • diphenyl urea selected but not limited from the group of compounds 1 -7 are administered for a period of about 7 days to about 28 days. In one aspect of any of the above embodiments, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered for about 14 days.
  • the subject is suffering from COVID-19 pneumonia. In one embodiment of this invention, the subject is suffering from one or more of
  • SUBSTITUTE SHEET (RULE 26) symptoms selected from chest congestion, cough, blood oxygen saturation (SpCh) levels below 94%, shortness of breath, difficulty breathing, fever, chills, repeated shaking with chills, muscle pain and/or weakness, headache, sore throat and/or new loss of taste or smell.
  • the subject is being treated inpatient in a hospital setting. In another embodiment, the subject is being treated in an outpatient setting. In one aspect of the preceding embodiments, the subject may continue administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are after being transitioned from being treated from an inpatient hospital setting to an outpatient setting.
  • the administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are results in one or more clinical benefit.
  • the one or more clinical benefit is selected from the group comprising: reduction of duration of a hospital stay, reduction of the duration of time in the Intensive Care Unit (ICU), reduction in the likelihood of the subject being admitted to an ICU, reduction in the rate of mortality, reduction in the likelihood of kidney failure requiring dialysis, reduction in the likelihood of being put on non-invasive or invasive mechanical ventilation, reduction of the time to recovery, reduction in the likelihood supplemental oxygen will be needed, improvement or normalization in the peripheral capillary oxygen saturation (SpCh levels) without mechanical intervention, reduction of severity of the pneumonia as determined by chest imaging (eg, CT or chest X ray), reduction in the cytokine production, reduction of the severity of acute respiratory distress syndrome (ARDS), reduction in the likelihood of developing ARDS, clinical resolution of the COVID-19 pneumonia, improvement of the PaO2/FiO2 ratio, and
  • the one or more clinical benefits includes the improvement or normalization in the peripheral capillary oxygen saturation (SpO2 levels) in the subject without mechanical ventilation or extracorporeal membrane oxygenation.
  • SpO2 levels peripheral capillary oxygen saturation
  • the one of more clinical benefits is reduction in the likelihood of being hospitalized, reduction in the likelihood of ICU admission, reduction in the likelihood being intubated (invasive mechanical ventilation), reduction in the likelihood supplemental oxygen will be needed, reduction in the length of hospital stay, reduction in the
  • SUBSTITUTE SHEET (RULE 26) likelihood of mortality, and/or a reduction in likelihood of relapse, including the likelihood of rehospitalization.
  • the invention also provides a method of treating a viral infection caused by F Herpes simplex virus (HSV), human cytomegalo virus (HCMV), Dengue virus, zoonotic pandemic viruses and infections by coronaviruses in a subject in need thereof comprising administering an effective amount of a compound of the invention to the subject.
  • An amount effective to treat or inhibit a viral infection is an amount that will cause a reduction in one or more of the manifestations of viral infection, such as viral lesions, viral load, rate of virus production, and mortality as compared to untreated control subjects.
  • One embodiment of the invention is a method of treating a coronavirus infection in a subject in need thereof, comprising administering an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 are, or a pharmaceutically acceptable salt thereof, to the subject.
  • the subject is infected with Severe acute respiratory -related coronavirus (SARS-CoV-1, SARS-CoV-2) and Middle East Syndrome coronavirus (MERS).
  • SARS-CoV-1, SARS-CoV-2) Middle East Syndrome coronavirus
  • MERS Middle East Syndrome coronavirus
  • the administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are results in the reduction of the viral load in the subject.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to COVID-19 pneumonia developing. In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to the subject developing a cytokine storm Tn another embodiment, the subject has a mild to moderate SARS-CoV-2 infection. In a further embodiment, the subject is asymptomatic at the start of the administration regimen. In another embodiment, the subject has had known contact with a patient who has been diagnosed with a SARS-CoV-2 infection. In an additional embodiment, the subject begins administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are prior to being formally diagnosed with COVID- 19.
  • the one or more clinical benefits is shortening the duration of infection, reduction of the likelihood of hospitalization, reduction in the likelihood
  • the one or more clinical benefits is avoidance of a significant proinflammatory response.
  • the one or more clinical benefit is the failure of the subject to develop significant symptoms of COVTD-19.
  • the compounds of the invention can be administered before or following an onset of SARS-CoV-2 infection, or after acute infection has been diagnosed in a subject.
  • a therapeutically relevant effect relieves to some extent one or more symptoms of a disorder, or returns to normality, either partially or completely, one or more physiological or biochemical parameters associated with or causative of a disease or pathological condition.
  • Monitoring is considered as a kind of treatment provided that the compounds are administered in distinct intervals, e.g. in order to boost the response and eradicate the pathogens and/or symptoms of the disease.
  • the methods of the invention can also be used to reduce the likelihood of developing a disorder or even prevent the initiation of disorders associated with COVID-19 in advance of the manifestation of mild to moderate disease, or to treat the arising and continuing symptoms of an acute infection.
  • Treatment of mild to moderate CO VID-19 is typically done in an outpatient setting.
  • Treatment of moderate to severe COVID-19 is typically done inpatient in a hospital setting. Additionally, treatment can continue in an outpatient setting after a subject has been discharged from the hospital.
  • the invention furthermore relates to a medicament comprising at least one diphenyl urea selected but not limited from the group of compounds 1 -7 are according to the invention, or a pharmaceutically salts thereof.
  • a “medicament” in the meaning of the invention is any agent in the field of medicine, which comprises one or more compounds of the invention or preparations thereof (e.g. a pharmaceutical composition or pharmaceutical formulation) and can be used in prophylaxis, therapy, follow-up or aftercare of patients who suffer from clinical symptoms and/or known exposure to COVID- 19.
  • the active ingredient may be administered alone or in combination with one or more additional therapeutic agents.
  • a synergistic or augmented effect may be achieved by using more than one compound in the pharmaceutical composition.
  • the active ingredients can be used either simultaneously or sequentially.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more additional therapeutic agents.
  • the one or more additional therapeutic agents is selected from anti-inflammatories, antibiotics, anti-coagulants, antiparasitic agent, antiplatelet agents and dual antiplatelet therapy, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, beta-blockers, statins and other combination cholesterol lowering agents, specific cytokine inhibitors, complement inhibitors, anti-VEGF treatments, JAK inhibitors, BTK inhibitors, TLR(7/8) inhibitors, immunomodulators, anti-inflammasone therapies, sphingosine-1 phosphate receptors binders, N-methyl-d-aspartate (NDMA) receptor glutamate receptor antagonists, corticosteroids, Granulocyte-macrophage colony-stimulating factor (GM-CSF), anti-GM-CSF, interfer
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with another antiviral agent.
  • the antiviral agent is remdesivir.
  • the antiviral agent is lopinavir-ritonavir, alone or in combination with ribavirin and interferon-beta.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administrated in combination with a broad-spectrum antibiotic.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 is administered in combination with chloroquine or hydroxychloroquine. In one aspect of this embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 is is further combined with azithromycin.
  • the diphenyl urea selected but not limited from the group of compound 1 -7 is is administered in combination with one or more additional therapeutic agents selected from hydroxychloroquine, chloroquine, ivermectin, tranexamic acid, nafamostat, virazole, ribavirin, lopinavir/ritonavir, favipiravir, arbidol, leronlimab, interferon beta- la, interferon beta- lb, beta-interferon, azithromycin, nitr zoxamide, lovastatin, clazakizumab, adalimumab, etanercept, golimumab, infliximab, sarilumab, tocilizumab, anakinra, emapalumab, pirfenidone, belimumab, rit
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more anti-inflammatory agent.
  • the anti-inflammatory agent is selected from corticosteroids, steroids, COX-2 inhibitors, and non-steroidal anti-inflammatory drugs (NSAID).
  • the anti-inflammatory agent is diclofenac, etodolac, fenoprofen, flurbirprofen, ibuprofen, indomethacin, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, sulindac, tolmetin, celecoxib, prednisone, hydrocortisone, fludocortisone, bethamethasone, prednisolone, triamcinolone, methylprednisone, dexamethasone, fluticasone, and budesonide (alone or in combination with formoterol, salmeterol, or vilanterol).
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more immune modulators.
  • the immune modulator is a calcineurin inhibitor, antimetabolite, or alkylating agent.
  • the immune modulator is selected from azathioprine, mycophenolate mofetil, methotrexate, dapson, cyclosporine, cyclophosphamide, and the like.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antibiotics.
  • the antibiotic is a broad-spectrum antibiotic.
  • the antibiotic is a penicillin, anti-staphylococcal penicillin, cephalosporin, aminopenicillin (commonly administered with a betalactamase inhibitor), monobactam, quinoline, aminoglycoside, lincosamide, macrolide, tetracycline, glycopeptide, antimetabolite or nitroimidazole.
  • the antibiotic is selected from penicillin G, oxacillin, amoxicillin, cefazolin, cephalexin, cephotetan, cefoxitin, ceftriazone, augmentin, amoxicillin, ampicillin (plus sulbactam), piperacillin (plus tazobactam), ertapenem, ciprofloxacin, imipenem, meropenem, levofloxacin, moxifloxacin, amikacin, clindamycin, azithromycin, doxycycline, vancomycin, Bactrim, and metronidazole.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more anti-coagulants.
  • the anti-coagulant is selected from apixaban, dabigatran, edoxaban, heparin, rivaroxaban, and warfarin.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antiplatlet agents and/or dual antiplatelet therapy.
  • the antiplatelet agent and/or dual antiplatelet therapy is selected from aspirin, clopidogrel, dipyridamole, prasugrel, and ticagrelor.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more ACE inhibitors.
  • the ACE inhibitor is selected from benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril and trandolapril.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more angiotensin II receptor blockers.
  • the angiotensin II receptor blocker is selected from azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more beta-blockers.
  • the beta-blocker is selected from acebutolol, atenolol, betaxolol, bisoprolol/hydrochlorothiazide, bisoprolol, metoprolol, nadolol, propranolol, and sotalol.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more alpha and beta-blocker.
  • the alpha and beta-blocker is carvedilol or labetalol hydrochloride.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more interferons, such as interferon-beta.
  • interferon-beta is interferon-beta-1.
  • the interferon-beta is interferon-beta- la.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more angiotensin receptor- neprilysin inhibitors.
  • the angiotensin receptor-neprilysin inhibitor is sacubitril/valsartan.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more calcium channel blockers.
  • the calcium channel blocker is selected from amlodipine, diltiazem, felodipine, nifedipine, nimodipine, nisoldipine, and verapamil.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more vasodilators.
  • the one or more vasodilator is selected from isosorbide dinitrate, isosorbide mononitrate, nitroglycerin, and minoxidil.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more diuretics.
  • the one or more diuretics is selected from acetazolamide, amiloride,
  • SUBSTITUTE SHEET (RULE 26) bumetanide, chlorothiazide, chlorthalidone, furosemide, hydrochlorothiazide, indapamide, metalozone, spironolactone, and torsemide.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more muscle relaxants.
  • the muscle relaxant is an antispasmodic or antispastic.
  • the one or more muscle relaxants is selected from casisoprodol, chlorzoxazone, cyclobenzaprine, metaxalone, methocarbamol, orphenadrine, tizanidine, baclofen, dantrolene, and diazepam.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antiviral medications.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more additional therapeutic agents selected from antiparasitic drugs (including, but not limited to, hydroxychloroquine, chloroquine, ivermectin), antivirals (including, but not limited to, tranexamic acid, nafamostat, virazole [ribavirin], lopinavir/ritonavir, favipiravir, leronlimab, interferon beta-la, interferon beta-lb, beta-interferon), antibiotics with intracellular activities (including, but not limited to azithromycin, nitrazoxamide), statins and other combination cholesterol lowering and antiinflammatory drugs (including, but not limited to, lovastatin), specific cytokine inhibitors (including, but not limited to, clazakizumab, adalimumab, etanercept, golimumab
  • SUBSTITUTE SHEET (RULE 26) mesylate, arbidol, brilacidin, IFX-1, PAX-1-001, BXT-25, NP-120, intravenous immunoglobulin (IVIG), and solnatide.
  • the diphenyl urea selected but not limited from the group of compounds 1 -7 with one or more additional therapeutic agents reduces the effective amount (including, but not limited to, dosage volume, dosage concentration, and/or total drug dose administered) of diphenyl urea selected but not limited from the group of compounds 1 -7 and/or the one or more additional therapeutic agents administered to achieve the same result as compared to the effective amount administered when the diphenyl urea selected but not limited from the group of compounds 1 -7 or the additional therapeutic agent is administered alone.
  • the combination of a diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent reduces the total duration of treatment compared to administration of the additional therapeutic agent alone.
  • the combination of a diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent reduces the side effects associated with administration of the additional therapeutic agent alone. In some embodiments, the combination of an effective amount of the diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent is more efficacious compared to an effective amount of the diphenyl urea selected but not limited from the group of compounds 1 -7 or the additional therapeutic agent alone. In one embodiment, the combination of an effective amount of the dual diphenyl urea selected but not limited from the group of compounds 1 -7 with the one or more additional therapeutic agent results in one or more additional clinical benefits than administration of either agent alone.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a viral infection, or one or more symptoms thereof, as described herein.
  • treatment is administered after one or more symptoms have developed.
  • treatment is administered in the absence of symptoms.
  • treatment is administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a known exposure to an infected person and/or in light of comorbidities which are predictors for severe disease, or other susceptibility factors).
  • Compound 1 -7 can be synthesized as described W02006040056, page 189, Synthesis of ureas.
  • Example 2 SARS-CoV-2/Calu-3 high-content screening assay ( Figure 14 and Figure 15)
  • Compounds are acoustically transferred into 384-well pci ear-bottom plates (Greiner, Part. No. 781090-2B) before seeding Calu-3 cells in assay media (MEM with 2% FBS) at a density of 5,000 cells per 20 pL per well.
  • the plated cells are transported to the BSL3 facility where SARS-CoV-2 (strain USA-WA1/2020 propagated in Vero E6 cells) diluted in assay media is added at an MOI between 0.75 and 1 to achieve ⁇ 30 - 60% infected cells. Plates are incubated for 48 h at 34°C 5% CO2, and then fixed with a final concentration of 4% formaldehyde.
  • Plates are imaged using the ImageXpress Micro Confocal High-Content Imaging System (Molecular Devices) with a 10x objective, with 4 fields imaged per well. Images are analyzed using the Multi -Wavelength Cell Scoring Application Module (MetaXpress), with DAPI staining identifying the host-cell nuclei (the total number of cells in the images) and the SARS-CoV-2 immunofluorescence signal leading to identification of infected cells.
  • ImageXpress Multi -Wavelength Cell Scoring Application Module
  • Curves are fitted as either increasing or decreasing and noted as such in the data output. This is of particular note for the cell count readout from the SARS-CoV-2 infection assay which captures both an antiviral effect, protection from virus-induced cell death (increasing), and cellular toxicity (decreasing).
  • Compounds are acoustically transferred into 384-well pclear-bottom plates (Greiner, Part. No. 781090-2B) and HeLa-ACE2 cells are seeded in the plates in 2% FBS at a density of 1.0x 103 cells per well. Plated cells are transported to the BSL3 facility where SARS- CoV-2 (strain USA-WA1/2020 propagated in Vero E6 cells) diluted in assay media is added to achieve ⁇ 30 - 50% infected cells. Plates are incubated for 24 h at 34°C 5% CO2, and then fixed with 8% formaldehyde.
  • SARS- CoV-2 strain USA-WA1/2020 propagated in Vero E6 cells
  • mice survival twice a day In this model, most of the mice directly died due to the infection.
  • body weight loss clinical score is taken into account.
  • mice If the mice show a body weight loss between 20 and 25 % and become moribund (score 4). Mice are considered dead because the recovery is not possible and for ethical reason also.
  • Example 5 -7 Virus load ( Figure 3-5)
  • Lungs are collected at DI, D3 and D6 and put in Milteny tubes containing RTL buffer to RNA conservation. Lungs are homogenized before freezing. RNA is extracted using the RNeasy kit (Qiagen) according to the manufacturer's instructions.
  • the virus load is quantified by qPCR (absolute quantification); a standard curve are generated.
  • the test used targets 3 RNA regions: one coding for spike protein, one for coding 1
  • SUBSTITUTE SHEET (RULE 26) for a nucleocapsid protein and one coding for 0RF1 This test allows to avoid false positive or false negative amplification.
  • Example 8 Body weight (Figure 6)
  • Body weights are monitored once a day. every day by the same person at a fixed time.
  • Raw data are directly registered on an Excel.
  • Vero cells were seeded per well in 96-well flat bottom tissue culture plates in growth medium (MEM supplemented with 10% HI-FBS, P/S and L-Gln) and incubated overnight at 37°C and 5%CO2.
  • SUBSTITUTE SHEET (RULE 26) only and virus only wells received 100 pl of medium only. Plates were incubated at 35°C (for INFV ONLY) or 37°C and 5%CO2 for 60-minutes +/- 5 minutes.
  • Each virus strain was prepared in serum -free medium at a specific MOI of 0.01 (Table 2); for INFV ONLY, 1 pg/mL of TPCK trypsin is added to the medium. 100 pl of virus inoculum is then mixed to 100 pl of each TA concentrations, including virus only wells; 100 pl of medium only is also added to cells only wells for a final 200 pl per well. Plates were incubated at 35°C (for INFV ONLY) or 37°C and 5%CO2 for different number of days (Table 2) before cells fixation and staining.
  • TMB substrate 100 pL/well
  • Stop solution is then added to each well and optical density is read at 450nm.
  • Compound 1 and Acyclovir were diluted to 200 pM in supplemented media, and 225pl of these diluted stocks or diluent only (1% DMSO) were added in triplicate to the top raw (A) of a round bottom 96 well plate, as indicated below.
  • the virus stock (HCMV Merlin strain, 1x106 lU/ml) was diluted 5-fold with supplemented media to bring the concentration to 2x105 lU/ml. After Ih pre-treatment, media/treatment was removed from the cells and 50 pl per well of treatment from the dilution plate were retransferred to the cells in corresponding positions in the infectivity plates. 50 pl virus per well (MOI ⁇ 1) were added, except in column 11 (uninfected control), where 50 pl of supplemented media without virus were added.
  • the infected plates were washed with PBS, fixed for 30 mins with 4% formaldehyde, washed again with PBS, and stored in PBS at 4°C overnight until staining.
  • the cytotoxicity plates were treated with MTT to determine cell viability.
  • Cytotoxicity readout Cytotoxicity was detected by MTT assay following SOP-RA 006. Briefly, the MTT reagent (Sigma, M5655) was added to the cells for 2h at 37°C, 5% CO2, after which the media was removed and the precipitate solubilized with a mixture of 1 : 1 Isopropanol :DMSO for 20 minutes. The supernatant was transferred to a clean plate and signal read at 570nm.
  • TC50 values were extrapolated from the curves representing the best fit (non-linear regression analysis, variable slope) of the logarithm of compound concentration vs. the normalised percentages of cytotoxicity, using GraphPad Prism (version 9).
  • the graph display the percentage of inhibition of HCMV infection (black line) at different compound concentrations.
  • Sample values in each plate are normalized to the plate internal controls, where 100% inhibition is derived from the average of the negative control (untreated uninfected) and 0% inhibition is derived from the average of the positive control (untreated infected).
  • the x axes show compound dilutions (DM).
  • the curves represent the best fit of the logarithm of compound dilution vs. the normalized percentage of inhibition (variable slope). Cytotoxicity is displayed in gray, with values normalized to the plate internal control (untreated cells, 100% viability).

Abstract

The present invention encompasses diphenyl urea selected but not limited from the group of compounds 1 -7 for use in the treatment of virus and coronavirus infections, including COVID-19, alone or in combination with one or more additional therapeutic agents.

Description

DIPHENYL UREAS FOR THE TREATMENT OF VIRAL INFECTIONS
TECHNICAL FIELD OF THE INVENTION
[00011 Diphenyl urea is an important link structure in the design of active substance for treating cancer due to its near-perfect binding with certain acceptors and has demonstrated many activities against several human cancer cell lines. Diphenyl urea is utilized to treat cancer by inhibiting cell signaling transduction, such as RAS-RAFMEK- ERK signaling pathway and PI3K-Akt-mTOR pathway. In addition, this structure inhibits tumor cell growth by inhibiting receptor tyrosine kinases multiply, such as Vascular Endothelial Growth Factor Receptors (VEGFRs), Platelet-Derived Growth Factor Receptors (PDGFRs), Epidermal Growth Factor Receptors (EGFRs) (Yi-Cong et al., Mini-Reviews in Organic Chemistry, 16,7, 2019).
BACKGROUND OF THE INVENTION
Coronaviruses
[0002] Coronaviruses (CoVs) are positive-sense, single-stranded RNA (ssRNA) viruses of the order Nidovirales, in the family Coronaviridae . There are four sub-types of coronaviruses - alpha, beta, gamma and delta - with the Alphacoronaviruses and Betacoronaviruses infecting mostly mammals, including humans. Over the last two decades, three significant novel coronaviruses have emerged which jumped from a non-human mammal hosts to infect humans: the severe acute respiratory syndrome (SARS-CoV-1) which appeared in 2002, Middle East respiratory syndrome (MERS-CoV) which appeared in 2012, and COVID-19 (SARS-CoV-2) which appeared in late 2019. By mid-June of 2020, over 7.8 million people are known to have been infected, and over 432,000 people have died. Both numbers likely represent a significant undercount of the devastation wrought by the disease.
COVID-19
[0003] https://doi.org/10.1016/S0140-6736(20)30566-3). Fully understanding the mechanism of viral pathogenesis and immune responses triggered by SARS-CoV-2 would be extremely important in rational design of therapeutic interventions beyond antiviral treatments and supportive care. Much is still being discovered about the various ways that COVID-19 impacts the health of the people that develop it SARS-CoV-2 closely resembles SARS-CoV-1,
1
SUBSTITUTE SHEET (RULE 26) the causative agent of SARS epidemic of 2002-03 (Fung, et al, Annu. Rev. Microbiol. 2019. 73:529-57). Severe disease has been reported in approximately 15% of patients infected with SARS-CoV-2, of which one third progress to critical disease (e.g. respiratory failure, shock, or multi organ dysfunction (Siddiqi, et al, J. Heart and Lung Trans. (2020), doi: https://doi.Org/10.1016/j.healun.2020.03.012, Zhou, et al, Lancet 2020; 395: 1054-62.
[0004] Severe acute respiratory syndrome (SARS)-Corona Virus-2 (CoV-2), the etiologic agent for coronavirus disease 2019 (COVID-19), has caused a pandemic affecting almost eight million people worldwide with a case fatality rate of 2-4% as of June 2020. The virus has a high transmission rate, likely linked to high early viral loads and lack of pre-existing immunity (He, et. al, Nat Med 2020 https://doi.org/10.1038/s41591-020-0869-5). It causes severe disease especially in the elderly and in individuals with comorbidities. The global burden of COVID- 19 is immense, and therapeutic approaches are increasingly necessary to tackle the disease. Intuitive anti-viral approaches including those developed for enveloped RNA viruses like HIV- 1 (lopinavir plus ritonavir) and Ebola virus (remdesivir) have been implemented in testing as investigational drugs (Grein et al, NEJM 2020 https://doi.org/10.1056/NEJMoa2007016; Cao,et al, NEJM 2020 DOI: 10.1056/NEJMoa2001282). But given that many patients with severe disease present with immunopathology, host-directed immunomodulatory approaches are also being considered, either in a staged approach or concomitantly with antivirals (Metha, et al, The Lancet 2020; 395(10229) DOI: https://doi.org/10. 1016/80140-6736(20)30628-0, Stebbing, et al, Lancet Infect Dis 2020. https://doi.org/10.1016/S1473-3099(20)30132-8). Molnupiravir (MK-4482/EIDD-2801) is an investigational, orally administered form of a potent ribonucleoside analog that inhibits the replication of SARS-CoV-2, the causative agent of COVID-19. Molnupiravir has been shown to be active in several preclinical models of SARS- CoV-2, including for prophylaxis, treatment, and prevention of transmission (DOI: 10.1101/2021.06.17.21258639). Nafamostat, a serine protease inhibitor, has been used for the treatment of disseminated intravascular coagulation and pancreatitis. In vitro studies and clinical reports suggest its beneficial effect in the treatment of COVID-19 pneumonia (https://doi.org/10.1128/AAC.00754-20).
2
SUBSTITUTE SHEET (RULE 26) [0005] To date, treatment typically consists only of the available clinical mainstays of symptomatic management, oxygen therapy, with mechanical ventilation for patients with respiratory failure. Thus, there is an urgent need for novel therapies to address the different stages of the SARS-CoV-2 infectious cycle (Siddiqi, et al.).
Diphenyl urea inhibitbitors with antiviral activity against RNA viruses
[0006] Viruses take over a large number of host kinases at distinct steps of their life cycle thus kinases represent an attractive target for broad-spectrum therapy [1] in general but it is unclear what is the best combination of kinases to be inhibited. Garcia et al [doi: https://doi.org/10.1101/2020.06.24.150326] mention mTOR-P 13K-AKT, ABL-25 BCR/MAPK and DNA Damage Response. Several other publication mention JAK inhibitors. JAK inhibitors have shown significant benefit and have been used in treating several autoimmune diseases and inflammatory diseases, like rheumatoid arthritis and inflammatory bowl disease. In a systematic review and meta-analysis it was indicated that JAK-inhibitors showed favorable outcome in Covid- 19, in terms of recovery rate, shortened time to recovery, reduction risk of clinical deterioration, and reduction of mortality rate [Limen et al., Expert Review of Anti-infective Therapy https://doi.org/10.1080/14787210.2021.1982695]. Another publication analyzed reports on safety and efficacy of JAK-inhibitors and found that they decreased the use of invasive mechanical ventilation and increased survival [Chen et al., Leukemia 2021, 35:2616-2620, https://doi.org/10.1038/s41375-021-01266-6]. Also, VEGF is deemed a promising therapeutic target in suppressing inflammation during SARS-CoV2 infection with neurological symptoms [Yin et al., 2020, ACS Chem Neurosci 2020, 11, 12, https://doi.org/10.1021/acschemneuro.0c00294. And Zambrana et al. [2021, Scientific Reports 11: 18985, https://doi.org/10.1038/s41598-021-98289-x] found that NO and VEGF signaling are potential molecular pathways whose function are very similar with several observed COVID-19 symptoms. Related to VEGF-signaling we suggest as a putative target gene KDR (VEGFR-2).
[0007] The scaffolds base on diphenyl urea motive showed in a cellular infection model antiviral activity against SARS-CoV-2 (Figure 15). In an in vivo mouse model Compound 7
3
SUBSTITUTE SHEET (RULE 26) demonstrated a higher percentage of survival in comparison to vehicle and a significant reduced virus load.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1
[0008] Diphenly urea scaffold with the following side chains: R1 = mono-, di- and trisubstituted phenyl, heteroaryl-phenyl, morpholino -phenyl and combinations therof with heteroaryl = substituted and unsubstitued pyrrols, pyrrazoles, imidazoles, 1,2,4 -triazoles; R2 = H, -CO-NHR, -NH-CO-OR with R = alkyl
Figure 2
[0009] Survival proportions in a SARS-CoV-2 pulmonary infection mouse model treated with Compound 7 in comparison to vehicle and positive control Molnupiravir.
Figure 3
[0010] Reduction of Virus load in a SARS-CoV-2 pulmonary infection mouse model at day 1 treated with Compound 7 in comparison to vehicle and the positive control Molnupiravir. N = 5, **P<0.01. ANOVA, followed by Dunnett’s Multi Comparison test.
Figure 4
SUBSTITUTE SHEET (RULE 26) [0011] Reduction of Virus load in a SARS-CoV-2 pulmonary infection mouse model at day 3 treated with Compound 7 in comparison to vehicle and the positive control Molnupiravir. N = 5, *P<0.05, ***P<0.001. ANOVA, followed by Dunnett’s Multi Comparison test.
Figure 5
[0012] Reduction of Virus load in a SARS-CoV-2 pulmonary infection mouse model at day 6 treated with Compound 7 in comparison to vehicle and the positive control Molnupiravir. N = 5, ***P<0.001. ANOVA, followed by Dunnett’s Multi Comparison test.
Figure 6
[0013] Body weight loss in a SARS-CoV-2 pulmonary infection mouse model over ten days in mice treated with Compound 7 in comparison to vehicle and the positive control Molnupiravir. N = 10
Figure 7
[0014] Neutralization of Compound 1 against Adenovirus-5 in Vero cells on day 4. IC50=
1.164pM.
Figure 8
[0015] Neutralization of Compound 7 against Adenovirus-5 in Vero cells on day 3. IC50=
0.6291pM
Figure 9
[0016] Neutralisation of Compound 7 against Dengue Vims in Vero cells on day 3
5
SUBSTITUTE SHEET (RULE 26) Figure 10
[0017] Neutralization of Compound 1 against Dengue Virus in Vero cells on day 4. IC50= 8.434
Figure 11
[0018] Neutralization of Compound lagainst Herpes simplex Virus-subtype 2 in Vero cells on day 4. IC50 = 5.224 pM.
Figure 12
[0019] Neutralization of Compound 7 against Herpes simplex Virus-subtype 2 in Vero cells on day 3. IC50= 5.224 pM.
Figure 13
[0020] Percentage of inhibition of HCMV infection of different Compound 1 concentrations in human foreskin fibroblasts (HFF) on day 5. IC50= 8.255 pM.
Figure 14
[0021] Antiviral Activity of Compound 7 on Hela3-ACE2 cells
Figure 15
[0022] IC50 determination of compounds with diphenyl urea motive in Calu-3 and HeLa-ACE2 SARS CoV-2 assays. SI = safety index (average CC50 / average EC 50)
6
SUBSTITUTE SHEET (RULE 26)
Summary of the Invention
[0023] In a first embodiment, the invention provides compounds 1 -7 for use in the treatment of viral infections in a subject in need thereof. The compounds are subject matter of W02006040056. In one aspect of this embodiment, the viral infection is a single-strand RNA viral infection. In another aspect of this embodiment, the viral infection is a coronavirus infection. In a further aspect of this embodiment, the viral infection is a SARS-CoVl, MERS- CoV, or SARS-CoV-2 infection. In a final aspect of this embodiment, the viral infection is a SARS-CoV-2 infection.
[0024] A second embodiment is a method of treating a coronavirus infection in a subject in need thereof, comprising administering an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7, or a pharmaceutically acceptable salt thereof, to the subject. In one aspect of this embodiment, the administration of a diphenyl urea selected but not limited from the group of compounds 1 -7 reduces the viral load in the subject. In one aspect of this embodiment, the subject has a mild to moderate SARS-CoV-2 infection. In an additional aspect of this embodiment, the subject is asymptomatic at the start of the administration regimen.
1. Compounds and Definitions
SUBSTITUTE SHEET (RULE 26)
Figure imgf000009_0001
R1 = mono-, di- and tri-substituted phenyl, heteroaryl -phenyl, morpholino -phenyl and combinations therof with heteroaryl = substituted and unsubstitued pyrrols, pyrrazoles, imidazoles, 1,2,4 -triazoles
R2 = H, -CO-NHR, -NH-CO-OR with
R = alkyl
Compound 1
4-{4-[3-(2-[l,2,4] Triazol-l-yl-5-trifluoromethyl-phenyl)-ureido]-phenoxy}-pyridine-2- carboxylic acid methylamide
Figure imgf000009_0002
SUBSTITUTE SHEET (RULE 26) Compound 2
4-{4-[3-(2-Imidazol-l-yl-5-trifluoromethyl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide
Figure imgf000010_0001
Compound 3 l-[4-(Pyridin-4-yloxy)-phenyl]-3-(2-[l,2,4] triazol-l-yl-5-trifluoromethyl-phenyl)-urea
Figure imgf000010_0002
Compound 4
(5-{4-[3-(2-Fluoro-5-trifluoromethyl-phenyl)-ureido]-phenoxy}-lH-benzoimidazol-2-yl)- carbamic acid methyl ester
9
SUBSTITUTE SHEET (RULE 26)
Figure imgf000011_0001
Compound 5
4-{4-[3-(5-tert-Butyl-2-m-tolyl-2H-pyrazol-3-yl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide
Figure imgf000011_0002
Compound 6
4-{4-[3-(2-Morpholin-4-yl-5-trifluoromethyl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide
Figure imgf000011_0003
10
SUBSTITUTE SHEET (RULE 26) Compound 7
4-{4-[3-(4-Chloro-5-methyl-2-pyrrol-l-yl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide
Figure imgf000012_0001
DETAILED DESCRIPTION
[0025] At the initial antiviral response phase, when the virus primarily infects ACE2- expressing specialized epithelial cells (type II pneumocytes) in the lung alveoli, direct anti-viral or immune-enhancing therapy (e.g. interferon-beta-1) may prove to be of benefit in minimizing contagion and preventing progression to severe disease (Hoffmann, et al, Cell 2020. DOI: https://doi.Org/10.1016/j.cell.2020.02.052; Sungnak, et al, Qbio preprint; arXiv:2003.06122 [q- bio.CB]; Zou, et al, Front Med 2020 https://doi.org/10.1007/sl l684-020-0754-0; Zhao, et al, BioRxv preprint https://doi.org/10.1101/2020.01.26.919985; Qi, et al, BBRC 2020 https://doi.Org/10.1016/j.bbrc.2020.03.044; Taccone, et al, Lancet Resp. Med. (2020) https ://doi . org/ 10.1016/S2213 -2600(20)30172-7) .
[0026] Indeed, recent papers have suggested a correlation between SARS-CoV-2 viral load, symptom severity and viral shedding (He, et al; Liu, et al, Lancet Infect Dis 2020. https://doi.org/10.1016/S1473-3099(20)30232-2). Some antiviral drugs administered at symptom onset to blunt coronavirus replication are in the testing phase (Grein, et al; Taccone, et al), but as yet none have shown much promise.
[0027] SARS-CoV-2 directly enters cells expressing ACE2 via receptor-mediated endocytosis (Hoffmann, et al). Successful viral replication requires host endosome acidification
11
SUBSTITUTE SHEET (RULE 26) to release the viral genome into the host cytosol. Innate immune cells like monocytes, macrophages and neutrophils do not highly express ACE2, but have abundant Fc receptors (Zou, et al; Qi, et al; Lu, et al, Nat. Rev. Imm. 2018 https://doi.org/10.1038/nri.2017.106). In stage II (Fig.l), antibodies that bind the virus, can mediate viral uptake into myeloid cell endosomes via Fc receptors (FcR) or complement receptors (CR) (Lu, et al; Dandekar, et al, Nat. Rev. Imm. 2005, https://doi.org/10.1038/nri l732). Thus ACE2, FcR and CR present three mechanisms how SARS-CoV-2 can enter endosomes and trigger hyperinflammation leading to cytokine storm and severe disease. Additionally, ssRNA virus can induce NETosis in neutrophils (Saitoh, et al, Cell Host Microbe (2012), 19;12(1): 109-16) leading to release of DNA and RNA, creating a feed-forward loop to further fuel inflammation (Herster et al, Nat Commun 2020; 11, 105 https://doi.org/10.1038/s41467-019-13756-4), which has been proposed as a driver of severe COVID-19 (Barnes, et al, I Exp med 2020; 217 (6) https://doi.org/10.1084/jem.20200652). SARS-CoV-1 derived ssRNA has been shown to mediate severe lung pathology in animal models and presents as a potential driver of virus- associated cytokine storm (Li, et al, Microbes Infect 2013; 15 (2) 88-95. https://doi.Org/10.1016/j.micinf.2012.10.008). Being able to slow the viral reproduction in the early stages of infection may allow the subject to avoid severe disease.
[0028] COVID-19” is the name of the disease which is caused by a SARS-CoV-2 infection. While care was taken to describe both the infection and disease with accurate terminology, “COVID-19” and “SARS-CoV-2 infection” are meant to be roughly equivalent terms.
[0029] As of the writing of this application, the determination and characteristics of the severity of COVID-19 patients/symptoms has not been definitively established. However, in the context of this invention, “mild to moderate” COVID-19 occurs when the subject presents as asymptomatic or with less severe clinical symptoms (e.g., low grade or no fever (<39.1°C), cough, mild to moderate discomfort) with no evidence of pneumonia, and generally does not require medical attention. When “moderate to severe” infection is referred to, generally patients present with more severe clinical symptoms (e.g., fever >39.1°C, shortness of breath, persistent cough, pneumonia, etc.). As used herein “moderate to severe” infection typically requires
12
SUBSTITUTE SHEET (RULE 26) medical intervention, including hospitalization. During the progression of disease, a subject can transition from “mild to moderate” to “moderate to severe” and back again in one course of bout of infection.
[0030] Treatment of CO VID-19 using the methods of this invention include administration of an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention at any stage of the infection to prevent or reduce the symptoms associated therewith. Typically, subjects will be administered an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention after definitive diagnosis and presentation with symptoms consistent with a SARS-CoV2 infection, and administration will reduce the severity of the infection and/or prevent progression of the infection to a more severe state. The clinical benefit upon such administration is described in more detail in the sections below.
[0031] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.
[0032] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e g , enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention.
[0033] Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13C- or 14C-enriched carbon are
13
SUBSTITUTE SHEET (RULE 26) within the scope of this invention. In some embodiments, the group comprises one or more deuterium atoms.
2. Uses, Formulation and Administration
[0034] The term “patient” or “subject”, as used herein, means an animal, preferably a human. However, “subject” can include companion animals such as dogs and cats. In one embodiment, the subject is an adult human patient. In another embodiment, the subject is a pediatric patient. Pediatric patients include any human which is under the age of 18 at the start of treatment. Adult patients include any human which is age 18 and above at the start of treatment. In one embodiment, the subject is a member of a high-risk group, such as being over 65 years of age, immunocompromised humans of any age, humans with chronic lung conditions (such as, asthma, COPD, cystic fibrosis, etc.), and humans with other co-morbidities. In one aspect of this embodiment, the other co-morbidity is obesity, diabetes, and/or hypertension.
[0035] Compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Preferably, the compositions are administered orally. In one embodiment, the oral formulation of a compound of the invention
[0036] is a tablet or capsule form. In another embodiment, the oral formulation is a solution or suspension which may be given to a subject in need thereof via mouth or nasogastric tube. Any oral formulations of the invention may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.
[0037] Pharmaceutically acceptable compositions of this invention are orally administered in any orally acceptable dosage form. Exemplary oral dosage forms are capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also
14
SUBSTITUTE SHEET (RULE 26) typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents are optionally also added.
[0038] The amount of compounds of the present invention that are optionally combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of between 0.1 - 100 mg/kg body weight/day of the compound can be administered to a patient receiving these compositions.
[0039] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 of the invention act as host-targeting antivirals by blocking the host pyrimidine synthesis, and thereby blocking the viral replication.
[0040] In one embodiment, the total amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 administered to the subject in need thereof is generally in the range from 0.2 to 90 mg/kg of body weight of the recipient (mammal) per day and particularly typically in the range from 1 to 10 mg/kg of body weight per day. Thus, the actual amount per day for an adult mammal weighing 70 kg is usually between 70 and 700 mg, where this amount can be administered as an individual dose per day or usually in a series of part-doses (such as, for example, two, three, four, five or six) per day, so that the total daily dose is the same. An effective amount of a salt or solvate or of a physiologically functional derivative of the diphenyl urea selected but not limited from the group of compounds 1 -7 can be determined as the fraction of the effective amount of the compound per se.
[0041] In the above embodiment, diphenyl urea selected but not limited from the group of compounds 1 -7 are administered for a period of about 7 days to about 28 days. In one aspect of any of the above embodiments, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered for about 14 days.
[0042] In one embodiment of the invention, the subject is suffering from COVID-19 pneumonia. In one embodiment of this invention, the subject is suffering from one or more
15
SUBSTITUTE SHEET (RULE 26) symptoms selected from chest congestion, cough, blood oxygen saturation (SpCh) levels below 94%, shortness of breath, difficulty breathing, fever, chills, repeated shaking with chills, muscle pain and/or weakness, headache, sore throat and/or new loss of taste or smell.
[0043] In one embodiment of the invention, the subject is being treated inpatient in a hospital setting. In another embodiment, the subject is being treated in an outpatient setting. In one aspect of the preceding embodiments, the subject may continue administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are after being transitioned from being treated from an inpatient hospital setting to an outpatient setting.
[0044] In one embodiment, the administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are results in one or more clinical benefit. In one aspect of this embodiment, the one or more clinical benefit is selected from the group comprising: reduction of duration of a hospital stay, reduction of the duration of time in the Intensive Care Unit (ICU), reduction in the likelihood of the subject being admitted to an ICU, reduction in the rate of mortality, reduction in the likelihood of kidney failure requiring dialysis, reduction in the likelihood of being put on non-invasive or invasive mechanical ventilation, reduction of the time to recovery, reduction in the likelihood supplemental oxygen will be needed, improvement or normalization in the peripheral capillary oxygen saturation (SpCh levels) without mechanical intervention, reduction of severity of the pneumonia as determined by chest imaging (eg, CT or chest X ray), reduction in the cytokine production, reduction of the severity of acute respiratory distress syndrome (ARDS), reduction in the likelihood of developing ARDS, clinical resolution of the COVID-19 pneumonia, improvement of the PaO2/FiO2 ratio, and reduction of the inflammatory response in the subject.
[0045] In another embodiment, the one or more clinical benefits includes the improvement or normalization in the peripheral capillary oxygen saturation (SpO2 levels) in the subject without mechanical ventilation or extracorporeal membrane oxygenation.
[0046] In a further embodiment, the one of more clinical benefits is reduction in the likelihood of being hospitalized, reduction in the likelihood of ICU admission, reduction in the likelihood being intubated (invasive mechanical ventilation), reduction in the likelihood supplemental oxygen will be needed, reduction in the length of hospital stay, reduction in the
16
SUBSTITUTE SHEET (RULE 26) likelihood of mortality, and/or a reduction in likelihood of relapse, including the likelihood of rehospitalization.
[0047] The invention also provides a method of treating a viral infection caused by F Herpes simplex virus (HSV), human cytomegalo virus (HCMV), Dengue virus, zoonotic pandemic viruses and infections by coronaviruses in a subject in need thereof comprising administering an effective amount of a compound of the invention to the subject. An amount effective to treat or inhibit a viral infection is an amount that will cause a reduction in one or more of the manifestations of viral infection, such as viral lesions, viral load, rate of virus production, and mortality as compared to untreated control subjects.
[0048] One embodiment of the invention is a method of treating a coronavirus infection in a subject in need thereof, comprising administering an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7 are, or a pharmaceutically acceptable salt thereof, to the subject. In one aspect of this embodiment, the subject is infected with Severe acute respiratory -related coronavirus (SARS-CoV-1, SARS-CoV-2) and Middle East Syndrome coronavirus (MERS). In another aspect of this embodiment, the administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are results in the reduction of the viral load in the subject.
[0049] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to COVID-19 pneumonia developing. In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to the subject developing a cytokine storm Tn another embodiment, the subject has a mild to moderate SARS-CoV-2 infection. In a further embodiment, the subject is asymptomatic at the start of the administration regimen. In another embodiment, the subject has had known contact with a patient who has been diagnosed with a SARS-CoV-2 infection. In an additional embodiment, the subject begins administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 are prior to being formally diagnosed with COVID- 19.
[0050] In one aspect of this embodiment, the one or more clinical benefits is shortening the duration of infection, reduction of the likelihood of hospitalization, reduction in the likelihood
17
SUBSTITUTE SHEET (RULE 26) of mortality, reduction in the likelihood of ICU admission, reduction in the likelihood being placed on mechanical ventilation, reduction in the likelihood supplemental oxygen will be needed, and/or reduction in the length of hospital stay. In another aspect of this embodiment, the one or more clinical benefits is avoidance of a significant proinflammatory response. In a further aspect of this embodiment, the one or more clinical benefit is the failure of the subject to develop significant symptoms of COVTD-19.
[0051] The compounds of the invention can be administered before or following an onset of SARS-CoV-2 infection, or after acute infection has been diagnosed in a subject. A therapeutically relevant effect relieves to some extent one or more symptoms of a disorder, or returns to normality, either partially or completely, one or more physiological or biochemical parameters associated with or causative of a disease or pathological condition. Monitoring is considered as a kind of treatment provided that the compounds are administered in distinct intervals, e.g. in order to boost the response and eradicate the pathogens and/or symptoms of the disease. The methods of the invention can also be used to reduce the likelihood of developing a disorder or even prevent the initiation of disorders associated with COVID-19 in advance of the manifestation of mild to moderate disease, or to treat the arising and continuing symptoms of an acute infection.
[0052] Treatment of mild to moderate CO VID-19 is typically done in an outpatient setting. Treatment of moderate to severe COVID-19 is typically done inpatient in a hospital setting. Additionally, treatment can continue in an outpatient setting after a subject has been discharged from the hospital.
[0053] The invention furthermore relates to a medicament comprising at least one diphenyl urea selected but not limited from the group of compounds 1 -7 are according to the invention, or a pharmaceutically salts thereof.
[0054] A “medicament” in the meaning of the invention is any agent in the field of medicine, which comprises one or more compounds of the invention or preparations thereof (e.g. a pharmaceutical composition or pharmaceutical formulation) and can be used in prophylaxis, therapy, follow-up or aftercare of patients who suffer from clinical symptoms and/or known exposure to COVID- 19.
18
SUBSTITUTE SHEET (RULE 26) Combination Treatment
[0055] In various embodiments, the active ingredient may be administered alone or in combination with one or more additional therapeutic agents. A synergistic or augmented effect may be achieved by using more than one compound in the pharmaceutical composition. The active ingredients can be used either simultaneously or sequentially.
[0056] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more additional therapeutic agents. In one aspect of this embodiment, the one or more additional therapeutic agents is selected from anti-inflammatories, antibiotics, anti-coagulants, antiparasitic agent, antiplatelet agents and dual antiplatelet therapy, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, beta-blockers, statins and other combination cholesterol lowering agents, specific cytokine inhibitors, complement inhibitors, anti-VEGF treatments, JAK inhibitors, BTK inhibitors, TLR(7/8) inhibitors, immunomodulators, anti-inflammasone therapies, sphingosine-1 phosphate receptors binders, N-methyl-d-aspartate (NDMA) receptor glutamate receptor antagonists, corticosteroids, Granulocyte-macrophage colony-stimulating factor (GM-CSF), anti-GM-CSF, interferons, angiotensin receptor-neprilysin inhibitors, calcium channel blockers, vasodilators, diuretics, muscle relaxants, and antiviral medications.
[0057] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with another antiviral agent. In one aspect of this embodiment, the antiviral agent is remdesivir. In another aspect of this embodiment, the antiviral agent is lopinavir-ritonavir, alone or in combination with ribavirin and interferon-beta.
[0058] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administrated in combination with a broad-spectrum antibiotic.
[0059] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 is is administered in combination with chloroquine or hydroxychloroquine. In one aspect of this embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 is is further combined with azithromycin.
19
SUBSTITUTE SHEET (RULE 26) [0060] In one embodiment, the diphenyl urea selected but not limited from the group of compound 1 -7 is is administered in combination with one or more additional therapeutic agents selected from hydroxychloroquine, chloroquine, ivermectin, tranexamic acid, nafamostat, virazole, ribavirin, lopinavir/ritonavir, favipiravir, arbidol, leronlimab, interferon beta- la, interferon beta- lb, beta-interferon, azithromycin, nitr zoxamide, lovastatin, clazakizumab, adalimumab, etanercept, golimumab, infliximab, sarilumab, tocilizumab, anakinra, emapalumab, pirfenidone, belimumab, rituximab, ocrelizumab, anifrolumab, ravulizumab- cwvz, eculizumab, bevacizumab, heparin, enoxaparin, apremilast, coumadin, baricitinib, ruxolitinib, acalabrutinib, dapafliflozin, methotrexate, azathioprine, sulfasalazine, mycophenolate mofetil, colchicine, fingolimod, ifenprodil, prednisone, cortisol, dexamethasone, methylprednisolone, melatonin, otilimab, ATR-002, APN-01, camostat mesylate, brilacidin, IFX-1, PAX-1-001, BXT-25, NP-120, intravenous immunoglobulin (IVIG), and solnatide.
[0061] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more anti-inflammatory agent. In one aspect of this embodiment, the anti-inflammatory agent is selected from corticosteroids, steroids, COX-2 inhibitors, and non-steroidal anti-inflammatory drugs (NSAID). In one aspect of this embodiment, the anti-inflammatory agent is diclofenac, etodolac, fenoprofen, flurbirprofen, ibuprofen, indomethacin, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, sulindac, tolmetin, celecoxib, prednisone, hydrocortisone, fludocortisone, bethamethasone, prednisolone, triamcinolone, methylprednisone, dexamethasone, fluticasone, and budesonide (alone or in combination with formoterol, salmeterol, or vilanterol).
[0062] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more immune modulators. In one aspect of this embodiment the immune modulator is a calcineurin inhibitor, antimetabolite, or alkylating agent. In another aspect of this embodiment, the immune modulator is selected from azathioprine, mycophenolate mofetil, methotrexate, dapson, cyclosporine, cyclophosphamide, and the like.
20
SUBSTITUTE SHEET (RULE 26) [0063] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antibiotics. In one aspect of this embodiment, the antibiotic is a broad-spectrum antibiotic. In another aspect of this embodiment, the antibiotic is a penicillin, anti-staphylococcal penicillin, cephalosporin, aminopenicillin (commonly administered with a betalactamase inhibitor), monobactam, quinoline, aminoglycoside, lincosamide, macrolide, tetracycline, glycopeptide, antimetabolite or nitroimidazole. In a further aspect of this embodiment, the antibiotic is selected from penicillin G, oxacillin, amoxicillin, cefazolin, cephalexin, cephotetan, cefoxitin, ceftriazone, augmentin, amoxicillin, ampicillin (plus sulbactam), piperacillin (plus tazobactam), ertapenem, ciprofloxacin, imipenem, meropenem, levofloxacin, moxifloxacin, amikacin, clindamycin, azithromycin, doxycycline, vancomycin, Bactrim, and metronidazole.
[0064] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more anti-coagulants. In one aspect of this embodiment, the anti-coagulant is selected from apixaban, dabigatran, edoxaban, heparin, rivaroxaban, and warfarin.
[0065] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antiplatlet agents and/or dual antiplatelet therapy. In one aspect of this embodiment, the antiplatelet agent and/or dual antiplatelet therapy is selected from aspirin, clopidogrel, dipyridamole, prasugrel, and ticagrelor.
[0066] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more ACE inhibitors. In one aspect of this embodiment, the ACE inhibitor is selected from benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril and trandolapril.
[0067] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more angiotensin II receptor blockers. In one aspect of this embodiment, the angiotensin II receptor blocker is selected from azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan.
21
SUBSTITUTE SHEET (RULE 26) [0068] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more beta-blockers. In one aspect of this embodiment, the beta-blocker is selected from acebutolol, atenolol, betaxolol, bisoprolol/hydrochlorothiazide, bisoprolol, metoprolol, nadolol, propranolol, and sotalol.
[0069] In another embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more alpha and beta-blocker. In one aspect of this embodiment, the alpha and beta-blocker is carvedilol or labetalol hydrochloride.
[0070] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more interferons, such as interferon-beta. In an aspect, the interferon-beta is interferon-beta-1. In a specific aspect, the interferon-beta is interferon-beta- la.
[0071] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more angiotensin receptor- neprilysin inhibitors. In one aspect of this embodiment, the angiotensin receptor-neprilysin inhibitor is sacubitril/valsartan.
[0072] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more calcium channel blockers. In one aspect of this embodiment, the calcium channel blocker is selected from amlodipine, diltiazem, felodipine, nifedipine, nimodipine, nisoldipine, and verapamil.
[0073] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more vasodilators. In one aspect of this embodiment, the one or more vasodilator is selected from isosorbide dinitrate, isosorbide mononitrate, nitroglycerin, and minoxidil.
[0074] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more diuretics. In one aspect of this embodiment, the one or more diuretics is selected from acetazolamide, amiloride,
22
SUBSTITUTE SHEET (RULE 26) bumetanide, chlorothiazide, chlorthalidone, furosemide, hydrochlorothiazide, indapamide, metalozone, spironolactone, and torsemide.
[0075] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more muscle relaxants. In one aspect of this embodiment, the muscle relaxant is an antispasmodic or antispastic. In another aspect of this embodiment, the one or more muscle relaxants is selected from casisoprodol, chlorzoxazone, cyclobenzaprine, metaxalone, methocarbamol, orphenadrine, tizanidine, baclofen, dantrolene, and diazepam.
[0076] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more antiviral medications.
[0077] In one embodiment, the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered in combination with one or more additional therapeutic agents selected from antiparasitic drugs (including, but not limited to, hydroxychloroquine, chloroquine, ivermectin), antivirals (including, but not limited to, tranexamic acid, nafamostat, virazole [ribavirin], lopinavir/ritonavir, favipiravir, leronlimab, interferon beta-la, interferon beta-lb, beta-interferon), antibiotics with intracellular activities (including, but not limited to azithromycin, nitrazoxamide), statins and other combination cholesterol lowering and antiinflammatory drugs (including, but not limited to, lovastatin), specific cytokine inhibitors (including, but not limited to, clazakizumab, adalimumab, etanercept, golimumab, infliximab, sarilumab, tocilizumab, anakinra, emapalumab, pirfenidone), complement inhibitors (including, but not limited to, ravulizumab-cwvz, eculizumab), anti-VEGF treatments (including, but not limited to, bevacizumab), anti-coagulants (including, but not limited to, heparin, enoxaparin, apremilast, coumadin), JAK and BTK inhibitors, TLR(7/8) inhibitors, anti-inflammasone therapies (including, but not limited to, colchicine), sphingosine- 1 phosphate receptors binders (including, but not limited to, fmgolimod), N-methyl-d-aspartate (NDMA) receptor glutamate receptor antagonists (including, but not limited to, ifenprodil), corticosteroids (including, but not limited to, prednisone, cortisol, dexamethasone, methylprednisolone), GM-CSF, anti-GM-CSF (otilimab), ATR-002, APN-01, camostat
23
SUBSTITUTE SHEET (RULE 26) mesylate, arbidol, brilacidin, IFX-1, PAX-1-001, BXT-25, NP-120, intravenous immunoglobulin (IVIG), and solnatide.
[0078] In some embodiments, the diphenyl urea selected but not limited from the group of compounds 1 -7 with one or more additional therapeutic agents reduces the effective amount (including, but not limited to, dosage volume, dosage concentration, and/or total drug dose administered) of diphenyl urea selected but not limited from the group of compounds 1 -7 and/or the one or more additional therapeutic agents administered to achieve the same result as compared to the effective amount administered when the diphenyl urea selected but not limited from the group of compounds 1 -7 or the additional therapeutic agent is administered alone. In some embodiments, the combination of a diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent reduces the total duration of treatment compared to administration of the additional therapeutic agent alone. In some embodiments, the combination of a diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent reduces the side effects associated with administration of the additional therapeutic agent alone. In some embodiments, the combination of an effective amount of the diphenyl urea selected but not limited from the group of compounds 1 -7 with the additional therapeutic agent is more efficacious compared to an effective amount of the diphenyl urea selected but not limited from the group of compounds 1 -7 or the additional therapeutic agent alone. In one embodiment, the combination of an effective amount of the dual diphenyl urea selected but not limited from the group of compounds 1 -7 with the one or more additional therapeutic agent results in one or more additional clinical benefits than administration of either agent alone.
[0079] As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a viral infection, or one or more symptoms thereof, as described herein. In some embodiments, treatment is administered after one or more symptoms have developed. In other embodiments, treatment is administered in the absence of symptoms. For example, treatment is administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a known exposure to an infected person and/or in light of comorbidities which are predictors for severe disease, or other susceptibility factors).
24
SUBSTITUTE SHEET (RULE 26) 3. Exemplification
[0080] As depicted in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present invention, the following general methods, and other methods known to one of ordinary skill in the art, can be applied to all compounds and subclasses and species of each of these compounds, as described herein.
Example 1: Synthesis of Compounds C 1-7
[0081] Compound 1 -7 can be synthesized as described W02006040056, page 189, Synthesis of ureas.
Example 2: SARS-CoV-2/Calu-3 high-content screening assay (Figure 14 and Figure 15)
[0082] Compounds are acoustically transferred into 384-well pci ear-bottom plates (Greiner, Part. No. 781090-2B) before seeding Calu-3 cells in assay media (MEM with 2% FBS) at a density of 5,000 cells per 20 pL per well. The plated cells are transported to the BSL3 facility where SARS-CoV-2 (strain USA-WA1/2020 propagated in Vero E6 cells) diluted in assay media is added at an MOI between 0.75 and 1 to achieve ~30 - 60% infected cells. Plates are incubated for 48 h at 34°C 5% CO2, and then fixed with a final concentration of 4% formaldehyde. Fixed cells are stained with human polyclonal sera as the primary antibody, goat anti-human H+L conjugated Alexa 488 (Thermo Fisher Scientific Al 1013) as the secondary antibody, and antifade-46-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific D1306) to stain DNA, with PBS 0.05% Tween 20 washes in between fixation and subsequent primary and secondary antibody staining.
[0083] Plates are imaged using the ImageXpress Micro Confocal High-Content Imaging System (Molecular Devices) with a 10x objective, with 4 fields imaged per well. Images are analyzed using the Multi -Wavelength Cell Scoring Application Module (MetaXpress), with DAPI staining identifying the host-cell nuclei (the total number of cells in the images) and the SARS-CoV-2 immunofluorescence signal leading to identification of infected cells.
Uninfected host cell cytotoxicity counter screen
25
SUBSTITUTE SHEET (RULE 26) [0084] Compounds are acoustically transferred into 1,536-well plates (Corning No. 9006BC) before seeding Calu-3 cells in assay media (MEM with 2% FBS) at a density of 600 cells per 5 pL per well. Plates are incubated for 48 hours at 37°C 5% CO2. To assess cell viability, 2 L of 50% Cell-Titer Gio (Promega No G7573) diluted in water is added to the cells and luminescence measured on an EnVision Plate Reader (Perkin Elmer)
Data analysis
[0085] Data from the SARS-CoV-2 antiviral assay and host cell cytotoxicity counter screen are uploaded to Genedata Screener, Version 16.0. For the SARS-CoV-2 antiviral readout, the % CoV-2 positive cells are normalized to neutral (DMSO) minus inhibitor controls (10 pM remdesivir). For the cell count readout, the total cells are normalized to the stimulator (10 pM remdesivir) minus neutral control (DMSO). The uninfected host cell cytotoxicity counter screen is normalized to neutral (DMSO) minus inhibitor control (30 pM puromycin). For dose response experiments, compounds are tested in technical triplicates on different assay plates and dose curves are fitted with the four parameter Hill Equation. Curves are fitted as either increasing or decreasing and noted as such in the data output. This is of particular note for the cell count readout from the SARS-CoV-2 infection assay which captures both an antiviral effect, protection from virus-induced cell death (increasing), and cellular toxicity (decreasing).
Example 3: SARS-CoV-2/HeLa-ACE2 high-content screening assay (Figure 15)
[0086] Compounds are acoustically transferred into 384-well pclear-bottom plates (Greiner, Part. No. 781090-2B) and HeLa-ACE2 cells are seeded in the plates in 2% FBS at a density of 1.0x 103 cells per well. Plated cells are transported to the BSL3 facility where SARS- CoV-2 (strain USA-WA1/2020 propagated in Vero E6 cells) diluted in assay media is added to achieve ~30 - 50% infected cells. Plates are incubated for 24 h at 34°C 5% CO2, and then fixed with 8% formaldehyde. Fixed cells are stained with human polyclonal sera as the primary antibody, goat anti-human H+L conjugated Alexa 488 (Thermo Fisher Scientific Al 1013) as the secondary antibody, and antifade-46-diamidino-2-phenylindole (DAPI; Thermo Fisher
26
SUBSTITUTE SHEET (RULE 26) Scientific D1306) to stain DNA, with PBS 0.05% Tween 20 washes in between fixation and subsequent primary and secondary antibody staining.
Example 4: Survival (Figure 2)
[0087] We monitored mice survival twice a day. In this model, most of the mice directly died due to the infection.
Furthermore, concerning some mice, other criteria are taken into account: body weight loss clinical score
1 : healthy mouse
2: mouse showing signs of malaise, including slight piloerection, slightly changed gait and increased ambulation
3 : mouse showing signs of strong piloerection, constricted abdomen, changed gait, periods of inactivity
4: mouse with enhanced characteristics of the previous group, but showing little activity and becoming moribund
5: If the mice show a body weight loss between 20 and 25 % and become moribund (score 4). Mice are considered dead because the recovery is not possible and for ethical reason also.
Example 5 -7: Virus load (Figure 3-5)
[0088] Lungs are collected at DI, D3 and D6 and put in Milteny tubes containing RTL buffer to RNA conservation. Lungs are homogenized before freezing. RNA is extracted using the RNeasy kit (Qiagen) according to the manufacturer's instructions.
[0089] The virus load is quantified by qPCR (absolute quantification); a standard curve are generated. The test used targets 3 RNA regions: one coding for spike protein, one for coding 1
SUBSTITUTE SHEET (RULE 26) for a nucleocapsid protein and one coding for 0RF1 This test allows to avoid false positive or false negative amplification.
Positive and negative control are included in the tests.
Example 8 Body weight (Figure 6)
[0090] Body weights are monitored once a day. every day by the same person at a fixed time. Raw data are directly registered on an Excel.
Example 9 -13 Efficacy Graph for Cytotoxicity (Figures 7, 10 and 11 Compound 1; Figures 8,9 and 12 Compound 7)
CPE-based Assay
Cell Seeding
[0091] 10,000 Vero cells were seeded per well in 96-well flat bottom tissue culture plates in growth medium (MEM supplemented with 10% HI-FBS, P/S and L-Gln) and incubated overnight at 37°C and 5%CO2.
TA Formulation
[0092] Eight 5-fold serial dilutions of each TA (Figure 15) was prepared in serum-free medium at two times (2X) the final intended concentration; for INFV ONLY, 1 pg/mL of TPCK trypsin is added to the medium.
Cell/TA Incubation
[0093] Growth medium from the 96-well plates were removed; 50 pL fresh serum-free medium was added to each well followed by 50 pL of each TA dilutions in triplicates; cells
28
SUBSTITUTE SHEET (RULE 26) only and virus only wells received 100 pl of medium only. Plates were incubated at 35°C (for INFV ONLY) or 37°C and 5%CO2 for 60-minutes +/- 5 minutes.
Virus dilution
[0094] Each virus strain was prepared in serum -free medium at a specific MOI of 0.01 (Table 2); for INFV ONLY, 1 pg/mL of TPCK trypsin is added to the medium. 100 pl of virus inoculum is then mixed to 100 pl of each TA concentrations, including virus only wells; 100 pl of medium only is also added to cells only wells for a final 200 pl per well. Plates were incubated at 35°C (for INFV ONLY) or 37°C and 5%CO2 for different number of days (Table 2) before cells fixation and staining.
Immunostaining: For ADV and DENV
[0095] After incubation, media is removed, and cells are fixed with cold 80%/20% (v/v) ethanol/methanol and incubated for 20 minutes at -20oC. Cells are washed 3x with IxDPBS and 200 pL of the diluted anti-virus antibody (Table 4; 1 :2000 in blocking buffer) is added to each well for overnight incubation at 4°C. Primary antibody is removed, and plates are washed three times with IX DPBS. Common secondary antibody, Goat Anti -Mouse IgG (H+L)-HRP Conjugate diluted 1 :2,000 in blocking buffer is then added at 200 pL/well and plates are incubated for 1-hour at room temperature. After removing the secondary antibody, plates are washed three times with IX DPBS. TMB substrate (100 pL/well) is added for approximately 10 minutes (or until virus-only wells are intensely stained while cells-only wells are still weak in signal). Stop solution is then added to each well and optical density is read at 450nm.
Crystal Violet Staining: For HSV-1 and HSV-2
[0096] After incubation, media is removed and cells are fixed and stained with 0.1% crystal violet solution (0.05% methanol, 50% Glutaric dialdehyde in IxDPBS) for 60-minutes +/- 5- minutes at room temperature. Cells are washed 3x with IxDPBS and after removing all liquid, returned plates are air dried at room temperature. Optical density is read at 570nm for each well. Cytotoxicity Assay
[0097] Similar cell and TA preparation is done in parallel for this assay to mimic as close as possible the antiviral assay. Virus inoculum is replaced by serum-free medium only, and for INFV ONLY, 1 pg/mL of TPCK trypsin is added to the medium. To cover all the different incubation times with only 2 conditions, plates are incubated at 35 °C (for INFV ONLY) or 37°C and 5%CO2 for 3 and 5 days. All media is removed and lOOpL of DPBS is added to each to each, following by 100 pL of CellTiter-Glo® reagent. Plates are gently shaken for 2-minutes and incubate the plates at RT for 10-minutes before determining luminescence
Data Evaluation and Data Summary
[0098] All data were imported into Excel. The XLfit 5 plug-in was used with fit# 205 (Levenberg-Marquardt algorithm) to determine the 50 percent inhibition concentration (IC50) and the 50 percent cytotoxicity concentration (CC50). Cytotoxicity (CC50) in Vero cells was determined at 3dpi and 5dpi. Figures 7-12 represents the efficacy data graph. Square points in the graph were manually excluded for better analysis.
Example 14 Inhibition of HCMV infection (Figure 13)
[0099] The antiviral activity of 8 dilutions of compound 1 was explored following administration Ih before infection with HCMV. Compound and virus were left on the cells for the entire duration of the experiment (5 days). The cytotoxicity of the same range of concentrations of compounds was determined by MTT assay. Cells were detached and counted. Count was recorded in the Cell Count Logbook. Cells were seeded in complete media at 4,000 cells/lOOpl/well in four 96 well plates: two for the cytotoxicity assay and two for the infectivity assay. After seeding, the plates were incubated at RT for 5 minutes for even distribution, and then at 37°C, 5% CO2 until the following day. Compound 1 and Acyclovir were diluted to 200 pM in supplemented media, and 225pl of these diluted stocks or diluent only (1% DMSO) were added in triplicate to the top raw (A) of a round bottom 96 well plate, as indicated below.
Pre-treatment of cells 50 pl of supplemented media per well were added to the cells in each plate (infectivity and cytotoxicity). 50 pl per well of treatment from the dilution plate were transferred to the cells in corresponding positions in each plate (infectivity and cytotoxicity). All plates were incubated at 37°C, 5% CO2.
Infection
The virus stock (HCMV Merlin strain, 1x106 lU/ml) was diluted 5-fold with supplemented media to bring the concentration to 2x105 lU/ml. After Ih pre-treatment, media/treatment was removed from the cells and 50 pl per well of treatment from the dilution plate were retransferred to the cells in corresponding positions in the infectivity plates. 50 pl virus per well (MOI ~1) were added, except in column 11 (uninfected control), where 50 pl of supplemented media without virus were added.
Fixation and development
After five days, the infected plates were washed with PBS, fixed for 30 mins with 4% formaldehyde, washed again with PBS, and stored in PBS at 4°C overnight until staining. The cytotoxicity plates were treated with MTT to determine cell viability.
Infectivity readout
Cells were immunostained following commonly known staining protocols, briefly, any residual formaldehyde was quenched with 50 mM ammonium chloride, after which cells were permeabilised (0.1% Triton X100) and stained with an antibody recognising HCMV gB (The Native Antigen Company). The primary antibody was detected with an Alexa-488 conjugate secondary antibody (Life Technologies, A21207), and nuclei were stained with Hoechst. Images were acquired on a Celllnsight CX5 high content confocal microscope (Thermo Fisher) using a 4X objective, and percentage infection calculated using the CX5 software (infected cells/total cells x 100).
Cytotoxicity readout Cytotoxicity was detected by MTT assay following SOP-RA 006. Briefly, the MTT reagent (Sigma, M5655) was added to the cells for 2h at 37°C, 5% CO2, after which the media was removed and the precipitate solubilized with a mixture of 1 : 1 Isopropanol :DMSO for 20 minutes. The supernatant was transferred to a clean plate and signal read at 570nm.
Determintation of TC50 concentration
Percentages of cytotoxicity were calculated using the following formula:
Figure imgf000033_0001
TC50 values were extrapolated from the curves representing the best fit (non-linear regression analysis, variable slope) of the logarithm of compound concentration vs. the normalised percentages of cytotoxicity, using GraphPad Prism (version 9).
Percentages of Cytotoxicity at 5 days post infection
[00100] The graph display the percentage of inhibition of HCMV infection (black line) at different compound concentrations. Sample values in each plate are normalized to the plate internal controls, where 100% inhibition is derived from the average of the negative control (untreated uninfected) and 0% inhibition is derived from the average of the positive control (untreated infected). The x axes show compound dilutions (DM). The curves represent the best fit of the logarithm of compound dilution vs. the normalized percentage of inhibition (variable slope). Cytotoxicity is displayed in gray, with values normalized to the plate internal control (untreated cells, 100% viability).

Claims

WE CLAIM A method of treating a virus infection in a subject in need thereof, comprising administering an effective amount of a diphenyl urea selected but not limited from the group of compounds 1 -7, or a pharmaceutically acceptable salt thereof, to the subject. The method of claim 1, wherein the virus infections are coronavirus causes a SARS or MERS infection, HSV, HCMV, Dengue virus infections and zoonotic pandemic virus infections. The method of claim 1 or 2, wherein the coronavirus causes a SARS-CoV-1 or SARS- CoV-2 or MERS-CoV infection. The method of any one of claims 1-3, wherein the coronavirus is SARS-CoV-2. The method of any one of claims 1-4, wherein the diphenyl urea is 4-{4-[3-(4-Chloro-5- methyl-2-pyrrol-l-yl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide based The method of claim 5, wherein the diphenyl urea is 4-{4-[3-(4-Chloro-5-methyl-2-pyrrol- l-yl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylic acid methylamide, or a pharmaceutically acceptable salt thereof. The method of any one of claims 1-6, wherein the administration of diphenyl urea selected but not limited from the group of compounds 1 -7results in the reduction of the viral load in the subject. The method of claim 7, wherein administration of the diphenyl urea selected but not limited from the group of compounds 1 -7 reduces the viral load by blocking the host pyrimidine synthesis, and thereby blocking the viral replication. The method of claim 7 or 8, wherein the 1 diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to COVID-19 symptoms development. The method of claim 7 or 8, wherein the diphenyl urea selected but not limited from the group of compounds 1 -7 are administered prior to COVID-19 pneumonia development. The method of any one of claims 1-10, wherein the subject has a mild to moderate SARS- CoV-2 infection. The method of any one of claims 1-10, wherein the subject is asymptomatic at the start of the administration regimen. The method of any one of claims 1-12, wherein the subject has had known contact with a patient who has been diagnosed with a SARS-CoV-2 infection. The method of any one of claims 1-13, wherein the subject begins administration of the COVID-19 symptoms development prior to being formally diagnosed with SARS-CoV-2 infection. The method of any one of claims 1-14, wherein the administration of the COVID-19 symptoms development results in one or more clinical benefits. The method of claim 15, wherein the one or more clinical benefits is selected from: shortening the duration of infection, reduction of the likelihood of hospitalization, reduction in the likelihood of mortality, reduction in the likelihood of ICU admission, reduction in the likelihood being placed on mechanical ventilation, reduction in the likelihood supplemental oxygen will be needed, and/or reduction in the length of hospital stay. The method of any one of claims 1-16, wherein the subject is undergoing outpatient treatment. The method of any one of claims 1-17, further comprising administration of one or more additional therapeutic agent. The method of claim 18, wherein the one or more additional therapeutic agents is selected from anti-inflammatories, antibiotics, anti-coagulants, antiparasitic agent, antiplatelet agents and dual antiplatelet therapy, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, beta-blockers, statins and other combination cholesterol lowering agents, specific cytokine inhibitors, complement inhibitors, anti-VEGF treatments, JAK inhibitors, BTK inhibitors, TLR(7/8) inhibitors, immunomodulators, anti- inflammasone therapies, sphingosine-1 phosphate receptors binders, N-methyl-d-aspartate (NDMA) receptor glutamate receptor antagonists, corticosteroids, Granulocytemacrophage colony-stimulating factor (GM-CSF), anti-GM-CSF, interferons, angiotensin receptor-neprilysin inhibitors, calcium channel blockers, vasodilators, diuretics, muscle relaxants, and antiviral medications.
PCT/EP2023/060613 2022-05-16 2023-04-24 Diphenyl ureas for the treatment of viral infections WO2023222332A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP22173527 2022-05-16
EP22173527.7 2022-05-16

Publications (1)

Publication Number Publication Date
WO2023222332A1 true WO2023222332A1 (en) 2023-11-23

Family

ID=81654657

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/060613 WO2023222332A1 (en) 2022-05-16 2023-04-24 Diphenyl ureas for the treatment of viral infections

Country Status (1)

Country Link
WO (1) WO2023222332A1 (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005075425A2 (en) * 2004-01-30 2005-08-18 Merck Patent Gmbh Substituted bisarylurea derivatives as kinase inhibitors
WO2006040056A1 (en) 2004-10-13 2006-04-20 Merck Patent Gmbh Heterocyclic substituted bisarylurea derivatives as kinase inhibitors
US20060241301A1 (en) * 2003-07-30 2006-10-26 Gunter Hoelzemann Urea derivatives and their use as tyrosinkinase inhibitors
WO2007064872A2 (en) * 2005-12-01 2007-06-07 Bayer Healthcare Llc Urea compounds useful in the treatment of cancer
WO2007068383A1 (en) * 2005-12-15 2007-06-21 Bayer Healthcare Ag Diaryl ureas for treating virus infections

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060241301A1 (en) * 2003-07-30 2006-10-26 Gunter Hoelzemann Urea derivatives and their use as tyrosinkinase inhibitors
WO2005075425A2 (en) * 2004-01-30 2005-08-18 Merck Patent Gmbh Substituted bisarylurea derivatives as kinase inhibitors
WO2006040056A1 (en) 2004-10-13 2006-04-20 Merck Patent Gmbh Heterocyclic substituted bisarylurea derivatives as kinase inhibitors
WO2007064872A2 (en) * 2005-12-01 2007-06-07 Bayer Healthcare Llc Urea compounds useful in the treatment of cancer
WO2007068383A1 (en) * 2005-12-15 2007-06-21 Bayer Healthcare Ag Diaryl ureas for treating virus infections

Non-Patent Citations (25)

* Cited by examiner, † Cited by third party
Title
BARNES ET AL., J EXP MED, vol. 217, 2020, pages 6, Retrieved from the Internet <URL:https://doi.org/10.1084/jem.20200652>
CHEN ET AL., LEUKEMIA, vol. 35, 2021, pages 2616 - 2620, Retrieved from the Internet <URL:https://doi.org/10.1038/s41375-021-01266-6>
FUNG ET AL., ANNU. REV. MICROBIOL., vol. 73, 2019, pages 529 - 57
GREIN ET AL., NEJM, 2020
HE, NAT MED, 2020, Retrieved from the Internet <URL:https://doi.org/10.1038/s41591-020-0869-5>
HERSTER ET AL., NAT COMMUN, vol. 11, 2020, pages 105, Retrieved from the Internet <URL:https://doi.org/10.1038/s41467-019-13756-4>
HOFFMANN ET AL., CELL, 2020, Retrieved from the Internet <URL:https:Hdoi.org/10.1016/j.cell.2020.02.052>
KUMAR NIRMAL ET AL: "1,3-Diphenylurea derivatives inhibit cellular entry of influenza A virus and SARS-CoV-2", RESEARCH SQUARE, 26 March 2022 (2022-03-26), XP093062343, Retrieved from the Internet <URL:https://europepmc.org/article/ppr/ppr474869> [retrieved on 20230710], DOI: 10.21203/rs.3.rs-1492489/v1 *
LI ET AL., MICROBES INFECT, vol. 15, no. 2, 2013, pages 88 - 95, Retrieved from the Internet <URL:https://doi.org/10.1016/j.micinf.2012.10.008>
LIMEN ET AL., EXPERT REVIEW OF ANTI-INFECTIVE THERAPY, Retrieved from the Internet <URL:https://doi.org/10.1080/14787210.2021.1982695>
LUDANDEKAR ET AL., NAT. REV. IMM., 2005, Retrieved from the Internet <URL:https://doi.org/10.1038/nri1732>
METHA ET AL., THE LANCET, vol. 395, 2020, pages 10229, Retrieved from the Internet <URL:https://doi.org/10.1016/50140-6736(20)30628-0>
SAITOH ET AL., CELL HOST MICROBE, vol. 19, no. 12(1, 2012, pages 109 - 16
SIDDIQI ET AL., J. HEART AND LUNG TRANS., 2020, Retrieved from the Internet <URL:https://doi.org/10.1016/j.healun.2020.03.012>
STEBBING ET AL., LANCET INFECT DIS, 2020, Retrieved from the Internet <URL:https://doi.org/10.1016/S1473-3099(20)30232-2>
SUNGNAK ET AL.: "Qbio preprint", ARXIV:2003.06122 [Q-BIO.CB
TACCONE ET AL., LANCET RESP. MED., 2020, Retrieved from the Internet <URL:https://doi.org/10.1016/52213-2600(20)30172-7>
YI-CONG ET AL., MINI-REVIEWS IN ORGANIC CHEMISTRY, vol. 16, 2019, pages 7
YIN ET AL., ACS CHEM NEUROSCI, vol. 11, 2020, pages 12, Retrieved from the Internet <URL:https://doi.org/10.1021/acschemneuro.0c00294>
YQI ET AL., BBRC, 2020, Retrieved from the Internet <URL:https://doi.org/10.1016/j.bbrc.2020.03.044>
ZAMBRANA ET AL., SCIENTIFIC REPORTS, vol. 11, 2021, pages 18985, Retrieved from the Internet <URL:https://doi.org/10.1038/s41598-021-98289-x>
ZHAO ET AL., BIORXV PREPRINT, Retrieved from the Internet <URL:https://doi.org/10.1101/2020.01.26.919985>
ZHOU ET AL., LANCET, vol. 395, 2020, pages 1054 - 62
ZOU ET AL., FRONT MED, 2020, Retrieved from the Internet <URL:https://doi.org/10.1007/s11684-020-0754-0>
ZOUQILU ET AL., NAT. REV. IMM., 2018, Retrieved from the Internet <URL:https://doi.org/10.1038/nri.2017.106>

Similar Documents

Publication Publication Date Title
US11491137B2 (en) Methods of improving renal function
US11052073B1 (en) Sphingosine kinase 2 inhibitor for treating coronavirus infection
US20200360347A1 (en) Cenicriviroc for the treatment of fibrosis
US11123329B1 (en) Use of angiotensin II type 2 receptor agonist
WO2022063869A2 (en) Compounds for the treatment of viral infections
US20230129866A1 (en) Methods of treating respiratory disease with deupirfenidone
TW202200139A (en) Tlr7/8 antagonists for the treatment of coronavirus infections
US20230226066A1 (en) Compounds for the treatment of viral infections
WO2023222332A1 (en) Diphenyl ureas for the treatment of viral infections
US20220233480A1 (en) Use of angiotensin ii type 2 receptor agonist
JP2024511141A (en) Treatment of hidradenitis suppurativa with orysmilast
US20230301991A1 (en) Compounds for the treatment of viral infections
US11471448B2 (en) Sphingosine kinase 2 inhibitor for treating coronavirus infection in moderately severe patients with pneumonia
CN116507335A (en) ATR inhibitors for the treatment of viral infections
WO2022058323A1 (en) Compounds for the treatment of viral infections

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23721883

Country of ref document: EP

Kind code of ref document: A1