CYSTEINE PROTEASE INHIBITORS FOR USE IN THE PREVENTION AND/OR TREATMENT OF CORONAVIRUS

Abstract

The present invention relates to a compound according to formula (I) Formula (I) a physiologically acceptable salt, a solvate, or a hydrate thereof for use in the prevention and/or treatment of diseases caused by betacorona-virus infection in a mammalian subject, such as a human, wherein said compound is administered by inhalation as a pharmaceutical composition comprising preferably alcohol as a carrier. A preferred example is viral infection caused by SARS.

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Claims

1. A pharmaceutical composition comprising a compound according to Formula (I), ##STR00006## a physiologically acceptable salt, a solvate, or a hydrate thereof for use in the prevention and/or treatment of diseases caused by betacoronavirus infection in a mammalian subject, wherein said composition is formulated for administration by inhalation and comprises a suitable solvent as a carrier.

2-9. (canceled)

10. A method for preventing and/or treating a disease caused by betacoronavirus infection in a mammalian subject, comprising administering to said subject an effective amount of a compound according to Formula (I), ##STR00007## a physiologically acceptable salt, a solvate, or a hydrate thereof.

11. The method according to claim 10, wherein said method of preventing and/or treating is in combination with another antiviral therapy.

12. The method according to claim 10, wherein said compound is administered to said subject in an effective dosage of about between 1 and 100 mg/kg/day.

13. The method according to claim 10, wherein said compound is administered to said subject at between 5 to 14 days post infection.

14. The method according to claim 10, wherein for prevention said compound is administered to said subject at between 8 to 24 hours post infection.

15. The method according to claim 10, wherein said method further comprises a monitoring step comprising an analysis selected from serum chemistries, liver function tests, hematology, and urinalysis in a sample obtained from said subject, and comparing said analysis to the analysis of an earlier sample from said subject and/or a control sample.

16. The method according to claim 10, wherein said compound is administered by inhalation as a pharmaceutical composition comprising a suitable solvent as a carrier.

17. The method according to claim 16, wherein said suitable solvent is ethanol.

18. The method according to claim 10, wherein said disease is selected from respiratory disease, acute respiratory disease, sepsis, acute respiratory distress syndrome, and adverse immune reactions.

19. The method according to claim 10, wherein said infection is by HCoV-OC43, SARS-CoV-1, HCoV-HKU1, MERS-CoV, SARS-CoV-2 and/or the G614 variant of SARS-CoV-2.

20. The method according to claim 19, wherein said disease is caused by SARS-CoV-2.

21. The method according to claim 11, wherein said another antiviral therapy is selected from remdesivir; interferon alpha 2a or 2b; chloroquine or hydroxychloroquine; serine protease inhibitors; cysteine protease inhibitors; and/or type II transmembrane protease (TMPRSS2) inhibitors.

Description

EXAMPLES

Cytotoxicity of E64D

[0055] E 64 D was found to be non-toxic in a cellular model (Vero e6-cells) in concentrations of at least 100 μM.

Antiviral Effect of E64D-293 ACE2 Cells

[0056] The antiviral effect was measured by CytoPathicEffect (CPE) analysis of SARS-CoV-2 infected 293 ACE2 cells using crystal violet staining for surviving cells. The % of inhibition for the drug as tested was established in duplicate runs. The values were calculated from an OD of 0% inhibition (1% DMSO plus virus) to 100% (1% DMSO without any virus).

[0057] The antiviral activity was found to be at 77.9% and 67.9% inhibition without any substantial cytotoxicity (<4%). In view of this, it is anticipated to use even higher concentrations of more than 100 μM.

Clinical Study on the Effectiveness of E64D in Human Patients

[0058] In this randomized, placebo-controlled trial safety and efficacy of E-64D is assessed in patients hospitalized for severe Covid-19. Patients are included in the study with confirmed SARS-CoV-2 infection who have an oxygen saturation of 94% or less while they were breathing ambient air or who were receiving oxygen support.

[0059] In more detail, patient inclusion parameters are: [0060] Patients who had SARS-CoV-2 infection confirmed by reverse-transcriptase-polymerase-chain-reaction assay [0061] oxygen saturation of 94% or less while the patient was breathing ambient air [0062] a need for oxygen support [0063] patients are required to have a creatinine clearance above 30 ml per minute and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase.sup.1 less than five times the upper limit of the normal range [0064] patients have to agree not to use other investigational agents for Covid-19

[0065] Clinical improvement is assessed in 50 patients receiving a 10-day course of inhaled E-64D three times a day, following 9 days. Follow-up will to continue through at least 28 days after the beginning of treatment with E64D or until discharge or death.

[0066] The primary end point is the time to clinical improvement, defined as the time from randomization to either an improvement of two points on a seven-category ordinal scale or discharge from the hospital. Secondary clinical end point is the reduction of viral load assessed by RT-PCR in the blood.

[0067] The solution formulation of E64D is supplied as a sterile, preservative-free, clear, colorless aqueous-based concentrated solution containing 50 μg/mL E64D. It is supplied as a sterile product in a single-use, clear glass vial with sufficient volume to allow withdrawal of 1.2 mL (60 μg E64D). In addition to the active ingredient, the solution formulation of E64D contains commonly used and pharmaceutically accepted ingredients for inhalation, such as a suitable buffer.

[0068] In the dosage regimen as applied, first, the above amount is inhaled in 6 healthy individuals (healthy volunteers) in order to exclude an adverse irritation of the airways. Lung function is assessed by bodyplethysmography before inhalation and after inhalation of E64D

[0069] Then, for the treatment of established CoV infection, including SARS-CoV, MERS-CoV, and SARS-2-CoV the dosage regimen is as follows: 3 times daily inhalation of 0.4 ml (15 to 30 minute period, total daily volume 1.2 ml) each time until the inhalation chamber is empty (the device has a sensor system that allows the device to shut down after complete inhalation). The recommended E64D dosing duration is a total of 10 days.

[0070] Inhaled doses are administered by an M-neb® dose device and over a 15 to 30 minute period three times a day. After inhalation, no residue remains in the nebulizer unit. The inhaler then turns off.

[0071] During treatment with E64D, the patient is admitted as an inpatient at a facility staffed and maintained by the requesting physician. A peripheral IV line or other venous catheter is maintained. Fluid resuscitation is available if necessary in the event of signs of renal failure or hypotension. Fever is treated with acetaminophen (up to maximum permitted daily dose) and antibiotics as indicated.

[0072] Use of nonsteroidal anti-inflammatory medications and other nephrotoxic agents is avoided, if possible.

[0073] The following laboratory tests are performed daily during therapy: serum chemistries including electrolytes, renal function tests (creatinine, CrCL, BUN), liver function tests (including ALT, AST, total bilirubin, and alkaline phosphatase), hematology (complete blood count and prothrombin time) and urinalysis.

[0074] CoV PCR is performed at regular intervals to monitor response to E64D therapy and to continually weigh the risks and benefits to the patient. Other lab and clinical parameters may be checked at the discretion of the physician.

[0075] Data on patients' oxygen-support requirements, adverse events, and laboratory values, including serum creatinine, ALT, and AST, are reported daily, from day 1 through day 10, and additional follow-up information is solicited through day 28.

[0076] For assessing the results, the incidence of the following key clinical events is quantified: [0077] changes in oxygen-support requirements (ambient air, low-flow oxygen, nasal high-flow oxygen, noninvasive positive pressure ventilation [NIPPV], invasive mechanical ventilation, and extracorporeal membrane oxygenation [ECMO]) [0078] hospital discharge [0079] reported adverse events, including those leading to discontinuation of treatment [0080] serious adverse events [0081] death

[0082] Furthermore, the proportion of patients with clinical improvement is evaluates, as defined by: [0083] live discharge from the hospital [0084] a decrease of at least 2 points from baseline on a modified ordinal scale (as recommended by the WHO R&D Blueprint Group). The six-point scale consists of the following categories: 1, not hospitalized; 2, hospitalized, not requiring supplemental oxygen; 3, hospitalized, requiring supplemental oxygen; 4, hospitalized, requiring nasal high-flow oxygen therapy, noninvasive mechanical ventilation, or both; 5, hospitalized, requiring invasive mechanical ventilation, ECMO, or both; and 6, death. [0085] Virus copies in EDTA blood (before and after treatment)

[0086] E64D is permanently discontinued in the following conditions: [0087] Development of ALT levels>5 times the upper limit of normal [0088] Estimated creatine clearance<30 mL/min based on the Cockcroft-Gault formula