Peptides for preventing or treating covid-19

Abstract

The present invention provides a peptide for use in preventing or treating COVID-19 in a patient, wherein the peptide consists of 7-17 amino acids and comprises the hexamer TX1EX2X3E, wherein X1, X2 and X3 can be any natural or unnatural amino acid, and wherein the peptide does not exhibit TNF-receptor binding activity. Sonapeptide (cyclo-CGQRETPEGAEAKPWYC) is a preferred peptide for use in the present invention.

Description

[0001] The field of this invention relates to peptides for the prevention or treatment of coronavirus disease 2019 (COVID-19).

[0002] COVID-19 is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). See, for example, Rodriguez-Morales et al., 2020, PubMed unique identifier (PMID) 32179124; Jiang et al., 2020, PMID 32133578; and Tay et al., 2020, PMID 32346093. COVID-19 became a pandemic in 2020.

[0003] The most common symptoms of COVID-19 are fever, dry cough, and fatigue. Other less common symptoms include aches and pains, nasal congestion, headache, conjunctivitis, sore throat, diarrhea, loss of taste or smell, or skin rash or discoloration of the fingers or toes.

[0004] Importantly, COVID-19 can affect the lungs, causing pneumonia. In severe cases, COVID-19 can rapidly develop into acute respiratory distress syndrome (ARDS), causing respiratory failure, septic shock, or multiple organ failure. Further complications associated with COVID-19 include sepsis, abnormal blood clotting, and damage to the heart, kidneys, and liver.

[0005] Several drug interventions have been proposed and are currently undergoing clinical trials: these include antiviral drugs such as remdesivir, favipiravir, lopinavir, ritonavir, ribavirin, talivirlin, umefenovir, amantadine, carmostat, and TMC-310911; chloroquines such as hydroxychloroquine and chloroquine; anti-interleukin-6 receptor antibodies such as thalidomide and tocilizumab; interferon-β; and angiotensin-converting enzyme 2. Following results from large clinical trials indicating that remdesivir shortened recovery time, the U.S. Food and Drug Administration (FDA) issued an Emergency Use Authorization (EUA) for the use of remdesivir in hospitalized patients with severe COVID-19 on May 1, 2020. None of these interventions have shown effectiveness in patients with severe and progressive COVID-19.

[0006] In addition, symptomatic treatment is known: patients severely affected by COVID-19 are typically treated in an intensive care unit (ICU) setting with oxygen therapy, mechanical ventilation, renal replacement therapy (RRT), or extracorporeal membrane oxygenation (ECMO). Disease severity is rated from 0 to 8 according to the World Health Organization (WHO), as shown in Table 1 below.

[0007] Table 1 - WHO scoring scheme for COVID-19 severity However, there remains an urgent need for safe and effective pharmaceutical interventions for COVID-19. Therefore, the object of this invention is to provide pharmaceutical prevention or treatment for COVID-19, particularly severe COVID-19 (e.g., severity score higher than 4).

[0008] This invention relates to peptides for the prevention or treatment of COVID-19 in patients, wherein the peptides consist of 7-17 amino acids and include the hexamer TX1EX2X3E (SEQ ID NO: 6), wherein X1, X2, and X3 can be any natural or non-natural amino acids, and wherein the peptides do not exhibit TNF-receptor binding activity. Such peptides are also referred to herein as TIP peptides.

[0009] The present invention also provides a method for delaying the onset of COVID-19 or treating COVID-19, comprising: - To obtain a pharmaceutically acceptable formulation comprising a peptide, wherein the peptide consists of 7-17 amino acids and comprises the hexamer TX1EX2X3E (SEQ ID NO: 6), wherein X1, X2, and X3 may be any natural or non-natural amino acids, and wherein the peptide does not exhibit TNF-receptor binding activity; and - Administer an effective dose of the preparation to patients who have COVID-19 or are at risk of developing COVID-19; Preferably, the peptide is further defined or used as described herein.

[0010] TIP peptide, namely AP301 or sonatide (cyclic-CGQRETPEGAEAKPWYC; CGQRETPEGAEAKPWYC is SEQ ID NO: 1), has been granted compassionate use authorization in Austria and Italy for the treatment of patients with COVID-19. Clinical trials are currently underway (see the Examples section below).

[0011] TIP peptides are peptides containing a human tumor necrosis factor (TNF) lectin-like domain (TIP domain). The TIP domain has been reported, for example, by van der Goot et al., 1999, PMID 10571070. As used herein, TIP peptides consist of 7-17 amino acids comprising the hexamer TX1EX2X3E (SEQ ID NO: 6), wherein X1, X2, and X3 can be any natural or non-natural amino acid, wherein the peptide does not exhibit TNF-specific inflammatory activity (Hribar et al., 1999, PMID 10540321; Elia et al., 2003, PMID 12842853) and can be cyclized. The activity of TIP peptides such as AP301 or sonatide (cyclic-CGQRETPEGAEAKPWYC; CGQRETPEGAEAKPWYC is SEQ ID NO: 1) involves the activation of amiloride-sensitive epithelial sodium channels (ENaC), as reported by Tzotzos et al., 2013, PMID 23313096.

[0012] Unrelated to COVID-19, TIP peptides are known, for example, from European patents EP 1 247 531 B1 and EP 1 264 599 B1 for the treatment of edema, particularly pulmonary edema. Such peptides are also known for the treatment and prevention of vascular complications in diabetic patients, such as microvascular and macrovascular disease, myocardial infarction, microvascular hyperpermeability, stroke, neuropathy, retinopathy, nephropathy, or diabetic foot (EP 2 582 385 B1). Furthermore, such peptides are known for the prevention of edema caused by decreased permeability due to damage to the endothelial and / or epithelial layers (EP 2 403 519 B1, also disclosed in WO 2010 / 099556A1). Additionally, such peptides are known for the treatment and prevention of pulmonary altitude sickness (WO 2014 / 001177 A1). Such peptides are also known to be used for the treatment or prevention of influenza when administered in combination with inhibitors of viral neuraminidase (WO 2012 / 065201 A1). Finally, WO 2015 / 140125 A2 relates to dry powder formulations of TIP peptides.

[0013] The peptides used in this invention are particularly suitable for patients before, after, or during oxygen therapy or ventilation. Thus, in a preferred embodiment, the patient has received or is receiving oxygen therapy or is at risk of receiving oxygen therapy (e.g., via a mask or nasal cannula). In another preferred embodiment, the patient has therefore been or is being ventilated or is at risk of being ventilated, such as by non-invasive ventilation, preferably non-invasive positive pressure ventilation, or by mechanical ventilation.

[0014] In a particularly preferred embodiment, the peptide is used to treat COVID-19 in patients in hospitals (preferably in the ICU) or at risk of being admitted to a hospital (preferably the ICU).

[0015] To further enhance its beneficial effects in the lungs, the peptide is preferably administered to the patient via inhalation (preferably via oral inhalation or intratracheal inhalation).

[0016] The peptide can be administered via a dry powder inhaler (in the pharmaceutical composition). Examples of such powder inhalers that can be used in this invention are described in U.S. Patent Nos. 4,995,385 and 4,069,819; established products include SPINHALER®, ROTAHALER®, FLOWCAPS®, INHALATOR®, DISKHALER®, and AEROLIZER®. Particularly preferred dry powder formulations of the peptide are disclosed in WO 2015 / 140125A2.

[0017] Alternatively, the peptide can be administered via a nebulizer (in the pharmaceutical composition), typically for continuous administration as long as the nebulizer is on or breathing is initiated. Examples of such nebulizers are established products such as Aeroneb® and Pari®. Nebulizers are also disclosed, for example, in document US 9,364,618 B2.

[0018] Finally, the peptide (in the pharmaceutical composition) can be administered via a metered-dose inhaler. A metered-dose inhaler is a device that aerosolizes a predetermined dose of the pharmaceutical composition (i.e., produces a defined dose of a relatively short burst of aerosol) and is typically used for self-administration by a patient. Metered-dose inhalers are disclosed, for example, in US 6,260,549 B1.

[0019] Of course, the peptide can also be administered in other ways, such as intravenously.

[0020] According to a particular preference, the peptide is administered to the patient at a daily dose of 10 mg-500 mg, preferably 50 mg-400 mg, more preferably 75 mg-350 mg, even more preferably 100 mg-300 mg, and even more preferably 125 mg-175 mg, particularly 150 mg-250 mg or even 175 mg-225 mg.

[0021] To further improve efficacy, it is preferable to administer the peptide to the patient for at least two days, preferably at least three days, even more preferably at least four days, and even more preferably at least five days, especially at least six days or even at least seven days.

[0022] This peptide can be administered with other drugs or alone. In a preferred embodiment, the patient has previously experienced, is experiencing, or will experience therapy with at least one compound selected from the following: antiviral drugs such as remdesivir, favipiravir, lopinavir, ritonavir, ribavirin, talivirline, umefenovir, amantadine, carmostat, and TMC-310911; chloroquines such as hydroxychloroquine and chloroquine; anti-interleukin-6 receptor antibodies such as thalidomide and tocilizumab; interferon-β; and angiotensin-converting enzyme 2.

[0023] The peptides disclosed herein are also suitable for prevention, particularly in patients at high risk of SARS-CoV-2 infection, such as healthcare workers treating COVID-19 patients. Therefore, in a preferred embodiment of the invention, the patient is at risk of developing COVID-19, preferably where the patient is a healthcare worker and / or where the patient has been, is, or may have been in close contact with another individual infected with SARS-CoV-2. Administration of the peptide via a dry powder inhaler or a metered-dose inhaler is particularly suitable for such patients. Herein, “close contact” is defined as contact within a 2-meter radius for a minimum duration of 10 minutes.

[0024] In a further preferred embodiment, the peptide of the present invention is used to reduce the risk of a patient developing COVID-19 with a severity score of 5 or 6 according to the WHO scoring scheme (see Table 1 above), preferably wherein the patient has COVID-19 with a severity score of 4 according to the WHO scoring scheme, or to reduce the risk of a patient developing COVID-19 with a severity score of 7 or 8 according to the WHO scoring scheme, preferably wherein the patient has COVID-19 with a severity score of 6 according to the WHO scoring scheme.

[0025] In yet another preferred embodiment, the peptide of the present invention is used to reduce a patient’s COVID-19 severity score according to the WHO scoring scheme (see Table 1 above), specifically from 6 or 7 to 4 or 5.

[0026] The Sequential (or Septic) Organ Failure Assessment (SOFA) score can be used to track a patient's status during their stay in the ICU to determine the extent of organ dysfunction or failure. This score is based on six different scores: one each for the respiratory, cardiovascular, hepatic, coagulation, renal, and nervous systems (see Vincent, JL. et al. "The SOFA (Sepsis-related Organ Failure Assessment) score to describe organ dysfunction / failure." (1996): 707-710., PMID 8844239). A lower SOFA score indicates a lower number of life-threatening organ failures and a higher likelihood of patient survival. This invention demonstrates high suitability for reducing SOFA scores in COVID-19 patients (see Example 2). Therefore, preferably, the (patient to be treated) has an SOFA score of at least 8, preferably at least 9, more preferably at least 10, even more preferably at least 11, and particularly at least 12. In a further preferred embodiment, the invention is used to increase organ function in patients or to reduce SOFA scores in patients.

[0027] Lung compliance is a measure of the lung's ability to stretch and expand (the expandability of elastic tissue). Low compliance indicates stiff lungs, while higher compliance indicates healthier and more elastic lungs. This invention demonstrates a high suitability for increasing lung compliance in COVID-19 patients (see Example 2). Therefore, preferably, the (patient to be treated) has a dynamic lung compliance (also referred to as C) of up to 50, more preferably up to 45, more preferably up to 40, and even more preferably up to 35. dyn (and measured in mL / cm H2O). In a further preferred embodiment, the invention is used to increase patient lung compliance, particularly dynamic lung compliance.

[0028] Normal human body temperature (normothermia, euthermia) is the typical temperature range found in humans. The normal human body temperature range is generally defined as 36.5–37.5°C. Body temperature is a commonly used measure of a patient's overall condition. This invention demonstrates high suitability for lowering body temperature in COVID-19 patients (see Example 2). Therefore, preferably, the patient (to be treated) has a body temperature greater than 37.5°C, particularly greater than 38°C. In a further preferred embodiment, the invention is used to lower the patient's body temperature.

[0029] The invention has also been shown to be particularly suitable for reducing the risk of death (see Example 2). Therefore, in a further preferred embodiment, the invention is used to reduce the risk of death in patients.

[0030] The peptides used in this invention are known in themselves, for example from the following patent documents: EP 1 264 599B1, US 2007 / 299003 A1, WO 94 / 18325 A1, WO 00 / 09149 A1, WO 2006 / 013183 A1 and WO2008 / 148545 A1.

[0031] Preferably, the peptide used according to the present invention comprises the amino acid hexamer TPEGAE (SEQ ID NO: 2), which is a potential carbohydrate-binding motif (Marquardt et al., 2007, PMID 17918767) and can enhance biological activity.

[0032] In a preferred embodiment of the invention, the peptide used according to the invention is cyclic (or ring-shaped) in order to preserve the original TNF-α conformation as much as possible (Elia et al., 2003, PMID 12842853), which may result in higher biological activity. This is an optional feature because, according to Marquard et al., 2007, PMID 17918767, peptide cyclization may not be necessary for carbohydrate binding, which is important for biological activity.

[0033] In a preferred embodiment, the peptide of the present invention comprises the amino acid hexamer TPEGAE (SEQ ID NO: 2). Preferably, the peptide is cyclic and contains a sequential amino acid sequence selected from the following: - QRETPEGAEAKPWY (SEQ ID NO: 3) -PKDTPEGAELKPWY (SEQ ID NO: 4) - CGQRETPEGAEAKPWYC (SEQ ID NO: 1) - CGPKDTPEGAELKPWYC (SEQ ID NO: 5) and - A fragment containing at least seven amino acids of the hexamer TPEGAE (SEQ ID NO: 2). Such peptides or fragments thereof may be biologically active, as illustrated, for example, in WO 2014 / 001177.

[0034] In another preferred embodiment of the invention, the peptide is applied as a single active agent.

[0035] In a particularly preferred embodiment, the peptide according to the invention is peptide AP301 or sonatide (CGQRETPEGAEAKPWYC, SEQ ID NO: 1), wherein the sonatide is cyclized via C residues, preferably via disulfide bonds between C residues.

[0036] In a preferred embodiment, the peptide used in this invention can be formulated in a pharmaceutical composition. The term "pharmaceutical composition" refers to any composition comprising at least one active agent (i.e., a peptide) and preferably one or more excipients, said composition being pharmaceutically acceptable for administration to an individual, particularly a mammal, particularly a human (particularly by inhalation). Suitable excipients are known to those skilled in the art, such as water (particularly water for injection), physiological saline, Ringer's solution, glucose solution, buffers, Hank's solution, vesicle-forming compounds (e.g., lipids), non-volatile oils, ethyl oleate, 5% glucose in saline, substances that enhance isotonicity and chemical stability, buffers, and preservatives such as benzalkonium chloride. The pharmaceutical composition according to the invention can be a liquid or prepared to be dissolved in a liquid such as sterile deionized or distilled water or sterile isotonic phosphate-buffered saline (PBS). Preferably, 1000 µg (dry weight) of this composition comprises or consists of: 0.1-990 µg, preferably 1-900 µg, more preferably 10-200 µg of peptide; and optionally 1-500 µg, preferably 1-100 µg, more preferably 5-15 µg of (buffered) salt (preferably producing isotonic buffer in the final volume); and optionally 0.1-999.9 µg, preferably 100-999.9 µg, more preferably 200-999 µg of other excipients. Preferably, 100 mg of this dry composition is dissolved in sterile deionized / distilled water or sterile isotonic phosphate-buffered saline (PBS) to obtain a final volume of 0.1-100 ml, preferably 0.5-20 ml, more preferably 1-10 ml. However, the dosage and method of administration generally depend on the individual to be treated. Generally, the peptide can be administered at a dose between 1 µg / kg and 10 mg / kg, more preferably between 10 µg / kg and 5 mg / kg, and most preferably between 0.1 and 2 mg / kg.

[0037] As used herein, a peptide according to the invention that is “not TNF-receptor binding activity” or “does not exhibit TNF-receptor binding activity” means that the peptide does not have / does not exhibit TNF-receptor binding activity sufficient to induce TNF-specific inflammatory activity that is detrimental to successful treatment of patients.

[0038] Specifically, “adverse to successful treatment of patients with TNF-specific inflammatory activity” may mean the following: in an in vitro safety pharmacology study of the peptide in human whole blood samples, the study was conducted to assess whether the addition of the peptide resulted in the release of the pro-inflammatory marker interleukin-6 (IL-6) from fresh human whole blood, with the peptide added to the blood sample up to a concentration of 10 mg / ml resulting in the release of less than 0.5 pg / ml of IL-6 (see EP 2 582 385 A1, Example 2).

[0039] In this document, the term "effective dose" (of the peptide) is a dose sufficiently effective to produce the desired therapeutic or preventative effect, such as controlling further deterioration of a disease or condition, treating such deterioration, or promoting cure or eradication of the disease or condition. Typically, the effective dose is formulated for the average patient. However, the actual effective dose (or dosage) may be formulated depending on one or more of the following: a specific administration method; the patient's age, weight, general health status; and the severity and progression of the disease or condition.

[0040] The invention is further described in the following embodiments and figures, but is not limited thereto.

[0041] Figure 1 The mean Sequential Organ Failure Assessment (SOFA) score is shown for COVID-19 patients who were treated with sonatide twice daily over a 7-day period.

[0042] Figure 2 The mean dynamic lung compliance value (in mL / cm H2O) of COVID-19 patients treated twice daily with sonatide over a 7-day period is shown.

[0043] Figure 3 The figure shows the average body temperature (in °C) of COVID-19 patients who were treated twice daily with sonatide over a 7-day period.

[0044] Example 1 - Clinical Trial of COVID-19 Patients On April 11, 2020, the clinical trial of sonatide, “COV-2-SOLNATIDE-20”, began in mechanically ventilated COVID-19 patients (EU Medicines Regulatory Authority Clinical Trials Database - EudraCT No. 2020-001244-26).

[0045] This is a phase II, randomized, placebo-controlled, double-blind trial. Treatment duration for each patient is up to 7 days, and the study duration is up to 28 days. Approximately 40 patients are randomized 1:1 to either the placebo group or the 200 mg daily sonatide group.

[0046] Administration is via intratracheal inhalation through an Aeroneb® nebulizer. Sonatide is formulated as a powder for reconstitution and reconstituted in water for injection.

[0047] Sonatide group: 14 doses of 100 mg sonatide administered to the lungs, each dose reconstituted in water for injection in a double-blind manner, one dose every 12 hours. Placebo group: 14 doses of placebo (i.e., water for injection) administered to the lungs, one dose every 12 hours.

[0048] Endpoints included: number of days without mechanical ventilation (VFD) within 28 days, time to extubation until day 28, all-cause mortality until day 28, changes in scores according to the WHO scoring scheme in Table 1 (assessed on day 14 and day 28 after randomization), number of days in hospital until day 28, and number of days in ICU until day 28.

[0049] It is highly likely that the peptides of the present invention lead to improvements regarding at least one of these endpoints.

[0050] Example 2 Community patients with severe health conditions were taken to local hospitals for examination. Among these patients, positive tests for SARS-CoV-2 via nasopharyngeal swabs and real-time RT-PCR confirmed COVID-19.

[0051] After their condition worsened, critically ill COVID-19 patients were admitted to the hospital's ICU. In addition to standard intensive care and life support, half of the patients received oral sonatide twice daily for seven days.

[0052] In parallel, another cohort of severely ill COVID-19 patients with similar life-threatening conditions were being treated in the hospital's ICU. These patients received only placebo treatment in addition to standard intensive care and life support.

[0053] During the 7-day observation period, patient-centered clinical parameters reflecting the severity of COVID-19 were recorded.

[0054] During the 7-day observation period, patient-centered clinical parameters reflecting the severity of COVID-19 were recorded.

[0055] The SOFA score is used to track a patient's condition during their stay in the intensive care unit (ICU) to determine the extent of organ dysfunction or failure.

[0056] Figure 1 The average SOFA score is shown for COVID-19 patients treated twice daily with sonatide over a 7-day period. During treatment, the average SOFA score decreased from 12 to 8.

[0057] In contrast, the SOFA score of severe COVID-19 patients who were not treated with sonatide did not decrease over the same interval.

[0058] Figure 2 The values ​​show the mean dynamic lung compliance of COVID-19 patients treated twice daily with sonatide over a 7-day period. During treatment, mean dynamic lung compliance increased from approximately 35 mL / cm H2O to 50 mL / cm H2O.

[0059] In contrast, dynamic lung compliance in severe COVID-19 patients who were not treated with sonatide remained below 40 mL / cm H2O at the same intervals.

[0060] Figure 3 This shows the average body temperature of COVID-19 patients treated twice daily with sonatide over a 7-day period. The average body temperature increased at the start of treatment but decreased during the 7-day treatment period.

[0061] In contrast, severe COVID-19 patients who were not treated with sonatide consistently had body temperatures above 38°C.

[0062] Scientific literature indicates that the mortality rate for severe COVID-19 patients is as high as 50% among those treated in the ICU (Tzotzos et al. Incidence of ARDS and outcomes in hospitalized patients with COVID-19. Critical Care (2020) 24:516; PMID 32825837). In this study, none of the severe COVID-19 patients treated with sonatide died within 28 days of the onset of COVID-19 symptoms. In contrast, in a comparative cohort consisting of patients with similarly severe COVID-19, three patients died within 28 days. sequence list <110> Apeptico Research and Development LLC <120> Peptides used to prevent or treat COVID-19 <130> R77134 <150> 20173713.7 <151> 2020-05-08 <160> 6 <170> BiSSAP 1.3 <210> 1 <211> 17 <212> PRT <213> Artificial sequence <220> <223> AP301 peptide <400> 1 Cys Gly Gln Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr 1 5 10 15 Cys <210> 2 <211> 6 <212> PRT <213> Artificial sequence <220> <223> peptide sequence <400> 2 Thr Pro Glu Gly Ala Glu 1 5 <210> 3 <211> 14 <212> PRT <213> Artificial sequence <220> <223> peptides <400> 3 Gln Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr 1 5 10 <210> 4 <211> 14 <212> PRT <213> Artificial sequence <220> <223> peptides <400> 4 Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr 1 5 10 <210> 5 <211> 17 <212> PRT <213> Artificial sequence <220> <223> peptides <400> 5 Cys Gly Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr 1 5 10 15 Cys <210> 6 <211> 6 <212> PRT <213> Artificial sequence <220> <223> peptide sequence <220> <221> variants <222> 2,4,5 <223> Any natural or non-natural amino acid <400> 6 Thr Xaa Glu Xaa Xaa Glu 1 5

Claims

1. Use of the peptide in the preparation of a medicament for the prevention or treatment of COVID-19 in patients, wherein the peptide is CGQRETPEGAEAKPWYC (SEQ ID NO: 1) and is cyclized via disulfide bonds between C residues.

2. The use according to claim 1, wherein the peptide is administered to the patient by inhalation.

3. The use according to claim 2, wherein the inhalation is oral inhalation or intratracheal inhalation.

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