WO2022040803A1

WO2022040803A1 - Use of c-terminal soricidin peptides for the treatment or prevention of sars-cov2 infection

Info

Publication number: WO2022040803A1
Application number: PCT/CA2021/051186
Authority: WO
Inventors: John M. Stewart; Dominique Dugourd
Original assignee: Soricimed Biopharma Inc.
Priority date: 2020-08-26
Filing date: 2021-08-26
Publication date: 2022-03-03

Abstract

The present disclosure relates to methods and uses of C-terminal Soricidin peptides for inhibiting SARS CoV-2 S-protein interactions with Angiotensin Converting Enzyme-2 (ACE2), inhibiting SARS CoV-2 binding and entry into cells, and/or treating or preventing SARS CoV-2 infection.

Description

USE OF C-TERMINAL SORICIDIN PEPTIDES FOR THE TREATMENT OR PREVENTION OF SARS-COV2 INFECTION

CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63/070,525, filed August 26, 2020, the contents of which are incorporated herein by reference in its entirety.

SEQUENCE LISTING

[0002] A computer readable form of the Sequence Listing “15309- P62396PC00_SequenceListing” (1 ,835 bytes) created on August 25, 2021 , is herein incorporated by reference.

FIELD

[0003] The disclosure relates to the treatment or prevention of infections with SARS-CoV2 and more specifically to the use of C-terminal soricidin peptides for the treatment or prevention of infections with SARS CoV-2 and coronavirus disease (COVID- 19).

INTRODUCTION

[0004] SARS CoV-2 attaches to, and is internalized through the interaction of its Spike (S) protein with Angiotensin Converting Enzyme 2 (ACE2) on the cell surface. The RBD (receptor binding domain) of SARS CoV-2 is at the tip of the extended (or open) conformation of the Spike proteins. There are three identical subunits that form the end of Spike assembly and any, or all, can be folded outward from the virus surface to interact with ACE2. Inhibiting the interaction between the RBD of the viral SARS CoV-2 Spike protein and ACE2 may prevent entrance of SARS CoV-2 into the cell, thereby curtailing or preventing infection and any resulting disease such as COVID-19.

[0005] SARS CoV and SARS CoV-2 infections can cause a hyper-inflammatory or cytokine storm response from the immune system. Virally infected cells initiate formation of a calcium-dependent inflammasome. SARS CoV and SARS CoV-2 produce an envelope protein (E) that can behave as a calcium channel. E protein increases cytoplasmic calcium, activating the inflammasome via Nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain-containing protein 3 (NLRP3) (Nieto-Torres et al., 2015). Activation of the inflammasome results in the production of I L-1 p and triggers an inflammatory response. [0006] Early results indicate that suppression of the inflammatory response is beneficial for treating COVID-19. Treatment with an interleukin-1 receptor antagonist (anakinra) resulted in clinical improvement of 72% of trial subjects (Cavalli et al., 2020). Tocilizumab, an antibody against IL-6 receptor, reduced death and ICU admissions to 25% of patients from 72% of patients who did not receive the anti-IL-6R treatment (Klopfenstein et al., 2020).

[0007] Soricidin (NCBI accession no. P0C2P6) is a fifty-four amino acid paralytic peptide isolated from the submaxilary saliva gland of the Northern Short-tailed Shrew (Blarina brevicauda). Previous patents have described isolation of the soricidin peptide and provided data showing that the 54-mer peptide caused paralysis and inhibited calcium uptake in two ovarian cancer cell lines (see US patent nos. 7,119,168 and 7,273,850).

[0008] Peptides corresponding to certain C-terminal sequences of soricidin have been shown to inhibit Transient Receptor Potential Vanilloid channel 6 (TRPV6) without paralytic activity and to be useful for the treatment of cancer, including metastatic cancer (see e.g. W02009114943). The peptides maintain TRPV6 calcium channel binding activity without the paralytic activity of the full-length soricidin peptide. C-terminal soricidin peptides have also been shown to activate TRPV3 and be useful for promoting skin repair (PCT patent publication no. WO2014040178)

SUMMARY

[0009] C-terminal fragments of soricidin are shown herein to inhibit the interaction between the SARS CoV-2 Spike protein and ACE2. As shown in Example 1 , computer modeling of SOR-C13 with SARS CoV-2 Spike protein suggested an unexpected binding of the peptide to a region of the open conformation of S protein that interacts with ACE2. Further testing using an in vitro immunoassay demonstrated that SOR-C13 inhibits the interaction of SARS-CoV-2 spike protein with human ACE2. Remarkably, smaller fragments of SOR-C13 including SOR-C9, SOR-C6 and SOR-C5 were also demonstrated to inhibit the interaction between SARS CoV-2 spike protein with human ACE2. Accordingly, the peptides described herein are expected to be useful for the treatment or prevention of and COVID-19.

[0010] SOR-C13 is known to inhibit the TRPV6 calcium channel and decrease cellular calcium concentrations. Reduced cellular calcium is expected to reduce the induction or activity of NLRP3 and/or the inflammasome in SARS CoV-2 infected cells, resulting in lowered production of IL-1 p and reduction of the inflammatory response and/or cytokine storm. Accordingly, the peptides described herein are also expected to be useful for the prevention or treatment of SARS CoV-2-induced hyper-inflammatory response.

[0011] SARS CoV-2 infections can cause a hyper-inflammatory or cytokine storm response from the immune system where infected cells initiate formation of a calciumdependent inflammasome. As set out in Example 3, use of the peptides described herein may also reduce cytosolic calcium levels and/or dampen inflammasome activity following SARS CoV-2 infection and thereby avoid the undesirable cytokine storm response seen in some patients.

[0012] Accordingly, in one aspect there is provided a method of treating or preventing SARS CoV-2 infection in a subject in need thereof, the method comprising administering to the subject an effective amount of a peptide comprising all or part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1).

[0013] Also provided is the use of an effective amount of a peptide comprising all or part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1) for treating or preventing SARS CoV-2 infection in a subject in need thereof. Also provided is the use of an effective amount of a peptide comprising all or part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1 ) in the manufacture of a medicament for treating or preventing SARS CoV-2 infection in a subject in need thereof.

[0014] In an embodiment, the subject has or is suspected of having SARS CoV-2. In an embodiment the subject has been diagnosed with a SARS CoV-2 infection, optionally using a diagnostic test such as a nucleic acid amplification test (NAAT), optionally a polymerase chain reaction (PCR) test.

[0015] In an embodiment, the peptide inhibits binding of SARS CoV-2 spike protein to human angiotensin converting enzyme 2 (ACE2).

[0016] In an embodiment, the peptide binds to the SARS CoV-2 spike protein, optionally the ACE2 receptor-binding domain of the SARS CoV-2 spike protein.

[0017] In an embodiment, the peptide reduces or suppresses an inflammatory response to SARS CoV-2 infection in the subject.

[0018] In an embodiment, the peptide inhibits Transient Receptor Potential Vanilloid subfamily member s (TRPV6) calcium channel activity. [0019] In an embodiment, the peptide reduces cytoplasmic calcium influx and/or reduces activation or activity of the inflammasome.

[0020] In an embodiment, the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ I D NO: 1 .

[0021] In an embodiment, the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ I D NO: 1 .

[0022] In an embodiment, the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1), optionally wherein the peptide is in the form of an acetate salt.

[0023] In an embodiment, the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

[0024] In an embodiment, the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

[0025] In one embodiment, the methods and uses described herein include the use or administration of a nucleic acid molecule encoding a peptide encoding a C-terminal soricidin peptide that inhibits the interaction between and ACE2 to a subject in need thereof. For example, in one embodiment, an expression vector comprising a nucleic acid molecule encoding a C-terminal soricidin peptide described herein operably linked to a promoter is used or administered to a subject for treating or preventing SARS CoV-2 infection in a subject in need thereof.

[0026] In an aspect, there is provided a method for inhibiting binding of SARS CoV- 2 spike protein to a cell expressing human angiotensin converting enzyme 2 (ACE2), the method comprising exposing the cell to a peptide comprising all of part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1).

[0027] In an embodiment, the peptide binds to the SARS CoV-2 spike protein, optionally the ACE2 receptor-binding domain of the SARS CoV-2 spike protein.

[0028] In an embodiment, the cell is in vivo, in vitro or ex vivo. [0029] In an embodiment, the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ I D NO: 1 .

[0030] In an embodiment, the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ I D NO: 1 .

[0031] In an embodiment, the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1), optionally wherein the peptide is in the form of an acetate salt.

[0032] In an embodiment, the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

[0033] In an embodiment, the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

[0034] Other features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating preferred embodiments, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from the detailed description.

DRAWINGS

[0035] Embodiments of the disclosure will be described in relation to the drawings in which:

[0036] Figure 1 shows the closed (LEFT; 6vxx.pdb) and open (RIGHT; 6vyb.pdb) conformations of one of the subunits of the SARS CoV-2 Spike protein. The white arrow on the right image indicates the tip of the Spike protein, folded open, which interacts with ACE2 (Walls et al., 2020).

[0037] Figure 2 shows the interaction of the 50 best SOR-C13 structures with the open (LEFT) and closed (RIGHT) conformations of SARS CoV-2 Spike protein. [0038] Figure 3 shows a close-up of the ACE2/Spike interface region with and without SOR-C13. The left hand image shows the edge of the extended Spike protein that interacts with ACE2. The right hand image shows the first-ranked conformation of SOR-C13 in a binding cavity uncovered by extension of the Spike protein.

[0039] Figure 4 shows: (TOP) the interaction propensity at each amino acid position of SARS CoV-2 Spike protein with B chain in open conformation; A and C chains are in closed conformation. The top black line indicates the region at the tip of an opened S-protein subunit with the shorter line indicating the amino acids at the interface with ACE2. (BOTTOM) the Propensity value at each amino acid position for SARS CoV-2 Spike protein in closed conformation shows only weak and non-specific interaction.

[0040] Figure 5 shows a comparison of the docking of SOR-C13 to the open conformation of the Spike protein of SARS CoV-2 (LEFT) and SARS CoV-1 (RIGHT). The Receptor Binding Domain of both images is indicated by an arrow.

[0041] Figure 6 shows a comparison of the interaction of a series of peptides with the open conformation of SARS CoV-2 Spike protein. The region of the RBD that interacts with ACE2 is in the 490 to 500 amino acid region indicated in the graphs by the heavy bar over this region.

[0042] Figure 7 shows the dependence of the docking propensity of Spike open B chain on length of the docking peptide sequence.

[0043] Figure 8 shows a comparison of the relative number of 200 peptide configurations that interact with the RBD of SARS CoV-2 open Spike protein. SOR-C27 appears to bind to the outside and the inside of the extended Spike RBD. The lower data point is the internal RBD face, while the upper data point is the total number.

[0044] Figure 9 shows a three-dose rapid test of inhibition of the interaction between SARS CoV-2 Spike protein. An antibody to the Spike protein was used as the positive control (at 0 10 nM and 100 nM). An assortment of peptides were tested at 0, 10 uM and 100 uM. SOR-C13A = acetate salt of SOR-C13; SOR-C13 N+A = amidated/acetylated SOR-C13. SOR-C13T = trifluoroacetate (TFA) salt of SOR-C13. All other peptide were as the TFA salt.

[0045] Figure 10 shows a dose-response curve of inhibition of the interaction between SARS CoV-2 Spike protein. An antibody to the Spike protein was used as the positive control. The peptide is acetate salt of SOR-C13. The concentrations tested were 0.01 , 0.05, 0.10, 0.50, 1.00, 5.00, 10.0, 50.0 and 100.0 nM (for antibody) oruM forpeptide.

DESCRIPTION OF VARIOUS EMBODIMENTS

[0046] The inventors have shown that C-terminal Soricidin peptides are useful for the prevention and/or treatment of SARS CoV-2 viral infection. In particular, C-terminal Soricidin peptides have been shown to bind the spike (S) protein of SARS CoV-2 and thereby inhibit the interaction of spike protein with the cell surface protein Angiotensinconverting enzyme 2 (ACE2). C-terminal Soricidin peptides are also known to inhibit Transient Receptor Potential Vanilloid subfamily member 6 (TRPV6) and reduce cytoplasmic calcium influx. As set out in Example 3, C-terminal Soricidin peptides are therefore also expected to reduce activation or activity of the inflammasome and may help avoid or suppress the undesirable cytokine storm response seen in some patients infected with SARS CoV-2.

[0047] As used herein, “SARS CoV-2” refers to Severe Acute Respiratory Syndrome coronavirus-2, which is the causative agent of Coronavirus disease 2019 (COVID-19).

[0048] As used herein, “spike protein” or “S protein” refer to the spike protein of SARS CoV-2 such as those described in RCSB PDS accession no. 6VYB and NCBI accession nos.6VYB_A, 6VYB_B and 6VYBC.

[0049] As used herein, “C-terminal Soricidin peptides” or “Soricidin peptides” refer to peptides having all or part of the sequence KEFLHPSKVDLPR (SEQ ID NO: 1) and variants, conjugates, and salts thereof. In one embodiment, the C-terminal soricidin peptides inhibit the interaction between the SARS CoV-2 spike protein and ACE2.

[0050] In one embodiment, the peptides described herein comprise or consist of all or part of SOR-C13 (KEFLHPSKVDLPR; SEQ ID NO: 1) and inhibit the interaction between the SARS CoV-2 spike protein and ACE2. In one embodiment, the peptide comprises or consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1.

[0051] In an embodiment, the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1 ), HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), or VDLPR (SOR-C5; SEQ ID NO: 5). In an embodiment, the peptide comprises or consists of an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to a sequence identified herein as inhibiting the interaction between and ACE2 such as KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1 ), HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5)

[0052] The peptides described herein optionally also include analogs of the aforementioned peptides. Analogs of the peptides optionally include, but are not limited to an amino acid sequence containing one or more amino acid substitutions, insertions, deletions and/or mutations. Amino acid substitutions may be of a conserved or nonconserved nature. Conserved amino acid substitutions involve replacing one or more amino acids of the peptide with amino acids of similar charge, size, and/or hydrophobicity characteristics. When only conserved substitutions are made, the resulting analog should be functionally equivalent. Non-conserved substitutions involve replacing one or more amino acids of the amino acid sequence with one or more amino acids, which possess dissimilar charge, size, and/or hydrophobicity characteristics. The analog is optionally a peptoid, which is an N-substituted polyglycine with amino acid R groups attached at the N atom.

[0053] One or more amino acid insertions or deletions are optionally introduced into the peptide sequences described herein. Amino acid insertions consist of single amino acid residues or sequential amino acids ranging for example from 2 to 15 amino acids in length.

[0054] Deletions consist of the removal of one or more amino acids, or discrete portions from the amino acid sequence of the peptide. The deleted amino acids may or may not be contiguous.

[0055] The peptides described herein are readily prepared by chemical synthesis using techniques well known in the chemistry of proteins such as solid phase synthesis (Merrifield, 1964, J. Am. Chem. Assoc. 85:2149-2154) or synthesis in homogenous solution (Houbenweyl, 1987, Methods of Organic Chemistry, ed. E. Wansch, Vol. 15 1 and II, Thieme, Stuttgart). The peptides also include peptides having sequence identity to a C-terminal Soricidin peptide, mutated peptides and/or truncations thereof as described herein. Alternatively, the peptides may be prepared using recombinant protein expression systems. [0056] Analogs of a peptide described herein are optionally prepared by introducing mutations in a nucleotide sequence encoding the peptide. Mutations in nucleotide sequences constructed for expression of analogs of a peptide preserve the reading frame of the coding sequences. Furthermore, the mutations will preferably not create complementary regions that could hybridize to produce secondary mRNA structures such as loops or hairpins, which could adversely affect translation of the mRNA.

[0057] Mutations are optionally introduced at particular loci by synthesizing oligonucleotides containing a mutant sequence, flanked by restriction sites enabling ligation to fragments of the native sequence. Following ligation, the resulting reconstructed sequence encodes an analog having the desired amino acid insertion, substitution, or deletion.

[0058] Alternatively, oligonucleotide-directed site-specific mutagenesis procedures are employed to provide an altered gene having particular codons altered according to the substitution, deletion, or insertion required. Deletion or truncation of a peptide of the invention is also readily achieved by utilizing convenient restriction endonuclease sites adjacent to the desired deletion. Subsequent to restriction, overhangs may be filled in, and the DNA re-ligated. Exemplary methods of making the alterations set forth above are disclosed by Sambrook et al (Sambrook J et al. 2000. Molecular Cloning: A Laboratory Manual (Third Edition), Cold Spring Harbor Laboratory Press).

[0059] Other useful peptides optionally comprise, consist essentially of or consist of an amino acid sequence with at least: 50%, 60%, 70%, 80%, 90% or 95% sequence identity to all or part of SEQ ID NO:1 described herein, wherein the peptide has SARS CoV-2 spike protein inhibition activity, has TRPV6 inhibition activity, and/or is useful for treatment or prevention of SARS CoV-2. Sequence identity is typically assessed by the BLAST version 2.1 program advanced search (parameters as above; Altschul, S.F., Gish, W., Miller, W., Myers, E.W. & Lipman, D.J. (1990) "Basic local alignment search tool." J. Mol. Biol. 215:403_410). BLAST is a series of programs that are available online through the U.S. National Center for Biotechnology Information (National Library of Medicine

Building 38A Bethesda, MD 20894) The advanced Blast search is set to default parameters. References for the Blast Programs include: Altschul, S.F., Gish, W., Miller, W., Myers, E.W. & Lipman, D.J. (1990) "Basic local alignment search tool." J. Mol. Biol. 215:403-410; Gish, W. & States, D.J. (1993) "Identification of protein coding regions by database similarity search." Nature Genet. 3:266-272.; Madden, T.L., Tatusov, R.L. & Zhang, J. (1996) "Applications of network BLAST server" Meth. Enzymol. 266:131-141 ; Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D.J. (1997) "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs." Nucleic Acids Res. 25:3389-3402); Zhang, J. & Madden, T.L. (1997) "PowerBLAST: A new network BLAST application for interactive or automated sequence analysis and annotation." Genome Res. 7:649-656).

[0060] The peptides described herein also includes a peptide conjugated with a selected protein, or a selectable marker protein to produce fusion proteins.

[0061] In an embodiment, the peptide is SOR-C13 having the sequence KEFLHPSKVDLPR (SEQ ID NO: 1 ) or has a sequence with at least: 50%, 60%, 70%, 80%, 90% or 95% sequence identity with SEQ ID NO: 1. In one embodiment, the peptide has at least or about 25%, 50%, 70%, 80% or at least or about 90% SARS CoV-2 S protein inhibition activity compared with a peptide having the sequence of SEQ ID NO: 1 .

[0062] In another embodiment, the peptide is SOR-C9 having the sequence HPSKVDLPR (SEQ ID NO: 3), or has a sequence with at least: 50%, 60%, 70%, 80%, 90% or 95% sequence identity with SEQ ID NO: 3. In one embodiment the peptide has at least or about 25%, 50%, 70%, 80% or at least or about 90% SARS CoV-2 S protein inhibition activity compared with peptides having the sequence of SEQ ID NO: 3.

[0063] In another embodiment, the peptide is SOR-C6 having the sequence KVDLPR (SEQ ID NO: 4), or has a sequence with at least: 50%, 60%, 70%, 80%, 90% or 95% sequence identity with SEQ ID NO: 4. In one embodiment the peptide has at least or about 25%, 50%, 70%, 80% or at least or about 90% SARS CoV-2 S protein inhibition activity compared with peptides having the sequence of SEQ ID NO: 4.

[0064] In another embodiment, the peptide is SOR-C5 having the sequence VDLPR (SEQ ID NO: 5), or has a sequence with at least: 50%, 60%, 70%, 80%, 90% or 95% sequence identity with SEQ ID NO: 5. In one embodiment the peptide has at least or about 25%, 50%, 70%, 80% or at least or about 90% SARS CoV-2 S protein inhibition activity compared with peptides having the sequence of SEQ ID NO: 5.

[0065] In an embodiment, the peptide is an acetate salt or a trifluoroacetate (TFA) salt. In an embodiment, the peptide is amidated and/or acetylated. [0066] Peptides having SARS CoV-2 spike protein inhibition activity are readily identified using any known assays suitable for measuring protein binding and/or viral entry into cells. For example, the protein binding immunoassays described in Example 2 may be used to test peptides for inhibiting the interaction between SARS CoV-2 spike protein and ACE2.

[0067] In an embodiment, the peptide inhibits spike protein/ACE2 interaction with an ICso of about 4.5 uM or less. In one embodiment, the peptide inhibits spike protein/ACE2 interaction with an IC50 of less than or equal to about 50 uM, 25 uM, 10 uM, 5 uM, or 4.5 uM.

[0068] In one embodiment, the disclosure includes methods of reducing the interaction of SARS CoV-2 spike protein with a cell expressing the cell surface protein ACE2 by administering a peptide described herein to the cell or to a sample suspected of containing SARS CoV-2, wherein the peptide binds to the SARS CoV-2 spike protein and inhibits its interaction with a cell expressing ACE2. Reduction in interaction can be determined using any suitable assay for example flow cytometry or fluorescence microscopy techniques, or by an immunoassay such as the protein binding immunoassay described in Example 2.

[0069] In one embodiment, the peptides described herein inhibit the activity of a calcium channel, TRPV6 (Bowen et al., 2013), reducing cellular calcium levels. Reduced cellular calcium may reduce the activation or activity of NLRP3 and/or the inflammasome, resulting in lowered production of IL-1 p and reduction of the inflammatory response. Accordingly, in an embodiment, exposing cells to the peptides described herein such as by administration to a subject results in reduced activation or activity of NLRP3 and/or the inflammasome, lowered production of IL-1 p, and/or reduction of the inflammatory response in response to infection with SARS CoV-2. In another embodiment, the peptides described herein are useful for the prevention or treatment of SARS CoV-2-induced NLRP3 and/or inflammasome activation or activity, and/or SARS CoV-2-induced inflammatory response in a subject.

[0070] The activation or activity of NLRP3 and/or the inflammasome, production of IL-1 p, and/or the inflammatory response in response to SARS CoV-2 can be measured or detected using any suitable method. For example, expression levels of IL-1 p and/or other components of the inflammatory response can be measured by real time PCR, ELISA, or any other suitable method.

[0071] In one embodiment, an effective amount of a peptide comprising all or part of SEQ ID NO:1 as described herein, is useful for inhibiting SARS CoV-2 S protein/ACE2 interaction, reducing SARS CoV-2 viral entry into cells, and/or preventing or treating SARS CoV-2 infection by administration of the peptide and/or its use in a subject in need thereof.

[0072] The term “administration” or "administered" as used herein means administration of a therapeutically effective amount of a peptide of the disclosure to a cell or subject. The peptides described herein can be administered for example, by topical, parenteral, intravenous, subcutaneous, intramuscular, intracranial, intraventricular, intrathecal, intraorbital, ophthalmic, intraspinal, intracisternal, intraperitoneal, intranasal, aerosol or oral administration. In certain embodiments, the pharmaceutical composition is administered topically. Administering a peptide or substance to a subject includes both in vivo and ex vivo administrations.

[0073] As used herein, the phrase "effective amount" or "therapeutically effective amount" means an amount effective, at dosages and for periods of time necessary to achieve the desired result (e.g. optionally inhibiting SARS CoV-2 S protein/ACE2 interaction, reducing SARS CoV-2 entry into cells, and/or preventing or treating SARS CoV-2 infection). For example, in the context of treating a viral infection, an effective amount is an amount that achieves a treatment response, for example reduces disease burden, and/or reduces or prevents proliferation or spreading of viral particles, as compared to the response obtained without administration of the compound. The amount of a given compound that will correspond to such an amount will vary depending upon various factors, such as the given peptide, the pharmaceutical formulation, the route of administration, the type of disease or disorder (for example the strain of SARS CoV-2), the type of disease presentation, the identity of the subject or host being treated, and the like, but can nevertheless be routinely determined by one skilled in the art. Effective amounts may vary for example according to the type, location, or extent of the SARS CoV-2 infection, or factors such as the age, sex and weight of the subject.

[0074] The term "subject" as used herein includes all members of the animal kingdom including mammals, and suitably refers to humans. Optionally, the term “subject” includes mammals that have been diagnosed with a SARS CoV-2 infection. In one embodiment, the term “subject” refers to a human having, or suspected of having, a SARS CoV-2 infection. In an embodiment, the subject has been diagnosed as having a SARS CoV-2 infection, optionally using a diagnostic test such as a molecular diagnostic test or an antigen test.

[0075] As used herein, “diagnostic test” means a test which detects an active infection. Diagnostic tests can detect the presence of viral nucleic acids or proteins (antigens), and include for example nucleic acid amplification tests (NAATs) such as polymerase chain reaction (PCR) tests, optionally a reverse transcription PCR (RT-PCR), or transcription mediated amplification (TMA) assays.

[0076] The term "a cell" as used herein includes a single cell as well as a plurality or population of cells. Administering a peptide or substance to a cell includes both in vitro and in vivo administrations.

[0077] The peptides described herein, by inhibiting the SARS CoV-2 S protein/ACE2 interaction, reduce SARS CoV-2 entry into cells and/or prevent or treat SARS CoV-2 infection when administered to a subject such as a human subject. Therefore, an embodiment includes the use of a peptide described herein for reducing SARS CoV-2 viral entry into cells and/or preventing or treating SARS CoV-2 infection.

[0078] As used herein, and as well understood in the art, "to treat" or "treatment" is an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease or disorder, preventing spread of disease or disorder, delay or slowing of disease or disorder progression, amelioration or palliation of the disease or disorder state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0079] The terms “prevent”, “preventing”, or “prevention” as used herein and as understood in the art, means an approach for blocking or halting the development of a condition or disease state such as a viral infection. “Preventing” and “prevention” as used herein also include prophylactic use or administration. [0080] The invention also includes the use of the peptides described herein for preparation of a medicament for treatment of infection caused by SARS CoV-2. The isolated peptides described herein are optionally formulated into a pharmaceutical composition for administration to subjects in a biologically compatible form suitable for administration in vivo. By "biologically compatible form suitable for administration in vivo" is meant a form of the substance to be administered in which any toxic effects are outweighed by the therapeutic effects. The substances may be administered to living organisms including humans, and animals.

[0081] Administration of a therapeutically active amount of pharmaceutical compositions of the present invention is defined as an amount effective, at dosages and for periods of time necessary to achieve the desired result. For example, a therapeutically active amount of a substance may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance to elicit a desired response in the individual. Dosage regimes may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.

[0082] The peptide described herein is preferably combined with other components such as a pharmaceutically acceptable carrier in a composition such as a pharmaceutical composition. The compositions are useful when administered in methods of medical treatment or prevention of SARS CoV-2 infection.

[0083] The pharmaceutical compositions can be administered to humans or animals by a variety of methods including, but not restricted to topical administration, oral administration, aerosol administration, intratracheal instillation, intraperitoneal injection, injection into the cerebrospinal fluid, intravenous injection and subcutaneous injection. Dosages to be administered depend on patient needs, on the desired effect and on the chosen route of administration. Nucleic acid molecules and peptides may be introduced into cells using in vivo delivery vehicles such as liposomes. They may also be introduced into these cells using physical techniques such as microinjection and electroporation or chemical methods such as co-preci pitation, pegylation or using liposomes.

[0084] The pharmaceutical compositions are prepared by known methods for the preparation of pharmaceutically acceptable compositions which can be administered to patients, and such that an effective quantity of the nucleic acid molecule or peptide is combined in a mixture with a pharmaceutically acceptable vehicle. Suitable vehicles are described, for example in Remington's Pharmaceutical Sciences (Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., USA) or Handbook of Pharmaceutical Additives (compiled by Michael and Irene Ash, Gower Publishing Limited, Aidershot, England (1995). On this basis, the compositions include, albeit not exclusively, solutions of the substances in association with one or more pharmaceutically acceptable vehicles or diluents, and may be contained in buffered solutions with a suitable pH and/or be iso-osmotic with physiological fluids. In this regard, reference can be made to U.S. Pat. No. 5,843,456.

[0085] On this basis, the pharmaceutical compositions optionally includes an active compound or substance, such as a peptide or nucleic acid molecule, in association with one or more pharmaceutically acceptable vehicles or diluents, and contained in buffered solutions with a suitable pH and iso-osmotic with the physiological fluids. The methods of combining the active molecules with the vehicles or combining them with diluents are well known to those skilled in the art. The composition optionally includes a targeting agent for the transport of the active compound to specified sites within tissue.

[0086] The above disclosure generally describes the present application. A more complete understanding can be obtained by reference to the following specific examples. These examples are described solely for the purpose of illustration and are not intended to limit the scope of the disclosure. Changes in form and substitution of equivalents are contemplated as circumstances might suggest or render expedient. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.

[0087] All publications and references to biological sequences contained herein are hereby incorporated by reference in their entirety.

EXAMPLES

[0088] The following examples illustrate specific embodiments and do not limit the scope of the disclosure.

Example 1 : Modeling of the interaction of SARS CoV-2 spike protein and SOR-C13 [0089] The peptide S0R-C13 was modeled to the open and closed (or down) conformations of SARS CoV-2 Spike protein using the MOBYLE bioinformatics portal for analysis of peptide/protein docking. Molecular modeling of the various peptides docking with the Receptor Binding Domain of Spike protein was done with the MOBYLE modeling system located at the RPBS Web Portal hosted at the University of Paris. https://mobyle.rpbs.univ-paris-diderot.fr/cgi-bin/portal.py (Alland et al., 2005; Neron et al., 2009; Saladin et al., 2014).

[0090] Figure 1 illustrates the two conformations available to one of the identical 3 subunits of the Spike protein of SARS CoV-2. The 3-D structure of the Spike trimer at 2.80 A resolution was determined by Walls et al. 2020, using cryo-electron microscopy. The tip of the open conformation of the Spike protein interacts with ACE2 on the surface of cells.

[0091] The output of the MOBYLE docking simulation is shown in Figure 2. The 50 best interactions with the two conformations of the Spike protein are shown. It is clear that folding open the conformation of the subunit reveals a significant interaction domain on the inner face of Spike protein. There are a number of interaction sites on the closed conformation but, as shown below, these show a lower ‘probability’ of binding SOR-C13.

[0092] The MOBYLE peptide docking system does not produce estimates of interaction energies for the binding of peptide to protein. Instead, the system gives what is essentially a measure of the probability of binding in an Interaction Propensity score. This value is the percentage of times any particular amino acid residue of the Spike protein interacts with the 50 best peptide conformations. For example, a value of 60 for amino acid-400 would indicate that amino acid-400 interacted with 60% of the 50 best peptide conformations. As shown in Figure 4, the values of this interaction parameter for the open conformation are about 3-fold larger than for the closed conformation. Only the tip of the one open conformation of Spike, that interacts with ACE2, shows significant interaction with SOR-C13. In other words, this model shows SOR-C13 binds to the same region of Spike protein that interacts with the SARS CoV-2 receptor, ACE2 (See Figure 3).

[0093] The docking of SOR-C13 with open conformation of Spike protein appears to be specific to SARS CoV-2 since SARS CoV-1 open conformation Spike protein showed no significant interaction with SOR-C13 in the Receptor Binding Domain (See Figure 5). [0094] After determining that S0R-C13 appeared to bind specifically to the RBD of the open conformation of the SARS CoV-2 Spike protein (and not the CoV-1 spike) we examined the docking models for a number of other peptides of varying length. The set of interaction graphs following shows the propensity for docking for SOR-C27, SOR-C13, SOR-C11 , SOR-C9, SOR-C7 and SOR-C5; see Table 1 . The region of the Spike protein that interacts with ACE2 is around amino acids 490 to 500 (indicated by the short bar above the propensity plot for the SOR-C13 interaction in Figure 6).

Table 1 : C-terminal soricidin peptide sequences and SEQ ID NOs.

[0095] Figure 7 shows that the amino acids in the site near the RBD of Spike that interacted with C-peptides of different lengths.

[0096] The percentage of times the peptide ligand interacts with a specific region of a protein is a measure of the specificity of the peptide binding to a specific patch on the surface of the protein. Figure 8 shows the percentage of 200 different peptide conformations that interact with the region proximal to the ACE2 binding face. SOR-C13 showed the greatest number of 200 conformations that interacted with the RBD site (-80%) as compared to more ‘non-specific’ interactions elsewhere on the surface of the Spike proteins.

[0097] Assumptions in this model that constrain the outcome are as follows: there are only two interacting structures that interact; the peptide binds to the larger protein complex; the protein does not change conformation when binding the peptide. [0098] The docking simulations outlined here show that a number of C-terminal Soricidin peptides unexpectedly bind specifically to SARS CoV-2 and selectively to the receptor binding domain of the open conformation of the Spike protein.

Example 2: Inhibition of SARS CoV-2 Spike protein interaction with ACE2

[0099] A binding immunoassay was used to examine various C-terminal Soricidin peptides and whether they inhibited the interaction between SARS CoV-2 Spike protein interaction and ACE2. Briefly, SARS CoV-2 spike protein was pre-incubated in tubes with or without the test peptides or a control. An antibody to Spike protein that blocks interaction between Spike protein of SARS CoV-19 and ACE2 (Angiotensin Converting Enzyme 2) was used as a positive control (available from BPS Bioscience, catalogue number 7999). Pre-incubated spike protein +/- test peptide or the control was added to 96-well plates pre-coated with Human ACE2. After washing, bound spike protein was detected using an anti-spike protein antibody reporter (HRP). Reporter signal is inversely proportional to ACE2-spike protein binding interference.

[00100] As shown in Figure 9, the acetate salt of SOR-C13 (SEQ ID NO: 1) and the trifluoroacetate salts of SOR-C9 (SEQ ID NO: 3), SOR-C6 (SEQ ID NO: 4), and SOR-C5 (SEQ ID NO: 5) inhibited the SARS CoV-2 spike/ACE2 interaction. As shown in Figure 10, the acetate salt of SOR-C13 inhibits the SARS CoV-2 spike/ACE2 interaction with an IC50 of 4.5 uM.

Example 3: Inhibition of inflammasome activation and/or activity

[00101] An assay for measuring cellular response to SARS CoV-2 is used to demonstrate the effects of treatment with the peptides described herein on the inflammasome activation and/or activity. Cells are exposed to SARS CoV-2 or one or more SARS CoV-2 proteins in the presence or absence of the peptide. Cellular response is determined by measuring the expression levels or activity of downstream effector molecules such as IL-10, IL-6, IL-7, CSF, IP-10, MCMP-1 , MIP-1-a, TNF-a and/or CXCL10. Expression levels are measured by real time PCR and/or ELISA. T reatment with one or more of the peptides described herein is observed to decrease the expression levels of IL-1 and/or CXCL10 in response SARS CoV-2. References:

Alland C, Moreews F, Boens D, Carpentier M, Chiusa S, Lonquety M, et al. RPBS: a web resource for structural bioinformatics. Nucleic Acids Res. 2005; 33: W44-9.

Bowen CV, DeBay D, Ewart HS, Gallant P, Gormley S, llenchuk TT, et al. In vivo detection of human TRPV6-rich tumors with anti-cancer peptides derived from soricidin. PLoS One. 2013; 8: e58866.

Cavalli, G et al. Interleukin-1 blockade with high-dose anakinra in patients with COVID-19, acute respiratory distress syndrome, and hyperinflammation: a retrospective cohort study. The Lancet. May 7, 2020. DOI:https://doi.org/10.1016/S2665-9913(20)30127-2

Klopfenstein T, Zayet S, Lohse A, Balblanc JC, Badie J, Royer PY, et al. Tocilizumab therapy reduced intensive care unit admissions and/or mortality in COVID-19 patients. Med Mai Infect. 2020.

Neron B, Menager H, Maufrais C, Joly N, Maupetit J, Letort S, et al. Mobyle: a new full web bioinformatics framework. Bioinformatics. 2009; 25: 3005-11.

Nieto-Torres JL, Verdia-Baguena C, Jimenez-Guardeno JM, Regla-Nava JA, Castano-Rodriguez C, Fernandez-Delgado R, et al. Severe acute respiratory syndrome coronavirus E protein transports calcium ions and activates the NLRP3 inflammasome. Virology. 2015; 485: 330-9.

Saladin A, Rey J, Thevenet P, Zacharias M, Moray G, Tuffery P. PEP-SiteFinder: a tool for the blind identification of peptide binding sites on protein surfaces. Nucleic Acids Res. 2014; 42: W221-6.

Walls AC, Park YJ, Tortorici MA, Wall A, McGuire AT, Veesler D. Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein. Cell. 2020; 181 : 281-92 e6.

Claims

CLAIMS:

1. A method of treating or preventing SARS CoV-2 infection in a subject in need thereof, the method comprising administering to the subject an effective amount of a peptide comprising all of part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1 ).

2. The method of claim 1 , wherein the peptide inhibits binding of SARS CoV-2 spike protein to human angiotensin converting enzyme 2 (ACE2).

3. The method of claim 1 or 2, wherein the peptide binds to the SARS CoV-2 spike protein, optionally the ACE2 receptor-binding domain of the SARS CoV-2 spike protein.

4. The method of any one of claims 1 to 3, wherein the peptide suppresses an inflammatory response to SARS CoV-2 infection in the subject.

5. The method of any one of claims 1 to 4, wherein the peptide inhibits Transient Receptor Potential Vanilloid subfamily member s (TRPV6) calcium channel activity.

6. The method of claim 5, wherein the peptide reduces cytoplasmic calcium influx and/or reduces activation or activity of the inflammasome.

7. The method of any one of claims 1 to 6, wherein the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

8. The method of any one of claims 1 to 6, wherein the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

9. The method of any one of claims 1 to 6, wherein the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1 ), optionally wherein the peptide is in the form of an acetate salt.

10. The method of any one of claims 1 to 6, wherein the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

11 . The method of an one of claims 1 to 6, wherein the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

12. A method for inhibiting binding of SARS-CoV-2 spike protein to a cell expressing human angiotensin converting enzyme 2 (ACE2), the method comprising exposing the cell to a peptide comprising all of part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1).

13. The method of claim 12, wherein the peptide binds to the SARS CoV-2 spike protein, optionally the ACE2 receptor-binding domain of the SARS-CoV-2 spike protein.

14. The method of claim 12 or 13, wherein the cell is in vivo, in vitro or ex vivo.

15. The method of any one of claims 12 to 14, wherein the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

16. The method of any one of claims 12 to 14, wherein the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

17. The method of any one of claims 12 to 14, wherein the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1), optionally wherein the peptide is in the form of an acetate salt.

18. The method of any one of claims 12 to 14, wherein the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), or VDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

19. The method of any one of claims 12 to 14, wherein the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

20. The method of any one of claims 1 to 11 , wherein the subject has, or is suspected of having, a SARS CoV-2 infection.

21. The method of claim 20, wherein the subject has been diagnosed with a SARS CoV-2 infection.

22. Use of an effective amount of a peptide comprising all of part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO:1) for treating or preventing SARS CoV-2 infection in a subject in need thereof.

23. Use of an effective amount of a peptide comprising all or part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NOU ) in the manufacture of a medicament for treating or preventing SARS CoV-2 infection in a subject in need thereof.

24. The use of claim 22 or claim 23, wherein the peptide inhibits binding of SARS CoV- 2 spike protein to human angiotensin converting enzyme 2 (ACE2).

25. The use of any one of claims 22 to 24, wherein the peptide binds to the SARS CoV- 2 spike protein, optionally the ACE2 receptor-binding domain of the SARS CoV-2 spike protein.

26. The use of any one of claims 22 to 25, wherein the peptide suppresses an inflammatory response to SARS CoV-2 infection in the subject.

27. The use of any one of claims 22 to 26, wherein the peptide inhibits Transient Receptor Potential Vanilloid subfamily member 6 (TRPV6) calcium channel activity.

28. The use of claim 27, wherein the peptide reduces cytoplasmic calcium influx and/or reduces activation or activity of the inflammasome.

29. The use of any one of claims 22 to 28, wherein the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

30. The use of any one of claims 22 to 28, wherein the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

31 . The use of any one of claims 22 to 28, wherein the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1 ), optionally wherein the peptide is in the form of an acetate salt.

32. The use of any one of claims 22 to 28, wherein the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

33. The use of an one of claims 22 to 28, wherein the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.

34. The use of any one of claims 22 to 33, wherein the subject has, or is suspected of having, a SARS CoV-2 infection.

35. The use of claim 34, wherein the subject has been diagnosed with a SARS CoV-2 infection.

36. Use of a peptide comprising all of part of the amino acid sequence KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1) for inhibiting binding of SARS-CoV-2 spike protein to a cell expressing human angiotensin converting enzyme 2 (ACE2).

37. The use of claim 36, wherein the peptide binds to the SARS CoV-2 spike protein, optionally the ACE2 receptor-binding domain of the SARS-CoV-2 spike protein.

38. The use of claim 36 or 37, wherein the cell is in vivo, in vitro or ex vivo.

39. The use of any one of claims 36 to 38, wherein the peptide comprises between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

40. The use of any one of claims 36 to 38, wherein the peptide consists of between 5 and 13 contiguous amino acids of SEQ ID NO: 1 , optionally between 5 and 13 contiguous amino acids of the C-terminus of SEQ ID NO: 1 .

41 . The use of any one of claims 36 to 38, wherein the peptide comprises or consists of KEFLHPSKVDLPR (SOR-C13; SEQ ID NO: 1 ), optionally wherein the peptide is in the form of an acetate salt.

42. The use of any one of claims 36 to 38, wherein the peptide comprises or consists of HPSKVDLPR (SOR-C9; SEQ ID NO: 3), KVDLPR (SOR-C6; SEQ ID NO: 4), orVDLPR (SOR-C5; SEQ ID NO: 5), optionally wherein the peptide is in the form of a trifluoroacetate (TFA) salt.

43. The use of any one of claims 36 to 38, wherein the peptide comprises an amino acid sequence with at least 70%, 75%, 80% 85%, 90% or 95% identity to SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.