Compounds for use in the treatment of enveloped virus infections - Patents.com

Abstract

The present invention provides a compound for use in treating an enveloped virus infection in a subject, said compound being a compound of formula (I): AA-AA-AA-XYZ(I) as defined herein.The present invention further provides a method of treating an enveloped virus infection in a subject, comprising administering to a subject in need thereof an effective amount of a compound of formula (I).

Description

[Technical field]

[0001] The present invention relates generally to the treatment of certain viral infections. In particular, the present invention relates to the use of certain compounds for the treatment of enveloped virus infections. [Background technology]

[0002] Viruses are infectious agents that can only replicate within a host organism. Viruses can infect a variety of living organisms, including humans. When virus particles are independent of their host cells, they typically contain the viral genome (which may be DNA or RNA, single- or double-stranded, linear or circular) contained within a protein shell called a capsid. In some viruses, called enveloped viruses, the protein shell is enclosed within a membrane called the envelope. Other viruses are non-enveloped.

[0003] Viral infections represent a significant healthcare problem. Over the past two decades, the world has seen a rise in the incidence of viral infections. For example, in 2020, the SARS-CoV-2 virus (Severe Acute Respiratory Syndrome Coronavirus 2), an enveloped virus that causes COVID-19, forced many parts of the world into "lockdown" with severe and long-lasting consequences for both the global economy and international health. It is of great importance to identify new therapeutic and protective treatments against such viruses.

[0004] There are many different types of antiviral agents, such as entry inhibitors, uncoating inhibitors, release (or exit) inhibitors, protease inhibitors, and nucleotide / nucleoside analogues. As an example, the small molecule oseltamivir, sold under the trade name Tamiflu, is a neuraminidase inhibitor that inhibits the release of influenza A and B viruses (which are enveloped viruses) from host cells. Viral resistance to antiviral agents is a significant problem in global health care. In this regard, for example, mutations in viral proteins can make such mutated viruses resistant to treatment with antiviral drugs that act by targeting the viral proteins. For example, mutant influenza A viruses that are resistant to oseltamivir have been reported. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] WO2009 / 081152A2 [Non-patent literature]

[0006] [Non-Patent Document 1] Saravolatz et al., Antimicrobial Agents and Chemotherapy (2012), vol. 56(8), pp. 4478-4482 [Non-Patent Document 2] Chapter 14 of Drug Design and Development, Krogsgaard, Larsen, Liljefors and Madsen (eds.) 1996, Horwood Acad. Pub. [Non-Patent Document 3] Schmidt, R. et al., Int. J. Peptide Protein Res., 1995, 46, 47 [Non-Patent Document 4] Chorev, M. and Goodman, M., Acc. Chem. Res, 1993, 26, 266 [Non-Patent Document 5] Sherman DB and Spatola, AFJ Am. Chem. Soc., 1990, 112, 433. [Non-Patent Document 6] Hoffman, RV and Kim, HOJ Org. Chem., 1995, 60, 5107 [Non-Patent Document 7] Allmendinger, T. et al., Tetrahydron Lett., 1990, 31, 7297 [Non-Patent Document 8] Sasaki, Y. and Abe, J. Chem. Pharm. Bull. 1997 45, 13 [Non-Patent Document 9] Spatola, AF, Methods Neurosci, 1993, 13, 19 [Non-Patent Document 10] Lavielle, S. et al., Int. J. Peptide Protein Res., 1993, 42, 270 [Non-Patent Document 11] Luisi, G. et al., Tetrahedron Lett. 1993, 34, 2391 [Non-Patent Document 12] Ostresh, JM et al., Proc. Natl. Acad. Sci. USA (1994) 91, 11138~11142 [Non-Patent Document 13] L. J. Reed and H. Muench, American Journal of Epidemiology, Vol. 27, No. 3, 1938, pp. 493-497 Summary of the Invention [Problem to be solved by the invention]

[0007] It is clear that alternative, and preferably advantageous, anti-viral treatments (especially against viruses that cause disease in humans) would be highly desirable. Such treatments would be useful in treating or preventing infections (e.g., in humans) with viral pathogens. [Means for solving the problem]

[0008] The present inventors have surprisingly found that a class of tripeptide compounds having certain C-terminal modifications exhibit superior antiviral activity against enveloped viruses, including against enveloped viruses pathogenic to humans. Such tripeptides are cationic (positively charged) and bulky. One compound in this class is the compound LTX-109. LTX-109 has previously been reported to exhibit antimicrobial activity (e.g., Saravolatz et al., Antimicrobial Agents and Chemotherapy (2012), Vol. 56(8) pp. 4478-4482), but antiviral activity of these molecules has not been previously demonstrated. In light of the present inventors' findings, it is clear that such compounds are an important class of agents to add to the current arsenal of anti-enveloped virus therapies.

[0009] Thus, in one aspect, the present invention provides a compound for use in treating an enveloped virus infection in a subject, the compound being of formula (I) AA-AA-AA-XYZ (I) wherein, in any order, two of the AA (amino acid) moieties are cationic amino acids, preferably lysine or arginine, but may also be histidine or any non-genetically encoded or modified amino acid having a positive charge at pH 7.0, one of the AA is an amino acid with a large lipophilic R group, the R group having 14-27 non-hydrogen atoms, preferably containing two or more, e.g. 2 or 3, cyclic groups which may be fused or connected, these cyclic groups will typically contain 5 or 6 non-hydrogen atoms, preferably 6 non-hydrogen atoms (in the case of fused rings non-hydrogen atoms may of course be shared), X is a branched or unbranched C1-C 10 an N atom which may, but preferably is not, substituted by an alkyl or aryl group, such as methyl, ethyl or phenyl, which group may incorporate up to two heteroatoms selected from N, O and S, Y is -R a -R b -, -R a -R b -R b -and-R b -R b -R a -, wherein R a is C, O, S or N, preferably C; R b is C and R a and R b Each of may be substituted or unsubstituted with a C1-C4 alkyl group, and preferably Y is -R a -R b - (where R a is preferably C, preferably this group is unsubstituted and Y is -R a -R b -R b -or-R b -R b -R a -, preferably R a and R b is substituted with one or more of Z is a group comprising 1 to 3 cyclic groups of 5 or 6 non-hydrogen atoms each (preferably C atoms), two or more of the cyclic groups may be fused and one or more of the rings may be substituted, which substitutions may but typically do not include polar groups, suitable substituents include halogen, preferably bromine or fluorine, and C1-C4 alkyl groups, the Z moiety incorporates up to 15, preferably 5 to 12 non-hydrogen atoms, most preferably it is phenyl; The bond between Y and Z is the R of Y. a or R b and a non-hydrogen atom of one of the cyclic groups of Z. The present invention provides a compound,

[0010] Suitable non-genetically encoded and modified amino acids which can provide cationic amino acids include analogs of lysine, arginine and histidine, such as homolysine, ornithine, diaminobutyric acid, diaminopimelic acid, diaminopropionic acid and homoarginine, as well as trimethylysine and trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-carbamimidoylpiperidine-4-carboxylic acid and 4-guanidinophenylalanine.

[0011] The large lipophilic R group of AA may contain heteroatoms such as O, N or S, typically there will be no more than one heteroatom, preferably nitrogen. This R group will preferably have no more than two polar groups, more preferably none or one, most preferably none.

[0012] The compounds for use according to the present invention are preferably peptides.

[0013] The compound for use according to the invention preferably has the formula (II) AA1-AA2-AA1-XYZ (II) [In the formula, AA1 is a cationic amino acid, preferably lysine or arginine, but may also be histidine or any non-genetically encoded or modified amino acid that has a positive charge at pH 7.0; AA2 is an amino acid with a large lipophilic R group, the R group having 14-27 non-hydrogen atoms, preferably containing two or more, e.g. two or three, cyclic groups which may be fused or connected, and these cyclic groups will typically contain 5 or 6 non-hydrogen atoms, preferably 6 non-hydrogen atoms; X, Y and Z are as defined above. It is of the following.

[0014] Further preferred compounds for use according to the present invention are compounds of formula (III) and (IV): AA2-AA1-AA1-XYZ (III) AA1-AA1-AA2-XYZ (IV) [In the formula, AA1, AA2, X, Y and Z are as defined above.] More preferred are molecules of formula (II):

[0015] Among the above compounds, certain ones are particularly preferred. In particular, the amino acid with a large lipophilic R group, conveniently referred to herein as AA2, is tributyltryptophan (Tbt) or a biphenylalanine derivative, such as Phe(4-(2-naphthyl)), Phe(4-(1-naphthyl)), Bip(4-n-Bu), Bip(4-Ph) or Bip(4-T-Bu), with Phe(4-(2-naphthyl)) and Tbt being most preferred. In some preferred embodiments, the amino acid with a lipophilic R group is tributyltryptophan (Tbt).

[0016] In some preferred embodiments, Y is -R a -R b - and unsubstituted, most preferably R a and R b are all carbon (C) atoms. Preferably, Y is -CH2-CH2-.

[0017] In some preferred embodiments, Z is phenyl (Ph).

[0018] A further preferred group of compounds is where -XYZ together are a -NHCH2CH2Ph ​​group.

[0019] The compounds include all enantiomeric forms, both D and L amino acids, as well as enantiomers arising from chiral centers in the amino acid R group and the C-terminal capping group "-XYZ". Included within the term "amino acid" are beta and gamma amino acids as well as alpha amino acids, all of which are N-substituted glycines that may be considered AA units. Compounds for use in accordance with the invention include beta peptides and depsipeptides.

[0020] The most preferred compounds have the structural formula: [ka] where t-Bu represents a tertiary butyl group. This compound having the above structural formula incorporating the amino acid 2,5,7-tris-tert-butyl-L-tryptophan is the most preferred compound for use in the present invention (also referred to herein as LTX-109). Analogs of this compound incorporating other cationic residues, particularly Lys, in place of Arg are also highly preferred. Analogs incorporating alternative C-terminal capping groups as defined above are also highly preferred.

[0021] Further preferred compounds for use according to the present invention are [ka] This compound (i.e., the compound having the structural formula depicted immediately above) may be referred to as Arg-Phe(4-(1-naphthyl))-Arg-NH-CH2-CH2-Ph. This compound is a compound of formula (II) where AA1 is arginine (Arg), AA2 is Phe(4-(1-naphthyl)), and -XYZ together are the group -NHCH2CH2Ph.

[0022] Further preferred compounds for use according to the present invention are [ka] This compound (i.e., the compound having the structural formula depicted immediately above) may be referred to as Arg-Phe(4-(2-naphthyl))-Arg-NH-CH2-CH2-Ph. This compound is also referred to herein as LTX-7. This compound is of formula (II) where AA1 is arginine (Arg), AA2 is Phe(4-(2-naphthyl)), and -XYZ together are a -NHCH2CH2Ph ​​group.

[0023] Further preferred compounds for use according to the present invention are [ka] where t-Bu represents a tertiary butyl group. This compound (i.e., a compound having the structural formula depicted immediately above) may be referred to as Lys-Tbt-Lys-NH-CH2-CH2-Ph. This compound is also referred to herein as LTX-12. This compound is a compound of formula (II) where AA1 is lysine (Lys), AA2 is tributyltryptophan (Tbt, which may also be referred to as 2,5,7-tris-tert-butyl-L-tryptophan), and -XYZ together are a -NHCH2CH2Ph ​​group.

[0024] In a preferred embodiment, the compound for use according to the present invention is selected from the group consisting of LTX-109, LTX-7 and LTX-12. The compound LTX-109 is the most preferred compound for use according to the present invention.

[0025] The compounds for use in the present invention are preferably peptides.

[0026] The compounds of formulae (I) through (IV) may be peptidomimetics, and peptidomimetics of the peptides described and defined herein also represent compounds for use in accordance with the present invention. Peptidomimetics are typically characterized by retaining the polarity, three-dimensional size, and functionality (biological activity) of their peptide equivalents, but where the peptide bond is replaced by a linkage that is often more stable. By "stable" we mean more resistant to enzymatic degradation by hydrolases. In general, bonds that replace amide bonds (amide bond surrogates) preserve many of the properties of the amide bond, such as conformation, steric bulk, electrostatic characteristics, hydrogen bonding potential, etc. Chapter 14 of "Drug Design and Development," Krogsgaard, Larsen, Liljefors, and Madsen (eds.) 1996, Horwood Acad. Pub, provides a general review of techniques for the design and synthesis of peptidomimetics. In the present case, where a molecule can react with a membrane rather than with a specific active site of an enzyme, some of the described problems of closely mimicking affinity and efficacy or substrate function are irrelevant and peptidomimetics can be easily prepared based on a given peptide structure or motif of the required functional groups.Suitable amide bond surrogates include the following groups: N-alkylated (Schmidt, R. et al., Int. J. Peptide Protein Res., 1995, 46, 47), retro-inverse amide (Chorev, M and Goodman, M., Acc. Chem. Res., 1993, 26, 266), thioamide (Sherman DB and Spatola, AFJ Am. Chem. Soc., 1990, 112, 433), thioester, phosphonate, ketomethylene (Hoffman, RV and Kim, HOJ Org. Chem., 1995, 60, 5107), hydroxymethylene, fluorovinyl (Allmendinger, T. et al., Tetrahydron Lett., 1990, 31, 7297), vinyl, methyleneamino (Sasaki, Y and Abe, J. Chem. Pharm. Bull. 1997 45, 13), methylenethio (Spatola, AF, Methods Neurosci, 1993, 13, 19), alkanes (Lavielle, S. et al., Int. J. Peptide Protein Res., 1993, 42, 270) and sulfonamides (Luisi, G. et al., Tetrahedron Lett. 1993, 34, 2391).

[0027] The peptidomimetic compounds used in the present invention will typically have three distinct subunits that are roughly equivalent in size and function to amino acids (AA units).Therefore, the term "amino acid" may be used herein for convenience to refer to the equivalent subunits of peptidomimetic compounds.In addition, peptidomimetics may have groups equivalent to the R groups of amino acids, and the discussion herein of suitable R groups and N- and C-terminal modification groups applies mutatis mutandis to peptidomimetic compounds.

[0028] As discussed in the above referenced textbooks, as well as replacing amide bonds, peptidomimetics may replace larger structural moieties with dipeptide or tripeptidomimetic structures, in which case mimetic moieties with peptide bonds, such as azole-derived mimetics, may be used as dipeptide replacements. However, peptidomimetics in which the amide bonds are replaced as discussed above and thus peptidomimetic backbones are preferred.

[0029] Suitable peptidomimetics include reduced peptides in which the amide bonds have been reduced to methylene amines by treatment with reducing agents such as borane or hydride reagents such as lithium aluminum hydride. Such reduction has the added benefit of increasing the overall cationic character of the molecule.

[0030] Other peptidomimetics include, for example, peptoids formed by stepwise synthesis of amide-functionalized polyglycines. Some peptidomimetic scaffolds are readily accessible from their peptide precursors, for example permethylated peptides, and a suitable method is described by Ostresh, JM et al., Proc. Natl. Acad. Sci. USA (1994) 91, 11138-11142. Strongly basic conditions favor N-methylation over O-methylation, resulting in methylation of some or all of the nitrogen atoms at the peptide bond and the N-terminal nitrogen.

[0031] Preferred peptidomimetic backbones include polyesters, polyamines and their derivatives, as well as substituted alkanes and alkenes. Peptidomimetics will preferably have N- and C-termini which may be modified as discussed herein.

[0032] The compounds for use in the present invention can be synthesized in any convenient manner. Typically, reactive groups present (e.g., amino, thiol and / or carboxyl) will be protected throughout the entire synthesis. Thus, the final step in the synthesis will be the deprotection of the protected derivatives of the present invention.

[0033] In constructing the peptides it is in principle possible to start either from the C-terminus or from the N-terminus, although the C-terminal starting procedure is preferred.

[0034] Methods of peptide synthesis are well known in the art, however, in the present invention it may be particularly convenient to carry out the synthesis on a solid phase support, such supports being well known in the art.

[0035] A wide choice of protecting groups for amino acids is known, and suitable amine protecting groups may include carbobenzoxy (also designated Z), t-butoxycarbonyl (also designated Boc), 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr) and 9-fluorenylmethoxy-carbonyl (also designated Fmoc). It will be appreciated that when peptides are constructed from the C-terminus, an amine protecting group is present on the α-amino group of each new residue added and must be selectively removed prior to the next coupling step.

[0036] For example, carboxyl protecting groups that can be used include easily cleaved ester groups, such as benzyl (Bzl), p-nitrobenzyl (ONb), pentachlorophenyl (OPCIP), pentafluorophenyl (OPfp) or t-butyl (OtBu) groups, and methyl groups which are linked to a coupling group on a solid support, such as polystyrene.

[0037] Thiol protecting groups include p-methoxybenzyl (Mob), trityl (Trt) and acetamidomethyl (Acm).

[0038] A wide range of procedures exist for the removal of amine and carboxyl protecting groups; however, these must be consistent with the synthetic strategy employed. The side chain protecting groups must be stable to the conditions used to remove the temporary α-amino protecting group prior to the next coupling step.

[0039] Amine protecting groups, such as Boc, and carboxyl protecting groups, such as tBu, can be simultaneously removed by acid treatment, for example with trifluoroacetic acid. Thiol protecting groups, such as Trt, can be selectively removed using an oxidizing agent, for example iodine.

[0040] Compounds for use according to the present invention (eg, LTX-109) may be synthesized as described in WO2009 / 081152A2.

[0041] The compounds (e.g., peptides) for use according to the invention exhibit activity against enveloped viruses, in other words, the compounds for use according to the invention exhibit anti-enveloped virus activity.

[0042] The compounds used in the present invention typically exhibit activity against enveloped viruses (anti-enveloped virus activity) in (or determined or assessed by) a suitable in vitro assay, such as an end-point dilution assay (e.g., TCID50 assay). The skilled artisan is familiar with suitable in vitro assays, such as a suitable end-point dilution assay (e.g., TCID50 assay). A preferred TCID50 assay is described in the Examples section herein. The compounds used in the present invention may exhibit activity against enveloped viruses (anti-enveloped virus activity) determined (or assessed by) microscopy, such as electron microscopy. The compounds used in the present invention may cause viral envelope disruption (or envelope destabilization or lysis) by any suitable means or assay, such as assessed (or determined by) microscopy, such as electron microscopy. A preferred electron microscopy method is described in Example 1 herein.

[0043] The compounds for use in the present invention can exert their anti-envelope virus effects through mechanisms that directly affect the membrane (or viral envelope) and therefore can be considered as membrane (or viral envelope) acting antivirals. These compounds can therefore be considered to cause lysis, destabilize or even penetrate the viral envelope. This confers a clear therapeutic advantage over agents that act on or interact with proteinaceous components of the target virus. Mutations in viral proteins can result in new forms of viral proteins that lead to resistance to antiviral agents that act by targeting such viral proteins. However, the development of resistance is less of an issue when the target is a lipid layer (or lipid membrane) (derived from the host cell) as opposed to a specific viral protein target. The envelope disrupting effect can cause very rapid destruction of enveloped virus particles. In addition to direct membrane (envelope) disrupting or destabilizing activity, the compounds for use in accordance with the present invention can have other useful properties that destroy or inhibit the target virus (e.g., by other mechanisms of action).

[0044] As indicated above, the present invention provides a compound as defined elsewhere herein for use in treating an enveloped virus infection. In other words, the present invention provides a compound as defined herein for use in treating an infection in a subject, wherein the causative agent of said infection is an enveloped virus.

[0045] An "enveloped virus" is a virus that is enveloped (or wrapped) in a lipid layer (or lipid membrane). The lipid layer may be a lipid bilayer. Thus, an enveloped virus has a capsid (viral capsid) that is covered (or enveloped or wrapped or surrounded by) an outer membrane or envelope that comprises a lipid layer (typically a phospholipid layer), such as a lipid bilayer. The viral envelope may also comprise one or more virally encoded proteins (e.g., glycoproteins). The lipid layer of the viral envelope is derived (or obtained) from the lipid membrane (e.g., lipid bilayer) of the infected host cell. Of course, the enveloped virus according to the present invention has an envelope from (or derived or obtained) from a eukaryotic lipid membrane, preferably from a mammalian (e.g., human) lipid membrane. Such eukaryotic lipid membranes may be the cell membrane or the membrane of a cell organelle (e.g., the endoplasmic reticulum; or the Golgi apparatus (or Golgi apparatus); or the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), sometimes called vesicle-tubule cluster (VTC)). The viral envelope is typically acquired at the membrane (lipid bilayer) of the host cell in a process that may be called "budding" or "budding shedding". During the budding process, the newly formed virus particle is "wrapped" (or "surrounded" or "coated") within an envelope made of the lipid membrane of the host cell (the virus's host cell). Thus, the lipid layer (or lipid membrane) of the viral envelope may be considered to be directly derived from the host cell (directly derived from the host cell's membrane).

[0046] In some embodiments, an enveloped virus is a virus that has an envelope (viral envelope) that is derived from (or acquired from or characteristic of) a cellular membrane (i.e., the cellular membrane of the virus' host cell). The cellular membrane may also be referred to as the plasma membrane, cytoplasmic membrane, or plasma lemma.

[0047] In some embodiments, an enveloped virus is a virus that has an envelope (viral envelope) that is derived from (or acquired from or characteristic of) a membrane of an intracellular organelle (membrane-bound organelle) (i.e., an organelle membrane of the virus' host cell). Such membrane-bound organelles include, for example, the endoplasmic reticulum, the Golgi apparatus (or Golgi apparatus), or the endoplasmic reticulum-Golgi intermediate compartment (ERGIC).

[0048] Any enveloped virus infection can be treated according to the present invention. Typically and preferably, the enveloped virus is a virus that infects (or can infect) a mammal. Mammals include, for example, humans and any livestock, domestic or laboratory animals. Specific examples include mice, rats, pigs, cats, dogs, sheep, rabbits, cows and monkeys. In some embodiments of the present invention, the mammal is a human. Thus, typically and preferably, the enveloped virus according to the present invention is a mammalian pathogen, preferably a human pathogen.

[0049] In some embodiments, the enveloped virus is the causative agent of a respiratory tract infection. The respiratory tract infection may be an infection of the upper and / or lower respiratory tract. In some embodiments, the respiratory tract infection is an infection of the upper respiratory tract.

[0050] The enveloped virus may be a DNA virus or an RNA virus, hi some embodiments, the enveloped virus is an RNA virus (e.g., a single-stranded (ss) RNA enveloped virus).

[0051] In some embodiments, the enveloped virus may be a virus from one of the following classes: herpesvirus, poxvirus, hepadnavirus, asfarvirus, flavivirus, alphavirus, togavirus, coronavirus, orthomyxovirus, orthopneumovirus, paramyxovirus, rhabdovirus, bunyavirus, filovirus, or retrovirus.

[0052] In some embodiments, the enveloped virus is an orthopneumovirus (e.g., respiratory syncytial virus, RSV), an orthomyxovirus (e.g., an influenza virus, such as influenza A virus), or a coronavirus (e.g., severe acute respiratory syndrome coronavirus 2, SARS-CoV-2).

[0053] In some embodiments, the enveloped virus is an orthopneumovirus (eg, respiratory syncytial virus, RSV).

[0054] In some embodiments, the enveloped virus is an orthomyxovirus (e.g., influenza A virus).

[0055] In some embodiments, the enveloped virus is a coronavirus (e.g., severe acute respiratory syndrome coronavirus 2, SARS-CoV-2).

[0056] In some embodiments, the enveloped virus is selected from the group consisting of respiratory syncytial virus (RSV), influenza A virus, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0057] In some embodiments, the enveloped virus is respiratory syncytial virus (RSV). RSV can cause infections of the respiratory tract, for example in humans. In some embodiments, the enveloped virus is not RSV.

[0058] In some embodiments, the enveloped virus is influenza A virus. Influenza A can cause infection of the respiratory tract, for example, in humans. In some embodiments, the enveloped virus is not influenza A virus.

[0059] In some embodiments, the enveloped virus is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 can cause infection of the respiratory tract, for example, in humans. SARS-CoV-2 is a virus that can cause coronavirus disease 2019 (COVID-19). In some embodiments, the enveloped virus is not a SARS-CoV-2 virus.

[0060] In an alternative view, in one aspect the invention provides a compound as defined herein for use in treating a disease or condition caused by an enveloped virus infection. The embodiments of the other aspects of the invention described herein apply mutatis mutandis to this aspect of the invention.

[0061] In some embodiments, the disease or condition to be treated is a respiratory tract infection (eg, an upper and / or lower respiratory tract infection).

[0062] In some embodiments, the disease or condition being treated is influenza.

[0063] In some embodiments, the disease or condition being treated is coronavirus disease 2019 (COVID-19).

[0064] The compounds for use according to the present invention are typically presented (or administered) in the form of a formulation or composition comprising one or more compounds according to the present invention in admixture with suitable diluents, carriers and / or excipients. Suitable diluents, excipients and carriers are known to those skilled in the art. Thus, the present invention provides a formulation (or composition) comprising a compound as defined herein for use in treating enveloped virus infections. Of course, typically and preferably, the formulation (or composition) is a pharmaceutical formulation (or pharmaceutical composition). Thus, preferably, the diluent, carrier and / or excipient is a pharmaceutically acceptable diluent carrier and / or carrier.

[0065] Compositions for use according to the invention may be presented in a form suitable for, for example, oral, nasal, respiratory tract (e.g., upper respiratory tract), parenteral, intravenous, topical or rectal administration. A person skilled in the art can readily select an appropriate administration form, for example based on the type (or location) of infection to be treated.

[0066] The compounds (or formulations or compositions) for use according to the present invention may be administered orally, nasally, parenterally, intravenously, topically or rectally.

[0067] A compound (or formulation or composition) for use according to the invention may be administered to the respiratory tract, for example the upper respiratory tract.

[0068] As used herein, the term "pharmaceutical" includes veterinary applications of the present invention.

[0069] The active compounds defined herein may be presented in conventional pharmaceutical dosage forms, such as tablets, coated tablets, solutions, emulsions, liposomes, powders, capsules or sustained release forms.

[0070] Conventional pharmaceutical excipients and normal manufacturing methods may be used for preparing these forms.

[0071] Tablets can, for example, be produced by mixing the active ingredient(s) with known excipients, such as diluents, for example calcium carbonate, calcium phosphate or lactose, disintegrating agents, for example corn starch or alginic acid, binders, for example starch or gelatin, lubricants, for example magnesium stearate or talcum, and / or agents for obtaining sustained release, for example carboxypolymethylene, carboxymethylcellulose, cellulose acetate phthalate or polyvinyl acetate.

[0072] The tablet may, if desired, consist of several layers. Coated tablets may be produced by coating a core obtained in a manner similar to that of the tablet with an agent commonly used for tablet coating, such as polyvinylpyrrolidone or shellac, gum arabic, talcum, titanium dioxide or sugar. To obtain sustained release or to avoid incompatibilities, the core may also consist of several layers. The tablet coat may consist of several layers to obtain sustained release, in which case the excipients mentioned above for the tablet may be used.

[0073] Solutions (e.g., injection solutions) may, for example, be produced in a conventional manner, for example by the addition of a preservative, for example a p-hydroxybenzoate, or a stabilizer, for example EDTA. The solutions may be filled into vials or ampoules.

[0074] Capsules containing one or several active ingredients can be produced, for example, by mixing the active ingredients with an inert carrier, such as lactose or sorbitol, and filling the mixture into gelatin capsules.

[0075] Suitable suppositories can be produced, for example, by mixing the active ingredient or active ingredient combination with conventional carriers envisaged for this purpose, such as natural fats or polyethylene glycol or their derivatives.

[0076] The dosage may vary based on parameters such as age, weight and sex of the subject. Suitable dosages may be readily established by those skilled in the art. Suitable dosage units may be readily prepared.

[0077] Treatment according to the invention may involve co-administration with one or more additional active agents used in the treatment or prevention of enveloped virus infections (or conditions caused thereby). Generally speaking, one or more additional active agents may be administered to a subject substantially simultaneously with a compound according to the invention, for example from a single pharmaceutical composition or from two pharmaceutical compositions administered closely together. Thus, in some embodiments, a pharmaceutical composition may additionally comprise one or more additional active ingredients (e.g., one or more additional antiviral compounds). Alternatively, one or more additional active agents may be administered to a subject at a time subsequent to administration of a compound according to the invention. "Subsequently at a time" as used herein means "staggered", such that one or more additional agents are administered to a subject at a time different from administration of a compound according to the invention. In general, the two agents will be administered at times effectively spaced apart to allow the two agents to exert their respective therapeutic effects, i.e., they are administered at a "biologically effective time interval". The one or more additional active agents may be administered to the subject at a biologically effective time prior to the compounds according to the invention or at a biologically effective time following the compounds according to the invention.

[0078] As used herein, the term "treatment" or "therapy" includes therapeutic and prophylactic (or prophylactic) therapy. Thus, the compounds for use according to the invention may be for therapeutic or prophylactic use.

[0079] Viewed alternatively, the present invention provides a method of treating an enveloped virus infection in a subject (or patient), comprising administering to a subject in need thereof a therapeutically or protectively effective amount of a compound as defined herein. The embodiments of the invention described herein in relation to the other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0080] The present invention also provides a method of treating a disease or condition caused by (or characterised by) an enveloped virus infection, comprising administering to a patient in need thereof a therapeutically or protectively effective amount of a compound as defined herein. The embodiments of the invention described herein in relation to the other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0081] An effective amount (e.g., a therapeutically or protectively effective amount) will be determined based on clinical evaluation and can be easily monitored. The amount administered should typically be effective to kill or inactivate all or part of the targeted enveloped viruses, or to prevent or reduce their reproduction rate, or otherwise reduce their deleterious effects on the body. Administration may be protective.

[0082] Viewed further still, the present invention provides the use of a compound as defined herein in the manufacture of a medicament for use in the treatment of an enveloped virus infection. The embodiments of the invention described herein in relation to the other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0083] Viewed yet another way, the invention provides the use of a compound as defined herein in the manufacture of a medicament for use in the treatment of a disease or condition caused by (or characterised by) an enveloped virus infection. The embodiments of the invention described herein in relation to other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0084] Viewed yet another way, the present invention provides the use of a compound as defined herein for the treatment of an enveloped virus infection. The embodiments of the invention described herein in relation to the other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0085] Viewed yet another way, the present invention provides the use of a compound as defined herein for the treatment of a disease or condition caused by (or characterised by) an enveloped virus infection. The embodiments of the invention described herein in relation to the other aspects of the invention apply mutatis mutandis to this aspect of the invention.

[0086] In a further aspect there is provided a compound of the invention for use in destabilising and / or permeabilising the envelope of an enveloped virus.

[0087] The term "subject" or "patient" as used herein includes any mammal, such as humans, and any livestock, domestic or laboratory animal. Specific examples include mice, rats, pigs, cats, dogs, sheep, rabbits, cows and monkeys. However, preferably, the subject or patient is a human subject. Thus, the subject or patient to be treated according to the present invention will preferably be a human.

[0088] In some embodiments, the subject according to the present invention is a subject with an enveloped virus infection. In some embodiments, the subject according to the present invention is a subject suspected of having an enveloped virus infection. In some embodiments, the subject according to the present invention may be a subject at risk of developing (or at risk of contracting) an enveloped virus infection.

[0089] In some embodiments, the subject according to the present invention is a subject with a disease or condition caused by an enveloped virus infection. In some embodiments, the subject according to the present invention is a subject suspected of having a disease or condition caused by an enveloped virus infection. In some embodiments, the subject according to the present invention may be a subject at risk of developing (or at risk of suffering from) a disease or condition caused by an enveloped virus infection.

[0090] The invention also provides kits comprising one or more of the compounds according to the invention for use in the methods and uses described herein. Preferably, the kits include instructions for use in treating an enveloped virus infection as described herein.

[0091] As used throughout this application, the terms "a" and "an" are used to mean "at least one," "at least the first," "one or more," or "multiple" of the referenced component or step, unless an upper limit is specifically recited thereafter.

[0092] In addition, when the terms "comprise," "comprises," "has," or "having" or other equivalent terms are used herein, in some more specific embodiments these terms include the terms "consisting of" or "consisting essentially of" or other equivalent terms.

[0093] The invention will now be further described with reference to the following non-limiting examples and figures. [Brief description of the drawings]

[0094] [Figure 1] FIG. 1 shows electron microscopy images of lentivirus-like particles after 10 min incubation with buffer only (control) or 1% LTX-109 solution (1% Peptid). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS EXAMPLES

[0095] Example 1 In vitro effects of 1% LTX-109 on lentivirus-like particle envelope The virus used in this example is a lentivirus-like particle expressing the vesicular stomatitis glycoprotein (VSV-G) on its surface. This virus-like particle behaves like a complete enveloped virus, but does not have a genome. Virus samples were loaded onto glow discharge EM grids by incubating the grids with either control virus (in buffer) or virus plus 1% LTX-109 final solution for 10 minutes. The grids were then dried with filter paper and negative stain (4% Uranyl acetat in water) was added for 2 minutes. After removal of the stain, the grids were briefly dried and then imaged on a JEOL JEM-1230 electron microscope at 80 kV. Images were recorded with a Morada camera and further processed in Adobe Photoshop. The acquired images are shown in Figure 1.

[0096] Figure 1 clearly shows that lentivirus-like particles after control (buffer only) treatment (incubation) have intact envelopes, whereas lentivirus-like particles after 1% LTX-109 treatment (incubation) have disrupted (or lysed) envelopes. Thus, this experiment demonstrates the antiviral membrane disruption / destabilization effect of LTX-109 on lentivirus-like particles.

[0097] Example 2 Antiviral activity of LTX-109 against influenza A virus (IAV) the aim The aim of this study was to test the antiviral activity of LTX-109 against IAV (influenza A virus).

[0098] method To test whether 1% LTX-109 (w / v) has antiviral activity against IAV, 1 × 10 6 Infectious units of IAV (A / WSN / 33; 40 μl) were incubated with 4 volumes of 1% LTX-109 in PBS (160 μl) or PBS (phosphate buffered saline) control. Experiments were performed in triplicate.

[0099] After 1 hour, the incubation was stopped by adding excess cold medium and the formulation was physically separated from the virus through a filter to reduce cytotoxicity in the assay cells. Some turbidity during the preparation was observed for the 1% formulation, but no precipitation. Infectious virus was quantified via serial dilution (a series of 10-fold dilutions) on monolayers of MDCK-II cells in microtiter plates (MDCK-II cells are mammalian cells that can display cytopathic effects (CPE) upon viral infection). The starting solution for the serial dilutions (i.e., the original (or undiluted) solution) was obtained by resuspending the virus, separated via the filtration step, in 1 ml of medium. For each dilution of virus in the dilution series, eight wells of the microtiter plate were tested (i.e., each dilution of virus was applied to eight separate wells, each well containing a monolayer of MDCK-II cells). Appropriate controls were also performed. Five days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the wells at a given dilution) displayed virus-induced cytopathic effects (TCID50). TCID50 (TCID50 / ml) assay (tissue culture infectious dose 50 assay) is a type of end-point dilution assay that is well known in the art and routinely used to quantitatively measure virus titer. TCID50 / ml provides a measure of infectious units of virus / ml. " / ml" refers to / ml of the starting solution (i.e., the neat / undiluted solution) mentioned above.

[0100] A parallel study was included in which the same procedure was performed in the absence of virus to determine any residual cytotoxic effects of the formulation on the assay cells.

[0101] result The results of testing the antiviral activity of 1% LTX-109 against influenza A virus (IAV) are summarized in Table 1 (below).

[0102] After 1 hour of incubation with the PBS control, a mean of 5.16E+04 TCID50 / ml of IAV was measured.

[0103] After 1 hour incubation with 1% LTX-109, an average of 1.58E+01 TCID50 / ml was measured, corresponding to a greater than 3 log or 99.9% reduction in infectivity compared to the PBS control.

[0104] After filtration, cytotoxicity was observed up to the first dilution of the 1% formulation without affecting the validity of the study.

[0105] [Table 1]

[0106] conclusion Based on the findings reported here, exposure of IAV to 1% LTX-109 for 1 h in vitro caused a greater than 3 log decrease in viral infectivity compared to the PBS control, which corresponds to a 99.9% reduction. These results indicate that LTX-109 has excellent antiviral activity against influenza A viruses (enveloped viruses).

[0107] Example 3 Antiviral activity of LTX-109 against respiratory syncytial virus the aim The aim of this study was to test the antiviral activity of LTX-109 against RSV (respiratory syncytial virus).

[0108] method To test whether 1% LTX-109 (w / v) and 0.1% LTX-109 (w / v) have antiviral activity against RSV, 1 × 10 5 Infectious units of RSV (40 μl) were incubated with 4 volumes of 1% or 0.1% LTX-109 in PBS (160 μl) or PBS (phosphate buffered saline) control. Experiments were performed in triplicate.

[0109] After 1 hour, the incubation was stopped by adding excess cold medium and the formulation was physically separated from the virus through a filter to reduce cytotoxicity in the assay cells. Some turbidity during the preparation was observed, especially for the 1% formulation, but no precipitation. Infectious virus was quantified via serial dilution (a series of 10-fold dilutions) on monolayers of Hep2 cells in microtiter plates (Hep2 cells are mammalian cells that can display cytopathic effects (CPE) upon viral infection). The starting solution for the serial dilutions (i.e., the original (or undiluted) solution) was obtained by resuspending the virus, separated via the filtration step, in 1 ml of medium. For each dilution of virus in the dilution series, eight wells of the microtiter plate were tested (i.e., each dilution of virus was applied to eight separate wells, each well containing a Hep2 cell monolayer). Appropriate controls were also performed. Eight days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the wells at a given dilution) displayed virus-induced cytopathic effects (TCID50). TCID50 (TCID50 / ml) assay (tissue culture infectious dose 50 assay) is a type of end-point dilution assay that is well known in the art and routinely used to quantitatively measure virus titer. TCID50 / ml provides a measure of infectious units of virus / ml. " / ml" refers to / ml of the starting solution (i.e., the neat / undiluted solution) mentioned above.

[0110] A parallel study was included in which the same procedure was performed in the absence of virus to determine any residual cytotoxic effects of the formulation on the assay cells.

[0111] result The results of studies of the antiviral activity of 1% LTX-109 and 0.1% LTX-109 against respiratory syncytial virus (RSV) are summarized in Table 2 (below).

[0112] After incubation with the PBS control for 1 hour, a mean of 3.13E+05 TCID50 / ml of RSV was measured.

[0113] After 1 hour incubation with 1% LTX-109, an average of 1.58E+02 TCID50 / ml was measured, corresponding to a greater than 3 log or 99.9% reduction in infectivity compared to the PBS control.

[0114] After 1 hour incubation with 0.1% LTX-109, an average of 8.76E+01 TCID50 / ml was measured, corresponding to a greater than 3 log or 99.9% reduction in infectivity compared to the PBS control.

[0115] After filtration, cytotoxicity was observed up to the first dilution of the 1% formulation without affecting the validity of the test. No cytotoxicity was observed for the 0.1% formulation.

[0116] [Table 2]

[0117] conclusion Based on the findings reported herein, exposure of RSV to 1% or 0.1% LTX-109 for 1 hour in vitro caused a greater than 3 log reduction in viral infectivity compared to the PBS control, which corresponds to a 99.9% reduction. These results indicate that LTX-109 has excellent antiviral activity against RSV, an enveloped virus.

[0118] Example 4 Antiviral activity of LTX-109 against SARS-CoV-2 virus the aim The aim of this study was to test the antiviral activity of LTX-109 against the SARS-CoV-2 virus.

[0119] method To test whether 1% LTX-109 (w / v) has antiviral activity against SARS-CoV-2, 5 × 10 6 Infectious units of SARS-CoV2 (40 μl) were incubated with 4 volumes of 1% LTX-109 dissolved in PBS (160 μl) or PBS (phosphate buffered saline) control. Experiments were performed in triplicate.

[0120] After 1 hour, the incubation was stopped by adding excess cold medium and the formulation was physically separated from the virus through a filter to reduce cytotoxicity in the assay cells. Some turbidity during the preparation was observed for the 1% formulation, but no precipitation. Infectious virus was quantified via serial dilution (a series of 10-fold dilutions) on a monolayer of Vero cells in a microtiter plate (Vero cells are mammalian cells that can display cytopathic effects (CPE) upon viral infection). The starting solution for the serial dilution (i.e., the undiluted (or undiluted) solution) was obtained by resuspending the virus, separated via the filtration step, in 1 ml of medium. For each dilution of virus in the dilution series, eight wells of the microtiter plate were tested (i.e., each dilution of virus was applied to eight separate wells, each well containing a Vero cell monolayer). Appropriate controls were also performed. Five days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the wells at a given dilution) displayed virus-induced cytopathic effects (TCID50). TCID50 (TCID50 / ml) assay (tissue culture infectious dose 50 assay) is a type of end-point dilution assay that is well known in the art and routinely used to quantitatively measure virus titer. TCID50 / ml provides a measure of infectious units of virus / ml. " / ml" refers to / ml of the starting solution (i.e., the neat / undiluted solution) mentioned above.

[0121] A parallel study was included in which the same procedure was performed in the absence of virus to determine any residual cytotoxic effects of the formulation on the assay cells.

[0122] result The results of the studies on the antiviral activity of 1% LTX-109 against the SARS-CoV-2 virus are summarized in Table 3 (below).

[0123] After 1 hour of incubation with the PBS control, a mean of 4.27E+06 TCID50 / ml of SARS-CoV-2 was measured.

[0124] After 1 hour incubation with 1% LTX-109, an average of 1.99E+02 TCID50 / ml was measured, corresponding to a greater than 4 log or 99.99% reduction in infectivity compared to the PBS control.

[0125] After filtration, cytotoxicity was observed up to the first dilution of the 1% formulation without affecting the validity of the study.

[0126] [Table 3]

[0127] conclusion Based on the findings reported here, exposure of SARS-CoV-2 to 1% LTX-109 for 1 h in vitro caused a greater than 4 log decrease in viral infectivity compared to the PBS control, which corresponds to a 99.9% reduction. These results indicate that LTX-109 has superior antiviral activity against SARS-CoV-2, an enveloped virus.

[0128] Example 5 Antiviral activity of LTX-12 against SARS-CoV-2 virus the aim The aim of this study was to test the antiviral activity of LTX-12 against the SARS-CoV-2 virus.

[0129] method The SARS-CoV-2 isolate used was from BEI Resources: SARS-CoV-2 isolate England / 02 / 2020 (BEI Resources catalogue number (NR52359)).

[0130] To test whether LTX-12 has antiviral activity against SARS-CoV-2, 7 × 10 5 Infectious units of SARS-CoV-2 (40 μl) were incubated with 4 volumes of 1% LTX-12 (w / v) dissolved in PBS (160 μl) or PBS (phosphate buffered saline) negative control. As a positive control, a buffer containing 0.2% Triton in PBS was tested in parallel. Each sample and the PBS control were tested in triplicate.

[0131] After 1 hour at room temperature (RT), the incubation was stopped by adding excess cold assay medium (5 ml) and the formulation was physically separated from the virus via a filter (Sartorius Vivaspin 6, 100000 MWCO, PES (Sartorius, VS0642)) to reduce cytotoxicity in the assay cells. The assay medium was M199 medium (Gibco, 41150087) supplemented with 0.4% BSA (Gibco 15260037) and 1X p / s (Gibco 15070063).

[0132] Serial dilutions (series of 10-fold dilutions, 10 -1 From 10 -8 Infectious virus was quantified via ELISA using Vero cells, which are mammalian cells (African green monkey epithelial cells) that can display cytopathic effects (CPE) upon viral infection. The starting solution (i.e., undiluted (or undiluted) solution) for making the serial dilutions was prepared by resuspending the virus separated via a filtration step in 1 ml of assay medium and then serial dilutions (10 to 10) of the starting solution. -1 From 10 -8) was prepared by preparing a series of dilutions of the virus (10 -1 From 10 -8 For each dilution, eight wells of a microtiter plate were tested (i.e., each dilution of virus was applied to eight separate wells, each well containing a Vero cell monolayer). Appropriate controls were also performed. Four days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the wells at a given dilution) displayed virus-induced cytopathic effects (TCID50) using the method of Reed and Muench (LJ Reed and H. Muench, American Journal of Epidemiology, Vol. 27, No. 3, 1938, pp. 493-497). The TCID50 (TCID50 / ml) assay (tissue culture infectious dose 50 assay) is a type of end-point dilution assay that is well known in the art and routinely used to quantitatively measure virus titers. TCID50 / ml provides a measure of infectious units of virus / ml. " / ml" refers to / ml of the starting solution (i.e., neat / undiluted solution) referred to above.

[0133] A parallel study was included in which the same procedure was performed in the absence of virus to determine any residual cytotoxic effects of LTX-12 on the assay cells.

[0134] result The results of the studies on the antiviral activity of LTX-12 against the SARS-CoV-2 virus are summarized in Table 4 (below).

[0135] After 1 hour of incubation with the PBS control, a mean of 6.86E+05 TCID50 / ml of SARS-CoV-2 was measured.

[0136] After 1 hour incubation with LTX-12, an average of 1.58E+02 TCID50 / ml was measured, corresponding to a greater than 3 log or 99.977% reduction in infectivity compared to the PBS control.

[0137] After 1 hour incubation with Triton-based lysis buffer (positive control), 2.80E+01 TCID50 / ml was measured, corresponding to more than 4 logs or a 99.996% reduction in infectivity compared to the PBS control.

[0138] In the parallel studies mentioned above, no significant cytotoxicity was observed against Vero cells (assay cells).

[0139] [Table 4]

[0140] conclusion Based on the findings reported here, exposure of SARS-CoV-2 to LTX-12 for 1 h in vitro caused a reduction in SARS-CoV-2 infectivity of approximately 3.6 logs compared to the PBS control, which corresponds to at least a 99.9% reduction. These results indicate that LTX-12 has excellent antiviral activity against SARS-CoV-2, an enveloped virus.

[0141] Triton as a positive control provides a benchmark and validates the assay: enveloped viruses are known to be sensitive to Triton, and although the effect of Triton (positive control) slightly exceeds that of LTX-12, the test peptide (LTX-12) still performs well in comparison.

[0142] Cytotoxicity studies indicate that any residual peptides that may be associated with the virus after the filtration step are not responsible for the activity seen in the TCID50 assay.

[0143] Example 6 Antiviral activity of LTX-7 against influenza A virus the aim The aim of this study was to test the antiviral activity of LTX-7 against influenza A.

[0144] method The influenza A strain used was the A / WSN / 33(H1N1) strain.

[0145] To test whether LTX-7 has antiviral activity against influenza A, 5 × 10 5 Infectious units of influenza A (40 μl) were incubated with 4 volumes of 1% LTX-7 (w / v) dissolved in PBS or a PBS (phosphate buffered saline) negative control. As a positive control, a buffer containing 0.2% Triton X-100 in PBS was run in parallel. Each sample and the PBS control were run in triplicate.

[0146] After 1 hour at room temperature (RT), the incubation was stopped by adding excess cold assay medium (5 ml) and the formulation was physically separated from the virus via a filter (Sartorius Vivaspin 6, 100,000 MWCO, PES (Sartorius, VS0642)) to reduce cytotoxicity in the assay cells. The assay medium was DMEM (Gibco 61965-026) supplemented with 0.1% FBS (Gibco 10500-064), 20 mM Hepes (Gibco 15630-056), 0.3% BSA Fraction V (Gibco 15260037) and 1X p / s (Gibco 15070063).

[0147] Serial dilutions (series of 10-fold dilutions, 10 0 From 10 -7 Infectious virus was quantified via incubation with 10% ethanol at 4°C for 1 h (MDCK-II cells are mammalian cells (Madin-Darby canine kidney cells) that can display cytopathic effects (CPE) upon viral infection). Starting solutions for serial dilutions (i.e., undiluted or 10% solution) were used. 0 The virus was isolated through a filtration step and resuspended in 1 ml of assay medium. Each dilution (10 0From 10 -7 For each dilution, eight wells of a microtiter plate were tested (i.e., each dilution of virus was applied to eight separate wells, each well containing a monolayer of MDCK-II cells). Appropriate controls were also performed. Four days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the wells at a given dilution) displayed virus-induced cytopathic effects (TCID50) using the method of Reed and Muench (LJ Reed and H. Muench, American Journal of Epidemiology, Vol. 27, No. 3, 1938, pp. 493-497). The TCID50 (TCID50 / ml) assay (tissue culture infectious dose 50 assay) is a type of end-point dilution assay that is well known in the art and routinely used to quantitatively measure virus titers. TCID50 / ml provides a measure of infectious units of virus / ml. " / ml" refers to / ml of the starting solution referred to above (i.e., neat / undiluted solution).

[0148] A parallel study was included in which the same procedure was performed in the absence of virus to determine any residual cytotoxic effects of LTX-7 on the assay cells.

[0149] result The results of testing the antiviral activity of LTX-7 against influenza A virus (IAV) are summarized in Table 5 (below).

[0150] After 1 hour of incubation with the PBS control, a mean of 1.33E+06 TCID50 / ml of influenza A was measured.

[0151] After 1 hour incubation with LTX-7, an average of 1.58E+01 TCID50 / ml was measured, corresponding to a greater than 4 log or 99.99% reduction in infectivity compared to the PBS control.

[0152] After 1 hour incubation with Triton X100 lysis buffer (positive control), 1.58E+01 TCID50 / ml was measured, corresponding to a >4 log or >99.99% reduction in infectivity compared to the PBS control.

[0153] After filtration, cytotoxicity towards MDCK-II cells was observed (only) with undiluted application of LTX-7 formulation, Triton X-100 (positive control) and PBS (no virus) but did not affect the validity of the study.

[0154] [Table 5]

[0155] conclusion Based on the findings reported here, exposure of influenza A to LTX-7 for 1 hour in vitro caused a 4.75 log reduction in influenza A infectivity compared to the PBS control, which corresponds to at least a 99.99% reduction. These results indicate that LTX-7 has superior antiviral activity against influenza A (an enveloped virus).

[0156] Triton X-100 as a positive control provides a benchmark and validates the assay. Enveloped viruses are known to be sensitive to Triton X-100. LTX-7 performs well as a positive control in this study.

[0157] Cytotoxicity studies indicate that direct application of LTX-7 to MDCK-II cells is cytotoxic only before any serial dilutions are performed (i.e., in the neat formulation), and therefore any residual peptides that may be associated with the virus after the filtration step are not responsible for the activity seen in the TCID50 assay.

Claims

1. A pharmaceutical composition for the treatment of an enveloped virus infection, comprising a compound represented by the formula (II) AA1-AA2-AA1-XYZ (II) [In the formula, AA 1 is a cationic amino acid; AA 2 is an amino acid having a lipophilic R group, said R group having 14 to 27 non-hydrogen atoms; X is a branched or unbranched C 1 ~C 10 an N atom optionally substituted by an alkyl or aryl group, said group optionally incorporating up to two heteroatoms selected from N, O and S; Y is -R a -R b -, -R a -R b -R b -and-R b -R b -R a -, wherein R a is C, O, S or N, R b is C and R a and R b Each of the 1 ~C 4 may be substituted or unsubstituted by alkyl groups, Z is a group containing from 1 to 3 cyclic groups of 5 or 6 non-hydrogen atoms each, two or more of which may be fused, one or more of which may be substituted, the Z moiety incorporating up to 15 non-hydrogen atoms, wherein The bond between Y and Z is the R of Y. a or R b and a non-hydrogen atom of one of said cyclic groups of Z. A pharmaceutical composition comprising a compound of formula (I).

2. 2. The pharmaceutical composition according to claim 1, wherein the cationic amino acid is lysine or arginine, preferably arginine.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein the lipophilic R group contains two or more cyclic groups which may be fused or connected.

4. 3. The pharmaceutical composition of claim 1 or 2, wherein X is unsubstituted.

5. Y is -R a -R b 3. The pharmaceutical composition according to claim 1 or 2, wherein Y is - and is unsubstituted, preferably Y is -CH2-CH2-.

6. 3. The pharmaceutical composition of claim 1 or 2, wherein Z is phenyl.

7. A pharmaceutical composition described in claim 1 or 2, wherein AA 2 is selected from tributyltryptophan (Tbt) or a biphenylalanine derivative selected from Phe(4-(2-naphthyl)), Phe(4-(1-naphthyl)), Bip(4-n-Bu), Bip(4-Ph) and Bip(4-T-Bu), preferably AA 2 is Tbt.

8. -XYZ together -NHCH 2 CH 2 The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is Ph.

9. The compound of formula (II) having the structural formula: 【Chemistry 1】 3. The pharmaceutical composition according to claim 1 or 2, comprising:

10. The pharmaceutical composition according to claim 1 or 2, wherein the enveloped virus infection is a respiratory tract infection.

11. 3. The pharmaceutical composition of claim 1 or 2, wherein the enveloped virus is a coronavirus, an orthopneumovirus or an orthomyxovirus.

12. The pharmaceutical composition of claim 1 or 2, wherein the enveloped virus is SARS-CoV-2, respiratory syncytial virus (RSV) or influenza virus.

13. The pharmaceutical composition of claim 1 or 2, further comprising a diluent, carrier and / or excipient.

14. The pharmaceutical composition according to claim 1 or 2, wherein the treatment is a therapeutic treatment.

15. The pharmaceutical composition according to claim 1 or 2, wherein the treatment is a prophylactic treatment.

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