Modified tripeptides for use in the treatment of non-enveloped viral infections - Patents.com

Abstract

The invention described herein relates to modified tripeptides, in particular LTX-109, for use in the treatment of non-enveloped viral infections.

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 non-enveloped viral 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, so the capsid is the outermost part. Non-enveloped viruses are sometimes called "naked" viruses.

[0003] Viral infections represent a significant medical problem. For example, at least 50% of people presenting with cold symptoms have an underlying rhinovirus infection, making rhinoviruses one of the most common and significant non-enveloped viruses in humans. There is an enormous societal cost associated with colds in terms of missed school and work. Rhinovirus infections are also responsible for more severe conditions such as childhood otitis media and exacerbations of childhood asthma, as well as sinusitis. Other diseases caused by non-enveloped viruses include polio, aseptic meningitis, papillomas (warts), and acute infantile diarrhea (winter diarrhea; rotavirus).

[0004] Non-enveloped viruses lack the fragile lipid envelope and may therefore be more resistant to some disinfectants and other measures (pH and temperature) that may be used to inhibit viruses, which may also explain the smaller number of agents available to treat non-enveloped virus infections than enveloped virus infections.

[0005] Nevertheless, there is great interest in effective anti-non-enveloped virus agents. For example, 25-hydroxycholesterol (25HC) has been shown to have significant antiviral activity against three pathogenic non-enveloped viruses, namely human papillomavirus-16 (HPV-16), human rotavirus (HRoV) and human rhinovirus (HRhV). Interferon-alpha has been shown to be effective against human rhinovirus infections, but side effects and the development of tolerability by volunteers have led to the abandonment of research into this treatment. Pleconaril is a drug that prevents rhinoviruses from attaching to host cells, but resistance can emerge that causes mutations in the capsid protein (VP1) to which the drug is bound, reducing efficacy. Viral resistance to antiviral agents is a significant problem in international health care. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] WO2009 / 081152A2 [Non-patent literature]

[0007] [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]

[0008] 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]

[0009] The present inventors have surprisingly found that a class of tripeptide compounds having certain C-terminal modifications exhibit superior antiviral activity against non-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 antibacterial 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-non-enveloped virus therapies.

[0010] Thus, in one aspect, the present invention provides a compound for use in treating a non-enveloped viral infection in a subject, the compound comprising the compound 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,

[0011] 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.

[0012] 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.

[0013] The compounds for use according to the present invention are preferably peptides.

[0014] 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.

[0015] 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):

[0016] 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)), Phe(4-(1-naphthyl)) and Tbt being most preferred. In some preferred embodiments, the amino acid with a lipophilic R group is tributyltryptophan (Tbt).

[0017] 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-.

[0018] In some preferred embodiments, Z is phenyl (Ph).

[0019] A further preferred group of compounds is where -XYZ together are a -NHCH2CH2Ph ​​group.

[0020] 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.

[0021] The most preferred compounds have the structural formula:

[0022] [ka]

[0023] The formula is: 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 (which may also be referred to as tributyltryptophan (Tbt)) 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. In one embodiment, one of the Arg residues in LTX-109, such as the N-terminal Arg or the C-terminal Arg, is replaced by a Lys residue. In another embodiment, both Arg residues in LTX-109 are replaced by Lys residues. In one embodiment, one of the Arg residues in LTX-109, such as the N-terminal Arg or the C-terminal Arg, is replaced by a His residue. In another embodiment, both Arg residues in LTX-109 are replaced by His residues.

[0024] Other cationic residues in place of Arg include suitable non-genetically encoded and modified amino acids, including 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.

[0025] Also highly preferred are analogs incorporating alternative C-terminal capping groups as defined above.

[0026] Further preferred compounds for use according to the present invention are

[0027] [ka]

[0028] 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.

[0029] Further preferred compounds for use according to the present invention are

[0030] [ka]

[0031] 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.

[0032] In some preferred embodiments, the compound for use according to the present invention is LTX-109 or LTX-7. The compound LTX-109 is the most preferred compound for use according to the present invention.

[0033] The compounds for use in the present invention are preferably peptides.

[0034] 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).

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] 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 will be present on the α-amino group of each new residue added and will need to be selectively removed prior to the next coupling step.

[0044] For example, carboxyl protecting groups which may be used include easily cleaved ester groups, such as benzyl (Bzl), p-nitrobenzyl (ONb), pentachlorophenyl (OPClP), pentafluorophenyl (OPfp) or t-butyl (OtBu) groups, and methyl groups which are linked to a coupling group on a solid support, such as polystyrene.

[0045] Thiol protecting groups include p-methoxybenzyl (Mob), trityl (Trt) and acetamidomethyl (Acm).

[0046] 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.

[0047] 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.

[0048] Compounds for use according to the present invention (eg, LTX-109) may be synthesized as described in WO2009 / 081152A2.

[0049] The compounds (e.g., peptides) for use according to the invention exhibit activity against non-enveloped viruses, in other words, the compounds for use according to the invention exhibit anti-non-enveloped virus activity.

[0050] The compounds used in the present invention typically exhibit activity against non-enveloped viruses (anti-non-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). Those skilled in the art are familiar with suitable in vitro assays, such as suitable end-point dilution assays (e.g., TCID50 assays). A preferred TCID50 assay is described in the Examples section of this specification.

[0051] Without wishing to be bound by theory, it is believed that the compounds used in the present invention do not target specific proteins, but instead have a more general mechanism of action.In particular, it is believed that there is an electrostatic mechanism, whereby the positively charged compounds of the present invention are attracted to the negatively charged regions on the surface of non-enveloped viruses.This is advantageous both in terms of the breadth of viruses that can be treated and in terms of avoiding the development of resistance through specific mutations in viral proteins.

[0052] As indicated above, the present invention provides a compound as defined elsewhere herein for use in treating a non-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 a non-enveloped virus.

[0053] "Non-enveloped viruses" are viruses that lack a lipid layer (or lipid membrane). Thus, non-enveloped viruses have a capsid (viral protein capsid) as their outermost layer. The capsid shell surrounds the viral genome.

[0054] In accordance with the present invention, preferred target viruses are icosahedral in their capsid shape (these viruses are known as icosahedral viruses). Although there are a variety of different sizes and arrangements of capsid proteins, all of these icosahedral viruses have 20 triangular faces made up of capsid proteins, forming an approximately spherical shape.

[0055] Any non-enveloped viral infection can be treated according to the present invention. Typically and preferably, the non-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 non-enveloped virus according to the present invention is a mammalian pathogen, preferably a human pathogen.

[0056] In some embodiments, the non-enveloped virus is the causative agent of respiratory tract infection, also known as respiratory virus. The respiratory tract infection may be an infection of the upper and / or lower respiratory tract. The upper respiratory tract infection is the preferred target for treatment according to the present invention.

[0057] The non-enveloped virus may be a DNA virus or an RNA virus. In some preferred embodiments, the non-enveloped virus is an RNA virus (e.g., a single-stranded (ss) RNA non-enveloped virus).

[0058] Preferred target viruses are members of the Picornaviridae, Calciviridae, Parvoviridae, Papovaviridae, Papillomaviridae and Reoviridae families, with picornaviruses being particularly preferred.

[0059] Among the picornaviruses, the Enterovirus genus is a preferred target. Structurally, all enteroviruses are small, between 15 and 30 nm. The capsid contains a single-stranded positive stranded RNA (+ssRNA) of approximately 7400 nucleotides in length. Instead of having an AUG-containing cap, the genome has an internal ribosome entry site (IRES) allowing mRNA translation. Within the Enterovirus genus, enteroviruses, coxsackieviruses, rhinoviruses, echoviruses and polioviruses are found, which are preferred viral targets according to the present invention. They are the causative agents for a wide variety of diseases ranging from the common cold (sometimes also called viral rhinitis) to acute poliomyelitis and aseptic meningitis. In humans, they are one of the most common infectious agents worldwide. In some embodiments, the non-enveloped virus may be a virus of one of the following species: Enteroviruses A-D and Rhinoviruses A-C. Rhinoviruses (all serotypes) are particularly preferred.

[0060] In some embodiments, the virus from the Enterovirus genus can be Enterovirus C (sometimes referred to as Enterovirus species C or Enterovirus type C) or Enterovirus D (sometimes referred to as Enterovirus species D or Enterovirus type D). In some embodiments, the Enterovirus C can be Enterovirus C104 (EV-C104), Enterovirus C105 (EV-C105), Enterovirus C109 (EV-C109), Enterovirus C117 (EV-C117) or Enterovirus C118 (EV-C118). In some embodiments, the Enterovirus D can be Enterovirus D68 (EV-D68). Enterovirus C can cause, for example, a cold (viral rhinitis) and / or pneumonia. Enterovirus D (e.g., Enterovirus D68) can cause, for example, pneumonia.

[0061] In some embodiments, the virus from the Enterovirus genus may be a Coxsackievirus. The Coxsackievirus may be a Group A Coxsackievirus (e.g., Coxsackievirus A21, also known as CV-A21) or a Group B Coxsackievirus. The Coxsackievirus may, for example, cause an upper respiratory tract infection, such as the common cold (viral rhinitis).

[0062] In some embodiments, the virus from the Enterovirus genus may be an echovirus. Echoviruses can cause, for example, upper respiratory tract infections, such as viral rhinitis. Upper respiratory tract infections caused by echoviruses can occur especially in children.

[0063] As indicated above, in some embodiments, rhinoviruses are preferred viruses according to the present invention. In some embodiments, the rhinovirus may be rhinovirus A (e.g., human rhinovirus 60), rhinovirus B (e.g., human rhinovirus 14) and / or rhinovirus C.

[0064] In some embodiments, the virus from the Papillomaviridae family is a Human Papillomavirus (HPV). Human Papillomaviruses can cause, for example, skin or mucosal growths (warts). In some embodiments, the virus from the Caliciviridae family is a Norovirus. Noroviruses can cause, for example, gastroenteritis. Norovirus infections are typically characterized by non-bloody diarrhea, vomiting, stomach pain, fever and / or headache. 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 a non-enveloped virus infection. The embodiments of the other aspects of the invention described herein apply mutatis mutandis to this aspect of the invention.

[0065] Diseases or conditions to be treated include infections of the upper or lower respiratory tract, including the common cold (a term used to refer to a collection of symptoms including / selected from: stuffy or runny nose, sore throat, sneezing, coughing, muscle pain, headache, sinus pain, and fatigue), otitis media, sinusitis, pneumonia, bronchopneumonia, aseptic meningitis, polio, epidemic myalgia, hand, foot and mouth disease, myocarditis, pericarditis, pneumonitis, and cerebella ataxia. The common cold and its symptoms are particularly preferred target conditions (patients will typically present with at least two or three cold symptoms).

[0066] In some embodiments, the compounds (or formulations or compositions) according to the invention are for use in treating a subject with an upper or lower respiratory tract infection (e.g., the common cold), e.g., reducing the severity and / or frequency of symptoms. In such embodiments, the upper or lower respiratory tract infection may be caused by a non-enveloped viral infection (e.g., rhinovirus). In some embodiments, the invention is for use in treating, e.g., reducing the severity and / or frequency of, stuffy or runny nose, sore throat, sneezing, coughing, muscle aches, headaches, sinus pain, and / or fatigue.

[0067] Other diseases or conditions that may be treated include HPV infection. Thus, warts (e.g., cutaneous warts or genital warts) are examples of other conditions that may be treated in accordance with the present invention.

[0068] In some embodiments, the compounds (or formulations or compositions) according to the invention are for use in treating a subject with an HPV infection, e.g., reducing the severity and / or frequency of symptoms. In some embodiments, the invention is for use in treating skin or mucosal growths (warts), e.g., reducing the severity and / or frequency thereof.

[0069] Other diseases or conditions that may be treated include Norovirus infection. Thus, gastroenteritis caused by Norovirus is an example of a condition that may be treated according to the present invention.

[0070] In some embodiments, a compound (or formulation or composition) according to the invention is for use in treating a subject with a Norovirus infection, e.g., reducing the severity and / or frequency of symptoms. In some embodiments, the invention is for use in treating, e.g., reducing the severity and / or frequency of diarrhea (e.g., non-bloody diarrhea), vomiting, stomach pain, fever, and / or headache.

[0071] 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 non-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 pharma-ceutically acceptable diluent, carrier and / or excipient.

[0072] 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.

[0073] The compounds (or formulations or compositions) for use according to the present invention may be administered orally, nasally, parenterally, intravenously, topically or rectally.

[0074] In some embodiments, the compounds (or formulations or compositions) of the present invention are for administration to the upper or lower respiratory tract. For example, compositions and formulations for use according to the present invention may be administered using, for example, a microcatheter (e.g., endoscope and microcatheter), an aerosolizer, a powder dispenser, a nebulizer, or an inhaler. Optionally, in some embodiments, the compounds (or formulations or compositions) are administered as finely divided powders or liquid aerosols.

[0075] In some embodiments, the compound (or formulation or composition) of the present invention is for nasal administration. For example, the compositions and formulations for use according to certain embodiments of the present invention can be administered using a nasal applicator. Optionally, in some embodiments, the compound (or formulation or composition) is administered to the subject in a nasal formulation (e.g., as a finely divided powder or liquid solution).

[0076] Compositions for use according to certain embodiments of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art.

[0077] 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.

[0078] As used herein, the term "pharmaceutical" includes veterinary applications of the present invention.

[0079] The compounds defined herein may be presented in conventional pharmaceutical dosage forms, such as tablets, coated tablets, solutions, emulsions, liposomes, powders, capsules, suppositories or sustained release forms.

[0080] Conventional pharmaceutical excipients and normal manufacturing methods may be used for preparing these forms.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] Treatment according to the invention may involve co-administration with one or more additional active agents used in the treatment or prevention of non-enveloped viral 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.

[0088] The term "treatment" or "therapy" as used herein includes therapeutic and prophylactic (or protective) therapy. Thus, the compounds for use according to the invention may be for therapeutic or protective use. A "prophylactic (or protective) treatment" is a treatment administered to a subject who does not (or has not yet) exhibit signs or symptoms of a disease, or who exhibits only early signs or symptoms thereof, such that the treatment is administered with the aim of preventing or reducing the risk of developing a disease and / or symptoms associated with the disease. A protective treatment functions as a treatment that inhibits or reduces the further development or enhancement of a disease and / or its associated symptoms. A "therapeutic treatment" is a treatment administered to a subject who displays symptoms or signs of a disease, where the treatment is administered to the subject with the aim of reducing or eliminating those signs or symptoms, e.g. reducing the severity and / or frequency of the symptoms, or slowing or halting disease progression.

[0089] Viewed alternatively, the present invention provides a method of treating a non-enveloped viral 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.

[0090] The present invention also provides a method of treating a disease or condition caused by (or characterised by) a non-enveloped viral 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.

[0091] 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 target non-enveloped virus, or to prevent or reduce their reproduction rate, or to otherwise reduce their harmful effects on the body. Administration may be protective. Such an effective amount may be administered once, i.e., in a single dose, or several times, i.e., in a series of doses, for example, over the course of several days, weeks, or months.

[0092] 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 a non-enveloped viral 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.

[0093] 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 a disease or condition caused by (or characterised by) a non-enveloped viral 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.

[0094] Viewed yet another way, the present invention provides the use of a compound as defined herein for the treatment of a non-enveloped viral 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.

[0095] 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) a non-enveloped viral 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.

[0096] 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.

[0097] In some embodiments, the subject according to the present invention is a subject with a non-enveloped viral infection. In some embodiments, the subject according to the present invention is a subject suspected of having a non-enveloped viral infection. In some embodiments, the subject according to the present invention may be a subject at risk of developing (or at risk of contracting) a non-enveloped viral infection.

[0098] In some embodiments, the subject according to the present invention is a subject with a disease or condition caused by a non-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 a non-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 a non-enveloped virus infection.

[0099] The invention also provides kits comprising one or more of the compounds or compositions according to the invention for use in the methods and uses described herein. Preferably, the kits include instructions for use in treating a non-enveloped viral infection as described herein.

[0100] 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.

[0101] 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.

[0102] The invention will now be further described with reference to the following non-limiting examples. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS EXAMPLES

[0103] Example 1 Antiviral activity of 1% LTX-109 against rhinovirus (rhinovirus 60) the aim The aim of this study was to test the antiviral activity of 1% LTX-109 against rhinovirus.

[0104] method The rhinovirus used was from BEI Resources: Rhinovirus 60, 2268-CV37 (catalog number NR-51447).

[0105] To test whether 1% LTX-109 (w / v) has antiviral activity against rhinovirus, 2.24 × 10 5 Infectious units of rhinovirus (40 μl) were incubated with 4 volumes of 1% LTX-109 dissolved in PBS (160 μl), along with a PBS (phosphate buffered saline) control and a 0.25% SDS (sodium dodecyl sulfate) positive control. Treatments were performed in triplicate.

[0106] After 1 hour, the incubation was stopped by adding excess medium and the virus was separated by filtering the liquid through a filter (Sartorius Vivaspin 6) to reduce cytotoxicity to the assay cells. The assay medium was DMEM (Gibco 61965-026) supplemented with 2% FBS (Gibco 10500-064), 20 mM Hepes (Gibco 15630-056) and 1X p / s (Gibco 15070063).

[0107] Infectious virus was quantified via serial dilution of the filtrate (a series of 10-fold dilutions) on a monolayer of HeLa cells in a microtiter plate (HeLa cells are human cells that can exhibit cytopathic effects (CPE) upon viral infection). For each dilution of virus in the dilution series, eight wells of the microtiter plate were tested (i.e., each dilution was applied to eight separate wells, each well containing a HeLa cell monolayer). Appropriate controls were also performed. Seven days after infection of the cells, virus titers were quantified by determining the dilution at which half of the cells (half of the wells of a given dilution) displayed virus-induced cytopathic effects (TCID50). 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 is 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).

[0108] 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.

[0109] result The results of testing the antiviral activity of 1% LTX-109 against rhinovirus are summarized in Table 1 (below).

[0110] After 1 hour incubation with the PBS control, a mean of 4.3E+05 TCID50 / ml of rhinovirus was measured.

[0111] After 1 hour incubation with 1% LTX-109, an average of 4.8E+03 TCID50 / ml was measured, corresponding to a 1.9 log reduction in infectivity compared to the PBS control.

[0112] After filtration, cytotoxicity towards HeLa cells was observed (only) with neat application (without virus) of the 1% LTX-109 formulation, but this did not affect the validity of the study.

[0113] [Table 1]

[0114] conclusion Based on the findings reported here, exposure of rhinovirus to 1% LTX-109 for 1 hour in vitro caused a 1.9 log decrease in viral infectivity compared to the PBS control, which corresponds to approximately a 99% reduction compared to the PBS control. These results indicate that LTX-109 has excellent antiviral activity against rhinovirus (a non-enveloped virus).

[0115] SDS as a positive control provides a benchmark and validates the suitability of the assay. Non-enveloped viruses are known to be sensitive to SDS, and although the effect of SDS is slightly greater than that of LTX-109, the test peptide still performs well in comparison.

[0116] Cytotoxicity studies indicate that direct application of LTX-109 to HeLa cells is cytotoxic only before any serial dilutions are performed (i.e., in the neat formulation). 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.

[0117] Example 2 Antiviral activity of 3% LTX-109 against rhinovirus (rhinovirus 60) the aim The aim of this study was to test the antiviral activity of 3% LTX-109 against rhinovirus.

[0118] method The rhinovirus used was from BEI Resources: Rhinovirus 60, 2268-CV37 (catalog number NR-51447).

[0119] To test whether 3% LTX-109 (w / v) has antiviral activity against rhinovirus, 9 × 10 5 Infectious units of rhinovirus (40 μl) were incubated with 4 volumes of 3% LTX-109 dissolved in PBS (160 μl), along with a PBS (phosphate buffered saline) control and a 0.25% SDS (sodium dodecyl sulfate) positive control. Treatments were performed in triplicate.

[0120] After 1 hour, the incubation was stopped by adding 50 μl of the mixture to 5 ml of medium. Assay medium was DMEM (Gibco 61965-026) supplemented with 2% FBS (Gibco 10500-064), 20 mM Hepes (Gibco 15630-056) and 1X p / s (Gibco 15070063).

[0121] Infectious virus was quantified via serial dilution (a series of 10-fold dilutions) on monolayers of HeLa cells in microtiter plates (HeLa cells are human cells that can display cytopathic effects (CPE) upon viral infection). For each dilution of virus in the dilution series, eight wells of the microtiter plate were tested (i.e., each dilution was applied to eight separate wells, each well containing a HeLa cell monolayer). Appropriate controls were also performed. Seven days after infection of the cells, virus titers were quantified by determining the dilution at which half of the cells (half of the cells at a given dilution) displayed virus-induced cytopathic effects (TCID50). 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) mentioned above.

[0122] 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.

[0123] result The results of testing the antiviral activity of 3% LTX-109 against rhinovirus are summarized in Table 2 (below).

[0124] After 1 hour of incubation with the PBS control, 1.76 × 10 4 The mean TCID50 / ml was determined.

[0125] After 1 h of incubation with 3% LTX-109, 1.99 × 10 2 The mean TCID50 / ml was determined, which corresponds to a 1.9 log reduction in infectivity compared to the PBS control.

[0126] After filtration, cytotoxicity towards HeLa cells was observed (only) in the neat application of the diluted 3% formulation (without virus), but this did not affect the validity of the study.

[0127] [Table 2]

[0128] conclusion Based on the findings reported here, exposure of rhinovirus to 3% LTX-109 for 1 hour in vitro caused a 1.9 log decrease in viral infectivity compared to the PBS control, which corresponds to approximately a 99% reduction compared to the PBS control. These results indicate that LTX-109 has excellent antiviral activity against rhinovirus (a non-enveloped virus).

[0129] SDS as a positive control provides a benchmark and validates the suitability of the assay. Non-enveloped viruses are known to be sensitive to SDS, and although the effect of SDS is slightly greater than that of LTX-109, the test peptide still performs well in comparison.

[0130] Cytotoxicity studies indicate that direct application of LTX-109 to HeLa cells is cytotoxic only before any serial dilutions are performed (i.e., in the neat liquid), and therefore any residual peptides that may be associated with the virus are not responsible for the activity seen in the TCID50 assay.

[0131] Example 3 Antiviral activity of LTX-7 against rhinovirus (rhinovirus 60) the aim The aim of this study was to test the antiviral activity of LTX-7 against rhinovirus.

[0132] method The rhinovirus used was from BEI Resources: Rhinovirus (HRV-A60), strain: 2268-CV37 (BEI Resources catalog number NR-51447).

[0133] To test whether LTX-7 has antiviral activity against rhinovirus, 5 × 10 5 Infectious units of rhinovirus (40 μl) were incubated with 4 volumes of 1% LTX-7 (w / v) dissolved in PBS (160 μl) or PBS (phosphate buffered saline) negative control. As a positive control, a buffer containing 0.25% SDS (sodium dodecyl sulfate) in PBS was run in parallel. Each sample and the PBS control were run in triplicate.

[0134] 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. Assay medium was DMEM (Gibco 61965-026) supplemented with 2% FBS (Gibco 10500-064), 20 mM Hepes (Gibco 15630-056), and 1X p / s (Gibco 15070063).

[0135] Serial dilutions (series of 10-fold dilutions, 10 0 From 10 -7 Infectious virus was quantified via ELISA (HeLaM cells are human cells that can display cytopathic effect (CPE) upon viral infection). Starting solutions for serial dilutions (i.e., undiluted or 10 0 The virus was isolated through a filtration step and resuspended in 1 ml of assay medium. Each dilution (10 0 From 10 -7For each dilution, eight wells of a microtiter plate were tested (i.e., each dilution was applied to eight separate wells, each well containing a HeLaM cell monolayer). Appropriate controls were also performed. Seven days after infection of the cells, the virus titer was quantified by determining the dilution at which half of the cells (half of the cells 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.

[0136] 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.

[0137] result The results of studies on the antiviral activity of LTX-7 against rhinoviruses are summarized in Table 3 (below).

[0138] After incubation with the PBS control for 1 hour, a mean of 9.17E+04 TCID50 / ml of rhinovirus was measured.

[0139] After 1 hour incubation with LTX-7, an average of 3.86E+03 TCID50 / ml was measured, corresponding to a 1.33 log or at least a 90% reduction in infectivity compared to the PBS control.

[0140] After 1 hour incubation with SDS (positive control), an average of 1.58E+01 TCID50 / ml was measured, which corresponds to a 3.67 log or approximately 99.9% reduction in infectivity compared to the PBS control.

[0141] After filtration, cytotoxicity towards HeLaM cells was observed (only) with neat application of LTX-7 formulation and SDS (virus free), but did not affect the validity of the study.

[0142] [Table 3]

[0143] conclusion Based on the findings reported here, exposure of rhinovirus to LTX-7 for 1 hour in vitro caused a 1.33 log decrease in rhinovirus infectivity compared to the PBS control, which corresponds to at least a 90% reduction. These results indicate that LTX-7 has superior antiviral activity against rhinoviruses (non-enveloped viruses).

[0144] SDS as a positive control provides a benchmark and validates the suitability of the assay. Non-enveloped viruses are known to be sensitive to SDS, and although the effect of SDS exceeds that of LTX-7, the test peptide (LTX-7) still performs well in comparison.

[0145] Cytotoxicity studies indicate that direct application of LTX-7 to HeLaM 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.

[0146] Example 4 Antiviral activity of 1% LTX-109 against rhinovirus (human rhinovirus 14). the aim The aim of this study was to test the antiviral activity of 1% LTX-109 against human rhinovirus-14.

[0147] method The virus used was from ATCC: Human rhinovirus 14, strain 1059 (ATCC VR-284, lot number 70049530).

[0148] To test whether 1% LTX-109 has virucidal activity against human rhinovirus 14, 40 μl of 4 × 10 7 Infectious units of human rhinovirus 14 were incubated with 4 volumes (160 μl) of 1% LTX-109 or PBS negative control. As a positive control, a buffer containing 2.5% glutaraldehyde in PBS was tested in parallel. Both LTX-109 and the negative control were tested in triplicate. The positive control was tested in a single replicate.

[0149] 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) to reduce cytotoxicity to the assay cells. Assay medium was DMEM (Gibco 10566016) supplemented with 2% FBS (Gibco 10500064), 20 mM HEPES (Gibco 15630056) and 1X PenStrep (Gibco 15070063).

[0150] Concentrated virus was resuspended in 1 ml of assay medium and serially diluted (10 -1 From 10 -8) for each dilution of virus in the dilution series, four wells of a microtiter plate were tested (i.e., each dilution was applied to four separate wells, each well containing a HeLaM cell monolayer). Specifically, on a 96-well plate, 225 μl of medium was placed in each well of four columns (32 wells in total). In the four wells in the top row (row A), 25 μl of resuspended virus was added and mixed. Then, with a multichannel pipette, 25 μl of medium was transferred from four wells in row A to four wells in the adjacent row B, and then to an equal row H, taking row B to row C, to obtain serial dilutions (10 -1 From 10 -8 ) was prepared. 200 μl from each well was then added to a separate well containing a monolayer of HeLa M cells. Appropriate controls were also performed. Three days after infection, virus titers were quantified by determining the dilution at which half of the cells displayed virus-induced cytopathic effects (TCID50) using the method of Reed and Muench (L. J. Reed and H. Muench, American Journal of Epidemiology, Vol. 27, No. 3, 1938, pp. 493-497).

[0151] 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-109 on the assay cells.

[0152] result The results of the antiviral activity of 1% LTX-109 against rhinovirus 14 are summarized in Table 4 (below).

[0153] After 1 hour incubation with the PBS control, a mean of 2.04E+07 TCID50 / ml of rhinovirus 14 was measured.

[0154] After 1 hour incubation with LTX-109, an average of 1.80E+06 TCID50 / ml was measured, corresponding to a 1.05 log reduction in infectivity, corresponding to a 91.2% reduction in rhinovirus 14 infectivity compared to the PBS control.

[0155] After 1 hour incubation with 2.5% glutaraldehyde in PBS (positive control), an average of 3.75E+01 TCID50 / ml was measured, which corresponds to a 5.74 log or approximately 99.9% reduction in infectivity compared to the PBS control.

[0156] After filtration and resuspension, -1 Cytotoxicity was observed for 1% LTX-109 when added to assay cells at dilutions. No significant cytotoxicity was observed at larger dilutions. No significant cytotoxicity was observed for PBS.

[0157] [Table 4]

[0158] conclusion Based on the findings reported here and under the conditions tested, exposure of human rhinovirus 14 to 1% LTX-109 for 1 hour caused a 1.05 log reduction in human rhinovirus 14 infectivity, which corresponds to a 91.2% reduction.

[0159] Glutaraldehyde as a positive control provides a benchmark and validates the suitability of the assay. Non-enveloped viruses are known to be sensitive to glutaraldehyde, and although the effect of the positive control exceeds that of LTX-109, the test peptide (LTX-109) performs well in comparison.

[0160] After filtration and resuspension, -1 Cytotoxicity was observed only for 1% LTX-109 when added to assay cells at dilutions. No significant cytotoxicity was observed at larger dilutions.

[0161] These results indicate that LTX-109 has superior antiviral activity against human rhinovirus 14, a non-enveloped virus.

Claims

1. A pharmaceutical composition for the treatment of a non-enveloped viral infection, said pharmaceutical composition comprising a compound of formula (II): AA 1 -AA 2 -AA 1 -XYZ (II) wherein AA 1 is a cationic amino acid; AA 2 is an amino acid with a lipophilic R group, the 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, 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, 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 1 to 3 cyclic groups of 5 or 6 non-hydrogen atoms each, two or more of the cyclic groups may be fused, one or more of the cyclic groups may be substituted, and the Z moiety incorporates 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 the cyclic groups of Z. A pharmaceutical composition comprising:

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

3. 3. The pharmaceutical composition of claim 1, 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, 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 compound is Ph.

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

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

11. The pharmaceutical composition according to claim 1 or 2, wherein the non-enveloped viral infection is a respiratory tract infection.

12. 3. The pharmaceutical composition according to claim 1, wherein the non-enveloped virus is a virus of the Picornaviridae family, preferably the non-enveloped virus is rhinovirus A, rhinovirus B or rhinovirus C.

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

14. 3. The pharmaceutical composition of claim 1 or 2, wherein the treatment is administered using a microcatheter, an aerosolizer, a powder dispenser, a nebulizer, an inhaler and / or a nasal applicator.

15. 3. The pharmaceutical composition of claim 1 or 2, wherein the treatment is a therapeutic treatment.

16. 3. The pharmaceutical composition of claim 1 or 2, wherein the treatment is a prophylactic treatment.

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