KR102393872B1 - Antiviral peptide from M2 protein and the use thereof - Google Patents

Abstract

The present invention relates to an antiviral peptide and a composition for preventing or treating a viral disease comprising the antiviral peptide as an active ingredient.
The peptides derived from M2 protein of the present invention have amphiphilic properties having both hydrophilicity and hydrophobicity, and have high selectivity for the outer membrane of the virus. can be used readily.

Description

Antiviral peptide from M2 protein and the use thereof that damage the viral envelope

The present invention relates to a peptide that inhibits outer membrane virus infection by damaging the viral envelope, and more particularly, to an M2 protein-derived peptide of the present invention and an antiviral composition comprising the same.

Antiviral efficacy refers to the ability of a virus to inhibit infection, and in particular, the general mechanism of action of antiviral peptides is known as inhibition of viral membrane protein function, inhibition of viral RNA amplification in the nucleus of infected cells, inhibition of infection through viral envelope damage, and the like. In the case of enveloped viruses, the envelope has a lipid bilayer structure, so inactivation of the envelope means loss of infectivity of the virus.

A characteristic of the peptides showing antiviral efficacy by attacking the outer membrane of the virus is that they have amphipathic properties. The amphipathic peptide refers to a peptide having both hydrophilicity (polarity, hydrophilicity) and hydrophobicity (non-polarity, hydrophobicity) within a single helix. Amphiphilic peptides coexist with hydrophilic amino acids and hydrophobic amino acids within a single helix, and acquire amphiphilicity through a biased distribution within the alpha helix structure. Through these properties, it can easily act on the lipid bilayer, which is hydrophobic on the outside and hydrophilic on the inside, and it is known that various amphiphilic peptides have antiviral efficacy.

The secondary structure of proteins, α-helix, is the most common structural motif found in proteins, and plays a key role in various biological recognition processes inside and outside cells, such as protein-DNA, protein-RNA, and protein-protein interactions. This interaction is very important in maintaining and regulating the various physiological processes of cells and living things. Accordingly, rather than using a large-sized protein as it is, research is being actively conducted to develop an alpha-helix peptide that is easy to handle and has excellent cell permeability to control or inhibit various interactions of living things.

On the other hand, M2 protein is a surface protein that plays an important role in virus proliferation, and is known to be involved in the budding process of the cell membrane in the process of proliferating and releasing the next-generation virus after the influenza virus infects the cell. Peptides derived from the cytoplasmic tail of this protein were also found to directly affect the lipid bilayer, and it has been demonstrated to induce budding in liposomes. In addition, the amphipathic peptide derived from the M2 protein induces direct changes without completely degrading or destroying the lipid bilayer membrane.

Therefore, the present inventors have been repeating research to develop an amphipathic peptide with increased selectivity for the viral outer membrane in order to solve the above problem, and the present invention derived from M2 protein that can reduce cytotoxicity by minimizing the effect on the cell membrane Viral peptides were completed.

It is an object of the present invention to provide antiviral peptides.

In addition, it is an object of the present invention to provide a composition for preventing or treating a viral disease comprising an antiviral peptide as an active ingredient.

In order to achieve the above object, the present invention provides antiviral peptides.

In addition, the present invention provides a composition for preventing or treating a viral disease comprising an antiviral peptide as an active ingredient.

The peptides derived from M2 protein of the present invention have amphiphilic properties having both hydrophilicity and hydrophobicity, and have high selectivity for the outer membrane of the virus. can be used readily.

1 is a diagram showing the sequence and helix structure of a peptide derived from M2 protein.
2 is a diagram showing the antiviral effect of peptides derived from M2 protein.
3 is a diagram showing a wide range of effects (A) and cytotoxicity (B) of peptides derived from M2 protein.
4 is a diagram showing the modification of the liposome lipid bilayer by the M2 protein-derived peptide.
5 is a diagram showing the damage and modification of the viral envelope by M2 protein-derived peptides.

Hereinafter, the present invention will be described in detail by way of embodiments of the present invention with reference to the accompanying drawings. However, the following embodiments are presented as examples of the present invention, and when it is determined that detailed descriptions of well-known techniques or configurations known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description may be omitted, and , the present invention is not limited thereby. Various modifications and applications of the present invention are possible within the scope of equivalents interpreted therefrom and the description of the claims to be described later.

In addition, the terms used in this specification are terms used to properly express the preferred embodiment of the present invention, which may vary depending on the intention of a user or operator or customs in the field to which the present invention belongs. Accordingly, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

All technical terms used in the present invention, unless otherwise defined, have the meaning as commonly understood by one of ordinary skill in the art of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention. The contents of all publications herein incorporated by reference are incorporated herein by reference.

In one aspect, the present invention provides RLX1X2KX3X4YRX5X6X7X8GLKRG, wherein X1 is S or F, X2 is S or F, X3 is S or C, X4 is I or V, and X5 is F, I, R, L or H , X6 is F or L, X7 is E or K, and X8 is H or Y.

In one embodiment, the antiviral peptide may consist of the amino acid sequence of RLFFKCIYRFFEHGLKRG (SEQ ID NO: 1).

In one aspect, the present invention relates to an antiviral peptide, wherein in RY1Y2Y2KY1Y3YRY2Y2Y1Y1Y2LKRLY2, Y1 is R, K, D or E, Y2 is I, L, F or W, and Y3 is I or V.

In one embodiment, the antiviral peptide may consist of the amino acid sequence of RKFFKKIYRFFRKLLKRL (SEQ ID NO: 2).

In one embodiment, the antiviral peptide may consist of the amino acid sequence of RLAAKCAARFAEHGLKRG (SEQ ID NO: 3).

In one embodiment, the present invention relates to a composition for preventing or treating a viral disease comprising an antiviral peptide as an active ingredient.

In one embodiment, each of the antiviral peptides of the present invention or a combination thereof can prevent or treat an antiviral disease.

In one embodiment, the antiviral disease of the present invention may be caused by "a virus having a lipid bilayer envelope (or membrane)", wherein the virus is Bunyaviridae, Coronaviridae , Filoviridae, Flaviviridae, Hepadnaviridae, Herpesviridae, Orthomyxoviridae, Poxviridae, Rhabdoviridae (Rhabdoviridae), retroviridae (Retroviridae), togaviridae (Togaviridae), or may be a virus belonging to a family such as Herpesviridae (Herpesviridae), as another example, verniaviridae (Bunyaviridae) ) belonging to the family Sin Nombre Hantavirus and the like; Coronaviruses involved in various acute respiratory syndromes belonging to the Coronaviridae family, and the like; Ebola virus, Marburg virus, etc. belonging to the family Filoviridae; West Nile virus, Yellow Fever virus, Dengue Fever virus, Hepatitis C virus, etc., belonging to the Flaviviridae family; Hepatitis B virus belonging to the family Hepadnaviridae and the like; Herpes Simplex 1 virus, Herpes Simplex 2 virus, etc. belonging to the Herpesviridae family; Influenza virus belonging to the family Orthomyxoviridae and the like; Smallpox virus, Vaccinia virus, Molluscumcontagiosumvirus, Monkeypox virus, etc. belonging to the family Poxviridae; Rabies virus belonging to the family Rhabdoviridae and the like; HIV (Human Immunodeficiency virus), etc. belonging to the retroviridae family; Chikungunya virus belonging to the family Togaviridae and the like; It may be any one or more selected from the group consisting of Pseudorabies virus, HHV virus, and influenza virus belonging to the family Herpesviridae.

In the present invention, the term "prevention" refers to any action that suppresses or delays the spread and recurrence of a disease caused by a virus by administration of the pharmaceutical composition according to the present invention, and "treatment" means the antiviral peptide of the present invention , or a pharmaceutically acceptable salt thereof, or any action of improving or beneficially changing the symptoms of a viral disease by administration of a composition comprising the same. Those of ordinary skill in the art to which the present invention pertains, with reference to the data presented by the Korean Medical Association, etc., know the exact standard of the disease for which the composition of the present application is effective, and can determine the degree of improvement, improvement and treatment will be.

The term "therapeutically effective amount" used in combination with an active ingredient in the present invention means an amount effective to prevent or treat a target disease, and the therapeutically effective amount of the composition of the present invention depends on several factors, for example, the administration method. , the target site, the patient's condition, etc. Therefore, when used in the human body, the dosage should be determined as an appropriate amount in consideration of both safety and efficiency. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. These considerations in determining effective amounts are found, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; and E.W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level is determined by the patient's Health status, viral disease, cause of viral disease outbreak, severity, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment period, factors including drugs used in combination or concurrently; It may be determined according to factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention may include a carrier, diluent, excipient, or a combination of two or more commonly used in biological agents. As used herein, the term “pharmaceutically acceptable” refers to exhibiting non-toxic properties to cells or humans exposed to the composition. The carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition, for example, Merck Index, 13th ed., Merck & Co. Inc. Compounds described in , saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components can be mixed and used, and if necessary, other antioxidants, buffers, bacteriostats, etc. Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable dosage form such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets. Furthermore, it can be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).

In one embodiment, the pharmaceutical composition may be one or more formulations selected from the group including oral formulations, external preparations, suppositories, sterile injection solutions and sprays, and oral or injection formulations are more preferred.

As used herein, the term "administration" means providing a predetermined substance to a subject or patient by any suitable method, and parenteral administration (eg, intravenous, subcutaneous, intraperitoneal) according to a desired method Alternatively, it can be applied locally as an injection formulation) or orally administered, and the dosage varies according to the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease. Liquid formulations for oral administration of the composition of the present invention include suspensions, internal solutions, emulsions, syrups, etc., and various excipients, such as wetting agents, sweetening agents, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. and the like may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. The pharmaceutical composition of the present invention may be administered by any device capable of transporting an active substance to a target cell. Preferred administration methods and formulations are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, drip injections, and the like. For injection, aqueous solvents such as physiological saline and Ringel's solution, vegetable oil, higher fatty acid esters (eg, ethyl oleate), and non-aqueous solvents such as alcohols (eg, ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) Stabilizers for preventing deterioration (e.g., ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH control, to inhibit the growth of microorganisms Pharmaceutical carriers such as preservatives (eg, phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) may be included.

As used herein, the term "individual" means monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, It refers to all animals, including rabbits or guinea pigs, and by administering the pharmaceutical composition of the present invention to an individual, the above diseases can be effectively prevented or treated. The pharmaceutical composition of the present invention may be administered in parallel with a conventional therapeutic agent.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate. , lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, Calcium stearate, sucrose, dextrose, sorbitol and talc and the like may be used. The pharmaceutically acceptable additive according to the present invention is preferably included in an amount of 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

The present invention will be described in more detail through the following examples. However, the following examples are only intended to embody the contents of the present invention, and the present invention is not limited thereto.

Example 1. Preparation of amphiphilic peptides

The present inventors analyzed the physicochemical properties of M2 protein-derived peptides and existing peptides as shown in Table 1 below in order to select peptides suitable for the purpose of the present invention in order to construct an antiviral peptide using an amphiphilic peptide.

As a result, the amphiphilic peptide (SEQ ID NO: 1) derived from the M2 protein of the influenza virus was selected and named as M2 AH. Among the amino acid sequences of the M2 protein derived from influenza A virus of various species, the sequence corresponding to M2 AH was selected and compared to derive the general formula of the peptide of [Equation 1] below.

[Equation 1]

RLX1X2KX3X4YRX5X6X7X8GLKRG

In Formula 1, X1 is S or F, X2 is S or F, X3 is S or C, X4 is I or V, X5 is F, I, R, L or H, and X6 is F or L, X7 is E or K, and X8 is H or Y.

In addition, combinations of sequences corresponding to the general formula are summarized according to the amino acid sequence in Table 2 below.

In addition, to confirm the hypothesis that an increase in amphiphilicity, that is, an increase in the hydrophobic moment, is effective to improve the selectivity for the outer membrane of the virus, the general formula of the peptide corresponding to the following [Equation 2] was derived to actually select such a peptide.

[Equation 2]

RY ₁ Y ₂ Y ₂ KY ₁ Y ₃ YRY ₂ Y ₂ Y ₁ Y ₁ Y ₂ LKRLY ₂

In Formula 2, Y ₁ is R, K, D or E, Y ₂ is I, L, F or W, and Y ₃ is I or V.

In the helix structure of the existing M2 AH (SEQ ID NO: 1), the hydrophilicity of the amino acid located on the hydrophilic face was increased, and a sequence change was induced to increase the hydrophobicity of the amino acid of the hydrophobic face, thereby maximizing the hydrophobicity moment. wanted to

As a result, a peptide corresponding to SEQ ID NO: 2 among the sequences derivable from the general formula of [Formula 2] was synthesized and used in the experiment of the present invention, and was named M2 MH (SEQ ID NO: 2). In addition, peptides that can be derived from Formula 2 in addition to M2 MH (SEQ ID NO: 2) are also included in the scope of the present invention.

Finally, as a control for the results of M2 AH (SEQ ID NO: 1) and M2 MH (SEQ ID NO: 2), the peptide represented by SEQ ID NO: 3 was synthesized and used, which was named M2 NH (SEQ ID NO: 3).

The amino acid sequence and helix structure schematic diagram of the three types of peptides derived from M2 protein used in the present invention, M2 AH (SEQ ID NO: 1), M2 MH (SEQ ID NO: 2), and M2 NH (SEQ ID NO: 3) are as shown in FIG. and, each amino acid in FIG. 1 was expressed differently according to characteristics. (Yellow: non-polar, blue: positive charge, red: negative charge, gray: neutral, and the direction and length of the arrow indicate the direction and magnitude of the hydrophobic moment)

Example 2. Analysis of antiviral activity of antiviral peptides

2-1. Plaque reduction assay

In order to quantitatively measure the antiviral effect of M2 protein-derived peptides, plaque measurement was performed.

To this end, MDCK was cultured to 100% confluency in a 6-well plate and then infected with 100 PFU of A/Puerto Rico/8/34 virus at room temperature for 1 hour.

In the case of the peptide-treated experimental group, the virus was treated with a peptide at a predetermined concentration for 1 hour before cell infection, and cultured at 37° C., 5% CO ₂ . After infecting the cells with the virus, wash thoroughly with PBS, and apply an overlay medium composed of 1% (w/v) agarose, 1X Dulbecco's modified Eagle's medium (Hyclone), and 2μg/mL tosyl phenylalanyl chloromethyl ketone (TPCK)-treated trypsin. Treated and incubated at 37 ℃, 5% CO ₂ for 3 days. After incubation, cells were fixed with 4% formaldehyde and stained with 0.5% crystal violet for observation and analysis.

As a result, as shown in Figure 2, the control group M2 NH (SEQ ID NO: 3) did not induce plaque reduction at any concentration, whereas M2 AH (SEQ ID NO: 1) and M2 MH (SEQ ID NO: 2) were respectively 870 nM and It was confirmed that it showed an IC ₅₀ of 53 nM (concentration induced 50% reduction) to significantly reduce plaque. In particular, M2 MH (SEQ ID NO: 2) showed a value 16.4 times lower than that of M2 AH (SEQ ID NO: 1), confirming that it had excellent antiviral efficacy.

2-2. Broad Effects of M2 Protein-derived Peptides

A/Sydney/5/97 (H3N2), A/X31 (H3N2), A/aquatic bird/Korea/w81/2005 (H5N2), and A /Swine/Iowal/15/30 (H1N1) A plaque reduction assay was performed in the same manner as in Example 2-1 for a total of four influenza viruses.

As a result, as shown in FIG. 3A, the IC ₅₀ for each virus was 58 nM, 16 nM, 66 nM, and 53 nM in that order, and M2 MH (SEQ ID NO: 2) was found in each virus regardless of the virus species. It was confirmed that there is an antiviral effect of a similar aspect.

2-3. Cytotoxicity Assessment (MTT ASSAY)

To analyze the cytotoxicity of the peptides, a cell proliferation response assay (MTT assay) was performed.

First, MDCK was cultured in a 96-well plate at 37° C. to 100% confluency, followed by FBS and 100 U/mL penicillin G, 100 μg/mL streptomycin and 0.25 μg/mL amphotericin B). Treated with 3 synthetic peptides diluted with MEM containing the and cultured for 48 hours.

Thereafter, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used for staining for 2 hours, and dimethylsulfoxide (DMSO) was treated for 2 hours to measure absorbance at 570 nm.

As a result, as shown in FIG. 3B , M2 AH (SEQ ID NO: 1) and M2 MH (SEQ ID NO: 2) showed CC ₅₀ (concentrations that induced 50% cytotoxicity) of 70.3 mM and 19.2 mM, respectively.

2-4. Modification of lipid bilayer by peptide derived from M2 protein

In order to observe the modification of the lipid bilayer by the peptides, liposomes using phospholipids were prepared and the modification was observed.

First, for liposome preparation, a chloroform stock solution containing 25 mM 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC, Avanti Polar Lipids, Alabaster, AL) was treated with nitrogen gas to prepare a lipid film. After vacuuming overnight, the lipid film was dissolved in PBS and vortexed for at least 5 minutes to obtain an aqueous lipid solution.

The freeze-thaw cycle was repeated 5 times with alternating liquid nitrogen and 42 °C water samples for the obtained lipid solution. After that, an extrusion step of passing a polycarbonate membrane (100 nm pores, Avanti Polar Lipids, Alabaster, AL) 21 times was performed using two gas-tight syringes (250 μL, Avanti Polar Lipids, Alabaster, AL). The extruded liposome sample was stored at 4° C. and then used for the experiment.

Liposomes with a diameter of 100 nm were prepared by the same process as above, mixed with peptides of each concentration, and reacted at room temperature for 5 minutes. To measure the diameter change of each sample, DynaPro NanoStar DLS (Wyatt Technologies, Goleta, CA, USA) equipment was used, and each sample was measured and analyzed 10 times at room temperature.

As a result, as shown in Figure 4A, M2 NH control (SEQ ID NO: 3) did not induce any change in liposome diameter, whereas M2 AH (SEQ ID NO: 1) and M2 MH (SEQ ID NO: 2) increased the concentration-dependent diameter. In particular, when M2 MH (SEQ ID NO: 2) at a concentration of 50 mM or more was treated, severe deformation was induced, and no significant diameter was measured.

In addition, as shown in Fig. 4B, as a result of analyzing quantitative changes based on the diameter of the liposome before peptide treatment, M2 AH (SEQ ID NO: 1) and M2 MH (SEQ ID NO: 2) induce an increase in diameter close to 2 times. It was confirmed that it had a direct effect on the lipid bilayer.

2-5. Viral envelope damage and modification by peptides derived from M2 protein

Transmission electron microscope analysis was performed to confirm the action of the peptide on the lipid bilayer envelope of the actual influenza virus.

First, A/Puerto Rico/8/34 virus was treated with a peptide of a predetermined concentration at room temperature for 1 hour. Each sample was treated on a carbon-coated nickel grid (TED PELLA Inc., Redding, CA, USA) for 3 minutes, washed twice with sterile distilled water, and stained with uranyl acetate (UA) for 1 minute, followed by TEM analysis.

As a result, as shown in FIG. 5A, a round shape with a diameter of about 100 nm was observed when no treatment was performed on the virus. On the other hand, in the case of the M2 AH (SEQ ID NO: 1) or M2 MH (SEQ ID NO: 2) treatment group, a phenomenon in which stems extend out of a round shape was observed.

In addition, as shown in Fig. 5B, it was confirmed that the virus appearance change became more severe as the concentration of each peptide increased.

As shown in FIG. 5C , when the concentration of the peptide was increased, a change from several short stems to one long stem was observed.

Through the above results, the antiviral effect of the M2 protein-derived peptide of the present invention was confirmed by confirming that the peptides damage and modify not only the artificially produced lipid bilayer but also the coat of an actual infectious virus.

<110> Research & Business Foundation SUNGKYUNKWAN UNIVERSITY <120> Antiviral peptide from M2 protein and the use thereof <130> PN1907-356 <160> 27 <170> KoPatentIn 3.0 <210> 1 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide M2 AH <400> 1 Arg Leu Phe Phe Lys Cys Ile Tyr Arg Phe Phe Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 2 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide M2 MH <400> 2 Arg Lys Phe Phe Lys Lys Ile Tyr Arg Phe Phe Arg Lys Leu Leu Lys 1 5 10 15 Arg Leu <210> 3 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide M2 NH <400> 3 Arg Leu Ala Ala Lys Cys Ala Ala Arg Phe Ala Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 4 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 4 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Phe Phe Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 5 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 5 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Ile Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 6 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 6 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Arg Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 7 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 7 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Leu Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 8 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 8 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Arg Phe Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 9 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 9 Arg Leu Ser Ser Lys Ser Ile Tyr Arg His Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 10 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 10 Arg Leu Ser Ser Lys Cys Val Tyr Arg Phe Phe Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 11 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 11 Arg Leu Ser Ser Lys Cys Val Tyr Arg Ile Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 12 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 12 Arg Leu Ser Ser Lys Cys Val Tyr Arg Arg Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 13 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 13 Arg Leu Ser Ser Lys Cys Val Tyr Arg Leu Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 14 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 14 Arg Leu Ser Ser Lys Cys Val Tyr Arg Arg Phe Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 15 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 15 Arg Leu Ser Ser Lys Cys Val Tyr Arg His Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 16 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 16 Arg Leu Phe Phe Lys Ser Ile Tyr Arg Phe Phe Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 17 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 17 Arg Leu Phe Phe Lys Ser Ile Tyr Arg Ile Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 18 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 18 Arg Leu Phe Phe Lys Ser Ile Tyr Arg Arg Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 19 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 19 Arg Leu Phe Phe Lys Ser Ile Tyr Arg Leu Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 20 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 20 Arg Leu Phe Phe Lys Ser Ile Tyr Arg Arg Phe Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 21 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 21 Arg Leu Phe Phe Lys Ser Ile Tyr Arg His Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 22 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 22 Arg Leu Phe Phe Lys Cys Val Tyr Arg Phe Phe Glu His Gly Leu Lys 1 5 10 15 Arg Gly <210> 23 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 23 Arg Leu Phe Phe Lys Cys Val Tyr Arg Ile Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 24 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 24 Arg Leu Phe Phe Lys Cys Val Tyr Arg Arg Leu Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 25 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 25 Arg Leu Phe Phe Lys Cys Val Tyr Arg Leu Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly <210> 26 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 26 Arg Leu Phe Phe Lys Cys Val Tyr Arg Arg Phe Lys Tyr Gly Leu Lys 1 5 10 15 Arg Gly <210> 27 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Antiviral peptide <400> 27 Arg Leu Ser Ser Lys Ser Ile Tyr Arg Ile Phe Lys His Gly Leu Lys 1 5 10 15 Arg Gly

Claims (13)

delete delete delete delete delete delete delete delete delete An antiviral peptide comprising the amino acid sequence of RKFFKKIYRFFRKLLKRL represented by SEQ ID NO: 2.
The antiviral peptide of claim 10, wherein the antiviral peptide inhibits infection of influenza A virus.
A pharmaceutical composition for the prevention or treatment of respiratory diseases caused by influenza A virus, comprising the antiviral peptide of claim 10 as an active ingredient. delete

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