KR101675462B1 - Pharmaceutical composition for antivirus comprising base, nucleoside or nucleotide, as active component - Google Patents

Abstract

The present invention relates to a base, nucleoside, or nucleotide having an effective antiviral inhibitory activity against viruses such as rhinovirus, enterovirus or influenza virus having few side effects and having many subtypes.

Description

[0001] The present invention relates to an antiviral pharmaceutical composition comprising a nucleotide, a nucleotide or a nucleotide as an active ingredient,

The present invention relates to bases, nucleosides or nucleotides useful as antiviral agents.

Viruses belonging to the genus Picornaviridae have a + strand RNA as its genome, RNA genome is only 7500nt, and the virus has a diameter of only 20-30nm. Small viruses include rhinovirus, enterovirus, cardiovirus, aphthovirus, hepatovirus, and the like.

Medicines inhibiting rhinovirus include Tremacamra (Truner RB et al. 1999, JAMA 281: 1797-1804), a drug that inhibits the binding of rhinovirus to ICAM-1, which is a sICAM-1 type, Agents having the function of inhibiting 3C protease of virus (HRV) [Wang, QM, 2001. Prog. drug Res. 229-253; Chen DH. Et al. 2003. Bioorg Med Chem Lett 13: 3531-3536 .; Dragovich PS et al. 2002. J Med Chem 45: 1607-1623), agents that react with the capsid of virus include WIN 51711 (Otto MJ et al. 1985. Antimicrob Agents Chemother 27 (6): 883-886), Pirodavir (Brown RN et al., 2005. Bioorg Med Chem. Lett. 15 (8): 2051-2055], Pleconaril [Pevear DC et al., 1999. Antimicrob Agents Chemother 43 (9): 2109-2115] Enviroxime [Carraso L. 1994. 64: 215-290] are being developed.

However, Tramakam is a spray-type formulation that requires frequent daily administration and has a low efficacy. HRV 3C protease inhibitors have been shown to be efficacious against HRV 14 in the in vitro assay [Chen DH. Et al., 2003. Bioorg Med Chem Lett 13: 3531-3536], but did not yield significant results in monkey infestation models [ Dragovich PS et al. 2002. J Med Chem 45: 1607-1623]. Although pyrodavir inhibits the growth of several serotypes of rhinovirus, it has low pharmacodynamic value and is easily hydrolyzed. Although plcnaryl has a wide range of applications including rhinovirus and enterovirus [Hsiung GD & Wang JR 2000. J Microbiol Immunol Infect 33 (1): 1-8], drug interactions, marginal efficacy, It is not approved by the US FDA. RNAi has also been recently studied as a treatment for rhinovirus [Phipps et al. 2004. Antiviral Res. 61: 49-55], there is a disadvantage in that target RNAi must be prepared differently for more than 100 kinds of rhinovirus types. Ribavirin is a type of compound that binds to triosolacarboxamide in the ribose sugar, and FMD virus [Arias A et al. 2008, J Virol 82 (4): 12346-12355), which causes foot-and-mouth disease while belonging to picornaviride, (Fengqin L et al. 2010 J Ethnopharmacol 127 (2): 221-228), a report of antiviral activity in rhinovirus [Choi HJ et al 2010 J med Food 13 (2): 326-328) (Jain MK & Zoellner C 2010 Expert Opin Pharmacother 11 (4): 673-683), which is used as a therapeutic agent, such as interferon, for infectious diseases of hepatitis C virus, However, ribavirin has been known to aggravate heart disease by causing serious anemia, and it also has drawbacks that cause side effects such as tiredness and headache [MacNicholas R & Norris S 2010 Aliment Pharmacol Ther 31 (9): 929-937] .

In addition, the antiviral activity of a modified derivative in which the structure of a nucleotide or a nucleoside is modified (Japanese Patent Publication No. 3032576, Japanese Patent Application Publication No. 2006-521320, Korean Patent Publication No. 2006-0127906, Korean Patent Publication No. 2004-0054775, De Clercq E et al., 1986. Antimicro. Ahebts & Chem. 29 (3): 482-487), but the antiviral activity of unmodified nucleotides or nucleosides has not been studied in the literature.

On the other hand, the influenza virus (Influenza virus) belongs to the family Orthomyxoviridae and is accompanied by influenza. Currently, Tamiflu and Relenza are recognized as medicines for anti-influenza virus. Tamiflu is a pill or liquid, and the respiratory tract is inhaled. Relenza is inhaled, which is very uncomfortable for patients with respiratory disease and has the disadvantage of causing severe side effects such as Stevens-Jones disease or toxic skin necrosis in people taking Tamiflu.

Under these circumstances, the inventors of the present invention have made efforts to find a compound having antiviral activity against viruses showing many subspecies such as rhinovirus, enterovirus, and influenza virus and having few side effects. As a result, Nucleosides and nucleotides such as adenine diphosphate, adenosine diphosphate, adenosine triphosphate and the like have selective antiviral activity against rhinovirus, enterovirus and influenza virus, and have completed the present invention.

The present invention is to provide a nucleotide, nucleoside, or nucleotide having an effective antiviral inhibitory activity against a virus such as rhinovirus, enterovirus or influenza virus having few side effects and having many subtypes.

In one embodiment, the present invention provides an antiviral agent comprising at least one selected from the group consisting of adenine, guanine, uracil, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate and adenosine triphosphate as an active ingredient ≪ / RTI >

As used herein, "adenine" and "guanine" are purine bases and refer to one of the basic materials constituting DNA and RNA, and "uracil" and "cytosine" It means one of the substances.

As used herein, "adenosine" is a structure in which a ribose sugar is bound to adenine as a purine nucleoside, and plays an important role in energy transfer in vivo in the form of adenosine diphosphate and adenosine triphosphate.

As used herein, "guanosine" is a purine nucleoside having a structure in which a ribose sugar is bound to guanine. Guanosine is readily phosphorylated and plays an important role in the synthesis and energy transfer of nucleic acids in vivo in the form of cyclic guanosine phosphoric acid, guanosine phosphorus acid and guanosine triphosphate.

The term " uridine "used in the present invention is a structure in which a ribose sugar is bound to uracil, and is used as a synthesis substrate for RNA." Cytidine " .

The bases of said adenine, guanine, uracil and cytosine according to the present invention; nucleosides of adenosine, guanosine, uridine and cytidine; And nucleotides of adenosine diphosphate and adenosine diphosphate can be purchased commercially, directly synthesized, or extracted from natural products, etc., but are not limited thereto.

The bases, nucleosides and nucleotides of the present invention may be used alone or in combination or combination with other pharmaceutically active compounds.

The term "antiviral" used in the present invention means an action that specifically inhibits the proliferation of virus by specifically inhibiting the cytopathic effect of the virus. Preferably, the virus may be a rhinovirus or an enterovirus. More preferably, the rhinovirus may be any one of rhinovirus type 2, type 3, type 6, type 14, type 15, and type 40, and the enterovirus may be type 71 enterovirus have.

As used herein, the term " rhinovirus "means a virus belonging to the genus Picornavirida, which refers to a small virus having a virus diameter of only 20-30 nm. Rhinoviruses are divided into rhinovirus type A and rhinovirus type B viruses, all of which mainly cause respiratory tract infections, causing more than 50% of all colds. The main symptoms of infection are sinusitis, runny nose, otitis media, and chronic bronchitis.

As used herein, "enterovirus" is a virus belonging to the genus Picornavirida together with rhinovirus, causing diseases such as myocarditis, encephalitis, and enterovirus 71.

Since the bases, nucleosides and nucleotides according to the present invention exhibit selective proliferation inhibitory activity against viruses, they can be usefully used as pharmaceutical compositions for antiviral use.

In particular, the bases, nucleosides and nucleotides of the present invention have antiviral inhibitory activity against various subtypes of rhinovirus, so that target RNAi can be produced for each of more than 100 types of rhinoviruses, It is possible to eliminate disadvantages such as the viral activity and the necessity of using various complex preparations.

In addition, there are disadvantages such as serious anemia of ribavirin used as an antiviral agent, side effects of headache, side effects of headache, adverse effects of Stevens-Johnson disease or toxic skin necrosis of Tamiflu used as an anti-influenza virus agent, And has an activity of selectively inhibiting virus proliferation.

In a specific embodiment, adenine, guanine, and uracil inhibited virus proliferation at a concentration of 100 ug / ml to a degree similar to ribavirin, a common antiviral agent (about 100%) for rhinovirus type 2 (FIG. 1).

In another specific embodiment, for rhinovirus type 6, guanosine and cytidine showed significantly higher antiviral activity than ribavirin (TCID ₅₀ 25.58 ug / ml) as a control at 0.87 ug / ml of TCID ₅₀ and 0.68 ug / ml, respectively (Table 2 and Fig. 4).

In another specific embodiment, adenosine, adenosine diphosphate and adenosine triphosphate inhibit the proliferation of rhinoviruses 2, 3, 6, 14, 15 and 40 at concentrations of 100 ug / (Table 3 and Figs. 6 to 11). Also, in another specific embodiment, a significant antiviral action of adenine to enterovirus type 71 was identified (Table 4 and Figure 12).

In another aspect, the present invention relates to a pharmaceutical composition for anti-influenza virus comprising at least one of adenine and adenosine as an active ingredient.

Preferably, the influenza virus may be influenza A virus type.

In a specific embodiment, adenine and adenosine were found to inhibit virus proliferation to a similar degree (about 100%) to that of Tamiflu used as a general anti-influenza virus agent at a concentration of 100 ug / ml (see Tables 5,6 and 14 , 15).

In another aspect, the present invention relates to a method for preventing or treating a viral disease comprising administering a composition of the present invention to a subject having a possible or suspected occurrence of a viral infectious disease in a pharmaceutically effective amount.

The term " prophylactic " as used herein refers to any act that inhibits or delays a viral infectious disease by administration of a composition comprising a base, nucleoside or nucleotide of the present invention. The term " treatment " as used in the present invention means all actions that improve or ameliorate symptoms of a disease by administration of a composition comprising the base, nucleoside or nucleotide.

As used herein, the term " individual " means all animals including humans that have already developed or may have developed a viral infection disease. By administering the composition of the present invention to an individual, the disease can be effectively prevented and treated.

The composition of the present invention is administered in a pharmaceutically effective amount. The term " pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment for medical treatment and the effective dosage level will vary depending on the species and severity, Sex, type of viral infectious disease, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, excipient or diluent in addition to the above-described effective ingredient. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical compositions of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories or sterilized injection solutions according to a conventional method have. In detail, when formulating, it can be prepared by using diluents or excipients such as fillers, weighing agents, binders, humectants, disintegrants, surfactants and the like which are generally used. Solid formulations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration, liquid paraffin, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method, and the dose may vary depending on the condition and the weight of the patient, Dose, preferably 20 to 100 mg / kg, per day, and is administered at a predetermined time interval according to the judgment of a doctor or pharmacist, It may be administered several times a day, preferably once or four times, and may be suitably selected by those skilled in the art.

In another aspect, the present invention provides an antiviral agent comprising at least one selected from the group consisting of adenine, guanine, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate and adenosine triphosphate as an active ingredient ≪ / RTI >

Adenine, guanine, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate, adenosine triphosphate or mixtures thereof may be added to the food composition for the purpose of preventing or ameliorating viral infection. When such a component is used as a food additive, it may be added as it is or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention. In addition, the composition of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. The proportion of such flesh is not critical but is generally selected in the range of 0.01 to 10 parts by weight per 100 parts by weight of the composition of the present invention. These components can be used independently or in combination.

In another aspect, the present invention provides a method for screening for viruses comprising at least one selected from the group consisting of adenine, guanine, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate and adenosine triphosphate as active ingredients Quasi-drug compositions.

That is, the composition of the present invention may be added to a quasi-drug composition for the purpose of virulence. When adenine, guanine, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate, adenosine diphosphate, or a mixture thereof is used as a quasi-drug additive, it can be added as it is or used in combination with other quasi-drugs or quasi-drug components, And can be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).

Preferably, the quasi-drug composition may be used in the manufacture of a disinfectant cleaner, shower foam, gagrin, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.

In another aspect, the present invention provides an antiviral agent comprising at least one selected from the group consisting of adenine, guanine, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate and adenosine triphosphate as an active ingredient Or feed additive for viral or live viruses.

Since the feed additive has antiviral and antiviral efficacy, the feed additive can be prevented from viral diseases by continuously ingesting the feed additives into poultry, domestic animals and the like, and viral diseases that have already occurred can be improved. Feeds can be classified into various categories according to their nutritional value, main ingredient, distribution, moisture content, processing type and the like, and the feeds can be used for forage, concentrated feed, supplementary feed, protein feed, starch feed, fat feed or fiber feed But is not limited thereto.

The feed additive may additionally contain a carrier such as poultry and livestock. In the present invention, the feed additive may be added as it is or a known carrier, stabilizer and the like may be added. Various nutrients such as vitamins, amino acids and minerals, antioxidants, antibiotics, antibacterial agents and other additives may be added And the shape thereof may be a suitable state such as powder, granule, pellet, suspension and the like. When the feed additive of the present invention is supplied, it can be supplied to poultry, livestock and the like singly or mixed with feed.

In another aspect, the present invention relates to a feed containing the feed additive. The feed additives described above can be used as feeds for various animal feeds, as raw materials and additives for negative water, and can be used as a feed composition containing the feed additives.

Since the base, nucleoside or nucleotide according to the present invention has an effective antiviral inhibitory activity against viruses such as rhinovirus, enterovirus or influenza virus having few side effects and having many subtypes, Can be used as a composition.

Figure 1 is a graph of antiviral activity against rhinovirus type 2 of adenine, guanine, uracil and cytosine, according to one embodiment of the present invention.
Figure 2 is a graph of antiviral activity against rhinovirus type 6 of adenine, guanine, uracil and cytosine, according to one embodiment of the present invention.
Figure 3 is a graph of antiviral activity against rhinovirus type 2 of adenosine, guanosine, uridine and cytosine, according to one embodiment of the present invention.
Figure 4 is a graph of antiviral activity against rhinovirus type 6 of adenosine, guanosine, uridine and cytosine, according to one embodiment of the present invention.
Figure 5 is a graph of antiviral activity against rhinovirus type 14 of adenosine, guanosine, uridine and cytosine, according to one embodiment of the present invention.
Figure 6 is a graph of antiviral activity against rhinovirus type 2 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
Figure 7 is a graph of antiviral activity against rhinovirus type 3 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
Figure 8 is a graph of antiviral activity against rhinovirus type 6 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
Figure 9 is a graph of antiviral activity against rhinovirus type 14 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
Figure 10 is a graph of antiviral activity against rhinovirus type 15 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
Figure 11 is a graph of antiviral activity against rhinovirus type 40 of adenosine, adenosine diphosphate and adenosine triphosphate, according to one embodiment of the present invention.
12 is a graph showing antiviral activity against enterovirus type 71 of adenine, guanine, uracil and cytosine according to an embodiment of the present invention.
FIG. 13 is a graph showing antiviral activity against enterovirus type 71 of adenosine triphosphate according to an embodiment of the present invention.
Figure 14 is a graph of antiviral activity against influenza virus A forms of adenine, guanine, uracil and cytosine, according to one embodiment of the present invention.
Figure 15 is a graph of antiviral activity against influenza virus A forms of adenosine, guanosine, uridine, and cytidine, according to one embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited by these examples.

<Materials>

The test compounds adenine, guanine, uracil, cytosine, adenosine, guanosine, uridine, cytidine, adenosine diphosphate (ADP) and adenosine triphosphate were purchased from Sigma Respectively.

As a control, ribavirin was purchased from Sigma and diluted in cell culture solution.

Viruses used for measuring the virus-inhibiting activity of bases, nucleosides and nucleotides according to the present invention are rhinovirus type 2 (HRV-2), rhinovirus type 3 (HRV-3), rhinovirus type 6 (HRV- 6), rhinovirus 14 (HRV-14), rhinovirus 15 (HRV-15), rhinovirus 40 (HRV-40), enterovirus 71 (EV71) and influenza A virus / 34) was used. Each virus was purchased from ATCC, and the viral activity was measured after incubation and stored at -70 ° C.

Example  1: adenine, guanine, Uracil  And Cytosine  Antiviral activity test for rhinovirus

Vero cells (2 x 10 ⁴ ) were placed in each well of 96-well plates and cultured for 24 hours. After 24 hours, the culture supernatant of each well was removed, and a rhinovirus type 2 virus solution titrated with TCID ₅₀ was added to each well. The concentrations of adenine, guanine, uracil and cytosine were 0.1 μg / ml, ug / ml and 100 ug / ml. Adenine, guanine, uracil and cytosine were assayed for their ability to inhibit the proliferation of viruses according to the method disclosed in Korean Patent No. 10-0682069.

Specifically, 100 μl of a 70% acetone solution was added to each well after the viral infection test was completed, and the mixture was allowed to stand at 4 ° C for 1 hour and washed several times with distilled water. After drying at room temperature, 100 μl of 0.4% (w / v) sulforhodamine B solution in 1% (v / v) acetic acid was added and stained for 30 minutes. The SRB staining solution not bound to the cells was washed several times with 1% (v / v) acetic acid and dried again. 100 μl of a 10 mM Tris solution (pH 10.5) was added to each well to sufficiently dissolve the stain associated with the cells, and the absorbance was measured at 560 nm. Each treatment group was divided into two groups: (A) adenine, guanine, cytosine, uracil only (B), virus only (C), virus and nucleotide (D) , And the cell viability (%) of adenine, guanine, uracil and cytosine was calculated as shown in the following Equation 1, and the inhibitory activity (%) of adenine, guanine, uracil and cytosine for virus growth was calculated by the following formula. The results are shown in Table 1 and Figs. 1 and 2. The statistics of the results are calculated by the mean value and the standard deviation of the results of three experiments performed under the same conditions.

[Equation 1]

Cell viability (%) = A / B X 100

&Quot; (2) &quot;

Virus Proliferation Inhibition (%) = (D-C) / (B-C) X 100

division
TCID ₅₀ (ug / ml) Adenine Guanine Uracil Cytosine Rhinovirus type 2 1.43 3.92 14.6 8.83 Rhinovirus type 6 39.4 4.1 33.8 114.6

As a result, as shown in Table 1 and FIG. 1, adenine, guanine, uracil and cytosine generally increased anti-rhinovirus activity with increasing concentrations, and the concentration of 100 ug / ml of the anti- To inhibit the proliferation of the rhinovirus type 2 at a similar level to that of the rhinovirus type 2 strain.

In addition, as shown in Table 1 and Fig. 2, adenine and guanine were confirmed to have anti-rhinovirus activity by inhibiting rhinovirus type 6 at a concentration similar to that of ribavirin used as a control group at a concentration of 100 ug / ml.

Example  2: adenosine, Guanosine , Our Dean  And Cytidine  Antiviral activity test for rhinovirus

6, and 14 were prepared in the same manner as in Example 1, except that adenosine, guanosine, uridine, and cytidine were used instead of adenine, guanine, uracil, Virus activity tests were performed and the results are shown in Table 2 and Figures 3-5.

division
TCID ₅₀ (ug / ml) Adenosine Guanosine Our Dean Cytidine Ribavirin Rhinovirus type 2 27.5 - * 0.88 - * 3.4 Rhinovirus type 6 10.7 0.68 59.4 0.68 25.58 Rhinovirus type 14 79.01 - * - * - * 6.15

* Virus growth inhibition rate is less than 50%.

As a result, as shown in Table 2 and FIG. 3, Uridine showed an inhibitory activity of TCID _{50 of} 0.88 ug / ml against rhinovirus type 2, confirming that rhinovirus type 2 inhibitory activity is superior to ribavirin used as a control.

In addition, as shown in Table 2 and FIG. 4, both guanosine and cytidine exhibited an inhibitory activity of 0.68 ug / ml of TCID ₅₀ with respect to the rhinovirus 6 type, and the inhibitory activity against rhinovirus 6 type .

In particular, it was confirmed that adenosine inhibits the proliferation of rhinovirus at a similar level (100%) to that of the antiviral drug ribavirin used as the control group at a concentration of 100 ug / ml for rhinovirus type 2 and type 6.

As shown in Table 2 and FIG. 5, adenosine inhibited the proliferation of rhinovirus type 14 at a concentration of 79.01 ug / ml of TCID ₅₀ for rhinovirus type 14.

Example  3: adenosine, adenosine Phosphoric acid  And adenosine Triple  Antiviral activity test for rhinovirus

Type 6, type 14, type 15, adenine diphosphate, adenosine diphosphate, adenosine diphosphate, adenosine diphosphate, adenosine diphosphate and adenosine diphosphate were used in place of adenosine, adenosine diphosphate and adenosine diphosphate instead of adenine, guanine, Antiviral activity tests were carried out for Type-40 and Type-40, and the results are shown in the following Table 3 and Figures 6 to 11.

division
TCID ₅₀ (ug / ml) Adenosine Adenosine diphosphate Adenosine triphosphate Ribavirin Rhinovirus type 2 27.5 19.6 13.3 3.4 Rhinovirus type 3 252.70 55.6 69.5 8.74 Rhinovirus type 6 10.7 72.3 107.7 62.3 Rhinovirus type 14 79.02 71.8 90.5 9.38 Rhinovirus 15 type 35.3 31.3 53.2 23.4 Rhinovirus type 40 63.3 58.1 - * 60.3

* Virus growth inhibition rate is less than 50%.

As a result, as shown in Table 3 and Figs. 6 to 11, adenosine, adenosine diphosphate and adenosine triphosphate were found to be highly resistant to rhinovirus 2, 3, 6, 14 and 15 And 40 &lt; / RTI &gt;

Example  4: adenine, guanine, Uracil  And Cytosine Enterovirus  Antiviral activity test

The antiviral activity test for the enterovirus type 71 of adenine, guanine, uracil and cytosine was carried out using the same method as in Example 1 except that the enterovirus was used instead of the rhinovirus, And Fig. 12.

division
TCID ₅₀ (ug / ml) Adenine Guanine Uracil Cytosine Enterovirus type 71 83.4 - * - * - *

* Virus growth inhibition rate is less than 50%.

As a result, as shown in Table 4 and FIG. 12, in the case of adenine, it was confirmed that anti-enterovirus activity was 83.4 ug / ml of TCID ₅₀ .

Example  5: adenosine Triple Enterovirus  Antiviral activity test

The antiviral activity test for Enterovirus type 71 was performed using the same method as in Example 1, except that adenosine triphosphate and enterovirus were used, and the results are shown in Table 5 and FIG. As a control, water-soluble dexamethasone, beta-cyclodextrin, L-mimosin, isopropyl beta-D-thiogalactoside was used.

As a result, as shown in Fig. 13, effective antiviral activity was observed even at a concentration of 100 ug / ml, which was used as a control group, for water-soluble dexamethasone, beta-cyclodextrin, L-mimosin and isopropyl beta-D-thiogalactoside However, adenosine triphosphate (ATP) significantly inhibited the proliferation of enterovirus type 71.

Example  6: adenine, guanine, Uracil  And Cytosine  Antiviral activity test for influenza virus type A

Anti-influenza virus activity tests of adenine, guanine, uracil and cytosine were performed using the same method as in Example 1 except that influenza virus type A (A / PR / 8/34) was used, Shown in Table 5 and Fig.

division
TCID ₅₀ (ug / ml) Adenine Guanine Uracil Cytosine Tamiflu influenza
Virus type A 47.1 - * - * - * 0.73

* Virus growth inhibition rate is less than 50%.

As a result, as shown in Table 5 and Fig. 14, it was confirmed that the adenine inhibited the proliferation of influenza virus at a concentration of 100 ug / ml similar to that of Tamiflu, an anti-influenza virus therapeutic agent.

Example  7: adenosine, Guanosine , Our Dean  And Cytidine  Antiviral activity test for influenza virus type A

An anti-influenza virus activity test was carried out using the same method as in Example 1 except that influenza virus type A (A / PR / 8/34) and adenosine, guanosine, uridine and cytidine were used, The results are shown in Table 6 and Fig.

division
TCID ₅₀ (ug / ml) Adenosine Guanosine Our Dean Cytidine Tamiflu influenza
Virus type A 9.62 - * - * - * 0.73

* Virus growth inhibition rate is less than 50%.

As a result, as shown in Table 6 and Fig. 15, it was confirmed that adenosine inhibited the proliferation of influenza virus at a concentration of 100 ug / ml to a degree similar to that of Tamiflu, an anti-influenza virus therapeutic agent.

Claims (10)

Wherein the active ingredient is composed of adenosine diphosphate,
A pharmaceutical composition for antiviral therapy for one or more viruses selected from the group consisting of rhinovirus type 6, type 14, and type 40.
Wherein the active ingredient is composed of adenosine triphosphate,
An antiviral pharmaceutical composition for enterovirus type 71.
Wherein the active ingredient is composed of adenosine diphosphate,
A food composition for antiviral therapy for one or more viruses selected from the group consisting of rhinovirus type 6, type 14, and type 40.
Wherein the active ingredient is composed of adenosine triphosphate,
An antiviral food composition for enterovirus type 71.
Wherein the active ingredient is composed of adenosine diphosphate,
The quasi-quadriceps composition for viral infection of one or more viruses selected from the group consisting of rhinovirus type 6, type 14, and type 40.
Wherein the active ingredient is composed of adenosine triphosphate,
Composition for quail virus for enterovirus type 71.
Wherein the active ingredient is composed of adenosine diphosphate,
An antiviral or fowl feed additive for one or more viruses selected from the group consisting of rhinovirus type 6, type 14, and type 40.
Wherein the active ingredient is composed of adenosine triphosphate,
Feed additive for antiviral or fowl virus against enterovirus type 71.
A feed comprising the feed additive according to claim 7 or 8.




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