KR20160005980A - Composition comprising hydrocortisone or pharmaceutically acceptable salts thereof as an active ingredient for anti virus - Google Patents

Abstract

Hydrocortisone of the present invention significantly has antiviral activities with respect to various viruses comprising rhinovirus type 2, rhinovirus type 3, rhinovirus type 5, and hepatitis A virus. Particularly, the hydrocortisone can be usefully used as an active ingredient of a health functional food and a pharmaceutical composition for antiviral activities, by representing significant antiviral activities in comparison with Ribavirin and Acyclovir which are positive control groups.

Description

[0001] The present invention relates to an antiviral composition containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient,

The present invention relates to an antiviral pharmaceutical composition and a health functional food containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

Viruses belonging to the genus Picornaviridae have the RNA of plus strand as its genome, the RNA genome is only 7500 nt, and the virus has a diameter of 20 It is a small virus that is only -30 nm and belongs to rhinovirus, enterovirus, cardiovirus, aphthovirus and hepatovirus.

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) .

The proteases 2A and 3C proteins of the virus belonging to the Picornavirida activate the caspase 3 enzyme and cause apoptosis of the host cell. The offspring viruses generated from host cells that are first infected through the death of host cells are very easily propagated to other cells [Buenz EJ & Lowe CL. Trends in Microbiology. 2006 14 (1): 28-36].

Hydrocortisone, also known as cortisol, has the ability to increase blood sugar and inhibit the immune response. It is currently being used as a remedy for endocrine disorders, rheumatic disorders, and allergic diseases. [RMR Pereira et al. 2001. Bone 28 (5): 484-490] Administration of hydrocortisone lowers caspase 3 activity [Menshanov PN et al., 3, 2013. Physiol Res 62: 205-213].

Accordingly, the present inventors have made efforts to find a compound having antiviral activity, and as a result, have found that hydrocortisone has remarkable antiviral activity against various viruses including rhinovirus type 2, type 3, type 5 and hepatitis A virus Particularly, ribavirin and acyclovir, which are positive control groups, and thus, hydrocortisone can be effectively used as an active ingredient of a pharmaceutical composition for antiviral treatment, thereby completing the present invention.

The present invention provides an antiviral pharmaceutical composition and a health functional food containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

To achieve the above object, the present invention provides an antiviral pharmaceutical composition comprising hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides an antiviral health functional food containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention and treatment of viral diseases containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention is based on the finding that hydrocortisone has remarkable antiviral activity against various viruses including rhinovirus type 2, type 3, type 5 and type A hepatitis viruses and is particularly effective against positive control strains such as ribavirin and ribavirin As compared with acyclovir, it exhibits significant antiviral activity, so that hydrocortisone can be effectively used as an active ingredient of a pharmaceutical composition for antiviral use.

Figure 1 is a graph of antiviral activity against rhinovirus type 2 of hydrocortisone and ribavirin.
Figure 2 is a graph of antiviral activity against rhinovirus type 3 of hydrocortisone and ribavirin.
Figure 3 is a graph of antiviral activity against rhinovirus type 5 of hydrocortisone and ribavirin.
4 is a graph comparing antiviral activities of hydrocortisone and acyclovir against rhinovirus type 2.
FIG. 5 is a graph comparing antiviral activities of hydrocortisone and ribavirin against hepatitis A virus.

As used herein, the term "prophylactic " means any act that inhibits or slows the progression of a viral infectious disease upon administration of the composition of the present invention.

 As used herein, the terms " treatment "and" improvement "refer to all actions by which administration of the composition of the invention improves or alleviates symptoms of a viral infection disease.

The term "administering" as used herein is meant to provide any desired composition of the invention to an individual by any suitable method.

As used herein, the term "individual" means any animal such as a human, a monkey, a dog, a goat, a pig or a mouse having a disease in which symptoms of a viral infection disease can be improved by administration of the composition of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides an antiviral pharmaceutical composition containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

The hydrocortisone is preferably a compound represented by the following formula 1:

[Chemical Formula 1]

.

.

The virus is preferably a virus belonging to the genus Picornaviridae , but is not limited thereto. The virus belonging to the genus Picornavirida may be rhinovirus, enterovirus, cardiovirus Aphthovirus, poliovirus, and hepatovirus, preferably rhinovirus and hepatovirus.

Preferably, the rhinovirus is any one selected from the group consisting of rhinovirus type 2, type 3, and type 5, and the hepatovirus is preferably hepatitis A virus, but is not limited thereto.

In a specific embodiment of the present invention, we compared the antiviral activity of hydrocortisone with that of ribavirin as a positive control to ascertain antiviral activity against rhinovirus, and as a result, In the case of rhinovirus type 2, which is represented by rhinovirus type A, hydrocortisone showed 93.05 ± 1.41% virus inhibition ability at a dose of 100 ㎍ / ml. Thus, ribavirin (68. 67 ± 10.70%) (see Table 1 and Figure 1). In addition, hydrocortisone showed 90.46 ± 0.64% virus inhibition ability at 100 μg / ml at the administration concentration of rhinovirus type 3 represented by rhinovirus type B, and showed superior efficacy to ribavirin (89.05 ± 2.20%), which is a general antiviral agent (See Table 1 and Fig. 2). In addition, for another rhinovirus type 5 represented by rhinovirus type B, hydrocortisone showed 93.94 ± 0.35% virus inhibitory ability at a dose of 100 μg / ml, which is superior to general antiviral drug ribavirin (90.14 ± 0.83%) (See Table 1 and Fig. 3).

In addition, the hydrocortisone of the present invention was confirmed to have antiviral activity against rhinovirus as compared with ascicclovir as a positive control. As a result, hydrocortisone for rhinovirus type 3 was found to be 65.37 ± 3.73 %, While acyclovir (-0.75 + 4.48%) did not show antiviral activity against rhinovirus (see Table 2 and Fig. 4).

In order to confirm the antiviral activity of hydrocortisone of the present invention against hepatitis A virus, ribalin was used as a positive control. As a result, hydrocortisone was administered at a dose of 100 / / ml at 57.55 (Table 3 and Fig. 5). As shown in Table 3 and Fig. 5, the antiviral effect of the present invention was better than that of the common antiviral drug ribavirin (20.44 + 4.64%).

Therefore, the hydrocortisone of the present invention has remarkable antiviral activity against various viruses including rhinovirus type 2, type 3, type 5 and type A hepatitis viruses, and is particularly effective against positive control ribavirin and acyclovir The above-mentioned hydrocortisone can be effectively used as an active ingredient of a pharmaceutical composition for antiviral use.

The present invention encompasses all of the possible solvates, hydrates, which can be prepared therefrom, as well as hydrocortisone or a pharmaceutically acceptable salt thereof.

The compound of the present invention can be used in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the invention can be prepared by a conventional method, for example by dissolving the compound of the invention in an excess of an aqueous acid solution and treating the salt with a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile To precipitate it.

By heating the same amount of hydrocortisone and an acid or alcohol in water, then evaporating the mixture and drying, or by suction filtration of the precipitated salt.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

The therapeutically effective amount of the composition of the present invention may vary depending on a variety of factors, such as the method of administration, the site of administration, the condition of the patient, and the like. Therefore, when used in the human body, the dosage should be determined in consideration of safety and efficacy. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. Such considerations in determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman ' s Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington ' s Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

Compositions of the present invention may also include carriers, diluents, excipients, or a combination of two or more thereof commonly used in biological formulations. The pharmaceutically acceptable carrier is not particularly limited as long as the composition is suitable for in vivo delivery, for example, Merck Index, 13th ed., Merck & Sodium chloride, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components may be mixed and used. If necessary, an antioxidant, a buffer, And other conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into main dosage forms such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules or tablets. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990) in a suitable manner in the art.

The composition of the present invention may further contain one or more active ingredients showing the same or similar functions. The composition of the present invention may contain 0.0001 to 10% by weight, preferably 0.001 to 1% by weight of the above compound, based on the total weight of the composition.

The composition of the present invention may be administered orally or non-orally (for example, intravenously, subcutaneously, intraperitoneally or topically) or orally administered in accordance with a desired method, and the dose may be appropriately determined depending on the patient's body weight, The range varies depending on diet, administration time, method of administration, excretion rate, and severity of the disease. The daily dose of the composition according to the present invention is 0.0001 to 10 mg / ml, preferably 0.0001 to 5 mg / ml, more preferably administered once to several times a day.

In addition, the present invention provides an antiviral health functional food containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

The virus is preferably a virus belonging to the genus Picornaviridae , but is not limited thereto. The virus belonging to the genus Picornavirida may be rhinovirus, enterovirus, cardiovirus Aphthovirus, poliovirus, and hepatovirus, preferably rhinovirus and hepatovirus.

Preferably, the rhinovirus is any one selected from the group consisting of rhinovirus type 2, type 3, and type 5, and the hepatovirus is preferably hepatitis A virus, but is not limited thereto.

The present invention also provides a pharmaceutical composition for the prevention and treatment of viral diseases containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.

The virus is preferably a virus belonging to the genus Picornaviridae , but is not limited thereto. The virus belonging to the genus Picornavirida may be rhinovirus, enterovirus, cardiovirus Aphthovirus, poliovirus, and hepatovirus, preferably rhinovirus and hepatovirus.

Preferably, the rhinovirus is any one selected from the group consisting of rhinovirus type 2, type 3, and type 5, and the hepatovirus is preferably hepatitis A virus, but is not limited thereto.

The viral disease is selected from the group consisting of polio, paralysis, acute hemorrhagic conjunctivitis, viral meningitis, waterborne disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes mellitus, encephalitis, , Chronic obstructive pulmonary disease, pneumonia, sinusitis or otitis media.

Therefore, the hydrocortisone of the present invention has remarkable antiviral activity against various viruses including rhinovirus type 2, type 3, type 5 and type A hepatitis viruses, and is particularly effective against positive control ribavirin and acyclovir The hydrocortisone is useful as an antiviral pharmaceutical composition and an active ingredient of a health functional food.

Hereinafter, the present invention will be described in detail with reference to the following examples.

However, the following examples are illustrative of the present invention, and the scope of the present invention is not limited by the following examples.

< Example  1> Preparation of experimental material

Hydrocortisone, a test compound for measuring the virus-inhibiting activity of the present invention, was purchased from Sigma. Ribavirin and Acyclovir were used as positive control for hydrocortisone, purchased from Sigma, and diluted in cell culture solution.

(HRV-2), rhinovirus type 3 (HRV-3), rhinovirus type 5 (HRV-5), hepatitis A virus Virus was used. Each virus was purchased from ATCC, and the viral activity was measured after incubation and stored at -70 ° C.

< Example  2> Hydrocortisone ( hidrocortisone ) Of rhinovirus ( rhinovirus ) For antiviral activity

<2-1> Ribavirin ( Ribavirin ) And Hydrocortisone Antivirus  Active comparison

In order to confirm the antiviral activity of the hydrocortisone of the present invention against rhinovirus, experiments were carried out as follows. Ribavirin was used as a positive control.

Specifically, HeLa cells (2x10 ⁴ cells) 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 solution of rhinovirus (rhinovirus type 2, rhinovirus type 3 and rhinovirus type 5) titrated with TCID ₅₀ was added to each well. Then, the concentration of hydrocortisone was adjusted to 0.1 μg / ml, 1 占 퐂 / ml, 10 占 퐂 / ml and 100 占 퐂 / ml.

The ability of hydrocortisone to inhibit the proliferation of viruses was measured according to the method disclosed in Korean Patent No. 10-0682069. After the viral infection test was completed, 100 μl of 70% acetone solution was added to each well, followed by standing at 4 ° C for 1 hour and washing 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, the stain bound to the cells was sufficiently dissolved, and the absorbance was measured at 560 nm. Each treatment group was a group (A) treated with nothing, a group treated with hydrocortisone alone (B), a group treated with virus alone (C), and a group treated with virus and hydrocortisone (D) The inhibitory activity (%) was calculated by the following formula (2). The results were 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 rate (%) = (D-C) / (B-C) X 100

As a result, as shown in Table 1 and Figs. 1 to 3, hydrocortisone increased anti-rhinovirus activity as the concentration increased. For rhinovirus type 2, which is represented by rhinovirus type A, And showed 93.05 ± 1.41% virus inhibition ability at ㎍ / ml, showing superior efficacy to ribavirin (68. 67 ± 10.70%) as a general antiviral agent (Table 1 and FIG. 1). In addition, hydrocortisone showed 90.46 ± 0.64% virus inhibition ability at 100 μg / ml at the administration concentration of rhinovirus type 3 represented by rhinovirus type B, and showed superior efficacy to ribavirin (89.05 ± 2.20%), which is a general antiviral agent (Table 1 and Fig. 2). In addition, for another rhinovirus type 5 represented by rhinovirus type B, hydrocortisone showed 93.94 ± 0.35% virus inhibitory ability at a dose of 100 μg / ml, which is superior to general antiviral drug ribavirin (90.14 ± 0.83%) (Table 1 and Fig. 3).

division Antiviral activity (%) at a concentration of 100 / / ml Rhinovirus type 2 Rhinovirus type 3 Rhinovirus type 5 Hydrocortisone 93.05 + 1.41 90.46 ± 0.64 93.94 + 0.35 Ribavirin 68.67 + - 10.70 89.05 ± 2.20 90.14 + 0.83

<2-2> Acyclovir ( Acyclovir ) And Hydrocortisone  Comparison of antiviral activity

In order to confirm the antiviral activity of the hydrocortisone of the present invention against the rhinovirus, antiviral activity tests on the rhinovirus type 2 virus and the rhinovirus type 3 virus were performed using the same method as in Example <2-1> , And ascyclovir was used as a positive control.

As a result, as shown in Table 2 and FIG. 4, hydrocortisone showed 65.37 ± 3.73% at 100 μg / ml for the rhinovirus type 3, while acyclovir (-0.75 ± 4.48% (Table 2 and Fig. 4). &Lt; tb &gt; &lt; TABLE &gt;

division Antiviral activity (%) at a concentration of 100 / / ml Rhinovirus type 2 Rhinovirus type 3 Hydrocortisone 87.4 ± 6.79 65.37 ± 3.73 Acyclovir 36.21 ± 1.62 -0.75 + 4.48

< Example  3> Hydrocortisone ( hidrocortisone ) To confirm antiviral activity against hepatitis A virus

In order to confirm the antiviral activity of hydrocortisone of the present invention against hepatitis A virus, an antiviral activity test was carried out using the same method as in Example <2-1>, and ribalin was used as a positive control.

As a result, as shown in Table 3 and FIG. 5, hydrocortisone showed a virus-suppressing ability of 57.55 + 4.31% at a dose of 100 占 퐂 / ml for hepatitis A virus and was higher than that of the common antiviral drug ribavirin (20.44 + 4.64%) Indicating excellent efficacy (Table 3 and Fig. 5).

division
Antiviral activity (%) at a concentration of 100 / / ml Hepatitis A virus Hydrocortisone 57.55 + - 4.31 Ribavirin 20.44 + - 4.64

Claims (10)

An antiviral pharmaceutical composition comprising hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.
The antiviral pharmaceutical composition according to claim 1, wherein the virus is a virus belonging to the genus Picornaviridae .
The virus according to claim 2, wherein the virus belonging to the picornaviridae is selected from the group consisting of rhinovirus, enterovirus, cardiovirus, aphthovirus, poliovirus and hepatovirus wherein the antiviral composition is hepatovirus.
4. The antiviral pharmaceutical composition according to claim 3, wherein the rhinovirus is any one selected from the group consisting of rhinovirus type 2, type 3, and type 5.
The antiviral pharmaceutical composition according to claim 3, wherein the hepato virus is hepatitis A virus.
An antiviral health functional food containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.
The antifungal health food according to claim 6, wherein the virus is a virus belonging to the genus Picornaviridae.
The virus according to claim 7, wherein the virus belonging to the picornaviridae is selected from the group consisting of rhinovirus, enterovirus, cardiovirus, aphthovirus, poliovirus and hepatovirus wherein the hepatitis virus is hepatovirus.
A pharmaceutical composition for the prevention and treatment of viral diseases containing hydrocortisone or a pharmaceutically acceptable salt thereof as an active ingredient.
10. The method of claim 9, wherein the viral disease is selected from the group consisting of polio, paralysis, acute hemorrhagic conjunctivitis, viral meningitis, aquatic diseases, waterborne disease, hepatitis A, myositis, myocarditis, pancreatitis, diabetes mellitus, encephalitis, Wherein the pharmaceutical composition is a pharmaceutical composition for the prevention and treatment of viral diseases, characterized in that it is a salt thereof, a foot-and-mouth disease, asthma, chronic obstructive pulmonary disease, pneumonia, sinusitis or otitis media.

Images