KR102661593B1 - Pharmaceutical composition for preventing or treating severe fever with thrombocytopenia syndrome comprising of OSU-03012 as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome, comprising OSU-03012 (AR-12) as an active ingredient.

Description

Pharmaceutical composition for preventing or treating severe fever with thrombocytopenia syndrome comprising OSU-03012 as an active ingredient}

The present invention relates to a pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome containing OSU-03012 as an active ingredient.

Severe Fever with Thrombocytopenia Syndrome (SFTS) is a febrile hemorrhagic disease caused by the SFTS virus (SFTSV) and is a new, zoonotic infectious disease caused by ticks. In 2009, a number of patients with unexplained fever with high fever accompanied by a decrease in platelets, digestive symptoms, and leukopenia occurred in central and northeastern China. Through two years of epidemiological investigation and pathogen research, a single strand of infection was discovered from the patient's blood in 2011. An RNA virus was first reported as isolated SFTSV. Since then, cases of infection have been reported in China, Korea and Japan in 2012, and recently, cases have also been reported in Vietnam and Thailand. In severe cases, death may occur due to systemic inflammatory response syndrome and multiple organ failure, and the mortality rate is 6-30%.

The causative agent that causes SFTS is the official name of Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV), Dabie bandavirus , belonging to the Banyangvirus genus, order Bunyavirales , Phenuiviridae family, and is commonly called Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV). It is a spherical virus with a lipid membrane structure and has a genome consisting of three negative single-stranded RNA segments. The L-segment encodes RNA-dependent RNA polymerase (RdRp), and the M-segment encodes two glycoproteins (Gn/Gc). Nucleoproteins (NPs) and non-structural proteins (NSs) are expressed in the S-segment.

In Korea, the first case of infection was reported in 2012, and the number of confirmed cases is steadily increasing. In Korea, it was designated as a group 4 notifiable infectious disease in 2013 and surveillance of patient outbreaks began, and since 2020, it has been classified as a class 3 notifiable infectious disease, and outbreak and prevalence surveillance is being conducted. Infection is mainly caused by the bite of a tick infected with SFTSV, and occurs seasonally from February to November, mainly from spring to fall. Elderly people over 50 years old account for more than 90% of all confirmed cases, and the cumulative number of deaths from 2013 to 2021 is a total of 279, showing a fatality rate of 18.5%.

SFTS is mainly caused by the Haemaphysalis longicornis, and is known to be mainly distributed in China, Korea, Japan, Russia, and Australia, and was recently discovered in the United States. SFTS virus can not only infect humans, but also domestic animals such as cows, pigs, horses, and companion animals such as cats and dogs, and human infection can occur through these intermediate hosts. Additionally, it can be spread between people through contact with the body fluids or blood of an infected patient, or close contact with aerosols. As such, the risk of SFTS virus infection and the number of infected patients are increasing, but to date, there is no effective vaccine or treatment, so conservative treatment is mainly used, with methods such as ribavirin (antiviral treatment) administration, steroids, immunoglobulin administration, and plasma exchange. It is being attempted.

Several researchers have confirmed the antiviral efficacy against SFTSV in vitro or in vivo using existing FDA-approved compounds, nucleoside ananlogs, single antibodies, and antisera obtained from recovered SFTS patients. Favipiravir (T-705) is a guanosine analog that targets the RNA-dependent RNA polymerase of RNA viruses and inhibits viral replication. Currently, antiviral efficacy has been confirmed in interferon receptor-deficient mice (C57BL/6 IFNAR-/-), a SFTSV-susceptible animal model, and phase 3 clinical trials are in progress. The single antibody Ab10 targets the glycoprotein Gn, and its SFTSV antiviral efficacy was confirmed in interferon receptor-deficient mice (A129). However, the animal model that verified the in vivo efficacy is an immunosuppressed mouse, which does not reflect the complete human infection response, and favipiravir is not approved in Korea, making it difficult to use as a treatment for SFTS.

Korean Patent No. 2405350

In order to solve the above problems, the purpose of the present invention was to select the OSU-03012 compound that inhibits SFTS virus growth through a cell-based compound library screening method, and to confirm the antiviral efficacy, OSU-03012 was included as an active ingredient. The aim is to provide a pharmaceutical composition for preventing or treating severe fever with thrombocytopenia syndrome.

In order to solve the above problems, the present invention provides a composition for inhibiting severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a salt thereof as an active ingredient.

[Formula 1]

The compound represented by Formula 1 or its salt is administered in an amount of 0.001 to 10 g per day, and the composition may further include a component that has an inhibitory effect on the severe fever thrombocytopenia syndrome virus.

As an example, the present invention provides a pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is administered in an amount of 0.001 to 10 g per day, and the composition may further include an ingredient effective for severe febrile thrombocytopenia syndrome.

As another example, the present invention provides a food composition for preventing or improving severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a foodologically acceptable salt thereof as an active ingredient.

[Formula 1]

The compound represented by Formula 1 or a food-acceptable salt thereof is administered in an amount of 0.001 to 10 g per day, and the composition may further include an ingredient effective for severe febrile thrombocytopenia syndrome.

As another example, the present invention provides a method for inhibiting severe febrile thrombocytopenia syndrome virus in vitro, comprising administering a compound represented by the following formula (1) or a salt thereof.

[Formula 1]

As another example, the present invention provides a method for preventing or treating severe febrile thrombocytopenia syndrome in non-human animals, comprising administering a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

The pharmaceutical composition for the prevention or treatment of severe fever thrombocytopenia syndrome containing OSU-03012 of the present invention as an active ingredient not only provides new information necessary for follow-up research and development of anti-SFTS virus treatments, but also provides new opportunities for selection in the antiviral market. can be provided.

Figure 1 shows the antiviral efficacy of OSU-03012 according to the present invention against SFTS virus.

Hereinafter, preferred embodiments of the present invention will be described in detail. In addition, in the following description, many specific details, such as specific components, are shown, but this is provided to facilitate a more general understanding of the present invention, and it is known in the art that the present invention can be practiced without these specific details. It will be self-evident to those who have the knowledge. Additionally, in describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description will be omitted.

In the present invention, the term “prevention” or “prevention” means suppressing or delaying the occurrence of a disease from its cause.

In this specification, “treatment” means stopping the progression of damage by suppressing the progression and/or worsening of symptoms even without complete cure, or improving some or all of the symptoms and leading toward healing.

In the present invention, the term “improvement” refers to any action that improves or beneficially changes symptoms.

In order to achieve the object of the present invention, the present invention provides a composition for inhibiting severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a salt thereof as an active ingredient.

[Formula 1]

OSU-03012 (AR-12), a compound represented by Formula 1, is a compound having the structure of Formula 1 and has Cas No. It is a 742112-33-0 compound (IUPAC name: 2-Amino-N-[4-[5-phenanthren-2-yl-3-(trifluoromethyl)pyrazol-1-yl]phenyl]acetamide). Although it is a celecoxib derivative, it does not inhibit COX and is known to have antibacterial and antitumor uses as an inhibitor of various enzymes including PDK1.

The compound of the present invention may include a salt form, which can be prepared by methods common in the art, for example, inorganic salts such as hydrochloric acid, hydrogen bromide, sulfuric acid, sodium bisulfate, phosphoric acid, and carbonic acid. Salts of acids or with organic acids such as formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestisic acid, fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin). It means forming metal salts by reacting with alkali metal ions such as sodium or potassium, or forming another type of salt by reacting with ammonium ions.

The composition of the present invention can be manufactured in any form by further including appropriate carriers, excipients and diluents commonly used in the preparation of the composition, and is preferably manufactured in the form of a pharmaceutical composition or (health function) food composition. However, it is not limited to this.

As an example, the present invention provides a pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

The pharmaceutical composition of the present invention can be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions according to conventional methods. , tablets, capsules, injections and liquid formulations are more preferable. This formulation can be performed by a method commonly performed in the pharmaceutical field, and can be preferably formulated according to each disease or ingredient using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA.

Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, and cellulose. , methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

When formulated, it can be prepared by additionally using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose, or lactose ( It is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous agents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween, cacao, laurin, glycerogeratin, etc. can be used.

The pharmaceutical composition of the present invention can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method, and the preferred dosage is determined by the patient's condition and weight, and the severity of the disease. It varies depending on the degree, drug type, administration route and period, but can be appropriately selected by a person skilled in the art. However, for desirable effects, the pharmaceutical composition of the present invention may be included in an amount of 0.001 to 99.9% by weight, preferably 0.01 to 99% by weight, per day. The daily dosage may be about 0.01 to 1,000 mg/kg, preferably 100 to 300 mg/kg. If necessary, the active ingredient may be administered in an amount of 0.001 to 10 g per day, but is not limited thereto.

Additionally, in the composition according to one embodiment of the present invention, the composition may contain additional ingredients useful for inhibiting, preventing, treating or improving severe fever thrombocytopenia syndrome virus or severe fever thrombocytopenia syndrome, and the additional ingredients include compounds, It may contain all ingredients known to have effects including natural products.

As another example, the present invention provides a food composition for preventing or improving severe febrile thrombocytopenia syndrome, comprising a compound represented by the following formula (1) or a foodologically acceptable salt thereof as an active ingredient.

[Formula 1]

Food compositions to which the above composition can be added (health function) can be manufactured in the form of powders, granules, tablets, capsules or beverages. For example, there are various general foods, beverages, gum, tea, vitamin complexes, or health supplements.

The food composition of the present invention may include allitiamine, fursultiamine, benfotiamine, or salts thereof, as well as ingredients commonly added during food production, such as proteins, carbohydrates, fats, nutrients, and seasonings. Includes agents and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured as a drink, in addition to allitiamine, fursultiamine, benfotiamine or salts thereof, citric acid, high-fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, cephalic acid extract, jujube extract, and licorice. Extracts, etc. may additionally be included.

Additionally, the composition can be added to food or beverages for the purpose of preventing diseases. At this time, the amount of the active ingredient in the food or beverage can be added at 0.001 to 15% by weight of the total food weight, and the food composition can be added at a rate of 0.002 to 5 g, preferably 0.03 to 1 g, based on 100 g. and, if necessary, the active ingredient may be administered in an amount of 0.001 to 10 g per day, but is not limited thereto.

As another example, the present invention provides a method for inhibiting severe febrile thrombocytopenia syndrome virus in vitro, comprising administering a compound represented by the following formula (1) or a salt thereof.

[Formula 1]

As another example, the present invention provides a method for preventing or treating severe febrile thrombocytopenia syndrome in non-human animals, comprising administering a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

The 'administration' means introducing a predetermined substance into a cell or organism by an appropriate method, and in the present invention, the route of administration of the composition to an animal may be through any general route as long as it can reach the target tissue. For example, it may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, orally, topically, intranasally, intrapulmonaryly, or rectally, but is not limited thereto. However, when administered orally, proteins are digested, so it is desirable for oral compositions to be coated with the active agent or formulated to protect them from decomposition in the stomach. Additionally, the pharmaceutical composition can be administered by any device that allows the active agent to move to target cells.

The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms, and the present embodiments are merely intended to ensure that the disclosure of the present invention is complete and to include common knowledge in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the invention is only defined by the scope of the claims.

<Example 1> Preparation of materials

Vero E6 cells were purchased from ATCC (Manassas, USA) and incubated with 10% inactivated FBS (Fetal bovine serum, Gibco, USA) and 1% penicillin/streptomycin (Gibco, USA) in an incubator at 37°C with 5% _CO 2 . Cultured in supplemented Dulbecco Modified Eagle Medium (DMEM, Gibco, USA). The SFTS virus (passage 7, KADGH strain) was isolated from a domestic patient and was used under strict control in a biosafety level 3 (BSL-3) research facility. The virus was propagated in Vero E6 cells, and the virus titer was measured using an immunostaining experiment (Infectious center assay). The compound OSU-03012 used in the experiment was purchased in powder form from Chemscene (NJ, USA) and dissolved in DMSO solvent.

<Example 2> Virus infection and compound treatment

24 hours before the experiment, 0.4 x 10 ⁵ Vero E6 cells were distributed and cultured in each well of a 96-well plate. After moving the cells to a biosafety level 3 experimental area, the virus was infected with a multiplicity of infection (MOI) of 0.1 and treated with the compound to a final concentration of 10 μM. Immunostaining experiments were performed 24 hours later.

<Example 3> Immunostaining experiment (Infectious center assay)

After removing the cell culture medium of Vero E6 cells treated with viruses and compounds, 4% paraformaldehye was added and the cells were fixed for more than 5 minutes. After detection with a self-produced primary antibody against the SFTS virus nucleoprotein (NP), infection was observed using a HRP (horseradish peroxidase)-conjugated secondary antibody (Abfrontier, Korea). The antiviral efficacy of the compound was analyzed from the experimental results obtained by repeating the test twice.

<Experimental Example> Antiviral efficacy of OSU-03012 against SFTS virus

OSU-03012 shows that it inhibits SFTS virus proliferation (Figure 1). In the non-compound treated group, SFTSV nucleoprotein (NP) expression was well observed, whereas in the compound treated group, nuclear protein expression was decreased. Nuclear protein expression foci were measured and schematized.

Claims (11)

A composition for inhibiting severe fever thrombocytopenia syndrome virus comprising the compound OSU-03012 (AR-12) represented by the following formula (1) or a salt thereof as an active ingredient:
[Formula 1]
The composition for inhibiting severe febrile thrombocytopenia syndrome virus according to claim 1, wherein the compound OSU-03012 (AR-12) or a salt thereof represented by Formula 1 is administered in an amount of 0.001 to 10 g per day. The composition for inhibiting the severe febrile thrombocytopenia syndrome virus according to claim 1, wherein the composition further comprises an ingredient that has an inhibitory effect on the severe febrile thrombocytopenia syndrome virus. A pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome, comprising the compound OSU-03012 (AR-12) represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]
The method of claim 4, wherein the compound OSU-03012 (AR-12) represented by Formula 1 or a pharmaceutically acceptable salt thereof is administered for the prevention or treatment of severe febrile thrombocytopenia syndrome, characterized in that 0.001 to 10 g per day. pharmaceutical composition The pharmaceutical composition for preventing or treating severe febrile thrombocytopenia syndrome according to claim 4, wherein the composition further comprises an ingredient effective for severe febrile thrombocytopenia syndrome. A food composition for preventing or improving severe febrile thrombocytopenia syndrome comprising the compound OSU-03012 (AR-12) represented by the following formula (1) or a food-acceptable salt thereof as an active ingredient:
[Formula 1]
The method of claim 7, wherein the compound OSU-03012 (AR-12) represented by Formula 1 or a food-acceptable salt thereof is administered for the prevention or improvement of severe febrile thrombocytopenia syndrome, characterized in that 0.001 to 10 g per day. food composition The food composition for preventing or improving severe febrile thrombocytopenia syndrome according to claim 7, wherein the composition further comprises ingredients effective for severe febrile thrombocytopenia syndrome. A method of inhibiting severe febrile thrombocytopenia syndrome virus in vitro comprising administering the compound OSU-03012 (AR-12) or a salt thereof represented by the following formula (1):
[Formula 1]
A method for preventing or treating severe febrile thrombocytopenia syndrome in non-human animals comprising administering the compound OSU-03012 (AR-12) represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]