KR20240018822A - Composition for preventing or treating coronavirus infection or disease caused by coronavirus infection comprising gossypol - Google Patents

Abstract

The present invention uses gossypol or gossypol derivatives to effectively prevent or treat infections caused by coronaviruses or diseases caused by coronaviruses, as well as disinfection or cleaning against coronaviruses, through the coronavirus inhibitory activity of these compounds. You can.

Description

Composition for preventing or treating coronavirus infection or disease caused by coronavirus infection comprising gossypol}

The present invention relates to a composition that can effectively prevent or treat coronavirus infections or diseases caused by such infections, as well as compositions for disinfection or cleaning.

Coronavirus is a representative virus that causes fatal infectious diseases in modern civilization. In April 2003, Severe Acute Respiratory Syndrome (SARS), also known as SARS, broke out and killed many people with a mortality rate of 9.6%. As Middle East Respiratory Syndrome (MERS), also known as MERS, spread from the Middle East to the rest of the world, there were many deaths with a mortality rate of about 36%. In addition, coronavirus disease 19 (Coronavirus disease 19; COVID-19) is a new type of coronavirus that first occurred in December 2019 and spread worldwide. Severe acute respiratory syndrome coronavirus 2 (SARS) It is a respiratory infectious disease caused by -CoV-2). Coronavirus infection-19 is transmitted when droplets from an infected person penetrate the respiratory tract or mucous membranes of the eyes, nose, and mouth. If infected, after an incubation period of about 2 to 14 days (estimated), fever (37.5 degrees), respiratory symptoms such as coughing or difficulty breathing, and pneumonia appear as main symptoms, and the fatality rate is 5.6% according to data collected as of March 2020. It reaches.

The epidemic of new variants of the virus is causing great problems for humanity, but although the development of treatments for coronaviruses is being promoted so far, compositions for preventing or treating infected people in medical staff and contacts at risk of infection have not been completed. . Therefore, a composition that can effectively prevent, disinfect, or treat coronavirus infection or infection is required.

One object of the present invention is to provide a composition for various uses that can prevent infection by coronavirus or various diseases caused by the infection.

Another object of the present invention is to provide a preventive or therapeutic composition having inhibitory activity against coronavirus.

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

DETAILED DESCRIPTION OF THE INVENTION Various embodiments described herein are described below with reference to the drawings. In the following description, various specific details, such as specific forms, compositions, and processes, are set forth in order to provide a thorough understanding of the invention. However, certain embodiments may be practiced without one or more of these specific details or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques are not described in specific detail so as not to unnecessarily obscure the invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Accordingly, the phrases “in one embodiment” or “an embodiment” expressed in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, particular features, shapes, compositions, or properties may be combined in any suitable way in one or more embodiments.

Unless there is a special definition within the present invention, all scientific and technical terms used in this specification have the same meaning as commonly understood by a person skilled in the art in the technical field to which the present invention pertains.

One embodiment of the present invention relates to a composition for preventing or treating viral infection or infectious disease, comprising gossypol as an active ingredient.

According to another embodiment of the present invention, the subject is given the gossypol; or a method for preventing viral infection or infectious disease, comprising administering a pharmaceutically acceptable salt thereof.

The "gossypol" of the present invention is a type of polyphenol compound contained in the separate pigment glands of seeds, leaves, stems, and roots of plants of the genus Gossypium and parts of Malvaceae plants. It is also called polyphenolic gossypol or cottonseed pigment. It is expressed by the formula 1 below. Provides plants with resistance to pests. When gossypol is added to poultry feed, reductions in feed availability, egg productivity, and yolk discoloration of stored eggs have been reported. On the other hand, ruminants deactivate gossypol through fermentation. Free gossypol is physiologically toxic, whereas bound gossypol is inactive. Non-protein components of cottonseed also combine with gossypol to form insoluble and/or indigestible complexes. Although this combination detoxifies the gossypol in cottonseed meal, it reduces its protein and biological value. Adding iron at a ratio of 2:1 or 3:1 to free gossypol can effectively reduce the toxicity of gossypol in the liver. In China, gossypol was discovered to suppress male sperm function and is being studied as a male oral contraceptive. In the present invention, gossypol includes all derivatives of gossypol such as (+)-gossypol, (-)-gossypol, or their linked forms (±)-gossypol, hemigossypol, and apogossypol. It means.

In the present invention, the gossypol derivative is not limited thereto, but is preferably (+)-gossypol.

Said pharmaceutically acceptable salts in the present invention may be salts generally considered by those skilled in the art to be suitable for medical applications (e.g. because such salts are not harmful to the subjects to be treated with said salts), or, respectively. It is a salt that causes acceptable side effects within the treatment of. Generally, the pharmaceutically acceptable salts are salts that are considered acceptable by regulatory authorities, such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or the Pharmaceutical and Medical Device Agency (PMDA) of the Japanese Ministry of Health, Labor and Welfare. . However, the present invention in principle also provides, for example, intermediates in the preparation of the compounds according to the invention or physiologically functional derivatives thereof, or pharmaceutically acceptable salts of the compounds according to the invention or physiologically functional derivatives thereof. As intermediates in the preparation of derivatives, also include salts of the compounds according to the invention which are not pharmaceutically acceptable as such. The salts include water-insoluble salts and, in particular, water-soluble salts.

In each case, the person skilled in the art will be able to determine whether a particular compound according to the invention or a physiologically functional derivative thereof is capable of forming salts, i.e. whether the compound according to the invention or a physiologically functional derivative thereof is capable of forming salts, e.g. For example, it is easy to determine whether it has a group that can bear a charge, such as an amino group, carboxylic acid group, etc.

Exemplary salts of the compounds of the invention are acid addition salts or salts with bases, especially pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases commonly used in pharmaceuticals, which are water-insoluble or especially water-soluble acid addition salts. It's salt. Depending on the substituents of the compounds of the invention, salts with bases may also be suitable. Acid addition salts include, for example, a solution of a compound of the invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. It can be formed by mixing. Likewise, pharmaceutically acceptable base addition salts include alkali metal salts (eg, sodium or potassium salts); alkaline earth metal salts (eg, calcium or magnesium salts); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium and amines formed using counter anions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl sulfonates and aryl sulfonates). cations). Illustrative examples of pharmaceutically acceptable salts include acetate, adipate, alginate, arginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, Bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, digluconate, dihydrochloride, dodecyl sulfate, edetate, edisylate, ethanesulfonate, Formate, fumarate, galactate, galacturonate, gluconate, glutamate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrobromide, hydrochloride, hydroiodide. , 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide, isobutyrate, isothionate, lactate, laurate, lauryl sulfate, maleate, maleate, malonate, mandelate, methane. Sulfonate (mesylate), methyl sulfate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate. /diphosphate, phthalate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, suberate, succinate, tannate, tartrate, tosylate, undecanoate, Includes, but is not limited to, valerate.

Salts which are not pharmaceutically acceptable in the present invention and which can be obtained, for example, as process products during the production of the compounds according to the invention on an industrial scale, are also included in the present invention and, if desired, are known to the person skilled in the art. It can be converted into a pharmaceutically acceptable salt by a method.

In the present invention, the virus may be an RNA virus. In the present invention, the “RNA virus” refers to all viruses that use RNA as genetic material. For example, the RNA viruses include Coronaviridae, Amalgaviridae, Birnaviridae, Chrysoviridae, Cystoviridae, and Endornaviridae ( Endornaviridae, Hypoviridae, Megabirnaviridae, Partitiviridae, Picobirnaviridae, Reoviridae, Totiviridae, Quadriviridae, Arteriviridae, Mesoniviridae, Roniviridae, Dicistroviridae, Iflaviridae, Marnaviridae Marnaviridae, Picornaviridae, Secoviridae, Alphaflexiviridae, Betaflexiviridae, Gammaflexiviridae, Tymoviridae ), Bornnaviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Nyamiviridae, Caliciviridae, Pla Flaviviridae, Luteoviridae, Togaviridae, Pneumoviridae, Arenaviridae, Deltavirus, or Orthomyxviridae ( Orthomyxoviridae) virus, preferably Coronaviridae, and more preferably a coronavirus belonging to Coronaviridae.

In the present invention, the "Coronavirus" is an RNA virus with a gene size of 27 to 32 kb, which has four genera (alpha, beta, gamma, and delta) in the Coronavirinae subfamily of Coronaviridae. It is known to cause respiratory and digestive system infections in humans and animals. It is mainly transmitted through mucous membranes and droplets. In humans, it generally causes mild respiratory infections, but in rare cases, it can cause fatal infections. In cows and pigs, it can cause diarrhea, and in chickens, it can cause respiratory diseases. Among coronaviruses, coronaviruses that host humans are divided into the categories shown in Table 1 below (Korea Centers for Disease Control and Prevention, 2020). Among the four genera, Alpha and Beta are reported to infect humans and animals, while Gamma and Delta are reported to infect only animals.

genus People - Coronavirus non-human coronavirus alpha coronavirus 229, NL63 Porcine epidemic diarrhea virus (PEDV), (porcine) transmissible gastroenteritis virus (TGEV), canine coronavirus (CCoV), feline coronavirus (FCoV), Miniopterus bat ( Bat) coronavirus1, Miniopterus bat coronavirus HKU8, Rhinolophus bat coronavirus HKU2, Scotophilus bat coronavirus 512 beta coronavirus OC43, HKU1, SARS-CoV, SARS-CoV2, MERS-CoV Porcine hemagglutinating encephalomyelitis virus (PHEV), bovine coronavirus (BCoV), equine coronavirus (EqCoV), murine coronavirus (MuCoV), Tylonycteris bat coronavirus HKU4, Pipistrellus bat coronavirus HKU5, Rousettus bat coronavirus HKU9 gamma coronavirus - Avian coronavirus, Beluga whale-Coronavirus SW1 delta coronavirus - Jeju Jigbakguri (Bulbul)-Coronavirus HKU11, Thrush-Coronavirus HKU12, Kinbara (Munia)-Coronavirus HKU13

In particular, there are currently seven types of coronaviruses that are infectious to humans, as shown in Table 2 below, including the type that causes colds (229E, OC43, NL63, HKU1) and the type that causes severe pneumonia (SARS-CoV, SARS-CoV2, MERS). -CoV).

Virus name Genus host Symptom HCoV-229E Alpha Human mild respiratory symptoms HCoV-NL63 Alpha Human mild respiratory symptoms SARS-CoV Beta Human severe respiratory symptoms MERS-CoV Beta Human severe respiratory symptoms HCoV-OC43 Beta Human mild respiratory symptoms HCoV-HKU1 Beta Human pneumonia symptoms SARS-CoV-2 Beta Human Mild respiratory symptoms and in severe cases, difficulty breathing

In the present invention, the "Beta coronavirus" is one of the four coronaviruses of the coronavirus subfamily and corresponds to a zoonotic infection. Examples of beta coronaviruses include Severe Acute Respiratory Syndrome virus (SARS; SARS-CoV), Severe Acute Respiratory Syndrome virus-2 (SARS-CoV-2), and Middle East Respiratory Syndrome virus (SARS-CoV). Syndrome viruses (Middle East Respiratory Syndrome virus; MERS; MERS-CoV), human coronavirus OC43 (HCoV-OC43), or human coronavirus HKU1 (HCoV-HKU1) are known to exist.

In the present invention, the "Severe Acute Respiratory Syndrome virus-2 (SARS-CoV-2)" corresponds to an enveloped, positive-polar single-stranded RNA beta coronavirus belonging to the Coronaviridae family. . Coronaviruses that historically infect humans are severe common cold viruses, including hCoV-OC43, HKU, and 229E5. Analyzing the sequence of the SARS-CoV-2 isolate, the 30-kb genome is known to encode 14 open-reading frames (ORFs), and 13 ORFs at the 3' end of the viral genome are 9. It is expressed from predicted sub-genomic RNA, which consists of four structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N), and 9 It is expressed from two putative additional elements. Here, the nucleocapsid protein includes N1, N2, and N3 segments.

In the present invention, the coronavirus includes naturally or artificially mutated variants.

In the present invention, the infectious disease caused by the coronavirus is coronavirus enteritis, coronavirus diarrhea, severe acute respiratory syndrome coronavirus (SARS), Middle East respiratory syndrome (MERS), or these It may be a combination of, but any disease that can be caused by the coronavirus infection may be included without limitation.

In the present invention, the infectious disease may be a respiratory disease, hepatitis, gastroenteritis, or encephalitis caused by the viral infection.

In the present invention, the respiratory disease may cause Severe Acute Respiratory Syndrome (SARS). The severe acute respiratory syndrome is a disease that can cause upper or lower respiratory tract infection, causing sudden fever, cough, and difficulty breathing, and can progress to pneumonia and lead to death.

The composition of the present invention can be used alone, in combination with conventional antiviral agents, or in combination with biological agents such as antiserum.

The composition of the present invention may be used as a pharmaceutical composition, food composition, food additive composition, feed composition, or feed additive composition, but is not particularly limited.

In the present invention, “prevention” refers to any action that delays the progression of viral infection or proliferation by administering the composition of the present invention.

In the present invention, the gossypol or pharmaceutical composition may be in the form of a capsule, tablet, granule, injection, ointment, powder, or beverage, and the pharmaceutical composition may be used for human subjects. .

The pharmaceutical composition of the present invention is not limited to these, but can be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, external preparations, suppositories, and sterile injection solutions according to conventional methods. You can. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. for oral administration, and buffers, preservatives, and analgesics for injections. Topics, solubilizers, isotonic agents, stabilizers, etc. can be mixed and used, and for topical administration, bases, excipients, lubricants, preservatives, etc. can be used. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing it with a pharmaceutically acceptable carrier as described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injections, it can be manufactured in the form of unit dosage ampoules or multiple dosage forms. there is. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release preparation, etc.

Meanwhile, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil may be used. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be additionally included.

The route of administration of the compound or pharmaceutical composition according to the present invention is not limited to these, but is oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral. , topical, sublingual, or rectal. Oral or parenteral administration is preferred.

As used herein, “parenteral” includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention can also be administered in the form of a suppository for rectal administration.

The compounds or pharmaceutical compositions of the present invention may be influenced by several factors, including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, excretion rate, drug formulation, and the severity of the particular disease to be prevented or treated. The dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but may be appropriately selected by a person skilled in the art, and may range from 0.0001 to 0.0001 per day. It can be administered at 50 mg/kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered several times. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

In the present invention, the "individual" refers to an object that has symptoms or is suspected of being infected with a virus and needs to prevent a viral infection or a disease caused by the infection by suppressing viral activity, etc., and is already infected or may be infected with the virus. This means all animals, including humans.

The "administration" of the present invention refers to the process of introducing the active ingredient of the present invention into an individual by any appropriate method. In the treatment method of the present invention, the administration method is through various routes such as oral or parenteral. may be administered.

For the purpose of the present invention, the specific pharmaceutically effective amount for the subject of interest is the type and degree of response to be achieved, the composition containing the active ingredient, the patient's age, and whether other agents are used as the case may be. Various factors well known in the medical field, including body weight, general health condition, gender and diet, administration time, administration route and secretion rate of the composition containing the active ingredient, treatment period, and drugs used together or simultaneously with the specific composition. It is desirable to apply it differently depending on similar factors.

In the present invention, the food composition can be manufactured in the form of various foods, such as beverages, gum, tea, vitamin complexes, powders, granules, tablets, capsules, cookies, rice cakes, bread, etc. It can also be used as a food additive composition necessary for manufacturing the food composition.

When gossypol, which is included as an active ingredient in the present invention, is included in a food composition, it can be added in an amount of 0.1 to 50% of the total weight. Here, when the food composition is manufactured in the form of a beverage, there are no particular limitations other than containing the food composition in the indicated ratio, and various flavoring agents or natural carbohydrates can be contained as additional ingredients like ordinary beverages. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. can do. Examples of the flavoring agent include natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.).

Other food compositions or food composition additives of the present invention include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, It may contain protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

These ingredients can be used independently or in combination. The proportion of these additives is not that critical, but is generally selected in the range of 0.1 to about 50 parts by weight per 100 parts by weight of the food composition of the present invention.

When the gossypol of the present invention is contained and used in a feed composition or feed additive composition, the composition may be a highly concentrated solution of 20 to 90% or may be prepared in the form of powder or granules. The feed additives include organic acids such as citric acid, malic acid, adipic acid, lactic acid, and malic acid, phosphates such as sodium phosphate, potassium phosphate, and acid pyrophosphate, polyphenols, catechin, alpha-tocopherol, rosemary extract, vitamin C, and green tea extract. , licorice extract, chitosan, tannic acid, phytic acid, etc., may additionally contain one or more of natural antioxidants.

In the present invention, when the gossypol is used as a feed, the composition can be formulated in the form of a normal feed and may include common feed ingredients. The feed additives and feeds include grains such as ground or crushed wheat, oats, barley, corn and rice; Vegetable protein feeds, such as those based on rape, soybean and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; It may further include dry ingredients such as sugar and dairy products, such as various powdered milk and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, etc.

In the present invention, when gossypol is used as the feed additive, it can be administered to animals alone or in combination with other feed additives in an edible carrier. Additionally, the feed additives can be easily administered to animals as a top dressing, by mixing them directly into animal feed, or in an oral formulation separate from the feed. When the feed additive is administered separately from animal feed, it can be prepared into an immediate-release or sustained-release formulation by combining it with a pharmaceutically acceptable edible carrier, as is well known in the art. These edible carriers can be solid or liquid, such as corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the feed additive may be a tablet, capsule, powder, troche or sugar-containing tablet or a top dressing in microdisperse form. If a liquid carrier is used, the feed additive may be in the form of gelatin soft capsules, or in the form of syrup, suspension, emulsion, or solution. The feed may include any protein-containing organic cereal flour commonly used to meet the dietary needs of animals. These protein-containing flours typically consist of corn, soybean flour, or corn/soybean flour mix. Additionally, the feed composition or feed additive composition may contain, for example, preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, etc. Additionally, the feed additive composition can be used by adding it to animal feed by dipping, spraying, or mixing.

According to another embodiment of the present invention, gossypol; or a pharmaceutically acceptable salt thereof as an active ingredient, and relates to a disinfecting or cleaning composition having inhibitory activity against viruses.

According to another embodiment of the present invention, the subject is provided with the gossypol; or a pharmaceutically acceptable salt thereof.

In the present invention, the definitions of gossypol, virus, and pharmaceutically acceptable salts overlap with those described above and are omitted hereinafter to prevent excessive complexity of the specification.

In the present invention, “disinfection” means losing the activity of or killing pathogenic microorganisms or pathogenic viruses. Deactivating includes inhibiting the infection, survival, proliferation, spread, or combinations of pathogenic microorganisms or pathogenic viruses.

As used herein, “cleaning” refers to methods used to facilitate or assist in removing contaminants, bleaching, reducing microbial or viral populations, rinsing, and any combination thereof.

In the present invention, the “object” includes all living or non-living objects (tools, devices, clothing, dishes, etc.) or their hard surfaces that must prevent viral infection.

Since the gossypol of the present invention has a specific virus-killing ability, the disinfecting composition or cleaning composition containing the above compound of the present invention is useful as a disinfectant and cleaner for hospital and health care to prevent hospital infections. It can be used for disinfection and cleaning of various items such as general household disinfectants and cleaners, tableware, food cooking equipment, building interiors, drinking water, vehicles, aircraft, and ships.

When the composition of the present invention is used as a disinfecting composition or cleaning composition, it can of course be included along with a pharmacologically, veterinarily, or agriculturally acceptable carrier or diluent.

In addition, the compound or composition of the present invention can be prepared in any of the liquid phase, solid phase, and gas phase.

The compound or composition of the present invention can be applied to the human body itself or various instruments or devices used by humans, and when applied to the human body, it can be administered in any manner such as orally or parenterally. In addition, the compound or composition can be formulated in various forms that can be effectively applied to the human body itself or various instruments or devices, for example, pharmaceutical formulations for oral administration such as tablets, pills, capsules, powder formulations, or sprays. , Cleansing L formulations applicable to external preparations such as gels, liquids, creams, ointments, etc., or instruments and devices, preferably sprays, but are not limited thereto.

In the present invention, as a method of applying the compound or composition to an object, the "contact" refers to a method of applying a known disinfection or cleaning agent, appropriately selected according to the nature of the object, surrounding environment, etc., such as wiping, dipping, or immersion. It may include immersing or spraying, preferably in the form of spraying, but is not limited thereto.

The composition provided by the present invention has coronavirus inhibitory activity, not only has a disinfection or cleaning effect against coronavirus, but can also effectively prevent or treat infection by coronavirus or diseases caused by the infection.

Figure 1 is a schematic diagram of a test confirming the effect of gossypol against coronavirus according to an embodiment of the present invention.
Figure 2 is a test result confirming the effect of gossypol on coronavirus according to an embodiment of the present invention.
Figure 3 shows test results confirming the effect of gossypol combination treatment on patient-derived temozolomide-resistant cells according to an embodiment of the present invention.

Hereinafter, the present invention will be explained in detail by the following examples. However, the following examples only illustrate the present invention, and the content of the present invention is not limited by the following examples.

Example

[Example 1] Confirmation of the effect of gossypol on coronavirus

Vero E6 cells (600000 cells) were plated in T25 flasks and pretreated with gossypol (5, 10, or 20 microMolar) and 0.2% DMSO as a negative control. Gossypol was used in two types, (+)-gossypol and (-)-gossypol. One hour after cell seeding, pretreated Vero E6 cells were infected with 0.3 MOI of SARS-CoV-2 virus particles belonging to the B.1.1.7 lineage (or UK variant). Uninfected cells were used as negative controls. After 72 hours, cells were lysed with TRIZOL and total RNA was extracted. The N1 gene fragment was measured via qPCR analysis using the SYBRGREEN approach (QuantStudio 5 machine) (Figure 1).

As a result of the experiment, the data showed that all cells pretreated with gossypol (P<0.05) had a statistically significant infection inhibition effect on viral infection compared to vehicle control infected cells. Specifically, when (+)-gossypol was pretreated, treatment with 5 microMolar gossypol had a better infection inhibition effect than treatment with 20 microMolar gossypol (P<0.05). In addition, in all comparisons of gossypol administration at the same concentration, (+)-gossypol had a superior coronavirus infection inhibition effect than (-)-gossypol (Figure 2) (P<0.05).

[Example 2] Confirmation of the effect of gossypol combination treatment on patient-derived temozolomide-resistant cells

(1) Isolation of glioblastoma cells

Patients whose brain tumor recurred even after receiving standard chemotherapy (Concurrent chemoradation therapy (CCRT) according to the Stupp protocol: 6 weeks of radiation therapy and Temozolomide drug treatment followed by 4 weeks of rest, followed by 6-week cycles of Temozolomide drug treatment) after glioblastoma brain tumor surgery Cells were isolated from the tissues. For this purpose, first, a part of the brain tumor tissue was removed, and then the tissue was cut to about 3-4 mm, or the tissue was minced in singel cell isolation mode using a C-tube. Afterwards, papain was treated and enzyme digestion was performed at 37°C for 30 minutes. This was dissolved in DMEM+FBS+antibiotics media and cultured for 4-5 days, and when the cell density reached 80-90%, positive selection was performed by attaching beads using a Tumor cell isolation kit (miltenyl biotec).

(2) Cell viability test

4000 selected cells were seeded and treated with drugs after 24 hours (n=3). Treatment drugs were prepared by adding 10 μl of drug to 90 μl of culture medium. Temozolomide (TMZ) was treated at a concentration of 0.1uM to 100uM, and gossypol (G) was treated alone or at a concentration of 0.1uM to 100uM. The TMZ+Gossypol group was treated at a concentration of 20mg/kg in accordance with the concentration of the oral feeding (in vivo) experiment, which is approximately 34uM when calculated. Cell death was observed 6 hours after the cells were treated with the drug, and cell survival was measured 72 hours later using Cell counting kit-8 (CK04-13). As a result, the group treated only with TMZ showed that cells survived well without death up to the maximum drug treatment concentration, and the group treated with TMZ+G20mg/kg showed cell survival rates at all concentrations except control (no drug treat). decreased. In the group treated with gossypol alone, cell survival began to decrease above drug concentration log3 (1uM) (Figure 3). From the above results, it was found that gossypol not only has an excellent therapeutic effect on glioblastoma as a single drug, but also has an excellent combined effect with TMZ.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (16)

A pharmaceutical composition for preventing or treating viral infections or infectious diseases, comprising gossypol or a pharmaceutically acceptable salt thereof as an active ingredient.
According to clause 1,
The gossypol is composed of (+)-gossypol, (-)-gossypol, (±)-gossypol, hemigosypol, and apogossypol, which are linked forms of (+)-gossypol and (-)-gossypol. A pharmaceutical composition selected from the group consisting of:
According to clause 2,
A pharmaceutical composition wherein the gossypol is (+)-gossypol.
According to clause 1,
A pharmaceutical composition, wherein the virus is a coronavirus.
According to clause 4,
The coronaviruses include human coronavirus 229E (HCoV-229E), human coronavirus OC43 (HCoV-OC43), human coronavirus HKU1 (HCoV-HKU1), human coronavirus NL63 (HCoV-NL63), and severe acute respiratory syndrome coronavirus. (SARS-CoV), Severe Acute Respiratory Syndrome virus-2 (SARS-CoV-2), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), porcine epidemic diarrhea virus; PEDV), transmissible gastroenteritis virus (TGEV), porcine hemagglutinating encephalomyelitis virus (PHEV), bovine coronavirus (BCoV), equine coronavirus (EqCoV), mouse coronavirus Viruses (murine coronavirus; MuCoV), canine coronavirus (CCoV), feline coronavirus (FCoV), bat coronavirus-1 (Miniopterus bat coronavirus1), bat coronavirus HKU8 (Miniopterus bat coronavirus HKU8), Bat coronavirus HKU2 (Rhinolophus bat coronavirus HKU2), bat coronavirus 512 (Scotophilus bat coronavirus 512), bat coronavirus HKU4 (Tylonycteris bat coronavirus HKU4), bat coronavirus HKU5 (Pipistrellus bat coronavirus HKU5), bat coronavirus HKU9 (Rousettus) bat coronavirus HKU9), Avian coronavirus, Beluga whale coronavirus SW1, Bulbul coronavirus HKU11, Thrush coronavirus HKU12, and Kinvara coronavirus. A pharmaceutical composition comprising at least one selected from the group consisting of the virus HKU13 (Munia coronavirus HKU13).
A food or food additive composition for preventing or improving viral infection or infectious disease, comprising gossypol or a pharmaceutically acceptable salt thereof as an active ingredient.
According to clause 6,
The gossypol is composed of (+)-gossypol, (-)-gossypol, (±)-gossypol, hemigosypol, and apogossypol, which are linked forms of (+)-gossypol and (-)-gossypol. A food or food additive composition selected from the group consisting of:
According to clause 6,
A food or food additive composition, wherein the virus is a coronavirus.
A feed or feed additive composition for preventing or improving viral infection or infectious disease, comprising gossypol or a pharmaceutically acceptable salt thereof as an active ingredient.
According to clause 9,
The gossypol is composed of (+)-gossypol, (-)-gossypol, (±)-gossypol, hemigosypol, and apogossypol, which are linked forms of (+)-gossypol and (-)-gossypol. A feed or feed additive composition selected from the group consisting of:
According to clause 9,
The feed or feed additive composition, wherein the virus is a coronavirus.
A disinfecting or cleaning composition having virus inhibitory activity, comprising gossypol or a pharmaceutically acceptable salt thereof as an active ingredient.
According to clause 12,
The gossypol is composed of (+)-gossypol, (-)-gossypol, (±)-gossypol, hemigosypol, and apogossypol, which are linked forms of (+)-gossypol and (-)-gossypol. A disinfecting or cleaning composition selected from the group consisting of:
According to clause 12,
A composition for disinfection or cleaning, wherein the virus is a coronavirus.
A method for preventing viral infection or infectious disease, comprising administering gossypol (polyphosphate) or a pharmaceutically acceptable salt thereof to an entity other than a human.
A method for disinfecting or cleaning a virus, comprising contacting a subject other than a human with gossypol or a pharmaceutically acceptable salt thereof.