[go: up one dir, main page]

WO2024205087A1 - Composition pharmaceutique pour la prévention ou le traitement d'infections virales, comprenant de la fucoxanthine - Google Patents

Composition pharmaceutique pour la prévention ou le traitement d'infections virales, comprenant de la fucoxanthine Download PDF

Info

Publication number
WO2024205087A1
WO2024205087A1 PCT/KR2024/003350 KR2024003350W WO2024205087A1 WO 2024205087 A1 WO2024205087 A1 WO 2024205087A1 KR 2024003350 W KR2024003350 W KR 2024003350W WO 2024205087 A1 WO2024205087 A1 WO 2024205087A1
Authority
WO
WIPO (PCT)
Prior art keywords
fucoxanthin
coronavirus
present
preventing
cov
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/KR2024/003350
Other languages
English (en)
Korean (ko)
Inventor
허수진
강나래
허성영
김은아
박아름이
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Korea Institute of Ocean Science and Technology KIOST
Original Assignee
Korea Institute of Ocean Science and Technology KIOST
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Korea Institute of Ocean Science and Technology KIOST filed Critical Korea Institute of Ocean Science and Technology KIOST
Publication of WO2024205087A1 publication Critical patent/WO2024205087A1/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/121—Heterocyclic compounds containing oxygen or sulfur as hetero atom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/336—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having three-membered rings, e.g. oxirane, fumagillin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/02—Algae
    • A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a pharmaceutical composition for preventing or treating viral infections, comprising fucoxanthin or a pharmaceutically acceptable salt thereof.
  • Coronaviruses are RNA viruses with a wide host range, including humans, birds, rodents, and mammals, and their genomes are about 30 kb in size, making them the largest of all RNA viruses. They are named coronavirus because the protein protrusions on the surface of the virus resemble a colona or crown.
  • coronaviruses There are seven known coronaviruses that can infect humans and cause disease. Four of them (HCoV-229E, HCoV-OC43, HCoV-NL63, and HKU1) are also called community-acquired respiratory HCoV (CAR HCoV). They occur more frequently in winter and spring than in summer and fall around the world in terms of temperature and climate, and are known to cause 10-30% of upper respiratory tract infections in adults.
  • coronaviruses SARS-CoV and MERS-CoV
  • SARS-CoV and MERS-CoV coronaviruses that caused the Severe Acute Respiratory Syndrome (SARS) epidemic in 2003 and the Middle East Respiratory Syndrome (MERS) epidemic in 2012 are species that are known to cause severe respiratory syndrome through lower respiratory tract infection, and are transmitted from animals to humans.
  • SARS-CoV-2 is a betacoronavirus strain that originated from bats, like the SARS and MERS coronaviruses. Analysis of the virus's genetic sequence confirmed that it has the highest homology (89.1%) with similar coronaviruses from bats. As COVID-19 spread worldwide, the World Health Organization (WHO) declared a pandemic on March 11, 2020, and COVID-19 has been rapidly spreading since then, making the development of a therapeutic that can treat or prevent COVID-19 urgent.
  • WHO World Health Organization
  • Zika virus is one of the viruses transmitted by mosquitoes, and is a single positive-stranded RNA virus belonging to the Flaviviridae family, a subgenus of the Flaviviridae genus, and is an infectious disease that causes congenital malformations in fetuses and newborns, such as microcephaly and neurological complications, such as Guillain-Barre syndrome.
  • Zika virus When Zika virus does cause disease in humans, it usually results in a self-limited febrile illness, characterized by low-grade fever, rash, headache, conjunctivitis, muscle and joint pain. More recently, Zika virus infection has been linked to an increased incidence of microcephaly in fetuses and infants, and Guillain-Barré syndrome in adults. The incubation period for Zika virus disease is not known exactly, but is thought to be several days to a week, with symptoms that are generally mild and lasting from several days to a week. Zika virus typically remains in the blood of an infected person for about a week, but may remain in some people for longer periods.
  • the inventors of the present invention studied a therapeutic agent capable of treating coronavirus infection including COVID-19 and Zika virus infection, and confirmed that fucoxanthin, an edible carotenoid, has excellent therapeutic efficacy against viral infections, thereby completing the present invention.
  • the purpose of the present invention is to provide a composition for preventing, improving or treating viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • Another object of the present invention is to provide a method for preventing or treating a viral infection, comprising a step of administering fucoxanthin or a pharmaceutically acceptable salt thereof to a subject in need thereof.
  • the present invention provides a pharmaceutical composition for preventing or treating viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the above fucoxanthin may be derived from marine brown algae.
  • the above brown algae may be Eisenia bicyclis, Sargassum siliquastrum or Hijikia fusiformis.
  • the above virus could be a coronavirus or a Zika virus.
  • the above coronavirus may be at least one selected from the group consisting of human coronavirus 229E (HCoV-229E), human coronavirus OC43 (HCoV-OC43), severe acute respiratory syndrome coronavirus (SARS-CoV), human coronavirus NL63 (HCoV-NL63, New Haven coronavirus), human coronavirus HKU1, Middle East respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2), and variants thereof.
  • HKU1 Middle East respiratory syndrome coronavirus
  • MERS-CoV Middle East respiratory syndrome coronavirus
  • SARS-Cov-2 severe acute respiratory syndrome coronavirus
  • the above SARS-Cov-2 variant may be at least one selected from the group consisting of alpha ( ⁇ ), beta ( ⁇ ), gamma ( ⁇ ), delta ( ⁇ ), and omicron ( ⁇ ) variants.
  • the concentration of the above fucoxanthin or a pharmaceutically acceptable salt thereof may be 0.1 to 100 ⁇ M.
  • the present invention provides a health functional food composition for preventing or improving viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a feed composition for preventing or improving viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a method for preventing or treating a viral infection, comprising a step of administering fucoxanthin or a pharmaceutically acceptable salt thereof to a subject in need thereof.
  • composition of the present invention for preventing, improving or treating a viral infection has excellent inhibitory activity against a coronavirus, and thus can be usefully applied to the treatment of a coronavirus infection such as COVID-19.
  • composition of the present invention for preventing, improving or treating a viral infection has excellent inhibitory activity against Zika virus, and thus can be usefully applied to the treatment of Zika virus infection.
  • Figure 1a shows the results of analyzing the cytotoxicity of fucoxanthin on Vero E6 cells using an MTT assay.
  • Figure 1b is an image of the plaque analysis results confirming the anti-Zika virus efficacy of fucoxanthin.
  • Figure 1c is a graph of the plaque analysis results confirming the anti-Zika virus efficacy of fucoxanthin.
  • Figure 1d is a graph showing the results of mRNA expression analysis of the NS1 protein, which confirmed the anti-Zika virus efficacy of fucoxanthin.
  • Figure 2a is a graph showing the results of the analysis of the expression of the inflammatory factor IFIT1 by Zika virus infection of fucoxanthin.
  • Figure 2b is a graph showing the results of the analysis of the expression of the inflammatory factor IFIT2 by Zika virus infection of fucoxanthin.
  • Figure 3a shows the results of analyzing the cytotoxicity of fucoxanthin on Vero cells using an MTT assay.
  • Figure 3b is a graph showing the analysis results confirming the SARS-CoV-2 infection inhibition efficacy of fucoxanthin.
  • the present invention provides a pharmaceutical composition for preventing or treating viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the above fucoxanthin may be a compound represented by the following chemical formula 1.
  • the 'Fucoxanthin' of the present invention is a substance mainly present in edible seaweeds such as rhubarb, Japanese tangle, kelp, and sea tangle, and is a carotenoid, one of the main pigments that gives these seaweeds a brown color, and is also called fucoxanthin.
  • This fucoxanthin is fat-soluble and is known to change from yellow to red as the number of conjugated double bonds increases, and since it has a very high degree of unsaturation, it is easily oxidized and destroyed by heat, light irradiation, oxygen, metal ions, peroxides, peroxidase, and chemicals, and has the characteristic of having a specific absorption band at a wavelength of 300 to 500 nm.
  • Fucoxanthin which has the above characteristics, is known to have physiological activities such as anticancer effects, anti-inflammatory effects, angiogenesis inhibition effects, anti-obesity effects, hyperlipidemia prevention effects, and fatty liver prevention effects.
  • fucoxanthin has excellent inhibitory activity against viruses, particularly coronavirus or Zika virus, and thus it is intended to apply the composition containing fucoxanthin as an active ingredient for preventing, improving or treating coronavirus or Zika virus infection.
  • the above coronavirus may be at least one selected from the group consisting of human coronavirus 229E (HCoV-229E), human coronavirus OC43 (HCoV-OC43), severe acute respiratory syndrome coronavirus (SARS-CoV), human coronavirus NL63 (HCoV-NL63, New Haven coronavirus), human coronavirus HKU1, Middle East respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2), and variants thereof.
  • HKU1 Middle East respiratory syndrome coronavirus
  • MERS-CoV Middle East respiratory syndrome coronavirus
  • SARS-Cov-2 severe acute respiratory syndrome coronavirus
  • the coronavirus infection may be selected from coronavirus disease-19 (COVID-19) caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome (SARS) caused by infection with severe acute respiratory syndrome coronavirus (SARS-CoV), and Middle East respiratory syndrome (MERS) caused by infection with Middle East respiratory syndrome coronavirus (MERS-CoV), and preferably coronavirus disease-19 (COVID-19) caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
  • the above SARS-Cov-2 variant may be at least one selected from the group consisting of alpha ( ⁇ ), beta ( ⁇ ), gamma ( ⁇ ), delta ( ⁇ ), and omicron ( ⁇ ) variants.
  • the fucoxanthin according to the present invention can be used in the form of a salt, preferably a pharmaceutically acceptable salt.
  • a salt an acid addition salt formed by a pharmaceutically acceptable free acid is preferable, and as the free acid, an organic acid and an inorganic acid can be used.
  • the organic acid includes, but is not limited to, citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid, and aspartic acid.
  • the inorganic acid includes, but is not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid.
  • the fucoxanthin according to the present invention may be commercially available, or may be isolated from nature or manufactured by a chemical synthesis method known in the art.
  • the separation can be carried out through the process.
  • the powder of the hat is extracted three times with 80% ethanol or methanol, concentrated under reduced pressure in a vacuum, suspended in an appropriate amount of distilled water, fractionated by adding an equal amount of chloroform, and then purified by fractionation using a silica column using chloroform and methanol as solvent systems, or a Sephadex LH-20 column using methanol as a solvent system.
  • the composition of the present invention is a pharmaceutical composition containing the above fucoxanthin as an active ingredient, and can be prepared using, in addition to the above active ingredient, a pharmaceutically suitable and physiologically acceptable excipient, and the excipient may be an excipient, a disintegrant, a sweetener, a binder, a coating agent, a swelling agent, a lubricant, a glidant, or a flavoring agent.
  • the pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions.
  • the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture, if desired or necessary.
  • Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweetener, acacia, tracheacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, etc.
  • acceptable pharmaceutical carriers are sterile and biocompatible, and include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary.
  • diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.
  • the present invention provides a use of a composition comprising fucoxanthin as an active ingredient for the manufacture of a medicament for the prevention or treatment of a viral infection.
  • the composition of the present invention comprising the above-described fucoxanthin as an active ingredient can be used particularly for the manufacture of a medicament for the prevention or treatment of a coronavirus or Zika virus infection.
  • the present invention provides a method for preventing or treating a viral infection, particularly a coronavirus or Zika virus infection, comprising a step of administering fucoxanthin or a pharmaceutically acceptable salt thereof to a subject in need thereof.
  • the term "subject” means a subject for which a disease is to be prevented, improved, or treated, and more specifically, may mean a mammal such as a human or non-human primate, mouse, dog, cat, horse, cow, etc., but is not limited thereto.
  • the concentration of fucoxanthin (or a pharmaceutically acceptable salt thereof) administered to the subject may be 0.1 to 100 uM, more preferably 10 to 50 uM, but is not limited thereto. Within the above concentration range, not only is the inhibitory effect of fucoxanthin against coronavirus or Zika virus excellent, but also no or very low cytotoxic effect against renal cells is observed.
  • prevention means any act of inhibiting the activity of a virus by administering the composition of the present invention, or thereby delaying the activity of a virus.
  • treatment means any action that inhibits the activity of a virus by administering the composition of the present invention, or thereby improves or beneficially alters a viral infection, and means an attempt to obtain a useful or desirable result, including a clinical outcome.
  • Useful or desirable clinical outcomes may include, but are not necessarily limited to, alleviation or improvement of one or more symptoms or conditions, reduction in the extent of the disease, stabilization of the disease state, inhibition of the occurrence of the disease, inhibition of the spread of the disease, delay or slowing of the progression of the disease, delay or slowing of the onset of the disease, improvement or alleviation of the disease state, and reduction (partial or complete).
  • treatment may mean prolonging the survival of a patient beyond what would be expected in the absence of treatment.
  • treatment may mean inhibiting the progression of a disease, temporarily slowing the progression of a disease, and more preferably relates to permanently stopping the progression of a disease. In the present invention, it may mean improving the survival of a patient by promoting the treatment of a viral infection, particularly an infection of a coronavirus or Zika virus.
  • coronavirus may mean improving patient survival by promoting the treatment of coronavirus disease-19 (COVID-19) caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
  • COVID-19 coronavirus disease-19
  • SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
  • the dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. The determination of the dosage based on these factors is within the level of those skilled in the art, and the dosage is generally in the range of 0.01 mg/kg/day to about 2000 mg/kg/day. A more preferred dosage is 0.1 mg/kg/day to 1000 mg/kg/day.
  • the administration may be administered once a day or divided into several times. The above dosage does not limit the scope of the present invention in any way.
  • the pharmaceutical composition of the present invention can be administered to mammals such as mice, livestock, and humans by various routes. All modes of administration can be envisaged, for example, oral, rectal, or intravenous, intramuscular, subcutaneous, intrauterine, epidural, or intracerebrovascular injection.
  • a pharmaceutical composition for preventing or treating a viral infection may additionally contain, in addition to an effective ingredient, any compound or natural extract known to have an activity whose safety has already been verified in order to increase and strengthen the effect of inhibiting the activity of the virus.
  • the present invention provides a food composition or health functional food composition for preventing or improving viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the above food or health functional food composition may additionally include an additive selected from the group consisting of flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, natural carbohydrates, nutrients, vitamins, thickeners, pH regulators, preservatives, and mixtures thereof.
  • the food composition of the present invention includes all forms such as functional food, nutritional supplement, health food, and food additives.
  • the food composition of the above type can be manufactured in various forms according to conventional methods known in the art.
  • the composition itself can be manufactured in the form of tea, juice, and drinks and consumed, or it can be granulated, encapsulated, and powdered and consumed.
  • the extract can be added to beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats and processed foods thereof (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.).
  • the composition of the present invention in the form of a food additive
  • the preferred content of fucoxanthin in the food composition of the present invention may be 0.001 to 50%, and preferably 0.01 to 30%, based on the total weight of the food composition.
  • the health functional food composition of the present invention can be manufactured in general dosage forms such as tablets, pills, granules, powders, liquids, hard capsules, soft capsules, etc., and can be manufactured in any form such as porridge, bread, beverages, bars, chocolate, cookies, tea, drinks, vitamin complexes, meat, sausages, candy, noodles, jelly, etc.
  • food-related acceptable carriers or additives such as the excipients described above may be used, and any carrier or additive known to be usable in the art for the manufacture of the formulation or form to be manufactured may be utilized.
  • the present invention provides a feed composition for preventing or improving viral infection, comprising fucoxanthin or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the feed composition may additionally contain a known feed supplement, food additive or feed additive, and may be manufactured in the form of fermented feed, compound feed, pellet form and silage.
  • Zika virus and SARS-CoV-2 were provided by the National Institute of Health, Korea Disease Control and Prevention Agency.
  • Vero E6 and Vero cells were purchased from ATCC.
  • DPBS, DMEM, EMEM/F12 powder, 100X L-glutamine, and FBS used in cell experiments were purchased from Gibco.
  • Penicillin/Streptomycin and Crystal violet were purchased from Sigma-Aldrich.
  • TRIzol and DEPC-water were purchased from Ambion, and Chloroform, Iso-protanol, and Ethanol were purchased from EMSURE.
  • High Capacity RNA to cDNA kit and Power SYBR Green PCR Master Mix were purchased from Applied Biosystems.
  • the primary antibody specific for anti-SARS-CoV-2 N protein was purchased from Sino Biological, and the secondary antibodies Alexa Fluor 488 goat anti-rabbit IgG and Hoechst 33342 were purchased from Molecular Probes.
  • Cytotoxicity was measured using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazoliumbromide (MTT) assay.
  • MTT 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazoliumbromide
  • 6-well plates were seeded with 1x106 Vero E6 cells per well and cultured until the cells became a monolayer. Afterwards, the cells were washed with DPBS and treated with samples by dispensing DMEM containing 2% FBS. Two hours after sample treatment, the cells were infected with Zika virus (0.01 MOI) and cultured at 37°C for 48 hours. Cells treated with DPBS instead of samples were used as non-infected cells (Mock).
  • Plaque assay was performed to determine the number of virus particles (Plaque-forming units, PFU) in each supernatant treated with virus and samples. Plaque assay was prepared by filling 6-well plates with a monolayer of cells, and serially diluting each supernatant 10-fold in DMEM. The cells were washed twice with DPBS, and a certain amount of the diluted supernatant was treated. The cells were gently shaken every 15 minutes to ensure that the virus was well absorbed by the cells, and the cells were cultured for 2 hours. After the supernatant was removed, 3 ml of DMEM/F12 with agarose was dispensed.
  • PFU virus-forming units
  • the 6-well plates were wrapped in foil, placed upside down in a CO2 incubator, and cultured for 4 days. After that, the cells were fixed for 1 hour by treating with 4% formaldehyde, and the agarose DMEM/F12 was removed along with the fixative. The cells were stained with 0.1% crystal violet solution, and the virus particles were counted. PFU per ml was calculated according to Equation 1 below.
  • PFU/ml Number of plaques / (Dilution factor ⁇ Volume of diluted virus/well)
  • RNA extraction was performed using the guanidium thiocyanate-phenol-chloroform extraction method. 1 ml of TRIzol was treated to cells treated with virus and samples, mixed well, and the mixed cells were collected in an e-tube. After that, 200 ⁇ l of chloroform was treated, vortexed for 15 seconds, left at room temperature for 3 minutes, and centrifuged at 12,000 rpm, 4°C for 15 minutes.
  • RNA obtained in this way was stored at -80°C and used in the experiment.
  • cDNA synthesis was performed using a High-Capacity RNA-to-cDNA kit.
  • RNA 2 ⁇ g was taken and mixed with 2X RT buffer mix, 20X RT enzyme mix, and DEPC-water in the correct ratio to make a total of 20 ⁇ l, and the reaction was performed at 37°C for 60 minutes and inactivated at 95°C for 5 minutes.
  • the cDNA was stored at -20°C and used in the experiment.
  • Real-time PCR was performed using a SYBR green-based detection method.
  • the synthesized cDNA was diluted to 1/40 and prepared, and SYBR green and target primer were mixed in the appropriate ratio.
  • Vero cells were seeded at 1.2 ⁇ 10 4 per well in 384-tissue culture plates. The following day, the cells were treated with compounds prepared in 10-point 2-fold serial dilutions in DMSO to a maximum concentration of 50 ⁇ M. Approximately 1 h after compound treatment, the cells were infected with SARS-CoV-2 (0.0125 MOI) in a BSL3 facility and cultured at 37°C for 24 h. The cells were then fixed with 4% paraformaldehyde (PFA) and permeabilized. The cells were then stained with anti-SARS-CoV-2 nucleocapsid (N) primary antibody, Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody, and Hoechst 33342. Fluorescence images of the infected cells were acquired using a high-throughput image analysis instrument, Operetta (Perkin Elmer).
  • the total cell number per well was calculated as the number of nuclei stained with Hoechst, and the number of infected cells was calculated as the number of cells expressing the viral N protein.
  • the infection ratio was calculated as the 'number of cells expressing N protein/total number of cells' in each well.
  • the infection inhibition rate (% Inhibition of infection) of each well was calculated by normalizing the average infection rates of the wells containing non-infected cells (mock) in the same plate and the average infection rates of the wells containing infected cells treated with 0.5% DMSO (v/v) to 100% and 0%, respectively.
  • the cell viability of the compound was expressed as '% Cell viability' in the graph by normalizing the cell number of each well to the average cell number of the mock group wells. Each value was obtained through two repeated experiments (technical duplicate).
  • the selectivity index (SI) value was calculated as CC 50 /IC 50 .
  • fucoxanthin The cytotoxicity of fucoxanthin against Vero E6 was analyzed using the MTT assay. As a result, fucoxanthin did not show a significant difference in cell viability compared to the control group at concentrations of 12.5, 25, and 50 ⁇ M, confirming that it had no cytotoxicity (see Fig. 1a).
  • the anti-Zika virus efficacy of fucoxanthin was evaluated by the inhibition rate of Zika virus plaque formation and the inhibition rate of Zika virus NS1 protein mRNA expression compared to the control group. As a result, it was confirmed that fucoxanthin significantly reduced the plaque formed upon Zika virus inoculation (see Fig. 1b).
  • the concentration of plaques formed in the control group treated only with Zika virus was found to be 15,000 ⁇ 566 PFU/ml
  • the concentrations of plaques formed in the experimental groups treated together with 12.5, 25, and 50 ⁇ M fucoxanthin were found to be 2,300 ⁇ 707, 400 ⁇ 283, and 300 ⁇ 424 PFU/ml, respectively, confirming that fucoxanthin inhibits the formation of plaques in a concentration-dependent manner (see Figure 1c).
  • the IFN-induced protein with tetratricopeptide repeats (IFIT) family is a group of proteins expressed in virus-infected cells, and is known to be an important element of the antiviral immune response by inhibiting the translation of viral proteins and viral replication. Therefore, the efficacy of fucoxanthin on the IFIT family expressed by Zika virus infection was confirmed. As a result, in cells infected with Zika virus, the expression of IFIT1 and IFIT2 significantly increased compared to the untreated Mock group.
  • Fucoxanthin was confirmed to not exhibit toxicity to Vero cells at concentrations of 0.1–50 ⁇ M (see Fig. 3a).
  • the anti-SARS-CoV-2 efficacy of fucoxanthin was evaluated by the infection inhibition rate based on immunofluorescence.
  • fucoxanthin exhibited infection inhibition rates of 1.02 ⁇ 0.54, 18.21 ⁇ 3.73, and 61.45 ⁇ 4.12% against SARS-CoV-2 at concentrations of 12.5, 25, and 50 ⁇ M, respectively, confirming that fucoxanthin exhibits antiviral efficacy (see Fig. 3b).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Biotechnology (AREA)
  • Polymers & Plastics (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Food Science & Technology (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Communicable Diseases (AREA)
  • Microbiology (AREA)
  • Medical Informatics (AREA)
  • Botany (AREA)
  • Nutrition Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Molecular Biology (AREA)
  • Animal Husbandry (AREA)
  • Zoology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne une composition pour prévenir, améliorer ou traiter des infections virales, la composition comprenant de la fucoxanthine. En particulier, la composition présente une excellente activité antivirale contre le coronavirus ou le virus Zika, et peut ainsi être efficacement appliquée à la prévention, à l'amélioration ou au traitement d'infections par le coronavirus ou le virus Zika.
PCT/KR2024/003350 2023-03-30 2024-03-18 Composition pharmaceutique pour la prévention ou le traitement d'infections virales, comprenant de la fucoxanthine Pending WO2024205087A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2023-0042165 2023-03-30
KR20230042165 2023-03-30
KR1020240034529A KR102694547B1 (ko) 2023-03-30 2024-03-12 푸코잔틴을 포함하는 바이러스 감염증의 예방 또는 치료용 약학적 조성물
KR10-2024-0034529 2024-03-12

Publications (1)

Publication Number Publication Date
WO2024205087A1 true WO2024205087A1 (fr) 2024-10-03

Family

ID=92376776

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2024/003350 Pending WO2024205087A1 (fr) 2023-03-30 2024-03-18 Composition pharmaceutique pour la prévention ou le traitement d'infections virales, comprenant de la fucoxanthine

Country Status (2)

Country Link
KR (1) KR102694547B1 (fr)
WO (1) WO2024205087A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180111342A (ko) * 2017-03-31 2018-10-11 조선대학교산학협력단 미역 추출물을 함유하는 코로나 바이러스의 예방 또는 치료용 조성물
KR20220109741A (ko) * 2021-01-29 2022-08-05 성균관대학교산학협력단 구멍쇠미역 추출물로부터 유래된 푸코이단을 유효성분으로 포함하는 신종코로나바이러스 예방 및 치료용 조성물
JP2022136956A (ja) * 2021-03-08 2022-09-21 株式会社 バイオミメティクスシンパシーズ アンジオテンシン転換酵素２（ａｃｅ２）及び／又はｔｍｐｒｓｓ２発現を阻害するための組成物

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20170125484A (ko) 2016-05-04 2017-11-15 서울대학교산학협력단 지카바이러스 감염 질환의 예방 또는 치료용 약제학적 조성물
KR102145197B1 (ko) 2020-03-10 2020-08-18 부광약품 주식회사 코로나바이러스를 치료하기 위한 l-뉴클레오사이드의 용도

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180111342A (ko) * 2017-03-31 2018-10-11 조선대학교산학협력단 미역 추출물을 함유하는 코로나 바이러스의 예방 또는 치료용 조성물
KR20220109741A (ko) * 2021-01-29 2022-08-05 성균관대학교산학협력단 구멍쇠미역 추출물로부터 유래된 푸코이단을 유효성분으로 포함하는 신종코로나바이러스 예방 및 치료용 조성물
JP2022136956A (ja) * 2021-03-08 2022-09-21 株式会社 バイオミメティクスシンパシーズ アンジオテンシン転換酵素２（ａｃｅ２）及び／又はｔｍｐｒｓｓ２発現を阻害するための組成物

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
TAMAMA KENICHI: "Potential benefits of dietary seaweeds as protection against COVID-19", NUTRITION REVIEWS, vol. 79, no. 7, 4 June 2021 (2021-06-04), pages 814 - 823, XP093216076, ISSN: 0029-6643, DOI: 10.1093/nutrit/nuaa126 *
YIM SUNG-KUN, KIM INHEE, WARREN BOYD, KIM JUNGWON, JUNG KYOOJIN, KU BOSUNG: "Antiviral Activity of Two Marine Carotenoids against SARS-CoV-2 Virus Entry In Silico and In Vitro", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL (MDPI), BASEL, CH, vol. 22, no. 12, Basel, CH , pages 6481, XP093216077, ISSN: 1422-0067, DOI: 10.3390/ijms22126481 *

Also Published As

Publication number Publication date
KR102694547B9 (ko) 2025-02-10
KR102694547B1 (ko) 2024-08-12

Similar Documents

Publication Publication Date Title
KR102204299B1 (ko) 담팔수 추출물을 유효성분으로 포함하는 코로나 바이러스 치료제
KR100998081B1 (ko) 라우릭산을 유효성분으로 함유하는 항바이러스제 조성물
WO2020231127A1 (fr) Souche présentant une activité améliorant la fonction hépatique, et son utilisation
KR20210116150A (ko) 테트란드린 또는 이의 유사체를 포함하는 코로나바이러스 감염 예방 또는 치료용 조성물
WO2020262799A1 (fr) Composition destinée à inhiber le virus de la grippe comprenant de la nodakénine et/ou de la nodakénétine
KR101708833B1 (ko) 커큐민을 포함하는 항바이러스 조성물
WO2024205087A1 (fr) Composition pharmaceutique pour la prévention ou le traitement d'infections virales, comprenant de la fucoxanthine
CN106038695B (zh) 鳄梨萃取物、鳄梨醇b及(2r,4r)-1,2,4-三羟基十七碳-16-炔的用途，以及包含鳄梨萃取物的保健食品
KR101653884B1 (ko) 항 콕사키바이러스 활성을 갖는 오리방풀 유래 화합물
CN102885841B (zh) 安石榴苷的应用
KR102570596B1 (ko) 지구자 추출물을 포함하는 코로나바이러스 감염 예방 또는 치료용 조성물
WO2015105294A1 (fr) Composition contenant du pentose et du xylitol utilisés comme principes actifs pour la suppression de la fixation de bactéries buccales
WO2016159584A2 (fr) Composition pour le renforcement de l'immunité innée et l'activité antivirale contenant un extrait de mori ramulus ou mori radicis cortex en tant que principe actif
Hurcan et al. A review of natural foods consumed during the COVID-19 pandemic life
RU2780346C1 (ru) Терапевтический агент против коронавируса, включающий экстракт elaeocarpus sylvestris
KR102694541B1 (ko) 플로로탄닌을 유효성분으로 포함하는 항바이러스제
KR102787715B1 (ko) 미세조류 추출물을 유효성분으로 포함하는 항바이러스제
KR102818485B1 (ko) 한약재 추출물을 유효성분으로 포함하는 코로나바이러스 감염증의 예방, 개선 또는 치료용 조성물
KR100792626B1 (ko) 카테킨 유도체 화합물을 유효성분으로 하는 독감 바이러스감염성 질환 예방 및 치료제
KR102748815B1 (ko) 돌외 추출물을 포함하는 코로나바이러스 감염 예방 또는 치료용 조성물
KR102856782B1 (ko) 산자나무 열매 추출물 또는 분획물을 포함하는 항바이러스용 약학적 조성물
WO2025258750A1 (fr) Agent antiviral comprenant un extrait d'ecklonia cava ou de phlorotanins en tant que principe actif
EP4151226A1 (fr) Agent thérapeutique contre le coronavirus comprenant un extrait de feuille de zanthoxylum piperitum en tant que principe actif
WO2018030857A1 (fr) Composition antivirale contenant un extrait de racine de panax notoginseng
WO2022065971A2 (fr) Composition pour la prévention ou le traitement d'une infection à coronavirus, comprenant un extrait de hovenia dulcis thunb

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24781078

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE