WO2013122575A2 - Small molecule having antiviral properties - Google Patents

Small molecule having antiviral properties Download PDF

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Publication number
WO2013122575A2
WO2013122575A2 PCT/US2012/025070 US2012025070W WO2013122575A2 WO 2013122575 A2 WO2013122575 A2 WO 2013122575A2 US 2012025070 W US2012025070 W US 2012025070W WO 2013122575 A2 WO2013122575 A2 WO 2013122575A2
Authority
WO
WIPO (PCT)
Prior art keywords
virus
small molecule
cells
monolayer
antiviral properties
Prior art date
Application number
PCT/US2012/025070
Other languages
English (en)
French (fr)
Other versions
WO2013122575A3 (en
Inventor
Oleg Nikolayevich CHUPAKHIN
Oleg Ivanovich KISILEV
Vladimir Leonidovich RUSINOV
Valery Nikolaevich CHARUSHIN
Nina Pablovna TUGOVA
Natliya Roseyvna MEDVEDERA
Evgeny Hartsisovich ULOMSKII
David Mark Evans
Ella Germanovna DYEVA
Original Assignee
Grl
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 Grl filed Critical Grl
Priority to EP12868920.5A priority Critical patent/EP2819517A2/en
Priority to JP2014557606A priority patent/JP2015507007A/ja
Priority to PCT/US2012/025070 priority patent/WO2013122575A2/en
Publication of WO2013122575A2 publication Critical patent/WO2013122575A2/en
Publication of WO2013122575A3 publication Critical patent/WO2013122575A3/en

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Classifications

    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • 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
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses

Definitions

  • the invention reiates to biologically active compounds possessing antiviral properties for the treatment and prevention of viral infections of animals and humans.
  • the invention can be used in hospitals, research laboratories, as well as livestock and poultry.
  • the closest in structure to the claimed compound from this series (1 ) can be regarded as a prototype.
  • the compound (1 ) in a concentration of 40 ug / mi in in vitro experiments the infectious titer of influenza ⁇ / ⁇ 3 ⁇ 2 and influenza virus
  • CC50 concentration at which 50% of the cells die
  • the invention is the small molecule (2) which has antiviral activity.
  • the small molecule (2) has demonstrated low toxicity.
  • Fig. 1 shows a scaffold for a small molecule.
  • Fig. 2 shows a small molecule based on the scaffold of Fig. 1 .
  • the compound produced from this synthesis is a pale yellow crystaliine solid, soluble in water, methanol, dimethyl sulfoxide, insoluble in benzene, ether and most other solvents.
  • the compound is orally ingestibie and show efficacy against viruses including the influenza virus.
  • Viruses Used a one-day monolayer culture of epithelial cells MDCK (dog kidney) Viruses. To assess the antiviral activity of the virus used the reference A / Puerto Rico/8/34, as well as pandemic influenza virus H1 1 v A/San kt-Peterburg/2/09 (similar to the so-called virus "swine fiu» A/Caiifornia/7/09).
  • the maximum tolerable concentrations of the compounds was determined by MTT test in ceil culture MDCK.
  • Testing of toxicity was carried out as follows: weighed weigh weighing 5 mg in a sterile test tube 5 ml. and the diluted growth medium for the cells MDCK (a-MEM, Bioiot, St. Russia) to a concentration of 1 mg / mL, thus obtaining a basic solution. More of the same medium made eight consecutive binary dilutions (500, 250, 125, 82.5, 31 .25, 1 5.13, 7.56 and 3.78 mg / mi, respectively), which was used for toxicity testing. The experience set in the four parallels for each concentration. One-day cell culture MDCK, grown in 98 ⁇ vveli plates (Costar), checked visually in an inverted microscope on the integrity of the monolayer.
  • the plates were washed twice with medium without serum, and then have made the test compound in appropriate concentrations in a volume of 100 ml in each well.
  • the plates were incubated for 72 h at 37 ° C in the presence of 5% C02, and then recorded the results of the experiment visually assessing the integrity of the monolayer compared with control cells, and by the MTT (quantitatively evaluating the viability of the ceils) using the tablet reader Hydex Chameleon.
  • Statistical analysis was performed using the program Statistica 6.0.
  • the plates were incubated for 60 min at 37 ° C in the presence of 5% CO 2, and then washed with medium to remove unbound viral particles to cells. Continue to make the drug in the wells with virus dilutions in 100 mi of the appropriate concentration. Each concentration of test compound was placed in four parallels for each virus dilution. Control wells filled with growth medium in the same volume. Also, the wells were left to re-test toxicity used concentrations.
  • the plates were incubated for 72 h at 37 0 G, and then recorded the results of the experiment visually assessing the integrity of the monolayer as compared to control ceils and the degree of cytopathic effect of virus in ceil culture, put hemagglutination reaction and MTT method was used to quantify cell viability using the tablet reader Hydex Chameleon.
  • Evaluation of toxicity, as well as evaluation of the antiviral action of drugs produced by three methods 1 ) monitoring and evaluation of the integrity of the monolayer cells under an inverted microscope. Implies a comparison of cell morphology of control wells with experienced and registration of changes under the influence of an agent (drug, virus, etc.). The change of morphology could include violating the integrity of the monolayer, changes in cell shape, expression of cytopathic effect in virus infection.
  • MTT The method used in the evaluation of drug sensitivity, based on the ability of dehydrogenases of living cells to restore the form of colorless 3- 4,5-dimethylthiazol-2-yl-2 ,5-difenilterarazola (MTT reagent) to blue crystalline farmazana soluble in DMSO or 96% alcohol. Saturation of color indicates the intensity of metabolic processes in cells, ie, the normal level of viability. The color intensity is recorded with a spectrophotometer and the absorbance values obtained can be used for statistical data processing.
  • the concentration of the drug during that kills 50% of a monolayer of cells in a test on the MDCK cells was 177 pg / ml.
  • Drug concentration at which all the cells survive the monolayer was 62.5 micrograms / ml, which was used to test the antiviral activity of the drug.
  • connection (2) in much smaller concentrations, in comparison with toxic, has a pronounced antiviral activity, reducing the titers of virus A / Puerto Rico/8/34 virus and pandemic influenza HlNlv A/Sankt- Peterburg/2/09 (A/California/7/09-podobny) for 3,0 lg concentrations 62.5 and 125 ug / ml, respectively, then there is a more active antiviral compound than with a prototype.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Molecular Biology (AREA)
  • Pulmonology (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
PCT/US2012/025070 2012-02-14 2012-02-14 Small molecule having antiviral properties WO2013122575A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP12868920.5A EP2819517A2 (en) 2012-02-14 2012-02-14 Small molecule having antiviral properties
JP2014557606A JP2015507007A (ja) 2012-02-14 2012-02-14 抗ウイルス性の特性を有する小分子
PCT/US2012/025070 WO2013122575A2 (en) 2012-02-14 2012-02-14 Small molecule having antiviral properties

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US2012/025070 WO2013122575A2 (en) 2012-02-14 2012-02-14 Small molecule having antiviral properties

Publications (2)

Publication Number Publication Date
WO2013122575A2 true WO2013122575A2 (en) 2013-08-22
WO2013122575A3 WO2013122575A3 (en) 2014-05-01

Family

ID=48984871

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2012/025070 WO2013122575A2 (en) 2012-02-14 2012-02-14 Small molecule having antiviral properties

Country Status (3)

Country Link
EP (1) EP2819517A2 (enrdf_load_stackoverflow)
JP (1) JP2015507007A (enrdf_load_stackoverflow)
WO (1) WO2013122575A2 (enrdf_load_stackoverflow)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015016818A1 (en) * 2013-07-29 2015-02-05 Ghias Asghar Novel azolo-1,2,4-triazine derived antiviral oral therapeutic
WO2018035509A3 (en) * 2016-08-19 2018-03-29 Grl, Inc. Small molecules having antiviral properties

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4405619A (en) * 1981-09-02 1983-09-20 Diamond Shamrock Corporation Antiinflammatory substituted-1,2,4-triazolo[4,3-b]-1,2,4-triazines
AR042052A1 (es) * 2002-11-15 2005-06-08 Vertex Pharma Diaminotriazoles utiles como inhibidores de proteinquinasas
RU2007123675A (ru) * 2004-11-24 2008-12-27 Новартис АГ (CH) Комбинации ингибиторов jak
RU2294936C1 (ru) * 2005-06-29 2007-03-10 Государственное Образовательное Учреждение Высшего Профессионального Образования Уральский Государственный Технический Университет-УПИ Натриевая соль 2-метилтио-6-нитро-1,2-4-триазоло[5,1-c]-1,2,4-триазин-7(4h)-она, дигидрат, обладающая противовирусной активностью
ATE484505T1 (de) * 2006-11-23 2010-10-15 Novartis Ag 5-sulfanylmethylä1,2,4ütriazolä1,5-aüpyrimidin- - olderivate als cxcr2-antagonisten
RU2343154C2 (ru) * 2007-01-31 2009-01-10 Общество с ограниченной ответственностью Научно-техническое предприятие "Лиганд" СПОСОБ ПОЛУЧЕНИЯ НАТРИЕВОЙ СОЛИ 2-МЕТИЛТИО-6-НИТРО-1,2,4-ТРИАЗОЛО[5,1-c]-1,2,4-ТРИАЗИН-7-ОНА, ДИГИДРАТА
RU2345080C2 (ru) * 2007-02-16 2009-01-27 ГОУ ВПО Уральский государственный технический университет-УПИ 4-(4'-ГИДРОКСИБУТИЛ)-6-ФЕНИЛ-1,2,4-ТРИАЗОЛО[5,1-c][1,2,4]ТРИАЗИН-7-ОН
RU2376307C1 (ru) * 2008-05-27 2009-12-20 Институт органического синтеза им. И.Я. Постовского Уральского отделения Российской академии наук 4-((Z)-4'-ГИДРОКСИБУТЕН-2'-ИЛ)-2-R-6-ФЕНИЛ-1,2,4-ТРИАЗОЛО[5,1-c][1,2,4]ТРИАЗИН-7-ОНЫ

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015016818A1 (en) * 2013-07-29 2015-02-05 Ghias Asghar Novel azolo-1,2,4-triazine derived antiviral oral therapeutic
WO2018035509A3 (en) * 2016-08-19 2018-03-29 Grl, Inc. Small molecules having antiviral properties

Also Published As

Publication number Publication date
EP2819517A2 (en) 2015-01-07
WO2013122575A3 (en) 2014-05-01
JP2015507007A (ja) 2015-03-05

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