4 research outputs found
Study on Hemostimulating Properties of Granulocyte-Macrophage Colony Stimulating Factor
The hemostimulating properties of granulocyte-macrophage colony-stimulating factor (GM-CSF) make possible its clinical use in alleviating side effects of anti-cancer radio- and chemotherapy, in bone marrow transplantation, and in the treatment of some primary immunodeficiency conditions associated with leukopenia. The State Research Center of Virology and Biotechnology βVectorβ of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing has developed a high-performance technology for production of recombinant human GM-CSF (rhGM-CSF) based on a recombinant E. coli strain. The aim of the study was to assess hemostimulating activity of the rhGM-CSF preparation obtained using the new developed technology, as observed in cell culture and in the mice model of myelosuppression induced by cyclophosphamide administration. Materials and methods: in vitro evaluation of rhGM-CSF hemostimulating activity was performed by MTT assay in the commercial HL-60 promyelocytic leukemia cell culture with preliminary suppression of cell growth rate by adding a low concentration of dimethyl sulfoxide to the medium. In vivo studies were carried out in CBA/CaLac mice with cyclophosphamide-induced myelosuppression. The hemostimulating properties of the drug were evaluated after subcutaneous administration of 1β175 Β΅g/kg doses for 4β5 days, following administration of a cytostatic agent. The total number of leukocytes and the content of their morphological forms were determined in blood samples taken at different time points after the drug administration. The statistical processing of the experimental data was based on analysis of variance using Statgraphics v. 5.0 software. Results: the proliferative activity of HL-60 cells incubated with the rhGM-CSF preparation for 72 hours was shown to be dose-dependent. The highest values of the increase in proliferative activity associated with an increase in the drug dose were observed in the concentration range from 0.04 to 0.64 ng/mL (proliferative activity increased by 11β18% when the dose was increased twofold). The experiments in mice demonstrated a two-phase pattern of the dose-dependent effect. The drug showed the highest hemostimulating effect at the dose of 90 Β΅g/kg. Conclusions: the rhGM-CSF preparation obtained using the new developed technology has a pronounced hemostimulating activity confirmed by both in vitro and in vivo test systems
ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ Π³Π΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠΈΡ ΡΠ²ΠΎΠΉΡΡΠ² ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ Π³ΡΠ°Π½ΡΠ»ΠΎΡΠΈΡΠ°ΡΠ½ΠΎ-ΠΌΠ°ΠΊΡΠΎΡΠ°Π³Π°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠΎΠ»ΠΎΠ½ΠΈΠ΅ΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅Π³ΠΎ ΡΠ°ΠΊΡΠΎΡΠ° ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°
The hemostimulating properties of granulocyte-macrophage colony-stimulating factor (GM-CSF) make possible its clinical use in alleviating side effects of anti-cancer radio- and chemotherapy, in bone marrow transplantation, and in the treatment of some primary immunodeficiency conditions associated with leukopenia. The State Research Center of Virology and Biotechnology βVectorβ of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing has developed a high-performance technology for production of recombinant human GM-CSF (rhGM-CSF) based on a recombinant E. coli strain. The aim of the study was to assess hemostimulating activity of the rhGM-CSF preparation obtained using the new developed technology, as observed in cell culture and in the mice model of myelosuppression induced by cyclophosphamide administration. Materials and methods: in vitro evaluation of rhGM-CSF hemostimulating activity was performed by MTT assay in the commercial HL-60 promyelocytic leukemia cell culture with preliminary suppression of cell growth rate by adding a low concentration of dimethyl sulfoxide to the medium. In vivo studies were carried out in CBA/CaLac mice with cyclophosphamide-induced myelosuppression. The hemostimulating properties of the drug were evaluated after subcutaneous administration of 1β175 Β΅g/kg doses for 4β5 days, following administration of a cytostatic agent. The total number of leukocytes and the content of their morphological forms were determined in blood samples taken at different time points after the drug administration. The statistical processing of the experimental data was based on analysis of variance using Statgraphics v. 5.0 software. Results: the proliferative activity of HL-60 cells incubated with the rhGM-CSF preparation for 72 hours was shown to be dose-dependent. The highest values of the increase in proliferative activity associated with an increase in the drug dose were observed in the concentration range from 0.04 to 0.64 ng/mL (proliferative activity increased by 11β18% when the dose was increased twofold). The experiments in mice demonstrated a two-phase pattern of the dose-dependent effect. The drug showed the highest hemostimulating effect at the dose of 90 Β΅g/kg. Conclusions: the rhGM-CSF preparation obtained using the new developed technology has a pronounced hemostimulating activity confirmed by both in vitro and in vivo test systems.Β ΠΠ΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠΈΠ΅ ΡΠ²ΠΎΠΉΡΡΠ²Π° Π³ΡΠ°Π½ΡΠ»ΠΎΡΠΈΡΠ°ΡΠ½ΠΎ-ΠΌΠ°ΠΊΡΠΎΡΠ°Π³Π°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠΎΠ»ΠΎΠ½ΠΈΠ΅ΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅Π³ΠΎ ΡΠ°ΠΊΡΠΎΡΠ° (ΠΠ-ΠΠ‘Π€) ΠΏΠΎΠ·Π²ΠΎΠ»ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°ΡΡ Π΅Π³ΠΎ Π² ΠΊΠ»ΠΈΠ½ΠΈΠΊΠ΅ Π΄Π»Ρ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΡ ΠΏΠΎΠ±ΠΎΡΠ½ΡΡ
ΡΡΡΠ΅ΠΊΡΠΎΠ² ΡΠ°Π΄ΠΈΠΎ- ΠΈ Ρ
ΠΈΠΌΠΈΠΎΡΠ΅ΡΠ°ΠΏΠΈΠΈ ΠΎΠ½ΠΊΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ
Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ, ΠΏΡΠΈ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΠΈ ΠΊΠΎΡΡΠ½ΠΎΠ³ΠΎ ΠΌΠΎΠ·Π³Π°, Π΄Π»Ρ Π»Π΅ΡΠ΅Π½ΠΈΡ Π½Π΅ΠΊΠΎΡΠΎΡΡΡ
ΠΏΠ΅ΡΠ²ΠΈΡΠ½ΡΡ
ΠΈΠΌΠΌΡΠ½ΠΎΠ΄Π΅ΡΠΈΡΠΈΡΠ½ΡΡ
ΡΠΎΡΡΠΎΡΠ½ΠΈΠΉ, ΡΠ²ΡΠ·Π°Π½Π½ΡΡ
Ρ Π»Π΅ΠΉΠΊΠΎΠΏΠ΅Π½ΠΈΠ΅ΠΉ. Π Π€ΠΠ£Π ΠΠΠ¦ ΠΠ Β«ΠΠ΅ΠΊΡΠΎΡΒ» Π ΠΎΡΠΏΠΎΡΡΠ΅Π±Π½Π°Π΄Π·ΠΎΡΠ° ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π° Π²ΡΡΠΎΠΊΠΎΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΠΈΡΠ΅Π»ΡΠ½Π°Ρ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΡ ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΡ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ ΠΠ-ΠΠ‘Π€ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° (ΡΡΠΠ-ΠΠ‘Π€) Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ ΡΡΠ°ΠΌΠΌΠ° E. coli. Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ: ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π³Π΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅ΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ ΠΠ-ΠΠ‘Π€ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°, ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΠΎΠ³ΠΎ ΠΏΠΎ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΠΎΠΉ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ, Π² ΠΊΡΠ»ΡΡΡΡΠ΅ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΈ Π½Π° ΠΌΠΎΠ΄Π΅Π»ΠΈ ΠΌΠΈΠ΅Π»ΠΎΡΡΠΏΡΠ΅ΡΡΠΈΠΈ Ρ ΠΌΡΡΠ΅ΠΉ, Π²ΡΠ·Π²Π°Π½Π½ΠΎΠΉ Π²Π²Π΅Π΄Π΅Π½ΠΈΠ΅ΠΌ ΡΠΈΠΊΠ»ΠΎΡΠΎΡΡΠ°Π½Π°. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: ΠΎΡΠ΅Π½ΠΊΡ Π³Π΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅ΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΡΡΠΠ-ΠΠ‘Π€ in vitro ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΠ’Π’-ΡΠ΅ΡΡΠ° Π½Π° ΠΊΠΎΠΌΠΌΠ΅ΡΡΠ΅ΡΠΊΠΎΠΉ ΠΊΡΠ»ΡΡΡΡΠ΅ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΡΠΎΠΌΠΈΠ΅Π»ΠΎΡΠΈΡΠ°ΡΠ½ΠΎΠΉ Π»Π΅ΠΉΠΊΠ΅ΠΌΠΈΠΈ HL-60, ΡΠΊΠΎΡΠΎΡΡΡ ΡΠΎΡΡΠ° ΠΊΠΎΡΠΎΡΡΡ
ΠΏΡΠ΅Π΄Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΏΠΎΠ΄Π°Π²Π»ΡΠ»ΠΈ Π΄ΠΎΠ±Π°Π²Π»Π΅Π½ΠΈΠ΅ΠΌ Π² ΡΡΠ΅Π΄Ρ Π½ΠΈΠ·ΠΊΠΎΠΉ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π΄ΠΈΠΌΠ΅ΡΠΈΠ»ΡΡΠ»ΡΡΠΎΠΊΡΠΈΠ΄Π°. ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ in vivo ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Π½Π° ΠΌΡΡΠ°Ρ
Π»ΠΈΠ½ΠΈΠΈ Π‘ΠΠ/CalaΡ Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
ΠΌΠΈΠ΅Π»ΠΎΡΡΠΏΡΠ΅ΡΡΠΈΠΈ, Π²ΡΠ·Π²Π°Π½Π½ΠΎΠΉ Π²Π²Π΅Π΄Π΅Π½ΠΈΠ΅ΠΌ ΡΠΈΠΊΠ»ΠΎΡΠΎΡΡΠ°Π½Π°. ΠΠ΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠΈΠ΅ ΡΠ²ΠΎΠΉΡΡΠ²Π° ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ ΠΏΡΠΈ Π΅Π³ΠΎ ΠΏΠΎΠ΄ΠΊΠΎΠΆΠ½ΠΎΠΌ Π²Π²Π΅Π΄Π΅Π½ΠΈΠΈ Π² Π΄ΠΈΠ°ΠΏΠ°Π·ΠΎΠ½Π΅ Π΄ΠΎΠ· ΠΎΡ 1 Π΄ΠΎ 175 ΠΌΠΊΠ³/ΠΊΠ³ Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 4β5 ΡΡΡΠΎΠΊ ΠΏΠΎΡΠ»Π΅ Π²Π²Π΅Π΄Π΅Π½ΠΈΡ ΡΠΈΡΠΎΡΡΠ°ΡΠΈΠΊΠ°. Π ΠΎΠ±ΡΠ°Π·ΡΠ°Ρ
ΠΊΡΠΎΠ²ΠΈ, Π²Π·ΡΡΡΡ
Π² ΡΠ°Π·Π½ΡΠ΅ ΡΡΠΎΠΊΠΈ ΠΏΠΎΡΠ»Π΅ Π²Π²Π΅Π΄Π΅Π½ΠΈΡ, ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΠΎΠ±ΡΠ΅Π΅ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π»Π΅ΠΉΠΊΠΎΡΠΈΡΠΎΠ² ΠΈ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ ΠΈΡ
ΠΌΠΎΡΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΠΎΡΠΌ. ΠΠ°Π½Π½ΡΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ° ΠΎΠ±ΡΠ°Π±Π°ΡΡΠ²Π°Π»ΠΈ ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ Π²Π°ΡΠΈΠ°ΡΠΈΠΎΠ½Π½ΠΎΠΉ ΡΡΠ°ΡΠΈΡΡΠΈΠΊΠΈ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΏΠ°ΠΊΠ΅ΡΠ° ΠΏΡΠΎΠ³ΡΠ°ΠΌΠΌ Statgraphics, v. 5.0. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½Π°Ρ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ HL-60, ΠΈΠ½ΠΊΡΠ±ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Ρ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠΌ ΡΡΠΠ-ΠΠ‘Π€ Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 72 Ρ, Π½ΠΎΡΠΈΠ»Π° Π΄ΠΎΠ·ΠΎΠ·Π°Π²ΠΈΡΠΈΠΌΡΠΉ Ρ
Π°ΡΠ°ΠΊΡΠ΅Ρ. ΠΠ°ΠΈΠ±ΠΎΠ»Π΅Π΅ Π²ΡΡΠΎΠΊΠΈΠ΅ Π·Π½Π°ΡΠ΅Π½ΠΈΡ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΏΡΠΈ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠΈ Π΄ΠΎΠ·Ρ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° Π½Π°Π±Π»ΡΠ΄Π°Π»ΠΈΡΡ Π² Π΄ΠΈΠ°ΠΏΠ°Π·ΠΎΠ½Π΅ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΉ ΠΎΡ 0,04 Π΄ΠΎ 0,64 Π½Π³/ΠΌΠ» (ΠΏΡΠΈΡΠΎΡΡ Π·Π½Π°ΡΠ΅Π½ΠΈΠΉ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Ρ ΠΏΡΠΈ Π΄Π²ΡΠΊΡΠ°ΡΠ½ΠΎΠΌ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠΈ Π΄ΠΎΠ·Ρ ΡΠΎΡΡΠ°Π²Π»ΡΠ» 11β18%). Π ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Ρ
Π½Π° ΠΌΡΡΠ°Ρ
ΠΏΡΠΎΠ΄Π΅ΠΌΠΎΠ½ΡΡΡΠΈΡΠΎΠ²Π°Π½ Π΄Π²ΡΡ
ΡΠ°Π·Π½ΡΠΉ Ρ
Π°ΡΠ°ΠΊΡΠ΅Ρ Π΄ΠΎΠ·ΠΎΠ²ΠΎΠΉ Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΡΡΡΠ΅ΠΊΡΠ°. ΠΠ°ΠΈΠ±ΠΎΠ»ΡΡΡΡ Π³Π΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΏΡΠ΅ΠΏΠ°ΡΠ°Ρ ΠΏΡΠΎΡΠ²Π»ΡΠ» Π² Π΄ΠΎΠ·Π΅ 90 ΠΌΠΊΠ³/ΠΊΠ³. ΠΡΠ²ΠΎΠ΄Ρ: ΠΏΡΠ΅ΠΏΠ°ΡΠ°Ρ ΡΡΠΠ-ΠΠ‘Π€, ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΠΉ ΠΏΠΎ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΠΎΠΉ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ, ΠΎΠ±Π»Π°Π΄Π°Π΅Ρ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΠΎΠΉ Π³Π΅ΠΌΠΎΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅ΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡΡ, ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½Π½ΠΎΠΉ Π² ΡΠΈΡΡΠ΅ΠΌΠ°Ρ
in vitro ΠΈ in vivo
Safety and Pharmacokinetics of the Substance of the Anti-Smallpox Drug NIOCH-14 after Oral Administration to Laboratory Animals
Background: Since most of the modern human population has no anti-smallpox immunity, it is extremely important to develop and implement effective drugs for the treatment of smallpox and other orthopoxvirus infections. The objective of this study is to determine the main characteristics of the chemical substance NIOCH-14 and its safety and bioavailability in the body of laboratory animals. Methods: The safety of NIOCH-14 upon single- or multiple-dose intragastric administration was assessed according to its effect on the main hematological and pathomorphological parameters of laboratory mice and rats. In order to evaluate the pharmacokinetic parameters of NIOCH-14 administered orally, a concentration of ST-246, the active metabolite of NIOCH-14, in mouse blood and organs was determined by tandem mass spectrometry and liquid chromatography. Results: The intragastric administration of NIOCH-14 at a dose of 5 g/kg body weight caused neither death nor signs of intoxication in mice. The intragastric administration of NIOCH-14 to mice and rats at doses of 50 and 150 Β΅g/g body weight either as a single dose or once daily during 30 days did not cause animal death or critical changes in hematological parameters and the microstructure of internal organs. The tissue availability of NIOCH-14 administered orally to the mice at a dose of 50 Β΅g/g body weight, which was calculated according to concentrations of its active metabolite ST-246 for the lungs, liver, kidney, brain, and spleen, was 100, 69.6, 63.3, 26.8 and 20.3%, respectively. The absolute bioavailability of the NIOCH-14 administered orally to mice at a dose of 50 Β΅g/g body weight was 22.8%. Conclusion: Along with the previously determined efficacy against orthopoxviruses, including the smallpox virus, the substance NIOCH-14 was shown to be safe and bioavailable in laboratory animal experiments