Особенности фармакокинетики 7-о-гентиобиозида формононетина определяют его гемостатическую активность у крыс

Aims: 1. to study the comparative effect of 7-O-gentiobioside formononetin (GBF) from the roots of M. amurensis on platelet and coagulative hemostasis under prolonged enteral and intraperitoneal administration to rats; 2. to determine the metabol- ic products of GBF in blood and urine of rats under enteral and intraperitoneal administration. Materials and methods. The study was performed on male Wistar rats weighing 220–280 grams. In the first series of exper- iments animals received GBF enterally and intraperitoneally at a dose of 25 mg/kg for 10 days. At the end of the administra- tion period we determined ADP-stimulated platelets aggregation, values of activated partial thromboplastin time, prothrom- bin time, thrombin time, fibrinogen level, content of soluble fibrin-monomer complexes, antithrombin III activity. Thromboelas- tometry was used for integral hemostasis analysis. In the second series of experiments GBF was administered enterally to 6 animals (group 1) in a dose of 150 mg/kg. Blood was collected 2 and 4 hours after GBF administration, urine was collected 8 hours after GBF administration. In 6 animals (group 2) the same manipulations were performed after intraperitoneal GBF ad- ministration in the same dose. Special samples were prepared for chromatographic-mass spectrometric analysis; the obtained fractions were analyzed on Shimadzu HPLC-PDA-MS chromatomass spectrometer. The mass spectra were recorded using the electrospray ionization method in the positive and negative ion mode (1.50 kV) in the m/z range 100–800. The obtained results were processed by a statistical method of variational series using the Mann-Whitney test. Results. Enteral administration of isoflavonoid GBF had a significant effect on platelet and coagulative hemostasis in rats; so getting the gastrointestinal tract GBF inhibited platelet aggregation and blood coagulation. Almost 10-fold decreasing in ADP-aggregation was registered in comparison with control animals. Enteral GBF administration was followed by a distinct and deep shift of the main chronometric recording parameters; under intraperitoneal GBF administration its suppressive ac- tion on hemostasis was not observed. The main stages of GBF metabolism under its enteral administration in rats were de- termined. The products of GBF biotransformation in rat’s intestine were revealed. Conclusions. 1. For the first time in experiments on rats the ability of isoflavonoid GBF isolated from the roots of M. amuren- sis was found to inhibit some parameters of platelet and coagulative hemostasis under enteral but not intraperitoneal admin- istration. This fact is of great practical importance because it opens the prospect for creating of new oral drugs that can re- duce the risk of thrombosis in various cardiovascular diseases. 2. We determined the main stages of GBF metabolism in rats that affected its bioavailability under enteral administration. 3. The biological activity of the studied isoflavonoid in rats is due to its biotransformation into formononetin, daidzein, and other metabolic products formed under the influence of the intesti- nal microbiota of animals. REFERENCES Plotnikov T.M., Anishchenko A.M., Plotnikov M.B. Phytoestro- gens: mechanisms of correction of cardiovascular complications [Fitoestrogeny: mekhanizmy korrekcii serdechno-sosudistyh oslozhnenij]. Eksperimental’naya i klinicheskaya farmakologiya. 2017; 80 (1): 39–44 (in Russ.). Anishcenko A.M. Hemoreological effects of complex isoflavo- noid preparation in ovariectomized rats. Bull Exp Biol Med. 2013; 154 (6): 755–7. Jacques P.F., Cassidy A., Peterson J.J., Dwyer J.T. Dietary flavo- noid intakes and CVD incidence in the Framingham Offspring Cohort. Br J Nutr. 2015; 114 (9): 1496–503. Kulesh N.I., Fedoreev S.A., Veselova M.V. et al. Influence of isofla- vonoids from the roots of Maackia amurensis on metabolic reac- tions of the liver in experimental toxic hepatitis [Vliyanie izoflavonoidov iz kornej Maackia amurensis na metabolicheskie reakcii pecheni pri eksperimental’nom toksicheskom gepatite]. Himiko- farmacevticheskij zhurnal. 2016; 50 (7): 21–7 (in Russ.). Kulesh N.I., Fedoreyev S.A., Veselova M.V. et al. Antioxidant activ- ity of the isoflavonoids from the roots of Maackia amurensis. Nat Prod Comm. 2013; 8 (5): 589–92. Thilakarathna S.H., Rupasinghe H.P.V. Flavonoid bioavailabili- ty and attempts for bioavailability enhancement. Nutrients. 2013; 5: 3367–87. Marin L., Miguelez E.M., Villar C.J., Lombo F. Bioavailability of dietary polyphenols and gut microbiota metabolism: Anti- microbial properties. BioMed Res Int. 2015; 2015: 905215. DOI: 10.1155.2015/905215. Makarova M.N. Bioavailability and metabolism of flavonoids [Bio- dostupnost’ i metabolizm flavonoidov]. Voprosy pitaniya. 2011; 80 (3): 4–12 (in Russ.). Kroon P.A., Clifford M.N., Crozier A. et al. How should we assess the effects of exposure to dietary polyphenols in vitro? Am J Clin Nutr. 2004; 8: 15–21. Shelnutt S.R., Cimino C.O., Wiggins P.A. et al. Pharmacokinetics of the glucuronide and sulfate conjugates of genistein and daidze- in in men and women after consumption of a soy beverage. Am J Clin Nutr. 2002; 76: 588–94. Zamyatin S.V., Zverev Ya.F., Momot A.P. et al. Effect of 7-O-genti- bioside formononetin on platelet hemostasis in rats [Vliyanie 7-O-gentiobiozida formononetina na pokazateli trombocitar- nogo gemostaza u krys]. Tromboz, gemostaz i reologiya. 2016; 2: 55–8 (in Russ.). Zamyatin S.V., Zverev Ya.F., Momot A.P. et al. Effect of 7-O-genti- bioside formononetin on coagulation hemostasis and fibrinolysis in rats [Vliyanie 7-O-gentiobiozida formononetina na pokazateli koagulyacionnogo gemostaza i fibrinoliz u krys]. Tromboz, gemo- staz i reologiya. 2016; 3: 73–6 (in Russ.). Momot A.P. Pathology of hemostasis. Principles and algorithms of clinical and laboratory diagnostics [Patologiya gemostaza. Principy i algoritmy kliniko-laboratornoj diagnostiki]. SPb.: FormaT. 2006: 208 s (in Russ.). Avdushkina L.A., Vavilova T.V., Zybina N.N. Thromboelastogra- phy/thromboelastometry in assessing hemostasis: past and present. Reference limits [Metod tromboelastogra-fii/tromboelastometrii v ocenke sistemy gemostaza: proshloe i nastoyashchee. Referentnye predely]. Kliniko-laboratornyj konsilium. 2009; 5: 26–33 (in Russ.). Barkagan Z.S., Momot A.P. Diagnosis and controlled therapy of haemostasis disorders [Diagnostika i kontroliruemaya terapiya narushenij gemostaza]. Moskva: N’yudiamed. 2008: 292 s (in Russ.). Dolgov V.V., Svirin P.N. Laboratory diagnostics of hemostasis dis- orders [Laboratornaya diagnostika narushenij gemostaza]. Mosk- va-Tver’: Triada. 2005: 227 s (in Russ.). Fuentes E., Palomo I. Antiplatelet effects of natural bioactive com- pounds by multiple targets: Food and drug interactions. J Funct Foods. 2014; 6: 73–81. Karlickova J., Riha M., Filipsky T. et al. Antiplatelet effects of flavo- noids mediated by inhibition of arachidonic acid based pathway. Published online. Planta med © Georg Thieme Verlag KG, Stutt- gart, New York. 2015. DOI: 10.1055/s-0035–1557902. Navarro-Nuсez L., Lozano M.L., Palomo M. et al. Apigenin inhib- its platelet adhesion and thrombus formation and synergizes with aspirin in the suppression of the arachidonic acid pathway. J Agric Food Chem. 2008; 56 (9): 2970–6. Navarro-Nuсez L., CastilloJ., Lozano M.L. et al. Thromboxane A2 receptor antagonism by flavonoids: structure-activity relationships. J Agric Food Chem. 2009; 57 (4): 1589–94. Correia-da-Silva M., Susa E., Duarte B. et al. Flavonoids with an oligopolysulfated moiety: a new class of anticoagulant agents. J Med Chem. 2011; 54 (1): 95–106. Bijak M., Ponczek M.B., Nowak P. Polyphenol compounds belong- ing to flavonoids inhibit activity of coagulation factor X. J Biol Mac- romol. 2014; 65: 129–35.