The use of Caco-2 line cells for predicting the xenobiotics biodavailability




Glycyrrhiza glabra root extract, glycyram, biopharmaceutical classification system, bioavailability


The biopermeability of glycyram, a biologically active substance from Glycyrrhiza glabra root was studied in the Problem Laboratory of Morphofunctional Studies of the National University of Pharmacy. Glycyram is a triterpene glycoside with a wide range of the pharmacological activity. However, the intensity of the activity depends on many pharmacokinetic characteristics of the drug, in particular on its bioavailability. Bioavailability in pharmacology reflects the ability of a medicinal substance to be absorbed in the body. It is generally assumed that passing drugs through the intestinal epithelium (a monolayer of cells) is a major barrier to the drug in its pathway into the circulatory system. The bioavailability of the glycyram substance was determined in vitro on a Caco-2 cell model reproducing most of the properties and characteristics of differentiated intestinal epithelial cells. The biopharmaceutical classification system (BCS) for active substances has been put into practice in the pharmaceutical sector of Ukraine. According to the existing BCS, the active substances are divided into 4 classes by their solubility in different media and permeability.

Aim. To determine which class of the biopharmaceutical classification system glycyram belongs to.

Materials and methods. The procedure for determining the permeability on a Caco-2 cell culture consisted of the following steps: cell cultivation, incubation of cells on a microporous filter (preparation of the test system), determination of the test suitability of the system (measurement of transepithelial electrical resistance, method calibration), determination of the permeability of the standardized substance glycyram compared to the internal standard – propranolol, quantitative determination of the test substance permeability by the HLC method with an UV or MS detector.

Results and discussion. During the experiment using the Caco-2 monolayer the average permeability value of glyceram (7.755 ± 0.517) – E-08 cm/sec was found. These data indicate that the glycyram substance has a low permeability through the epithelial layer of the intestine (less than 50 %). Considering the results of the previous studies showing a high solubility of glycyram and its low permeability it is possible to refer glycyram to class 3 according to the biopharmaceutical classification system.

Conclusions. The results of the studies obtained substantiate the feasibility of using glycyram-based dosage forms that would avoid passing the barrier of enterocytes of the small intestine, in particular pessaries, gels or creams.

Author Biographies

L. M. Maloshtan, National University of Pharmacy of the Ministry of Health of Ukraine

Doctor of Biology (Dr. habil), professor, head of the Department of Physiology and Human Anatomy

O. M. Shatalova, National University of Pharmacy of the Ministry of Health of Ukraine

Candidate of Medicine (Ph.D), associate professor of the Department of Physiology and Human Anatomy

O. A. Rukhmakova, National University of Pharmacy of the Ministry of Health of Ukraine

Doctor of Pharmacy (Dr. habil), associate professor of the Department of Technology of Drugs

L. O. Shakina, National University of Pharmacy of the Ministry of Health of Ukraine

Candidate of Biology (Ph.D), associate professor of the Department of Physiology and Human Anatomy


Ammosov, A. S., Litvinenko, V. I. (2002). Farmakom, 4, 1–8.

Rukhmakova, O. A., Yarnykh, T. G. (2014). Aktualni pytannia farmatsevtychnoi i medychnoi nauky ta praktyky, 1 (14), 47–49.

Shakina, L. O., Maloshtan, L. M. (2018). Proceeding from Mekhanizmy rozvytku patolohichnykh protsesiv i khvorob ta yikhnia farmakolohichna korektsiia: tezy dopovidei I nauk.-prakt. internet-konf. z mizhnar. uchastiu (18 zhovt. 2018 r.). (pp. 263–265). Kharkiv.

Yarnykh, T. G., Rukhmakova, O. A., Maloshtan, L. M., Yatsenko, O. Yu., Babenko, I. O. (2015). Ukraine. Patent na korysnu model № 95891 MPK A61K36/48, A61K9/06, A61R31/12. Biuleten, 1, 5.

Ipatova, O. M., Torhovskaia, T. I., Medvedeva, N. V., Prozorovskii, V. N. (2010). Biomeditcinskaia khimiia, 56 (1), 101–119. doi:

Holovenko, M. Ya., Baula, O. P., Borysiuk, I. Yu. (2010). Biofarmatsevtychna klasyfikatsiina systema. Kyiv, 300.

Maloshtan, L. M., Shatalova, O. M., Shakina, L. O. (2019). The Third International scientific congress of scientists of Europe: Proceedings of the III International Scientific Forum of Scientists «East–West» (January 11, 2019). Vienna.

Shokhin, I. E., Kulinich, Yu. I., Ramenskaia, H. V., Kukes, V. H. (2012). Biomeditsina, 3, 91. Available at:

Takenaka, T., Harada, N., Kuze, J., Chiba, M., Iwao, T., Matsunaga, T. (2016). Application of a Human Intestinal Epithelial Cell Monolayer to the Prediction of Oral Drug Absorption in Humans as a Superior Alternative to the Caco-2 Cell Monolayer. J Pharm Sci., 105 (2), 915-924. doi:

Bock, U., Flototto, T., Haltner, E. (2004). Validation of cell culture models for the intestine and the blood-brain barrier and comparison of drug permeation. ALTEX, 21 (3), 57–64.

Ramenskaia, H. V., Shokhin, I. E., Savchenko, A. Yu., Kulinich, Yu. I., Davidova, K. S. (2011). Biomeditsina, 2, 50.

Sambay, Y., De Angelis, I., Ranaldi, G. et al. (2005). The Caco-2 cell lina model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biology and Toxicology, 21, 1-26. doi:

Natoli, M., Leoni, B. D., D’Agnano, I., Zucco, F., Felsani, A. (2012). Good Caco-2 cell culture practices. Toxicology in Vitro, 26 (8), 1243-1246. doi:

Bondareva, I. al. (2008). Otcenka bioekvivalentnosti lekarstvennykh sredstv: metod. ukaz. Moscow: MZSR RF, 21.

Chumak, V. T. et al. (2007). Provedennia porivnialnykh doslidzhen in vitro dlia pidtverdzhennia ekvivalentnosti likarskykh zasobiv u tverdii dozovanii formi systemnoi dii : metod. rek. Kyiv: Morion, 30.

Hureieva, S. M., Albedkhani, O. S., Hroshovyi, T. A. (2015). Aktualni pytannia farmatsevtychnoi i medychnoi nauky ta praktyky, 3 (19), 38–43. doi:





Experimental and Clinical Pharmacology