Biomedical Chemistry: Research and Methods, 2018, 1(4), e00055

Application of the SPR Biosensor in Drug Prototypes Discovery with Human Cytochrome P450(51)
as an Example

L.A. Kaluzhskiy1,*, P.V. Ershov1, T.V. Shkel2, O.V. Gnedenko1, N.V. Ivanchina3, N.V. Strushkevich2, A.A. Kicha3, I.P. Grabovec2, A.A. Gilep2, S.A. Usanov2, V.A. Stonik3, A.S. Ivanov1

1Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia; *e-mail: la-kaluzhskiy@yandex.ru
2Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus,
5/2 Acad. Kuprevich Street, Minsk, 220141 Belarus
3Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of Russian Academy of Sciences,
159, 100 Let Vladivostoka av., Vladivostok, 690022 Russia

Key words: surface plasmon resonance (SPR); screening; human cytochrome P450(51); drug prototypes

DOI:10.18097/BMCRM00055

The whole version of this paper is available in Russian.

The development of the integral platform “From Gene to Lead”, consolidated computer methods, bioinformatics researches, and experimental approaches, significantly accelerated and optimized base structure search in the field of drug design. The necessity of the experimental verification of hundreds virtual structure hypothesis (results of molecular data base selections or de novo construction) requires demands the usage of the high-through out and sensitive methods for validation possible interaction between numerous of selected compounds and particular molecular targets and evaluation of affinity, kinetics and thermodynamics. Surface plasmon resonance (SPR) technology makes it possible to solve all these problems. In this article the methodical aspects of the optical SPR-biosensor usage in the field of drug prototypes selection are described using the human cytochrome P450(51) catalyzing one of the key step of cholesterol biosynthesis as an example.

Figure 1. Sensorgrams of CYP51A1 immobilization pH-scouting.
Figure 2. Typical sensorgram of CYP51A1 immobilization. The beginning of solution injections is marked with arrows.
Figure 3. Results of in vitro SPR screening of low-weight compound interactions
with CYP51A1 [4].
Figure 4. Results of in vitro SPR screening of low-weight compound interactions
with CYP51A1 [5].
Figure 5. Sensorgrams of levisculoside G interaction with CYP51A1.
Figure 6. Thermodynamic parameters of low-weight compounds interaction with CYP51A1.
1 – lanosterol, 2 – ketoconazole, 3 – holothurin A, 4 – asterosaponin P1, 5 – betulafolientriol,
6 – theasaponin, 7 – levisculoside G, 8 – henricioside Н1, 9 – digitonin; ΔG – change of Gibbs
free energy, ΔH – change of enthalpy, -TΔS – change of entropy

ACKNOWLEDGEMENTS

This study were performed within the framework of Program of Basic Scientific Research of Russian National Science Academies for 2013–2020 years (0518-2018- 0003) using the equipment of "Human Proteome" Core Facility of the Institute of Biomedical Chemistry which is supported by The Ministry of Education and Science of the Russian Federation under agreements №14.621.21.0017 (project ID RFMEFI62117X0017). Protein expression, spectrophotometric titration and enzyme assay were performed within the framework of the Belarusian State Program "High technologies and equipment" for 2016–2020 years (sub-program 1, “Innovative Biotechnologies – 2020”).

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