Кафедра "Біотехнологія, біофізика та аналітична хімія"

Постійне посилання колекціїhttps://repository.kpi.kharkov.ua/handle/KhPI-Press/15

Офіційний сайт кафедри http://web.kpi.kharkov.ua/biotech

Кафедра "Бiотехнологiя, біофізика та аналiтична хiмiя" була створена у 1998 році на базі кафедри "Аналітична хімія", яка у 1940 році була виділена з кафедри хімії в самостійну кафедру. Ініціатива створення кафедри належить доктору технічних наук, професору Миколі Федосовичу Клещеву.

Кафедра входить до складу Навчально-наукового інституту хімічних технологій та інженерії Національного технічного університету "Харківський політехнічний інститут". Кафедра провадить освітню, методичну та наукову діяльність у галузі знань "Хімічна та біоінженерія". Крім теоретичних основ біотехнології, велику увагу було приділяється контролю якості і сертифікації біотехнологічної продукції.

У складі науково-педагогічного колективу кафедри працюють: 2 доктора наук: 1 – технічних, 1 – фармацевтичних; 8 кандидатів наук: 3 – біологічних, 5 – технічних; 2 співробітника мають звання професора, 6 – доцента.

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Preparation and in-vivo evaluation of cytochrome-C-containing liposomes
(GOVI Verlag, Germany, 2017) Katsai, O. G.; Ruban, O. A.; Krasnopolsky, Yu. M.
This study investigates the development of a method for obtaining cytochrome C-containing liposomes (LS-Cyt), and evaluates their stability and specific activity. LS-Cyt were intended for the therapy of ophthalmic diseases. LS-Cyt were prepared by high pressure homogenization technique and lyophilized to obtain freeze-dried LS-Cyt. It was proposed to use anionic phospholipid- dipalmitoylphosphatidylglycerol (DPPG-Na) and phosphatidylcholine (PC) in a nanoparticulate composition. Were investigated various concentrations of lactose and trehalose as cryoprotectants. Samples with a lactose concentration of 6% showed the best results in terms of the emulsion formation time, encapsulation and preservation of nanosize. The main technological parameters for the obtained freeze-dried LS-Cyt were encapsulation of no less than 95% of cytochrome C (Cyt C), particle size of 140-170 nm, pH of 6.85±0.1, osmolarity of 330±3 mOsmol/kg, a lysophosphatidylcholine content (LPC) of 0.65±0.05 % of the total of lipids. Stability of the freeze-dried LS-Cyt during storage was established. The freeze-dried LS-Cyt was kept for 1 year in a light protected place at the temperature of -15 °C. No changes in the composition of LS-Cyt samples were detected over the observation period. Preclinical in-vivo research was conducted, namely the evaluation of specific activity on the model of the penetrating corneal injury. It was established that use of LS-Cyt contributes to a more rapid process of tissue regeneration and reduction of the inflammatory response in comparison with a non-liposomal dosage form.
Experimental study of liposomal docetaxel analysis of docetaxel incorporation and stability
(Morion LLC, 2017) Krasnopolsky, Yu. M.; Dudnichenko, А. S.
The article presents the results of developing the composition and technology of obtaining the liposomal form of docetaxel. The effect of the phospholipid composition of the membrane, ionic strength, pH, temperature, cryoprotectant type, and other factors on the stability of liposomes and the docetaxel incorporation has been considered. Results: Reduction of toxicity of the liposomal form of docetaxel (LD50 – 137 ± 7.7 mg/kg) was found in comparison with its free form (LD50 – 101 ± 6.3 mg/kg). Preservation of nanosize particle after lyophilization has been shown. Conclusions: As a result of the studies, the optimal composition and technological scheme for obtaining liposomes containing docetaxel have been developed allowing large-scale production of docetaxel in liposomal form.
Study of antioxidant activity of liposomal forms of quercetin and curcumin in ischemic heart disease
(Termedia Publishing House, Poland, 2020) Pylypenko, D. M.; Gorbach, T. V.; Krasnopolsky, Yu. M.
Quercetin and curcumin are plant polyphenolic antioxidants with proven pharmacological efficacy. Their use is, however, limited due to low bioavailability and oral administration route. The encapsulation of lipophilic compounds in liposomes enables to increase their bioavailability and to create an injectable form. The present study aimed to comparatively investigate the antioxidant activity of a complex liposomal preparation containing two lipophilic antioxidants (quercetin and curcumin) and their monopreparations in liposomal forms. This study was conducted on Wistar line rats with experimental model of ischemic heart disease. Oxidative stress markers such as total antioxidant activity, malondialdehyde, and peroxidized proteins were analyzed in blood serum and cardiac tissue. Ischemic heart disease is accompanied with lipid peroxidation and changes in the activity of the antioxidant system. The obtained results demonstrated the antioxidant activity of monopreparations of curcumin and quercetin and their complex in liposomal forms. Quercetin and curcumin showed different antioxidant activities in terms of oxidative stress markers. The complex of the two antioxidants showed the synergistic effect of their lipophilic compounds in liposomal forms, which led to the normalization of test parameters according to the level of the control sample.
"Quality by design" in liposomal drugs creation
(Національна академія наук України, 2020) Krasnopolsky, Yu. M.; Pylypenko, D. M.
Nanobiotechnological preparations creation is one of the promising areas of modern pharmacy, since it allows creating products of a qualitatively new level. The procedure development, based on an understanding of the product characteristics and the technological process, confirmed by reliable scientific data. The article is devoted to the pharmaceutical development of liposomal drugs. On the basis of our own experience in the development of liposomal medicinal forms, as well as on the basis of literature data, the main components in their composition were detected and these components impact on the quality indicators of liposomes were studied. Individual lipids function in nanoparticle membrane and their interaction, which determines the stability both in the technological process and upon storage of the product, were considered. The advantages and disadvantages of cholesterol incorporation into liposomes with hydrophilic and hydrophobic active pharmaceutical ingredients were described. Cryoprotectors and buffer systems role in ensuring nanopreparation stability is discussed.
The influence of complex liposomal antioxidant preparations on biological oxidative stress markers in ischemic heart disease
(Publishing center ESCBM, Česká republika, 2020) Pylypenko, D. M.; Gorbach, T. V.; Krasnopolsky, Yu. M.
Nanobiotechnological obtaining of liposomal forms of antioxidant preparations based on bioflavonoids
(НВП ПП "Технологічний Центр", 2019) Pylypenko, D. M.; Prokhorov, Vitaliy; Dudnichenko, Olexander; Krasnopolsky, Yu. M.
Більшість патологічних станів супроводжується перекисним окисненням ліпідів і накопиченням продуктів оксидативного стресу. Відома антиоксидантна дія природних гідрофобних сполук, таких як кверцетин, убіхінон, куркумін, вітамін Е та ін. Крім того відомо, що ці біологічно активні сполуки діють на різні ланки антиоксидантної системи. Однак, їх використання у складі парентеральних препаратів ускладнено, враховуючи їх гідрофобність. Для підвищення біодоступності ліпофільних антиоксидантів і створення їх водорозчинної форми використовують наночастинки, наприклад, ліпосоми. Метою роботи є розробка ліпосомального препарату з соінкапсуляцією двох гідрофобних антиоксидантів, а саме куркуміну та кверцетину. Методи. При розробці використовувалися технологічні методи отримання ліпосом та аналітичні фізико-хімічні, хроматографічні (ВЕРХ, ТШХ, ГРХ), методи визначення розміру часток, рН. Результати. В результаті проведеного дослідження запропоновано склад і технологію одержання ліпосомальної форми куркуміну та його композиції з кверцетином. Вивчено вплив жирно-кислотного складу ліпідів, співвідношення «ліпід: діюча речовина» та технологічних умов на утворення ліпосом та ступінь інкапсуляції активного фармацевтичного інгредієнта. Вивчено залежність розмірів наночастинок від значення тиску і кількості циклів гомогенізації. Отримано ліофілізований продукт зі ступенем включення гідрофобних антиоксидантів не менше 85 %. Проведено вивчення фізико-хімічних властивостей отриманих зразків. Висновки. Запропоновано технологічну схему одержання комплексного препарату, що містить куркумін і кверцетин, що включає отримання ліпідної плівки, гідратацію компонентів, гомогенізацію високого тиску, стерилізуючу фільтрацію і ліофілізацію.
A Study of Oxidative Stress Markers when Using the Liposomal Antioxidant Complex
(Hellenic Pharmaceutical Society, 2019) Pylypenko, D. M.; Gorbach, T. V.; Katsai, O. G.; Grigoryeva, A. S.; Krasnopolsky, Yu. M.
The aim of the study was to create the complex liposomal preparation containing Coenzyme Q10 and Quercetin and learn its antioxidant activity. The liposomes were obtained the lipid film method followed by emulsification with PBS, extrusion and sterilizing filtration. The emulsion was lyophilized with lactose as a cryoprotectant. The ratio of Coenzyme Q10 and Quercetin in complex liposomal composition was 1:1. The particles size and the degree of inclusion were determined. Pharmacological study was carried out on Wistar line rats with experimental ischemic heart disease model. The samples of complex liposomal composition and monocompositions of Coenzyme Q10 and Quercetin were intravenously administered daily for 5 days at a dose of 10 mg/kg. Oxidative stress markers (levels of malondialdehyde, conjugated dienes, sulfhydryl groups, protein peroxidation, enzyme superoxide dismutase catalase and total antioxidant activity) were analyzed in blood serum and cardiac tissue of the rats. The obtained complex composition was characterized by a particle size of up to 220 nm and at least 90.0 % of the included Coenzyme Q10 and Quercetin. A decrease in the level of lipid oxidation products and an increase in the activity of antioxidant system using the complex liposomal preparation containing Coenzyme Q10 and Quercetin compared to its monocompositions in liposomal form were shown.
"Quality-by-Design" approach to the development of a dosage form for the liposomal delivery system of cytochrome С
(Hellenic Pharmaceutical Society, 2018) Katsai, O. G.; Ruban, O. A.; Krasnopolsky, Yu. M.
Cytochrome C (Cyt C) is a natural catalyst for cellular respiration. Aqueous solutions of Cyt C are used in cardiology, neurology and ophthalmology, where they manifests a high activity towards free radicals and bind aggressive molecules of oxidants. The aim of the study was to develop and optimize the preparation of freeze-dried cytochrome C-containing liposomes (FD-Cyt-LS) using the Quality by Design (QbD) approach and to compare the stability of aqueous Cyt-LS and FD-Cyt-LS. In the case of freeze-dried Cyt-LS (FD-Cyt-LS), the influence of cryoprotectants type and concentration, as well as secondary drying temperature, which ensure integrity of liposomal membrane during lyophilization and preservation throughout the shelf life were studied. Lactose with a concentration of 6% was established to be the optimal type of cryoprotectant. At the lyophilization programme selected the most preferable secondary drying temperature value was 30°C. The necessity to choose the form of lyophilizate for solution preparation was shown with the example of stability evaluation of aqueous Cyt-LS and FD-Cyt-LS. During the observation period, aqueous Cyt-LS shows the growth of degradation products of free fatty acids and lysophosphatidylcholine (LPC) by 202.5% and 409.7% respectively, compared with the reference values, as well as the decrease of the content of phosphatidylcholine (РС) and dipalmitoylphosphatidylcholine (DPPG- Na) by 1.5 and 1.0%, respectively. At the same time, no significant changes in the composition of FD-Cyt-LS samples were observed.
The study of the lipid membrane charge effect when creating liposomes with oxaliplatin
(Національний фармацевтичний університет, 2016) Stadnichenko, O. V.; Krasnopolsky, Yu. M.; Yarnykh, T. G.
The fight against cancer diseases is one of the most urgent problems of modern pharmacy. One of the basic standards of treatment is surgery in order to remove tumours with chemotherapeutic agents for suppression of disease manifestations. One of the ways for reducing toxicity of cytostatics is their incorporation into liposomes – nanoparticles composed of the lipid bilayer surrounding the internal cavity with the aqueous medium. Oxaliplatin is a platinum-containing chemotherapeutic agent of the 3-rd generation used as monotherapy or in combination with other drugs. When creating liposomal drugs the beginning of the work is associated with the study of the composition of the lipid membrane. The aim of the work is to study the effect of the lipid membrane charge when creating liposomes with oxaliplatin. Four types of differently charged lipid membranes for liposomal oxaliplatin formulation have been tested. Liposomes were formed by the lipid layer method with further high pressure homogenization. As a criterion the encapsulation degree was used. The highest encapsulation degree has been determined in negatively charged liposomes with the lipid membrane modified by dipalmitoyl phosphatidylglycerol (DPPG).
Experiment planning at the pharmaceutical development of liposomal cytostatics
(Національний фармацевтичний університет, 2017) Stadnichenko, A. V.; Krasnopolsky, Yu. M.; Yarnykh, T. G.
At present, there is an increasing interest in developing new ways of drug delivery and targeted therapy, using nanotechnology and nanomaterials. Aim – to study the order of carrying out of pharmaceutical development of liposoms with cytostatics. Propose the scope of the experiment to optimize the planned quality indicators and technological parameters. Materials and methods. Analysis of normative documents, scientific literature and also the results of previous personal experimental studies, which became the basis for determining the methodology for the creation of liposomal drugs based on oxaliplatin and irinotecan. Lipids manufactured by Lipoid, Germany, were used to make liposomes. The lipid film was produced on a Buchi 210 rotary evaporator with a vacuum controller, at a residual pressure of 0.02 atm. For homogenization, a high pressure extrusion method was used, which was carried out on a Microfluidiser M-110P (Microfluidics, USA). Results and discussion. With the development of the pharmaceutical industry, there is a growing interest to the use of nanotechnology and nanomaterials. One of the practical implementation of nanotechnology is liposomes with cytostatics. Concentration of the active substance; pH and salt API; method of loading for API into liposomes; lipid to lipid ratio, lipids concentration; particle size and internal volume; lipid solubility in the step of lipid film preparation; stability testing of of finished products are factors that need to be studied and solved for the successful implementation of the development. Planning an experiment in the pharmaceutical development of liposomal oxaliplatin and liposomal irinotecan are complex studyes with using the principles of Quality by Design (QbD). Conclusions. The requirements of normative documentation for creating liposomal forms of medicinal products are considered. At pharmaceutical development it is necessary to use the complex approach as the majority of the put questions can not be solved separately. An experiment planning system for the pharmaceutical development of liposomal preparations of irinotecan and oxaliplatin is proposed.