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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"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.
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.
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.
The study of liophilization parameters in the liposomal irinotecan development
(Національний фармацевтичний університет, 2017) Stadnichenko, O. V.; Krasnopolsky, Yu. M.; Yarnykh, T. G.
The creation of the liposomal irinotecan is one of the main ways to reduce toxicity and increase the effectiveness of chemotherapy. Lyophilization makes it possible to obtain a product with a guaranteed stability of the size and encapsulation efficiency. Aim. To optimize the content of the cryoprotector in the liposomal irinotecan, and develop lyophilization parameters to produce liposomes with the maximum encapsulation of irinotecan in them, alongside while maintaining the nanosize. Materials and methods. Egg phosphatidylcholine from Lipoid (Germany) was used for preparation of liposomes. Lyophilization was carried out in a Quarco device (PR China). The encapsulation degree was determined on a Shimadzu LC-20 instrument (Japan) by HPLC method developed earlier. Results and discussion. The optimal content of the cryoprotector – trehalose dihydrate has been studied. It has been found that the optimal content of trehalose dihydrate is 8 % (w/w). The modes of the product lyophilization have been studied. The secondary drying temperature in the range of 10-20 °C has been determined. At the secondary drying temperature of 10 °C the residual moisture content was 5-8 %, which was beyond the target range. At 20 °C the water content in the lyophilizate was 0.5-0.8 %, the loss of encapsulation was up to 20 %. The mode of drying at 15 °C was optimal, while the residual water content in the lyophilizate was 1.5-2.8 %, the loss of encapsulation was 13 %, the size of the liposomes after lyophilization and rehydration did not change significantly compared to the initial one. Сonclusions. As a result of the studies, liposomes with irinotecan have been obtained. The content of trehalose dihydrate as a cryoprotector in the range of 4-10 % has been studied. It has been shown that the optimum content of trehalose dihydrate is 8 % (w/w); moreover, the encapsulation decrease in lyophilization is 13 %. The modes of the liposomal irinotecan lyophilization have been studied at the final drying temperature of 10, 15 and 20 °C. It has been found that the optimum final drying temperature is 15 °C.
Study of the composition of cryoprotector and technological regime in liophilization of liposomes with oxaliplatinum
(НВП ПП "Технологічний Центр", 2017) Stadnichenko, A. V.; Krasnopolsky, Yu. M.; Yarnykh, T. G.
Lyophilization is one of the most prospective and technologically logical methods for preserving the structure of nanobiotechnological products. Liposomes with oxaliplatin were obtained, and a screening experiment was performed to select a cryoprotectant. Aim. The aim of the research is to obtain liposomes with oxaliplatin, determine the type of cryoprotectant and its quantity, study the parameters of lyophilisation to obtain the product with maximum encapsulation of oxaliplatin, with the save of the size of liposomes in the nano diapason, and the optimum residual moisture content. Methods. Lyophilization was carried out in a Quarco lyophilizer (PRC). The liposomal form of oxaliplatin was obtained by the method of “passive” encapsulation in combination with the ion sorption method. Results. Lactose, sucrose, maltose and trehalose dihydrate were studied as cryoprotectants. As the most perspective, was chosen - trehalose dihydrate. The optimal concentration of trehalose dihydrate in the liposomal form of oxaliplatin was researched at 8 % by weight. Also was optimized the program of freeze-drying. Primary drying with a duration of 1740 minutes was not sufficient. When the program was increased for 300 min, up to 2040 min, the values of the loss of encapsulation rate of 8 %, from 65 % before drying, to 57 % after drying were obtained. This is a good indicator, which shows an effectiveness of cryoprotectant and a rational program of freeze-drying. The size of the liposomes after drying was 112 nm, the residual moisture content, measured by K. Fischer’s method, was 2.3 %, which is within the scope of the target range. Conclusion. A technology for obtaining liposomes with encapsulated oxaliplatin has been proposed and screening studies have been carried out to determine the optimal cryoprotectant. It is proposed to use as a cryoprotector trehalose dihydrate as the most perspective. The effect of different content of trehalose dihydrate on the degree of encapsulation of oxaliplatin in liposomes was studied. It was found= that the optimal cryoprotectant concentration in the preparation is 8 % (by mass). Technological parameters of the lyophilization process of liposomes with oxaliplatin have been developed: drying time and freezing temperature. The decrease in the incorporation of oxaliplatin into liposomes during lyophilisation did not exceed 8.0 % with a residual water content of about 2.3 %. The size of liposomes after lyophilization in the nanoscale is 112 nm.