Разработка рациональной лекарственной формы для активной фармацевтической субстанции (АФС) — одно из основных звеньев в цепи комплексных исследований по созданию лекарственного средства. Цель настоящего исследования — применение физико-химических и технологических приемов для создания различных парентеральных форм на основе практически нерастворимой в воде АФС — ЛХС-1269 и их сравнительное исследование по основным показателям качества, характерным для парентеральных ЛФ. В процессе работы были оценены составы и технологии получения трех моделей ЛФ на основе ЛХС-1269, показаны основные их преимущества и недостатки.

Производное индолокарбазола ЛХС-1269 обладает мультитаргетным механизмом воздействия на опухоль и блокирует васкулогенную мимикрию. По своим свойствам ЛХС-1269 относится к гидрофобным соединениям, что затрудняет получение инъекционной лекарственной формы (ИЛФ), необходимость создания которой определена в ходе ряда биологических опытов in vitro и in vivo. Цель настоящего исследования: выбор композиции вспомогательных компонентов для создания модели ИЛФ ЛХС-1269 с высокой противоопухолевой активностью и низкой токсичностью и разработка технологии ее получения. В ходе предварительных экспериментальных исследований разработаны состав модели ИЛФ ЛХС-1269, включающий спирт 95 %, Kollidon 17PF и полисорбат 80, и технология, позволяющая получить истинный раствор ЛХС-1269. Для исключения спирта из состава и для увеличения срока годности проведена лиофилизация выбранной модели.

Validation results for a spectrophotometric quantitative determination method for ormustine in a lyophilized dosage form are presented. The method is shown to be suitable for quantitative determination of ormustine with respect to parameters such as specificity, linearity, accuracy, repeatability, and intermediate precision.

A spectrophotometric assay for ormustine active ingredient for a liposomal dosage form with antitumor activity was selected and validated for specificity, linearity, accuracy, precision, and intermediate (intralaboratory) precision in order to ensure accurate and precise results. The obtained statistical characteristics were shown to satisfy acceptance criteria for the validation parameters given in domestic regulatory documentation. Ormustine was determined at 396 ± 2 nm because excipients in the dosage form did not absorb in this spectral region. The correlation coefficient for the linearity was >0.997. The relative error of the mean result was <1% for the accuracy. The confidence interval included 100%. The coefficient of variation for the precision and intermediate precision determinations was <1%. The studied method could be used in the range 80 – 120% of the nominal ormustine content in the liposomal dosage form.

An analog of somatostatin with hormonal and antitumor activity was synthesized. An optimum synthesis method was developed, yielding standard pharmaceutical somatostatin analog substance for preclinical and clinical studies. A quality control method was developed for inclusion in the draft manufacturer’s pharmacopeia monograph for pharmacological somatostatin analog substance.

Синтезирован аналог соматостатина, обладающий гормональной и противоопухолевой активностью. Разработан оптимальный метод синтеза, позволяющий получать стандартную фармацевтическую субстанцию аналога соматостатина для доклинических и клинических исследований. Разработаны методики контроля качества, которые будут включены в проект ФСП на фармацевтическую субстанцию аналога соматостатина.

The optimum composition of a stable lyophilized liposomal formulation of tiosens is established and includes lecithin, cholesterol, and PEG-2000-DSPE in a molar ratio of 1:0.22:0.002 and sucrose solution as a cryoprotector in a 1:9 lecithin:cryoprotector weight ratio. The effects of sonication, homogenization, and filtration on the quality of the liposomal preparation were studied. The results of preliminary studies lead to a conclusion about the satisfactory storage stability of the liposome form of tiosens.

Chromatographic and spectrophotometric methods for the qualitative and quantitative analysis of thermosensitive liposomes loaded with doxorubicin have been developed. Lipids and doxorubicin in thermosensitive liposomes have been separated by thin layer chromatography TLC) in two systems: (i) chloroform - methanol - ammonia (65:25:4) and (ii) chloroform - methanol - glacial acetic acid - water (25:15:4:2). The chromatographic determination of sucrose in thermosensitive liposomes has been performed in 1,2-dichloroethane - anhydrous acetic acid - methanol - water (10:5:3:2) or isopropyl alcohol - acetone - ether - water (7:7:2:4) systems. A spectrophotometric assay of doxorubicin in thermosensitive liposomes has been validated.

Freshly prepared liposome dispersions have a limited shelf-life, which hinders large-scale production with subsequent storage and sales to patients. This article addresses the development of a cifelin freeze-dried pegylated liposomal form. The effects of various cryoprotectors (glucose, sucrose, lactose, and mannitol) on the hydrophobic drug entrapment efficiency and its size change in the liposome membrane during storage were studied. The best results were obtained using sucrose with an optimum lipid:cryoprotector ratio of 1:1.4. Quality parameters such as vesicle size, pH value, and cifelin entrapment efficiency for the resulting freeze-dried liposomal dosage form remained unchanged during the studied storage time.

Выбран оптимальный состав и отработана технология получения липосомальной лекарственной формы лизомустина. С целью повышения стабильности лекарственной формы проведена лиофилизация липосомальной дисперсии с использованием криопротектора - 10 % раствора сахарозы. Для количественного определения содержания лизомустина в липосомах предложен метод спектрофотометрии при длине волны 230 нм. Включение лизомустина в свежеприготовленные липосомы составило 61,2 ± 0,4 %, размер частиц - 170 ± 20 нм.

The directed delivery anticancer drugs by immunoliposomal transport systems are one of the ways to realize the target strategies. The MUC-1 antigen expressed on the surface of the most common human cancer is one of the attractive targets for directed delivery of anticancer drugs. The purpose of present work was preparation anti-MUC-1 immunoliposomal construction loaded with doxorubicin and assessment their specific activity in vitro. Immunoliposomal constructions were prepared by reverse evaporation method using phospholipids, lipid-grafted PEG cholesterol and activated PNp-PEG3000 lipid with their subsequent loading by doxorubicin based on the formation of ammonium gradient between the internal and external aqueous phase of liposomes. The efficacy of doxorubicin encapsulation in immunoliposomes was 87-94 % the diameter of vesicles was 140 ± 5 nm. The high level of аntigen specificity and cytotoxic activity of this construction was obtained.

The optimal composition and production technique were identified for preparation of liposomal formulations of lisomustine. The stability of this formulation was increased by lyophilization of the liposomal dispersion using a cryoprotector, i.e., 10% sucrose solution. The lisomustine content in lysosomes was assayed by spectrophotometry at a wavelength of 230 nm. Uptake of lisomustine into freshly prepared liposomes amounted to 61.2 ± 0.4% and particle size was 170 ± 20 nm.

Fotoditazin, photosensitizer of second generation, is used in photodynamic therapy of tumors. In order to improve selectivity of drug accumulation in tumor tissue and improve the stability of the drug during storage, « Lyophilized liposomal fotoditazin for solution for injection 1,5 mg», a new liposomal drug form of fotoditazin, has been worked out in N. N. Blokhin Russian Cancer Research Center. This paper presents methods for identification of egg phosphatidylcholine, fotoditazin and methods of fotoditazin assay in the new lyophilized liposomal drug.

Photodithazine, a second-generation photosensitizer, is used in the photodynamic therapy of tumors. With the aims of improving the selectivity of its accumulation in tumor tissue and increasing its stability on storage, studies at the N. N. Blokhin Russian Oncological Scientific Center, Russian Academy of Medical Sciences, led to the creation of a new liposomal formulation of Photodithazine, “Liposomal Photodithazine, lyophilisate for preparation of solution for injection 1.5 mg.” We report here the development of a method for identifying egg phosphatidylcholine and Photodithazine and a method for assaying Photodithazine contents in the new lyophilized liposomal formulation.