Lyophilization of protein and peptide anticancer drugs is a promising method to create a stable and effective parenteral dosage forms. This article reviews the main problems encountered during lyophilization of protein and peptide drugs, and describes the best practices for solving these problems. The ways of optimization and improvement of the freeze-drying are given. Temperatures to improve the properties and prevent the destruction of the cake lyophilizatiе are suggested.

Dendritic cells play key role during tumorigenesis and immune response to it. They are able to uptake and present antigens to T cells, resulting in specific T cell mediated immune response. Furthermore, interaction between dendritic cells and other types of immune cells may boost cell-mediated and humoral immune response to cancer. Contrary to that, numerous tumor-derived factors may attract dendritic cells to neoplastic sites, causing impairment of their maturation, differentiation, and functional activity, resulting in deficiency of anti-tumor immune response or dendritic cell-mediated tolerance. Various factors within tumor microenvironment may either stimulate or inhibit dendritic cells and therefore need to be determined for improving efficacy of biotherapy utilizing dendritic cells. Meanwhile, recovery of dendritic cells functions in cancer patients remains one of primary aims for cancer immunotherapy. This review outlines main types of tumor-derived factors and their impact on dendritic cells in cancer.

Introduction. Immunochemotherapy is an obligate component in treatment of follicular lymphoma (FL). Monoclonal antibodies to CD20 antigen is the most well studied and frequently used drug. Chemotherapy department of Russian Cancer Research Center has a very good experience in FL treatment using rituximab. In the article we characterize and compare two methods of treatment of FL - polychemotherapy vs chemotherapy. Materials and methods. 286 patients with FL were included into analysis. In 125 (44 %) from 286 patients polychemotherapy was used as a first line of FL treatment. In 161 (56 %) patients rituximab was added to chemothrepy regimens. Analysis of survival and disease-free survival was done according to Kaplan-Meier. Analysis of treatment results has been done according to international criteria of treatment response in non-Hodgkin’s lymphomas. In statistical calculations SPSS program for Windows was used. Results. Responses to treatment were significantly better in immunochemotherapy group of patients, p = 0,002. In unifactorial analysis rituximab significantly improved both overall and disease-free sirvival. Taking in mind that groups of patients receiving chemotherapy and immunochemotherapy were not completely identical we performed multifactorial analysis, Coxs regression analysis. In that case (Cox regression analysis) significant advantages of immunochemotherapy were noted in improving disease-free survival in FL patients (р = 0,000); significant influence on overall survival was not noted (p = 0,44). Addition of rituximab did not improved overall and disease-free survival in a group (n = 88; 31 %) of refractory and early relapsing FL. Conclusion. Addition of rituximab to polychemotherapy in FL patients significantly improved treatment results. In multifactorial analysis immunochemotherapy was the most important prognostic factors of favourable prognosis of disease-free survival. In that analysis rituximab did not influenced on overall survival and on survival in refractory and early relapsing FL.

Introduction. So far in the literature there is no consensus on the relationship between the level of androgens and their metabolites in various biological fluids and tissues with the development of breast cancer. As suggested by many researchers, determining the level of androgenetic receptors (AR) in the tumor in patients with breast cancer are able to access a number of promising directions in the study of prognosis of the disease and the search for new approaches to endocrine therapy of different molecular subtypes of breast cancer. Objective. To evaluate the clinical and prognostic significance of the immunohistochemical expression of AR in different molecular types of breast cancer. Materials and methods. In the present work we have studied the expression of AR in tumor tissue in patients of different molecular types of breast cancer. Results. The risk of death in patients with AR-positive breast cancer is 88 % lower than in patients with AR-negative tumors. Conclusion. It was concluded that the study of the expression of AR, is a promising direction not only in the assessment of the prognosis of the disease, but when searching for additional approaches to the treatment of breast cancer.

Introduction. In patients with neuroendocrine neoplasms it is necessary to search for new prognostic factors to identify a subgroup of patients who will benefit from drug treatment. Aim. The aim of the study was an optimization of Aranoza-based therapy in patients with neuroendocrine neoplasms by using both morphological and clinical factors as predictors of the treatment efficacy. Materials and methods. A total of 122 patients were enrolled and assessed for radiological response (RECIST 1.0) and progression-free survival. Results. Aranoza and its combinations had similar efficacy. The localization of metastases only in the liver, less than 3 metastatic zones, more than 6 courses of Aranoza-bazed chemotherapy, well-differentiated tumors, absence of doses reduction of Aranoza and temozolomide in anamnesis were predictors of the disease control rate. Progression-free survival was higher in patients with disease control rate (objective response + stabilization more than 6 months), Ki-67 < 20 %, well-differentiated tumors, more than 6 courses of Arano-za-based chemotherapy, objective biochemical response, less than 3 metastatic zones and absence of O6-alkylguanine-DNA alkyl-transferase expression. Conclusion. Future analysis of prognostic factors in patients treated with alkylating agents is needed.

Introduction. An increase in patients with diabetes and a large number of amputations of limbs make the search for new treatments for these patients. Materials and methods. The study included 17 patients with complicated forms of diabetes. Patients were studied in the bone marrow immediately after surgery - the amputation of the limb. The number of hematopoietic stem cells and mesenchymal stem cells using specific markers were determined in the laboratory. The immunohistochemical characteristics of bone marrow were carried out for all samples. Results and conclusion. The obtained results indicate the biological value of bone marrow removed with amputation of the lower limb of the patient with complicated diabetes mellitus. The ease of obtaining bone marrow makes the procedure approachable in surgical practice. The presence of hematopoietic and mesenchymal stem cells provides the opportunity to use them in autologous cell therapy for patient.

Introduction. Creation of new orthotopic models of tumor growth (surgical orthotopic implantation, SOI) promotes pre-clinical studies of anti-metastatic functional potential of the drugs including those which aim to treat advanced clear-cell renal carcinoma with the use of patient’s tumor or metastasis sample for implantation material. The use of subcutaneous (s. c.) xenograft contributes to SOI improvement and extends experimental opportunities. The collection of tumor strains of N.N. Blokhin Russian Cancer Research Center that includes s. c. xenograft RPoch1, which was characterized by transplantation, pathological, and immunological parameters and in terms of sensitivity towards classical antitumor cytostatics, presents the base for obtaining an orthotopic model for multifunctional usage in pre-clinical studies of specific medicines. Objective. Preparation and obtaining an orthotopic model of human clear-cell renal carcinoma with the implant of subcutaneous xenograft RPochl. Materials and methods. The study used s. c. RPoch1 and immunodeficient female mice Balb/c nude on the N. N. Blokhin Russian Cancer Research Center breeding. Transplantation characteristics were defined during implant passages into kidney parenchyma. The effectiveness of orthotopic implantation was estimated in the dynamics according to kinetic, pathological, morphological, and immunological parameters by light microscopy, flow cytometry, monoclonal antibodies (mAb, Caltag Laboratories) against corresponding antigens. Sensitivity to specific drugs was assessed against inhibitors of tyrosine kinases according to standard parameters (T/C <42 %, total regressions). Results. After orthotopic implantation of RPochl on days 7, 14 and 21 the tumor lesions were registered with the size of 5.0 ± 1.0 mm³, 215.0 ± 88.0 mm³ and 608.0 ± 157.0 mm³. Histological examination showed that the tumor had separated connective tissue layers of solid areas of large polymorphic cells with mitosis (10-15 in a lesion) and single areas with necrosis on day 21. The 1.5-fold enhanced expression of NK1.1 in mice on day 14 and 3.3-fold increase of CD8a expression demonstrate the tendency to develop immune suppression induced by growing tumor. The results showed high sensitivity of RPoch1 to sorafenib and sunitinib (T/C_mn = 2-30 %). Conclusion. The authors obtained an orthotopic xenograft of human clear-cell renal carcinoma RPoch1 with 100 % transplantation potential, reproducible biological characteristics including immunological parameters and sensitivity to inhibitors of tyrosine kinases, which can be used for pre-clinical studies of various agents including immunological or different combinations of antitumor drugs against this cancer type.

Introduction. Glucocorticoids are the important component of combined chemotherapy of blood cancer. Therapeutic effects of glucocorticoids is realized via activation of glucocorticoid receptor transrepression, the development of side effects is associated with transactivation. We demonstrated earlier that compound belonging the class of selective glucocorticoid receptor agonists, CpdA, selectively induced transrepression in blood cancer cells. CpdA represents a mixture of two enantiomers, which can differ in interaction with the receptor. Aim. The main aim of present study was to synthesize CpdA enantiomers and to evaluate their biological properties. Materials and methods. Synthesis was carried out based on Sharpless dihydroxylation; anti-tumor activity in vitro was evaluated by antiproliferative and pro-apoptotic effects. Ligand properties were estimated by PCR-analysis of glucocorticoid- and NF-kB-dependent genes expression. Results and conclusions. We demonstrated that CpdA enantiomers revealed anti-tumor activity in vitro and did not induce transactivation. Moreover, S-enantiomer of CpdA in the most tests demonstrated more pronounced activity and is more perspective molecule for future studies in vivo.

Introduction. Until now, the search continues for new anticancer drugs with high efficacy and low toxicity profile. One promising class of anticancer agents that meet these requirements are histone deacetylase inhibitors (HDAC), in particular, sodium phenylbutyrate (PB). Aim. Experimental estimate of the dose and time dependence of the antitumor activity, and tolerability during prolonged oral administration of PB in combination with 5-fluorouracil (5-FU) on Ehrlich carcinoma model. Materials and methods. The animals were divided into 6 groups (n = 20 mice): 1^st (control group) - received normal drinking water, 2^nd - PB (800 mg/kg), an aqueous solution instead of drinking water, 3^th - 5-FU (200 mg/kg), as the maximum tolerated dose (MTD), 4^th - PB +5-FU (800, 200 mg/kg), 5^th - 5-FU(100 mg/kg) intraperitoneally, once, and 6^th - PB + 5-FU (800,100 mg/kg). The drugs were administered sequentially 48 hours after inoculation for 14 days. Results. The most pronounced inhibition of tumor growth was observed in the combined treatment on the 14^th day after inoculation in group PB + 5-FU (200 mg/kg) and PB + 5-FU (100 mg/kg), tumorgrowth inhibition (TGI) = 92-96 % and 83-90 % respectively, compared to controls. Comparison of combined treatment with monotherapy showed a significant increase in TGI = 63-72 %, and with respect to PB and 54-57 % relative to 5-FU, which indicating of synergistically antitumor effect, which persisted after treatment on level of TGI = 72-87 % (p < 0.05), which persisted for 21 days on level of TGI = 72-87%. Prolonged oral administration PB in combination with 5-FU significantly improved tolerance and increased lifespan of the animals at 33 %, which was 1.5 times more, compared with monotherapy. Conclusions. These data allow to conclude that a rational combination of HDAC with cytotoxic agent is able to overcome the resistance mechanism of tumor cells and lead to a synergistic antitumor effect and lower toxicity.

The aim of this investigation was to evaluate the life-span and physical status (weight, hair character, motor activity) of CBA inbred mice with high frequency of spontaneous hepatocarcinomas using multiphytoadaptogene complex (MPAC) preventive administration in early postnatal ontogenesis. Materials and method. MPAC is a herbal formula composed of forty plants water-ethanol extracts components including adaptogenes Panax ginseng, Eleutheroccosus senticosus, Rhodiola rosea. Anti-tumour, anti-oxidant, anti-mutagenic, immunomodifying activities of MPAC was demonstrated in previous studies. The study was carried out on 131 males of CBA mouse strain. The experimental mice received 10 % MPAC solution in drinking water during the first month of life including the final time period of liver differentiation. Motor activity was determined by a test of the «open field» automatically using Opto-Varimex-3 (Columbus Instruments, USA). The average lifespan was evaluated. Statistical analysis was performed with the program STATISTICA 6.0. Survival median was determined by the Kaplan-Meier method. Results. MPAC preventive use in early postnatal ontogenesis promotes mouse longevity increase for 17,1 % (p < 0,001), longevity median -for 25,6 % (p < 0,001). Physical status of experimental animals using MPAC administration had an advantage over control mice. Conclusion. So, data obtained demonstrate nontoxic MPAC administration in early ontogenesis promotes significant life-span increase, keeps the body weight, good coat state, motor activity as well as prevents hepatocarcinomas formation.

Objective. The purpose of the research is the experimental study of the possibility of correction of disorders of lipid metabolism, lipid peroxidation and antioxidant protection (POL-AOP), liver structure on the model of alimentary hyperlipidemia in mice. Materials and methods. Fucoidan, a sulfated polysaccharide with a molecular weight of 160 kDa, derived from brown algae Fucus evanescens. The alimentary hyperlipidemia model was reproduced in mice. The biochemical parameters of lipid metabolism, state of POL-AOP system and the liver structure by MRT were studied. Results. We revealed the ability of fucoidan at per os administration to animals to normalize the key parameters of lipid metabolism, indicators of POL-AOP system, liver anatomic-topographic structure. Conclusion. The ability of fucoidan to correct these parameters allows us to consider it as a basis for the development of new biological medicines for the treatment of atherosclerotic disorders.