ABSTRACT
The retention of 16 quinoline and tetrahydroquinoline derivatives was investigated under liquid chromatography conditions using porous graphitized carbon (PGC), octadecyl silica (ODS) and hypercrosslinked polystyrene (HCLP) stationary phases. For most of the analytes, retention on PGC was greater than on ODS, while retention on HCLP was even greater than on both ODS and PGC. The non-linearity of retention dependencies on acetonitrile content in the eluent was observed for compounds containing carboxy, hydrazo and methoxy groups. The relationships between quinolines structure and their retention factors were investigated. It was found that sorption on different sorbents correlated with different descriptors. Retention on ODS was found to be highly correlated with lipophilicity only while on HCLP it depended on both lipophilicity and polarizability of the sorbates. The feature of PGC was a good correlation of retention factors with topological and geometrical parameters. Additionally, ability of PGC to form OH π bonds with hydroxymethylquinolines was found. The observed regularities allow one to rationally optimize the chromatographic analysis of structurally similar compounds, which is very important for bioactive substances.
ABSTRACT
The aim of our study was to investigate relationships between quinoline derivatives structure and their retention under reversed-phase liquid chromatography conditions. Retention factors of quinolines were experimentally measured and various geometrical and physicochemical parameters representing analytes molecular structure were calculated. Equations connecting chromatographic data with computed characteristics for the set of 17 investigated compounds were constructed. It was shown that the most precise dependencies include combination of physico-chemical and geometrical parameters.
Subject(s)
Chromatography, High Pressure Liquid , Quinolines/chemistry , Chromatography, Reverse-Phase , Quantitative Structure-Activity Relationship , Quinolines/isolation & purificationABSTRACT
We have previously described designing of polyepitope immunogens TBI and TCI, to stimulate the humoral and cellular immune responses to HIV-1. Here, immunogens TBI and TCI were used to create new vaccine construct named CombiHIVvac (Combined HIV-1 vaccine). CombiHIVvac is a virus-like particles (VLP) containing the DNA vaccine pcDNA-TCI as a core encapsulated within a spermidine-polyglucin-TBI conjugate. The immunogenic and toxic properties of the candidate vaccine CombiHIVvac have been studied. CombiHIVvac induces a strong humoral and CTL responses in mice; the antibodies are highly specific and are able to neutralize HIV-1 in vitro. Preclinical study demonstrated that CombiHIVvac does not cause long-term changes in physiological, biochemical and morphological parameters in immunized animals and thus can be recommended for clinical trials.
Subject(s)
AIDS Vaccines/immunology , Epitopes/immunology , HIV-1/immunology , Vaccines, DNA/immunology , Vaccines, Virosome/immunology , AIDS Vaccines/adverse effects , AIDS Vaccines/chemistry , Animals , Blotting, Western , Cells, Cultured , Cytokines/biosynthesis , Enzyme-Linked Immunosorbent Assay , Epitopes/genetics , HIV Antibodies/blood , Humans , Lymphocytes/immunology , Mice , Mice, Inbred BALB C , Models, Animal , Neutralization Tests , Vaccines, DNA/adverse effects , Vaccines, DNA/chemistry , Vaccines, Virosome/adverse effects , Vaccines, Virosome/chemistryABSTRACT
A synthetic T cell immunogen (TCI) has been designed as a candidate DNA-based vaccine against Human immunodeficiency virus (HIV)-1 using cytotoxic T lymphocytes (CD8(+) CTL) and T-helper lymphocytes (CD4(+) Th) epitopes retrieved from the Los Alamos HIV Molecular Immunology Database. The protein 392 amino acids in length contains about eighty CTL-epitopes, many of which are overlapping and are totally restricted by ten different HLA class I molecules. To be able to detect CTL responses induced by a DNA vaccine in experimental animals, additional epitopes, restricted by mouse and Macaque rhesus major histocompatibility complex (MHC) class I molecules, were included in the target immunogen. The gene encoding the TCI protein was assembled, cloned into vector plasmids and expressed in a prokaryotic and a eukaryotic system. The presence of HIV-1 protein fragments in the immunogen structure was ascertained by ELISA and immunoblotting using panels of HIV-1-positive sera and monoclonal antibodies to p24. It has been demonstrated that DNA vaccine can induce both specific T cell responses (CTL and blast transformation) and specific antibodies in mice immunized with pcDNA-TCI.
Subject(s)
AIDS Vaccines/genetics , AIDS Vaccines/immunology , Epitopes/genetics , Epitopes/immunology , HIV Antigens/genetics , HIV Antigens/immunology , HIV-1/genetics , HIV-1/immunology , T-Lymphocytes, Cytotoxic/immunology , Animals , Antibody Specificity , Base Sequence , Cell Division , DNA, Viral/genetics , DNA, Viral/immunology , Drug Design , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Genetic Engineering , HIV Antibodies/analysis , HIV Antibodies/biosynthesis , Humans , Immunochemistry , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Plasmids/immunology , Vaccines, DNA/immunologyABSTRACT
Two systems have been examined for delivery of DNA-vaccine encoding a HIV-1 polyepitope CTL-immunogen (TCI). One is intended for i.m. injection and is in the form of an artificial virus like particle containing eukaryotic expression plasmid pcDNA-TCI encapsulated within a spermidine-polyglucin conjugate. The other is intended for mucosal immunization and is based on attenuated Salmonella typhimurium strain 7207, which can deliver pcDNA-TCI directly into professional antigen-presenting cells (APC). After immunization, the artificial VLP and recombinant Salmonella induced an enhanced HIV specific serum antibody, proliferative and CTL responses compared to those induced by naked pcDNA-TCI. The most significant responses were produced when pcDNA-TCI was delivered by Salmonella.
