Subject(s)
Bipolar Disorder/genetics , Diacylglycerol Kinase/genetics , Polymorphism, Single Nucleotide/genetics , Adult , Female , Gene Frequency , Genotype , Humans , Italy , Male , Middle AgedABSTRACT
A reversed-phase high-performance liquid chromatography (HPLC) method with diode-array detection for the quantification of several human salivary peptides is described. Sample pretreatment consisted of the acidification of whole saliva by phosphate buffer. This treatment produced precipitation of mucins, alpha-amylases and other high-molecular-mass salivary proteins, simultaneous inhibition of intrinsic protease activities and reduction of sample viscosity. Direct HPLC analysis by diode-array detection of the resulting acidic sample allowed one to quantify histatin 1, histatin 3, histatin 5, statherin, as well as uric acid, in normal subjects. Moreover, the groups of peaks pertaining to proline-rich proteins and cystatins were tentatively identified. The method can be useful in assessing the concentration of salivary peptides from normal subjects and from patients suffering oral and/or periodontal diseases.
Subject(s)
Chromatography, High Pressure Liquid/methods , Phosphopeptides/analysis , Proteins/analysis , Salivary Proteins and Peptides/analysis , Acids , Buffers , Histatins , Humans , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
A series of indolo[3,2-c]cinnoline derivatives was prepared and tested to evaluate their biological activity. Most of them inhibited the proliferation of leukemia, lymphoma and solid tumor-derived cell lines at micromolar concentrations, whereas none of the compounds were active against HIV-1. With the exception of 7g, all title compounds showed antibacterial activity against gram-positive bacteria, being up to 200 times more potent than the reference drug streptomycin. Some of the indolo[3,2-c]cinnolines were also endowed with good antifungal activity, particularly against Criptococcus neoformans.
Subject(s)
Anti-Infective Agents/pharmacology , Antifungal Agents/pharmacology , Antineoplastic Agents/pharmacology , Indoles/pharmacology , Phthalazines/pharmacology , Anti-Bacterial Agents , Anti-Infective Agents/chemistry , Antifungal Agents/chemistry , Antineoplastic Agents/chemistry , Cell Division/drug effects , Cryptococcus neoformans/drug effects , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Humans , Indoles/chemistry , Microbial Sensitivity Tests , Phthalazines/chemistry , Tumor Cells, CulturedABSTRACT
A series of 2-triazenothiophene derivatives was prepared and tested to evaluate their biological activity. Two compounds inhibited the proliferation of leukemia, lymphoma and solid tumor-derived cell lines at micromolar concentrations, whereas none of the compounds were active against HIV-1. Compound 3c inhibited DNA, RNA and protein synthesis, and was also effective against KB cells resistant to etoposide and vincristine. The compounds were inactive against fungi and bacteria.
Subject(s)
Antineoplastic Agents/pharmacology , Thiophenes/pharmacology , Triazenes/pharmacology , Anti-Bacterial Agents , Anti-HIV Agents/pharmacology , Anti-Infective Agents/pharmacology , Antineoplastic Agents/chemical synthesis , Bacteria/drug effects , Carcinoma/pathology , Cytopathogenic Effect, Viral/drug effects , Drug Evaluation, Preclinical , Drug Resistance, Neoplasm , Drug Screening Assays, Antitumor , Fungi/drug effects , HIV-1/drug effects , HeLa Cells/drug effects , Humans , KB Cells/drug effects , Leukemia/pathology , Lymphoma/pathology , Melanoma/pathology , Structure-Activity Relationship , Thiophenes/chemical synthesis , Triazenes/chemical synthesis , Tumor Cells, Cultured/drug effectsABSTRACT
A new set of phthalein derivatives stemming from the lead compound, phenolphthalein, were designed to specifically complement structural features of a bacterial form of thymidylate synthase (Lactobacillus casei, LcTS) versus the human TS (hTS) enzyme. The new compounds were screened for their activity and their specificity against TS enzymes from different species, namely, L. casei (LcTS), Pneumocystis carinii (PcTS), Cryptococcus neoformans (CnTS), and human thymidylate synthase (hTS). Apparent inhibition constants (Ki) for all the compounds against LcTS were determined, and inhibition factors (IF, ratio between the initial rates of the enzymatic reaction in the presence and absence of each inhibitor) against each of the four TS species were measured. A strong correlation was found between the two activity parameters, IF and Ki, and therefore the simpler IF was used as a screening factor in order to accelerate biological evaluation. Compounds 5b, 5c, 5ba, and 6bc showed substantial inhibition of LcTS while remaining largely inactive against hTS, illustrating for the first time remarkable species specificity among TSs. Due to sequence homology between the enzymes, several compounds also showed high activity and specificity for CnTS. In particular, 3-hydroxy-3-(3-chloro-4-hydroxyphenyl)-6-nitro-1H, 3H-naphtho[1,8-c,d]pyran-1-one (6bc) showed an IF < 0.04 for CnTS (Ki = 0.45 microM) while remaining inactive in the hTS assay at the maximum solubility concentration of the compound (200 microM). In cell culture assays most of the compounds were found to be noncytotoxic to human cell lines but were cytotoxic against several species of Gram-positive bacteria. These results are consistent with the enzymatic assays. Intriguingly, several compounds also had selective activity against Cr. neoformans in cell culture assay. In general, the most active and selective compounds against the Gram-positive bacteria were those designed and found in the enzyme assay to be specific for LcTS versus hTS. The original lead compound was least selective against most of the cell lines tested. To our knowledge these compounds are the first TS inhibitors selective for bacterial TS with respect to hTS.
