piperidines and diperodon

SMYD3 is a multifunctional epigenetic enzyme with lysine methyltransferase activity and various interaction partners. It is implicated in the pathophysiology of cancers but with an unclear mechanism. To discover tool compounds for clarifying its biochemistry and potential as a therapeutic target, a set of drug-like compounds was screened in a biosensor-based competition assay. Diperodon was identified as an allosteric ligand; its R and S enantiomers were isolated, and their affinities to SMYD3 were determined (K

Topics: Allosteric Site; Binding Sites; Cell Line, Tumor; Drug Evaluation, Preclinical; Histone-Lysine N-Methyltransferase; HSP90 Heat-Shock Proteins; Humans; Kinetics; Ligands; Molecular Dynamics Simulation; Piperidines; Protein Binding; Stereoisomerism

The transcription factor C/EBPα is required for granulocytic differentiation of normal myeloid progenitors and is frequently inactivated in acute myeloid leukemia (AML) cells. Ectopic expression of C/EBPα in AML cells suppresses proliferation and induces differentiation suggesting that restoring C/EBPα expression/activity in AML cells could be therapeutically useful. Unfortunately, current approaches of gene or protein delivery in leukemic cells are unsatisfactory. However, "drug repurposing" is becoming a very attractive strategy to identify potential new uses for existing drugs. In this study, we assessed the biological effects of candidate C/EBPα-mimetics identified by interrogation of the Connectivity Map database. We found that amantadine, an antiviral and anti-Parkinson agent, induced a monocyte-macrophage-like differentiation of HL60, U937, Kasumi-1 myeloid leukemia cell lines, as indicated by morphology and differentiation antigen expression, when used in combination with suboptimal concentration of all trans retinoic acid (ATRA) or Vit D3. The effect of amantadine depends, in part, on increased activity of the vitamin D receptor (VDR), since it induced VDR expression and amantadine-dependent monocyte-macrophage differentiation of HL60 cells was blocked by expression of dominant-negative VDR. These results reveal a new function for amantadine and support the concept that screening of the Connectivity Map database can identify small molecules that mimic the effect of transcription factors required for myelo-monocytic differentiation.

Topics: Amantadine; Antigens, CD; Antineoplastic Agents; CCAAT-Enhancer-Binding Proteins; Cell Cycle; Cell Differentiation; Cell Proliferation; Gene Expression; HL-60 Cells; Humans; Hypoxanthine Phosphoribosyltransferase; K562 Cells; Leukemia, Myeloid, Acute; Macrophages; Piperidines; Protein Interaction Maps; Receptors, Calcitriol; Tamoxifen; Tretinoin

Label free time-lapse microscopy has opened a new avenue to the study of time evolving events in living cells. When combined with automated image analysis it provides a powerful tool that enables automated large-scale spatiotemporal quantification at the cell population level. Very few attempts, however, have been reported regarding the design of image analysis algorithms dedicated to the detection of apoptotic cells in such time-lapse microscopy images. In particular, none of the reported attempts is based on sufficiently fast signal processing algorithms to enable large-scale detection of apoptosis within hours/days without access to high-end computers. Here we show that it is indeed possible to successfully detect chemically induced apoptosis by applying a two-dimensional linear matched filter tailored to the detection of objects with the typical features of an apoptotic cell in phase-contrast images. First a set of recorded computational detections of apoptosis was validated by comparison with apoptosis specific caspase activity readouts obtained via a fluorescence based assay. Then a large screen encompassing 2,866 drug like compounds was performed using the human colorectal carcinoma cell line HCT116. In addition to many well known inducers (positive controls) the screening resulted in the detection of two compounds here reported for the first time to induce apoptosis.

Topics: Antibiotics, Antineoplastic; Apoptosis; Caspase 3; Caspase 7; HCT116 Cells; High-Throughput Screening Assays; Humans; Microscopy; Mitomycin; Naphthoquinones; Organic Chemicals; Piperidines; Staining and Labeling; Time-Lapse Imaging

The chiral stationary phase on the base of teicoplanin and the polar-organic mobile phase methanol/acetonitrile/acetic acid/triethylamine 45/55/0.3/0.2 was used for the separation of diperodon enantiomers. The developed method was suitable to determine the enantiomers in blood serum up to 0.5 microg/ml. The degradation of diperodon enantiomers was studied in serum by an in vitro method and the experimental rate constants were determined.

Topics: Chromatography, High Pressure Liquid; Humans; Piperidines; Stereoisomerism; Teicoplanin

An on-line coupled HPLC system is described for the determination of the enantiomers of diperodon in blood serum. The method involves three steps: (i) off-line preconcentration and clean-up, (ii) separation of the diperodon enantiomers from the matrix components on a reversed-phase stationary phase, and (iii) separation of the racemate from the reversed-phase column on a teicoplanin chiral stationary phase. The method is suitable for simultaneous determination of both enantiomers in serum up to 0.5 microg/ml. The degradation of diperodon enantiomers was studied in serum by an in vitro method and the experimental rate constants were determined. The enantiomeric hydrolysis rates and half-lives for diperodon in serum are different.

Topics: Animals; Chromatography, Liquid; Enzymes; Hydrolysis; Kinetics; Piperidines; Rabbits; Stereoisomerism

Topics: Adult; Dermatitis, Atopic; Dermatitis, Contact; Humans; Male; Piperidines