Inhibition of DNA topoisomerase I and growth inhibition of human cancer cell lines by an oleanane from Junellia aspera (Verbenaceae).

DNA topoisomerases and DNA polymerases are enzymes that play a crucial role in DNA metabolism events such as replication, transcription, recombination, and chromosome segregation during mitosis. Thus, DNA topoisomerases and DNA polymerases inhibitors could be expected to have antitumor effects. Naturally occurring triterpenoids isolated from Junellia aspera (Gillies & Hook; Moldenke) (Verbenaceae) were assayed for human DNA topoisomerase I and Taq DNA polymerase inhibitory activities. Maslinic acid (2) and its diacetyl derivative (7) showed human DNA topoisomerase I inhibitory activity with IC50 values in the range of 76-80 microM and growth inhibition against various human solid tumour cell lines with GI50 values in the range of 5-18 microM. The triterpene frames could be used for screening new inhibitors of the enzyme, and computer-simulated drug design using the frame and pocket structure of enzyme may in theory be a possible approach to develop new inhibitors.

Surface plasmon resonance investigations of human epidermal growth factor receptor 2.

This investigation utilizes surface plasmon resonance (SPR) spectroscopy to detect and quantify human epidermal growth factor receptor 2 (HER-2), an oncogene product that is over-expressed in some aggressive forms of breast cancer. Specifically, the HER-2 trans-membrane protein p185 and its extra cellular fragment p105 are analytes targeted in this work by using a gold-based biosensor slide on which an anti-HER-2 antibody has been immobilized by attachment to Protein G that is fixed to the gold film. A detection limit of > or =11 ng/mL for p185 resulted when trastuzumab was used as the anti-HER-2 antibody on the biosensor slide. Experiments with semi-purified p105 revealed that it binds weakly and reversibly to trastuzumab, therefore complicating its detection and quantification. Results of studies that reacted a 13-amino-acid peptide (PP13) from the HER-2 kinase domain with its specific antibody were critically different than p185 and p105 studies. Spectral analysis of the reflectivity at constant bulk buffer refractive index revealed a progressive negative SPR shift over time. A negative shift suggests that a loss of protein mass from the anti-PP13 antibody-Protein G biosensor is occurring. Several possibilities that may explain these negative SPR shifts are discussed.

Synthesis and in vitro cytotoxicity of long chain 2-amino alcohols and 1,2-diamines.

The synthesis of enantiomerically pure unsaturated long chain 1,2-diamines and amino alcohols was carried out starting from the corresponding non-natural alpha-amino acids. The in vitro cytotoxicity of the compounds prepared was evaluated against six solid tumor cell lines (A2780, H322, LL, WiDr, C26-10 and UMSCC-22B). Free 1, 2-diamines proved to be the most active compounds exhibiting IC50 values between 2.0 mM and 3.3 mM.

Double cationic propargylation: from linear to polycyclic ethers.

[reaction: see text]. The trapping of cations generated from Co2(CO)6-bispropargylic alcohols provided diethers in good yield. The procedure is also valid when two vicinal acetylenes are present. The methodology can be applied to the synthesis of symmetrical or unsymmetrical linear or cyclic propargylic ethers. The use of substrates with a stereochemically defined secondary nucleophilic alcohol provided cyclic ethers with a high degree of stereocontrol.

Beta-hydroxy-gamma-lactones as chiral building blocks for the enantioselective synthesis of marine natural products.

The enantioselective synthesis of trans-(+)-laurediol, (2S,3S,5R)-5-[(1R)-1-hydroxy-9-decenyl]-2-pentyltetrahydro-3-furanol, and (2S,3S,5S)-5-[(1S)-1-hydroxy-9-decenyl]-2-pentyltetrahydro-3-furanol are described. In addition, a formal synthesis of trans-(-)-kumausyne is also developed. All the synthetic procedures have in common the use of enantiomerically enriched beta-hydroxy-gamma-lactones, easily available by Sharpless asymmetric dihydroxylation (AD) from the suitable beta,gamma-unsaturated ester. The use of Katsuki-Sharpless asymmetric epoxidation (AE) as an additional enantioselective reaction provides cyclic compounds of high enantiomeric purity.

Synthesis, in vitro cytotoxicity and in vivo anti-inflammatory activity of long chain 3-amino-1,2-diols.

The synthesis of long chain 3-amino-1,2-diols was carried out based on Sharpless asymmetric epoxidation of long chain allylic alcohols and regioselective nucleophilic ring opening by azido group. The in vitro cytotoxicity of the compounds prepared was evaluated against six solid tumor cell lines (A2780, H322, LL, WiDr, C26-10, UMSCC-22B). Free 3-amino-1,2-diols exhibited IC50 values between 1.45 microM and 32 microM. These compounds also presented interesting inhibition of carrageenin-induced paw edema in rats (85.3% - 79.6% at a concentration of 0.15 mmol/kg).

Stability indicating high performance liquid chromatography methods for 5-fluorouridine in aqueous solution.

Two methods of reversed-phase high performance liquid chromatography (RP-HPLC) were used to detect and quantify the degradation of 5-fluorouridine (CAS 316-46-1, 5-FUR) in aqueous solution. The 1H-NMR technique was used for identifying the degradation products. Several assays were performed from different aqueous solutions of 5-FUR. The best results (optimal resolution, reproducibility and rapidity) were achieved by using Resolve C-18 as stationary phase and a mix (96:4) of 50 mmol/l ammonium dihydrogen phosphate (adjusted to pH 3.5 with phosphoric acid) and acetonitrile as mobile phase. Despite the 5-FUR hydrolysis to 5-fluorouracil (5-FU) and D-ribose under moderate conditions in unlikely, due to high stability of the nucleoside, these were the identified degradation products in assays carried out.

Dopaminergic modulation of aldosterone secretion in the rhesus monkey: evidence that dopamine affects the late pathway of aldosterone biosynthesis by inhibiting the conversion of corticosterone to 18-hydroxycorticosterone.

This study was designed to investigate the role of dopaminergic mechanisms in the control of aldosterone secretion. Six rhesus monkeys received metoclopramide (1.25 mg, iv), with 5% dextrose (vehicle) or with dopamine (4 micrograms/kg . min) infusions begun 60 min before the administration of the dopamine antagonists. Metoclopramide, in the presence of vehicle, increased plasma aldosterone concentrations from 4.5 +/- 0.5 ng/dl to a maximum of 26 +/- 4.1 ng/dl and PRL concentrations from 8.1 +/- 1.3 ng/ml to a maximum of 118.4 +/- 7.6 ng/ml. Dopamine infusion inhibited the aldosterone and PRL responses to metoclopramide. The administration of metoclopramide resulted in a rise in plasma 18-hydroxycorticosterone from 10.4 +/- 1.5 ng/dl to a maximum concentration of 41 +/- 4.2 ng/dl. The aldosterone and 18-hydroxycorticosterone responses displayed a parallel time course, with significant responses of both occurring 5 min after metoclopramide administration. Plasma concentrations of electrolytes, PRA, plasma cortisol, 11-deoxycorticosterone, corticosterone, and 18-hydroxy-11-deoxycorticosterone were not altered by metoclopramide. The results of this investigation demonstrate that aldosterone and PRL responses to metoclopramide are mediated by their antagonist activities at dopamine receptors. Rather than simply affecting secretion, dopaminergic mechanisms appear to modulate the late pathway of adrenal glomerulosa biosynthesis of aldosterone. A parallel time course of stimulation of 18-hydroxycorticosterone and aldosterone secretion, with no change in other aldosterone precursors, strongly suggests that dopamine modulates the activity of the glomerulosa 18-hydroxylase enzyme.