Methyl scanning: total synthesis of demethylasterriquinone B1 and derivatives for identification of sites of interaction with and isolation of its receptor(s)

The principle of methyl scanning is proposed for determination of the sites of interaction between biologically active small molecules and their macromolecular target(s). It involves the systematic preparation of a family of methylated derivatives of a compound and their biological testing. As a functional assay, the method can identify the regions of a molecule that are important (and unimportant) for biological activity against even unknown targets, and thus provides an excellent complement to structural biology. Methyl scanning was applied to demethylasterriquinone B1, a small-molecule mimetic of insulin. A new, optimal total synthesis of this natural product was developed that enables the family of methyl scan derivatives to be concisely prepared for evaluation in a cellular assay. The results of this experiment were used to design a biotin-demethylasterriquinone conjugate for use as an affinity reagent. This compound was prepared in tens of milligram quantities in a four-step synthesis.     

CHEMILUMINESCENT REACTIONS OF SO

Abstract— The rapid bimolecular reaction SO + O₃= SO₂+O₂+ 106 kcal/mole
yields electronically excited SO₂ in the ³B ₁ and ¹B₂ states with some vibrational excitation, as well as SO₂ in its electronic ground state. It is shown that k₁ = 1.5 x 10¹² exp (-2100/ RT ) cm² mole^-1 s-¹ and that the formation of electronically excited SO₂ involves higher activation energies.     

A proton NMR study on aggregation of cationic surfactants in water: effects of the structure of the headgroup

 ¹H NMR chemical shifts of solutions of the following cationic surfactants in D₂O were determined as a function of their concentrations: cetyltrimethylammonium chloride, CTACl, a 1 : 1 molar mixture of CTACl and toluene, cetylpyridinium chloride, CPyCl, cetyldimethylphenylam-monium chloride, CDPhACl, cetyldimethylbenzylammonium chloride, CDBzACl, cetyldimethyl-2-phenylethylammonium chloride, CDPhEtACl, and cetyldimethyl-3-phenylpropylammonium chloride, CDPhPrACl. Plots of observed chemical shifts versus [surfactant] are sigmoidal, and were fitted to a model based on the mass-action law. Satisfactory fitting was obtained for the discrete protons of all surfactants. From these fits, we calculated the equilibrium constant for micelle formation, K, the critical micelle concentration, CMC and the chemical shifts of the monomer, δ_mon and the micelle δ_mic. ¹H NMR-based CMC values are in excellent agreement with those which we determined by surface tension measurements of surfactant solutions in H₂O, allowing for the difference in structure between D₂O and H₂O. Values of K increase as a function of increasing the size of the hydrophilic group, but the free energy of transfer per CH₂ group of the phenylalkyl moiety from bulk water to the micellar interface is approximately constant, 1.9±0.1 kJ mol^-1. Values of (δ_mic–δ_mon) for the surfactant groups at the interface, e.g., CH₃–(CH₂)₁₅–N+(CH₃)₂ and within the micellar core, e.g., CH₃–(CH₂)₁₅–N⁺ were used to probe the (average) conformation of the phenyl group in the interfacial region. The picture that emerges is that the aromatic ring is perpendicular to the interface in CDPhACl and is more or less parallel to it in CDBzACl, CDPhEtACl, and CDPhPrACl. Received: 23 February 1996 Accepted: 29 August 1996     

Quantitative determination of N-[trans-2-(dimethylamino)-cyclopentyl]-N-(3',4'-dichlorophenyl)propan amide, its 2H5-labeled analogue and their N-dealkylated metabolites in dog serum by capillary gas chromatography-mass spectrometry

This paper describes the development of a capillary gas chromatographic--mass spectrometric method for the determination of N-[trans-2-(dimethylamino)cyclopentyl]-N-(3',4'-dichlorophenyl)propan amide and its metabolites in serum. The method utilizes an automated sample preparation whereby drug, metabolites and internal standard are extracted from polar serum components by adsorption chromatography onto an XAD-type resin. The N-demethylated metabolites are derivatized by acetylation prior to chromatography. Detection is by mass spectrometry with chemical ionization. This method was utilized to determine levels of unlabeled and pentadeuterated drug and their respective metabolites in canine serum after oral co-administration. No significant kinetic isotope effects were observed for either absorption or metabolism.     

Use of the diffusion gradient thin film method to measure trace metals in fresh waters at low ionic strength

The diffusion gradients in thin films (DGT) method was investigated and used to measure trace metal concentrations in river water. The principle of DGT is that trace metal ions diffuse through a thin polyacrylamide gel film (the diffusion gel layer) and are subsequently immobilised and concentrated on a layer of Chelex-100 resin embedded in another polyacrylamide gel film (the resin gel layer). These layers are mounted in a plastic holder, which exposes a fixed area of the diffusion gel layer to the water being monitored. Replacement of the normal agarose cross-linked diffusion gel with bisacrylamide cross-linked gel altered the ion uptake properties of DGT. The bisacrylamide cross-linked gel weakly, and with little selectivity, bound metal ions prior to their irreversible binding to Chelex-100. Trace metal ion uptake by these DGT devices was thus dependent on ionic strength and temperature, although the ionic strength effect is relatively small for most natural waters and negligible in sea water. The concentrations of Cd, Co, Cu, Ni, Pb, and Zn in the Water of Leith, an urban stream in Dunedin, New Zealand, were measured by DGT and the results compared with total dissolved concentrations of these metals measured in conventional (bottle) samples collected in parallel with the DGT monitoring. Greater than 90% of the total dissolved Cd and Zn; 20-40% of the total dissolved Co, Ni, and Pb; and 5% of the total dissolved Cu was available to the DGT method.     

Evidence for proton pairs in γ-AIOOH (boehmite) from NMR absorption spectra

Continuous-wave and Fourier transform NMR absorption spectra have been recorded for pure γ-AIOOH at room temperature and 130K. The well-resolved doublet lineshape shows that there are proton pairs (intrapair separation $\text{R ? 1.7}$ Å), the presence of which has not previously been recognized. A structural model based on the occupation of two sites consistent with the space group Cmcm is proposed.