Phase dependent encapsulation and release profile of ZIF-based biocomposites.

Biocomposites composed of Zeolitic Imidazolate Frameworks (ZIFs) are generating significant interest due to their facile synthesis, and capacity to protect proteins from harsh environments. Here we systematically varied the composition (i.e. relative amounts of ligand (2-methylimidazole), metal precursor (Zn(OAc)2·2H2O), and protein) and post synthetic treatments (i.e. washes with water or water/ethanol) to prepare a series of protein@ZIF biocomposites. These data were used to construct two ternary phase diagrams that showed the synthesis conditions employed gave rise to five different phases including, for the first time, biocomposites based on ZIF-CO3-1. We examined the influence of the different phases on two properties relevant to drug delivery applications: encapsulation efficiency and release profile. The encapsulation efficiencies of bovine serum albumin and insulin were phase dependent and ranged from 75% to 100%. In addition, release profiles showed that 100% protein release varied between 40 and 300 minutes depending on the phase. This study provides a detailed compositional map for the targeted preparation of ZIF-based biocomposites of specific phases and a tool for the straightforward analysis of the crystalline phases of ZIF based materials (web application named "ZIF phase analysis"). These data will facilitate the progress of ZIF bio-composites in the fields of biomedicine and biotechnology.

Novel pyrazole compounds for pharmacological discrimination between receptor-operated and store-operated Ca(2+) entry pathways.

BACKGROUND AND PURPOSE: Pyrazole derivatives have recently been suggested as selective blockers of transient receptor potential cation (TRPC) channels but their ability to distinguish between the TRPC and Orai pore complexes is ill-defined. This study was designed to characterize a series of pyrazole derivatives in terms of TRPC/Orai selectivity and to delineate consequences of selective suppression of these pathways for mast cell activation. EXPERIMENTAL APPROACH: Pyrazoles were generated by microwave-assisted synthesis and tested for effects on Ca(2+) entry by Fura-2 imaging and membrane currents by patch-clamp recording. Experiments were performed in HEK293 cells overexpressing TRPC3 and in RBL-2H3 mast cells, which express classical store-operated Ca(2+) entry mediated by Orai channels. The consequences of inhibitory effects on Ca(2+) signalling in RBL-2H3 cells were investigated at the level of both degranulation and nuclear factor of activated T-cells activation. KEY RESULTS: Pyr3, a previously suggested selective inhibitor of TRPC3, inhibited Orai1- and TRPC3-mediated Ca(2+) entry and currents as well as mast cell activation with similar potency. By contrast, Pyr6 exhibited a 37-fold higher potency to inhibit Orai1-mediated Ca(2+) entry as compared with TRPC3-mediated Ca(2+) entry and potently suppressed mast cell activation. The novel pyrazole Pyr10 displayed substantial selectivity for TRPC3-mediated responses (18-fold) and the selective block of TRPC3 channels by Pyr10 barely affected mast cell activation. CONCLUSIONS AND IMPLICATIONS: The pyrazole derivatives Pyr6 and Pyr10 are able to distinguish between TRPC and Orai-mediated Ca(2+) entry and may serve as useful tools for the analysis of cellular functions of the underlying Ca(2+) channels.

Automated library generation using sequential microwave-assisted chemistry. Application toward the Biginelli multicomponent condensation.

The concept of automated sequential microwave-assisted library synthesis is introduced. For this purpose a dedicated single-mode microwave reactor with a robotics interface including a liquid handler and gripper was employed. The liquid handler allows dispensing of reagents into the Teflon sealed reaction vials, while the gripper moves each sealed vial in and out of the microwave cavity after irradiation. This technology was employed for the Biginelli three-component cyclocondensation reaction. A diverse set of 17 CH-acidic-carbonyl compounds (1A-Q), 25 aldehydes (2a-y), and 8 urea/thioureas (3alpha-varphi) was used in the preparation of a dihydropyrimidine (DHPM) library. Out of the full set of 3400 possible DHPM derivatives, a representative subset of 48 analogues was prepared using automated addition of building blocks and subsequent sequential microwave irradiation of each process vial. For most building block combinations 10 min of microwave flash heating at 120 degrees C using AcOH/EtOH (3:1) and 10 mol % Yb(OTf)(3) as solvent/catalyst system proved to be successful, leading to an average isolated yield of 52% of DHPMs with >90% purity. For some building block combinations the general conditions were modified, for example, by changing the solvent, catalyst, reaction temperature, or irradiation time. This flexibility is a distinct advantage of sequential over parallel microwave-assisted processes where all reactions are exposed to the same irradiation conditions. When the unattended automation capabilities of the microwave synthesizer are used, a library of this size can be synthesized within 12 h.

Absolute configuration in 4-alkyl- and 4-aryl-3,4-dihydro-2(1H)-pyrimidones: a combined theoretical and experimental investigation.

Structural features (orientation of the carboxyl group, ring puckering), electronic absorption, and circular dichroism spectra of 4-alkyl- and 4-aryl-dihydropyrimidones 1-5 are calculated by semiempirical (AM1, INDO/S), ab initio (HF/6-31G, CIS/6-31G, RPA/6-31G), and density functional theory (B3LYP/6-31G) methods. These calculations allow an assignment of the absolute configuration by comparison of simulated and experimental CD spectra. Although the ab initio methods greatly overestimate electronic transition energies, the general appearance of the experimental CD spectra is quite nicely reproduced by these calculations. Thus, comparison of experimental with calculated CD spectra is a reliable tool for the assignment of the absolute configuration. For 4-methyl derivatives 1, the first enantiopure DHPM examples with no additional aromatic substituent, the stereochemistry at C4 provided by the theoretical results is confirmed by X-ray structure determination of the diastereomeric salt 6. Additional support is the consistent HPLC elution order found for all investigated DHPMs on a cellulose-derived chiral stationary phase.

Chiral separation of pharmacologically active dihydropyrimidinones with carboxymethyl-beta-cyclodextrin.

We report on the chiral separation of pharmacologically active dihydropyrimidinones by capillary electrophoresis (CE) using carboxymethyl-beta-cyclodextrin as chiral selector. The influence of selector concentration, pH, and the addition of varying amounts of methanol is investigated. Out of 21 compounds investigated, 19 were resolved, 13 with baseline separation.

Iminopropadienethiones, Ar=N=C=C=C=S.

Aryliminopropadienethiones 9 have been generated by flash vacuum thermolysis of isoxazolones of the type 5 and characterized by mass spectrometry and matrix isolation IR spectroscopy in conjunction with DFT calculations and chemical trapping.

Recent advances in the Biginelli dihydropyrimidine synthesis. New tricks from an old dog.

In 1893, P. Biginelli reported the synthesis of functionalized 3, 4-dihydropyrimidin-2(1H)-ones (DHPMs) via three-component condensation reaction of an aromatic aldehyde, urea, and ethyl acetoacetate. In the past decade, this long-neglected multicomponent reaction has experienced a remarkable revival, mainly due to the interesting pharmacological properties associated with this dihydropyrimidine scaffold. In this Account, we highlight recent developments in the Biginelli reaction in areas such as solid-phase synthesis, combinatorial chemistry, and natural product synthesis.

Highly versatile solid phase synthesis of biofunctional 4-aryl-3,4-dihydropyrimidines using resin-bound isothiourea building blocks and multidirectional resin cleavage.

A series of pharmacologically active, functionalized 4-aryl-3,4-dihydropyrimidine-5-carboxylates (DHPMs) are prepared by a versatile novel solid phase approach. In the key step, a polymer-bound thiouronium salt is condensed with unsaturated 1-ketoesters. The resulting polymer bound 1,4-dihydropyrimidines are cleaved from the resin employing multidirectional resin cleavage strategies.

Biologically active dihydropyrimidones of the Biginelli-type--a literature survey.

In 1893, the synthesis of functionalized 3,4-dihydropyrimidin-2(1H)-ones (DHPMs) via three-component condensation reaction of an aromatic aldehyde, urea and ethyl acetoacetate was reported for the first time by P. Biginelli. In the past decades, such Biginelli-type dihydropyrimidones have received a considerable amount of attention due to the interesting pharmacological properties associated with this heterocyclic scaffold. In this review, we highlight recent developments in this area, with a focus on the DHPMs recently developed as calcium channel modulators, alpha(1a) adrenoceptor-selective antagonists and compounds that target the mitotic machinery.

Determination of absolute configuration in 4-aryl-3, 4-dihydro-2(1H)-pyrimidones by high performance liquid chromatography and CD spectroscopy.

The absolute configuration of three 4-aryl-3, 4-dihydro-2(1H)-pyrimidones (Biginelli compounds, DHPMs) was established by comparison of the typical circular dichroism (CD) spectra of individual enantiomers with reference samples of known absolute configuration. The enantiomers were obtained by semipreparative separation of racemic mixtures on a Chiralcel OD-H chiral stationary phase. The method was used to establish the enantiopreference of various lipases in biocatalytic kinetic resolution experiments employing activated DHPM esters.

Cross-conjugated and pseudo-cross-conjugated mesomeric betaines, XVIII: Bicyclic mesoionic pyrimidines with cardiovascular activity.

Reaction of alpha-Anilino-azines 1a-i with activated malonates (magic malonates) 2a-e leads to bicyclic mesoionic systems 3-7. Out of these, pyrimido[1,2-alpha]pyrimidines 3b,d are active as cardiotonics, whereas pyrimido[1,2-alpha]pyrimidines 4a-g show significant anti-anginal and anti-hypertensive activity.