ABSTRACT
A series of carboxylic acids were prepared from a propargylglycine scaffold and tested for efficacy as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for four enzymes within the MMP family. The inhibitors were typically potent against collagenase-3 (MMP-13) and gelatinase A (MMP-2), while they spared collagenase-1 (MMP-1) and only moderately inhibited stromelysin (MMP-3). Compound 40 represents a typical inhibition profile of a compound with reasonable potency. Introduction of polar groups was required in order to generate inhibitors with acceptable water solubility, and this often resulted in a loss of potency as in compound 63. High serum protein binding proved to be a difficult hurdle with many compounds such as 48 showing >99% binding. Some compounds such as 64 displayed approximately 90% binding, but no reliable method was discovered for designing molecules with low protein binding. Finally, selected data regarding the pharmacokinetic behavior of these compounds is presented.
Subject(s)
Alkynes/chemical synthesis , Carboxylic Acids/chemical synthesis , Glycine/analogs & derivatives , Glycine/chemical synthesis , Metalloendopeptidases/antagonists & inhibitors , Protease Inhibitors/chemical synthesis , Alkynes/chemistry , Carboxylic Acids/chemistry , Glycine/chemistry , Humans , Matrix Metalloproteinase 3/chemistry , Matrix Metalloproteinase Inhibitors , Metalloendopeptidases/chemistry , Models, Molecular , Protease Inhibitors/chemistry , Structure-Activity RelationshipABSTRACT
Bioanalytical methods based on automated solid-phase extraction (SPE) and high-performance liquid chromatography with electrospray tandem mass spectrometry (LC-MS-MS) have been developed and utilized for the determination of MMP inhibitors in plasma and cartilage tissues. The SPE methods were automated using a 96-well extraction plate and a 96-channel programmable liquid-handling workstation. The LC-MS-MS methods were developed using a rapid gradient LC separation, followed by sample introduction through an ionspray interface in the positive ion mode and tandem mass spectrometric detection with selected reaction monitoring. In the optimized SPE methods, crude plasma or ground cartilage supernatant samples were loaded onto an SPE plate to remove proteins and other interfering components in the matrixes to render relatively clean extracts for LC-MS-MS analysis. Compared to the simple plasma protein precipitation method, the automated SPE method afforded significant time-saving in sample preparation and improved sensitivity in MS detection. The methods were validated and successfully applied to the analysis of protease inhibitors in plasma and cartilage tissues.
Subject(s)
Cartilage/chemistry , Protease Inhibitors/analysis , Animals , Autoanalysis , Chromatography, High Pressure Liquid , Mass Spectrometry , Protease Inhibitors/blood , RatsABSTRACT
A series of hydroxamates was prepared from an aminoproline scaffold and tested for efficacy as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for five enzymes within the MMP family, and a number of inhibitors, such as compound 47, display broad-spectrum activity with sub-nanomolar potency for some enzymes. Modifications of the P1' portion of the molecule played a key role in affecting both potency and selectivity within the MMP family. Longer-chain aliphatic substituents in this region of the molecule tended to increase potency for MMP-3 and decrease potency for MMP-1, as exemplified by compounds 48-50, while aromatic substituents, as in compound 52, generated broad-spectrum inhibition. The data is rationalized based upon X-ray crystal data which is also presented. While the in vitro peroral absorption seemed to be less predictable, it tended to decrease with longer and more hydrophilic substituents. Finally, a rat model of osteoarthritis was used to evaluate the efficacy of these compounds, and a direct link was established between their pharmacokinetics and their in vivo efficacy.
Subject(s)
Hydroxamic Acids/chemical synthesis , Metalloendopeptidases/antagonists & inhibitors , Proline/analogs & derivatives , Proline/chemical synthesis , Protease Inhibitors/chemical synthesis , Animals , Cartilage, Articular/pathology , Crystallography, X-Ray , Humans , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Iodoacetates , Male , Matrix Metalloproteinase 3/chemistry , Models, Molecular , Osteoarthritis, Knee/chemically induced , Osteoarthritis, Knee/drug therapy , Osteoarthritis, Knee/pathology , Proline/chemistry , Proline/pharmacology , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Rats , Rats, Sprague-Dawley , Structure-Activity RelationshipABSTRACT
Diethylene glycol monobutyl ether (DGBE) is a solvent used in some liquid hard surface cleaners. We evaluated the inhalation component of consumer exposure in the home to DGBE from the use of cleaning products containing up to 9% DGBE. Several experiments were conducted with restricted room air flow, exaggerated amounts of cleaning solutions, and no rinsing in order to develop an exposure scenario that would exceed exposures likely encountered by consumers. DGBE vapors in the air were monitored by collection on charcoal tubes, followed by desorption and quantitation by gas chromatography. Air was collected from the centre of the room and from the breathing zone of the person doing the washing task. Room air concentrations of DGBE showed peak values between one and three hours after task initiation; DGBE concentrations then gradually decreased with time. Peak concentrations did not exceed 1.6 ppmv. The total DGBE in the air at the time of maximum air concentrations accounted for only 1 to 3% of the DGBE on the washed surfaces. The person doing the washing task was exposed to average DGBE concentrations in the breathing zone below 0.8 ppmv in all experiments. The methods described for measuring DGBE concentrations in air are generally applicable to other solvents and easily adaptable to various experimental situations.