Synthesis and kinetic studies of an amidine-containing phosphonofluoridate: a novel potent inhibitor of trypsin-like enzymes.

The amidine-containing alpha-aminoalkyl phosphonofluoridate 3 (Cbz-(4-AmPhGly)P(OPh)(F)) is a very potent inhibitor of trypsin-like enzymes. It was prepared by hydrolyzing the corresponding phosphonate diphenyl ester 4 followed by reaction of fluoride with the phosphonochloridate prepared from the intermediate phosphonic acid monoester 5. Compound 3 is the most potent amidine-containing organophosphorus inhibitor yet reported for trypsin-like enzymes. It inhibits trypsin and thrombin with second-order rate constants (Kobs/[I]) of 2.6 x 10(5) M-1 s-1 and 1.0 x 10(5) M-1 s-1, respectively, showing a 130-fold and a 1250-fold rate enhancement over the corresponding diphenyl ester (4). It also inactivates trypsin 2 orders of magnitude more potently than simple phosphonofluoridates such as DFP,1 Sarin and Soman. The phosphonofluoridate 3 does not inhibit other serine proteases such as porcine pancreatic elastase (PPE) and the esterase acetylcholinesterase (AChE). The phosphonofluoridate 3 is hydrolyzed rapidly in buffer solution and has a t1/2 of 4.5 s at pH 7.5.

Synthesis and evaluation of diphenyl phosphonate esters as inhibitors of the trypsin-like granzymes A and K and mast cell tryptase.

Thirty-six new amino acid and peptidyl diphenyl phosphonate esters were synthesized and evaluated to identify potent and selective inhibitors for four trypsin-like proteases: lymphocyte granzymes A and K, human mast cell tryptase, and pancreatic trypsin. Among five Cbz derivatives of Lys and Arg homologues, Z-(4-AmPhe)P(OPh)2 is the most potent inhibitor for granzyme A, and Z-LysP(OPh)2 is the best inhibitor for granzyme K, mast tryptase, and trypsin. The amidino P1 residue D,L-(4-AmPhGly)P(OPh)2 was utilized in a series of compounds with several different N-protecting groups and systematic substitutions at P2 in Cbz-AA derivatives and at P3 in Cbz-AA-Ala derivatives. Generally, these phosphonates inhibit granzyme A and trypsin more potently than granzyme K and tryptase. The P2 Thr and Ala dipeptide phosphonates, Cbz-AA-(4-AmPhGly)P(OPh)2, are the most potent inhibitors for granzyme A, and Cbz-Thr-(4-AmPhGly)P(OPh)2 (kobs/[I] = 2220 M-1 s-1) was quite specific with much lower inhibition rates for granzyme K and trypsin (kobs/[I] = 3 and 97 M-1 s-1, respectively) and no inhibition with tryptase. The most effective inhibitor of granzyme A was Ph-SO2-Gly-Pro-(4-AmPhGly)P(OPh)2 with a second-order rate constant of 3650 M-1 s-1. The most potent inhibitor for granzyme K was 3, 3-diphenylpropanoyl-Pro-(4-AmPhGly)P(OPh)2 with a kobs/[I] = 1830 M-1 s-1; all other phosphonates inhibited granzyme K weakly (kobs/[I] < 60 M-1 s-1). Human mast cell tryptase was inhibited slowly by these phosphonates with Cbz-LysP(OPh)2 as the best inhibitor (kobs/[I] = 89 M-1 s-1). The overall results suggest that scaffolds of Phe-Thr-(4-AmPhe) and Phe-Pro-Lys will be useful to create selective phosphonate inhibitors for granzymes A and K, respectively, and that P4 substituents offer opportunities to further enhance selectivity and reactivity.

Parallel Algorithms for Image Template Matching on Hypercube SIMD Computers.

This correspondence presents several parallel algorithms for image template matching on an SIMD array processor with a hypercube interconnection network. For an N by N image and an M by M window, the time complexity is reduced from O(N2M2) for the serial algorithm to O(M2/K2 + M * log2 N/K + log2 N * log2 K) for the N2K2-PE system (1 ¿ K ¿ M), or to O(N2M2/L2) for the L2-PE system (L ¿ N). With efficient use of the inter-PE communication network, each PE requires only a small local memory, many unnecessary data transmissions are eliminated, and the time complexity is greatly reduced.

A VLSI Systolic Architecture for Pattern Clustering.

Cluster analysis is a valuable tool in exploratory pattern analysis, especially when very little prior information about the data is available. In unsupervised pattern recognition and image segmentation applications, clustering techniques play an important role. The squared-error clustering technique is the most popular one among different clustering techniques. Due to the iterative nature of the squared-error clustering, it demands substantial CPU time, even for modest numbers of patterns. Recent advances in VLSI microelectronic technology triggered the idea of implementing the squared-error clustering directly in hardware. A two-level pipelined systolic pattern clustering array is proposed in this paper. The memory storage and access schemes are designed to enable a rhythmic data flow between processing units. Each processing unit is pipelined to further enhance the system performance. The total processing time for each pass of pattern labeling and cluster center updating is essentially dominated by the time required to fetch the pattern matrix once. Detailed architectural configuration, system performance evaluation, and simulation experiments are presented. The modularity and the regularity of the system architecture make it suited for VLSI implementations.