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D J Abraham

Publications and source records attributed to D J Abraham.

At least 91 records · Page 5Linked to original sources
PubMed1990

Comparison of crystal and solution hemoglobin binding of selected antigelling agents and allosteric modifiers.

This paper details comprehensive binding studies (solution and X-ray) of human hemoglobin A with a group of halogenated carboxylic acids that were investigated as potential antisickling agents. It is, to our knowledge, the first study to compare solution and crystal binding for a series of compounds under similar high-salt conditions used for cocrystallization. The compounds include [(3,4-dichlorobenzyl)oxy]acetic acid, [(p-bromobenzyl)oxy]acetic acid, clofibric acid, and bezafibrate. The location and stereochemistry of binding sites have been established by X-ray crystallography, while the number of binding sites and affinity constants were measured by using equilibrium dialysis. The solution binding studies were conducted with deoxygenated hemoglobin and carbonmonoxyhemoglobin with low (50 mM) and high (2 M) salt concentrations. It was concluded that the observed crystal structures are consistent with the binding observed in solution and that the number of binding sites is independent of salt concentration, while the binding constant increases with increasing salt concentration. The studies also reveal that relatively small changes in the chemical structure of a drug molecule can result in entirely different binding sites on the protein. Moreover, the X-ray studies provide a possible explanation for the multiplicity in function exhibited by these compounds as allosteric modulators and/or antisickling agents. Finally, the studies that these compounds bind differently to the R and T states of hemoglobin, an observation of special significance to the original design of these agents.

Allosteric Regulation↗
PubMed1990

Stereochemical characterization of the diastereomers of the amobarbital N-glucosides excreted in human urine.

The stereochemistry associated with the amobarbital N-glucoside diastereomers (1a and 1b) that are excreted by humans in urine is unknown. Using X-ray crystallography, the absolute configuration of 1b was determined to be S (C-5 position of the barbiturate ring). Following oral administration of amobarbital to Caucasians and Orientals, from 5 to 25% of the dose of amobarbital was excreted in the urine as 1b. The other diastereomer, 1a, accounted for less than 0.1 to 0.2% of the dose in four individuals, with none detected in nine individuals. The rate constants, kf,1b, determined from the urinary excretion of 1b were lower than those previously reported for unresolved amobarbital N-glucosides. However, based on the urinary excretion of 1b, the rate constants, K, for elimination of amobarbital in Caucasians and Orientals were similar to those previously determined from the serum levels of amobarbital and the urinary excretion of unresolved amobarbital N-glucosides. In previous studies of the N-glucosylation of amobarbital, it is likely that a single N-glucose diastereomer, 1b, was being observed.

Adult↗
PubMed1989

Design, synthesis, and testing of potential antisickling agents. 7. Ethacrynic acid analogues.

In search of a drug to treat sickle cell anemia, several analogues of the diuretic ethacrynic acid (ECA) have been synthesized and found equivalent in antigelling potency to ECA, but they have moderate or little diuretic activity. Structure-activity studies revealed that most of the highly active derivatives contain an acryloyl moiety. The latter functionality reacts covalently with protein sulfhydryl groups via a Michael addition reaction. Other derivatives, which lack the acryloyl moiety, showed notably lower antigelling activity. Since the antigelling assay is run under anaerobic conditions, activity implies a stereochemical inhibition of polymerization of deoxyhemoglobin S. The solubility ratios obtained from [HbS drug]/[HbS control] of several compounds (Table I) are near those expected for a drug with clinical potential (1.06-1.20 at tolerable doses in vivo).

Animals↗
PubMed1989

Quantitative evaluation of congo red binding to amyloid-like proteins with a beta-pleated sheet conformation.

The binding of Congo red to several purified amyloid-like peptides having a beta-pleated sheet conformation was quantitatively examined. Congo red binds preferentially to the beta-pleated sheet conformation of both insulin fibrils and poly-L-lysine. Congo red does not bind nearly so well to poly-L-serine or polyglycine, despite the fact that these peptides also have a beta-pleated sheet conformation. Binding to insulin fibrils was saturable with an apparent Bmax of 2 moles of Congo red per mole of insulin fibrils and an apparent KD of 1.75 x 10(-7) M. Binding to beta-poly-L-lysine was similar but had a much higher apparent Bmax of 43. Binding of Congo red to beta-poly-L-lysine was pH dependent and appeared to be determined by the number of protonated lysine residues in the 250 amino acid peptide. We present a new hypothesis in which Congo red binds to amyloid-like proteins via bonds between the two negatively charged sulfonic acid groups of Congo red and two positively charged amino acid residues of two separate protein molecules which are properly oriented by virtue of the beta-pleated sheet conformation of the peptide backbone.

Amyloid↗
PubMed1989

Two simple methods for quantifying low-affinity dye-substrate binding.

Binding with low-affinity ligands, such as histological dyes, can be difficult to quantitate owing to the dissociation of bound ligand with washing or the retention of nonspecifically bound ligand because of incomplete washing. The present report describes two simple, rapid methods of discriminating bound from free ligand without the need for washing steps. One method is based on the spectral changes induced in a dye ligand, Congo red, on binding to the "receptor" insulin fibrils. This method discriminates spectrophotometrically between bound and free ligand without requiring any physical separation of the two forms. No radioactive ligands are necessary, and, by using disposable cuvettes, the entire binding assay can be done in a single container without the need for transfers. The second method employs a non-traditional filtration approach that avoids the need for a washing step by measuring the decrease in concentration of the dye ligand in the filtrate rather than by applying the usual approach of measuring the absolute amount of ligand bound to the precipitated "receptor." Both methods show saturation of binding sites and give similar values for the KD and Bmax.

Congo Red↗
PubMed1988

Evaluation of the side arm of (naphthylvinyl)pyridinium inhibitors of choline acetyltransferase.

A number of quaternary salts of trans-4-(beta-1-naphthylvinyl)pyridine (NVP) were synthesized and evaluated as inhibitors of the enzymes choline acetyltransferase (ChAT) and acetylcholinesterase (AChE). Structural variations in the side arm attached to the pyridine nitrogen atom demonstrate that an inductive effect is small but significant for activity. Inhibition of ChAT by alkylated derivatives decreases when electron-withdrawing groups are placed in the side chain. Substitution of a methyl group on the pyridine ring only slightly affects activities toward ChAT and AChE. When the pyridinium moiety is replaced by an imidazolium ring, no ChAT inhibition was observed. The imidazolium compound, however, was a weak inhibitor of AChE. For design of affinity columns for purification of ChAT, the data also supports the use of long chain alkylated amide derivatives of NVP.

Animals↗
PubMed1988

Filamentous proteins in Alzheimer's disease: new insights through molecular biology.

Recent advances in the study of the neurofibrillary tangles (NFT), neuritic plaques (NP), and cerebrovascular amyloid found in the brain of Alzheimer's disease (AD) victims have greatly expanded our understanding of the molecular biology of this disease. Paired helical filaments (PHF) are the primary intracellular filamentous deposit. They appear to be distinct from normal cytoskeletal proteins, but they do contain a significant component of a microtubule-associated protein called 'tau'. Amyloid fibrils make up the extracellular filamentous deposits in AD. Amyloid fibrils are composed of a small protein of about 43 amino acids which has been sequenced. Some investigators suggest PHF are composed of this same protein, but this remains debatable. Molecular genetic studies have shown that a gene which codes for a larger protein containing the sequence of the 'beta-amyloid protein' exists in many tissues and in many species. Interestingly, this gene is located on chromosome 21 which is also the location of the familial AD gene, but these 2 genes are distinct. Several hypotheses exist on the origin of these abnormal deposits and range from neuronal origin to synthesis outside of the CNS with transportation via the blood. The implications of these recent advances are great and include the possibility of accurate and early antemortem diagnostic tests for AD, as well as therapeutic manipulation of the synthesis, deposition, or removal of these filamentous proteins.

Alzheimer Disease↗
PubMed1987

Extension of the fragment method to calculate amino acid zwitterion and side chain partition coefficients.

The fragment method of calculating partition coefficients (P) has been extended to include the common amino acids (AAs). The results indicate that polar and charged side chains influence the hydrophobicity of atoms in the side chain in a predictable manner. Field effects, as evidenced through polar proximity factors and bond factors, need to be considered for accurate estimation of transfer phenomena. The calculated log P and delta G degree ' values of the 20 AAs agree well with the observed values. Pro calculates to be more hydrophilic than the observed log P. Hydrophobicity scales for peptide side chain residues are compared and evaluated in terms of suitability. Calculated pi values for nonpolar side chain residues agree well with the observed values; calculated values for uncharged polar side chain residues deviate by about 0.6 log units except for Gln and Cys; and polar side chain residues with charged side chains calculate as too hydrophilic. Reasons for the differences are explored. We also suggest that tightly bound water to polar moieties in amino acids and peptides may be transferred into the octanol phase during partitioning experiments. A quantitative methodology is presented which characterizes the thermodynamic partitioning of groups and individual atoms in amino acids and proteins.

Amino Acids↗
PubMed1987

Design, synthesis, and testing of potential antisickling agents. 10. (2,2-Dimethylchroman-6-yl)alkanoic acids.

Five (2,2-dimethylchroman-6-yl)alkanoic acids were synthesized and tested for antigelling activities. It was envisioned that these agents might bind via hydrophobic bonding to nonpolar sites of the "donor-acceptor" regions of hemoglobin S. Several (2,2-dimethylchroman-6-yl)alkanoic acids containing 1-4 carbon atoms on the side-chain residue were designed to interact at the acceptor site, were synthesized, and were found to be moderately potent antigelling agents. The weak activity observed for two of the acids at low concentrations is rationalized in terms of weak binding affinities or multiple binding to active and nonactive sites. The effect of these compounds on shifting the allosteric equilibrium was small or negligible. The low toxicity of one of the (2,2-dimethylchroman-6-yl)alkanoic acids demonstrates the potential use of yet another class of compounds that can be modified in the development of antisickling agents.

Antisickling Agents↗
PubMed1987

Design, synthesis, and testing of potential antisickling agents. 9. Cyclic tetrapeptide homologs as mimics of the mutation site of hemoglobin S.

As part of our continuing search for new agents which might be useful for the treatment of sickle-cell anemia, we have synthesized two cyclic tetrapeptide homologs, cyclo(-Val-Glu[-Thr-Pro-]-OH) (1a) and cyclo(-Phe-Glu[-Thr-Pro-]-OH (1b), and a tetrapeptide lactone homolog cyclo(H-Thr-Pro-Val-Glu-OH) (2). The intent was that these peptides would mimic a tetrapeptide region around the mutation site of HbS and thus be able to bind at the acceptor site of HbS and thereby inhibit polymerization. The synthesis of the linear peptides was accomplished in solution using both the polymeric reagent (PHBT) and DCC/HOBT methods; cyclization was accomplished by an improved method. 13C-n.m.r. studies were performed which allowed us to assign the conformation about the Thr-Pro bond in 1a and 2 as trans. The cyclic peptides were tested for their ability to increase the solubility of HbS under deoxygenating conditions, but only 1a had any antigelling activity, albeit low.

Antisickling Agents↗
PubMed1987

Itraconazole therapy for fungal keratitis.

110 cases of keratomycosis were treated with itraconazole administered either orally or topically or by both routes. Excellent or moderate response was obtained in 69 percent of the patients. Ulcers due to Aspergillus and dematiaceous fungi and non-sevee ulcers responded better than did severe ulcers and ulcers due to Fusarium. A combination of oral and topical itraconazole yielded the highest percentage of excellent or moderate responses.

Adolescent↗
PubMed1987

Design, synthesis, and testing of potential antisickling agents. 6. Rheologic studies with active phenoxy and benzyloxy acids.

In vitro rheologic studies have been performed with homozygous sickle erythrocytes in the presence of phenoxy and benzyloxy antigelling agents. The results correlate with those obtained previously from a standard hemoglobin S solubility assay (C-sat) that monitors the inhibition of hemoglobin S polymerization in solution. The order of activity of compounds in both studies was found to be about the same. The rheologic assay monitors the reduction in pressure (130 mm Hg sickled vs 9 mm Hg nonsickled) needed to push erythrocytes through a 5 micron filter. Among the drugs studied, the antilipidemic drug Clofibrate reduced the filtration pressure by as little as 9%, and a derivative of the diuretic agent ethacrynic acid (Merck-595, 595) reduced it by as much as 94%. Human Serum Albumin (HSA) (0.1% wt/vol) drastically reduced the activity of the noncovalently-acting aromatic acids by requiring at least three times the concentration of drug to obtain similar reductions in pressure. Two covalently-acting compounds, ethacrynic acid and Merck-595, 595, strongly increased the filterability of homozygous S erythrocytes at 1 and 3 mM drug concentrations even in the presence of HSA. No reason is given for the strong inhibitory effect of HSA at such high drug to HSA concentrations.

Anemia, Sickle Cell↗
PubMed1986

Solubility properties in polymers and biological media: 6. An equation for correlation and prediction of solubilities of liquid organic nonelectrolytes in blood.

Gas/blood partition coefficients, Kgb, and molar solubilities in blood, Sb, of 27 liquid aliphatic nonelectrolyte solutes are described by the equation: log Sb congruent to log (Sg/Kgb) = 1.35-3.05V/100-0.28 pi* + 3.58 beta (at 37 degrees C) r = 0.9870 and SD = 0.144 where Sg is the solute concentration in its saturated vapor at 37 degrees C, given by Sg = Patm/25.4; V is the solute molar volume; and pi* and beta are parameters that are measures of solute dipolarity/polarizability and hydrogen bond acceptor basicity, respectively. The above equation applies to compounds with calculated log solubilities greater than -1.5. A nondipolar solubility reservoir provided by hydrophobic pockets in hemoglobin leads to greater than predicted solubilities for the less soluble alkanes, alkylbenzenes, and chlorobenzenes. It is suggested that equations similar to the one above can be constructed to describe and predict gas-lipid and gas-homogenized tissue, and hence blood-lipid, blood-tissue, tissue-lipid, and tissue-tissue partition coefficients of pharmacologically and toxicologically active solutes.

Hemoglobins↗
PubMed1986

Neurochemistry of aging. 1. Toxins for an animal model of Alzheimer's disease.

A chronic deficiency in central cholinergic function has been implicated in a number of neuropsychiatric diseases including Alzheimer's disease. Until recently, animal models that simulate the neurochemical conditions that appear to cause these diseases in humans, as a result of a direct manipulation of the central cholinergic system, were not available. Over the past few years, however, we have been successful in developing a cholinotoxin, 1-ethyl-1-(2-hydroxyethyl)aziridinium chloride (AF64A), which has the potential to serve as a novel compound in developing animal models of human brain disorders in which a cholinergic hypofunction has been implicated. In this paper are described the design, synthesis, and testing of several structural analogues of AF64A as potential cholinotoxins, by evaluating them for their ability to inhibit high-affinity choline transport and their affinity toward brain muscarinic receptors. One of the compounds, 1-cyclopropyl-1-(2-hydroxyethyl)aziridinium chloride (i.e. aziridine analogue of 13) was found to have a remarkably high affinity (about 40 times higher than AF64A) toward brain muscarinic receptors.

Alzheimer Disease↗