Miriam N. Jacobs

Compound lipophilicity for substrate binding to human P450s in drug metabolism

Compound lipophilicity is of key importance to P450 binding affinity and enzyme selectivity. Here, lipophilicity is discussed with reference to the human drug-metabolizing P450 enzymes of families CYP1, CYP2 and CYP3. From an extensive compilation of log P values for P450 substrates, and by analysis of relationships between partitioning energy and substrate-binding free energy, the relevance of lipophilicity and other factors pertaining to P450 binding affinity is explained, leading to the formulation of lipophilicity relationships within substrates of each human P450 enzyme involved in drug metabolism. Furthermore, log P values for P450 substrates appear to represent markers for enzyme selectivity. Together with the important roles of hydrogen bonding and pi-pi stacking interaction energies, the desolvation of the P450 active site makes a major contribution to the overall substrate-binding energy and, consequently, a good agreement with experimental information is reported based on this analysis.

Steroid hormone receptors and dietary ligands: a selected review.

Members of the nuclear steroid hormone superfamily mediate essential physiological functions. Steroid hormone receptors (SHR) act directly on DNA, regulate the synthesis of their target genes and are usually activated by ligand binding. Both endogenous and exogenous compounds and their metabolites may act as activators of SHR and disruptors of endocrine, cellular and lipid homeostasis. The endogenous ligands are generally steroids such as 17beta-oestradiol, androgens, progesterone and pregnenolone. The exogenous compounds are usually delivered through the diet and include non-steroidal ligands. Examples of such ligands include isoflavanoids or phytooestrogens, and food contaminants such as exogenous oestrogens from hormone-treated cattle, pesticides, polychlorinated biphenyls and plasticisers. Certain drugs are also ligands; so nuclear receptors are also important drug targets for intervention in disease processes. The present review summarises recent reports on ligand-activated SHR that describe the selective regulation of a tightly-controlled cross-talking network involving exchange of ligands, and the control of major classes of cytochrome P450 (CYP) isoforms which metabolise many bioactive exogenous compounds. Many CYP have broad substrate activity and appear to be integrated into a coordinated metabolic pathway, such that whilst some receptors are ligand specific, other sensors may have a broader specificity and low ligand affinity to monitor aggregate levels of inducers. They can then trigger production of metabolising enzymes to defend against possible toxic nutrients and xenobiotic compounds. The influence of dietary intakes of nutrients and non-nutrients on the human oestrogen receptors (alpha and beta), the aryl hydrocarbon receptor, the pregnane X receptor, the constitutive androstane receptor, and the peroxisome proliferator-activated receptors (alpha and gamma), can be examined by utilising computer-generated molecular models of the ligand-receptor interaction, based on information generated from crystallographic data and sequence homology. In relation to experimental and observed data, molecular modelling can provide a scientifically sound perspective on the potential risk and benefits to human health from dietary exposure to hormone-mimicking chemicals, providing a useful tool in drug development and in a situation of considerable public concern.

Investigation of polychlorinated dibenzo-p-dioxins, dibenzo-p-furans and selected coplanar biphenyls in Scottish farmed Atlantic salmon (Salmo salar).

Farmed and wild Scottish Atlantic salmon were obtained from retail suppliers, producers, and Stirling University in Scotland during January, 1999, for determination of 17 2,3,7,8-C1-substituted PCDDs and PCDFs, and seven nonortho- and mono-ortho-PCBs. The study confirms previous reports of relatively high concentrations of PCDDs, PCDFs and, especially, PCBs in farmed Scottish salmon. The results indicate that high consumption of salmon, particularly by children under 5 years, could lead to intakes above the tolerable daily intake (TDI) and tolerable weekly intake (TWI) for these chemicals, especially the PCBs, when combined with mean or high level intakes from the typical UK diet. These results suggest further investigation of farmed salmon and salmon feed, including feed fortified with fish oil and feed fortified with selected vegetable oils, is warranted.

Human carcinogens: an evaluation study via the COMPACT and HazardExpert procedures

The results of computer-optimized molecular parametric analysis of chemical toxicity (COMPACT) and Hazard Expert evaluations on 14 established human carcinogens are reported. The concordances between COMPACT and carcinogenicity (71%%) and between HazardExpert and carcinogenicity (57%%) are significantly improved when taken in combination, where all 14 carcinogens are correctly identified by the two systems used in conjunction. However, if a negative energy of the highest occupied molecular orbital (EHOMO) value is regarded as evidence of electrophilic reactivity likely to give rise to mutagenicity and carcinogenicity, then 13//14 (93%%) of the carcinogens are correctly identified by combination with the COMPACT procedure alone. It is possible, therefore, to establish likely carcinogenicity arising from either P450 mediation (CYP1 and CYP2E) or compound electrophilicity via the employment of a straightforward approach to molecular and electronic structure calculation, a process that can be performed in a relatively short time frame (i.e., less than 1 hour per chemical) and at a low cost.

Molecular modelling of the peroxisome proliferator-activated receptor α (PPARα) from human, rat and mouse, based on homology with the human PPARγ crystal structure

Abstract The generation of homology models of human, rat and mouse peroxisome proliferator-activated receptor α (PPARα) are reported, based on the recently published crystal structure of the human PPARγ ligand-binding domain (LBD) with bound ligand, rosiglitazone. It is found that a template of peroxisome proliferating fibrate drugs and related compounds can fit within the putative ligand-binding site of rat PPARα, via contacts with amino acid residues which are consistent with their biological potency for peroxisome proliferation, site-directed mutagenesis experiments and with quantitative structure–activity relationship (QSAR) analysis studies. The experimental binding affinity of leukotriene B 4 (LTB 4 ) for the mouse PPARα agrees closely with the calculated value based on the modelled interactions, whereas selective PPARα ligands such as clofibric acid are able to fit the human PPARα binding site in agreement with reported site-directed mutagenesis information.