Ying Ping Zhang

Development, Characterization and Application of Predictive-toxicology Models

The adoption of SAR techniques for risk assessment purposes requires that the predictive performance of models be characterized and optimized. The development of such methods with respect to CASE/MULTICASE are described. Moreover, the effects of size, informational content, ratio of actives/inactives in the model on predictivity must be determined. Characterized models can provide mechanistic insights: nature of toxicophore, reactivity, receptor binding. Comparison of toxicophores among SAR models allows a determination of mechanistic overlaps (e.g., mutagenicity, toxicity, inhibition of gap junctional intercellular communication vs. carcinogenicity). Methods have been developed to combine SAR submodels and thereby improve predictive performance. Now that predictive toxicology methods are gaining acceptance, the development of Good Laboratory Practices is a further priority, as is the development of graduate programs in Computational Toxicology to adequately train the needed professional.

Identification of 'genotoxic' and 'non-genotoxic' alerts for cancer in mice: the carcinogenic potency database.

A set of chemicals tested for carcinogenicity in mice that have been analyzed by Gold et al. [L.S. Gold, C.B. Sawyer, R. Magaw, G.M. Backman, M. deVeciana, R. Levinson, N.K. Hooper, W.R. Havender, L. Bernstein, R. Peto, M.C. Pike, B.N. Ames, Environ. Health Perspect. 58 (1984) 9-319; L.S. Gold, M. deVeciana, G.M. Backman, M. Lopipero, M. Smith, R. Blumenthal, R. Levinson, L. Bernstein, B.N. Ames, Environ. Health Perspect. 67 (1986) 161-200; L.S. Gold, T.H. Slone, G.M. Backman, R. Magaw, M. DaCosta, P. Lopipero, M. Blumenthal, B.N. Ames, Environ. Health Perspect. 74 (1987) 237-329; L.S. Gold, T.H. Slone, G.M. Backman, S. Eisenberg, M. DaCosta, M. Wong, N.B. Manley, L. Rohrbach, B.N. Ames, Environ. Health Perspect. 84 (1990) 215-286; L.S. Gold, N.B. Manley, T.H. Slone, T.H. Garfinkle, L. Rohrbach, B.N. Ames, Environ. Health Perspect. 100 (1993) 65-135] in the first five plots of the carcinogenic potency database (CPDB) was subjected to CASE/MULTICASE analyses. Briefly, CASE/MULTICASE is a computer-automated structure evaluation system that is capable of identifying structural features of chemicals associated with a specified biological activity (e.g., carcinogenicity or mutagenicity). These features are then incorporated into a structure-activity relationship (SAR) model for the analyzed database. The mouse CPDB used in this study consists of 627 chemicals, 289 of which are carcinogens, 11 marginal or weak carcinogens (i.e., chemicals requiring high doses to induce cancer) and 327 non-carcinogens. In an internal prediction analysis where the CASE/MULTICASE SAR model was used to predict the carcinogenicity of chemicals used to create the model, a concordance between experimental and predicted results of 96% was obtained. This indicates that the model is able to satisfactorily explain the chemicals in the learning set. In a drop-one cross-validation study where chemicals were removed one at a time and the remaining n - 1 chemicals were used in an iterative method to create a model to predict the removed chemical, CASE/MULTICASE was able to achieve a concordance between experimental and predicted results of 70%. Using a modified validation process designed to investigate the predictivity of a more focused SAR model, the system achieved a 78% concordance between experimental and predicted results. Among the major biophores identified by CASE/MULTICASE associated with cancer causation in mice several are derived from electrophilic or potentially electrophilic compounds (e.g., hydrazines, N-mustards, N-nitrosamines, aromatic amines, reactive halogens, and quinones). Other biophores however are derived from chemicals seemingly devoid of actual or potential DNA-reactivity and as such may represent structural feature of non-genotoxic carcinogens.

Modeling the mouse lymphoma forward mutational assay: the Gene-Tox program database.

An SAR model of the induction of mutations at the tk(+/-) locus of L5178Y mouse lymphoma cells (MLA, for mouse lymphoma assay) was derived based upon a re-evaluation of experimental results reported by a Gene-Tox (GT) working group [A.D. Mitchell, A.E. Auletta, D. Clive, P.E. Kirby, M.M. Moore, B.C. Myhr, The L5178Y/tk(+/-) mouse lymphoma specific gene and chromosomal mutation assay. A phase III report of the U.S. Environmental Protection Agency Gene-Tox Program, Mutation Res. 394 (1997) 177-303.]. The predictive performance of the GT MLA SAR model was similar to that of a Salmonella mutagenicity model containing the same number of chemicals. However, the structural determinants (biophores) derived from the GT MLA SAR model include both electrophilic as well as non-electrophilic moieties, suggesting that the induction of mutations in the MLA may occur by both direct interaction with DNA and by non-DNA-related mechanisms. This was confirmed by the observation that the set of biophores associated with MLA overlapped significantly with those associated with phenomena related to loss of heterozygosity, chromosomal rearrangements and aneuploidy. The MLA SAR model derived from the GT data evaluation was significantly more predictive than an SAR model previously derived from MLA data reported by the US National Toxicology Program [B. Henry, S.G. Grant, G. Klopman, H.S. Rosenkranz, Induction of forward mutations at the thymidine kinase locus of mouse lymphoma cells: evidence for electrophilic and non-electrophilic mechanisms, Mutation Res. 397 (1998) 331-335.]. Moreover, the latter model appeared to be more complex than the former, suggesting that the GT induction data was both simpler mechanistically and more homogeneous than that of the NTP.

Structural determinants of developmental toxicity in hamsters.

A CASE/MULTICASE structure activity relationship (SAR) model of developmental toxicity of chemicals in hamsters (HaDT) was developed. The model exhibited a predictive performance of 74%. The models overall predictivity and informational content were similar to those of an SAR model of mutagenicity in Salmonella. However, unlike the Salmonella mutagenicity model, the HaDT model did not identify overtly chemically reactive moieties as associated with activity. Moreover, examination of the number and nature of significant structural determinants suggested that developmental toxicity in hamsters was not the result of a unique mechanism or attack on a specific molecular target. The analysis also indicated that the availability of experimental data on additional chemicals would improve the performance of the SAR model.

Applications of the CASE/MULTICASE SAR Method to Environmental and Public Health Situations

The availability of validated and characterized SAR models of toxicological phenomena provides a method to apply SAR technology to a variety of environmental, public health and industrial situations. These include (i) the prioritization of environmental pollutants for control and/or regulation, (ii) the design of multi-action optimized therapeutics from which the potential for unwanted side-effects have been engineered out, (iii) the development of SAR-based computer-driven screening procedure to identify candidate therapeutics based upon combinatorial chemistry or compilations of molecular structures, (iv) the generation of toxicological profiles to be used in the selection of benign chemicals in the early stages of product development.

Implications of newly recognized relationships between mutagenicity, genotoxicity and carcinogenicity of molecules

The CASE structure-activity relational method was used to predict the mutagenicity, cytogenotoxicity, carcinogenicity, sensory irritation, male rat-specific alpha 2 mu-nephrotoxicity and maximum tolerated dose of a population of molecules (N greater than or equal to 1300). These chemicals were then sorted out by their predicted responses to specific tests and sub-populations of molecules with different prevalence with respect to described endpoints were constructed, i.e. 0-100% prevalences of mutagens, rodent carcinogens and SCE inducers. The predicted properties of these populations were analyzed and the overlap among tests was determined. The method also permits the determination of the dependence among assays and the level of false-positive and false-negative predictions.

Human developmental toxicity and mutagenesis

A previously described SAR model of human developmental toxicity was analyzed further. The model shows a number of mechanistic similarities with SAR models of other toxicological phenomena (systemic toxicity, chromosomal and genomic effects). This implies that there are many targets associated with developmental effects. Surprisingly the analyses revealed no significant mechanistic overlap between developmental toxicity in humans and mutagenicity in Salmonella, a surrogate for the occurrence of point mutations. Our study indicates that this lack of similarity is likely the result of the pre-screening strategies which largely eliminate Salmonella mutagens from among the therapeutics introduced into human medicine.

PREDICTION OF THE METABOLISM AND TOXICOLOGICAL PROFILES OF GASOLINE OXYGENATES

Methyl tert -butyl ether (MTBE), ethyl tert -butyl ether (ETBE), tert -amyl methyl ether (TAME), and diisopropyl ether (DIPE) were evaluated with CASE/MULTICASE structure relational models in order to determine their potential to pose human health risks. None of the parent ethers were predicted to be sensory irritants, eye irritants, contact sensitizers, mutagens, developmental toxicants, or carcinogens. The putative metabolites of ETBE were generated by META, an expert system, and evaluated for their potential to contribute to toxicity. Several of the metabolites were predicted by CASE/MULTICASE to be sensory irritants, contact sensitizers, mutagens, developmental toxicants, and carcinogens. A preliminary examination of the putative metabolites of TAME and DIPE revealed the presence of epoxides, a class of chemicals associated with developmental toxicity, carcinogenicity, and dermal contact sensitivity.

Structural Features Contributing to Biodegradability

In this review it is shown that biodegradation of organic molecules by mixed bacterial cultures can be modelled successfully by an expert structure-activity relational system.