QSAR Methods.

This chapter introduces the basis of computational chemistry and discusses how computational methods have been extended from physical to biological properties, and toxicology in particular, modeling. Since about three decades, chemical experimentation is more and more replaced by modeling and virtual experimentation, using a large core of mathematics, chemistry, physics, and algorithms. Animal and wet experiments, aimed at providing a standardized result about a biological property, can be mimicked by modeling methods, globally called in silico methods, all characterized by deducing properties starting from the chemical structures. Two main streams of such models are available: models that consider the whole molecular structure to predict a value, namely QSAR (quantitative structure-activity relationships), and models that check relevant substructures to predict a class, namely SAR. The term in silico discovery is applied to chemical design, to computational toxicology, and to drug discovery. Virtual experiments confirm hypotheses, provide data for regulation, and help in designing new chemicals.

Structures of Endocrine-Disrupting Chemicals Correlate with the Activation of 12 Classic Nuclear Receptors.

Endocrine-disrupting chemicals (EDCs) can inadvertently interact with 12 classic nuclear receptors (NRs) that disrupt the endocrine system and cause adverse effects. There is no widely accepted understanding about what structural features make thousands of EDCs able to activate different NRs as well as how these structural features exert their functions and induce different outcomes at the cellular level. This paper applies the hierarchical characteristic fragment methodology and high-throughput screening molecular docking to comprehensively explore the structural and functional features of EDCs for the 12 NRs based on more than 7000 chemicals from curated datasets. EDCs share three levels of key fragments. The primary and secondary fragments are associated with the binding of EDCs to four groups of receptors: steroidal nuclear receptors (SNRs, including androgen, estrogen, glucocorticoid, mineralocorticoid, and progesterone), retinoic acid receptors, thyroid hormone receptors, and vitamin D receptors. The tertiary fragments determine the activity type by interacting with two key locations in the ligand-binding domains of NRs (N-H5-H3-C and N-H7-H11-C for SNRs and N-H5-H5'-H2'-H3-C and N-H6'-H11-C for non-SNRs). The resulting compiled structural fragments of EDCs together with elucidated compound NR binding modes provide a framework for understanding the interactions between EDCs and NRs, facilitating faster and more accurate screening of EDCs for multiple NRs in the future.

QSAR modeling without descriptors using graph convolutional neural networks: the case of mutagenicity prediction.

Deep neural networks are effective in learning directly from low-level encoded data without the need of feature extraction. This paper shows how QSAR models can be constructed from 2D molecular graphs without computing chemical descriptors. Two graph convolutional neural network-based models are presented with and without a Bayesian estimation of the prediction uncertainty. The property under investigation is mutagenicity: Models developed here predict the output of the Ames test. These models take the SMILES representation of the molecules as input to produce molecular graphs in terms of adjacency matrices and subsequently use attention mechanisms to weight the role of their subgraphs in producing the output. The results positively compare with current state-of-the-art models. Furthermore, our proposed model interpretation can be enhanced by the automatic extraction of the substructures most important in driving the prediction, as well as by uncertainty estimations.

Structures of Endocrine-Disrupting Chemicals Determine Binding to and Activation of the Estrogen Receptor α and Androgen Receptor.

Endocrine-disrupting chemicals (EDCs) can interact with nuclear receptors, including estrogen receptor α (ERα) and androgen receptor (AR), to affect the normal endocrine system function, causing severe symptoms. Limited studies queried the EDC mechanisms, focusing on limited chemicals or a set of structurally similar compounds. It remained uncertain how hundreds of diverse EDCs could bind to ERα and AR and cause distinct functional consequences. Here, we employed a series of computational methodologies to investigate the structural features of EDCs that bind to and activate ERα and AR based on more than 4000 compounds. We used molecular docking and molecular dynamics simulations to elucidate the functional consequences and validated structure-function correlations experimentally using a time-resolved fluorescence resonance energy-transfer assay. We found that EDCs share three levels of key fragments. Primary (20 for ERα and 18 for AR) and secondary fragments (38 for ERα and 29 for AR) are responsible for the binding to receptors, and tertiary fragments determine the activity type (agonist, antagonist, or mixed). In summary, our study provides a general mechanism for the EDC function. Discovering the three levels of key fragments may drive fast screening and evaluation of potential EDCs from large sets of commercially used synthetic compounds.

Integrating in silico models and read-across methods for predicting toxicity of chemicals: A step-wise strategy.

In silico methods and models are increasingly used for predicting properties of chemicals for hazard identification and hazard characterisation in the absence of experimental toxicity data. Many in silico models are available and can be used individually or in an integrated fashion. Whilst such models offer major benefits to toxicologists, risk assessors and the global scientific community, the lack of a consistent framework for the integration of in silico results can lead to uncertainty and even contradictions across models and users, even for the same chemicals. In this context, a range of methods for integrating in silico results have been proposed on a statistical or case-specific basis. Read-across constitutes another strategy for deriving reference points or points of departure for hazard characterisation of untested chemicals, from the available experimental data for structurally-similar compounds, mostly using expert judgment. Recently a number of software systems have been developed to support experts in this task providing a formalised and structured procedure. Such a procedure could also facilitate further integration of the results generated from in silico models and read-across. This article discusses a framework on weight of evidence published by EFSA to identify the stepwise approach for systematic integration of results or values obtained from these "non-testing methods". Key criteria and best practices for selecting and evaluating individual in silico models are also described, together with the means to combining the results, taking into account any limitations, and identifying strategies that are likely to provide consistent results.

QSAR: What Else?

QSAR (quantitative structure-activity relationship) is a method for predicting the physical and biological properties of small molecules; it is today in large use in companies and public services. However, as any scientific method, it is nowadays challenged by more and more requests, especially considering its possible role in assessing the safety of new chemicals. Posing the question whether QSAR is a way not only to exploit available knowledge but also to build new knowledge, we shortly review QSAR history, thus searching for a QSAR epistemology. We consider the three pillars on which QSAR stands: biological data, chemical knowledge, and modeling algorithms. Most of the time we assume that biological data is a true picture of the world (as they result from good experimental practice), that chemical knowledge is scientifically true; so if a QSAR is not working, blame modeling. This opens the way to look at the role of modeling in developing scientific theories, and in producing knowledge. QSAR is a mature technology; however, debate is still active in many topics, in particular about the acceptability of the models and how they are explained. After an excursus in inductive reasoning, we relate the QSAR methodology to open debates in the philosophy of science.

QSAR Methods.

In this chapter, we introduce the basis of computational chemistry and discuss how computational methods have been extended to some biological properties and toxicology, in particular. Since about 20 years, chemical experimentation is more and more replaced by modeling and virtual experimentation, using a large core of mathematics, chemistry, physics, and algorithms. Then we see how animal experiments, aimed at providing a standardized result about a biological property, can be mimicked by new in silico methods. Our emphasis here is on toxicology and on predicting properties through chemical structures. Two main streams of such models are available: models that consider the whole molecular structure to predict a value, namely QSAR (Quantitative Structure Activity Relationships), and models that find relevant substructures to predict a class, namely SAR. The term in silico discovery is applied to chemical design, to computational toxicology, and to drug discovery. We discuss how the experimental practice in biological science is moving more and more toward modeling and simulation. Such virtual experiments confirm hypotheses, provide data for regulation, and help in designing new chemicals.

New clues on carcinogenicity-related substructures derived from mining two large datasets of chemical compounds.

In this study, new molecular fragments associated with genotoxic and nongenotoxic carcinogens are introduced to estimate the carcinogenic potential of compounds. Two rule-based carcinogenesis models were developed with the aid of SARpy: model R (from rodents' experimental data) and model E (from human carcinogenicity data). Structural alert extraction method of SARpy uses a completely automated and unbiased manner with statistical significance. The carcinogenicity models developed in this study are collections of carcinogenic potential fragments that were extracted from two carcinogenicity databases: the ANTARES carcinogenicity dataset with information from bioassay on rats and the combination of ISSCAN and CGX datasets, which take into accounts human-based assessment. The performance of these two models was evaluated in terms of cross-validation and external validation using a 258 compound case study dataset. Combining R and H predictions and scoring a positive or negative result when both models are concordant on a prediction, increased accuracy to 72% and specificity to 79% on the external test set. The carcinogenic fragments present in the two models were compared and analyzed from the point of view of chemical class. The results of this study show that the developed rule sets will be a useful tool to identify some new structural alerts of carcinogenicity and provide effective information on the molecular structures of carcinogenic chemicals.

Processing of surface EMG through pattern recognition techniques aimed at classifying shoulder joint movements.

Artificial arms for shoulder disarticulation need a high number of degrees of freedom to be controlled. In order to control a prosthetic shoulder joint, an intention detection system based on surface electromyography (sEMG) pattern recognition methods was proposed and experimentally investigated. Signals from eight trunk muscles that are generally preserved after shoulder disarticulation were recorded from a group of eight normal subjects in nine shoulder positions. After data segmentation, four different features were extracted (sample entropy, cepstral coefficients of the 4th order, root mean square and waveform length) and classified by means of linear discriminant analysis. The classification accuracy was 92.1% and this performance reached 97.9% after reducing the positions considered to five classes. To reduce the computational cost, the two channels with the least discriminating information were neglected yielding to a classification accuracy diminished by just 4.08%.

A new in silico classification model for ready biodegradability, based on molecular fragments.

Regulations such as the European REACH (Registration, Evaluation, Authorization and restriction of Chemicals) often require chemicals to be evaluated for ready biodegradability, to assess the potential risk for environmental and human health. Because not all chemicals can be tested, there is an increasing demand for tools for quick and inexpensive biodegradability screening, such as computer-based (in silico) theoretical models. We developed an in silico model starting from a dataset of 728 chemicals with ready biodegradability data (MITI-test Ministry of International Trade and Industry). We used the novel software SARpy to automatically extract, through a structural fragmentation process, a set of substructures statistically related to ready biodegradability. Then, we analysed these substructures in order to build some general rules. The model consists of a rule-set made up of the combination of the statistically relevant fragments and of the expert-based rules. The model gives good statistical performance with 92%, 82% and 76% accuracy on the training, test and external set respectively. These results are comparable with other in silico models like BIOWIN developed by the United States Environmental Protection Agency (EPA); moreover this new model includes an easily understandable explanation.

OCWLGI descriptors: theory and praxis.

The aim of this review is description of the logic and evolution of optimal descriptors OCWLGI calculated with the molecular graph and the demonstration of their ability as tools for the modeling of biological and physicochemical parameters of chemical compounds. The ability of optimal descriptors calculated with hydrogen suppressed graph (HSG), hydrogen filled graph (HFG) and graph of atomic orbitals (GAO) is demonstrated as a collection of quantitative structure-property relationships (QSPR) and quantitative structure-activity relationships (QSAR) for properties and endpoints available from the literature. The Monte Carlo method optimization of the correlation weights of local and global invariants (OCWLGI) of molecular graphs is used as the principle for building up descriptors which are discussed in this article. The statistical quality of the QSPR and QSAR models for physicochemical and biological properties which were obtained with the optimal descriptors are reasonably high.

QSAR as a random event: modeling of nanoparticles uptake in PaCa2 cancer cells.

Quantitative structure-property/activity relationships (QSPRs/QSARs) are a tool to predict various endpoints for various substances. The "classic" QSPR/QSAR analysis is based on the representation of the molecular structure by the molecular graph. However, simplified molecular input-line entry system (SMILES) gradually becomes most popular representation of the molecular structure in the databases available on the Internet. Under such circumstances, the development of molecular descriptors calculated directly from SMILES becomes attractive alternative to "classic" descriptors. The CORAL software (http://www.insilico.eu/coral) is provider of SMILES-based optimal molecular descriptors which are aimed to correlate with various endpoints. We analyzed data set on nanoparticles uptake in PaCa2 pancreatic cancer cells. The data set includes 109 nanoparticles with the same core but different surface modifiers (small organic molecules). The concept of a QSAR as a random event is suggested in opposition to "classic" QSARs which are based on the only one distribution of available data into the training and the validation sets. In other words, five random splits into the "visible" training set and the "invisible" validation set were examined. The SMILES-based optimal descriptors (obtained by the Monte Carlo technique) for these splits are calculated with the CORAL software. The statistical quality of all these models is good.

QSAR models for inhibitors of physiological impact of Escherichia coli that leads to diarrhea.

Quantitative structure - activity relationships (QSARs) developed to evaluate percentage of inhibition of STa-stimulated (Escherichia coli) cGMP accumulation in T84 cells are calculated by the Monte Carlo method. This endpoint represents a measure of biological activity of a substance against diarrhea. Statistical quality of the developed models is quite good. The approach is tested using three random splits of data into the training and test sets. The statistical characteristics for three splits are the following: (1) n=20, r(2)=0.7208, q(2)=0.6583, s=16.9, F=46 (training set); n=11, r(2)=0.8986, s=14.6 (test set); (2) n=19, r(2)=0.6689, q(2)=0.5683, s=17.6, F=34 (training set); n=12, r(2)=0.8998, s=12.1 (test set); and (3) n=20, r(2)=0.7141, q(2)=0.6525, s=14.7, F=45 (training set); n=11, r(2)=0.8858, s=19.5 (test set). Based on the proposed here models hypothetical compounds which can be useful agents against diarrhea are suggested.

CORAL: QSPR model of water solubility based on local and global SMILES attributes.

Water solubility is an important characteristic of a chemical in many aspects. However experimental definition of the endpoint for all substances is impossible. In this study quantitative structure-property relationships (QSPRs) for negative logarithm of water solubility-logS (mol L(-1)) are built up for five random splits into the sub-training set (≈55%), the calibration set (≈25%), and the test set (≈20%). Simplified molecular input-line entry system (SMILES) is used as the representation of the molecular structure. Optimal SMILES-based descriptors are calculated by means of the Monte Carlo method using the CORAL software (http://www.insilico.eu/coral). These one-variable models for water solubility are characterized by the following average values of the statistical characteristics: n(sub_train)=725-763; n(calib)=312-343; n(test)=231-261; r(sub_train)(2)=0.9211±0.0028; r(calib)(2)=0.9555±0.0045; r(test)(2)=0.9365±0.0073; s(sub_train)=0.561±0.0086; s(calib)=0.453±0.0209; s(test)=0.520±0.0205. Thus, the reproducibility of statistical quality of suggested models for water solubility confirmed for five various splits.

CORAL: binary classifications (active/inactive) for Liver-Related Adverse Effects of Drugs.

Classification data related to the Liver-Related Adverse Effects of Drugs have been studied with the CORAL software (http://www.insilico.eu/coral). Two datasets which contain compounds with two serum enzyme markers of liver toxicity: alanine aminotransferase (ALT, n=187) and aspartate aminotransferase (AST, n=209) are analyzed. Statistical quality of the prediction for ALT activity is n=35, Sensitivity = 0.5556, Specificity = 0.8077, and Accuracy = 0.7429. In the case of AST activity the prediction is characterized by n=42, Sensitivity = 0.6875, Specificity = 0.7692, and Accuracy = 0.7381. A number of structural alerts which can be related to the studied activities are revealed. It is the first attempt to build up the classification QSAR model by means of the Monte Carlo technique based on representation of the molecular structure by SMILES using the CORAL software.

Novel application of the CORAL software to model cytotoxicity of metal oxide nanoparticles to bacteria Escherichia coli.

Convenient to apply and available on the Internet software CORAL (http://www.insilico.eu/CORAL) has been used to build up quantitative structure-activity relationships (QSAR) for prediction of cytotoxicity of metal oxide nanoparticles to bacteria Escherichia coli (minus logarithm of concentration for 50% effect pEC50). In this study six random splits of the data into the training and test set were examined. It has been shown that the CORAL provides a reliable tool that could be used to build up a QSAR of the pEC50.

Calculation of molecular features with apparent impact on both activity of mutagens and activity of anticancer agents.

The analysis of the influence of molecular features which can be extracted from the simplified molecular input line entry system (SMILES) and involved in the process of the building up of a series of QSAR models (with different splits into training and test sets) by means of the CORAL software for mutagenicity and anticancer activity has been performed. The presence of nitrogen (sp3) is favorable for decrease of the both endpoints; the presence of only one ring is also promotor for decrease of the both endpoints; however the presence of two or three rings is favorable for increase of mutagenicity and decrease of anticancer activity. These findings provide useful criteria for further experimental and computational studies in the search for new anticancer agents.

QSAR models for toxicity of organic substances to Daphnia magna built up by using the CORAL freeware.

CORAL (CORrelations And Logic, http://www.insilico.eu/coral/) is a freeware available on the Internet. This freeware is designed to build up quantitative structure - property/activity relationships. The molecular structure for CORAL should be represented by the simplified molecular input line entry system (SMILES). Optimal descriptors calculated with SMILES are a mathematical function of the presence or absence of SMILES elements. The essence of this approach is the calculation of correlation weights for each element or combination of the elements by the Monte Carlo method. These coefficients serve to calculate the descriptors correlated with the endpoint for the training set, hoping that this correlation will also hold for the external test set. These descriptors can be improved by taking into account global physicochemical situations in molecules. An example of the physicochemical situation is the presence of oxygen and nitrogen. One can calculate these situations with SMILES and represent them by combining 0 (absence) and 1 (presence). The involving in the modelling of correlation weights of aforementioned physicochemical situations gave improvement in accuracy of models of toxicity to Daphnia magna for test set: n(test) = 75, r(2) = 0.7322, r(2) (pred) = 0.7193, r(2) (m) = 0.6549 (without correlation weights of the physicochemical situations); and n(test) = 75, r(2) = 0.7897, r(2) (pred) = 0.7790, r(2) (m) = 0.6850 (with aforementioned correlation weights of physicochemical situations).

SMILES-based QSAR approaches for carcinogenicity and anticancer activity: comparison of correlation weights for identical SMILES attributes.

CORAL software (http://www.insilico.eu/coral/) has been used for modeling of carcinogenicity (logTD50) of 401 compounds, and anticancer activity (-logIC50) of 100 compounds, on the basis of quantitative structure activity relationships (QSAR). The simplified molecular input line entry system (SMILES) was used for the representation of the molecular structures. A new additional global invariant of the molecular structure was tested. This is an indicator of the presence of pairs of chemical elements (F, Cl, Br, N, O, S, and P). Three random splits into sub-training, calibration, and test set were examined. Molecular features (calculated with SMILES and statistically significant), which increase the anticancer activity have been identified: their presence in the molecular structure could be helpful criterion in the search for new anticancer agents.