Crossing the Halfway Point: Aptamer-Based, Highly Multiplexed Assay for the Assessment of the Proteome.

Measuring responses in the proteome to various perturbations improves our understanding of biological systems. The value of information gained from such studies is directly proportional to the number of proteins measured. To overcome technical challenges associated with highly multiplexed measurements, we developed an affinity reagent-based method that uses aptamers with protein-like side chains along with an assay that takes advantage of their unique properties. As hybrid affinity reagents, modified aptamers are fully comparable to antibodies in terms of binding characteristics toward proteins, including epitope size, shape complementarity, affinity and specificity. Our assay combines these intrinsic binding properties with serial kinetic proofreading steps to allow highly effective partitioning of stable specific complexes from unstable nonspecific complexes. The use of these orthogonal methods to enhance specificity effectively overcomes the severe limitation to multiplexing inherent to the use of sandwich-based methods. Our assay currently measures half of the unique proteins encoded in the human genome with femtomolar sensitivity, broad dynamic range and exceptionally high reproducibility. Using machine learning to identify patterns of change, we have developed tests based on measurement of multiple proteins predictive of current health states and future disease risk to guide a holistic approach to precision medicine.

Inhibition of the Complement Alternative Pathway by Chemically Modified DNA Aptamers That Bind with Picomolar Affinity to Factor B.

The complement system is a conserved component of innate immunity that fulfills diverse roles in defense and homeostasis. Inappropriate activation of complement contributes to many inflammatory diseases, however, which has led to a renewed emphasis on development of therapeutic complement inhibitors. Activation of complement component C3 is required for amplification of complement and is achieved through two multisubunit proteases called C3 convertases. Of these, the alternative pathway (AP) C3 convertase is responsible for a majority of the C3 activation products in vivo, which renders it an attractive target for inhibitor discovery. In this study, we report the identification and characterization of two related slow off-rate modified DNA aptamers (SOMAmer) reagents that inhibit formation of the AP C3 convertase by binding to the proprotease, factor B (FB). These aptamers, known as SL1102 (31 bases) and SL1103 (29 bases), contain uniform substitutions of 5-(N-2-naphthylethylcarboxyamide)-2'-deoxyuridine for deoxythymidine. SL1102 and SL1103 bind FB with K d values of 49 and 88 pM, respectively, and inhibit activation of C3 and lysis of rabbit erythrocytes under AP-specific conditions. Cocrystal structures of SL1102 (3.4 Å) and SL1103 (3.1 Å) bound to human FB revealed that SL1102 and SL1103 recognize a site at the juncture of the CCP1, CCP3, and vWF domains of FB. Consistent with these structures and previously published information, these aptamers inhibited FB binding to C3b and blocked formation of the AP C3 convertase. Together, these results demonstrate potent AP inhibition by modified DNA aptamers and expand the pipeline of FB-binding molecules with favorable pharmacologic properties.

Estimating evaporation rates and contaminant air concentrations due to small spills of non-ideal aqueous organic solvent mixtures in a controlled environment.

Although small spills of non-ideal organic solvent mixtures are ubiquitous undesirable events in occupational settings, the potential risk of exposure associated with such scenarios remains insufficiently investigated. This study aimed to examine the impact of non-ideality on evaporation rates and contaminant air concentrations resulting from small spills of organic solvent mixtures. Evaporation rate constants alphas (α) were experimentally measured for five pure solvents using a gravimetric approach during solvent evaporation tests designed to simulate small spills of solvents. Two equations were used for estimating contaminants' evaporation rates from aqueous mixtures assuming either ideal or non-ideal behavior based on the pure-chemical alpha values. A spill model also known as the well-mixed room model with exponentially decreasing emission rate was used to predict air concentrations during various spill scenarios based on the two sets of estimated evaporation rates. Model predictive performance was evaluated by comparing the estimates against real-time concentrations measured for the same scenarios. Evaluations for 12 binary non-ideal aqueous mixtures found that the estimated evaporation rates accounting for the correction by the activity coefficients of the solvents (median = 0.0318 min-1) were higher than the evaporation rates estimated without the correction factor (median = 0.00632 min-1). Model estimates using the corrected evaporation rates reasonably agreed with the measured values, with a median predicted peak concentrations-to-measured peak concentrations ratio of 0.92 (0.81 to 1.32) and a median difference between the predicted and the measured peak times of -5 min. By contrast, when the non-corrected evaporation rates were used, the median predicted peak concentrations-to-measured peak concentrations ratio was 0.31 (0.08 to 0.75) and the median difference between the predicted and the measured peak times was +33 min. Results from this study demonstrate the importance of considering the non-ideality effect for accurately estimating evaporation rates and contaminant air concentrations generated by solvent mixtures. Moreover, this study is a step further in improving knowledge of modeling exposures related to small spills of organic solvent mixtures.

Predicting first-order evaporation rate constant alpha (α) from small spills of organic solvents in a controlled environment.

Exposures to vapors generated by small spills of organic solvents are common in the occupational hygiene practice. In these scenarios, contaminant mass release is exponentially decreasing, driven by an evaporation rate constant alpha (α). Knowing α is fundamental for adequately modeling peak concentrations and/or short-term exposures that occur and for achieving efficient occupational risk analysis and management. The purpose of this study was to measure alpha experimentally using a gravimetric approach in a controlled environment during solvent evaporation tests designed to simulate small spills of solvents. The effects of several factors on α were evaluated. Equations based on regression models derived from the experimental data were proposed for predicting α. Predictions were externally validated against experimental data. A total of 183 tests was performed. Data analyses found that alpha (α) values increased with vapor pressure, spill surface area-to-spill volume ratio, and air speed across the spill. Larger α were associated with petri dish containers compared to watch glasses. Three regression models were created for predicting α. They had four variables in common, namely vapor pressure, molecular weight, air speed above the liquid, and surface tension of the liquid. The fifth variable was either spill volume, spill surface area, or spill surface area-to-spill volume ratio. The R2 of the regression models were equal to 0.98. External validation showed mean relative errors of -32.9, -32.0, and -25.5%, respectively, with associated standard deviations of the relative errors of 17.7, 33.3, and 26.0%, respectively, and associated R2 of 0.92, 0.65, and 0.87, respectively. The proposed equations can be used for estimating α in exposure scenarios similar to those evaluated in this study. Moreover, these models constitute a step further in the improvement of knowledge on estimating evaporation rates for small spills of organic solvents.

Modeling occupational exposure to solvent vapors using the Two-Zone (near-field/far-field) model: a literature review.

The Two-Zone model is used in occupational hygiene to predict both near-field and far-field airborne contaminant concentrations. A literature review was carried out on 21 scientific publications in which the Two-Zone model was used to assess occupational exposure to solvent vapors. Data on exposure scenarios, solvents, generation/emission rates, near- and far-field parameters, and model performance were collected and analyzed. Over the 24 exposure scenarios identified, 18 were evaluated under controlled conditions, 5 under normal workplace activities, and 1 was reported based on literature data. The scenarios involved a variety of tasks which consisted, mostly, of cleaning metal parts, spraying solvents onto surfaces, spilling liquids, and filling containers with volatile substances. Twenty-eight different solvents were modeled and the most commonly tested were benzene, toluene, and acetone. Emission rates were considered constant in 16 scenarios, exponentially decreasing in 6 scenarios, and intermittent in 2 scenarios. Four-hundred-and-forty-six (446) predicted-to-measured concentration ratios were calculated across the 21 studies; 441 were obtained in controlled conditions, 4 under normal workplace activities, and 1 was calculated based on the literature data. For controlled studies, the Two-Zone model predictive performance was within a factor of 0.3-3.7 times the measured concentrations with 93% of the values between 0.5 and 2. The model overestimated the measured concentrations in 63% of the evaluations. The median predicted concentration for the near-field was 1.38 vs. 1.02 for the far-field. Results suggest that the model might be a useful tool for predicting occupational exposure to vapors of solvents by providing a conservative approach. Harmonization in model testing strategies and data presentation is needed in future studies to improve the assessment of the predictability of the Two-Zone model. Moreover, this review has provided a database of exposure scenarios, input parameter values, and model predictive performances which can be useful to occupational hygienists in their future modeling activities.

Using checklists and algorithms to improve qualitative exposure judgment accuracy.

Most exposure assessments are conducted without the aid of robust personal exposure data and are based instead on qualitative inputs such as education and experience, training, documentation on the process chemicals, tasks and equipment, and other information. Qualitative assessments determine whether there is any follow-up, and influence the type that occurs, such as quantitative sampling, worker training, and implementing exposure and risk management measures. Accurate qualitative exposure judgments ensure appropriate follow-up that in turn ensures appropriate exposure management. Studies suggest that qualitative judgment accuracy is low. A qualitative exposure assessment Checklist tool was developed to guide the application of a set of heuristics to aid decision making. Practicing hygienists (n = 39) and novice industrial hygienists (n = 8) were recruited for a study evaluating the influence of the Checklist on exposure judgment accuracy. Participants generated 85 pre-training judgments and 195 Checklist-guided judgments. Pre-training judgment accuracy was low (33%) and not statistically significantly different from random chance. A tendency for IHs to underestimate the true exposure was observed. Exposure judgment accuracy improved significantly (p <0.001) to 63% when aided by the Checklist. Qualitative judgments guided by the Checklist tool were categorically accurate or over-estimated the true exposure by one category 70% of the time. The overall magnitude of exposure judgment precision also improved following training. Fleiss' κ, evaluating inter-rater agreement between novice assessors was fair to moderate (κ = 0.39). Cohen's weighted and unweighted κ were good to excellent for novice (0.77 and 0.80) and practicing IHs (0.73 and 0.89), respectively. Checklist judgment accuracy was similar to quantitative exposure judgment accuracy observed in studies of similar design using personal exposure measurements, suggesting that the tool could be useful in developing informed priors and further demonstrating its usefulness in producing accurate qualitative exposure judgments.

Creatinine and specific gravity normalization in biological monitoring of occupational exposures.

Reference values for the biological monitoring of occupational exposures are generally normalized on the basis of creatinine (CR) concentration or specific gravity (SG) to account for fluctuations in urine dilution. For instance, the American Conference of Governmental Industrial Hygienists (ACGIH(®)) uses a reference value of 1g/L for CR. The comparison of urinary concentrations of biomarkers between studies requires the adjustment of results based on a reference CR and/or SG value, although studies have suggested that age, sex, muscle mass, and time of the day can exert non-negligible influences on CR excretion, while SG appears to be less affected. The objective of this study was to propose reference values for urinary CR and SG based on the results of samples sent for analysis by occupational health practitioners to the laboratory of the Occupational Health and Safety Research Institute of Québec (IRSST). We analyzed a database containing 20,395 urinary sample results collected between 1985 and 2010. Linear mixed-effects models with worker as a random effect were used to estimate the influence of sex and collection period on urinary CR and SG. Median CR concentrations were 25-30% higher in men (1.6 g/L or 14.4 mmol/L) than in women (1.2 g/L or 10.2 mmol/L). Four percent of the samples for men and 12% for women were below the acceptable threshold for CR (4.4 mmol/L). For SG, 5% of samples for men and 12% for women were below the threshold of 1.010. The difference in SG levels between sexes was lower than for CR, with a median of 1.024 for men compared to 1.020 for women. Our results suggest that the normalization of reference values based on a standard CR value of 1 g/L as proposed by the ACGIH is a conservative approach. According to the literature, CR excretion is more influenced by physiological parameters than SG. We therefore suggest that correction based on SG should be favored in future studies involving the proposal of reference values for the biological monitoring of occupational exposures.

Dermal absorption of chemicals: estimation by IH SkinPerm.

This article describes the IH SkinPerm mathematical tool for estimating dermal absorption. The first part provides the scientific background of the IH SkinPerm model, including the QSARs and the developed differential equations. Then the practical value of the tool is demonstrated through example dermal absorption assessments for substances with skin notations. IH SkinPerm simulates three types of dermal absorption scenarios relevant to occupational environments. The first is dermal absorption from instantaneous splash type exposures onto bare skin for pure liquids. The second estimates dermal absorption from the deposition of pure liquids over time. The third enables estimation of dermal uptake from an airborne vapor concentration. A simulation with IH SkinPerm was made using vapor absorption data published from volunteer exposure studies. Comparison of measured and estimated dermal absorbed dose showed IH SkinPerm estimated dermal absorbed dose was within a factor of 3 compared to the reported study values. IH SkinPerm accounts for substance volatility and evaporated mass and provides real-time description of dermal absorption with graphical displays and numerical outputs. To assess absorption resulting from dermal exposure scenarios, the mass of the substance loaded onto the skin, substance physical chemical properties, exposure duration, and the skin surface area affected are the only required input parameters.

Comparing Antoine parameter sources for accurate vapor pressure prediction across a range of temperatures.

Determining the vapor pressure of a substance at the relevant process temperature is a key component in conducting an exposure assessment to ascertain worker exposure. However, vapor pressure data at various temperatures relevant to the work environment is not readily available for many chemicals. The Antoine equation is a mathematical expression that relates temperature and vapor pressure. The objective of this analysis was to compare Antoine parameter data from 3 independent data sources; Hansen, Yaws, and Custom data and identify the source that generates the most accurate vapor pressure values with the least bias, relative to the referent data set from the CRC Handbook of Chemistry and Physics. Temperatures predicted from 3 different Antoine sources across a range of vapor pressures for 59 chemicals are compared to the reference source. The results show that temperatures predicted using Antoine parameters from the 3 sources are not statistically significantly different, indicating that all 3 sources could be useful. However, the Yaws dataset will be used in the SDM 2.0 because the data is readily available and robust.

Plasma proteome of growing tumors.

Early detection of cancer is vital for the best chance of successful treatment, but half of all cancers are diagnosed at an advanced stage. A simple and reliable blood screening test applied routinely would therefore address a major unmet medical need. To gain insight into the value of protein biomarkers in early detection and stratification of cancer we determined the time course of changes in the plasma proteome of mice carrying transplanted human lung, breast, colon, or ovarian tumors. For protein measurements we used an aptamer-based assay which simultaneously measures ~ 5000 proteins. Along with tumor lineage-specific biomarkers, we also found 15 markers shared among all cancer types that included the energy metabolism enzymes glyceraldehyde-3-phosphate dehydrogenase, glucose-6-phophate isomerase and dihydrolipoyl dehydrogenase as well as several important biomarkers for maintaining protein, lipid, nucleotide, or carbohydrate balance such as tryptophanyl t-RNA synthetase and nucleoside diphosphate kinase. Using significantly altered proteins in the tumor bearing mice, we developed models to stratify tumor types and to estimate the minimum detectable tumor volume. Finally, we identified significantly enriched common and unique biological pathways among the eight tumor cell lines tested.

A nonclinical safety and pharmacokinetic evaluation of N6022: a first-in-class S-nitrosoglutathione reductase inhibitor for the treatment of asthma.

S-nitrosoglutathione reductase is the primary enzyme responsible for the metabolism of S-nitrosoglutathione (GSNO), the body's main source of bioavailable nitric oxide. Through its catabolic activity, GSNO reductase (GSNOR) plays a central role in regulating endogenous S-nitrosothiol levels and protein S-nitrosation-based signaling. By inhibiting GSNOR, we aim to increase pulmonary GSNO and induce bronchodilation while reducing inflammation in lung diseases such as asthma. To support the clinical development of N6022, a first-in-class GSNOR inhibitor, a 14-day toxicology study was conducted. Sprague-Dawley rats were given 2, 10 or 50 mg/kg/day N6022 via IV administration. N6022 was well tolerated at all doses and no biologically significant adverse findings were noted in the study up to 10 mg/kg/day. N6022-related study findings were limited to the high dose group. One male rat had mild hepatocellular necrosis with accompanying increases in ALT and AST and several male animals had histological lung assessments with a slight increase in foreign body granulomas. Systemic exposure was greater in males than females and saturation of plasma clearance was observed in both sexes in the high dose group. Liver was identified as the major organ of elimination. Mechanistic studies showed dose-dependent effects on the integrity of a rat hepatoma cell line.

Phase I, pharmacokinetic and biological correlative study of OSI-7904L, a novel liposomal thymidylate synthase inhibitor, and cisplatin in patients with solid tumors.

PURPOSE: To evaluate the safety and describe the pharmacokinetic profile of OSI-7904L, a novel liposomal thymidylate synthase inhibitor, in combination with cisplatin (CDDP) in adults with advanced solid tumors. EXPERIMENTAL DESIGN: CDDP was administered as a 2-h intravenous infusion followed by OSI-7904L intravenously over 30 min, both given every 3 weeks. Doses of each drug were escalated in separate cohorts of patients. Five dose levels of CDDP/OSI-7904L were explored: 60/6, 60/9, 60/12, 60/7.5, and 75/7.5 mg/m2. Pharmacokinetic samples, baseline plasma homocysteine, and genotype polymorphisms were evaluated. RESULTS: Twenty-seven patients were treated with 101 total courses of CDDP/OSI-7904L. Dose-limiting toxicity was observed in 2 patients in the CDDP/OSI-7904L 60/12 mg/m2 cohort. One patient experienced rash, stomatitis, dehydration, renal failure, hyperbilirubinemia, and fatal neutropenic sepsis, whereas the other patient experienced grade 3 nausea, vomiting, and ileus. Therefore, the CDDP/OSI-7904L 60/9 mg/m2 cohort was expanded, with 2 of 6 patients reporting significant fatigue. Other toxicities were mild or moderate. Intermediate dose levels of 60/7.5 and 75/7.5 mg/m2 were evaluated, and the latter was identified as the recommended dose for phase II studies. No major pharmacokinetic interactions between CDDP and OSI-7904L were observed. Three patients had partial responses (gastric adenocarcinoma and heavily pretreated breast cancer). There was no significant relationship between baseline homocysteine and toxicity. CONCLUSIONS: The recommended doses for CDDP and OSI-7904L administered once every 3 weeks are 75 and 7.5 mg/m2, respectively. Pharmacokinetic interaction between the agents was not apparent. Preliminary clinical activity was observed in breast and gastric cancer.

Proteomic Profiles in Advanced Age-Related Macular Degeneration Using an Aptamer-Based Proteomic Technology.

PURPOSE: To explore top-ranked plasma proteins related to neovascular age-related macular degeneration (AMD) and geographic atrophy (GA), and explore pathways related to neovascular AMD and GA. METHODS: We conducted a pilot study of patients with neovascular AMD (n = 10), GA (n = 10), and age-matched cataract controls (n = 10) who were recruited into an AMD registry. We measured 4001 proteins in ethylenediaminetetraacetic acid plasma samples using an aptamer-based proteomic technology. Relative concentrations of each of 4001 proteins were log (base 2) transformed and compared between cases of neovascular AMD and GA versus controls using linear regression. Pathway analysis was conducted using pathways downloaded from Reactome. RESULTS: In this pilot study, higher levels of vinculin and lower levels of CD177 were found in patients with neovascular AMD compared with controls. Neuregulin-4 was higher and soluble intercellular adhesion molecule-1 was lower in patients with GA compared with controls. For neovascular AMD, cargo trafficking to the periciliary membrane, fibroblast growth factor receptor 3b ligand binding and activation, and vascular endothelial growth factor-related pathways were in the top ranked pathways. The top-ranked pathways for GA included several related to ErbB4 signaling. CONCLUSIONS: We found different proteins and different pathways associated with neovascular AMD and GA. Vinculin and some of the top-ranked pathways have been previously associated with AMD, whereas others have not been described. TRANSLATIONAL RELEVANCE: Biomarkers identified in plasma likely reflect systemic alterations in protein expression and may improve our understanding of the mechanisms leading to AMD.

The effect of the number of counted traverses on the estimation of the total spore count sampled on a non-cultivable slit impactor.

Various counting rules are used for spore trap analysis. Partial count can lead to concentration errors. This paper demonstrates that the number of traverses counted affects the final results.

A phase I study of a new nucleoside analogue, OSI-7836, using two administration schedules in patients with advanced solid malignancies.

PURPOSE: To investigate the safety, tolerability, and pharmacokinetic profile of the novel nucleoside analogue OSI-7836 in patients with advanced solid malignancies. EXPERIMENTAL DESIGN: OSI-7836 was initially given as a 60-minute i.v. infusion on day 1 every 21 days. In view of its dose-limiting toxicities, the administration time was amended to a 5-minute bolus, and subsequently, the schedule was amended to weekly for 4 weeks followed by a 2-week rest. Blood and urine samples were collected for pharmacokinetic studies. Analyses of cytokines and lymphocyte subsets were added later in the study to elucidate a mechanism for the severe fatigue and lymphocyte depletion observed in earlier patients. RESULTS: Thirty patients received a total of 61 treatment cycles. Fatigue was the main dose-limiting toxicity. Maximum-tolerated dose was defined as 300 mg/m2 in the 60-minute infusion, (three times per week) schedule; 400 mg/m2 in the 5-minute bolus infusion, (three times per week) schedule; and 100 mg/m2 in the weekly schedule. Other common toxicities were nausea, vomiting, rash, fever, and a flu-like syndrome. There were no clinically significant hematologic toxicities. Following the initial dose, OSI-7836 was eliminated from plasma with a median (range) elimination half-life of 48.3 minutes (22.6-64.8 minutes). Lymphocyte subset analysis showed a significant drop in B cell counts, which persisted to day 14 and beyond. Cytokine analysis showed significant elevations of interleukin-6 and interleukin-10 in all patients who received > or = 200 mg/m2 OSI-7836. Best response was disease stabilization in seven patients. CONCLUSION: OSI-7836 was associated with excessive fatigue, and despite changes in its schedule and duration of administration, we did not observe an improvement in its tolerability. Its potentially selective effect on B lymphocytes could be exploited in further studies in specific hematologic malignancies.

Pharmacokinetic Properties of DNA Aptamers with Base Modifications.

The addition of novel side chains at the 5-position of uracil is an effective means to increase chemical diversity of aptamers and hence the success rate for discovery of high-affinity ligands to protein targets. Such modifications also increase nuclease resistance, which is useful in a range of applications, especially for therapeutics. In this study, we assess the impact of these side chains on plasma pharmacokinetics of modified aptamers conjugated to a 40 kDa polyethylene glycol. We show that clearance from plasma depends on relative hydrophobicity: side chains with a negative cLogP (more hydrophilic) result in slower plasma clearance compared with side chains with a positive cLogP (more hydrophobic). We show that clearance increases with the number of side chains in sequences of ≥28 synthons, but this effect is dramatically diminished in shorter sequences. These results serve as a guide for the design of new therapeutic aptamers with diversity-enhancing side chains.

Fit for the Eye: Aptamers in Ocular Disorders.

For any new class of therapeutics, there are certain types of indications that represent a natural fit. For nucleic acid ligands in general, and aptamers in particular, the eye has historically been an attractive site for therapeutic intervention. In this review, we recount the discovery and early development of three aptamers designated for use in ophthalmology, one approved (Macugen), and two in late-stage development (Fovista and Zimura). Every one of these molecules was originally intended for other indications. Key improvements in technology, specifically with regard to libraries used for in vitro selection and subsequent chemical optimization of aptamers, have played an important role in allowing the identification of development candidates with suitable properties. The lessons learned from the selection of these molecules are valuable for informing us about the many remaining opportunities for aptamer-based therapeutics in ophthalmology as well as for identifying additional indications for which aptamers as a class of therapeutics have distinct advantages.

Chemically Modified Interleukin-6 Aptamer Inhibits Development of Collagen-Induced Arthritis in Cynomolgus Monkeys.

Interleukin-6 (IL-6) is a potent mediator of inflammatory and immune responses, and a validated target for therapeutic intervention of inflammatory diseases. Previous studies have shown that SL1026, a slow off-rate modified aptamer (SOMAmer) antagonist of IL-6, neutralizes IL-6 signaling in vitro. In the present study, we show that SL1026 delays the onset and reduces the severity of rheumatoid symptoms in a collagen-induced arthritis model in cynomolgus monkeys. SL1026 (1 and 10 mg/kg), administered q.i.d., delayed the progression of arthritis and the concomitant increase in serum IL-6 levels compared to the untreated control group. Furthermore, SL1026 inhibited IL-6-induced STAT3 phosphorylation ex vivo in T lymphocytes from human blood and IL-6-induced C-reactive protein and serum amyloid A production in human primary hepatocytes. Importantly, SOMAmer treatment did not elicit an immune response, as evidenced by the absence of anti-SOMAmer antibodies in plasma of treated monkeys. These results demonstrate that SOMAmer antagonists of IL-6 may be attractive agents for the treatment of IL-6-mediated diseases, including rheumatoid arthritis.

Database for the toxicological evaluation of mixtures in occupational atmospheres.

Workers are regularly simultaneously exposed to multiple chemical substances. As in the ACGIH (American Conference of Governmental Industrial Hygienists) approach, the Québec Regulation prescribes that when two or more hazardous substances are present in workplaces and have similar effects on the same organs of the human body, their effects should be considered additive, unless established otherwise. This project was undertaken to develop a user-friendly toxicological database aid in identification of possible interactive effects of mixtures present in the work environment. In the first phase of the project, standard general literature references were used to compile critical data, such as target organs, effects on the target organs, mechanisms of action, and toxicokinetic characteristics of each of the 668 chemical substances appearing in the regulation. Each substance was assigned to one or more of 32 classes of biological effects retained by a group of toxicologists. The resulting database allows the user to find if there is potential additivity among components of a mixture.

The effect of rifampicin, a prototypical CYP3A4 inducer, on erlotinib pharmacokinetics in healthy subjects.

PURPOSE: Erlotinib, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy) quinazolin-4-amine is approved for the treatment for non-small cell lung cancer and pancreatic cancer. Because erlotinib is metabolized predominately by CYP3A4, co-administration of compounds that increase CYP3A4 activity may alter the efficacy and safety of erlotinib therapy. Two phase I studies were conducted in healthy male subjects to evaluate the effect of pre- or co-administered rifampicin, a CYP3A4 inducer, on the pharmacokinetics of erlotinib. METHODS: Study 1 included Groups A (erlotinib 150 mg days 1 and 15, rifampicin 600 mg days 8-14) and B (erlotinib 150 mg days 1 and 15) in a parallel group study design. Study 2 subjects received erlotinib 150 mg day 1, erlotinib 450 mg day 15, and rifampicin 600 mg days 8-18. The primary endpoint in each study was the ratio of exposure (AUC0-∞ and C max) between days 1 and 15. Urinary cortisol metabolic induction ratios were determined in Study 1 for Group A subjects only. RESULTS: In Study 1, the geometric mean ratios of AUC0-∞ and C max were 33 and 71 %, respectively, and the mean cortisol metabolic index increased from 7.4 to 27.0, suggesting cytochrome P450 (CYP) enzyme induction. In Study 2, the geometric mean ratios for AUC0-∞ and C max were 19 and 34 % (when dose adjusted from 450 to 150 mg erlotinib), respectively, a greater relative decrease than observed in Study 1. CONCLUSIONS: Erlotinib exposure (AUC0-∞ and C max) was reduced after pre- or concomitant dosing with rifampicin. Doses of ≥450 mg erlotinib may be necessary to compensate for concomitant medications with strong CYP3A4 enzyme induction effect.