Jürg Oliver Straub

A strategy to reduce the numbers of fish used in acute ecotoxicity testing of pharmaceuticals

The pharmaceutical industry gives high priority to animal welfare in the process of drug discovery and safety assessment. In the context of environmental assessments of active pharmaceutical ingredients (APIs), existing U.S. Food and Drug Administration and draft European regulations may require testing of APIs for acute ecotoxicity to algae, daphnids, and fish (base-set ecotoxicity data used to derive the predicted no-effect concentration [PNECwater] from the most sensitive of three species). Subject to regulatory approval, it is proposed that testing can be moved from fish median lethal concentration (LC50) testing (typically using > or = 42 fish/API) to acute threshold tests using fewer fish (typically 10 fish/API). To support this strategy, we have collated base-set ecotoxicity data from regulatory studies of 91 APIs (names coded for commercial reasons). For 73 of the 91 APIs, the algal median effect concentration (EC50) and daphnid EC50 values were lower than or equal to the fish LC50 data. Thus, for approximately 80% of these APIs, algal and daphnid acute EC50 data could have been used in the absence of fish LC50 data to derive PNECwater values. For the other 18 APIs, use of an acute threshold test with a step-down factor of 3.2 is predicted to give comparable PNECwater outcomes. Based on this preliminary scenario of 91 APIs, this approach is predicted to reduce the total number of fish used from 3,822 to 1,025 (approximately 73%). The present study, although preliminary, suggests that the current regulatory requirement for fish LC50 data regarding APIs should be succeeded by fish acute threshold (step-down) test data, thereby achieving significant animal welfare benefits with no loss of data for PNECwater estimates.

Environmental risk assessment for new human pharmaceuticals in the European Union according to the draft guideline/discussion paper of January 2001

Since 1993, an environmental risk assessment (ERA) for a new drug application has been stipulated by EU Directive 93/39/EEC amending Directive 65/65/EEC. In early 2001, after several unpublished draft versions for an ERA guideline, a draft guideline/discussion paper for an ERA for non-GMO-containing drugs was published by the European Medicines Evaluation Agency (EMEA). The draft guideline describes a step-wise, tiered procedure for the ERA. The first tier consists of deriving a crude predicted environmental concentration (PEC) in the aquatic compartment for the active pharmaceutical ingredient (API) or its major metabolites, based on predicted amounts used and specific removal rates in sewage treatment or surface waters. If this crude PEC is <0.01 microg/l and no environmental concerns are apparent, no further assessment is deemed necessary. Else, in the second tier, a crude predicted no-effect level (PNEC) for the aquatic compartment is to be extrapolated by dividing the lowest 50%-effect concentration from acute ecotoxicity tests with algae, daphnia or fish (EC(50), LC(50)) by an assessment factor (usually 1000). If the ratio PEC/PNEC is <1, no further assessment is deemed necessary. Lastly, in the third tier, further considerations on a case-by-case basis are needed. This may encompass refining the environmental fate information and thereby the PEC, considering further environmental compartments and their respective PECs (up to and including field studies), but also refining the PNEC. While the ERA addresses mainly the API, excipients of the formulated drug should be considered as well. In the case of medicinal products, the benefit for patients has relative precedence over environmental risks, meaning that even in the case of an unacceptable residual risk for new drugs after third-tier considerations, prohibition of a new API is not taken into consideration. Instead, possible mitigating or precautionary safety measures may consist of specific product labelling (i.e. package leaflets for the patients regarding returning and proper disposal of unused medicines), restricted use through in-hospital or in-surgery administration under supervision only, or the recommendation of environmental analytical monitoring up to ecological field studies.

An environmental risk assessment for oseltamivir (Tamiflu®) for sewage works and surface waters under seasonal-influenza- and pandemic-use conditions.

In the event of an influenza pandemic, anti-viral medications such as oseltamivir (Tamiflu) are expected to be used in high amounts over a duration of several weeks. Oseltamivir has been predicted to reach high concentrations in surface waters and sewage works. New oseltamivir environmental fate and toxicity studies permit an environmental risk assessment (ERA) under seasonal- and pandemic-use scenarios. The environmental fate data for sewage works (no removal), surface waters (no significant degradation), and water/sediment systems (>50% primary degradation in 100 days) were used for the derivation of new predicted environmental concentrations (PECs) for western Europe and the River Lee catchment in the UK. Existing worst-case PECs for western Europe, the River Lee catchment in the UK and the Lower Colorado basin in the USA under pandemic conditions (< or =98.1 microg/L for surface waters, < or =348 microg/L for sewage works) were also considered for the ERA. PECs were compared with predicted no-effect concentrations (PNECs) based on new chronic ecotoxicity data (no observed effect concentration for algae, daphnia, and fish > or =1 mg/L). Based on all PEC/PNEC risk ratios, no significant risk is evident to surface waters or sewage works during both regular seasonal-use and high pandemic-use of oseltamivir.

Meeting Report: Risk Assessment of Tamiflu Use Under Pandemic Conditions

On 3 October 2007, 40 participants with diverse expertise attended the workshop Tamiflu and the Environment: Implications of Use under Pandemic Conditions to assess the potential human health impact and environmental hazards associated with use of Tamiflu during an influenza pandemic. Based on the identification and risk-ranking of knowledge gaps, the consensus was that oseltamivir ethylester-phosphate (OE-P) and oseltamivir carboxylate (OC) were unlikely to pose an ecotoxicologic hazard to freshwater organisms. OC in river water might hasten the generation of OC-resistance in wildfowl, but this possibility seems less likely than the potential disruption that could be posed by OC and other pharmaceuticals to the operation of sewage treatment plants. The work-group members agreed on the following research priorities: a) available data on the ecotoxicology of OE-P and OC should be published; b) risk should be assessed for OC-contaminated river water generating OC-resistant viruses in wildfowl; c) sewage treatment plant functioning due to microbial inhibition by neuraminidase inhibitors and other antimicrobials used during a pandemic should be investigated; and d) realistic worst-case exposure scenarios should be developed. Additional modeling would be useful to identify localized areas within river catchments that might be prone to high pharmaceutical concentrations in sewage treatment plant effluent. Ongoing seasonal use of Tamiflu in Japan offers opportunities for researchers to assess how much OC enters and persists in the aquatic environment.

Combined environmental risk assessment for 5‐fluorouracil and capecitabine in Europe

An environmental risk assessment (ERA) was made for the old cytostatic active pharmaceutical ingredient 5-fluorouracil (5-FU) and for capecitabine (CAP), which is a prodrug of 5-FU. This ERA is based on published and company internal data as well as new test results for physicochemical, human metabolism, biodegradability, environmental partitioning and fate, and acute and chronic ecotoxicity properties of the active substance 5-FU as well as on use sales data for 5-FU and CAP in Europe. Predicted environmental concentrations (PECs) were extrapolated following the EMEA 2006 Guideline on ERA for human pharmaceuticals and the European Union 2003 Technical Guidance Document (TGD) for risk assessment as well as the TGD-based application EUSES v2.0. Actual amounts sold were taken from IMS Health Databases, in order to refine the default use and EMEA penetration factor as well as the PECs. Moreover, available measured environmental concentrations (MECs) were used to supplement PECs. A predicted no-effect concentration (PNEC) for 5-FU was derived from chronic ecotoxicity data. Except for the simplistic EMEA Phase I default PEC, the risk characterization by PEC:PNEC and MEC:PNEC ratios for various environmental compartments resulted in no significant risk. As the EMEA Phase I PEC does not integrate documented human metabolism and environmental degradation, in contrast to refined PEC derivations, it is inferred that the current use of CAP and 5-FU does not present any evident risk to the environment. An additional evaluation of persistence, bioaccumulation, and toxicity (PBT) properties supports the conclusion of no significant environmental risk for 5-FU and CAP.

Deterministic and probabilistic acute‐based environmental risk assessment for naproxen for Western Europe

An environmental risk assessment (ERA) was made for the common nonsteroidal anti-inflammatory drug naproxen. The ERA was performed according to deterministic and probabilistic methods, based on different predicted environmental concentrations (PECs) and measured environmental concentrations (MECs) on the exposure side as well as on published and newly elaborated acute ecotoxicity data on the effects side. Compilation of a large set of MECs allowed a qualification of the various PEC derivations. The European Medicines Evaluation Authority (EMEA) phase I PEC was shown to be far above realistic values, while the refined EMEA phase II (A and B) PECs were not too far from the 95th percentile MEC, in agreement with their nature as local PECs. The western European continental and regional PECs extrapolated based on actual use data, using the European Union system for the evaluation of substances, with the region reconfigured for Germany where most of the available European MECs are from, were in good to very close agreement with the median MECs. No risk to surface waters is apparent by any of the methodologies applied from the current use of naproxen; however, because only insufficient chronic ecotoxicity data are available, this is a preliminary conclusion.

Extending the environmental risk assessment for oseltamivir (Tamiflu ® ) under pandemic use conditions to the coastal marine compartment

In case of an avian-influenza-derived human flu pandemic, an inordinately high use of medicines over several weeks is predicted, in particular for the recommended influenza antiviral oseltamivir (Tamiflu). While the risk of oseltamivir to sewage works and freshwater bodies has already been assessed, the fact that a large percentage of the human population worldwide lives relatively close to the sea raises concern for its environmental compatibility in coastal marine waters. The potential risk of high oseltamivir use to the marine compartment is assessed in this publication, based on the 2003 European Community Technical Guidance Document (TGD) for risk assessment. Subchronic embryo-larval ecotoxicity tests with three marine invertebrates (Pomatoceros triqueter, Annelida; Mytilus edulis, Mollusca; Paracentrotus lividus, Echinodermata) and chronic growth inhibition tests with two different groups of marine microalgae (Isochrysis galbana, Haptophyta; Skeletonema costatum, Heterokontophyta) were performed with the active substance oseltamivir carboxylic acid to derive a dependable marine predicted no-effect concentration (PNEC). This was compared to a predicted environmental concentration (PEC) for oseltamivir in coastal waters, based on the worst-case freshwater PEC. The PEC/PNEC risk characterisation ratio for the marine compartment is well below 1, which in the terminology of the TGD signifies no immediate concern. Further, while oseltamivir may be persistent (P), it is not bioaccumulative (B) nor highly ecotoxic (T) and therefore not a PBT substance. In conclusion, even a high pandemic use of oseltamivir would not lead to a significant risk for the marine compartment, in confirmation of the risk assessment for sewage works and freshwaters.

Use of acute and chronic ecotoxicity data in environmental risk assessment of pharmaceuticals

For many older pharmaceuticals, chronic aquatic toxicity data are limited. To assess risk during development, scale-up, and manufacturing processes, acute data and physicochemical properties need to be leveraged to reduce potential long-term impacts to the environment. Aquatic toxicity data were pooled from daphnid, fish, and algae studies for 102 active pharmaceutical ingredients (APIs) to evaluate the relationship between predicted no-effect concentrations (PNECs) derived from acute and chronic tests. The relationships between acute and chronic aquatic toxicity and the n-octanol/water distribution coefficient (D(OW)) were also characterized. Statistically significant but weak correlations were observed between toxicity and log D(OW), indicating that D(OW) is not the only contributor to toxicity. Both acute and chronic PNEC values could be calculated for 60 of the 102 APIs. For most compounds, PNECs derived from acute data were lower than PNECs derived from chronic data, with the exception of steroid estrogens. Seven percent of the PNECs derived from acute data were below the European Union action limit of 0.01 μg/L and all were anti-infectives affecting algal species. Eight percent of available PNECs derived from chronic data were below the European Union action limit, and fish were the most sensitive species for all but 1 API. These analyses suggest that the use of acute data may be acceptable if chronic data are unavailable, unless specific mode of action concerns suggest otherwise.

Concentrations of the UV filter ethylhexyl methoxycinnamate in the aquatic compartment: a comparison of modelled concentrations for Swiss surface waters with empirical monitoring data.

UV filters in sunscreens and cosmetics protect the skin from damage through UV radiation. Many tonnes per year of UV filters are being used in Europe and will be present, at least seasonally, in detectable concentrations in surface waters similar to common pharmaceutically active substances. Predicted environmental concentrations (PECs) of ethylhexyl methoxycinnamate (EHMC; CAS 5466-77-3) were extrapolated for Switzerland, taking into consideration substance-specific environmental fate data and marketing estimates, by crude worst-case reckoning and by applying two environmental models (Mackay Level III; USES 3.0), both configured for Swiss hydrological and area data. By worst-case reckoning the summer PEC is 70.8-81.3 ng/l while for the remaining 8 months of the year the PEC is 13.1-15.1 ng/l. The Level III model results in concentrations of 2.4 ng/l during the summer and 0.44 ng/l during the rest of the year, while the USES 3.0 model gives an average PEC for the whole year of 7.6 ng/l. Pooling summer monitoring data (90 single analyses) from the River Rhine below Basel in the year 1997 (Water Protection Board of Basel) and from Lakes Zurich and Hüttner in 1998 (Poiger et al., in preparation) allowed a derivation of a probabilistic median concentration of 4.6 ng/l, a 95th-percentile concentration of 18.6 ng/l and a 99th-percentile concentration of 33.5 ng/l. The 6-fold range from the median value to the maximum calls for caution in interpreting published monitoring concentrations. Comparison of modelled PECs with realistic median concentrations shows that crude reckoning overestimates actual concentrations by a factor of about 10, probably through insufficient consideration of (further) degradation of EHMC in sewage works, surface waters, sediments or river banks. Both computer models, in contrast, are within the same order of magnitude as the actual summer concentrations. Based on the available data, both these environmental fate and distribution models give realistic PECs.

Performance of an anaerobic membrane bioreactor for pharmaceutical wastewater treatment

Anaerobic treatment of wastewater and waste organic solvents originating from the pharmaceutical and chemical industries was tested in a pilot anaerobic membrane bioreactor, which was operated for 580days under different operational conditions. The goal was to test the long-term treatment efficiency and identify inhibitory factors. The highest COD removal of up to 97% was observed when the influent concentration was increased by the addition of methanol (up to 25gL-1 as COD). Varying and generally lower COD removal efficiency (around 78%) was observed when the anaerobic membrane bioreactor was operated with incoming pharmaceutical wastewater as sole carbon source. The addition of waste organic solvents (>2.5gL-1 as COD) to the influent led to low COD removal efficiency or even to the breakdown of anaerobic digestion. Changes in the anaerobic population (e.g., proliferation of the genus Methanosarcina) resulting from the composition of influent were observed.