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Systems biology approach for evaluating vaccine and adjuvant safety during preclinical and lot release testing
Abstracts / Toxicology Letters 280S (2017) S221–S227 1 Department of Environmental Health Sciences, Faculty of Pharmacy, Medical University of Warsaw, Warsaw, Poland 2 Department of Drug Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Warsaw, Poland
Number of patients diagnosed with depression is increasing worldwide, causing rise in antidepressant consumption. These pharmaceuticals are often taken chronically and their levels in patient plasma and tissue are high. Common problem is the simultaneously use of antidepressants with anti-inflammatory drugs and exposure of a patient skin to the light. Sunlight and solarium lights are the important abiotic factor influencing decomposition of chemical compounds. The aim of the research was assessment of diclofenac effect on antidepressants (sertraline, paroxetine and fluoxetine) photodegradation by using the bioassay and physicochemical methods. Spirostomum ambiguum was used for evaluation of cytotoxicity of the samples before and after irradiation in SunTestCPS+ solar simulator. Kinetic of photochemical degradation was analyzed by HPLC with PDA, while HPLC MSIMS was used for identification of photoproducts. The tested antidepressants were toxic towards S. ambiguum, while diclofenac was not toxic. Fluoxetine was stabile under exposure to light. The concentration of paroxetine decreased to 75% of the initial level during 3 h of exposure. Only in case of sertraline, the presence of diclofenac caused an indirect photodegradation. Sertraline concentration decreased by 30% and significant number of photoproducts was formed. The photoproducts had no impact to toxicity. Although initial results of this study showed that irradiation of tested pharmaceuticals did not led to the formation of more toxic photoproducts, comprehensive evaluation of phototoxicity of pharmaceuticals and their mixtures should be continued. Research was financed by the Medical University of Warsaw, Faculty of Pharmacy from the Grant for Young Scientists managed by Milena Wawryniuk (FW14IPM1I16). http://dx.doi.org/10.1016/j.toxlet.2017.07.614 P-07-03-07 Determination of exposure limits for risk analysis in shared facilities according to toxicological criteria: Implication of PDE calculation Beatriz Carrero 1 , Sabine Paris 2 1 2
Azierta, Madrid, Spain Maas & Peither GMP Verlag, Schopfheim, Germany
European Medicines Agency published (November 2014) the Guideline to define the limits of exposure to establish the risk in multiproduct facilities. It involves the determination of exposure limits according to toxicological criteria based on the inherent characteristics of each substance, instead of the traditional limits no-scientifically based. The procedure proposed in this document for determination of health based exposure limits is based on the method for establishing the Permitted Daily Exposure (PDE). In order to assist the pharmaceutical industry in implementing the new regulation, Azierta, as an external service provider, started a new project to calculate the PDE values for 1200 active pharmaceutical ingredients (APIs). After the toxicological evaluation, Azierta performed a thorough analysis of the obtained data. This huge and unique database allows a well-grounded revision and a close analysis of the results obtained. Categorization of PDE values was made, distinguishing 5 groups and assigning a level of dan-
S223
ger to each of them. The results were analysed according to the therapeutic groups (ATC classification). The analysis revealed that specific toxicological and pharmacological properties of each API has to be evaluated in order to establish health based exposure limits for medicinal products that are manufactured in shared facilities. It is not possible to estimate the danger or toxicity of APIs only by their therapeutic group. Not all substances with a high or very high toxicity level correspond to expected groups such as hormones and cytotoxic agents and, not all APIs in these therapeutic groups are highly hazardous. http://dx.doi.org/10.1016/j.toxlet.2017.07.615 P-07-03-08 Systems biology approach for evaluating vaccine and adjuvant safety during preclinical and lot release testing Takuo Mizukami 1 , Eita Sasaki 1 , Haruka Momose 1 , Yuki Hiradate 1 , Keiko Furuhata 1 , Hideki Kusunoki 1 , Hideki Asanuma 2 , Hiroshi Yamada 3 , Ken J. Ishii 4,5 , Isao Hamaguchi 1 1
Department of Safety Research on Blood and Biologicals, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo, Japan 2 Influenza Research Center, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo, Japan 3 Laboratory of Toxicogenomics Informatics, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan 4 Laboratory of Adjuvant Innovation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan 5 Laboratory of Vaccine Science, Immunology Frontier Research Center (IFREC), Osaka University, Osaka, Japan Vaccines are highly effective at preventing infectious diseases. Many vaccine components are derived from pathogens; therefore, ensuring their safety and consistency on a lot-to-lot basis is vital in both the preclinical phase and after licensure. The vaccine development platform has been modernized by introducing recombinant protein technology and new adjuvants that improve vaccine immunogenicity. However, the assessment methods used for vaccine and adjuvant safety have changed very little over time. Here, we show that systems biology approaches can evaluate vaccine and adjuvant safety. We focused on the influenza vaccine as it is the most sophisticated vaccine development platform. Laboratory rats were vaccinated with the influenza HA vaccine (HAV), or the whole particle influenza vaccine (WPV) as a toxicity reference. The global gene expression pattern in the rats revealed that the lung gene signatures clearly differentiated each vaccine. We selected 18 genes as vaccine biomarkers and assessed the quality of the seasonal HAV from several manufacturers. We found that our biomarkers could evaluate HAV quality with high sensitivity, whereas the conventional animal tests could not. We next assessed intranasal HAV with recently developed adjuvant such as CpG-K3, and found that these adjuvants and vaccine combination induced biomarker expression in a dose-dependent manner. We quantified gene expression level in the HAV-immunized mice and estimated the safety level compared with that of the WPV-immunized mice and found that the adjuvants tested are potentially safer than WPV. This approach has strong potential for evaluating vaccine and adjuvant safety. http://dx.doi.org/10.1016/j.toxlet.2017.07.616
Number of patients diagnosed with depression is increasing worldwide, causing rise in antidepressant consumption. These pharmaceuticals are often taken chronically and their levels in patient plasma and tissue are high. Common problem is the simultaneously use of antidepressants with anti-inflammatory drugs and exposure of a patient skin to the light. Sunlight and solarium lights are the important abiotic factor influencing decomposition of chemical compounds. The aim of the research was assessment of diclofenac effect on antidepressants (sertraline, paroxetine and fluoxetine) photodegradation by using the bioassay and physicochemical methods. Spirostomum ambiguum was used for evaluation of cytotoxicity of the samples before and after irradiation in SunTestCPS+ solar simulator. Kinetic of photochemical degradation was analyzed by HPLC with PDA, while HPLC MSIMS was used for identification of photoproducts. The tested antidepressants were toxic towards S. ambiguum, while diclofenac was not toxic. Fluoxetine was stabile under exposure to light. The concentration of paroxetine decreased to 75% of the initial level during 3 h of exposure. Only in case of sertraline, the presence of diclofenac caused an indirect photodegradation. Sertraline concentration decreased by 30% and significant number of photoproducts was formed. The photoproducts had no impact to toxicity. Although initial results of this study showed that irradiation of tested pharmaceuticals did not led to the formation of more toxic photoproducts, comprehensive evaluation of phototoxicity of pharmaceuticals and their mixtures should be continued. Research was financed by the Medical University of Warsaw, Faculty of Pharmacy from the Grant for Young Scientists managed by Milena Wawryniuk (FW14IPM1I16). http://dx.doi.org/10.1016/j.toxlet.2017.07.614 P-07-03-07 Determination of exposure limits for risk analysis in shared facilities according to toxicological criteria: Implication of PDE calculation Beatriz Carrero 1 , Sabine Paris 2 1 2
Azierta, Madrid, Spain Maas & Peither GMP Verlag, Schopfheim, Germany
European Medicines Agency published (November 2014) the Guideline to define the limits of exposure to establish the risk in multiproduct facilities. It involves the determination of exposure limits according to toxicological criteria based on the inherent characteristics of each substance, instead of the traditional limits no-scientifically based. The procedure proposed in this document for determination of health based exposure limits is based on the method for establishing the Permitted Daily Exposure (PDE). In order to assist the pharmaceutical industry in implementing the new regulation, Azierta, as an external service provider, started a new project to calculate the PDE values for 1200 active pharmaceutical ingredients (APIs). After the toxicological evaluation, Azierta performed a thorough analysis of the obtained data. This huge and unique database allows a well-grounded revision and a close analysis of the results obtained. Categorization of PDE values was made, distinguishing 5 groups and assigning a level of dan-
S223
ger to each of them. The results were analysed according to the therapeutic groups (ATC classification). The analysis revealed that specific toxicological and pharmacological properties of each API has to be evaluated in order to establish health based exposure limits for medicinal products that are manufactured in shared facilities. It is not possible to estimate the danger or toxicity of APIs only by their therapeutic group. Not all substances with a high or very high toxicity level correspond to expected groups such as hormones and cytotoxic agents and, not all APIs in these therapeutic groups are highly hazardous. http://dx.doi.org/10.1016/j.toxlet.2017.07.615 P-07-03-08 Systems biology approach for evaluating vaccine and adjuvant safety during preclinical and lot release testing Takuo Mizukami 1 , Eita Sasaki 1 , Haruka Momose 1 , Yuki Hiradate 1 , Keiko Furuhata 1 , Hideki Kusunoki 1 , Hideki Asanuma 2 , Hiroshi Yamada 3 , Ken J. Ishii 4,5 , Isao Hamaguchi 1 1
Department of Safety Research on Blood and Biologicals, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo, Japan 2 Influenza Research Center, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo, Japan 3 Laboratory of Toxicogenomics Informatics, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan 4 Laboratory of Adjuvant Innovation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan 5 Laboratory of Vaccine Science, Immunology Frontier Research Center (IFREC), Osaka University, Osaka, Japan Vaccines are highly effective at preventing infectious diseases. Many vaccine components are derived from pathogens; therefore, ensuring their safety and consistency on a lot-to-lot basis is vital in both the preclinical phase and after licensure. The vaccine development platform has been modernized by introducing recombinant protein technology and new adjuvants that improve vaccine immunogenicity. However, the assessment methods used for vaccine and adjuvant safety have changed very little over time. Here, we show that systems biology approaches can evaluate vaccine and adjuvant safety. We focused on the influenza vaccine as it is the most sophisticated vaccine development platform. Laboratory rats were vaccinated with the influenza HA vaccine (HAV), or the whole particle influenza vaccine (WPV) as a toxicity reference. The global gene expression pattern in the rats revealed that the lung gene signatures clearly differentiated each vaccine. We selected 18 genes as vaccine biomarkers and assessed the quality of the seasonal HAV from several manufacturers. We found that our biomarkers could evaluate HAV quality with high sensitivity, whereas the conventional animal tests could not. We next assessed intranasal HAV with recently developed adjuvant such as CpG-K3, and found that these adjuvants and vaccine combination induced biomarker expression in a dose-dependent manner. We quantified gene expression level in the HAV-immunized mice and estimated the safety level compared with that of the WPV-immunized mice and found that the adjuvants tested are potentially safer than WPV. This approach has strong potential for evaluating vaccine and adjuvant safety. http://dx.doi.org/10.1016/j.toxlet.2017.07.616