Research on the development of alternatives to animal testing for pharmaceutical products and international standardisation

Impact ◽  
2021 ◽  
Vol 2021 (8) ◽  
pp. 44-45
Author(s):  
Hajime Kojima
Keyword(s):  
Research Group ◽  
Pharmaceutical Products ◽  
In Vitro Assays ◽  
Animal Testing ◽  
3 Dimensional ◽  
Alternatives To Animal Testing ◽  
Gene Assay ◽  
Innovative Work ◽  
Regulatory Acceptance

Scientists are working to develop new and innovative alternatives to animal testing that don't rely on the use of animals. Takao Ashikaga, Hajime Kojima and Yoko Hirabayashi are part of JaCVAM which works to promote the use of alternatives to animal testing. The goal is to replace, reduce or refine (3Rs) the use of animal under International harmonization. Hirabayashi is also the representative of a research group that is funded by the AMED and the representative of a research group funded by the MHLW. A challenge the researchers are facing in their quest to ensure the welfare of experimental animals and also ensure the safety of various pharmaceutical and chemicals is the lack of biomarkers to more accurately predict toxicity for regulatory acceptance. This means that without animal testing more costly and complex non-animal methods are required and presents a barrier to the adoption of non-animal methods for international standerisation. As such, there is a need to develop an easy way to obtain a lot of information. Hirabayashi and the team are working on the development of AI that can be used to evaluate the safety of different compounds. The researchers are developing in vitro assays such as ordinary 2-dimensional culture, 3-dimensional culture including organoids or spheroids, reporter gene assay and organ-on-a chip; and in silico assays such as computer toxicology using QSAR and Read Across. The researchers hope that their innovative work will contribute to the 3Rs, benefiting animal welfare for regulatory use.

scholarly journals A Next-Generation Risk Assessment Case Study for Coumarin in Cosmetic Products

2020 ◽  
Vol 176 (1) ◽  
pp. 236-252 ◽  
Author(s):  
Maria T Baltazar ◽  
Sophie Cable ◽  
Paul L Carmichael ◽  
Richard Cubberley ◽  
Tom Cull ◽  
...  
Keyword(s):  
Risk Assessment ◽  
Biological Effects ◽  
In Vitro Assays ◽  
Next Generation ◽  
Animal Testing ◽  
Immunomodulatory Effects ◽  
Margin Of Safety

Abstract Next-Generation Risk Assessment is defined as an exposure-led, hypothesis-driven risk assessment approach that integrates new approach methodologies (NAMs) to assure safety without the use of animal testing. These principles were applied to a hypothetical safety assessment of 0.1% coumarin in face cream and body lotion. For the purpose of evaluating the use of NAMs, existing animal and human data on coumarin were excluded. Internal concentrations (plasma Cmax) were estimated using a physiologically based kinetic model for dermally applied coumarin. Systemic toxicity was assessed using a battery of in vitro NAMs to identify points of departure (PoDs) for a variety of biological effects such as receptor-mediated and immunomodulatory effects (Eurofins SafetyScreen44 and BioMap Diversity 8 Panel, respectively), and general bioactivity (ToxCast data, an in vitro cell stress panel and high-throughput transcriptomics). In addition, in silico alerts for genotoxicity were followed up with the ToxTracker tool. The PoDs from the in vitro assays were plotted against the calculated in vivo exposure to calculate a margin of safety with associated uncertainty. The predicted Cmax values for face cream and body lotion were lower than all PoDs with margin of safety higher than 100. Furthermore, coumarin was not genotoxic, did not bind to any of the 44 receptors tested and did not show any immunomodulatory effects at consumer-relevant exposures. In conclusion, this case study demonstrated the value of integrating exposure science, computational modeling and in vitro bioactivity data, to reach a safety decision without animal data.

Dialogue and Collaboration: A Personal View on Laboratory Animal Welfare Developments in General, and on ECVAM's First Decade in Particular

2002 ◽  
Vol 30 (2_suppl) ◽  
pp. 207-210 ◽  
Author(s):  
Herman B.W.M. Koëter
Keyword(s):  
National Level ◽  
In Vitro Tests ◽  
Practical Application ◽  
Animal Testing ◽  
Validation Process ◽  
Personal View ◽  
Ld50 Test ◽  
The Three Rs ◽  
Regulatory Acceptance

A personal view is presented on progress made during the last 25 years in applying the Three Rs ( reduction, refinement, replacement) to animal testing in regulatory toxicology, with an emphasis on “good moments” (for example, international workshops on the principles and practical application of the validation process and on regulatory acceptance) and “not-so-good moments” (for example, the time taken to accept alternatives to the LD50 test and to accept in vitro tests for skin absorption as OECD Test Guidelines). The importance of dialogue and cooperation between international coordinating centres and scientific activities at the national level is stressed, as exemplified by the work of ECVAM during its first decade.

scholarly journals Biological Activities of Novel Gyrase Inhibitors of the Aminocoumarin Class

2008 ◽  
Vol 52 (6) ◽  
pp. 1982-1990 ◽  
Author(s):  
Christine Anderle ◽  
Martin Stieger ◽  
Matthew Burrell ◽  
Stefan Reinelt ◽  
Anthony Maxwell ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Rank Order ◽  
Biological Activities ◽  
Dna Supercoiling ◽  
Binding Energies ◽  
In Vitro Assays ◽  
Gene Assay ◽  
Gyrase Inhibitors

ABSTRACT Thirty-one aminocoumarin antibiotics derived from mutasynthesis experiments were investigated for their biological activities. Their inhibitory activities toward Escherichia coli DNA gyrase were determined in two different in vitro assays: an ATPase assay and a DNA supercoiling assay. The assays gave a similar rank order of the activities of the compounds tested, although the absolute 50% inhibitory concentrations (IC50s) obtained in each assay were different. To confirm that the compounds also acted as gyrase inhibitors in vivo, reporter gene assays were carried out with E. coli by using gyrA and sulA promoter fusions with the luxCDABE operon. A strong induction of both promoters was observed for those compounds that showed gyrase inhibitory activity in the biochemical assays. Compounds carrying analogs of the prenylated benzoyl moiety (ring A) of clorobiocin that were structurally very different showed high levels of activity both in the biochemical assay and in the reporter gene assay, indicating that the structure of this moiety can be varied considerably without a loss of affinity for bacterial gyrase. The experimentally determined IC50s were compared to the binding energies calculated in silico, which indicated that a shift of the pyrrole carboxylic acid moiety from the O-3″ to the O-2″ position of the deoxysugar moiety has a significant impact on the binding mode of the compounds. The aminocoumarin compounds were also investigated for their MICs against different bacterial pathogens. Several compounds showed high levels of activity against staphylococci, including a methicillin-resistant Staphylococcus aureus strain. However, they showed only poor activities against gram-negative strains.

Canonical NF-kB Pathway Is Not Constitutively Activated and May Not Be the Major Bortezomib Target in Mantle Cell Lymphoma (MCL).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2591-2591
Author(s):  
Valerie Camara-Clayette ◽  
Yves Lepelletier ◽  
Ivan Moura ◽  
William Vainchenker ◽  
Olivier Hermine ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
Cell Lines ◽  
Myeloma Cell ◽  
Dominant Negative ◽  
In Vitro Assays ◽  
Lymphoid Malignancies ◽  
Empty Vector ◽  
Gene Assay

Abstract Bortezomib, a proteasome inhibitor, has promising activity in MCL. Bortezomib mechanism of action is complex since multiple proteins and cellular pathways are regulated by proteasome degradation. By blocking the degradation of IKBa, the canonical NF-kB pathway is one of the major Bortezomib target in lymphoid malignancies such as multiple myeloma. NF-kB activation has not been extensively assessed in other lymphoid malignancies such as MCL. NF-kB complexes (heterodimers P50/P65) were assessed by EMSA assays in REC GRANTA 519, NCEB, JEKO, JUN and UPN1 MCL cell-lines. EBV was present in 3/6 cell lines. P50/P65 complexes were detected in all EBV positive cell lines and only in 1/3 EBV negative cell-lines (JEKO). No nuclear P65 protein was detected by immunofluorescence or Western-blot analysis in the 2 other negative EBV cell lines (REC and UPN1). NF-kB transcriptional activity was measured by a luciferase-based reporter gene assay. Spontaneous NF-kB activity was low in UPN1, REC and JEKO compared to GRANTA 519 EBV positive cell-line (3 to 4 time lower) but can be strongly activated (up to 10 fold) using MEKK cotransfection assay. This suggest that NF-kB is functional, but not constitutively activated. Stable infections with Migr1-IRES-GFP IkBM (dominant negative IKB) or empty vector, were performed in GRANTA 519 EBV positive as well as in UPN1 cell line. No differences in proliferation or apoptosis were observed in UPN1 stably infected with IkBM or with the empty vector but GRANTA 519 EBV positive showed increased apoptosis and proliferation inhibition when infected with IkBM. Likewise, no P50/P65 heterodimers complexes were detected by EMSA in 4 patients with MCL. In vitro assays showed that UPN1 and JEKO cell lines had comparable sensitivities to Bortezomib than Multiple myeloma cell lines reported in the literature (IC 50: 6 nM and 12 nM respectively). This was also true for the 3 patients lymphoma cells assessed in vitro (12 nM). Taken together these results suggest that canonical NF-kB activation pathway is not constitutively active in EBV negative MCL cell lines and patients samples and could be associated with EBV infection in some MCL cell-lines. Therefore, this strongly suggests that Bortezomib target other molecules in MCL.

In Vitro Methods: Alternatives to Animal Testing

2019 ◽  
pp. 447-457
Author(s):  
Dayane Pifer Luco ◽  
Vânia Rodrigues Leite-Silva ◽  
Heather A.E. Benson ◽  
Patricia Santos Lopes
Keyword(s):  
Animal Testing ◽  
In Vitro Methods ◽  
Alternatives To Animal Testing

scholarly journals The use of in vitro assays for the assessment of cytotoxicity on the example of MTT test

2018 ◽  
Vol 14 ◽  
pp. 23-32 ◽  
Author(s):  
Magdalena Twarużek ◽  
Ewa Zastempowska ◽  
Ewelina Soszczyńska ◽  
Iwona Ałtyn
Keyword(s):  
In Vitro Assays ◽  
Toxic Substances ◽  
Animal Testing ◽  
Qualitative Information ◽  
Chemical Methods ◽  
Mtt Test ◽  
Bacterial Cultures ◽  
Biological Tests ◽  
Bio Assay

In recent years, biological tests have been developed based on cell cultures and successfully used to the hygienic assess of a variety of samples. In vitro assays become the complement of conventional chemical methods. They do not narrow the results only to the quantitative and qualitative information on toxic substances, but also increase knowledge on the direct impact on the organism. They are also an alternative for animal testing, which are currently given up for ethical reasons. At present, the market is steadily increasing in the number of tests and bio-assay techniques. Based on our own studies we conclude that the MTT test is perfect as a diagnostic method for evaluating the cytotoxicity of materials of different composition such as mycotoxins, pesticides, bacterial cultures, moulds isolates, food, feed, as well as a vast spectrum of other environmental samples.

2018 Lush Science Prize

2020 ◽  
Vol 48 (1_suppl) ◽  
pp. 18S-25S
Author(s):  
Jenny McCann ◽  
Terry McCann
Keyword(s):  
Public Awareness ◽  
Relevant Literature ◽  
Toxicity Testing ◽  
In Vitro Assays ◽  
Adverse Outcome Pathway ◽  
Annual Conference ◽  
Animal Testing ◽  
Safety Testing ◽  
Animal Replacement

The Lush Prize supports animal-free testing by awarding money prizes of up to £350,000 per year to the most effective projects and individuals who have been working towards the goal of replacing animals in product or ingredient safety testing. Since its inception in 2012, the Lush Prize has distributed almost £2 million. Prizes are awarded for developments in five strategic areas: Science; Lobbying; Training; Public Awareness; and Young Researchers. In 2015, the judges also awarded a Black Box prize for the development of the skin sensitisation Adverse Outcome Pathway and its associated in vitro assays. The Science Prize is awarded to researchers whose work the judging panel believe to have made the most significant contribution, in the preceding year, to the replacement of animal testing. This 2018 Science Background paper outlines the research projects that were presented to the Prize judges as potential candidates for the 2018 Lush Science Prize award. To obtain an overview of developments in the field of animal replacement in toxicity research, recent work by the relevant scientific institutions and projects in this area, including the OECD, CAAT, ECVAM, UK NC3Rs, US Tox21 Programme, the ToxCast programme and EU-ToxRisk, was reviewed. Recent developments in toxicity testing research were investigated by searching the relevant literature. Abstracts from conferences focusing on animal replacement in toxicity testing that were held in the preceding 12 months, were also analysed, including those from the 2017 10th World Congress on Alternatives and Animals in the Life Sciences and the 2018 Society of Toxicology annual conference.

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