Animal Utilization in Drug Development: Clinical, Legal,
7.10 ANIMAL ALTERNATIVES IN DRUG DEVELOPMENT
REPLACING, REDUCING, AND REFINEMENT OF ANIMALS Laboratory animal husbandry has been around since 1959, when the ap- proach to experimental techniques involving animal handling was revis- ited, requiring more caution and a humane approach to the use of animals.
Major Charles Hume, Founder of the Universities Federation for Animal Welfare, requested a publication on the application of ethical conduct in the use of animals in laboratory experiments. This was followed by two great scientists of the time: William Russell, a zoologist, psychologist, and classical scholar, and Rex Burch, a microbiologist, who subsequently published the 1959 Principles of Humane Experimental Techniques, which emphasized the principles of replacement, reduction, and refinement (the 3Rs) that are actively applied globally today.
7.10.1 Replacement
The depleted ecology, which includes animals that have been completely lost and extinct, is a possible impact of use of animals. Replacement alterna- tives require the use of other options where possible to minimize the use of animals. These involve the use of tissue cultures including embryonic and adult stem cells, and in vitro assays using mammalian or human cell cultures for a wide range of toxic and other endpoints. Modern cell culture techniques have been greatly improved due to emergent models that more closely mimic the in vivo milieu as explained in Chapter 5.
Microarray technology allows the assessment of large numbers of genes simultaneously, to permit expression profiling of genes that are activated or suppressed in response to examined molecular pathways or biological activi- ties. These are probed on a chip instead of the animal. Scientists and engineers at Harvard University have created “organs-on-a-chip,” including the “lung- on-a-chip” and “gut-on-a-chip.” These tiny devices contain human cells in a three-dimensional system that mimics human organs. Structure–activity relationships have enabled prediction of biological activity (e.g., toxicity) of drug compounds, which is based on the molecular structure and other in silico modeling activities. These strategies enable a “replacement” of animals as they are highly effective in disease research, drug testing, and toxicity testing.
Social Aspects of Drug Discovery, Development and Commercialization 164
In silico modeling is a computerized platform that utilizes known rules for predicting the related biological outcomes like metabolic fate, and these rules are mostly about factors affecting toxicity, physicochemical character- istics, and more. Typically, a range of chemical compounds undergo high- throughput screening (HTS) for a drug target or disease model prior to finding the preliminary “hits,” or active and functional motifs. These are involved in recognition and targeting activities that are well differentiated from other functional properties of the molecule in which they occur for the generation of cluster, potency, and factors like ligand efficiency or pre- defined toxicological factors.
7.10.1.1 Virtual Animals and Human Models
Computerized models of complex human biology attempt to reduce ani- mal testing and increase the speed of the drug discovery process. For ex- ample, a computerized model, PhysioLab®, is a product of a Californian biotechnology company Entelos. It is a precise algorithm, platform model with equations that describe the biological pathways that are adjusted to the existing information or client proprietary data. This is subsequently adapted to particular biological processes or pathways of interest. Its utility has been reported as an aid to target and biomarker selection, species translation, pa- tient stratification, and clinical trial design. The PhysioLab model has been applied to type 2 diabetes, rheumatoid arthritis, hypertension, and cardio- vascular disease modeling (http://www.entelos.com/). The computerized models might present their own pitfalls as no single human model exists due to human genetic diversity. Also, these models are based on certain gen- eralizations based on the most dominant genotypes and phenotypes. This might not be applicable in certain real-time cases and thus might not have a widespread application.
These valued fast-track technology-based accomplishments are the rea- son for modern-day HTS candidate drug screening, which takes place in a test tube and is subject to automation – a microscale representation of a human disease process. This understanding has been disputed based on the notion that a simplified model is not a true representative of complex human biology and its networks.
7.10.2 Reduction
Animal reduction strategies have been performed through improvement in experimental design and statistical analysis, compound prescreening (HTS), smaller focused studies, pilot tests that enable to properly calculate sample
sizes, improvement in education and staff training, policies of recycling of animals, and interim kills. The extent of invasiveness versus potential benefit derived from animal usage is taken into consideration prior to the decision on the number of animal subjects utilized in laboratory experiments. Cer- tain ethical, scientific, and practical considerations enhance the feasibility and predictivity of the animal model chosen.
7.10.3 Refinement
Refinement alternatives refine animal care, focusing on procedures that minimize pain and distress through the use of noninvasive imaging, oth- er convenient devices to avoid surgical trauma, and better blood sampling techniques like venipuncture. Positive reinforcement techniques have been widely applied, especially in primates.
The animal replacement strategies have not yet been entirely possible particularly due to the inadequacy of in silico technologies. However, nu- merous animal reduction strategies have led to clinical success.
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