Science Pool

Antibodies generated in the lab are important as potential treatments for a broad spectrum of diseases, in particular infectious diseases caused by viruses. They can be obtained either by animal-derived B cells or from antibody library display platforms. Evotec’s strategy for the optimal path to obtain lead candidates is offering access to both sources of antibodies for discovery, coupled with the exploitation of state-of- the-art technologies to ensure success for a broad range of targets and disease states. In addition, selected lead candidates can be further optimized using powerful computational platforms to enhance productivity, manufacturability, and formulation stability. This is the end-to-end J.Design biologics platform, which is fueled by the front-end discovery platform, J.HAL™ (Just Humanoid Antibody Library) and associated data-driven, company-wide machine learning methodology.

By using artificial intelligence (AI) and machine learning (ML), J.HAL can generate novel, humanoid antibody sequences that both represent natural repertoires and are biased towards desirable features. To enable properties such as broad target and epitope engagement, focused efficacy, and suitable developability, Just-Evotec Biologics has devised an Antibody-GAN (Generative Adversarial Network), a new synthetic approach to designing a novel class of antibody therapeutics, which is termed humanoid antibodies.

At the conferences International Conference on Antiviral Research (ICAR) 2021 and Antibody Engineering & Therapeutics Europe 2022, researchers from Evotec and Just-Evotec Biologics introduced results obtained by using GAN to generate novel sequences, which mimic natural human response and provide the necessary diversity and developability features.

Competing Neural Networks

GAN is based on competing, deep layer neural networks that learn and produce the features of the mature human antibody repertoire, including sequence characteristics and structure properties, allowing for the encoding of key properties of interest into diverse libraries for a feature-biased discovery platform. It works to:

• capture the complexity of the entire variable region of the standard human antibody sequence space,
• provide a basis for generating novel antibodies that span a larger sequence diversity than standard in silico generative approaches, and
• incorporate transfer learning, a critical feature for antibody discovery to bias the physical properties of the generated antibodies towards broader efficacy traits such as CDR lengths and surface properties, improved developability (e.g., improved thermal and pH stability), and diverse chemical and biophysical properties.

The GAN network is trained by using hundreds of thousands of human antibody sequences to recognize legitimate human v-genes. The generator network generates random sequences to fool the discriminator while continually receiving feedback from the discriminator on sequence validity. Over time, the two networks get progressively better at their tasks. After full training, the Antibody-GAN generator is eventually able to produce fully human, novel antibody sequences for the germline for which the GAN was trained.

Antibodies targeting SARS-CoV-2

To demonstrate the usefulness of this platform, the researchers used their newly constructed, 1 billion theoretical diversity phage Fab library with the intent to discover antibodies to the SARS-CoV-2 spike protein. Candidates that specifically bound SARS-CoV-2 spike protein and did not bind an irrelevant antigen were further characterized for dose-dependent binding using AlphaLISA technology. In the primary “yes/no” binding screen a total of 73 unique antibody sequences specific for SARS-CoV-2 spike protein were identified. The researchers then performed binding assays using unpurified transfection supernatants and later reproduced the results with purified material. The candidate antibody supernatants that specifically bound SARS-CoV-2 spike protein were subsequently tested for their ability to block binding of this protein to human ACE-2 receptor. The team identified multiple antibodies that effectively blocked spike human ACE2 receptor interaction, demonstrating the feasibility to screen unpurified transfection supernatants for functional activity. After further rounds of panning, the top candidates expressed at flask scale were purified and tested for SARS-CoV-2 neutralization ability across multiple strains. The researchers identified multiple candidates with neutralizing activity against several strains of SARS-CoV-2. Nine of these antibodies exhibited blocking activity of the spike protein to the ACE2 receptor in an in vitro functional assay. Of note, all antibody data shown here were from native library candidates without any affinity maturation.

The presentation demonstrates that applying machine learning algorithms in antibody discovery “promotes efficient learning from the least expensive and most abundant data encoded in the DNA of antibodies, to validation of this learning through less abundant, more expensive, but most relevant data from GMP manufacturing at full commercial scale,” stated James N. Thomas, retired Executive Vice President, Global Head of Biotherapeutics and President U.S. Operations at Just - Evotec Biologics. “This is a systems approach to platform definition and continuous improvement, and it is unique in the industry, made possible by a number of factors that will be difficult for others to replicate."

To learn more about Evotec's capabilities read our related poster.

Learn More

iPSC and age-related diseases: Case study AMD
Age-related macular degeneration (AMD) is a leading cause of vision loss in people over the age of 50, accounting for 90% of blindness in this population. There are dry and wet forms of the disease and to date, there are limited treatment options for the wet form and none for dry AMD. The exact cause of the disease is unknown, but it is suspected that it results from a combination of hereditary and environmental factors, with smoking and diet being implicated. Globally, more than 190 million people are affected by AMD and this figure is expected to increase to more than 285 million by the year 2040. The estimated global cost of the disease currently stands at $343bn, with $255bn in direct healthcare costs.

Significant research effort has been targeted towards identification of the genes involved in permanent vision loss through photoreceptor and/or retinal pigment epithelium (RPE) cell dysfunction or cell death. So far, mutations in over 250 genes are known to be involved. Despite a good understanding of the genotype-phenotype relationship in AMD, this has not translated into predictive in vitro and in vivo models to understand disease mechanisms.

Partnership on iPSC-derived RPE cells
To drive the search for treatments for this devastating disease, it is vital to identify and develop new models which enable elucidation of retinal disease mechanisms and reliably predict the efficacy of therapeutic compounds. To address these challenges, Evotec has teamed up with the Centre for Regenerative Therapies (CRTD) in Dresden, Germany. The CRTD has a longstanding interest in degenerative processes and has contributed significantly to our understanding of retinal diseases in recent years. The joint TargetRD project is based on Evotec’s know-how in induced pluripotent stem cell (iPSC) technology, generating iPSC-derived RPE cells from AMD patients.

RPE cells are essential for visual function and a key component for light detection by photoreceptor neurons. They also are crucial for maintaining the blood-retina-barrier, for the transport of diverse biomolecules, ions, and fluids in and out of the retinal tissue, and recycling of the visual chromophore retinal molecule.

Even more important is their ability for phagocytosis: A single RPE cell is in contact with about 30 photoreceptor cells and responsible for the phagocytosis and removal of the distal portions of the photoreceptor outer segments that are phagocytozed in the course of a day. This is an important process with up to 10% eliminated daily, meaning the entire population of photoreceptor outer segments is turned over every 2 weeks. Maintaining, repairing, or replacing RPE cells therefore is crucial for the management of AMD, but also for other retinal degenerative diseases.

So far, research has been hampered by limited access to RPE cells. There are immortalized retinal cell lines available, but they lack the typical cell morphology or function of their in vivo counterparts, e.g. pigmentation, polarization, and expression of certain proteins. Likewise, artificial organoids, so-called 3D-cups, are not ideal since they are difficult to grow and differentiate and not suited for high-throughput profiling of compounds.

This situation is now improving as Evotec and CRTD succeeded in developing a protocol to efficiently and robustly produce high quality human iPSC-derived RPE cells at industrial scale from patient cells. This means that partners can not only investigate disease pathology directly in this highly relevant retinal cell type but for the first time also study disease phenotypes and mechanisms within the context of a patient’s genome. Moreover, the TargetRD platform enables the study of individual, overlapping functional and morphological changes from iPSC-RPE cells derived from patients with different genetic backgrounds, providing an opportunity to unravel complex AMD disease phenotypes.

Promising first results
Already, the partners were able to show the utility of the TargetRD platform by analyzing iPSC-RPE cells carrying a patient mutation resulting a lysosomal storage disorder. Amongst other symptoms, patients with this type of mutation develop severe retinal degeneration, leading to complete blindness early in life. Using the various phenotypic and functional assays established, it was possible to demonstrate impaired trans-epithelial resistance in patient cells. This indicates that in this case, patient RPE cells are unable to form the tight monolayer required for normal RPE cell function. Furthermore, patient RPE cells have, as expected, impaired lysosomal activity and are unable to phagocytose photo- receptor outer segments (POS) at the same rate as control RPE cells. Since all assays are able to support high-throughput screening, the partners can use the TargetRD platform to identify phenotypic and functional disease phenotypes from patient cells, enabling novel drug discovery approaches.

Furthermore, the project combines Evotec’s drug discovery expertise and the academic excellence of the CRTD to achieve significant progress towards developing therapies much needed by the patients.

Both partners believe this is a very promising approach that enables successful drug discovery programs for retinal degenerations coupled to a high likelihood of successful translation into the clinic.

The dispensation of compounds in aqueous or DMSO solutions is an integral part of Evotec’s sample management processes. The company has the capability for the automated array of large numbers of samples between a large variety of different containers suitable for both, distribution or storage. Its extensive liquid handling resources cover the widest range of possible volumes.

High-volume systems can dispense and transfer from 1 uL to multi-milliliters to and from vials and plates, while mid-volume systems can dispense from 1 uL to 2 mL using 96, 384 and 1536 plates or racks (SBS - Society for Biomolecular Screening – formats).

As Evotec is striving towards the goal of getting drugs to patients faster, it is constantly seeking to improve the speed of screening cycles and data quality. To enable a step change in drug discovery research and development and to obtain results faster and even more accurately with very small sample sizes, Evotec recently invested in the so-called Acoustic Tubes technology in partnership with Brooks, Beckman and Titian. Acoustic sampling has revolutionised liquid handling and Evotec is using two different systems for acoustically dispensing volumes over 1 uL, or between 2.5 nl and 1 µl.

The technology is using ultrasonic acoustic energy, where energy focused just below the meniscus of a liquid sample ejects small droplets of liquid, enabling very small volumes to be transferred from source wells and precisely positioned onto a destination surface suspended above the ejection point. As a result, scientists can do more with less, handling multiple workflows and generating faster results without the restrictions of traditional tip-based liquid handling.

Moreover, this acoustic technology is combined with newly designed caps, a new de-capping and recapping technology hence creating the ability to store and access compounds long-term directly from the same tubes.

To evaluate the effects of acoustic labware use for long and short-term storage as well as the impact of freeze/thaw cycles and acoustic dispensing on compound integrity, Evotec has completed rigorous testing of the technology before adopting it.

The technology has a number of benefits. Most importantly, it speeds up screening cycles and improves the capacity for cherry-picking plate creation. Besides minimising compound consumption, it also works without disposable tips. This reduces waste, operating costs and eliminates the risk of cross-contamination between compounds. Moreover, its precision and speed leads to unsurpassed accuracy and repeatability of results, while maintaining sample integrity and viability during the transfer. All in all, it saves time, cost and resources, provides for highest possible precision, and increases the flexibility in transferring liquids at different positions and volumes.

Adoption of acoustic sampling technologies increases the flexibility of the compound library at Evotec and its partners and speeds up the throughput for assay-ready plate preparation while reducing the consumption of compounds. Evotec is offering direct access to this technology to its customers on demand.

LEARN MORE

Monoclonal antibodies (mAbs) have been a huge success story in the biopharmaceutical industry. They have changed the landscape of biologicals and offered therapies for previously untreatable diseases. Although mAbs have been on the market since the mid 1980s, they have become increasingly sophisticated over the past decades. This also holds true for the technologies enabling the identification, design, preclinical and manufacturing processes of monoclonal antibodies.

While the traditional hybridoma technology is still broadly used for the creation of mAbs, novel technologies such as immune antibody libraries or fully humanoid antibody libraries have emerged in recent years. Moreover, researchers can today leverage a range of powerful approaches to optimize the mAb creation process, e.g., high-throughput screening, state-of-the-art sequencing, AI / machine learning or analytics. This, in turn, speeds up the development cycle, allows for precise selection of the desired mAb properties and ultimately reduces failures of monoclonal antibodies at advanced development stages.

But how to choose from the vast range of technologies for antibody development? This challenge was discussed at Evotec´s recent Innovation Week in a session titled "Translate your idea into a product: AI -driven antibody discovery at Evotec".

The session covered Evotec´s capabilities for supporting all activities across the R&D continuum, i.e.

• Discovery and optimization of novel antibodies for specific disease targets,
• Evaluation of lead antibodies for disease efficacy and safety and
• Process development and manufacturing.

"In many cases, it is not the question of using either one or another technology," says Barbara Bachler-Konetzki, Group Leader In Vitro Pharmacology at Evotec. "Depending on the desired properties of the monoclonal antibody, combining several technologies will do the trick. Therefore, it is important to have access to a comprehensive repertoire of leading-edge, synergistic technologies."

For state-of-the-art monoclonal antibody development, Evotec has established a unique one-stop-shop from target identification to IND. This includes a broad technology platform as well as unparalleled expertise in drug development and even manufacturing, including latest advances in artificial intelligence and machine learning such as generative adversarial networks (GAN) to create synthetic realistic outcomes by machine learning (J.HAL).

The resulting integrated biologics platform is called J.DESIGN and integrates molecular, process and manufacturing design.

The example of Evotec´s internal SARS-CoV-2 campaign shows how its proprietary J.HAL technology can be used to identify antibodies effectively blocking the SARS-CoV-2 infection pathway by binding to SARS spike protein, effectively neutralizing the infectivity across several SARS-CoV-2 strains. The subsequent in silico sequence analysis informs about mAb properties and engineering opportunities to reach the desired properties re immunogenicity, stability etc. Further steps, e.g., sequence and stability optimization, improve manufacturability and yields or pharmacokinetics (PK).

For mAb development, Evotec pursues a translational approach, i.e., leveraging its extensive expertise in various therapeutic areas to facilitate the setup of disease-relevant biological assays. Among others, this allows for PK/PD characterization of biologics early in the R&D process as well as the prediction of downstream in vivo efficacy and demonstration of target engagement. Moreover, Evotec’s pre-clinical department offers the full range of in vitro and in vivo GLP and non-GLP pre-clinical evaluation studies to assess the safety profile of the drug candidate. In addition, Evotec has established several sophisticated manufacturing facilities worldwide to reduce the risk of downstream attrition and delay.

This unparalleled infrastructure and translational expertise put Evotec in a unique position to conduct leading-edge, integrated monoclonal antibody development programs all the way from target identification to manufacturing.

FULL REPLAY

Chemoprophylaxis is a well-known strategy to prevent infectious diseases in populations at risk, e.g., in malaria, tuberculosis, or Neisseria meningitidis. Therefore, it is also being discussed for other diseases, such as Chikungunya virus (CHIKV) infection. This disease is spread by Aedes mosquitoes and leads to acute febrile illness, as well as to inflammation and disabling pain in the joints. While the acute fever often resolves within days, musculoskeletal, arthritis-like symptoms can persist for months and years, leading to a substantial burden of disease in tropical and subtropical areas.

There is a need not only for therapeutic, but also prophylactic treatments against Chikungunya disease – like those available for malaria. However, the design of clinical trials is complex as Chikungunya outbreaks are challenging for studying interventions: They are unpredictable in time and place, spreading rapidly, but with usually short life-spans and differential diagnosis is complex, so that there is a high risk that the outbreak is over before a trial can be conducted.

However, there is a potential solution, following evidence on the risk of secondary household infections in chikungunya outbreaks. Household transmission is well-known from respiratory viruses transmitted via aerosols. At first glance the transmission via a mosquito seems not comparable to the transmission mode of airborne viruses, but it is known that the main vector, Aedes mosquitos, has a very limited flying range, resulting in spatial microclustering of CHIKV outbreaks: household members and near neighbors have the highest risk of secondary infections. Therefore, enrolling such a high-risk population could allow for a more feasible, smaller, shorter and conclusive trial.

Evotec teamed up with scientists from Aarhus University (Denmark), the Institut Pasteur du Cambodge (Phnom Penh, Cambodia), the Fundação Oswaldo Cruz, (Salvador, Bahia, Brazil), and the Instituto Nacional de Infectologia Evandro Chagas-Fiocruz (Rio de Janeiro, Brazil) to develop an innovative chemoprophylaxis trial design for CHIKV.

As a first step, such a trial requires a surveillance study design to determine household secondary attack rate. For this study, index cases need to be identified by RT-PCR to confirm a CHIKV infection, followed by serosurveillance of household members. There is a caveat as household secondary attack rates may not be accurately predicted from one chikungunya outbreak to another. Therefore, estimates of household secondary attack rates from different countries and cultural environments are needed.

The objective of such a study will be first to establish a range of estimates around which the feasibility of prophylaxis trials can be evaluated and a sample size calculated. Subsequently, evidence-based prophylaxis trials can be conducted based on the estimated rate of secondary household infections.

LEARN MORE

While the public has taken note of RNA-based medicine only with the advent of mRNA-based Corona virus vaccines, biopharmaceutical research and development has been working on mRNA-based medicine for almost two decades. Evotec also expanded the druggable target space to RNA and in the last years added considerable know-how in RNA-based medicine.

RNA is used by cells in multiple ways: mRNA is conveying genetic information from DNA to the ribosomes which also are made from RNA (ribosomal RNA), where another RNA species (tRNA) is transporting amino acids to the ribosomal apparatus so that a protein can be synthesized. In addition to mRNA, there are also shorter RNA molecules being used in the cell for the regulation of genes and entire genetic cascades.

This provides for plenty of potential interventions: antisense (ASO) and short interfering RNA (siRNA) can up or down regulate an RNA target, e.g., to block the translation of an unwanted or diseased protein or to suppress or stimulate the expression of genes. RNA can be targeted with (complementary) RNA, but it is also possible to alter or block the translation, re-locate or initiate RNA, degradation, etc. by small molecules interfering with the three-dimensional structure of RNAs or protein-RNA-complexes.

During our recent Innovation Week, Evotec experts Steffen Grimm, Group Leader, Hit ID & Biophysics, and Hilary Brooks, Senior Research Scientist, In Vitro Pharmacology, hosted a session called "The early bird catches the helix: Expanding the druggable target space to RNA".

In the session, they discussed how to:

• Expand the potential for drugs targeting RNA to offer alternative solutions for diseases with otherwise undrugged targets
• Target RNA providing highly specific solutions for protein removal, alternative splicing or pathway regulation via noncoding RNA
• Use the small molecule RNA targeting platform to contribute to new opportunities for target identification and validation

RNA as Therapeutics
Using RNA as therapeutics is not trivial. Nucleic acids introduced from outside may trigger adverse reactions by the innate immune system. A lot of knowledge is necessary to ensure delivery, avoid degradation and inflammation and to fine-tune the stability and function of the molecules. RNA may also have off-target effects. To ensure efficacy and safety, monitoring these early on needs to be incorporated into the developmental workflow. High quality synthetic RNA is costly to make, therefore a scaleable process and the relevant analytics must be established early in the process to accompany both the discovery and development stages of research with quality test material; Eventually producing GMP grade RNA at a commercial scale (several hundred grams) for human administration.

Evotec already has integrated all capabilities under one roof, allowing for the complete preclinical data set, reduced transition times and efficient communication to the regulators. For antisense oligonucleotide therapy, efficient hit sequences that knock down target expression can be selected in a matter of weeks. Toxicity profiling is a priority to establishing final leads and, subsequently, project-specific dose, duration and delivery will be established using optimized backbone chemistry. Using its in-silico capabilities as well as iPSCs, animal models, transcriptomics, etc. Evotec is able to predict toxicity and efficacy, and de-risk unwanted immune stimulation as well as off-target effects. For manufacturing, Evotec is discovery-capable and already building medium-scale capacity (up to 50g) which will be ready by 2023.

For inhibiting the translational machinery, Evotec has established an RNA small molecule targeting platform and established in various case studies, molecules binding to RNA, and demonstrating a significant effect in vitro without affecting cell viability. Evotec’s capabilities also allow the creation of a representation of the 3-dimensional structure of the target complex and its interaction with the compounds.

Evotec’s experienced team of scientists with proven drug discovery and development expertise already have a track record of driving RNA targeting projects forward. Its integrated medicinal and computational chemistry capabilities, combined with bioinformatics, structural biology, pharmacology, and drug safety expertise allows for the identification and characterization of RNA target species and their modulation by different modalities. Partner projects can be driven all the way from target identification to IND and beyond. Evotec therefore is a low-risk outsourcing partner and a company continually investing in its platform to the benefit of the customer.

LEARN MORE

The development of novel drugs is requiring much more than preclinical and clinical studies. Once scientists have identified a biological structure they want to block or to engage with, they begin screening for compounds that are able to fulfill this task. However, before they can even start this screening process, which can involve tens of thousands of compounds, these molecules have to be synthesized or extracted, purified, collected, stored, catalogued and tracked so that they can be made available to be processed.  

Compound management involves receiving and registering compounds from any external sources, organizing their long-term storage in a safe and controlled environment. For the acquisition of new molecules, Evotec has established a platform collecting and evaluating catalogs of compound providers. Compounds not yet available in Evotec’s collection can be sourced via this platform and added to the collection or made available to the customer as required. These compounds must be stored in a dedicated space under the right conditions in terms of temperature, humidity and other factors which could influence their stability. Access is restricted to authorized personnel to avoid accidental or deliberate manipulation of the samples. This requires a lot of logistics, regular controls, and supervision. 

To ensure high quality, safe, and speedy delivery of compounds, Evotec has established six compound management teams located in Europe and the US. Its main sites in Toulouse (France), and in Branford, (CT, USA), handle most of the samples received by Evotec, which include small molecules as well as biologicals. Overall, Evotec’s six compound management sites can store over 13 million compound containers under optimum conditions, and are able to process and deliver over 53 millions samples per year. The Company’s high quality compound storage and fast turnaround time, are based on robust automated technology, a highly sophisticated laboratory information management system (LIMS), and secure data management. All sites operate under quality management and internal auditing principles. The main sites  have developed a Business Continuity plan to ensure complete protection for our customers. Evotec is using Titian's Mosaic software, the leading software solution for sample management, providing  for a full audit trail and complete sample life cycle tracking. Every single sample carries a unique identifier barcode and the LIMS system is able to track each operations across all sample management processes, ensuring refill of depleted substances are automatically enacted.  The system has in-built redundancy to protect the data against critical loss. 

Once ordered, delivery of individual compounds or compound collections to customers and partners are organised in a short time frame ensuring the right format, concentration, and volume of substances are prepared. Samples are being shipped all over the world. Evotec also takes care of choosing the right parcel services, as frozen or temperature-sensitive substances need to be shipped under specific controlled conditions to protect temperature shift. In addition,  Evotec’s dedicated logistic team has expertise in documentation requirements, tariff classification, and import/export customs requirements to ensure compounds shipped can travel through customs without delays linked to incomplete paperwork.  

Evotec is continuously investing in state-of-the-art technologies to keep its compound management tools up to date. As an example, it invested in acoustic sample management, where liquids are transferred via acoustic sound waves, creating a droplet at the liquid’s surface. This technology increases flexibility, throughput and reduces compound consumption. The Company has been building screening libraries since 2004 and is serving a broad variety of customers ranging from academia, non-profit organizations to medium-sized biotech and big pharma. Apart from ordering and receiving compounds, customers  are also able to enter collaborations in which Evotec stores and manages their proprietary compound libraries and collections. This enables large companies as well as fast-growing SMEs to save space and costs while gaining more flexibility.  

As a result, customers profit from Evotec’s expertise, state of the art investment with large storage capacity and capabilities, its experienced staff and the inbuilt security safeguarding quality and availability of valuable compounds. Thereby, Evotec’s compound libraries and management tools significantly contribute to the success of drug discovery research projects as well as to medicinal product development. 

LEARN MORE

Evotec strives to provide partners and customers with less expensive, faster, and more flexible approaches for discovering, developing and manufacturing biotherapeutics.

With the acquisition of Just Biotherapeutics in 2019, Evotec made a major push into biologics, and has successfully built a fully-integrated platform to drive monoclonal antibody (“mAb”) programmes from concept through to commercialisation. The company generates antibody lead candidates by providing access to in vivo and in vitro sources of antibodies, combined with state-of-the-art technologies to ensure success for a broad range of targets and disease states. In addition, if needed, selected lead candidates can be further optimised using powerful computational platforms such as Evotec’s proprietary Abacus™ in silico tool suite to enhance the productivity, but also ease of manufacturing and formulation stability.

For in vivo immunisation, Evotec is offering the ATX-Gx^TM mouse platform developed by its collaboration partner Alloy Therapeutics. This platform is a suite of immunocompetent transgenic mice for best-in-class in vivo discovery of fully human monoclonal antibodies. These mice are engineered to drive the greatest potential diversity and broadest epitopic coverage of unique human antibodies binding to a specific target of interest.

ATX-Gx^TM mice provide:

• a fully human heavy chain repertoire,
• human kappa and human lambda chain repertoire,
• haplotype diversity, and
• limited immunodominance.

Taken together, these immunocompetent transgenic mice represent the complete functional human antibody repertoire, and their fully functional, robust immune response is equivalent to the response of wild type mice. They are also available on multiple genetic backgrounds such as BALB/c or BL/6.

ATX-Gx^TM has been extensively validated and continuously expanded for more than 12 years and is currently used by over 75 antibody discovery groups in large biopharma and small-to-midsize biotech companies as well as academic research labs.

Following immunisation, Evotec is combining traditional hybridoma technology with automated devices for high throughput clone selection, screening, recombinant expression and purification to shorten and simplify the process of hybridoma establishment. Screening is performed with high-throughput technologies, such as the iQue® Advanced Flow Cytometry Platform. Next-generation sequencing is used to obtain VH/VL information, followed by an in silico analysis using Just – Evotec Biologics Abacus® software tool. Once VH/VL information is available, several hundred mAbs can be produced in parallel in small scale. This high quality material can then be used for further downstream characterisation, such as functional activities that allow the selection of potential lead candidates.

This streamlined workflow to generate high-quality antibody panels for functional assessment adds to Evotec’s comprehensive suite of large molecule discovery tools, disease biology, state-of-the-art cell line development, machine learning design tools, manufacturing, preclinical IND-enabling studies, as well as FIH clinical support, and commercialisation.

Evotec’s AI-driven antibody discovery can be accessed as stand-alone services or through Evotec’s seamlessly integrated antibody drug discovery platform

LEARN MORE

The Covid-19 pandemic is a stark reminder that infectious diseases and their sometimes devastating effects will always have to be reckoned with. However, beyond Covid-19 a second global health crisis is emerging, and it is imperative that we act now to prepare for the increasing development of antimicrobial resistance (AMR) towards our currently existing arsenal of antibiotics.

The key to de-risking and expediting the development and approval of new antimicrobials is a detailed understanding of the relationship between the fate of the antimicrobial compound in the body: pharmacokinetics (PK), and the impact of exposure to the compound on the target microbe: pharmacodynamics (PD). This understanding is essential for development of optimal human dosing regimens, maximising efficacy and minimising the emergence of resistance. Only with this understanding will we mitigate the risk of clinical trial failure, and ultimately extending the clinical utility of a new antimicrobial in the face of increasing AMR.

Among the several non-clinical in vivo and in vitro models of infection which provide complementary information to understand this PK/PD relationship, the in vitro Hollow Fibre Infection Model (HFIM) is the most versatile. Evotec’s comprehensive HFIM capabilities combined with its drug development expertise and unique EvostrAIn™ collection of microbial pathogens provide a versatile in vitro PK/PD platform to de-risk and accelerate the development of antibacterial, antifungal and antiviral compounds.

The Hollow Fibre Infection System consists of two principle compartments:

1. a central reservoir and associated tubing, which constitutes a circulating system, and
2. a hollow fibre cartridge consisting of thousands of hollow permeable capillaries, or fibres, housing the bacteria, fungi or virus of interest

The HFIM is a dynamic model that is able to simulate almost any given concentration-time profile for an antimicrobial compound or combination of compounds, without the constraints of animal PK. The containment of the bacterial in the peripheral compartment of the hollow fibre cartridge means the system is able to simulate PK profiles with no bacterial cell washout. It is also suitable for simulated dose range fractionation studies and can determine resistance prevention exposure for any simulated PK profile. Combining the ability to run long-duration experiments with multiple drug infusion profiles means that the HFIM is especially well suited to the development of antimicrobial combination therapy against slowly replicating bacteria such as Mycobacterium tuberculosis, a view which is supported by the European Medicines Agency (EMA).

When should the Hollow Fibre Infection Model be used in the drug discovery process?

• Early screening of compounds can de-risk future HFIM experiments.
• Studies at the pre-clinical stage (pre-IND) will help to improve understanding of the PK/PD relationship. This is particularly important if an animal model is not available, or if the animal can’t tolerate the inoculum levels that need to be tested for example in resistance studies.
• During clinical development the HFIM can help to inform trial design. Hollow fibre studies can be performed based on human exposure data to address discrepancies between clinical and pre-clinical findings.
• Even after drug approval the HFIM can be used to expand the label or optimise the dosing regimens.

Evotec has established HFIM capabilities in state-of-the-art containment level 2 facilities, with a growing team of dedicated Hollow Fibre specialists. Providing full microbiological support to projects and aided by specialist bioanalytical and mathematical modelling and simulation teams, Evotec can offer a bespoke in vitro PK/PD service tailored to advance individual antimicrobial development programmes.

Learn more about Evotec's Hollow Fibre Infection Model by downloading our white paper

DOWNLOAD NOW

The Covid-19 worldwide pandemic unveiled extraordinary resources within the scientific and pharmaceutical community delivering vaccines and therapeutic drug candidates within timelines never seen before. While the pandemic hit suddenly and the number of cases increased exponentially, the AMR crisis remained silent. Nevertheless, it is progressing and the emergency and spread of multidrug resistance is putting our existing antibiotic arsenal under increasing threat.

Is AMR our next pandemic?

Possibly, if we continue to consider antibiotics as fire-extinguishers: and only create novel therapeutics once we are face to face with the fire.

Innovation and collaboration are key in the preparedness, together with expertise and operational excellence. Evotec, with more than 200 scientists dedicated to antibacterial drug R&D, is at the forefront of AMR research and innovation. With infectious disease platforms spanning from in vitro biology to in vivo pharmacology and medicinal chemistry and benefiting from HTS platforms, ADME and toxicology expertise.

Innovative thinking and creativity to discover novel antibacterial compounds begins with designing a strategy to discover novel active compounds. By changing the paradigm in phenotypic screening and developing the Vivo Mimetic Media (VMM) concept for discovering novel Gram-negative antibacterials, Evotec has validated five alternative bacterial culture media that better mimic the conditions bacteria are facing during infections. These VMM conditions are affecting the physiology of growing bacteria by altering permeability (porins, efflux pumps and outer membrane) but mainly unveiling new targets and mechanisms of action (MoAs).

The cornerstone of innovation in antibacterial drug discovery is the application of machine learning to optimise chemical matter. By combining biological data, medicinal chemistry expertise and a deep learning approach, Evotec has enabled the prediction of antibacterial activities against 15 bacterial strains and provided de novo design of compounds, 76% of which were accurately predicted for their activity.

Validation of active compounds in appropriate pharmacodynamic models is crucial, while innovative approaches are needed for tailored design and improved readouts. By using luminescent or fluorescent bacteria, infection models with higher complexity have been developed to evaluate the spread of infection in real time and most importantly, measure the distribution of the active compound fluorescently labelled to the site of infection.

Innovation through collaboration and partnership is at the forefront of Evotec’s unique business model. Through a highly strategic partnership between Evotec, Resolute Therapeutics and CARB-X, a new antibacterial drug class termed TriBe is being advanced in the preclinical pipeline, with the objective to bring a candidate for the treatment of cUTI, cIAI and lung infections to clinical phase I. The TriBE series have unique properties with nanomolar bacterial topoisomerase inhibitors binding via the GyrB/ParE subunits, very broad bacterial spectrum, low potential to select for resistance, favourable PK and in vivo efficacy in multiple models of infection have been demonstrated.

While a consensus exists on the real and clear need for new antibacterials and approaches, the challenges are numerous and complex. Innovative thinking, creativity and novel approaches (target and technology) underpin anti-infective drug discovery at Evotec, enabling the redesigning of antibacterial discovery infrastructure to ensuring an integrated approach and foster collaboration, partnership, training and cross-interaction.

WATCH OUR WEBCAST TO LEARN MORE