Science Pool

As our understanding of conformational chemistry advances, we can explore a variety of strategies beyond cyclization to restrict the number of low-energy conformations accessible to ligands. These alternative approaches offer the potential to optimize binding affinity and enhance the overall pharmacological profile of ligands, facilitating more efficient drug design and discovery.

In this comprehensive review we highlighted the essential role of conformational drivers in drug design, exploring how molecular conformation influences key drug properties with real-world examples.

Targeting RNA represents a paradigm shift for drug discovery. The ability to seek out and destroy, or modify, a faulty RNA template, before the toxic protein has even been made, has only recently begun to be harnessed for the benefit of patients. 
At the time of writing, 21 Oligonucleotide drugs have been approved for human use, with an exponential increase in clinical trials and development projects involving this new modality. 
There exist several different mechanisms of action for Oligonucleotide drugs, all of which are transient and reversible and do not lead to alteration of patient DNA, unlike Gene therapy. 

Antisense Oligonucleotides harness endogenous systems already existing within a cell to achieve their purpose, with the only limitation being accessibility of the target tissue.
Once bound with great specificity to its RNA target, a short synthetic Oligonucleotide can trigger degradation, upregulation of the translated protein, or alteration of a splicing event leading to a correctly folded protein. Longer Oligonucleotides can fold into 3 dimensional shapes called Aptamers with similar target affinities and applications as antibodies, and shorter Oligonucleotides can act as MicroRNA   mimetics or antagonists to alter multiple targets or pathways concurrently with subtle but broader effect. 

The precision of an Oligonucleotide and its ability to correct a faulty RNA produced by an error in the genetic code, lends itself to applications in the fields of rare disease therapeutics and toxic gain of function mutations. The field of Oligonucleotide therapeutics is developing to address this as a whole and to pioneer a new preclinical and regulatory path that could be adapted for these unique disease biologies to make this type of therapeutic innovation more accessible. 

Evotec is a leader in integrated end-to-end Research and Development and has built substantial drug discovery expertise and technical capabilities that can drive new innovative and diverse modalities into the clinic. In addition, Evotec has developed a deep internal knowledge base in key therapeutic areas including neuroscience, pain, immunology, respiratory, women’s health, aging, fibrosis, inflammation, oncology, metabolic and infectious diseases. Leveraging these skills and expertise, Evotec successfully delivers on superior science-driven discovery and development alliances with pharmaceutical and biotechnology companies.

The global interest in this new modality area has led to high demand in Oligonucleotide synthesis and related chemistry applications, from modified Oligonucleotides to conjugates and complex formulations. 
Evotec offers Oligonucleotide research and development capabilities as well as ligand and linker chemistry expertise to support projects from discovery through to development.

In particular, since 2015 the discovery chemistry group is equipped with MerMade 48X synthesizers (Biosearch Technologies, see Fig. 1) with associated purification, post processing and QC platforms (at the Evotec sites of Toulouse and Verona, see Fig 2), while the development team, since 2022, is equipped with a Cytiva ÄKTA oligosyntTM synthesizer associated to the ÄKTA pureTM 150 and ÄKTA flux 6 purification platforms and with a Virtis Lyophilizer at Evotec’s state-of-the-art Verona site (see Fig. 3 to Fig. 6).
	Figure 1: Biosearch MerMade 48X	Figure 2: Agilent Preparative HPLC for purification
	Figure 3: Cytiva ÄKTA oligosyntTM	Figure 4: Cytiva ÄKTA pureTM 150
	Figure 5: Cytiva ÄKTA flux 6TM	Figure 6: SP Virtis Advantage Pro Lyophilizer
	Figure 7: UPLC with a Thermo ScientificTM Orbitrap ExplorisTM 120 mass spectrometer for oligonucleotides characterization

This fully integrated suite of capabilities allows for the synthesis, purification, isolation, and quality control of complex modified Oligonucleotides (ASOs, siRNAs, etc) on a scale from milligrams up to 25 g (up to 12 millimoles). The objective is to support Oligonucleotide drug discovery and development projects from the earliest phases of discovery, such as the generation of screening libraries, up to the selection of a preclinical development candidate followed by manufacture and release of material to support initial preclinical development studies. 

All these activities are supported by an experienced Oligonucleotide chemistry team operating across two sites and at different scales, to ensure flexible support for projects with highly efficient information and process transfer.
Evotec capabilities also include expert analysts for Analytical Development and QC, capable of developing and validating the analytical procedures needed for a full characterization and routine testing of Oligonucleotide drug substances up to and including IND enabling studies. In addition, Evotec’s support can encompass the release of preclinical batches according to regulatory requirements, including stability and formulation studies.

The journey to commercialization can be challenging. Scaling up production while maintaining process consistency, product quality, and regulatory compliance, requires expert process development capabilities, and the adoption of innovative science and risk management methodologies. A common pitfall for the Sponsor of an innovative therapy is to under-estimate the complexity and intricacy of this enterprise, which involves the coordinated optimization of strategies for process control, risk management, data management, and supply chain management.

With ever-evolving regulatory requirements and the increasing urge to shorten drug development timelines, getting your drug to market can seem like a daunting undertaking. That’s why taking some of the pressure off your organization by outsourcing your drug development and manufacturing activities to an expert partner can be the smartest decision. This will ensure your drug is commercialized in the fastest and most cost-efficient way possible, utilizing expertise, facilities, equipment, and processes to anticipate and overcome any challenges thrown at your program with ease.

Evotec offers an integrated end-to-end solution for innovative drug R&D, with the capabilities to support all phases of your drug development program. Your projects are in safe hands with our team of expert scientists who are pioneers in QbD, process design, scale-up, and validation, operating to full cGMP within FDA, MHRA, AIFA and BfArM approved facilities.

Our experts are just a click away! 

Reach out to us today

Don’t miss our educational webinar series on “Oligonucleotides Therapeutics: Discovery to Development”

Register Here

As the world increasingly turns its focus toward sustainability, the pharmaceutical industry is not exempt from this shift. One company leading the charge is Evotec, a global drug discovery and development biotech who offer CRO and CDMO services. Evotec’s commitment to green chemistry is setting new standards for sustainability in medicinal chemistry, particularly in how drug discovery processes are designed and executed. This blog post will delve into Evotec’s green chemistry initiatives and how they are revolutionizing the pharmaceutical industry.

Evotec’s Commitment to Green Chemistry

Evotec’s green chemistry philosophy centres around minimizing the environmental impact of their chemical processes while maintaining excellence in drug discovery. This commitment is evident in their comprehensive strategy that encompasses various innovative approaches and technologies designed to make their operations more sustainable.

Key Initiatives in Green Chemistry

1. Safer Solvents and Reduced Plastic Waste
   • Evotec prioritizes the use of safer solvents, particularly those sourced from renewable bio-based origins, to reduce the reliance on chlorinated solvents known for their environmental and health hazards.
   • They actively work on reducing plastic waste by encouraging the reuse and recycling of plastic consumables in their laboratories.
2. Energy Efficiency
   • Energy conservation is a significant aspect of Evotec’s sustainability efforts. Simple yet effective practices like shutting laboratory fume cupboards when not in use help in reducing unnecessary energy consumption.
3. Raising Awareness and Continuous Improvement
   • Evotec fosters a culture of sustainability by raising awareness among their scientists about green chemistry alternatives through communication and poster sessions.
   • Continuous improvement of chemical processes with a sustainable vision is a cornerstone of their green chemistry initiatives.

Advanced Green Chemistry Techniques

Evotec employs several advanced green chemistry techniques that underscore their innovative approach to sustainable drug discovery:

1. Micellar Chemistry
   • Utilizing micellar properties for chemical reactions offers a new paradigm in sustainable chemistry, reducing the need for harmful organic solvents.
2. Mechanochemistry
   • This technique uses mechanical forces to drive chemical reactions, which can be more sustainable than traditional methods reliant on solvents and reagents.
3. Flow Chemistry
   • Flow chemistry enhances productivity and safety, offering better control over reaction conditions and significantly reducing organic waste.
4. Biocatalysis
   • By employing enzymatic processes, Evotec can replace traditional organic synthesis routes, reducing the use of toxic reagents and conditions, whilst often also increasing yields.

Practical Applications and Case Studies

Evotec has documented numerous case studies showcasing the practical application of green chemistry in their labs. These include:

1. Solvent Replacement
   • Evotec has successfully replaced conventional hazardous solvents like DMF, DCM, and THF with greener alternatives such as Me-THF, CPME and dimethyl isosorbide (DMI), achieving comparable or superior yields in various reactions while reducing toxicity and environmental impact.
2. Catalyst Optimization
   • By optimizing catalyst loading in reactions, such as pallado-catalyzed cross-couplings, the company has significantly reduced the amount of precious metals required, decreasing both environmental impact and costs.
3. Sustainable Work-Up Methods
   • The adoption of FastWoRX™, a technique that significantly reduces the amount of solvent used in the work-up phase, demonstrates Evotec’s commitment to minimizing waste.
4. Better purification processes
   • Rethinking our working habits in a smarter and sustainable way. Avoid systematic normal phase flash column chromatography (or reuse, if necessary). Favour reverse phase and crystallization/recrystallization.

The Future of Green Chemistry at Evotec

Evotec’s approach to green chemistry is not static; it is an evolving process that aims to continually integrate new technologies and methodologies. The company's dedication to sustainability is reflected in their global initiatives and partnerships, fostering a greener pharmaceutical industry.

In conclusion, Evotec is at the forefront of green chemistry in drug discovery. Their innovative approaches and commitment to sustainability set a benchmark for the industry, proving that excellence in drug discovery and environmental stewardship can go hand in hand. As Evotec continues to develop and implement green chemistry practices, they pave the way for a more sustainable future in medicinal chemistry.

LEARN MORE

Inhibition of NF-κB inducing kinase (NIK) has been pursued as a promising therapeutic target for autoimmune disorders due to its highly regulated role in key steps of the NF-κB signaling pathway. Previously reported NIK inhibitors from our group were shown to be potent, selective, and efficacious, but had higher human dose projections than desirable for immunology indications. Herein we report the clearance-driven optimization of a NIK inhibitor guided by metabolite identification studies and structure-based drug design. This led to the identification of an azabicyclo[3.1.0] hexanone motif that attenuated in vitro and in vivo clearance while maintaining NIK potency and increasing selectivity over other kinases, resulting in a greater than ten-fold reduction in predicted human dose.

READ NOW

At Evotec we are committed to the development and usage of new technologies. With the objective to stay at the cutting edge of science and propose innovative solutions, several working groups have been created and are actively involved in various fields: photochemistry, electrochemistry, flow chemistry, biocatalysis... As part of this strategy, the working group “green chemistry” aims to design chemical products and processes that reduce or eliminate the use or generation of hazardous substances. We are always looking for safer, greener and cleaner methodologies to reduce the environmental impact of our activities and adopt the green chemistry principles¹. Green chemistry applies across the life cycle of a chemical product, including its design, manufacture, use, and disposal. Moreover, we are also engaged in energy saving to decrease the global carbon footprint of the company. Sustainability and green chemistry are implemented while maintaining our level of excellence in drug discovery. To reach our objectives, we have identified four areas of improvements:

This poster is focused on two areas in continuous improvement at Evotec: solvent alternatives and energy saving. Some examples of reactions carried out in renewable solvents such as MeTHF² and DMI³ are presented. Alternatives to DCM (potential ozone depletory and suspected carcinogenic solvent) usage for work-up and purification are also shown⁴.

The poster was presented by Kim Spielmann at the Journées de Chimie Organique held on 2-4 November 2022 at the École Polytechnique, Palaiseau, France.

¹Anastas, P. T.; Warner, J. C. Green Chemistry: Theory and Practice, Oxford University Press: New York, 1998, 30, By permission of Oxford University Press
² a) Coby J. Clarke, Wei-Chien Tu, Oliver Levers, Andreas Bröhl, and Jason P. Hallett Chemical Reviews 2018, 118, 747 b) Pace, V., Hoyos, P., Castoldi, L., Domínguez de María, P. and Alcántara, A.R. ChemSusChem 2012, 5, 1369 c) Andrew Jordan, Callum G. J. Hall, Lee R. Thorp, and Helen F. Sneddon Chemical Reviews 2022, 122, 6749
³ a) Aricò, F.; Tundo, P. Beilstein J. Org. Chem. 2016, 12, 2256 b) F. Aricò, A. S. Aldoshin, P. Tundo, ChemSusChem 2017, 10, 53 c) Russo F., Galiano F., Pedace F., Aricò F., and Figoli A. CS Sustainable Chem. Eng. 2020, 8, 1, 659
⁴ a) Peterson E.A., Dillon B., Raheem I., Richardson P., Richter D., Schmidte R. and Sneddon H.F. Green Chem., 2014,16, 4060 b) Taygerly J.P., Miller L.M., Yeec A. and Peterson E.A. Green Chem., 2012,14, 3020 c) MacMillan D.S., Murray J., Sneddon H.F., Jamiesona C. and Watson A.J.B. Green Chem., 2012,14, 3016

DOWNLOAD

Targeting RNA represents a paradigm shift for drug discovery. The ability to seek out and destroy, or change, a faulty RNA template, before the toxic protein has even been made, has only recently begun to be harnessed for the benefit of patients.
As of this blog, only 16 oligonucleotide drugs have been marketed, with an exponential increase in clinical trials and development exploding in this area.
There exist different mechanisms of action for an oligonucleotide drug, all of which are transient and reversible effects and do not include alteration of the DNA, unlike Gene therapy.

Antisense Oligonucleotides harness endogenous systems already existing within a cell to achieve their purpose, with the only limitation being accessibility of the target tissue. Once bound with great specificity to its RNA target, a short synthetic oligonucleotide can cause degradation, upregulation of the translated protein, or alteration of a splicing event leading to correctly folded protein. Longer Oligonucleotides can fold into 3 dimensional shapes called Aptamers with similar target affinities and applications as antibodies, and shorter oligonucleotides can act as miR mimetics or antagonists to alter multiple targets or pathways at the same time with subtle but broader effect.

The precision accuracy of an oligonucleotide and its ability to correct a faulty RNA produced by an error in the genetic code, lends itself to the rare disease therapeutic area and toxic gain of function mutations. The field of oligonucleotide therapeutics is moving to address this as a whole and to innovate a new preclinical and regulatory path that could be adapted for these more unique diseases to make this type of therapy more accessible.

Evotec is a leader in integrated Research and Development (EVOiR&D) and has built substantial drug discovery expertise and technical capabilities that can drive new innovative, diverse modalities into the clinic. In addition, Evotec has built a deep internal knowledge base in key therapeutic areas including neuroscience, pain, immunology, respiratory, women’s health, aging, fibrosis, inflammation, oncology, metabolic and infectious diseases. Leveraging these skills and expertise, Evotec successfully delivers on superior science-driven discovery alliances with pharmaceutical and biotechnology companies.

The global interest in this new modality area has led to high demand in oligonucleotide synthesis and the chemistry surrounding it, such as covalent linkages and complex formulations. Evotec has oligonucleotide manufacturing capabilities as well as ligand and linker chemistry expertise to support discovery projects and is now expanding its capacity to support development stage oligonucleotide projects.

We are extremely proud to share that we have installed the first Cytiva AKTA oligosyntTM in Europe at our Evotec site in Verona.
This new state of the art equipment will allow for the synthesis of complex modified oligonucleotides (ASOs, siRNAs etc) on a scale from 0.5 to 50 g (up to 12 millimoles) to support the initial preclinical development studies.

This is a key milestone for Evotec, and, together with the brand-new AKTA flux 6 and AKTA Pure 150, this new oligonucleotide synthetiser will complete the fully integrated oligo suite Evotec Campus Levi-Montalcini in Verona. Currently both the Verona and Toulouse Evotec sites are equipped to support Drug Discovery programmes in the RNA therapeutics field with the synthesis of oligonucleotides on a research scale and now, Evotec, at the Verona site, has the capability to also support the preclinical and clinical development studies, i.e. analytical and bioanalytical activities.

We look forward to discussing with you – our partners from new or existing collaborations- how we can best help your oligo project succeed.

Get in Touch with Our Experts

The NADase SARM1 (sterile alpha and TIR motif containing 1) is a key executioner of axon degeneration and there is great interest in developing SARM1 enzyme inhibitors as candidate therapies for multiple neurodegenerative diseases.

Through a combined approach of X-ray crystallography and cryo-EM we uncovered the molecular mechanism of SARM1 inhibition by a compound (DSRM-3716), demonstrating that it undergoes a base-exchange reaction with the nicotinamide moiety of NAD+, and that the transferase product is the bona fide inhibitor. Further more we reveal the activated state of SARM1 for the first time.

LEARN MORE

Huntington´s Disease (HD) is a hereditary neurodegenerative disorder that is caused by a mutation in the huntingtin gene (mHTT) leading to deposition of pathologic protein aggregates in the brain.

This paper focuses on:

• Visualization of mHTT aggregate expression by positron emission tomography (PET) in rodent HD model
• Utility of the previously described novel PET imaging ligand CHDI-180 for detection of mHTT in rodent and post-mortem human HD brain
• Ability of CHDI-180 to serve as functional response indicator for mHTT lowering therapies

GET THE PAPER

Learn more about fighting tuberculosis by targeting the essential enzyme PptT.

Together with collaborators at Weill Cornell Medical College, Texas Agricultural and Mechanical University (TAMU), and  University of North Carolina (UNC), we identified the amidinourea compound AU8918, through a phenotypic screen, as an interesting anti-TB compound. We went on to identify its target within Mycobacterium tuberculosis, as phosphopantetheinyl transferase (PptT) - an essential enzyme involved in synthesis of cellular lipids and virulence factors.

This paper describes:

• Next steps taken in further pursuit of this novel anti-TB series
• An expanded collaboration within the TB Drug Accelerator consortium
• Latest structure guided SAR exploration around AU8918 for more potent and safest Lead-like analogs

GET THE PAPER

SARM1 is a NADase whose action triggers the destruction of axons and development of novel SARM1 inhibitors which enables the prevention or delay of neurodegenerative disorders. This paper identifies the binding site for the SARM1 agonist NMN and reveals the structure of full-length SARM1 as elucidated by cryo-electron microscopy (cryo-EM). The structure of apo SARM1 was revealed as an octameric ring, held in an autoinhibitory state by the separation of the active TIR domain at the rim of the ring. The elucidation of autoinhibition release and the identification of the NMN binding site within the autoinhibitory ARM domain opens a path to inhibition of SARM1 via stabilisation of its inactive form. These studies were possible through the close co-ordination of the new cryo-EM team at Evotec, Abingdon with Disarm and academic collaborators in Australia and the USA.

Proposed SARM1 activation mechanism:

LEARN MORE