Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery

Mateus, André; Gordon, Laurie J.; Wayne, Gareth; Almqvist, Helena; Axelsson, Hanna; Seashore‐Ludlow, Brinton; Treyer, Andrea; Matsson, Pär; Lundbäck, Thomas; West, Andrew; Hann, Michael M.; Artursson, Per

doi:10.1073/pnas.1701848114

Cited by 75 publications

(92 citation statements)

References 70 publications

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“… 26 Finally, the physicochemical properties of PDP 3 could be limiting its intracellular free concentration. 37 Given these variables, designing a PDP against a protein target that has not previously been validated as being susceptible to E3 ligase-directed degradation is challenging, as optimization cycles can appear lengthy and the difficult and low yielding synthesis discourages the generation of multiple analogues. It was also unclear how strict and narrow the requirements for optimal PDP design would be.…”

Section: Results and Discussionmentioning

confidence: 99%

Demonstrating In-Cell Target Engagement Using a Pirin Protein Degradation Probe (CCT367766)

et al. 2018

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Demonstrating intracellular protein target engagement is an essential step in the development and progression of new chemical probes and potential small molecule therapeutics. However, this can be particularly challenging for poorly studied and noncatalytic proteins, as robust proximal biomarkers are rarely known. To confirm that our recently discovered chemical probe 1 (CCT251236) binds the putative transcription factor regulator pirin in living cells, we developed a heterobifunctional protein degradation probe. Focusing on linker design and physicochemical properties, we generated a highly active probe 16 (CCT367766) in only three iterations, validating our efficient strategy for degradation probe design against nonvalidated protein targets.

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Section: Results and Discussionmentioning

confidence: 99%

Demonstrating In-Cell Target Engagement Using a Pirin Protein Degradation Probe (CCT367766)

et al. 2018

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“…Directing the drug to localize to the relevant organelle within the pathogen by altering its chemical structure and properties has the potential to increase its pharmacological activity. Accordingly, improving the ability of a small molecule to localize to the specific organelle or domain that harbors its target can significantly improve its efficacy due to an increase in the local concentration of the inhibitor …”

Section: Figurementioning

confidence: 99%

Localizing Antifungal Drugs to the Correct Organelle Can Markedly Enhance their Efficacy

et al. 2018

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A critical aspect of drug design is optimal target inhibition by specifically delivering the drug molecule not only to the target tissue or cell but also to its therapeutically active site within the cell. This study demonstrates, as a proof of principle, that drug efficacy can be increased considerably by a structural modification that targets it to the relevant organelle. Specifically, by varying the fluorescent dye segment an antifungal azole was directed from the fungal cell mitochondria to the endoplasmic reticulum (ER), the organelle that harbors the drug target. The ER‐localized azole displayed up to two orders of magnitude improved antifungal activity and also dramatically reduced the growth of drug‐tolerant fungal subpopulations in a panel of Candida species, which are the most prevalent causes of serious human fungal infections. The principle underlying the “target organelle localization” approach provides a new paradigm to improve drug potency and replenish the limited pipeline of antifungal drugs.

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“…For in vitro cellbased assays, such as metabolic stability, induction, inhibition, and pharmacological assays, f u measurements of hepatocytes or other cell types [ f u of cells ( f u,cell )] allows for the determination of intracellular free drug concentration (Mateus et al, 2013;Riccardi et al, 2016Riccardi et al, , 2017. Intracellular free drug concentration, rather than nominal concentration, is most relevant for compounds with intracellular accumulation or exclusion to develop in vitro-in vivo correlations for human translation and to understand the in vitro absorption, distribution, metabolism, excretion, and toxicity and pharmacology endpoints (Riccardi et al, 2016(Riccardi et al, , 2017Mateus et al, 2017;Riede et al, 2017;Sun et al, 2017). Using intracellular free drug concentration, the unbound partition coefficient (K puu ) can be determined and used to derive intrinsic activity for in vitro cell-based assays (e.g., CL int = CL int 9/K puu , EC 50 = EC 50 9 Â K puu , IC 50 = IC 50 9 Â K puu , where CL int is intrinsic clearance, CL int 9 is apparent intrinsic clearance, EC 50 9 is apparent EC 50 , and IC 50 9 is apparent IC 50 ).…”

Section: Introductionmentioning

confidence: 99%

Comparison of Species and Cell-Type Differences in Fraction Unbound of Liver Tissues, Hepatocytes, and Cell Lines

et al. 2018

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Fraction unbound () of liver tissue, hepatocytes, and other cell types is an essential parameter used to estimate unbound liver drug concentration and intracellular free drug concentration. and are frequently measured in multiple species and cell types in drug discovery and development for various applications. A comparison study of 12 matrices for and of hepatocytes in five different species (mouse, rat, dog, monkey, and human), as well as of Huh7 and human embryonic kidney 293 cell lines, was conducted for 22 structurally diverse compounds with the equilibrium dialysis method. Using an average bioequivalence approach, our results show that the average difference in binding to liver tissue, hepatocytes, or different cell types was within 2-fold of that of the rat Therefore, we recommend using rat as a surrogate for liver binding in other species and cell types in drug discovery. This strategy offers the potential to simplify binding studies and reduce cost, thereby enabling a more effective and practical determination of for liver tissues, hepatocytes, and other cell types. In addition, under hepatocyte stability incubation conditions should not be confused with , as one is a diluted and the other is an undiluted Cell density also plays a critical role in the accurate measurement of.

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Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery

Cited by 75 publications

References 70 publications

Demonstrating In-Cell Target Engagement Using a Pirin Protein Degradation Probe (CCT367766)

Demonstrating In-Cell Target Engagement Using a Pirin Protein Degradation Probe (CCT367766)

Localizing Antifungal Drugs to the Correct Organelle Can Markedly Enhance their Efficacy

Comparison of Species and Cell-Type Differences in Fraction Unbound of Liver Tissues, Hepatocytes, and Cell Lines

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