Halogenation at the Phenylalanine Residue of Monomethyl Auristatin F Leads to a Favorable cis/trans Equilibrium and Retained Cytotoxicity.

Abstract

Halogenation can be utilized for the purposes of labeling and molecular imaging, providing a means to, e.g., follow drug distribution in an organism through positron emission tomography (PET) or study the molecular recognition events unfolding by nuclear magnetic resonance (NMR) spectroscopy. For cancer therapeutics, where often highly toxic substances are employed, it is of importance to be able to track the distribution of the drugs and their metabolites in order to ensure minimal side effects. Labeling should ideally have a negligible disruptive effect on the efficacy of a given drug. Using a combination of NMR spectroscopy and cytotoxicity assays, we identify a site susceptible to halogenation in monomethyl auristatin F (MMAF), a widely used cytotoxic agent in the antibody-drug conjugate (ADC) family of cancer drugs, and study the effects of fluorination and chlorination on the physiological solution structure of the auristatins and their cytotoxicity. We find that the cytotoxicity of the parent drug is retained, while the conformational equilibrium is shifted significantly toward the biologically active trans isomer, simultaneously decreasing the concentration of the inactive and potentially disruptive cis isomer by up to 50%. Our results may serve as a base for the future assembly of a multifunctional toolkit for the assessment of linker technologies and exploring bystander effects from the warhead perspective in auristatin-derived ADCs.

• Authors: Iris K Sokka, Surachet Imlimthan, Mirkka Sarparanta, Hannu Maaheimo, Mikael P Johansson, Filip S Ekholm
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 18 Issue 8 Pg. 3125-3131 (08 02 2021) ISSN: 1543-8392 [Electronic] United States
• PMID: 34296616 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Aminobenzoates
• Antineoplastic Agents
• Cytotoxins
• Immunoconjugates
• Oligopeptides
• auristatin
• monomethylauristatin F
• Phenylalanine

Topics

• Aminobenzoates (chemistry)
• Animals
• Antineoplastic Agents (chemistry)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Cytotoxins (chemistry)
• Halogenation
• Humans
• Immunoconjugates (chemistry)
• Isomerism
• Magnetic Resonance Spectroscopy (methods)
• Mice
• Molecular Conformation
• Neoplasms (metabolism, pathology)
• Oligopeptides (chemistry)
• Phenylalanine (chemistry)