Supporting information

Supporting information

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Aim: Aur0101 is a cytotoxic and small-molecule microtubule depolymerizing agent, and is the payload conjugated to antibody–drug conjugate PYX-201. Developing and validating a sensitive bioanalytical method to quantitate Aur0101 was novel and crucial in preclinical PYX-201 studies. Materials & methods: Reference standard Aur0101 and its stable isotope labelled internal standard Aur0101-d8 were used in this LC–MS/MS method. Results: This sensitive assay was validated at a lower limit of quantitation of 15 pg/ml and successfully applied to support preclinical rat and monkey toxicology studies. Preclinical plasma toxicokinetic parameters were presented. Conclusion: A sensitive and robust LC–MS/MS assay was validated for Aur0101 in rat and monkey plasma.

Aim: PYX-201 is a novel antibody–drug conjugate targeting the extra domain B splice variant offibronectin in the tumor microenvironment. Accurate quantification of PYX-201 is critical for PYX-201 pharmacokinetics profiling in preclinical studies. Materials & methods: ELISA was performed usingreference standard PYX-201, mouse monoclonal anti-monomethyl auristatin E antibody, mouse IgG1,mouse monoclonal anti-human IgG horseradish peroxidase and donkey anti-human IgG horseradishperoxidase. Results: This assay was validated at 50.0–10,000 ng/ml in rat dipotassium EDTA plasma and250–10,000 ng/ml in monkey dipotassium EDTA plasma. Conclusion: This is the first time for a PYX-201bioanalytical assay in any matrix to be reported.

Aim: PYX-201 is a novel antibody–drug conjugate targeting the extra domain B splice variant offibronectin in the tumor microenvironment. Accurate quantification of PYX-201 is critical for PYX-201 pharmacokinetics profiling in preclinical studies. Materials & methods: ELISA was performed usingreference standard PYX-201, mouse monoclonal anti-monomethyl auristatin E antibody, mouse IgG1,mouse monoclonal anti-human IgG horseradish peroxidase and donkey anti-human IgG horseradishperoxidase. Results: This assay was validated at 50.0–10,000 ng/ml in rat dipotassium EDTA plasma and250–10,000 ng/ml in monkey dipotassium EDTA plasma. Conclusion: This is the first time for a PYX-201bioanalytical assay in any matrix to be reported.