Physical characteristics comparison between maytansinoid-based and auristatin-based antibody-drug conjugates

Abstract

Aim: Direct analytical comparison of two major drug-linkers in the antibody-drug conjugate (ADC) field was conducted. Methods: Four different analytical methods [AlogP calculation, reverse phase (RP) high-performance liquid chromatography (HPLC; RP-HPLC), size exclusion chromatography HPLC (SEC-HPLC), and differential scanning calorimetry (DSC)] were tested for this comparison. Results: Maytansinoid-based ADCs showed less hydrophobicity than auristatin-based ADCs. Regardless of the drug-linker and drug-to-antibody ratios (DARs), the stability detected by DSC was decreased by conjugation. Conclusions: The cost and time-efficient analytical comparison described in this manuscript may be useful information for an initial characterization of ADCs prior to detailed biological studies.

Enzymatic glycan remodeling–metal free click (GlycoConnect™) provides homogenous antibody-drug conjugates with improved stability and therapeutic index without sequence engineering

“…The resulting GlycoConnect™ ADCs obtained from trastuzumab- 1 / 3 conjugated to 4 or from brentuximab- 1 / 3 conjugated to 5 were assessed head-to-head for in vitro stability with regards to aggregation (pH 5, 40°C). The enhanced aggregation levels of DAR4 ADCs based on 5 (red lines in Figure 4b ) versus the DAR2 ADCs based 4 (blue lines) were expected, 27 given the two-fold higher number of lipophilic payloads. However, it was surprisingly found that under these forcing conditions both ADCs obtained by remodeling with azidosugar 6-azidoGalNAc ( 3 ) showed reduced aggregation versus analogues ADCs remodeled with GalNAz ( 1 ), irrespective of whether they were conjugated to 4 or 5 .…”

Tag‐Free Enzymatic Modification for Antibody−Drug Conjugate Production

“…ADCs produced using MTG‐mediated conjugation showed significant stability in mouse serum, and in vitro cytotoxicity and xenograft studies showed that these ADCs were sufficiently efficacious. Furthermore, thermal stability studies using differential scanning calorimetry [74] showed very similar values between the ADC and native mAb, alleviating previous concerns that the pharmaceutical development of transglutaminase/nonglycosylated mAb conjugates was not feasible due to low thermal stability.…”

Biological Evaluation of Maytansinoid-Based Site-Specific Antibody-Drug Conjugate Produced by Fully Chemical Conjugation Approach: AJICAP®

“…2C). Several previous studies, including ours, had reported stability assessments results of T-DM1 in peer-reviewed literature; and we followed the same procedures for one of the processes applied to trastuzumab-AJICAP-maytansinoid [13,[16][17][18].…”

“…To adapt the AJI-CAP method of conjugation, a cysteine-reactive maytansinoid (MCC-maytansinoid, Fig. 1) was synthesized as previously reported [16]. The stability of trastuzumab-AJICAPmaytansinoid was assessed to determine its appropriate storage conditions [17,18].…”