Impact of Drug Conjugation on Thermal and Metabolic Stabilities of Aglycosylated and <i>N</i>-Glycosylated Antibodies

Yamazoe, Sayumi; Kotapati, Srikanth; Hogan, Jason M.; West, Sean M.; Deng, Xiaolong; Diong, S J; Arbanas, Jaren; Nguyen, Thien Anh; Jashnani, Aarti; Gupta, Diksha; Rajpal, Arvind; Dollinger, Gavin; Strop, Pavel

doi:10.1021/acs.bioconjchem.1c00572

Cited by 6 publications

(4 citation statements)

References 32 publications

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“…No detectable decrease in T m and T agg was observed after MMAE conjugation while, unexpectedly, an increase of T m for approximately 10 °C was found in the human IgG-MMAE ADC. It has been reported that payload conjugation to the IgG CH2 domain could result in either positive and negative effects on T m , depending on the payload properties and the conjugation sites . MMAE molecules have relatively high hydrophobicity and likely adopt an inward orientation in the Fc region to minimize exposure to the hydrophilic surroundings.…”

Section: Resultsmentioning

confidence: 99%

“…It has been reported that payload conjugation to the IgG CH2 domain could result in either positive and negative effects on T m , depending on the payload properties and the conjugation sites. 60 MMAE molecules have relatively high hydrophobicity and likely adopt an inward orientation in the Fc region to minimize exposure to the hydrophilic surroundings. They could form nonpolar interactions with each other or with the hydrophobic IgG residues in the CH2 domain (e.g., F241, F243, V262, V264), which stabilizes the Fc conformation and results in a T m increase.…”

Section: Bioconjugate Chemistrymentioning

confidence: 99%

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Site-Specific Antibody Conjugation Using Modified Bisected N-Glycans: Method Development and Potential toward Tunable Effector Function

Hsu,

Nourzaie,

Tocher

et al. 2023

Bioconjugate Chem.

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Antibody−drug conjugates (ADCs) have garnered worldwide attention for disease treatment, as they possess high target specificity, a long half-life, and outstanding potency to kill or modulate the functions of targets. FDA approval of multiple ADCs for cancer therapy has generated a strong desire for novel conjugation strategies with high biocompatibility and controllable bioproperties. Herein, we present a bisecting glycan-bridged conjugation strategy that enables site-specific conjugation without the need for the oligosaccharide synthesis and genetic engineering of antibodies. Application of this method is demonstrated by conjugation of anti-HER2 human and mouse IgGs with a cytotoxic drug, monomethyl auristatin E. The glycan bridge showed outstanding stability, and the resulting ADCs eliminated HER2-expressing cancer cells effectively. Moreover, our strategy preserves the feasibility of glycan structure remodeling to fine-tune the immunogenicity and pharmacokinetic properties of ADCs through glycoengineering.

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Section: Resultsmentioning

confidence: 99%

Section: Bioconjugate Chemistrymentioning

confidence: 99%

Site-Specific Antibody Conjugation Using Modified Bisected N-Glycans: Method Development and Potential toward Tunable Effector Function

Hsu,

Nourzaie,

Tocher

et al. 2023

Bioconjugate Chem.

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“…Aglycosylation could be especially advantageous when the priority is to minimize the incidence of liver toxicities and inflammatory responses over enhancing potency. Aglycosylated antibodies can be vulnerable to structural distortion owing to thermal instability; nonetheless, data published in 2022 indicate that attaching small-molecule payloads to the CH 2 domain of the Fc region can compensate for this instability 15 .…”

Section: Review Articlementioning

confidence: 99%

Exploring the next generation of antibody–drug conjugates

Tsuchikama,

Anami,

et al. 2024

Nat Rev Clin Oncol

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Antibody-drug conjugates (ADCs) are a promising cancer treatment modality that enables the selective delivery of highly cytotoxic payloads to tumours. However, realizing the full potential of this platform necessitates innovative molecular designs to tackle several clinical challenges such as drug resistance, tumour heterogeneity and treatment-related adverse effects. Several emerging ADC formats exist, including bispecific ADCs, conditionally active ADCs (also known as probody-drug conjugates), immune-stimulating ADCs, protein-degrader ADCs and dual-drug ADCs, and each offers unique capabilities for tackling these various challenges. For example, probody-drug conjugates can enhance tumour specificity, whereas bispecific ADCs and dual-drug ADCs can address resistance and heterogeneity with enhanced activity. The incorporation of immune-stimulating and protein-degrader ADCs, which have distinct mechanisms of action, into existing treatment strategies could enable multimodal cancer treatment. Despite the promising outlook, the importance of patient stratification and biomarker identification cannot be overstated for these emerging ADCs, as these factors are crucial to identify patients who are most likely to derive benefit. As we continue to deepen our understanding of tumour biology and refine ADC design, we will edge closer to developing truly effective and safe ADCs for patients with treatment-refractory cancers. In this Review, we highlight advances in each ADC component (the monoclonal antibody, payload, linker and conjugation chemistry) and provide more-detailed discussions on selected examples of emerging novel ADCs of each format, enabled by engineering of one or more of these components.• To address these challenges, several novel ADC formats have been developed, including bispecific ADCs, probody-drug conjugates, immune-stimulating ADCs, protein-degrader ADCs and dual-drug ADCs.• Probody-drug conjugates are expected to have improved tumour specificity, whereas bispecific ADCs and dual-drug ADCs have the potential to combat drug resistance and tumour heterogeneity.• Integrating immune-stimulating ADCs and protein-degrader ADCs with current treatment regimens might enable multimodal treatment, potentially through several distinct mechanisms of action.• Patient stratification and biomarker identification will be crucial to maximize the clinical benefits of these emerging ADCs.Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

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“…It was recently reported that the introduction of small-molecule drugs into Gln of an aglycosylated antibody by using microbial transglutaminase (mTG) can have an impact on the stability of ADC. The hydrophobic cytotoxin was able to compensate for thermal destabilization resulting from structural distortions due to antibody deglycosylation [ 21 ]. The site-specific conjugation of Q295 in deglycosylated antibody was also described by using the same transglutaminase with cystamine for thiolation [ 23 ].…”

Section: Overview Of Site-specific Antibody Conjugationmentioning

confidence: 99%

Site-Specific Antibody Conjugation with Payloads beyond Cytotoxins

Zhou

2023

Molecules

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As antibody–drug conjugates have become a very important modality for cancer therapy, many site-specific conjugation approaches have been developed for generating homogenous molecules. The selective antibody coupling is achieved through antibody engineering by introducing specific amino acid or unnatural amino acid residues, peptides, and glycans. In addition to the use of synthetic cytotoxins, these novel methods have been applied for the conjugation of other payloads, including non-cytotoxic compounds, proteins/peptides, glycans, lipids, and nucleic acids. The non-cytotoxic compounds include polyethylene glycol, antibiotics, protein degraders (PROTAC and LYTAC), immunomodulating agents, enzyme inhibitors and protein ligands. Different small proteins or peptides have been selectively conjugated through unnatural amino acid using click chemistry, engineered C-terminal formylglycine for oxime or click chemistry, or specific ligation or transpeptidation with or without enzymes. Although the antibody protamine peptide fusions have been extensively used for siRNA coupling during early studies, direct conjugations through engineered cysteine or lysine residues have been demonstrated later. These site-specific antibody conjugates containing these payloads other than cytotoxic compounds can be used in proof-of-concept studies and in developing new therapeutics for unmet medical needs.

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Impact of Drug Conjugation on Thermal and Metabolic Stabilities of Aglycosylated and N-Glycosylated Antibodies

Cited by 6 publications

References 32 publications

Site-Specific Antibody Conjugation Using Modified Bisected N-Glycans: Method Development and Potential toward Tunable Effector Function

Site-Specific Antibody Conjugation Using Modified Bisected N-Glycans: Method Development and Potential toward Tunable Effector Function

Exploring the next generation of antibody–drug conjugates

Site-Specific Antibody Conjugation with Payloads beyond Cytotoxins

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