Stabilization of cysteine-linked antibody drug conjugates with N-aryl maleimides

Christie, R. James; Fleming, Ryan; Bezabeh, Binyam; Woods, R. Jeremy; Mao, Shenlan; Harper, Jay; Joseph, Augustine R.; Wang, Qianli; Xu, Zhenghu; Wu, Herren; Gao, Changshou; Dimasi, Nazzareno

doi:10.1016/j.jconrel.2015.09.032

Cited by 96 publications

(98 citation statements)

References 36 publications

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“…N ‐aryl maleimides arose as novel Michael acceptors capable of yielding more stable thiol‐based bioconjugates. Dimasi and co‐workers anticipated that replacing a N ‐alkyl maleimide substituent with an aromatic ring would increase the imide electrophilicity and, consequently, accelerate thiosuccinimide hydrolysis (Scheme A) . The authors synthesized three maleimides featuring distinct electrophilic levels, N ‐alkyl 5 , N ‐phenyl 6 , and N ‐(4‐fluorophenyl) 7 , and studied their ability to yield stable bioconjugates with monoclonal antibody T289C.…”

Section: Maleimide Chemistry In Bioconjugationmentioning

confidence: 99%

“…As observed previously, both aryl substitution and close proximity to electron‐withdrawing groups (e.g., amides) led to a considerable acceleration on the hydrolysis rate of dithiomaleimides (as indicated by hydrolysis half‐life) (Schemes B,C). In fact, the measured rate is comparable to the one observed for N ‐aryl thiosuccinimides . By affecting the hydrolytic stability of the bioconjugates, the stability of the dibromomaleimide reagent can become an important liability.…”

Section: Maleimide Chemistry In Bioconjugationmentioning

confidence: 99%

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Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

Ravasco

Faustino

Trindade

et al. 2018

Chemistry A European J

341

329

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Maleimide chemistry stands out in the bioconjugation toolbox by virtue of its synthetic accessibility, excellent reactivity, and practicability. The second-generation of clinically approved antibody-drug conjugates (ADC) and much of the current ADC pipeline in clinical trials contain the maleimide linkage. However, thiosuccinimide linkages are now known to be less robust than once thought, and ergo, are correlated with suboptimal pharmacodynamics, pharmacokinetics, and safety profiles in some ADC constructs. Rational design of novel generations of maleimides and maleimide-type reagents have been reported to address the shortcomings of classical maleimides, allowing for the formation of robust bioconjugate linkages. This review highlights the main strategies for rational reagent design that have allowed irreversible bioconjugations in cysteines, reversible labelling strategies and disulfide re-bridging.

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Section: Maleimide Chemistry In Bioconjugationmentioning

confidence: 99%

Section: Maleimide Chemistry In Bioconjugationmentioning

confidence: 99%

Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

Ravasco

Faustino

Trindade

et al. 2018

Chemistry A European J

341

329

View full text Add to dashboard Cite

show abstract

“…10,41,48 N-Alkyl succinimide conjugates can be stabilised against retro-Michael reactions by hydrolysis to succinamic acid derivatives, [49][50][51] which resulted in ADCs with improved pharmacokinetics and in vivo efficacy. 41,51,52 Most Nalkyl thiosuccinimide ADCs require prolonged hydrolysis (>16 h) at high pH (>8) 41,51 which is not desirable from a process view and can lead to deamidation and loss of payload.…”

Section: 35-39mentioning

confidence: 99%

Use of a next generation maleimide in combination with THIOMAB™ antibody technology delivers a highly stable, potent and near homogeneous THIOMAB™ antibody-drug conjugate (TDC)

et al. 2017

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Herein we demonstrate that conjugation of a next generation maleimide (NGM) to engineered cysteines in a THIOMAB™ antibody delivers a THIOMAB™ antibody-drug conjugate (TDC) with a drug loading of ca. 2. This TDC is highly stable in blood serum conditions, selective and potent towards HER2 expressing cell lines and meets the current criteria for optimised antibody-drug conjugates (ADCs).Antibody-drug conjugates (ADCs) have emerged as targeted therapeutics against cancer by combining the selectivity of the antibody component with the potency of cytotoxic drugs. There are two FDA-approved ADCs already on the market (Adcetris™ and Kadcyla™) and over 40 additional ADCs currently undergoing clinical trials. 1-4Classical methods of conjugation for producing ADCs include lysine modication 5 and cysteine modication via reduction of solvent accessible disulde bonds followed by reaction with maleimides.6 It has been recognised that both methods generate heterogeneous mixtures of ADCs with different drug loadings and varied sites of attachment, resulting in sub-optimal therapeutic indices and higher clearance rates.7-13 Formation of higher drug-to-antibody ratio species (DAR > 4) have narrower therapeutic indices. 1,7,9,12,14 The latter method also results in the loss of structural disulde bonds which can lead to poor stability. 7,9,14The next generation of conjugation methods has aimed to reduce heterogeneity by focusing on site-selectivity. Chemoenzymatic methods have been used for ADC synthesis such as glycan modication, 15-17 glutamine amidation 18 and cysteine oxidation to aldehyde tags.19 Site-selective disulde modica-tion with small molecules has been achieved using nextgeneration maleimides (NGMs), 20-27 pyridazinediones (PDs)28-32 and bis-sulfones. 33,34 Site-specic reactivity has also been accomplished through incorporation of non-natural aminoacids into proteins for bioorthogonal conjugation. 10,35-39The THIOMAB™ antibody technology platform uses sitedirected mutagenesis to incorporate cysteines into the antibody. Conjugation to a classical maleimide bearing the drug of choice affords a THIOMAB™ antibody-drug conjugate (TDC) with full control over site reactivity and a precise drug-toantibody ratio (DAR).12,40-42 The technology also avoids issues with some chemoenzymatic methods that can result in over oxidation of aminoacids 43 and aldehyde tag conversion to unreactive gem-diols.44 TDCs have been demonstrated to improve therapeutic indices and pharmacodynamics over ADCs prepared by classical cysteine and lysine modication. 11,12,42Maleimides are rapid thiol-selective conjugation reagents.45,46 However, the succinimide conjugates thus formed can undergo retro-Michael reactions and scavenging with albumin, 47 resulting in premature drug release during blood circulation.10,41,48 N-Alkyl succinimide conjugates can be stabilised against retro-Michael reactions by hydrolysis to succinamic acid derivatives, 49-51 which resulted in ADCs with improved pharmacokinetics and in vivo efficacy. 41,51,52 Most Nalkyl t...

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“…In this case, N-phenyl maleimide functionality was incorporated into the cytotoxic agent valinecitrulline- p -aminobenzyloxycarbonyl-monomethyl-auristatin-E (Val-Cit-PAB-MMAE) to permit delivery in response to GSH. 235 In general, while reduction-responsive peptide-polymer conjugates have great potential in intracellular drug delivery, most of the studies demonstrated the disulfide bond cleavage after in vitro treatment with DTT or GSH. The presence of free cysteine and homocysteine in human plasma offers an alternative trigger, but, unfortunately, this approach may result in the cleavage of the disulfide bond before it reaches the specific target, which could result in premature drug release.…”

Section: Responsive Peptide-mediated Assemblymentioning

confidence: 99%

Responsive hybrid (poly)peptide–polymer conjugates

et al. 2017

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(Poly)peptide-polymer conjugates continue to garner significant interest in the production of functional materials given their composition of natural and synthetic building blocks that confer select and synergistic properties. Owing to opportunities to design predefined architectures and structures with different morphologies, these hybrid conjugates enable new approaches for producing micro- or nanomaterials. Their modular design enables the incorporation of multiple responsive properties into a single conjugate. This review presents recent advances in (poly)peptide-polymer conjugates for drug-delivery applications, with a specific focus on the utility of the (poly)peptide component in the assembly of particles and nanogels, as well as the role of the peptide in triggered drug release.

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Stabilization of cysteine-linked antibody drug conjugates with N-aryl maleimides

Cited by 96 publications

References 36 publications

Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

Bioconjugation with Maleimides: A Useful Tool for Chemical Biology

Use of a next generation maleimide in combination with THIOMAB™ antibody technology delivers a highly stable, potent and near homogeneous THIOMAB™ antibody-drug conjugate (TDC)

Responsive hybrid (poly)peptide–polymer conjugates

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