Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates

Lyon, Robert P.; Setter, Jocelyn R.; Bovee, Tim D.; Doronina, Svetlana O.; Hunter, Joshua H.; Anderson, Martha E.; Balasubramanian, Cindy; Duniho, Steven; Leiske, Chris; Li, Fu; Senter, Peter D.

doi:10.1038/nbt.2968

Cited by 367 publications

(365 citation statements)

References 28 publications

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“…It has previously been reported that replacement of the MC stretcher with mDPR has no effect on in vitro ADC potency and activity, but that the improved stability may lead to increased in vivo potency in a model-dependent manner (15). On the basis of these results, the impact of mDPR inclusion on intratumoral drug delivery was assessed.…”

Section: Mdpr Inclusion Increases Mmae Delivery In Vivo and Stabilitymentioning

confidence: 99%

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Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody–Drug Conjugates

Burke

Hamilton

Jeffrey

et al. 2017

Molecular Cancer Therapeutics

116

111

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The emergence of antibody-drug conjugates (ADC), such as brentuximab vedotin and ado-trastuzumab emtansine, has led to increased efforts to identify new payloads and develop improved drug-linker technologies. Most antibody payloads impart significant hydrophobicity to the ADC, resulting in accelerated plasma clearance and suboptimal in vivo activity, particularly for conjugates with high drug-to-antibody ratios (DAR). We recently reported on the incorporation of a discrete PEG 24 polymer as a side chain in a b-glucuronidase-cleavable monomethylauristatin E (MMAE) linker to provide homogeneous DAR 8 conjugates with decreased plasma clearance and increased antitumor activity in xenograft models relative to a non-PEGylated control. In this work, we optimized the drug-linker by minimizing the size of the PEG side chain and incorporating a self-stabilizing maleimide to prevent payload de-conjugation in vivo. Multiple PEG-glucuronide-MMAE linkers were prepared with PEG size up to 24 ethylene oxide units, and homogeneous DAR 8 ADCs were evaluated. A clear relationship was observed between PEG length and conjugate pharmacology when tested in vivo. Longer PEG chains resulted in slower clearance, with a threshold length of PEG 8 beyond which clearance was not impacted. Conjugates bearing PEG of sufficient length to minimize plasma clearance provided a wider therapeutic window relative to faster clearing conjugates bearing shorter PEGs. A lead PEGylated glucuronide-MMAE linker was identified incorporating a self-stabilizing maleimide and a PEG 12 side chain emerged from these efforts, enabling highly potent, homogeneous DAR 8 conjugates and is under consideration for future ADC programs.

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Section: Mdpr Inclusion Increases Mmae Delivery In Vivo and Stabilitymentioning

confidence: 99%

“…The self-stabilizing maleimide (mDPR; Fig. 1) was developed to undergo rapid postconjugation thiosuccinimide hydrolysis, to provide circulation-stable ADCs with a wider therapeutic window (15).…”

Section: Introductionmentioning

confidence: 99%

Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody–Drug Conjugates

Burke

Hamilton

Jeffrey

et al. 2017

Molecular Cancer Therapeutics

116

111

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“…Using this approach, an ADC is produced that is homogeneous and bears an average of 16 total drugs, split evenly between the two drug linkers. The carrier utilizes two recent advancements for the construction of ADCs with improved pharmacological activity: a self‐stabilizing maleimide (mDPR) to minimize drug‐linker deconjugation in vivo,2a and a PEG 24 stretcher to enable high drug loading without concomitant hydrophobicity‐induced ADC aggregation 6…”

mentioning

confidence: 99%

“…B) A multiplexing drug carrier 4 bearing Cys(SiPr) and Cys(Acm) groups that can be unmasked using orthogonal conditions. The carrier also contains a PEG 24 group to mask drug‐linker hydrophicity6 and a self‐stabilizing maleimide (mDPR)2a for antibody attachment. C) Homogeneous dual‐drug ADCs prepared using 4 bear 16 total drugs, split evenly (8+8) between the two component drugs.…”

mentioning

confidence: 99%

Orthogonal Cysteine Protection Enables Homogeneous Multi‐Drug Antibody–Drug Conjugates

et al. 2016

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A strategy for the preparation of homogeneous antibody–drug conjugates (ADCs) containing multiple payloads has been developed. This approach utilizes sequential unmasking of cysteine residues with orthogonal protection to enable site‐specific conjugation of each drug. In addition, because the approach utilizes conjugation to native antibody cysteine residues, it is widely applicable and enables high drug loading for improved ADC potency. To highlight the benefits of ADC dual drug delivery, this strategy was applied to the preparation of ADCs containing two classes of auristatin drug‐linkers that have differing physiochemical properties and exert complementary anti‐cancer activities. Dual‐auristatin ADCs imparted activity in cell line and xenograft models that are refractory to ADCs comprised of the individual auristatin components. This work presents a facile method for construction of potent dual‐drug ADCs and demonstrates how delivery of multiple cytotoxic warheads can lead to improved ADC activities. Lastly, we anticipate that the conditions utilized herein for orthogonal cysteine unmasking are not restricted to ADCs and can be broadly utilized for site‐specific protein modification.

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“…This technology, when applied to trastuzumab emtansine, yields a defined DAR of 2, and the authors believe that the antitumor activity and stability is increased compared to drug conjugates produced with the THIOMAB method. A possible explanation might be that a retro-Michael addition, leading to the transfer of drugs conjugated to cysteines to free thiols in blood serum components, may have partially inactivated the ADC [76][77][78] . However, the authors do not provide convincing data showing that their own preparation of the THIOMAB antibody had a DAR of 2, as it was published by Junutula et al 50 .…”

Section: Click Chemistries For Bioconjugationmentioning

confidence: 99%

Antibody–Drug Conjugates for Tumor Targeting—Novel Conjugation Chemistries and the Promise of non-IgG Binding Proteins

et al. 2015

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Antibody-drug conjugates (ADCs) have emerged as a promising class of anti-cancer agents, combining the specificity of antibodies for tumor targeting and the destructive potential of highly potent drugs as payload. An essential component of these immunoconjugates is a bifunctional linker capable of reacting with the antibody and the payload to assemble a functional entity. Linker design is fundamental, as it must provide high stability in the circulation to prevent premature drug release, but be capable of releasing the active drug inside the target cell upon receptor-mediated endocytosis. Although ADCs have demonstrated an increased therapeutic window, compared to conventional chemotherapy in recent clinical trials, therapeutic success rates are still far from optimal. To explore other regimes of half-life variation and drug conjugation stoichiometries, it is necessary to investigate additional binding proteins which offer access to a wide range of formats, all with molecularly defined drug conjugation. Here, we delineate recent progress with site-specific and biorthogonal conjugation chemistries, and discuss alternative, biophysically more stable protein scaffolds like Designed Ankyrin Repeat Proteins (DARPins), which may provide such additional engineering opportunities for drug conjugates with improved pharmacological performance.3

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Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates

Cited by 367 publications

References 28 publications

Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody–Drug Conjugates

Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody–Drug Conjugates

Orthogonal Cysteine Protection Enables Homogeneous Multi‐Drug Antibody–Drug Conjugates

Antibody–Drug Conjugates for Tumor Targeting—Novel Conjugation Chemistries and the Promise of non-IgG Binding Proteins

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