Synthesis and Evaluation of Hydrophilic Linkers for Antibody–Maytansinoid Conjugates

Zhao, Ruike Renee; Wilhelm, Sharon; Audette, Charlene A.; Jones, Gregory E.; Leece, Barbara; Lazar, Alexandru C.; Goldmacher, Victor S.; Singh, Rajeeva; Kovtun, Yelena; Widdison, Wayne C.; Lambert, John M.; Chari, Ravi

doi:10.1021/jm2002958

Cited by 180 publications

(164 citation statements)

References 41 publications

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“…1A and Scheme S1). The linker contains two ethylene glycol moieties modified with a terminal alkoxy-amine group to improve solubility and minimize export of the more hydrophilic derivatized analogs by drug pumps (32).…”

Section: Resultsmentioning

confidence: 99%

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

Axup

Bajjuri²,

Ritland³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

506

454

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Antibody-drug conjugates (ADCs) allow selective targeting of cytotoxic drugs to cancer cells presenting tumor-associated surface markers, thereby minimizing systemic toxicity. Traditionally, the drug is conjugated nonselectively to cysteine or lysine residues in the antibody. However, these strategies often lead to heterogeneous products, which make optimization of the biological, physical, and pharmacological properties of an ADC challenging. Here we demonstrate the use of genetically encoded unnatural amino acids with orthogonal chemical reactivity to synthesize homogeneous ADCs with precise control of conjugation site and stoichiometry. p -Acetylphenylalanine was site-specifically incorporated into an anti-Her2 antibody Fab fragment and full-length IgG in Escherichia coli and mammalian cells, respectively. The mutant protein was selectively and efficiently conjugated to an auristatin derivative through a stable oxime linkage. The resulting conjugates demonstrated excellent pharmacokinetics, potent in vitro cytotoxic activity against Her2 + cancer cells, and complete tumor regression in rodent xenograft treatment models. The synthesis and characterization of homogeneous ADCs with medicinal chemistry-like control over macromolecular structure should facilitate the optimization of ADCs for a host of therapeutic uses.

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Section: Resultsmentioning

confidence: 99%

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

Axup

Bajjuri²,

Ritland³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

506

454

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“…-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4) via the succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), the N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP), the N-succinimidyl 4-(2-pyridyldithio)butanoate (SPDB), or the N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB) linkers according to the protocols developed at ImmunoGen, Inc. and described previously (31)(32)(33). The maytansinoid per antibody ratio varied from 3.3 to 5.0.…”

Section: Cell Linesmentioning

confidence: 99%

IMGN853, a Folate Receptor-α (FRα)–Targeting Antibody–Drug Conjugate, Exhibits Potent Targeted Antitumor Activity against FRα-Expressing Tumors

Whiteman

Bartle

et al. 2015

Molecular Cancer Therapeutics

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A majority of ovarian and non-small cell lung adenocarcinoma cancers overexpress folate receptor a (FRa). Here, we report the development of an anti-FRa antibody-drug conjugate (ADC), consisting of a FRa-binding antibody attached to a highly potent maytansinoid that induces cell-cycle arrest and cell death by targeting microtubules. From screening a large panel of anti-FRa monoclonal antibodies, we selected the humanized antibody M9346A as the best antibody for targeted delivery of a maytansinoid payload into FRa-positive cells. We compared M9346A conjugates with various linker/maytansinoid combinations, and found that a conjugate, now denoted as IMGN853, with the N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB) linker and N

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“…Humanized anti-folate receptor-a (FRa), anti-EGFR, and anti-CD33 antibodies, chimeric mouse-human anti-EpCAM antibody, and chimeric non-binding control antibody to the Kunitz soybean protease inhibitor (KTI) were all constructed with the human IgG 1 isotype and generated at ImmunoGen, Inc. None of the antibodies used in the studies cross-react with antigen on mouse tissues. The antibody modifying agents SPDB [N-succinimidyl 4-(2-pyridyldithio)butanoate] and sulfo-SPDB [N-succinimidyl 2-sulfo-4-(2-pyridyldithio) butanoate] were prepared as previously described (20,21). Additional chemical reagents were purchased from Sigma-Aldrich.…”

Section: Cell Lines Antibodies and Reagentsmentioning

confidence: 99%

“…We have previously shown that incorporation of a sulfonate group in the disulfide linker confers improved water solubility to maytansinoid ADCs and also enhances antitumor efficacy in vivo (21,29). By analogy, we incorporated the charged sulfonate group in the IGN ADCs as part of the linker optimization effort.…”

Section: Linker Optimization and In Vivo Antitumor Activitymentioning

confidence: 99%

A New Class of Antibody–Drug Conjugates with Potent DNA Alkylating Activity

Miller

Fishkin

et al. 2016

Molecular Cancer Therapeutics

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The promise of tumor-selective delivery of cytotoxic agents in the form of antibody-drug conjugates (ADC) has now been realized, evidenced by the approval of two ADCs, both of which incorporate highly cytotoxic tubulin-interacting agents, for cancer therapy. An ongoing challenge remains in identifying potent agents with alternative mechanisms of cell killing that can provide ADCs with high therapeutic indices and favorable tolerability. Here, we describe the development of a new class of potent DNA alkylating agents that meets these objectives. Through chemical design, we changed the mechanism of action of our novel DNA cross-linking agent to a monofunctional DNA alkylator. This modification, coupled with linker optimization, generated ADCs that were well tolerated in mice and demonstrated robust antitumor activity in multiple tumor models at doses 1.5% to 3.5% of maximally tolerated levels. These properties underscore the considerable potential of these purpose-created, unique DNAinteracting conjugates for broadening the clinical application of ADC technology. Mol Cancer Ther; 15(8); 1870-8. Ó2016 AACR.

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Synthesis and Evaluation of Hydrophilic Linkers for Antibody–Maytansinoid Conjugates

Cited by 180 publications

References 41 publications

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

Synthesis of site-specific antibody-drug conjugates using unnatural amino acids

IMGN853, a Folate Receptor-α (FRα)–Targeting Antibody–Drug Conjugate, Exhibits Potent Targeted Antitumor Activity against FRα-Expressing Tumors

A New Class of Antibody–Drug Conjugates with Potent DNA Alkylating Activity

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