A new, triglycyl peptide linker for antibody-drug conjugates (ADCs) with improved targeted killing of cancer cells

Published on Jun 1, 2016in Molecular Cancer Therapeutics5.615
· DOI :10.1158/1535-7163.MCT-16-0021
Rajeeva Singh23
Estimated H-index: 23
(ImmunoGen, Inc.),
Yulius Setiady15
Estimated H-index: 15
(ImmunoGen, Inc.)
+ 19 AuthorsWayne C. Widdison15
Estimated H-index: 15
(ImmunoGen, Inc.)
Sources
Abstract
A triglycyl peptide linker (CX) was designed for use in antibody-drug conjugates (ADCs), aiming to provide efficient release and lysosomal efflux of cytotoxic catabolites within targeted cancer cells. ADCs comprising anti-epithelial cell adhesion molecule (anti-EpCAM) and anti-epidermal growth factor receptor (anti-EGFR) antibodies with maytansinoid payloads were prepared using CX or a non-cleavable SMCC linker (CX and SMCC ADCs). The in vitro cytotoxic activities of CX and SMCC ADCs were similar for several cancer cell lines, however the CX ADC was more active (5-100 fold lower IC50) than the SMCC ADC in other cell lines including a multi-drug resistant line. Both CX and SMCC ADCs showed comparable maximum tolerated doses and pharmacokinetics in CD-1 mice. In Calu-3 tumor xenografts, antitumor efficacy was observed with the anti-EpCAM CX ADC at a five-fold lower dose than the corresponding SMCC ADC in vivo. Similarly, the anti-EGFR CX ADC showed improved antitumor activity over the respective SMCC conjugate in HSC-2 and H1975 tumor models, however both exhibited similar activity against FaDu xenografts. Mechanistically, in contrast to the charged lysine-linked catabolite of SMCC ADC, a significant fraction of the carboxylic acid catabolite of CX ADC could be uncharged in the acidic lysosomes and thus diffuse out readily into the cytosol. Upon release from tumor cells, CX catabolites are charged at extracellular pH and do not penetrate and kill neighboring cells, similar to the SMCC catabolite. Overall, these data suggest that CX represents a promising linker option for the development of ADCs with improved therapeutic properties.
Figures & Tables
Download
📖 Papers frequently viewed together
1 Citations
537 Citations
References35
Newest
#1Nikolaos Diamantis (ICR: Institute of Cancer Research)H-Index: 4
#2Udai Banerji (ICR: Institute of Cancer Research)H-Index: 43
Antibody-drug conjugates (ADCs) are an emerging novel class of anticancer treatment agents that combines the selectivity of targeted treatment with the cytotoxic potency of chemotherapy drugs. New linker technology associated with novel highly potent cytotoxic payloads has permitted the development of more effective and safe ADCs. In recent years, two ADCs have been licensed, T-DM1 and brentuximab vedotin, and are already establishing their place in cancer treatment. A plethora of ADCs are being...
239 CitationsSource
#1Kevin J. Hamblett (Amgen)H-Index: 13
#2Allison P. Jacob (Amgen)H-Index: 6
Last. William C. Fanslow (Amgen)H-Index: 55
view all 17 authors...
Antibody–drug conjugates (ADC) target cytotoxic drugs to antigen-positive cells for treating cancer. After internalization, ADCs with noncleavable linkers are catabolized to amino acid-linker-warheads within the lysosome, which then enter the cytoplasm by an unknown mechanism. We hypothesized that a lysosomal transporter was responsible for delivering noncleavable ADC catabolites into the cytoplasm. To identify candidate transporters, we performed a phenotypic shRNA screen with an anti-CD70 mayt...
47 CitationsSource
#1Wayne C. Widdison (ImmunoGen, Inc.)H-Index: 15
#2Jose F. Ponte (ImmunoGen, Inc.)H-Index: 10
Last. Ravi V. J. ChariH-Index: 30
view all 21 authors...
Antibody anilino maytansinoid conjugates (AaMCs) have been prepared in which a maytansinoid bearing an aniline group was linked through the aniline amine to a dipeptide, which in turn was covalently attached to a desired monoclonal antibody. Several such conjugates were prepared utilizing different dipeptides in the linkage including Gly-Gly, l-Val-l-Cit, and all four stereoisomers of the Ala-Ala dipeptide. The properties of AaMCs could be altered by the choice of dipeptide in the linker. Each o...
16 CitationsSource
#1Paulin Salomon (ImmunoGen, Inc.)H-Index: 6
#2Rajeeva Singh (ImmunoGen, Inc.)H-Index: 23
A new, sensitive ELISA method has been developed which measures catabolites in cells and media upon processing of antibody–drug conjugates (ADCs) by target cancer cells. This ELISA method, exemplified for maytansinoid ADCs, uses competitive inhibition by a maytansinoid analyte of the binding of biotinylated antimaytansine antibody to an immobilized BSA–maytansinoid conjugate. Synthetic standards of several maytansinoid catabolites derived from ADCs with different linkers were tested and showed s...
19 CitationsSource
Antibody–drug conjugates (ADCs) that are currently on the market or in clinical trials are predominantly based on two drug classes: auristatins and maytansinoids. Both are tubulin binders and block the cell in its progression through mitosis. We set out to develop a new class of linker-drugs based on duocarmycins, potent DNA-alkylating agents that are composed of a DNA-alkylating and a DNA-binding moiety and that bind into the minor groove of DNA. Linker-drugs were evaluated as ADCs by conjugati...
97 CitationsSource
Ado-trastuzumab emtansine (T-DM1) is an antibody–drug conjugate that combines the antitumor properties of the humanized anti-human epidermal growth factor receptor 2 (HER2) antibody, trastuzumab, with the maytansinoid, DM1, a potent microtubule-disrupting agent, joined by a stable linker. Upon binding to HER2, the conjugate is internalized via receptor-mediated endocytosis, and an active derivative of DM1 is subsequently released by proteolytic degradation of the antibody moiety within the lysos...
246 CitationsSource
#1Ravi V. J. Chari (ImmunoGen, Inc.)H-Index: 30
#2Michael L. Miller (ImmunoGen, Inc.)H-Index: 17
Last. Wayne C. Widdison (ImmunoGen, Inc.)H-Index: 15
view all 3 authors...
Traditional cancer chemotherapy is often accompanied by systemic toxicity to the patient. Monoclonal antibodies against antigens on cancer cells offer an alternative tumor-selective treatment approach. However, most monoclonal antibodies are not sufficiently potent to be therapeutically active on their own. Antibody–drug conjugates (ADCs) use antibodies to deliver a potent cytotoxic compound selectively to tumor cells, thus improving the therapeutic index of chemotherapeutic agents. The recent a...
537 CitationsSource
#1May Kung Sutherland (Seattle Genetics)H-Index: 16
#2Roland B. Walter (Fred Hutchinson Cancer Research Center)H-Index: 60
Last. Julie A. McEarchern (Seattle Genetics)H-Index: 11
view all 20 authors...
Outcomes in acute myeloid leukemia (AML) remain unsatisfactory, and novel treatments are urgently needed. One strategy explores antibodies and their drug conjugates, particularly those targeting CD33. Emerging data with gemtuzumab ozogamicin (GO) demonstrate target validity and activity in some
292 CitationsSource
#1Kristi Elkins (Genentech)H-Index: 11
#2Bing ZhengH-Index: 14
Last. Andrew PolsonH-Index: 21
view all 13 authors...
Fc receptor-like 5 (FcRL5/FcRH5/IRTA2/CD307) is a surface protein expressed selectively on B cells and plasma cells. We found that FcRL5 was expressed at elevated levels on the surface of plasma cells from the bone marrow of patients diagnosed with multiple myeloma. This prevalence in multiple myeloma and narrow pattern of normal expression indicate that FcRL5 could be a target for antibody-based therapies for multiple myeloma, particularly antibody–drug conjugates (ADC), potent cytotoxic drugs ...
71 CitationsSource
#1Hans K. Erickson (ImmunoGen, Inc.)H-Index: 17
#2Gail Lewis PhillipsH-Index: 26
Last. Jay TibbittsH-Index: 13
view all 11 authors...
Trastuzumab emtansine (T-DM1) is an antibody–drug conjugate consisting of the anti-HER2 antibody trastuzumab linked via a nonreducible thioether linker to the maytansinoid antitubulin agent DM1. T-DM1 has shown favorable safety and efficacy in patients with HER2-positive metastatic breast cancer. In previous animal studies, T-DM1 exhibited better pharmacokinetics (PK) and slightly more efficacy than several disulfide-linked versions. The efficacy findings are unique, as other disulfide-linked an...
144 CitationsSource
Cited By15
Newest
#1Zheng Su (SPU: Shenyang Pharmaceutical University)
#2Xiao DianH-Index: 1
Last. Li Song (SPU: Shenyang Pharmaceutical University)H-Index: 1
view all 8 authors...
Abstract Antibody–drug conjugates (ADCs) are gradually revolutionizing clinical cancer therapy. The antibody‒drug conjugate linker molecule determines both the efficacy and the adverse effects, and so has a major influence on the fate of ADCs. An ideal linker should be stable in the circulatory system and release the cytotoxic payload specifically in the tumor. However, existing linkers often release payloads nonspecifically and inevitably lead to off-target toxicity. This defect is becoming an ...
Source
#1Diogo FigueiredoH-Index: 1
#2Célia FernandesH-Index: 13
Last. António PauloH-Index: 29
view all 8 authors...
For effective Auger therapy of cancer, the Auger-electron emitters must be delivered to the tumor cells in close proximity to a radiosensitive cellular target. Nuclear DNA is considered the most relevant target of Auger electrons to have augmented radiotoxic effects and significant cell death. However, there is a growing body of evidence that other targets, such as the mitochondria, could be relevant subcellular targets in Auger therapy. Thus, we developed dual-targeted 99mTc(I) tricarbonyl comp...
1 CitationsSource
#1Lu Liu (MHH: Hannover Medical School)H-Index: 1
#2Jürgen Borlak (MHH: Hannover Medical School)H-Index: 23
Over the last decade research on cancer stem cells (CSC) significantly contributed to a better understanding of tumor biology. Given their similarity to normal stem cells, i.e. self-renewal and pluripotency the need arises to develop robust protocols for the isolation and characterization of CSCs. As with other malignancies, hepatic tumors are composed of a heterogeneous population of cells including liver cancer stem cells (LCSC). Yet, a precise understanding of why stem cells become cancerous ...
1 CitationsSource
#1Fiona Concy Rodrigues (MIT: Manipal Institute of Technology)
#2N. G. Devi (MIT: Manipal Institute of Technology)H-Index: 1
Last. Goutam Thakur (MIT: Manipal Institute of Technology)H-Index: 10
view all 3 authors...
Abstract Cancer, the abnormal proliferation of cells, is one of the leading causes of death in the world. A number of challenges hamper its treatment regime, such as ineffective drug concentration reaching the target site, nonspecific distribution of drug and acquired resistance upon chemotherapy due to various drugs and dosage. With a deeper understanding of tumor biology and the advancement in pharmaceutical technology, drug delivery vehicles have been recognized as a suitable approach in its ...
Source
#1Puregmaa Khongorzul (CPU: China Pharmaceutical University)H-Index: 3
#2Cai Jia Ling (CPU: China Pharmaceutical University)H-Index: 1
Last. Juan Zhang (CPU: China Pharmaceutical University)H-Index: 1
view all 5 authors...
Antibody-drug conjugates (ADCs) are one of the fastest-growing anti-cancer drugs. This approach comprises a monoclonal antibody conjugated to the cytotoxic payload via a chemical linker that directed towards a target antigen expressed on the cancer cell surface, reducing systemic exposure and therefore toxicity. ADCs are complex molecules that require careful attention to various components. Selection of an appropriate target, a monoclonal antibody, cytotoxic payload, and the manner in which the...
68 CitationsSource
The prototypical ADC mechanism involving antigen-mediated uptake and lysosomal release is both elegantly simple and scientifically compelling. However, recent clinical-stage failures have prompted a reevaluation of this delivery paradigm and have resulted in an array of new technologies that have the potential to improve the safety and efficacy of up and coming programs. These innovations can generally be categorized into seven areas that will be elaborated on in this chapter: (1) Exploiting new...
1 CitationsSource
#1Chuan Shi (Janssen Pharmaceutica)H-Index: 3
#2Shalom Goldberg (Janssen Pharmaceutica)H-Index: 10
Last. Wenying Jian (Janssen Pharmaceutica)H-Index: 1
view all 12 authors...
Aim: Alternative scaffold proteins have emerged as novel platforms for development of therapeutic applications. One such application is in protein–drug conjugates (PDCs), which are analogous to ant...
6 CitationsSource
#1Jose M. Pacheco (University of Colorado Denver)H-Index: 12
#2D. Ross Camidge (University of Colorado Denver)H-Index: 83
Abstract Antibody drug conjugates (ADCs) have the potential to alter the efficacy: toxicity ratio of cytotoxic therapy utilizing surface markers on cancer cells as antibody targets to preferentially deliver toxic payloads to tumor cells while limiting systemic toxicity. Multiple ADCs, differing in their antibody targets, cytotoxic payloads and linker molecules are currently being evaluated in non-small-cell lung cancer, small-cell lung cancer and malignant pleural mesothelioma. Here we review th...
6 CitationsSource
The antibody–drug conjugate (ADC) field is in a transitional period. Older approaches to conjugate composition and dosing regimens still dominate the ADC clinical pipeline, but preclinical work is driving a rapid evolution in how we strategize to improve efficacy and reduce toxicity towards better therapeutic outcomes. These advances are largely based upon a body of investigational studies that together offer a deeper understanding of the absorption, distribution, metabolism, and excretion (ADME...
36 CitationsSource
#1Yiwu ZhengH-Index: 5
#2Jing RenH-Index: 6
Last. Chuanliu WuH-Index: 18
view all 6 authors...
Targeted prodrugs exploiting cleavable linkers capable of responding to endogenous stimuli have increasingly been explored for cancer therapy. Successful application of these prodrug designs relies on the manipulation of both stability and responsiveness of the cleavable linkers, which, however, are difficult to be finely regulated, particularly for acid-responsive acylhydrazone bonds. Here we developed a new class of peptide-bridged twin-acylhydrazone linkers (PTA linkers) displaying both an ul...
8 CitationsSource