Therapeutic advances have improved cancer survival outcomes for an increasing number of patients, but novel approaches are still urgently needed for patients who cannot tolerate, or do not respond to current treatments. Erythropoietin-producing hepatocellular receptor A2 (EphA2) is highly expressed in a variety of solid tumors, which is associated with poor prognosis, especially in tumors considered difficult-to-treat, such as pancreatic and head and neck cancer. EphA2 has emerged as a promising therapeutic target for the treatment of solid tumors; however, efficacy and safety issues have halted clinical development of previous EphA2-targeting agents including MEDI-547, DS-8895a, MM-310, and dasatinib. Despite these setbacks, interest in targeting EphA2 in solid tumors remains, with ongoing development of investigational therapies such as antibodies, antibody drug conjugates, EphA2 antagonists, peptide drug conjugates, bicyclic peptide drug conjugates, and tyrosine kinase inhibitors. Among these, BT5528, a Bicycle® Drug Conjugate (BDC), has shown an emerging differentiated safety profile, in contrast to prior EphA2-targeting agents, and promising antitumor activity in patients with advanced solid tumors. BT5528 comprises an EphA2-targeting bicyclic (Bicycle) peptide, linked to the cytotoxin monomethyl auristatin E (MMAE) via a valine-citrulline cleavable linker. The high specificity of BT5528 to EphA2, combined with its high affinity, enables precision-guided delivery of MMAE, while its peptidic nature results in rapid distribution and retention of MMAE within the tumor, limited systemic exposure, and liver-sparing renal elimination. The preclinical and emerging clinical data for BT5528 suggest that novel approaches to targeting EphA2 can achieve efficacy without the safety issues that plagued earlier agents. Here, we review EphA2 as a target and the historical and current clinical development of EphA2-targeting therapeutic agents.

Membrane type 1 matrix metalloproteinase (MT1-MMP) is a pivotal enzyme involved in extracellular matrix remodeling, contributing to tumor invasion, metastasis, and poor prognosis in various cancers, including non-small cell lung, urothelial, pancreatic, gastric, and breast cancers. This study outlines the preclinical development and first in-human application of a phage display-derived MT1-MMP-specific bicyclic peptide, [68Ga]Ga-BCY25286, as a radiotheranostic agent for PET/CT imaging. The MT1-MMP-targeting bicyclic peptide BCY25286 was radiolabeled with either Ga-68 or Lu-177 and subsequently characterized for stability, binding affinity, and internalization. Preclinical evaluation included biodistribution and μPET/MR imaging in MT1-MMP+ HT1080 and MT1-MMP- MCF-7 xenograft tumor-bearing nude mice. For clinical translation, a 65-year-old patient with advanced pulmonary adenocarcinoma underwent [18F]FDG-PET/CT followed by [68Ga]Ga-BCY25286 PET/CT imaging, with PET scans performed after 60 minutes for [18F]FDG and up to 60 minutes for [68Ga]Ga-BCY25286 (compassionate use). Radiolabeling achieved >99% radiochemical purity for both radionuclides. [68Ga]Ga-BCY25286 demonstrated highly MT1-MMP-specific binding (7.2 ± 1.6 nM), proteolytic stability up to 72 hours, and rapid background clearance, thereby enhancing imaging contrast within 30 minutes. In mice, the tracer demonstrated high tumor uptake (10.6 ± 1.1 %ID/g at 1 h p.i.) with persistence up to 24 hours. In the clinical case, [68Ga]Ga-BCY25286 PET/CT imaging revealed high uptake in both primary and biopsy-confirmed metastatic sites, corroborating the findings of [18F]FDG-PET. SUVmaxvalues were comparable for lymph node metastases but higher for bone metastases in MT1-MMP-PET compared to [18F]FDG-PET, with significant kidney retention due to renal excretion. This first-in-human application of MT1-MMP-targeting [68Ga]Ga-BCY25286 demonstrates the feasibility for visualization of MT-1-MMP-expressing primary tumors and metastases, which is in line with the preclinical findings. These initial clinical results support further investigation of [68Ga]Ga-BCY25286 as a diagnostic tool with potential to improve tumor characterization and patient management strategies in MT1-MMP-positive cancers. Ann-Christin Eder, Mohamed A. Omrane, Anusha R. Regupathy, Mohamed El Fakiri, Nils Steinacker, Lisa-Charlotte Domogalla, Christoph-Ferdinand Wielenberg, Michael Mix, Johanna Lahdenranta, Ben Blakeman, Francesca Wood, Philip Huxley, Gemma E. Mudd, Matthias Eder, Philipp T. Meyer, Martin T. Freitag. Development and clinical translation of a phage display derived MT1-MMP-specific bicyclic peptide for radiotheranostic applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4601.

The membrane-type 1 matrix metalloproteinase MT1-MMP (or MMP14) is a member of the type 1 transmembrane proteinase family, highly overexpressed in several solid tumors and associated with poor prognosis (e.g. non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma, breast cancer, gastric cancer, and osteosarcoma). Its expression profile makes this target amenable to novel precision targeting with a toxin drug conjugate across a broad spectrum of solid tumors. Bicycle® drug conjugates (BDC®) intrinsically possess several advantages over other targeted cytotoxic modalities such as antibody-drug conjugates (ADCs). This Bicycle® advantage includes improved kinetics of extravasation and tumor penetration, fast renal clearance, and a lack of engagement of Fc receptors. Targeting MT1-MMP with a BDC® molecule represents an attractive potential approach for targeted tumor therapy. Bicycle® binders for MT1-MMP were identified using a proprietary phage display peptide technology consisting of highly diverse phage libraries of Bicycle® molecules, then conjugated to cleavable linkers and toxins to form Bicycle® Drug Conjugates (BDC®). Tool molecules were identified, containing an MT1-MMP binding Bicycle® conjugated to the cytotoxic auristatin derivative maleimidocaprolyl-monomethyl auristatin E (MMAE) via a proteolytically cleavable valine-citrulline (Val-Cit) linker. The properties of tool molecules were evaluated using a range of in vitro and in vivo models assessing MT1-MMP binding and specificity/selectivity, internalisation, in vitro cytotoxicity, and in vivo antitumor activity using xenograft models (cell and patient derived). Toxicology studies were performed in Han Wistar rats and Cynomologous monkeys. In vitro, tool molecules bound to MT1-MMP with high affinity (KD ∼3nM), and demonstrated both, cross-reactivity to species orthologues of MT1-MMP and selectivity over related human MMP family members by surface plasmon resonance (SPR) measurements. A fluorescent version of the MT1-MMP binding Bicycle® bound to cell lines dependent on expression levels of MT1-MMP, effectively internalised, and the tool BDC® molecule (MMAE containing) demonstrated MT1-MMP dependent cytotoxicity in an in vitro cell model.In vivo, the tool BDC® molecule demonstrated the expected PK profile in pre-clinical species of a short half life, and was effective in MT1-MMP expressing cell line (CDX) xenograft models in both mice and rats, showing complete tumor regression at 3mg/kg weekly dosing. The molecule also showed antitumor activity in a panel of NSCLC patient-derived xenograft models (PDX). Finally, in vivo toxicology (2 doses 1 week apart) demonstrated a similar tolerated range to other clinical stage BDC® molecules. Targeting MT1-MMP with a BDC® molecule shows potent antitumour activity in a range of solid tumour xenograft models, with a focus on NSCLC, and represents an attractive opportunity to target this novel tumor antigen for therapeutic benefit in this indication. Steve B. Ludbrook, Stephen Walsh, Gemma Mudd, Johanna Lahdenranta, Philip Huxley, Luca Mascheroni, Maximillian Harman, Jennifer Bré, Inma Rioja, Michael Skynner, Gavin Bennett. Targeting the novel tumor antigen MT1-MMP with Bicycle® Drug Conjugates (BDC®) for the potential treatment of NSCLC and other solid tumor indications [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2025 Oct 22-26; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2025;24(10 Suppl):Abstract nr B026.

Erythropoietin-producing hepatocellular receptor A2 (EphA2), is a receptor tyrosine kinase involved in cell-cell interactions. It is known to be overexpressed in various tumors and is associated with poor prognosis. EphA2 has been proposed as a target for theranostic applications. Low molecular weight peptide-based scaffolds with low nanomolar affinities have been shown to be ideal in such applications. Bicyclic peptides have emerged as an alternative to traditional peptides for this purpose, offering affinities comparable to antibodies due to their constrained nature, along with high tissue penetration, and improved stability compared to linear counterparts. This study presents the development and comprehensive in vitroand in vivo preclinical evaluation of BCY18469, a novel EphA2-targeting bicyclic peptide-based radiotheranostic agent. Methods: The EphA2-targeting Bicycle (R) peptide BCY18469 was identified through phage-display and chemically optimized. BCY18469 was radiolabeled with Ga-68, Lu-177 and In-111. The physicochemical properties, binding affinity and internalization as well as specificity of the peptide were evaluated in vitro. In vivo PET/MR and SPECT/CT imaging studies were performed using [Ga-68]Ga-BCY18469 and [In-111]In-BCY18469, respectively,along with biodistribution of [Lu-177]Lu-BCY18469 up to 24 h post injection in HT1080- and PC-3- tumor bearing BALB/c nu/nu EphA2-overexpressing xenograft mouse models. Results: The EphA2-targeting bicyclic peptide BCY18469 showed high binding affinity toward human and mouse EphA2 (1.9 and 3.8 nM, respectively). BCY18469 specifically bound and internalized into EphA2-expressing HT1080 cells. Imaging studies showed high tumor enrichment at early time-points (SUV of 1.7 g/mL at 1 h p.i. and 1.2 g/mL at 2 h p.i. in PET/MRI, HT1080 xenograft) with tumor contrast as early as 5 min p.i. and kidney-mediated clearance. Biodistribution studies revealed high early tumor uptake (19.5 +/- 3.5 %ID/g at 1 h p.i., HT1080 xenograft) with SPECT/CT imaging further confirming these findings (5.7 +/- 1.5 %ID/g at 1 h p.i., PC-3 xenograft). Conclusion: BCY18469 demonstrated high affinity, specific targeting of EphA2, a favorable biodistribution profile, and clearance through renal pathways. These findings underscore the potentially important role of bicyclic peptides in advancing radiotheranostic approaches and encourage additional translational research.

Abstract Bicycle Therapeutics is developing a unique class of chemically synthesized medicines. Based on its proprietary bicycle peptide (Bicycle®) phage display platform, Bicycles are a unique class of highly constrained peptides, which have demonstrated utility in the targeted delivery of different classes of payloads (for example cytotoxic agents, radioisotopes, immune modulators) into tumors. Bicycles are currently being explored in the clinic as Bicycle Toxin Conjugates® (BTCs) for targeted delivery of cytotoxic payloads into tumors. BTCs consist of a bicyclic peptide that is conjugated to a cytotoxic payload via a cleavable linker, which allows payload release in the tumor microenvironment or within the tumor cell. BTCs were developed to address the shortcomings of antibody drug conjugates (ADCs) in several ways. First, the small size of BTCs (∼4 kDa) compared to large biologic entities such as monoclonal antibody (mAb)-based conjugates (∼150 kDa) allows rapid distribution to tissues and extensive tumor penetration, which enables rapid delivery of payload into the tumor. Second, the peptidic nature of BTCs results in relatively short, yet tunable, duration of systemic exposure and liver-sparing renal elimination. These properties limit the body’s exposure to payload and should therefore minimize damage to normal tissue. In this body of work, we used in vitro cytotoxicity and cell uptake assays and mouse and rat cell line derived xenograft models for 1) cytotoxicity and anti-tumor activity evaluation, and 2) Bicycle and toxin uptake and biodistribution evaluation. Here, we show that BTCs targeting a number of different tumor antigens can deliver toxins to tumor tissue producing durable responses in a range of preclinical in vivo models, spanning several solid tumor indications. Citation Format: Stephen J. Walsh, Johanna Lahdenranta, Philip Huxley, Gemma Mudd, Gavin Bennett, Amy Brown, Katerine an Rietschoten, Liuhong Chen, Heather Scott, Gabriella Ivanova-Berndt, Katarzyna Dzionek, Mike Rigby, Olga Burenkova, Phil Jeffrey, Paul Beswick, Michael Skynner, Nicholas Keen. Bicycle Toxin Conjugates®for the treatment of solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5807.

Natural killer (NK) cells have the unique potential to recognize and kill tumor cells independently of MHC-I presentation of antigens, as well as to secrete cytokines that engage adaptive anti-tumor immunity and the function of cytolytic T cells. We have discovered and characterized chemically synthetic, constrained bicyclic peptides that bind with high affinity and specificity to NKp46, an activating receptor expressed selectively on NK cells in the tumor microenvironment. Chemical coupling to other bicyclic peptides specific for the tumor antigens EphA2 or MT-1 created NKp46 agonists whose function was completely conditional on binding to the tumor antigen. These chemical conjugates effectively convert the tumor antigen into a kill me signal for NK cells. Not only did these newly created tumor-immune cell agonists (TICAs) direct potent and efficient killing of human tumor cells by primary human NK cells in vitro, but they also caused secretion of the pro-inflammatory cytokines TNFα and IFNγ. Importantly, the TICAs directed production of FLT3 ligand, an essential mitogen for conventional dendritic cells which are central to the development of anti-tumor immunity in cancer. We illustrate the TICA-directed interaction of NK cells with tumor cells using confocal microscopy and we show that TICAs enable sustained function over multiple rounds of killing. These novel tools are well positioned to harness the potential of NK cells in the treatment of cancer. ### Competing Interest Statement All authors were full-time employees of Bicycle Therapeutics at the time that the work was conducted, and some own stock or stock options in Bicycle Therapeutics.

Abstract Introduction: Clinical studies in cancer patients have validated CD137 agonism as an activator of the immune system to enable tumor rejection. We have demonstrated that small, chemically synthetic bicyclic peptides can drive tumor-localized agonism of CD137 and anti-tumor immunity in mouse models. Here, we report the next stage of our work - delving into the mechanism of action of these novel agents and extending our program to serve patients whose tumors express EphA2. Experimental Procedures: MultiOmyx™ hyperplexed immunofluorescence assay was used to evaluate the expression of CD137 and EphA2 in head and neck squamous cell carcinoma samples. Human PBMC/tumor cell co-culture assays were used to assess CD137 Bicycle TICA™ in vitro bioactivity and syngeneic mouse tumor models were used to evaluate CD137 Bicycle TICA anti-tumor activity as mono- or combination therapies. Pharmacodynamic activity was evaluated by transcriptional profiling using NanoString assays or single cell RNA sequencing. Summary of the Data: Studies utilizing a syngeneic mouse model and deep RNA sequencing of tumors from CD137 Bicycle TICA-treated mice implicated intratumoral dendritic cells in addition to T cells as early responders to localized CD137 agonism and potential contributors to the anti-tumor response. Furthermore, a mouse efficacy model demonstrated synergy with checkpoint inhibitor therapy. Multiplex imaging revealed that EphA2 and CD137 are co-expressed in human tumors of high unmet medical need and therefore, using our highly modular Bicycle platform, we discovered BT7455, a Bicycle TICA that engages EphA2 and CD137 with high affinity, resulting in potent EphA2-dependent activity in vitro and robust anti-tumor activity in vivo with intermittent dosing in mouse models. Gene expression profiling of tumors revealed that BT7455 led to increased production of cytokines and chemokines known to drive T cell infiltration, the extent of which differentiated it from both a checkpoint inhibitor and an anti-CD137 monoclonal antibody agonist. BT7455 was well tolerated in preclinical species and liver function tests indicated no evidence of hepatic toxicity. Statement of Conclusions: In summary, we have identified EphA2 as a promising target to pair with a CD137 agonist. In addition to CD8+ T cells, tumor-resident dendritic cells are a likely contributor to anti-tumor immunity following treatment with Bicycle® TICAs, supporting utility for Bicycle® tumor-targeted CD137 agonists in solid tumors beyond those that are highly T cell-infiltrated. We have advanced a clinical development candidate, BT7455, to realize this potential for patients with EphA2-expressing cancers. Citation Format: Johanna Lahdenranta, Kristen Hurov, Heather Cohen, Lia Luus, Cara Bray, Peter Brown, Anna Devlen, Carly Campbell, Matthew Gray, Mike Kelly, Gemma Mudd, Punit Upadhyaya, Sailaja Battula, Kelvin Zhang, Anne-Sophie Dugast, Kevin McDonnell, Phil Brandish, Nicholas Keen. Tumor-targeted activation of CD137 using Bicycles: New insights into mechanism of action and discovery of BT7455, a clinical candidate for the treatment of EphA2-expressing cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5301.

The RFamide family of peptides represents an important class of GPCR ligand neuropeptides covering a wide range of biological functions. While many analogues of the highly conserved C-terminal RFamide motif within this peptide class have been synthesized and their functional significance elucidated, additional exploration of the structure activity relationship is of value. We have developed a novel linker for solid phase peptide synthesis (SPPS) which is able to anchor amine functionalised compounds for further elaboration. The acid labile benzofuranone based amine (ALBA) linker (5-(3-aminopropylcarbamoyl)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]benzoic acid) is compatible with Fmoc based SPPS and has two cleavage modes. As a proof of concept, the ALBA linker was used to successfully synthesise a novel analogue of Kisspeptin 10, the natural ligand for GPCR54, whereby the natural RFamide motif was replaced with an RFamine. Biological evaluation of the amine-containing analogue revealed that the group is not compatible with receptor activation. image

Thymic stromal lymphopoietin (TSLP) is an epithelial-derived pro-inflammatory cytokine involved in the development of asthma and other atopic diseases. We used Bicycle Therapeutics' proprietary phage display platform to identify bicyclic peptides (Bicycles) with high affinity for TSLP, a target that is difficult to drug with conventional small molecules due to the extended protein–protein interactions it forms with both receptors. The hit series was shown to bind to TSLP in a hotspot, that is also used by IL-7Rα. Guided by the first X-ray crystal structure of a small peptide binding to TSLP and the identification of key metabolites, we were able to improve the proteolytic stability of this series in lung S9 fractions without sacrificing binding affinity. This resulted in the potent Bicycle 46 with nanomolar affinity to TSLP (KD = 13 nM), low plasma clearance of 6.4 mL/min/kg, and an effective half-life of 46 min after intravenous dosing to rats.

Background Toll-like receptor 3 (TLR3) is an intracellular pattern recognition receptor aimed at binding double stranded RNA, which leads to cellular activation and proinflammatory cytokine secretion. Modulation of TLR3 biology can have significant impact in oncology and autoimmunity indications. In immuno-oncology, TLR3 agonists have been deployed as adjuvants to activate immune cells such as type I conventional dendritic cells that can help initiate the adaptive immune response to the tumor. In autoimmunity, TLR3 antagonists have been aimed at reducing immune cell over-activation. Despite a potential broad utility, systemic dsRNA TLR3 agonists and existing antagonists have not demonstrated clinical success due to lack of targeting and toxicity resulting from systemic activation. Therefore, the need for new molecular approaches to influence the signaling of TLR3 is evident. Bicycle® peptides are small molecules that penetrate rapidly into tissues and solid tumors and have a short half-life compared to biologics, potentially reducing systemic toxicity due to reduced exposure time. Methods Using the Bicycle® phage display platform, we have identified several families of Bicycles that bind the TLR3 extracellular domain. Bicycles were evaluated for their ability to modulate TLR3 signaling using a TLR3-overexpressing reporter cell assay. Binding and agonism was also assessed in monocyte derived macrophages that endogenously express TLR3. Conclusions This system allowed us to more closely evaluate Bicycles' ability to bind and modulate TLR3 signaling through known pathways leading to production of pro-inflammatory cytokines.

The file contains supplemental details related to sequence information of the bicyclic peptides referred to in the text, binding data, organ distribution data, PK parameters, mouse and human plasma stability, in vivo metabolism of BCY-B2 in mice, cell binding and internalization data, additional PET imaging data. It also contains detailed methods related to protein expression, phage selection, peptide synthesis, affinity determination by fluorescence polarization and SPR, radiolabeling, internalization, confocal microscopy, plasma protein binding, plasma stability of radiolabeled compounds, µPET imaging, autoradiography, immunohistochemistry, and 6 references related to these methods. €¢ Supplementary Tables S1 - S14: o Table S1: Affinities and plasma stabilities of BCY-B and stabilized BCY-C. o Table S2: Sequence information and binding data of bicyclic peptides referred to in text. o Table S3: Surface plasmon resonance (SPR) data. o Table S4-6: Organ distribution data, BCY-B3/B4 o Table S7: PK parameters. o Table S8: Selectivity data of BCY-C towards other metalloproteinases. o Table S9-11: Organ distribution data- BCYC2/C4 o Table S12-14: Organ distribution data- BCYD1/D2 o Table S15: Organ distribution data- MAb €¢ Supplementary Figures S1-S6: o Figure S1: Comparative HT1080 cell binding and internalization of non-stabilized BCY-B3 and stabilized BCY-C2 o Figure S2: Organ distribution time course studies with active BCY-B3, and inactive BCY-B4 in HT1080 xenograft mice. o Figure S3: Mouse and human plasma stability of BCY-B5, BCY-C3, BCY-D1; o Figure S4: Pharmacokinetic profile of BCY-B2 in mouse; o Figure S5: In vivo metabolism of BCY-B2 in mice; o Figure S6: Confocal microscopy on HT1080 cells and BCY-C2 o Figure S7: PET imaging. Time-resolved whole-body maximum intensity projections of 68Ga-BCY-C2 in HT1080 xenograft mouse. €¢ Supplementary Methods: o Protein expression, o Phage selection, o Peptide synthesis, o Affinity determination by fluorescence polarization and SPR, o Radiolabeling, o Internalization, o Confocal microscopy, o Plasma protein binding, o Plasma stability of radiolabeled compounds, o µPET imaging, o Autoradiography, o Immunohistochemistry, o 6 references related to these methods.

Supplementary Data from BT8009; A Nectin-4 Targeting Bicycle Toxin Conjugate for Treatment of Solid Tumors

Supplementary Figure from Comprehensive Surfaceome Profiling to Identify and Validate Novel Cell-Surface Targets in Osteosarcoma

Supplementary Chemical Structures provides an inventory and structural diagrams of all Bicycle conjugates used in paper

Supplementary data are provided to further understand the pharmacokinetics and efficacy of BT5528 and EphA2 expression in tumors. Figure S1: Plasma concentration-time curves of BT5528 and MMAE following IV dosing of BT5528 1 in mouse, rat, and cynomolgus Monkey at 1 mg/kg (n=3 per species). Related to in vivo PK. Figure S2: EphA2 expression and tumour growth inhibition in CDX and PDX xenograft models are correlated. BT5528 3 mg/kg qw dosing has a range of anti-tumour activities across different cell-line derived and patient-derived xenograft models (*p<0.05, **p<0.01, ***p<0.001 2way ANOVA from D0 to last day of vehicle tumour measurement).Data used to generate Figure 3. Figure S3: EphA2 Immunohistochemistry staining of CDX and PDX xenograft models. A range of EphA2 expression is observed in stained tumour tissue from mouse xenograft models. Related to figure 3. Table T1: Ki values for BT5528, 1C1-mcMMAF and control compounds binding to human, mouse and rat Epha2 receptor. Ki values for Bicycle, BTC and DOTA analogues used to generate data in Figures 1, 4 and 5. Table T2: Binding affinities for BT5528 binding to EphA and EphB tyrosine kinases receptors determined by Surface Plasmon Resonance. Data defining BTC and Bicycle KD values for EphA2 orthologues. Table T3: In vitro ADME properties of BT5528 in mouse, rat, NHP and human: plasma protein binding, plasma stability and metabolic stability in hepatocytes. Related to in vitro ADME properties of BT5528. Table T4: Summary of experimentally determined and calculated PK Parameters for BT5528 and MMAE Following IV Dosing of 1 mg/kg BT5528 in Mouse, Rat and Cynomolgus Monkey. Related to in vivo PK Table T5a: Anti-Human EphA2 Antibody Binding Sites on CDX Cell Lines. Expression data used to generate Figure 3. Table T5b:Anti-Human EphA2 Antibody Binding Sites on PDX Cell Lines. Expression data used to generate Figure 3. Table T6: Doses of BT5528 and corresponding toxin (MMAE) used in preclinical toxicology studies compared with the clinical dose of MED-547 and corresponding toxin (MMAF)1. Related to the discussion (ADC versus BTC toxicology responses).

Supplementary information detailing methods used to generate in vitro and in vivo data presented in paper