Radionuclide therapy using ABD-fused ADAPT scaffold protein: Proof of Principle

Garousi, Javad; Witting, Emma von; Borin, Jesper; Vorobyeva, Anzhelika; Altai, Mohamed; Vorontsova, Olga; Konijnenberg, Mark; Oroujeni, Maryam; Orlova, Anna; Tolmachev, Vladimir; Hober, Sophia

doi:10.1016/j.biomaterials.2020.120381

Cited by 16 publications

(16 citation statements)

References 44 publications

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“…where it reached a plateau of 5% ID/g. This was lower compared to the homologous ADAPT6-ABD labeled at the C-terminus with 177 Lu, 177 Lu-DOTA-ABD 035 -ADAPT6, which has been evaluated in a similar mouse model with SKOV-3 tumor xenografts [28]. In that study, the tumor uptake increased up to 26 ± 4% ID/g at 24 h. The higher tumor uptake of 177 Lu-labeled construct is likely a consequence of its higher blood radioactivity, 17 ± 2% IA/g at 24 h, and thus higher bioavailability compared to 99m Tc-ADAPT6-ABD-mcDM1, with a blood radioactivity of 5.0% ID/g at 24 h. At 24 h, the uptake in liver of 177 Lu-DOTA-ABD 035 -ADAPT6 (4.8 ± 0.3% ID/g) and technetium-99 labeled ADAPT6-ABD-mcDM1 (4.83 ± 0.27% ID/g) were similar, but the uptake in the kidneys was significantly lower for 177 Lu-DOTA-ABD 035 -ADAPT6 (10.9 ± 0.7% ID/g) than for ADAPT6-ABD-mcDM1 (83 ± 4% ID/g).…”

Section: Discussionmentioning

confidence: 95%

“…The intracellular fraction was released by sonication. The proteins were purified by affinity chromatography on a column with immobilized human serum albumin (HSA) as previously described [28]. Briefly, the cell lysates were loaded on the column after equilibration with 1xTST buffer (25 mM Tris-HCl, 1 mM EDTA, 0.2 M NaCl, 0.05% Tween 20, pH 8.0), followed by washing with 1xTST and 5 mM NH 4 Ac (pH 5.5).…”

Section: Production and Purification Of Adapt Fusion Proteinsmentioning

confidence: 99%

“…Previously, we have found that it is possible to extend the plasma half-life of ESPs by fusion to an albumin binding domain (ABD), which consequently should improve its bioavailability [25][26][27][28]. Half-life extension occurs by binding of the ABD to serum albumin in the blood, leading to an increase in the overall size of the complex that exceeds the filtration cut-off of the kidneys (approximately 60 kDa).…”

Section: Introductionmentioning

confidence: 99%

“…A particularly useful ABD variant is ABD 035 with femtomolar binding affinity (K D ) to human serum albumin (HSA) [26]. Moreover, we have demonstrated that an ABD-fused ADAPT6 labeled with 177 Lu can be used to efficiently target human cancer xenografts with high HER2 expression in mice [28]. It was also found that the relative position of ABD and ADAPT6 influences the biodistribution, and that placement of the ABD at the C-terminus of ADAPT6 was particularly favorable [28].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, we have demonstrated that an ABD-fused ADAPT6 labeled with 177 Lu can be used to efficiently target human cancer xenografts with high HER2 expression in mice [28]. It was also found that the relative position of ABD and ADAPT6 influences the biodistribution, and that placement of the ABD at the C-terminus of ADAPT6 was particularly favorable [28]. We have therefore used an analogous architecture for the drug conjugates in this study, with ADAPT6 followed by the ABD.…”

Section: Introductionmentioning

confidence: 99%

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Targeting HER2 Expressing Tumors with a Potent Drug Conjugate Based on an Albumin Binding Domain-Derived Affinity Protein

et al. 2021

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Albumin binding domain derived affinity proteins (ADAPTs) are a class of small and folded engineered scaffold proteins that holds great promise for targeting cancer tumors. Here, we have extended the in vivo half-life of an ADAPT, targeting the human epidermal growth factor receptor 2 (HER2) by fusion with an albumin binding domain (ABD), and armed it with the highly cytotoxic payload mertansine (DM1) for an investigation of its properties in vitro and in vivo. The resulting drug conjugate, ADAPT6-ABD-mcDM1, retained binding to its intended targets, namely HER2 and serum albumins. Further, it was able to specifically bind to cells with high HER2 expression, get internalized, and showed potent toxicity, with IC50 values ranging from 5 to 80 nM. Conversely, no toxic effect was found for cells with low HER2 expression. In vivo, ADAPT6-ABD-mcDM1, radiolabeled with 99mTc, was characterized by low uptake in most normal organs, and the main excretion route was shown to be through the kidneys. The tumor uptake was 5.5% ID/g after 24 h, which was higher than the uptake in all normal organs at this time point except for the kidneys. The uptake in the tumors was blockable by pre-injection of an excess of the monoclonal antibody trastuzumab (having an overlapping epitope on the HER2 receptor). In conclusion, half-life extended drug conjugates based on the ADAPT platform of affinity proteins holds promise for further development towards targeted cancer therapy.

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Section: Discussionmentioning

confidence: 95%

Section: Production and Purification Of Adapt Fusion Proteinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Targeting HER2 Expressing Tumors with a Potent Drug Conjugate Based on an Albumin Binding Domain-Derived Affinity Protein

et al. 2021

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GPC3-targeted immunoPET imaging of hepatocellular carcinomas

Zhang

et al. 2022

Eur J Nucl Med Mol Imaging

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Evaluation of a novel 177Lu-labelled therapeutic Affibody molecule with a deimmunized ABD domain and improved biodistribution profile

Liu,

Oroujeni,

Liao

et al. 2024

Eur J Nucl Med Mol Imaging

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Purpose Fusion of Affibody molecules with an albumin-binding domain (ABD) provides targeting agents, which are suitable for radionuclide therapy. To facilitate clinical translation, the low immunogenic potential of such constructs with targeting properties conserved is required. Methods The HER2-targeting Affibody molecule ZHER2:2891 was fused with a deimmunized ABD variant and DOTA was conjugated to a unique C-terminal cysteine. The novel construct, PEP49989, was labelled with 177Lu. Affinity, specificity, and in vivo targeting properties of [177Lu]Lu-PEP49989 were characterised. Experimental therapy in mice with human HER2-expressing xenografts was evaluated. Results The maximum molar activity of 52 GBq/µmol [177Lu]Lu-PEP49989 was obtained. [177Lu]Lu-PEP49989 bound specifically to HER2-expressing cells in vitro and in vivo. The HER2 binding affinity of [177Lu]Lu-PEP49989 was similar to the affinity of [177Lu]Lu-ABY-027 containing the parental ABD035 variant. The renal uptake of [177Lu]Lu-PEP49989 was 1.4-fold higher, but hepatic and splenic uptake was 1.7-2-fold lower than the uptake of [177Lu]Lu-ABY-027. The median survival of xenograft-bearing mice treated with 21 MBq [177Lu]Lu-PEP49989 (> 90 days) was significantly longer than the survival of mice treated with vehicle (38 days) or trastuzumab (45 days). Treatment using a combination of [177Lu]Lu-PEP49989 and trastuzumab increased the number of complete tumour remissions. The renal and hepatic toxicity was minimal to mild. Conclusion In preclinical studies, [177Lu]Lu-PEP49989 demonstrated favourable biodistribution and a strong antitumour effect, which was further enhanced by co-treatment with trastuzumab.

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Radionuclide therapy using ABD-fused ADAPT scaffold protein: Proof of Principle

Cited by 16 publications

References 44 publications

Targeting HER2 Expressing Tumors with a Potent Drug Conjugate Based on an Albumin Binding Domain-Derived Affinity Protein

Targeting HER2 Expressing Tumors with a Potent Drug Conjugate Based on an Albumin Binding Domain-Derived Affinity Protein

GPC3-targeted immunoPET imaging of hepatocellular carcinomas

Evaluation of a novel 177Lu-labelled therapeutic Affibody molecule with a deimmunized ABD domain and improved biodistribution profile

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