Development of 5D3-DM1: A Novel Anti-Prostate-Specific Membrane Antigen Antibody-Drug Conjugate for PSMA-Positive Prostate Cancer Therapy

Huang, Colin T.; Guo, Xin; Bařinka, Cyril; Lupold, Shawn E.; Pomper, Martin G.; Gabrielson, Kathleen L.; Raman, Venu; Artemov, Dmitri; Hapuarachchige, Sudath

doi:10.1021/acs.molpharmaceut.0c00457

Cited by 23 publications

(16 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With this protocol, we were able to monitor and quantify the entire PSMA/probe complex formation and endocytosis process in living cells in real-time. By quantifying the effects of different types of endocytosis inhibitors on PSMA-mediated endocytosis, we confirmed that the clathrin-dependent endocytosis pathway plays the most important role in the endocytosis of PSMA 53 , 54 , while the contribution of the caveolin-dependent and micropinocytosis pathway is quite limited. That is consistent with the established results that most transmembrane receptors are internalized via a clathrin-dependent mechanism, as recently shown by Matthias et al, who did so using stimulated emission depletion (STED) nanoscopy 55 .…”

Section: Discussionmentioning

confidence: 64%

A prostate-specific membrane antigen activated molecular rotor for real-time fluorescence imaging

et al. 2021

Self Cite

View full text Add to dashboard Cite

Surgery is an efficient way to treat localized prostate cancer (PCa), however, it is challenging to demarcate rapidly and accurately the tumor boundary intraoperatively, as existing tumor detection methods are seldom performed in real-time. To overcome those limitations, we develop a fluorescent molecular rotor that specifically targets the prostate-specific membrane antigen (PSMA), an established marker for PCa. The probes have picomolar affinity (IC50 = 63-118 pM) for PSMA and generate virtually instantaneous onset of robust fluorescent signal proportional to the concentration of the PSMA-probe complex. In vitro and ex vivo experiments using PCa cell lines and clinical samples, respectively, indicate the utility of the probe for biomedical applications, including real-time monitoring of endocytosis and tumor staging. Experiments performed in a PCa xenograft model reveal suitability of the probe for imaging applications in vivo.

show abstract

Section: Discussionmentioning

confidence: 64%

A prostate-specific membrane antigen activated molecular rotor for real-time fluorescence imaging

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…Similarly to other PSMA-specific antibody fragments [72,76], we observed signal originated from 5D3 conjugates in kidneys and liver that is connected with metabolism and clearance of recombinant proteins. Described distribution should be carefully followed particularly in future development of radionuclide or cytotoxic drug 5D3 conjugates because of potential risk of kidney toxicity [45]. The behavior of original 5D3 mAb in long time lapse was also similar to other high-affinity mouse antibodies [77], as the mAb revealed the peak of accumulation at days 4-6 p.i.…”

Section: Discussionmentioning

confidence: 99%

“…In this report, we exploit the superior features of our newly developed 5D3 mAb, including sub-nanomolar affinity and a high specificity for native PSMA [43]. These characteristics make the 5D3 mAb particularly suitable for in vivo applications [44][45][46]. Here, we cloned, heterologously expressed, purified, and characterized engineered scFv and Fab fragments of 5D3.…”

Section: Introductionmentioning

confidence: 99%

Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization

Nováková

Belousova

Foss

et al. 2020

IJMS

Self Cite

View full text Add to dashboard Cite

Prostate-Specific Membrane Antigen (PSMA) is an established biomarker for the imaging and experimental therapy of prostate cancer (PCa), as it is strongly upregulated in high-grade primary, androgen-independent, and metastatic lesions. Here, we report on the development and functional characterization of recombinant single-chain Fv (scFv) and Fab fragments derived from the 5D3 PSMA-specific monoclonal antibody (mAb). These fragments were engineered, heterologously expressed in insect S2 cells, and purified to homogeneity with yields up to 20 mg/L. In vitro assays including ELISA, immunofluorescence and flow cytometry, revealed that the fragments retain the nanomolar affinity and single target specificity of the parent 5D3 antibody. Importantly, using a murine xenograft model of PCa, we verified the suitability of fluorescently labeled fragments for in vivo imaging of PSMA-positive tumors and compared their pharmacokinetics and tissue distribution to the parent mAb. Collectively, our data provide an experimental basis for the further development of 5D3 recombinant fragments for future clinical use.

show abstract

“…This limitation was partially overcome by the development of another anti-PSMA antibody, J591, that binds extracellular epitopes of PSMA, and was largely used in preclinical and clinical trials (ClinicalTrials.gov Identifier: NCT00195039; NCT02410577). Other mAbs directed to PSMA extracellular epitopes, as well as their conjugates, are being evaluated in a variety of experimental and preclinical models [ 23 , 24 , 25 ], but only IgGD2B [ 26 ], similarly to J591 [ 27 ], has been extensively characterized, demonstrating low tumor-to-blood ratios that can occur owing to relatively slow clearance of intact antibodies compared with fragments [ 28 ]. Different groups analyzed IgGD2B tumor-targeting capacity ( Table 1 , refs.…”

Section: Psma-targeted Diagnosis/therapy With Monoclonal Antibodiesmentioning

confidence: 99%

Validity of Anti-PSMA ScFvD2B as a Theranostic Tool: A Narrative-Focused Review

et al. 2021

View full text Add to dashboard Cite

Prostate cancer (PCa) is the second leading cause of cancer among men, and its diagnosis and adequate staging are fundamental. Among the biomarkers identified in recent years for PCa management, prostate-specific-membrane-antigen (PSMA), physiologically expressed at a low level on healthy prostate and in other normal tissues and highly overexpressed in PCa, represents a reliable marker ideal for imaging and therapy. The development of anti-PSMA antibodies, such as D2B, demonstrated slow clearance of intact antibodies compared with fragments resulting in low tumor-to-blood ratios; however, the modular structural and functional nature of antibodies allowed the generation of smaller fragments, such as scFvs. In this review of the anti-PSMA antibody fragment scFvD2B, we combined further characterization of its biomolecular and tissue cross-reactivity characteristics with a comprehensive summary of what has already been performed in preclinical models to evaluate imaging and therapeutic activities. A molecular dynamics study was performed, and ScFvD2B occupied a limited conformational space, characterized by low-energy conformational basins, confirming the high stability of the protein structure. In the cross-reactivity study, the weak/absent immunoreactivity in non-tumor tissues was comparable to the PSMA expression reported in the literature. Biodistribution studies and therapeutic treatments were conducted in different animal models obtained by subcutaneous or locoregional injection of PSMA-positive-versus-negative xenografts. The maximum tumor uptake was observed for 123I(SPECT), 124I(PET), and optical imaging, which avoids kidney accumulation (compared with radiometals) and leads to an optimal tumor-to-kidney and tumor-to-background ratios. Regarding its possible use in therapy, experimental data suggested a strong and specific antitumor activity, in vitro and in vivo, obtained using CAR-T or NK-92/CAR cells expressing scFvD2B. Based on presented/reviewed data, we consider that scFvD2B, due to its versatility and robustness, seems to: (i) overcome some problems observed in other studied scFvs, very often relatively unstable and prone to form aggregates; (ii) have sufficient tumor-to-background ratios for targeting and imaging PSMA-expressing cancer; (iii) significantly redirect immune killing cells to PSMA-positive tumors when inserted in second-generation CAR-T or NK-92/CAR cells. These data suggest that our product can be considered the right reagent to fill the gap that still exists in PCa diagnosis and treatment.

show abstract

Development of 5D3-DM1: A Novel Anti-Prostate-Specific Membrane Antigen Antibody-Drug Conjugate for PSMA-Positive Prostate Cancer Therapy

Cited by 23 publications

References 42 publications

A prostate-specific membrane antigen activated molecular rotor for real-time fluorescence imaging

A prostate-specific membrane antigen activated molecular rotor for real-time fluorescence imaging

Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization

Validity of Anti-PSMA ScFvD2B as a Theranostic Tool: A Narrative-Focused Review

Contact Info

Product

Resources

About