A Wireframe DNA Cube: Antibody Conjugate for Targeted Delivery of Multiple Copies of Monomethyl Auristatin E

Albumin-nucleic acid biomolecular drug designs offer modular multifunctionalization and extended circulatory halflife. However, stability issues associated with conventional DNA nucleotides and maleimide bioconjugation chemistries limit the clinical potential. This work aims to improve the stability of this thiol conjugation and nucleic acid assembly by employing a fasthydrolyzing monobromomaleimide (MBM) linker and nucleaseresistant nucleotide analogues, respectively. The biomolecular constructs were formed by site-selective conjugation of a 12-mer oligonucleotide to cysteine 34 (Cys34) of recombinant human albumin (rHA), followed by annealing of functionalized complementary strands bearing either a fluorophore or the cytotoxic drug monomethyl auristatin E (MMAE). Formation of conjugates and assemblies was confirmed by gel shift analysis and mass spectrometry, followed by investigation of serum stability, neonatal Fc receptor (FcRn)-mediated cellular recycling, and cancer cell killing. The MBM linker afforded rapid conjugation to rHA and remained stable during hydrolysis. The albumin-nucleic acid biomolecular assembly composed of stabilized oligonucleotides exhibited high serum stability and retained FcRn engagement mediating FcRn-mediated cellular recycling. The MMAE-containing assembly exhibited cytotoxicity in the human MIA PaCa-2 pancreatic cancer cell line with an IC50 of 342 nM, triggered by drug release from breakdown of an acid-labile linker. In summary, this work presents rHA-nucleic acid module-based assemblies with improved stability and retained module functionality that further promotes the drug delivery potential of this biomolecular platform.

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“…LC-MS Confirmation of Conjugation. LC-MS was used for confirmation of functionalized oligonucleotide masses as described in ref 50.…”

Section: ■ Discussionmentioning

confidence: 99%

Albumin Biomolecular Drug Designs Stabilized through Improved Thiol Conjugation and a Modular Locked Nucleic Acid Functionalized Assembly

et al. 2022

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“…To synthesize the DNA-antibody conjugate we used a so-called LDLR reagent for site-directed labeling reaction that preferentially targets the hinge region of the antibody to insert an azide handle (Figure 2a). [37,38] The azidelabeled antibody was then submitted to further conjugation with a dibenzocyclooctyne (DBCO)-modified DNA sequence. A strain-promoted azide-alkyne cycloaddition (SPAAC) reaction yielded the DNA-antibody construct (Figure 2b and Figure S2, Supporting Information).…”

Section: Site-directed Labeling Of Antibodiesmentioning

confidence: 99%

Protein‐Templated Reactions Using DNA‐Antibody Conjugates

Pellejero

Nijenhuis

Ricci

et al. 2022

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DNA-templated chemical reactions have found wide applications in drug discovery, programmed multistep synthesis, nucleic acid detection and targeted drug delivery. The control of these reactions has, however, been limited to nucleic acid hybridization as a means to direct the proximity between reactants.In this work we introduce a system capable of translating protein-protein binding events into a DNAtemplated reaction which leads to the covalent formation of a product. We achieve protein-templated reactions by employing two DNA-antibody conjugates that are both able to recognize the same target protein and to colocalize a pair of reactant DNA strands able to undergo a click reaction. We engineered two individual systems each responsive to human serum albumin (HSA) and human IgG and we demonstrated that, while no reaction occurs in the absence of proteins, both protein-templated reactions can occur simultaneously in the same solution without any inter-system crosstalk.

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“…Finally, Gothelf et al have designed an interesting ADC based on the use of a DNA nanostructure that would allow for high loading of the drug and exquisite control of DAR, resulting in homogeneous and well-defined ADCs, key features required for their clinical efficiency and safety [ 126 ]. The programmability of oligonucleotides should allow for the precise design of DNA-based frameworks with suitable modifications for a controlled and precise arrangement of the drug in a predefined ratio.…”

Section: Chemistry and Biology Of Marine Antibody-drug Conjugatesmentioning

confidence: 99%

Antibody-Drug Conjugates Containing Payloads from Marine Origin

et al. 2022

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Antibody-drug conjugates (ADCs) are an important class of therapeutics for the treatment of cancer. Structurally, an ADC comprises an antibody, which serves as the delivery system, a payload drug that is a potent cytotoxin that kills cancer cells, and a chemical linker that connects the payload with the antibody. Unlike conventional chemotherapy methods, an ADC couples the selective targeting and pharmacokinetic characteristics related to the antibody with the potent cytotoxicity of the payload. This results in high specificity and potency by reducing off-target toxicities in patients by limiting the exposure of healthy tissues to the cytotoxic drug. As a consequence of these outstanding features, significant research efforts have been devoted to the design, synthesis, and development of ADCs, and several ADCs have been approved for clinical use. The ADC field not only relies upon biology and biochemistry (antibody) but also upon organic chemistry (linker and payload). In the latter, total synthesis of natural and designed cytotoxic compounds, together with the development of novel synthetic strategies, have been key aspects of the consecution of clinical ADCs. In the case of payloads from marine origin, impressive structural architectures and biological properties are observed, thus making them prime targets for chemical synthesis and the development of ADCs. In this review, we explore the molecular and biological diversity of ADCs, with particular emphasis on those containing marine cytotoxic drugs as the payload.

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A Wireframe DNA Cube: Antibody Conjugate for Targeted Delivery of Multiple Copies of Monomethyl Auristatin E

Cited by 27 publications

References 46 publications

Albumin Biomolecular Drug Designs Stabilized through Improved Thiol Conjugation and a Modular Locked Nucleic Acid Functionalized Assembly

Albumin Biomolecular Drug Designs Stabilized through Improved Thiol Conjugation and a Modular Locked Nucleic Acid Functionalized Assembly

Protein‐Templated Reactions Using DNA‐Antibody Conjugates

Antibody-Drug Conjugates Containing Payloads from Marine Origin

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