Fluorescent BAPAD Dendrimeric Antigens Are Efficiently Internalized by Human Dendritic Cells

A new fluorescent dendrimeric antigen (DeAn) based on a dendron with amoxicilloyl terminal groups was synthesized. The synthesis was carried out using a novel class of all-aliphatic polyamide dendrimer (BisAminoalkylPolyAmide Dendrimers, or BAPAD) involving the direct condensation of 3,3′-diazidopivalic acid as a building block. Iterative azide reduction/amide formation increases the dendrimer generation. The BAPAD dendrimer was designed with a cystamine core. Reduction of the disulfide bond allows the incorporation of BAPAD dendrons into a 1,8-naphthalimide functionalized with a maleimide group. The fluorescence properties of DeAn were studied in PBS and compared with the properties of an equivalent dendron possessing amino-terminal groups. Both molecules shown high fluorescence quantum yields in PBS and could readily be visualized by fluorescence microscopy. DeAn was used as a synthetic antigen in a biomedical assay that tests their potential as an amoxicillin carrier in drug internalization by dendritic cells (DC) from tolerant and allergic patients. Cytometry data suggest that the dendrons are non-toxic and easily internalized by DCs, while confocal microscopy images indicate that the compounds are preferentially accumulated in the cytoplasm. These results indicate that BAPAD dendrons are good candidates for synthetic scaffolds for biomedical applications.

[1]  E. Baumann.  Ueber eine einfache Methode der Darstellung von Benzoësäureäthern , 1886 .

[2]  E. Pérez-Inestrosa,et al.  Immediate allergic reactions to cephalosporins: evaluation of cross-reactivity with a panel of penicillins and cephalosporins. , 2006, The Journal of allergy and clinical immunology.

[3]  C. Ornelas.  Brief Timelapse on Dendrimer Chemistry: Advances, Limitations, and Expectations , 2016 .

[4]  P. MacAry,et al.  Practical synthesis of maleimides and coumarin-linked probes for protein and antibody labelling via reduction of native disulfides. , 2009, Organic & biomolecular chemistry.

[5]  Vaibhav Jain,et al.  Dendrimer building toolkit: Model building and characterization of various dendrimer architectures , 2012, J. Comput. Chem..

[6]  Mohammad Ramezani,et al.  Gene delivery efficiency and cytotoxicity of heterocyclic amine-modified PAMAM and PPI dendrimers. , 2016, Materials science & engineering. C, Materials for biological applications.

[7]  D. Tomalia,et al.  Structure control within poly(amidoamine) dendrimers: size, shape and regio-chemical mimicry of globular proteins , 2003 .

[8]  Anne-Marie Caminade,et al.  Coordination chemistry with phosphorus dendrimers. Applications as catalysts, for materials, and in biology , 2016 .

[9]  H. Staudinger,et al.  Über neue organische Phosphorverbindungen III. Phosphinmethylenderivate und Phosphinimine , 1919 .

[10]  Yen Wei,et al.  A poly(amidoamine) dendrimer-based nanocarrier conjugated with Angiopep-2 for dual-targeting function in treating glioma cells , 2016 .

[11]  M. Blanca,et al.  Potential involvement of dendritic cells in delayed-type hypersensitivity reactions to beta-lactams. , 2006, The Journal of allergy and clinical immunology.

[12]  M. Schoenfisch,et al.  Anti-biofilm action of nitric oxide-releasing alkyl-modified poly(amidoamine) dendrimers against Streptococcus mutans. , 2016, Acta biomaterialia.

[13]  Stephen Z. D. Cheng,et al.  Tuning “thiol-ene” reactions toward controlled symmetry breaking in polyhedral oligomeric silsesquioxanes , 2014 .

[14]  E. Pérez-Inestrosa,et al.  Synthetic approach to gain insight into antigenic determinants of cephalosporins: in vitro studies of chemical structure-IgE molecular recognition relationships. , 2011, Chemical research in toxicology.

[15]  E. Pérez-Inestrosa,et al.  Large-scale dendrimer-based uneven nanopatterns for the study of local arginine-glycine-aspartic acid (RGD) density effects on cell adhesion , 2014, Nano Research.

[16]  Sanjeev Banerjee,et al.  PAMAM dendrimers as promising nanocarriers for RNAi therapeutics , 2015 .

[17]  C. Schotten.  Ueber die Oxydation des Piperidins , 1883 .

[18]  E. Pérez-Inestrosa,et al.  Dendrimeric antigen-silica particle composites: an innovative approach for IgE quantification. , 2013, Journal of materials chemistry. B.

[19]  S. Saha,et al.  Influence of the Structure of the Amino Group and Polarity of the Medium on the Photophysical Behavior of 4-Amino-1,8-naphthalimide Derivatives , 2002 .

[20]  G. Ciccotti,et al.  Numerical Integration of the Cartesian Equations of Motion of a System with Constraints: Molecular Dynamics of n-Alkanes , 1977 .

[21]  K Schulten,et al.  VMD: visual molecular dynamics. , 1996, Journal of molecular graphics.

[22]  M. Pirmohamed,et al.  Immunological Principles of Adverse Drug Reactions , 2000, Drug safety.

[23]  E. Bahadır,et al.  Poly(amidoamine) (PAMAM): An emerging material for electrochemical bio(sensing) applications. , 2016, Talanta.

[24]  George M Whitesides,et al.  Polyvalent Interactions in Biological Systems: Implications for Design and Use of Multivalent Ligands and Inhibitors. , 1998, Angewandte Chemie.

[25]  Anna M. Maj,et al.  Efficient Catalytic Hydrogenation of N‐Unsubstituted Cyclic Imides to Cyclic Amines , 2014 .

[26]  W. Hung,et al.  Maleimide-Functionalized PEI600 Grafted Polyurethane: Synthesis, Nano-Complex Formation with DNA and Thiol-Conjugation of the Complexes for Dual DNA Transfection , 2015 .

[27]  M. Bachmann,et al.  The Bidirectional Crosstalk between Human Dendritic Cells and Natural Killer Cells , 2011, Journal of Innate Immunity.

[28]  Yi Zhang,et al.  Effect of poly(amidoamine) dendrimers on the structure and activity of immune molecules. , 2015, Biochimica et biophysica acta.

[29]  E. Pérez-Inestrosa,et al.  Synthesis of all-aliphatic polyamide dendrimers based on a 3,3′-diaminopivalic acid scaffold , 2015 .

[30]  Junmei Wang,et al.  Development and testing of a general amber force field , 2004, J. Comput. Chem..

[31]  S. Pilard.  A stereospecific synthesis of () a-conhydrine and () -conhydrine. , 1984 .

[32]  P. Bosch,et al.  The synthesis of a novel 1,8-naphthalimide based PAMAM-type dendron and its potential for light-harvesting , 2009 .

[33]  Laura M Lechuga,et al.  Highly sensitive dendrimer-based nanoplasmonic biosensor for drug allergy diagnosis. , 2015, Biosensors & bioelectronics.

[34]  S. Michnick,et al.  Convergent preparation and photophysical characterization of dimaleimide dansyl fluorogens: elucidation of the maleimide fluorescence quenching mechanism. , 2007, Journal of the American Chemical Society.

[35]  Albert M. Brouwer,et al.  Standards for photoluminescence quantum yield measurements in solution (IUPAC Technical Report) , 2011 .

[36]  Mark E. B. Smith,et al.  Homogeneous Bispecifics by Disulfide Bridging , 2014, Bioconjugate chemistry.

[37]  NMR Studies and Molecular Dynamic Simulation of Synthetic Dendritic Antigens , 2011 .

[38]  Yuan Ping,et al.  Structure‐Invertible Nanoparticles for Triggered Co‐Delivery of Nucleic Acids and Hydrophobic Drugs for Combination Cancer Therapy , 2015 .