BCCHES
`
`
`
`A PUBLICATION OF THE AMERICAN CHEMICAL SOCIETY
`
`Foo 26 208
`\
`
`/
`
`Thr(OH)-Cys-Thr—Lys
`
`February 2008
`Volume 19, Number 2
`http://pubs.acs.org/BC
`
`4-D-Phe-Cys-TyrD-7rp
`.
`55
`
`
`
`
`DOTA-Peptide Conjugates (see p. 397)
`
`Evergeen Ex. 1014
`1 of 20
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`Evergeen Ex. 1014
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`

`Bioconjugate
`ChemIStry
`
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`© Copyright 2008 American Chemical Society
`
`Evergeen Ex. 1014
`2 of 20
`
`

`

`Bioconjugate
`Chemistry
`
`EDITOR-IN-CHIEF
`CLAUDE F. MEARES
`Department of Chemistry, University of Califomia, One Shields Avenue, Da vis, Califomia 95616-
`(530) 752-3360; Fax (530) 752-8938; E-mail cfmeares@ucdavis.edu
`SENIOR EDITORS
`Paul S. Miller, The Johns Hopkins University, Baltimore, Maryland
`Pe1er Senler, Seanle Genetics, hie., Bothell, Washington
`ASSOCIATE EDITORS
`Ferenc Hudecz. Eotvos lorw1d University. Budapest, Hungary
`Ma ayuki Yokoyama. Kanagawa Academy of Science and Technology. Kawa aki, Japan
`
`Carolyn J. Anderson
`The Edward Mallinckrodt lnstitwe
`of Radiology
`Jacqueline Barton
`California Institute of Technology
`Jean-Paul Behr
`lAboratoire de Chimie Genetique
`Darryl Bomhop
`Vanderbilt University
`Cynlhia J. Burrows
`University of Utah
`Chien-H ing Ken Chang
`lmmunomedics
`Xiaoyuan Chen
`Stanford University School of Medicine
`Patrick Couvreur
`Universiti de Paris-Sud
`Sylvia Daunen
`University of Ke11111cky
`Mark E. Davis
`California lnstilllte of Technology
`Pe1er B. Dervan
`California Institute of Technology
`Gene M. Dubowchik
`Bristol-Myers Squibb
`Rulh Duncan
`University of Cardiff
`David Filpula
`Enwn Phannace111ica/s
`M. G. Finn
`Tile Scripps Research lnsriwte
`Shiroh Fulak:i
`Kyoto University
`David A. Goodwin
`Stanford University (retired)
`Anna Grandas
`UniversitaJ de Barcelona
`Anoe Gruaz-Guyon
`Faculte de Medecine Saint Antoine
`Philip R. Hamano
`Wyeth Research
`M. Frederick Hawlhome
`International Institute of Na110 and
`Molecular Medicine
`John E. Hean;!
`University of California, Berkeley
`ed D. Heindel
`Lehigh University
`Wim E. Hennink
`Utrecht University
`Greg Hermanson
`Pierce Chemical Company
`Don J. Hnatowich
`University of Massachusens Medical Center
`Allan S. Hoffman
`University of Washington
`Kenneth A. Jacobson
`National Institutes of Health
`
`ADVISORY BOARD
`AJe~ander V. Kabanov
`U11iversity of Nebraska Medical Cemer
`Kazunori Kataoka
`The University of Tokyo
`John A. Ka1zenellenbogen
`University of Illinois
`Sung Wan Kim
`University of Utah
`Thomas Kissel
`Phillipps•University of Marb11rg
`Kenj i Kono
`Osaka Prefecwre University
`Jindrich Kopecek
`University of Utah
`Robert Langer
`Massac/111se11s lnstitwe of Tech110/ogy
`Chris Leamon
`Endocyte P/,am,aceuticals, Inc.
`Michael R. Lewis
`University of Missouri-Columbia
`Shuang Liu
`Purdue University
`Harri Ulnnberg
`University of Turku
`H. R. Maecke
`University Hospital Basel
`Hiro bi Maeda
`Sojo University
`Thomas J. Meade
`Northwestern University
`Oleg Melnyk
`Biological lnstiture of Lille
`Sherie L. Morrison
`University of California, Los Angeles
`Pe1er E. Nielsen
`University of Copenhagen
`Jong-Sang Park
`Seoul National University
`Tae Gwan Park
`Korea Adl/Qnced lnstiture of Science and Tech110/ogy
`Glenn D. Pres1wich
`University of Utah
`Tariq Rana
`University of Massachusetrs Medical School
`Steve Regen
`Lehigh University
`Ralph Reisfeld
`Scripps Clinic and Research Foundation
`Annie C. Roche
`Universite de'Orleans
`Sieve Roffler
`Academia Sinica
`Keith Rose
`Ge11eProt Inc.
`
`P. A. Schubiger
`Par,/ Scherrer fost illlt
`Peter G. Schultz
`The Scripps Research lnsti1111e
`John E. Shively
`Beckman Research lnstit111e
`Caroline Springer
`Institute of Cancer Research, CRC Labs
`Suresh Srivastava
`Brookhaven National Laboratory
`Francis Szoka
`University of California, San Francisco
`David H. Thompson
`P11rd11e University
`David A. Tirrell
`California /nstiwre of Technology
`Donald A. Tomalia
`Dendriric Na1wtecl111ologies Inc.
`Vladimir P. Torchilin
`Northeastern University
`Pamela A. Trail
`Bristol-Myers Squibb
`Eishun Tsuchida
`Waseda University
`M. Garca H. Vicen1e
`louisia11a Stale Uni versify
`Pierre Vierling
`Universite de Nice Sophia-Amipolis
`Wynn A. Volkert
`University of Missouri, Columbia
`Ernst Wagner
`Ludwig Maximilian University
`Ralph Weissleder
`Harl/Qrd Medical School
`Michael J. Welch
`Washington University School of Medicine
`Eric Wickstrom
`Thomas Jefferson University
`D. Scon Wilbur
`University of Washington
`Meir Wilchek
`Weiunann lnstiwte of Science
`Karen L. Wooley
`Washington University in St. louis
`Anna Wu
`University of California, Los A11geles
`John Yates
`The Scripps Research Institute
`Samuel Zalipsky
`lntradigm, Inc.
`Michael Zalutsky
`Duke University Medical Center
`
`Evergeen Ex. 1014
`3 of 20
`
`

`

`Volume 19, Number 2
`FEBRUARY 2008
`
`BCCHES 19(2) 391-580 (2008)
`ISSN 1043-1802
`Registered in U.S. Patefl/ and Trademark Office
`Copyright 2008 by the American Chemical Society
`
`Bioconjugate
`Chemistry®
`
`REVIEWS
`
`The Synthesis and Chelation Chemistry of OCTA- Peptide Conjugates
`
`391
`
`Luis M. De Le6n-Rodriguez and
`Zoltan Kovacs*
`
`COMMUNICATIONS
`
`A family of linear-dendritic "hybrid" polymers containing multiple
`modular functionalities are designed and synthesized using a
`three-step, aqueous approach. These polymers are functionalized
`with a peptide targeting ligand that specifically binds to glucose(cid:173)
`regulated protein-78 kDa (GRP-78), a clinically relevant tumor
`antigen identified in human cancer patients. The resultant systems
`can condense plasmid DNA into small nanoparticle structures and
`transfect cells expressing GRP-78 with efficiencies that exceed
`branched polyethylenimine (bPEI). As such, they may be useful in
`clinical cancer gene therapy applications.
`
`Kris C. Wood, Samira M. Azarin, Wadih Arap,
`Renata Pasqualini, Robert Langer,• and
`Paula T. Hammond'
`Bioconjugate Chem. 2008, 19, 403 (cid:127)
`Tumor-Targeted Gene Delivery Using
`Molecularly Engineered Hybrid Polymers
`Functionalized with a Tumor-Homing Peptide
`
`Evergeen Ex. 1014
`4 of 20
`
`

`

`Activation of the A2A receptor, a G protein-coupled receptor
`(G PCR) , for extracellular adenosine, is anti-aggregatory
`in
`platelets and anti-inflammatory. PAMAM dendrimers were
`covalently coupled to multiple copies of an A2A agonist, the
`nucleoside CGS21680, and characterized spectroscopically. A
`5
`inhibited
`fluorescent
`PAMAM-CGS21680
`conjugate
`aggregation of washed human platelets and was internalized.
`We envision that our multivalent dendrimer conjugates may
`to
`the
`improve overall pharmacological profiles compared
`monovalent GPCR ligands.
`
`Bioconjugate Chem., Vol. 19, No. 2, 2008 SA
`
`Yoonkyung Kim, Beatrice Hechler, Athena M. Klutz,
`Christian Gachet, and Kenneth A. Jacobson'
`Bioconjugate Chem. 2008, 19, 406 (cid:127)
`Toward Multivalent Signaling across G
`Protein-Coupled Receptors from
`Poly(amidoamine) Dendrimers
`
`Acllvallon of
`Plale/el AQfl~•llon
`
`byADP "'
`-,-G~b-, •
`_..,...
`D mmK JII
`Antl-Aggregatory Effect i
`
`-
`
`m~
`
`The detection of the human a-thrombin protein by MRI is
`reported by designing a thrombin targeted MRI contrast agent.
`The contrast agent
`is composed of
`thrombin aptamer
`functionalized superparamagnetic iron oxide nanoparticles. The
`detection of thrombin is based on the increase in the size of
`nanoparticle assembly, which leads to a change in brightness of
`the image. A detectable change in MR signal is observed with
`25 nM thrombin in human serum.
`
`Mehmet Veysel Yigit, Debapriya Mazumdar, and
`Yi Lu'
`Bioconjugate Chem. 2008, 19, 412
`MRI Detection of Thrombin with Aptamer
`Functionalized Superparamagnetic Iron Oxide
`Nanopartlcles
`
`Evergeen Ex. 1014
`5 of 20
`
`

`

`Bioconjugate Chem., Vol. 19, No. 2, 2008 7A
`
`A series of new lipophilic peptide vectors that m1m1c the
`recognition found in nucleic acid-protein interactions have been
`synthesized. These lipophilic peptides show minimal cytotoxicity
`and enhanced in vitro gene transfection activity.
`
`Carla A. H. Prata, Xiao-Xiang Zhang, Dan Luo,
`Thomas J. McIntosh, Philippe Barthelemy, and
`Mark W. Grinstatr
`Bioconjugate Chem. 2008, 19, 418 (cid:127)
`Lipophillc Peptides for Gene Delivery
`
`n = 4; n = 6; n = 8; n = 10
`
`ARTICLES
`
`Applications of Mesenchymal Stem Cells Labeled with Tat Peptide
`Conjugated Quantum Dots to Cell Tracking in Mouse Body
`
`421
`(cid:127)
`
`Yun Lei, Haiyang Tang, Lide Yao,
`Richeng Yu, Meifu Feng: and Bingsuo Zou•
`
`General Structure-Activity Relationship for Poly(glycoamidoamine)s: The
`Effect of Amine Density on Cytotoxicity and DNA Delivery Efficiency
`
`428 Chen-Chang Lee, Yemin Liu, and
`Theresa M. Reineke·
`
`Synthesis and Evaluation of 18F- and 11C-Labeled
`Phenyl-Galactopyranosides as Potential Probes for in Vivo Visualization
`of Lacz Gene Expression using Positron Emission Tomography
`
`441
`
`Sofie Gelen, Christophe Deroose,
`Tjibbe de Groot, Salish K. Chitneni,
`Rik Gijsbers, Zeger Debyser,
`Luc Mortelmans, Alfons Verbruggen, and
`Guy Bormans•
`
`Labeling of Fatty Acid Ligands with the Strong Electrophilic Metal
`Fragment [99'"Tc(N)(PNP)J + (PNP = Diphosphane Ligand)
`
`PEGylated Oendrimers with Core Functionality for Biological Applications
`
`Molecular Scale Architecture: Engineered Three- And Four-Way Junctions
`
`Enzyme-Directed Positioning of Nanoparticles on Large DNA Templates
`
`Semisynthetic Analogues of PSC-RANTES, a Potent Anti-HIV Protein
`
`DNA Binding, Cleavage, and Cytotoxic Activity of the Preorganized
`Dinuclear Zinc(II) Complex of Triazacyclononane Derivatives
`
`450 Emiliano Gazzola, Elisa Benini,
`Micol Pasquali, Peter Mirtschink,
`Martin Walther, Hans-Jurgen Pietzsch,
`Licia Uccelli, Alessandra Boschi,
`Cristina Bolzati, and Adriano Duatti•
`
`461 Steven J. Guillaudeu, Megan E. Fox,
`Yarah M. Haidar, Edward E. Dy,
`Francis C. Szoka, and Jean M. J. Frechet*
`
`470 Stephanie Wilkinson, Michael Diechtierow,
`(cid:127)
`A. August Estabrook, Falk Schmidt,
`Michael HOben, Elmar Weinhold, and
`Norbert 0. Reich*
`
`476 Gary Braun, Michael Diechtierow,
`(cid:127)
`Stephanie Wilkinson, Falk Schmidt,
`Michael HOben, Elmar Weinhold, and
`Norbert 0 . Reich•
`
`480 Hubert Gaertner, Robin Offord, Paolo Botti,
`(cid:127)
`Gabriel Kuenzi, and Oliver Hartley•
`
`490 Xin Sheng, Xun Guo, Xiao-Min Lu,
`(cid:127)
`Guo-Yuan Lu,* Ying Shao, Fang Liu,* and
`Qiang Xu
`
`Evergeen Ex. 1014
`6 of 20
`
`

`

`Bioconjugate Chem., Vol. 19, No. 2, 2008 9A
`
`Disulfide Cross-Linked Polyethylenimines (PEI) Prepared via Thiolation of
`Low Molecular Weight PEI as Highly Efficient Gene Vectors
`
`499 Qi Peng, Zhenlin Zhong; and Renxi Zhuo
`
`Enhanced Cellular Uptake of Virus-Like Particles through Immobilization
`on a Sialic Acid-Displaying Solid Surface
`
`507 Noriko Ohtake, Kenichi Niikura, •
`(cid:127)
`Tadaki Suzuki, Kena Nagakawa,
`Hirofumi Sawa, and Kuniharu ljiro
`
`Multifunctional Chimeric Proteins for the Sequential Regulation of Neural
`Stem Cell Differentiation
`
`516 Tadashi Nakaji-Hirabayashi, Koichi Kato,
`(cid:127)
`Yusuke Arima, and Hiroo Iwata•
`
`Synthesis, Characterization, Antitumor Activity of Pluronic Mimicking
`Copolymer Micelles Conjugated with Doxorubicin via Acid-Cleavable
`Linkage
`
`525 Yuhan Lee, Sung Young Park,
`Hyejung Mok, and Tae Gwan Park•
`
`Retrograde Delivery of Photosensitizer (TPPp-O-P-GluOH)3 Selectively
`Potentiates Its Photodynamic Activity
`
`532 Mohamed Amessou, Daniele Carrez,
`Delphine Patin, Marianne Sarr,
`David S. Grierson, Alain Croisy,
`Antonio C. Tedesco, Philippe Maillard,* and
`LudgerJohannes
`
`1111n-Labeled Lactam Bridge-Cyclized a.-Melanocyte Stimulating Hormone
`Peptide Analogues for Melanoma Imaging
`
`539 Yubin Miao: Fabio Gallazzi, Haixun Guo,
`and Thomas P. Quinn
`
`Characterization of PLL-g-PEG-DNA Nanoparticles for the Delivery of
`Therapeutic DNA
`
`548 Markus Rimann, Tessa Luhmann,
`Marcus Textor, Barbara Guerino,
`Joelle Ogier, and Heike Hall"
`
`Enhancement of Star Vector-Based Gene Delivery to Endothelial Cells by
`Addition of RGD-Peptide
`
`558 Ayaka Ishikawa, Yue-Min Zhou,
`Nobuaki Kambe, and Yasuhide Nakayama·
`
`TECHNICAL NOTES
`
`Effect of Ligand Density on the Spectral, Physical, and Biological
`Characteristics of CdSe/ZnS Quantum Dots
`
`562 Samuel J. Clarke, C. A. Hollmann,
`(cid:127)
`Faisal A. Aldaye, and Jay L. Nadeau·
`
`Convenient Preparation of 68Ga-Based PET-Radiopharmaceutlcals at
`Room Temperature
`
`569
`
`I. Velikyan, • H. Maecke, and B. Langstrom
`
`Two-Photon Excitation Fluorescence Resonance Energy Transfer with
`Small Organic Molecule as Energy Donor for Bioassay
`
`574
`
`Lingzhi Liu, Genghui Wei, Zhihong Liu,'
`Zhike He, Si Xiao, and Ququan Wang•
`
`(cid:127) Supporting Information is available free of charge via the Internet at http://pubs.acs.org.
`
`• In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the
`paper should be addressed.
`
`Evergeen Ex. 1014
`7 of 20
`
`

`

`10A Bioconjugate Chem., Vol. 19, No. 2, 2007
`
`Aldaye, F. A., 562
`Amessou, M., 532
`Arap, W., 403
`Arima, Y., 516
`Azarin, S. M., 403
`
`Barthelemy, P., 418
`Benini, E., 450
`Bolzati, C., 450
`Bormans, G., 441
`Bosch!, A., 450
`Botti, P., 480
`Braun, G., 476
`
`Carrez, 0., 532
`Gazzola, E., 450
`Gelen, S., 441
`Chitneni, S. K., 441
`Clarke, S. J., 562
`Croisy, A., 532
`Debyser, Z., 441
`de Groot, T., 441
`De Le6n-Rodriguez, L.,
`391
`Oeroose, C., 441
`Diechtierow, M., 470, 476
`Duatti, A., 450
`Dy, E. E., 461
`Estabrook, R. A., 470
`
`Feng, M., 421
`Fox, M. E., 461
`Frechet, J. M. J., 461
`
`Gachet, C., 406
`Gaertner, H., 480
`
`Gallazzi, F., 539
`Gijsbers, R., 441
`Grierson, D. S., 532
`Grinstaff, M. W., 418
`Guerino, B., 548
`Guillaudeu, S. J., 461
`Guo, H., 539
`Guo, X., 490
`
`Haidar, Y. M., 461
`Hall, H., 548
`Hammond, P. T., 403
`Hartley, 0 ., 480
`He, Z., 574
`Hechler, B., 406
`Hollmann, C. A., 562
`Huben, M., 470, 476
`
`ljiro, K., 507
`Ishikawa, A., 558
`Iwata, H., 516
`
`Jacobson, K. A., 406
`Johannes, L., 532
`Kambe, N., 558
`Kato, K., 516
`Kim, Y., 406
`Klutz, A. M., 406
`Kovacs, Z., 391
`Kuenzi, G., 480
`Langer, R., 403
`Langstrom, B., 569
`Lee, C.-C., 428
`Lee, Y., 525
`Lei, Y., 421
`Liu, F., 490
`
`AUTHOR INDEX
`
`Liu, L., 574
`Liu, Y., 428
`Liu, Z., 574
`Lu, G.-Y., 490
`Lu, X.-M., 490
`Lu, Y., 412
`Luhmann, T., 548
`Luo, D., 418
`Maecke, H., 569
`Maillard, P., 532
`Mazumdar, 0., 412
`McIntosh, T. J., 418
`Miao, Y., 539
`Mirtschink, P., 450
`Mok, H., 525
`Mortelmans, L., 441
`Nadeau, J. L., 562
`Nagakawa, K., 507
`Nakaji-Hirabayashi, T., 516
`Nakayama, Y., 558
`Niikura, K., 507
`Offord, R., 480
`Ogier, J., 548
`Ohtake, N., 507
`Park, S. Y., 525
`Park, T. G., 525
`Pasquali, M., 450
`Pasqualini, R., 403
`Palin, 0., 532
`Peng, a., 499
`Pietzsch, H.-J., 450
`Prata, C. A.H., 418
`
`Quinn, T. P., 539
`
`Reich, N. 0 ., 470, 476
`Reineke, T. M., 428
`Rimann, M., 548
`Sarr, M., 532
`Sawa, H., 507
`Schmidt, F., 470, 476
`Shao, Y., 490
`Sheng, X., 490
`Suzuki, T., 507
`Szoka, F. C., 461
`Tang, H., 421
`Tedesco, A. C., 532
`Textor, M., 548
`Uccelli, L., 450
`Vellkyan, l., 569
`Verbruggen, A., 441
`Walther, M., 450
`Wang, a., 574
`Wei, G., 574
`Weinhold, E., 470, 476
`Wilkinson, S., 470, 476
`Wood, K. C., 403
`Xiao, S., 574
`Xu, a., 490
`Yao, L., 421
`Yigit, M. V., 412
`Yu, R., 421
`Zhang, X.-X., 418
`Zhong, Z., 499
`Zhou, Y.•M., 558
`Zhuo, R., 499
`Zou, 8., 421
`
`On the cover. DOTA- peptide conjugates labeled with various metal ions have found applications as targeted MRI contrast
`agents (in vitro MRI detection of a protein- peptide binding event; central theme and top image), in vivo cancer imaging
`agents in animal models (6'Cu-microPET image of a mouse with xenograft; bottom left), and diagnostic/therapeutic agents
`In clinical trials (67Ga scintiscan of a patient showing metastases; bottom right). (See De Le6n-Rodrfguez, L. M., and
`Kovacs, Z., on page 391 .) Images are reproduced with permission.
`
`Digital Object Identifier (DOI): The DOI appears at the bottom of the first page of each article. We suggest that you
`include the DOI in all CCC reporting and document delivery requests. See the masthead of this journal or our Web site
`at http://pubs.acs.org for more details.
`
`1,
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`FEBR ARY 2008
`Volume 19, umber 2
`
`© Copyright 2008 by rhe American Chemical Sociery
`
`Bioconjugate
`Chemistry
`
`REVIEWS
`
`The Synthesis and Chelation Chemistry of DOT A-Peptide Conjugates
`
`Lui M . De Le6n-Rodrfguez and Zoltan Kovac *
`University of Texa Southwe tern Medical Center, Advanced Imaging Re earch Center, 2201 Inwood Road E 4.2,
`Dalla Texa 75390-8568. Received August 28. 2007; Revised Manuscript Received October 11, 2007
`
`Metal complexe of DOT A (1,4,7, I O-tetraazacyclododecane-1,4,7, I 0-tetraacetic acid)-peptide conjugate are
`increasingly used as targeted imaging and therapeutic radiopharrnaceuticals and MRI contrast agents. This review
`cover the bi functional derivative of DOTA, the olution and olid-pha e ynthe i of DOT A- peptide conjugate ,
`their coordination and chelation chemi try. and the biomedical application of variou DOT A- peptide conjugate
`metal complexe .
`
`I TRODUCTION
`
`Peptide-ba ed targeting vectors are becoming increa ingly
`popular for diagnostic and therapeutic applications. They offer
`everal advantages over antibodies including more favorable
`pharmacokinetic and better tumor penetration. Peptide e(cid:173)
`quence that are capable of binding to their target with affinitie
`comparable to tho e of monoclonal antibodie can be generated
`by chemical or phage di play librarie . Affinity election of
`random phage display peptide libraries is a very powerful way
`of identifying novel targeting equence for a receptor from
`billion of peptide (/).
`While diagnostic nuclear imaging i currently dominated by
`99"'Tc, there is no
`ingle i otope that i · perfectly ·uitable for
`the treatment of all cancers. The half-live and energie of the
`available radioi otopes cover a wide range, and the best-suited
`therapeutic isotope ha to be elected according to the ize, type,
`tage of the tumor (2). Due to their favorable nuclear
`and
`propertie . 90Y and radiolanthanide i otope uch a 177Lu and
`
`• Corresponding author. UT Southwe tern Medical Center, Advanced
`Imaging Research Center. 2201 Inwood Road E 4.2. Dallas, TX
`75390-8568 USA. Tel. + I 2146452755. e-mail: zohan.kovac @
`ut outhwc. tcm.edu.
`
`153Sm are frequently cho en for therapeutic applications. Unlike
`their nuclear properties, the chemical behavior of all lanthanide
`and yttrium isotopes is quite similar. The most stable oxidation
`tale of the e element i 3+, and the olution chemistry of the
`i characterized by their trong
`hard, trivalent Ln(IU) cation
`tendency to form complexe with hard donor atom (0, F- ,
`). Their coordination number i u ually 8 or 9, and they
`and
`form thermodynamically very table complexe with polyamino
`polycarboxylate-type ligand
`that have 8 or 9 donor atom
`(Figure I). Among these ligands, the macrocyclk 1,4,7,10-
`tetraazacyclododecane-l,4, 7 I 0-tetraacetic acid (DOT A) (1)
`rand out, a
`it form complexe with Ln(ill) ion with
`. It
`extraordinary thermodynamic tability and kinetic inertne
`should be emphasized that DOTA complexes are significantly
`more inert kinetically than chelate of DTPA (3-6), and although
`both DOTA and DTPA complexes have been u ed a
`imaging
`agent and radiopharrnaceutical , DTPA complexe have been
`shown to di sociate and release metal under physiological
`condition . This may not be of concern in the ca e of an MRI
`exam with Gd-DTPA when clearance of the contrast agent i
`rapid (7. 8), but for targeted radiopharmaceutical application ,
`when the clearance of the complex is slow, DOTA (1) and not
`DTPA (3) should be the ligand of choice.
`
`10.1021/bc700328s CCC: $40.75 © 2008 American Chemical Society
`Published on Web 12/12/2007
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`Leon-Rodriguez and Kovacs
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`....
`
`Figure l. Polyamino polycarboxylate ligand .
`
`2
`
`A large number of DOTA - peptide conjugates have been
`synthesized so far, and ome of them are already in clinical
`applications (9). Some reviews (see references cited in the
`Applications section) have covered the nuclear medicine ap(cid:173)
`plications of DOTA-peptide conjugate based radiopharrnaceu(cid:173)
`tical , while their synthetic and coordination chemi try ha not
`been reviewed in detail. Therefore, thi review i focu ed on
`the ynthe i and chelation chemi try of DOT A -peptide
`conjugate .
`
`BlFUNCTIONAL LIGANDS AND LINKERS
`Construction of a targeted radiopharmaceutical involve the
`covalent attachment (conjugation) of the chelate of the radioac(cid:173)
`tive i otope to the targeting vector using a bifuctional chelator.
`Bifunctional Ligand
`therefore have a reactive functionality,
`usually an aromatic i othiocyanate group or an activated e ter,
`which will react with nucleophilic site (-NH2, -SH, or -OH)
`of the targeting vector (10). A linker may be incorporated
`between the chelator and the targeting vector to influence the
`pharmacokinetic properties of the conjugate (I 0). Hydrocarbon,
`PEG, or polypeptide Linkers can alter the pharmacokinetics and
`biodistribution by changing the overall charge and hydrophilicity
`of the drug (// - 13). Peptide-ha ed linker can al o incorporate
`specific metabolizable equence . An elegant method based on
`an on-demand cleavable" peptide sequence has been developed
`recently to minimize the radiation delivered to normal tis ues.
`A short peptide linker (rqYKYkf) identified from combinatorial
`peptide libraries was incorporated between the DOT A moiety
`and the targeting vector (ChL6 monoclonal antibody) of a
`targeted radiophannaceutical. This equence can pecifically be
`cleaved by the thrombolytic enzyme TNKa e that i re tricted
`to the blood pool. After adequate tumor localization of the drug
`the enzyme was administered intravenously. Since the enzyme
`tayed in the blood pool , it released the radioactive DOT A
`chelate only from the circulating radiophannaceutical. The rapid
`renal clearance of the small-molecule DOTA complex re ulted
`ue (14).
`in dimini bed radiation do e to normal ti
`
`SYNTHESIS OF DOTA PEPTIDES
`Conjugating DOTA to Antibodies. Initial trategie to attach
`DOT A to peptide emerged from earlier methodologie devel(cid:173)
`oped for antibody labeling. Protein are most commonly
`modified on the primary amino groups of lysine sidechains. Due
`to their polar and hydrophilic nature, these are usually located
`on the surface of proteins and thus are easily functionalized.
`p-1 othiocyanatobenzyl DOTA (4) was the first DOTA bifunc(cid:173)
`tional chelator developed for protein labeling and continue to
`be the most widely used DOTA bifunctional for antibody
`labeling (15- 17). It i water- oluble and reacts easily with ly ine
`side chains in lightly ba ic olution (pH 8-9) (18). Encouraged
`by the uccess of this bifunctional, other DOTA derivative with
`an aromatic SCN group have been developed (S-9) (Figure
`2), but the e do not show significant improvements in label.ing
`yield or in vivo tability (19-21).
`The synthesis of these SCN derivative , especially the
`backbone ub tituted compounds, is fairly lengthy and tediou ,
`and o other approache based on the in
`itu activation of the
`much more readily available DOTA have been developed. A
`imple procedure is based on the activation of one of the acetic
`
`acid sidearm of DOT A with isobucyl chloroformate in the
`presence of tetramethyl guanidine to form a mixed anhydride
`derivative (22). lo situ preparation of HS and ulfo-NHS e ters
`of DOTA were also reported (23, 24). Although protein were
`ucce fully labeled with the e mixture under very mild
`condition , thi approach has two disadvantages. Fir t, in order
`to uppress the cro -linking due to the activation of more than
`one acetate sidearm, a large excess of DOT A has to be used.
`Second, these activated esters are quite sensitive to hydrolysis,
`and thus the labeling result are unpredictable.
`Both problem can be overcome by u ing preformed
`DOTA - NHS ester (10) (Figure 3) that can be
`tandardized
`before conjugation (25). Since the preformed DOTA-NHS is
`ynthesized using a selective protection-functionalization(cid:173)
`deprotection approach to ensure that only one of the acetate
`ideanns be activated, the formation of cro s-linked
`ide
`products during bioconjugation is eliminated. While DOT A-NHS
`can label proteins at neutral pH under milder condition
`than
`p-i othiocyanatobenzyl DOTA (26), it hydrolyze
`rapidly in
`water (27). To minimize the hydrolysis problems mentioned
`above, everal DOTA phenolic-activated e ter derivatives were
`prepared (11 -18) from the corresponding phenol, DOT A, and
`EDC or DCC, followed by HPLC purification (28) (Figure 3).
`Some of these derivative showed higher hydrolitic tabilitie
`(11 14, and 17) compared to DOTA- HS (10) (28), a well
`as good labeling efficiencies with albumin (29).
`Although in proteins the lysine side chain amino groups are
`the most cornrnonly labeled conjugation of DOT A through the
`thlol group of cy tein residue with DOT A maleimide deriva(cid:173)
`tives under relatively mild condition (pH ~ 7) ha al o been
`reported (30, 31).
`Solution-Phase Conjugation. In olution, peptide and
`peptide derivatives can be conjugated to DOT A in the ame
`manner as antibodies using water-soluble bifunctional ligand .
`For example, an E. coli heat- table peptide analogue was
`conjugated in aqueou olution at pH 8.5 with the i othlocyanate
`derivative 6 through an aminocaproic acid linker (32). In
`itu
`prepared DOTA-NHS ester was also used to conjugate DOTA
`to the N-terminus of protected octreotide and octreotate deriva(cid:173)
`tive
`in a water-DMF mixture in which both DOTA and the
`protected peptide were soluble (33, 34). Thi method can be
`e pecially advantageou when the peptide i not available in
`the protected form a was the ca e with certain cyclic ROG
`peptide derivatives. Good coupling yields (up to 79%) of DOTA
`to RGD peptide monomers and multimers were obtained
`by using an excess of a mixture of DOT A/EDC/sulfo-NHS
`(10:5:4). Activation of DOTA in the form of the sulfo-NHS
`ester was allowed to run for 30 min at 4