An Expedient Synthesis of Cationic Rhodamine Fluorescent Probes Suitable for Conjugation to Amino Acids and Peptides

Carlos A. M. Afonso; V. Santhakumar; Alan Lough; Robert A. Batey

doi:10.1055/s-2003-42475

PAPER

An Expedient Synthesis of Cationic Rhodamine Fluorescent Probes Suitable for Conjugation to Amino Acids and Peptides

Carlos A. M. Afonso^a, V. Santhakumar^b, Alan Lough^b, Robert A. Batey*^b
^a Department of Chemistry, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2825 Monte de Caparica, Portugal
e-Mail: cma@dq.fct.unl.pt;
^b Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario, M5S 3H6, Canada
Fax: +1(416)9785059; e-Mail: rbatey@chem.utoronto.ca;

Abstract

Alle Artikel dieser Rubrik

Abstract

Rhodamine 19 benzyloxycarbonylmethyl ester bromide 8 and rhodamine 19 4-chloromethyl-1-phenylmethyl ester chloride 12 act as precursors to cationic fluorescent probes. Molecules containing free amine or carboxylate functional groups, respectively, can be attached in a one-step procedure, yielding the desired probes without the need for chromatographic purification. As a proof of concept the method was applied to the attachment of amino acid and dipeptide residues through either the N- or C-termini. The precursor molecules 8 and 12 are readily synthesized from the inexpensive, commercially available dye rhodamine 6G.

Key words

fluorescent probes - rhodamines - amino acid derivatives - peptides

Volltext

Referenzen

References

<A NAME="RM04802SS-1">1</A> Valeur B. Molecular Fluorescence: Principles and Applications Wiley-VCH; Weinheim: 2002.

<A NAME="RM04802SS-2A">2a</A> Haugland RP. Handbook of Fluorescent Probes and Research Products Molecular Probes, Inc.; Eugene: 2001. and references cited therein

<A NAME="RM04802SS-2B">2b</A> Probe Design and Chemical Sensing, In Topics in Fluorescence Spectroscopy Vol. 4: Lakowicz JR. Plenum Press; New York: 1994.

<A NAME="RM04802SS-2C">2c</A> Scala-Valéro C. Doizi D. Guillaumet G. Tetrahedron Lett. 1999, 40: 4803

<A NAME="RM04802SS-3A">3a</A> Rahavendran SV. Karnes HT. Anal. Chem. 1996, 68: 3763

<A NAME="RM04802SS-3B">3b</A> Soper SA. McGown LB. Warner IM. Anal. Chem. 1994, 66: 428R

<A NAME="RM04802SS-4A">4a</A> Finney NS. Curr. Opin. Drug Discovery Dev. 1998, 1: 98

<A NAME="RM04802SS-4B">4b</A> Lescrinier T. Hendrix C. Kerremans L. Rozenski J. Link A. Samyn B. Aerschot AV. Lescrinier E. Eritja R. Beeumen JV. Herdewijn P. Chem.-Eur. J. 1998, 4: 425

<A NAME="RM04802SS-5A">5a</A> Mitchison TJ. Sawin KE. Theriot JA. Gee K. Mallavarapu A. Caged Compounds, In Methods in Enzymology Vol. 291: Marriott G. Academic Press; New York: 1998. p.63

<A NAME="RM04802SS-5B">5b</A> Boturyn D. Boudali A. Constant J.-F. Defrancq E. L’homme J. Tetrahedron 1997, 53: 5485

<A NAME="RM04802SS-5C">5c</A> Harapanhalli RS. Roy AM. Adelstein SJ. Kassis A. J. Med. Chem. 1998, 41: 2111

<A NAME="RM04802SS-5D">5d</A> Mayer A. Neuenhofer S. Angew. Chem., Int. Ed. Engl. 1994, 33: 1044

<A NAME="RM04802SS-5E">5e</A> Kojima H. Hirotani M. Urano Y. Kikuchi K. Higuchi T. Nagano T. Tetrahedron Lett. 2000, 41: 69

<A NAME="RM04802SS-6">6</A> Davie E. Morris JH. Smith E. Org. Mass Spectrom. 1974, 763

<A NAME="RM04802SS-7">7</A> Cai SX. Zhang H.-Z. Guastella J. Drewe J. Yang W. Weber E. Bioorg. Med. Chem. Lett. 2001, 11: 39

<A NAME="RM04802SS-8A">8a</A> Bain AJ. Chandna P. Butcher G. Bryant J. J. Chem. Phys. 2000, 112: 10435

<A NAME="RM04802SS-8B">8b</A> Byassee TA. Chan WC. Nie S. Anal. Chem. 2000, 72: 5606

<A NAME="RM04802SS-8C">8c</A> Fang X. Tan W. Anal. Chem. 1999, 71: 3101

<A NAME="RM04802SS-8D">8d</A> Mandala M. Serck-Hanssen G. Martino G. Helle KB. Anal. Biochem. 1999, 274: 1

<A NAME="RM04802SS-8E">8e</A> Nie S. Chiu DT. Zare RN. Science 1994, 266: 1018

<A NAME="RM04802SS-8F">8f</A> Matsumoto Y. Sasaoka N. Tsuchida T. Fujiwara T. Nagao S. Ohmoto T. J. Neurooncol. 1992, 13: 217

<A NAME="RM04802SS-8G">8g</A> Choi KJ. Turkevich LA. Loza R. J. Phys. Chem. 1988, 92: 2248

<A NAME="RM04802SS-8H">8h</A> Kuzela S. Joste V. Nelson BD. Eur. J. Biochem. 1986, 154: 553

<A NAME="RM04802SS-9">9</A> Heilporn S. Broeders F. Daloze D. Braekman JC. Boeynaems JM. Bull. Soc. Chim. Belg. 1994, 103: 309

<A NAME="RM04802SS-10">10</A> Gallo EA. Gellman SH. J. Am. Chem. Soc. 1993, 115: 9774

<A NAME="RM04802SS-11">11</A> For a similar linking strategy for peptides to fluoresceins and their application as fluorescent reporters, see: Chen C.-A. Yeh R.-H. Lawrence DS. J. Am. Chem. Soc. 2002, 124: 3840

<A NAME="RM04802SS-12A">12a</A> Adamczyk M. Grote J. Bioorg. Med. Chem. Lett. 2000, 10: 1539

<A NAME="RM04802SS-12B">12b</A> Cincotta L, and Foley JW. inventors; U.S. Patent 4,290,955. See also:

<A NAME="RM04802SS-13A">13a</A> Mayer U, and Oberlinner A. inventors; U.S. Patent 4,647,675.

<A NAME="RM04802SS-13B">13b</A> Arnost MJ, Meneghini FA, Palumbo PS, and Stroud SG. inventors; U.S. Patent 4,900,686.

<A NAME="RM04802SS-13C">13c</A> Mayer U, and Oberlinner A. inventors; U.S. Patent 4,935,059.

<A NAME="RM04802SS-13D">13d</A> Haugland RP, Singer VL, and Yue ST. inventors; U.S. Patent 6,399,392.

<A NAME="RM04802SS-14">14</A> Wakamiya T. Saruta K. Yasuoka J.-I. Kusumoto S. Bull. Chem. Soc. Jpn. 1995, 68: 2699

The relative quantum yields at 495 nm of derivatives 10, 13 and 15 were 1.26, 1.03 and 0.95 × that of rhodamine 6G respectively (see Ref. [18] for the method used).

The crystals of compound 14 were obtained by crystallization from Et₂O-EtOH: C₄₁H₄₆IN₃O₇, M = 819.71, triclinic, space group P-1, a = 12.6230(2), b = 13.4724(3), c = 13.5600(3) Å, α = 101.865(1), β = 107.003(1), γ = 113.602(1)°, V = 1878.77(7) Å³, Z = 2, D_c = 1.449 gcm^-3, µ = 0.907 mm^-1, F(000) = 844, crystal dimensions 0.35 × 0.32 × 0.28 mm³. The intensities of 28224 reflections were measured on a Nonius Kappa-CCD diffractometer (MoKα radiation, T = 100.0(1) K, 4.19 < θ < 30.48, 10822 unique reflections). The structure was solved and refined using the SHELXTL package (see Ref. [17] ). Non-hydrogen atoms were assigned anisotropic thermal parameters. Hydrogen atoms were included in calculated positions and treated as riding atoms. The refinement which included 469 parameters, converged with R1[I>2σ(I)] = 0.0375 (for 8449 reflections with I>2σ(I)) and wR2 (all unique data) = 0.0970. Atomic coordinates and further crystallographic details have been deposited at the Cambridge Crystallographic Data Centre, deposition number CCDC-191818, and copies of this data can be obtained in application to CCDC, 12, Union Road, Cambridge CB2 1EZ, UK. [Fax: +44(1223)336033; E-mail: deposit@ccdc.cam.ac.uk].

Sheldrick, G. M. SHELXTL/PC Version 5.1, Windows NT Version, Bruker AXS Inc., Madison, USA.

<A NAME="RM04802SS-18">18</A> Parker CA. Photoluminescence of Solutions Elsevier; Amsterdam: 1968.

<A NAME="RM04802SS-19">19</A> Kohno Y. Narasaka K. Bull. Chem. Soc. Jpn. 1995, 68: 322