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Synthesis 2003(17): 2647-2654  
DOI: 10.1055/s-2003-42475
PAPER
© Georg Thieme Verlag Stuttgart · New York

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

Carlos A. M. Afonsoa, V. Santhakumarb, Alan Loughb, 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;
Further Information

Publication History

Received 8 November 2002
Publication Date:
21 November 2003 (online)

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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

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15

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).

16

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, Dc = 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].

17

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