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A Mild Method for Regioselective Labeling of Aromatics with Radioactive Iodine
Mads H. Rønnest,[a,b] Felix Nissen,[c] Palle J. Pedersen,[a] Thomas O. Larsen,[b] Walter Mier,[c] and Mads H. Clausen*[a]
Keywords: Isotopic labeling / Radiochemistry / Medicinal chemistry / Iodine / Silanes / Thallium
A novel technique to label ortho-, meta-, and para-trimethylsilyl-substituted aryl substituents with radioactive iodide is described. The method takes advantage of the ipso-directing and activating properties of trimethylsilyl substituents on the arenes. The method was demonstrated on a griseofulvin analogue with promising anticancer properties and on lidocaine, a widely used local anesthetic drug. Treatment of a trimethylsilyl precursor with Tl(OCOCF3)3 followed by Na125I consistently afforded radioactive purities over 95 % in all cases.
Methods for labeling of organic molecules with radioactive isotopes of iodine, such as 123I, 125I, and 131I, are important for medical applications. Iodinated drug candidates are used for screening antibodies in radioimmunoassays (RIAs) and in preclinical ADME (absorption, distribution, metabolism, and excretion) studies. Owing to emission of gamma radiation, iodinated radiopharmaceuticals are also used in scintigraphic imaging for disease diagnostics, for example, in the localization of tumors by use of meta-iodobenzylguanidine (MIBG) labeled with 123I. The iodination of aromatics can be achieved by different methods, and the choice of labeling method depends very much on the substrate. Many of the most widely used labeling techniques involve the use of oxidizing agents such as chloramine T, iodogen, and lactoperoxidase; however, these methods suffer from low selectivity and are restricted to activated aromatics. The regioselectivity (labeling position) can be hard to control and to some extent so can the degree of labeling (the number of iodine atoms introduced). Depending on the substrate, this can result in the formation of numerous side products,[5b] which then have to be purified, often by tedious chromatographic methods. Further[a] Department of Chemistry, Center for Nanomedicine and Theranostics, Kemitorvet 207, 2800 Kgs. Lyngby, Denmark Fax: +45-4593-3968 E-mail: firstname.lastname@example.org Homepage: http://www.organic.kemi.dtu.dk/Research/mhcl [b] Department of Systems Biology, Center for Microbial Biotechnology, Søltofts Plads, Building 221, 2800 Kgs. Lyngby, Denmark [c] Department of Nuclear Medicine, University Hospital Heidelberg, 69120 Heidelberg, Germany Supporting information for this article is available on the WWW under http://dx.doi.org/10.1002/ejoc.201300419. 3970
more, many iodination methods are only applicable to activated aromatics, such as tyrosine in labeling of peptides and proteins. Therefore, there is a need for a broader collection of selective iodination techniques that can be performed under mild conditions. Our interest in iodination techniques arose during our search for potent analogues of the natural product griseofulvin (1) that can inhibit centrosomal clustering in cancer cells. GF-15 (compound 2, Figure 1) was identified as one of the most active analogues, and for in vivo biodistribution and half-life studies we became interested in 125I labeling of 2. As the activities of the three iodinated isomers of 2 (i.e., 8–10, see Scheme 1) in a phenotypic whole-cell assay for multipolarity are markedly different (data not shown), we found it crucial to apply a method that would afford a single regioisomer and thus enable us to access the ortho- (i.e., 8), meta- (i.e., 9), and paraiodo (i.e., 10) isomers.
Figure 1. Structures of griseofulvin (1), its analogue GF-15 (2), griseofulvic acid (3), and lidocaine (4).
McKillop et al. have introduced a method for the iodination of small aryl-containing organic molecules by the use of Tl(OCOCF3)3 and iodide. Mechanistically, the thallated aromatics are formed by direct electrophilic metalation of the aryl ring with Tl(OCOCF3)3, after which ligand exchange with iodide followed by ipso substitution creates the aryl–iodine bond. Whereas the reaction of the
Eur. J. Org. Chem. 2013, 3970–3973
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and 7 into the iodo products was 1. The formation of only the ipso-substituted product was confirmed by comparison of the NMR spectra and the LC–MS data for iodides 8–10. the conversion to the iodo product was Ͻ20 %. The labeling is typically performed in pure trifluoroacetic acid (TFA). Therefore. 5. which presumably is a consequence of the meta-substitution pattern in 6. no conversion of 2 was observed. NaI. incorporation of radioactive iodide into griseofulvin analogue 2 would be possible via the corresponding thallated intermediates.) was added. MeCN/TFA (7:3). but when milder conditions [Tl(OCOCF3)3 in MeCN/TFA. To further elucidate the scope of the technique.6-dimethylaniline. as radioactive iodide is used in substoichiometric amounts. Lidocaine (4) is a widely used local anesthetic drug. Figure 1) was also investigated. which means that deactivated aromatics can be challenging to label. 9.. the iodination of 7 (para-TMS) was tested with a lower amount of TFA (10 and 20 % TFA in MeCN) but still with the use of 2 equiv. Eur.17] In the case of substrates that cannot tolerate these conditions. As evident from the chromatograms in Figure 2. and this underlines that protodesilylation does not occur under the conditions. respectively. It was also investigated how the reaction was affected by using lower amounts of Tl(OCOCF3)3 (0. depending on the substrate and conditions. such as MeCN.9 equiv. 5–7) were dissolved in MeCN/TFA (7:3) and then subjected to Tl(OCOCF3)3 (2 equiv. iodination of lidocaine (4. whereas at 20 % TFA. and under these conditions. and iodination of this drug has previously been explored by El-Moselhy et al.[16. and 10 were isolated by column chromatography in 44. the iodo product was only observed in trace amounts (LC–MS analysis).) for 30 min after which NaI (4 equiv.eurjoc. in which the radioactive iodine was the limiting reagent. Chem... TMS analogue 11 (see Scheme 2) of lidocaine (4) was synthesized in three steps from commercially available 4bromo-2. such as 3-trimethylsilyltoluene. The three mono-TMS-analogues of griseofulvin (i. we went on with 125I labeling. At 10 % TFA. a regioselective variant of the iodination method was published by Bell et al. the radiochemical purity (RCP) by HPLC for all three radioiodinations was Ͼ95 %.). 7:3] were used. MeCN/TFA (7:3). 5–7. for use in labeling with radioactive iodide the low conversion in the electrophilic aromatic thallation is not critical. hydrolysis to form griseofulvic acid (3. and the reaction mixtures were stirred for 2 min and then quenched by the addition of a saturated aqueous solution of Na2CO3. J. and 80 % yield. addition of a cosolvent. The identity of the labeled compounds was confirmed by coelution with nonradioactive reference compounds in the HPLC analyses (detected in the UV channel).) in MeCN/TFA (7:3). RCP = radiochemical purity. the introduction of the TMS group increases the reactivity of the aromatic system. The low isolated yield of 9 is due to low conversion of starting material 6 into the thallated intermediate. Results and Discussion In continuation of the work by McKillop et al. Weinheim 3971 . addition of a cosolvent also lowers the reactivity. Na125I.Regioselective Labeling of Aromatics with Radioactive Iodine Scheme 1. the initial thallation can yield mixtures of the three possible regioisomers. whereby less activated aryls can be thallated and iodinated. For this substrate. Gratifyingly. and HPLC analysis was performed after 30 s. Tl(OCOCF3)3 and an excess amount of NaI. Figure 1) took place within minutes. whereas some unreacted starting material was also detected. respectively. 3970–3973 Initially. undergo ipso substitution when treated with Tl(OCOCF3)3. However. Iodinated products 8. aryl thallium(III) bis(trifluoroacetate) intermediate with iodide affords solely the ipso-substituted product. can usually prevent substrate degradation. The TMS-benzyl analogues of griseofulvin (i. which were made through a conventional route from the corresponding iodobenzyl alcohols (see the Supporting Information). (b) Tl(OCOCF3)3. In complete absence of Tl(OCOCF3)3. and 7 in MeCN/ TFA (7:3) after which the thallation was initiated by addition of Tl(OCOCF3)3 (0.7–3. Scheme 1) were synthesized from the corresponding TMSbenzyl alcohols by using known procedures (see the Supporting Information). no conversion of the starting material took place. 6. whereby it becomes the limiting reagent anyway (vide infra). to the best of our knowledge. motivated by the work of Bell et al. we took adwww. and 7 (see Scheme 1). 6. Org. whereas none of the other regioisomers are formed above trace levels.org © 2013 Wiley-VCH Verlag GmbH & Co. Labeling of griseofulvin analogues with 127I and 125I. the labeling procedure was tested with nonradioactive iodide. and this demonstrates the necessity of TFA for the reaction to occur. Furthermore. With the promising results from the iodination with 127I in hand. KGaA.e. Na125I (1. we hypothesized that by use of TMS-aryls 5. When we exposed griseofulvin analogue 2 to the reaction conditions [Tl(OCOCF3)3 in neat TFA].4 ϫ 10–4 equiv. Furthermore. The labeling was performed by dissolving TMS precursors 5. Additionally. only been used scarcely and only with nonradioactive iodide. However. LCMS analysis of the reaction mixtures showed that the corresponding iodides were formed as the only product. 8. Reagents: (a) Tl(OCOCF3)3. The reaction mixture was stirred for 2 min at 20 °C. the identity and uniformity of the products were verified by their retention times in HPLC analyses. 6.e. They discovered that trimethylsilyl (TMS)-substituted aryls. and 20 %. 2013. The application of this iodination technique has.) was added. the conversion of 5.5 equiv.
23] which are typically converted into the corresponding aryl iodide by treatment with NaI and chloramine T (or another oxidant such as peracetic acid) in an acidic aqueous environment. Clausen et al. this regioselective iodination method was found to be very convenient. mation. Supporting Information (see footnote on the first page of this article): Experimental procedures..5 equiv. After 2 min of stirring at 20 °C. and Tl(OCOCF3)3 should be handled with care in a well-ventilated fume hood. and 10. 1–2 MBq. Radiochemical purities Ͼ95 % were achieved in all cases. and this verifies that under mild reaction conditions the TMS group is crucial for the reactivity of the aromatic ring. for details). MeCN/TFA (7:3). respectively. and ingestion. KGaA. The test iodination with Na127I (0. 0. (b) Tl(OCOCF3)3. 125 Figure 3. Weinheim . and NMR spectra. Overall. Subjection of lidocaine (4) with no TMS group to the iodination conditions gave no iodide incorporation. LC–MS data. skin contact. J.SHORT COMMUNICATION M.[5a. 3970–3973 The use of TMS-aryls as precursors for radioiodination has been reported previously. Experimental Section Typical Procedure for 125I Labeling: The TMS-analogue (70 nmol) was dissolved in MeCN/TFA (7:3. HPLC analysis was performed. NaI.g. NaI and tert-butyl hypochlorite or H2O2 in acetic acid at 60 °C) and long reaction times (Ͼ30 min) are needed to obtain reasonable conversion.org © 2013 Wiley-VCH Verlag GmbH & Co. and 7 into corresponding iodides 8. H. Scheme S1. Hence. The conditions used are superior to those previously reported. 35 µL).33 m. Eur. see Supporting Information). Safety Note: Thallium compounds are extremely toxic to inhalation. vantage of one of the mild methods available for the introduction of a TMS group: the palladium-catalyzed crosscoupling of an aryl halide with hexamethyldisilane (see the Supporting Information).5 equiv. Another popular method for iodination of aromatics through ipso substitution of an activator group is by use of arylstannanes. 2013. Conclusions We were successful in applying the electrophilic aromatic thallation for activation of TMS-aryls in the radioiodination of a potent griseofulvin analogue and lidocaine.[5. which minimizes the time in which the molecules are subjected to the acidic reaction conditions. which demonstrates that H2O is tolerated in the reaction (see the Supporting Information for LC–MS analysis). 35 nmol) was added. MeCN/TFA (7:3). Toxicity is cumulative. The iodination also occurred if MeCN/TFA/H2O (6:3:1) was used as the solvent mixture. Tl(OCOCF3)3 in MeCN/TFA (7:3) gave desired iodinated lidocaine 12 (full conversion by LC–MS.) with the use of 0. especially for molecules with low water solubility and for substrates that are sensitive to highly acidic environments (such as neat TFA) or oxidizing reagents. 6. Na125I.eurjoc. 15 µL.21] However the reported reaction conditions are rather harsh (e. Org. Figure 2. Reagents: (a) Tl(OCOCF3)3. 12– 24 pmol) was added. 2. The need for new procedures that can transform TMS-aryls into their corresponding iodine analogue was recently demonstrated in the attempted iodination of a tomography tracer for the histamine H3 receptor. whereas when neat TFA was used as the solvent. and we envision that this procedure will be a strong alternative to the use of other labeling strategies. iodination was observed. Chem. the use of TMS-aryls and Tl(OCOCF3)3 is an attractive alternative to arylstannanes. as the iodination proceeds at ambient temperature and in a less acidic environment. Iodination of TMS-lidocaine analogue 11. after which a solution of Tl(OCOCF3)3 in MeCN/TFA (7:3. Performing the reaction with 11 and Na125I gave the desired incorporation of 125 I with high radiochemical purity as shown for two independent experiments in Figure 3. 9.1 µL. for which the use of standard reagents such as chloramine T and peracid resulted in less than 10 % labeling (see the Supporting Infor3972 www. Scheme 2. HPLC analysis (radio channel) of the 125I labeling of lidocaine analogue 11. It is also noteworthy that the radioiodination with thallium trifluoroacetate is performed for only 30 s. The reaction mixture was stirred for 2 min at 20 °C and then an aqueous solution of Na125I (ca. HPLC analysis (radio channel) of the I labeling of griseofulvin analogues 5.
R. Fowler. Org. K. 1969. K. 170–190. Wagner. J. S. Clin. Lett. 2000. R. E. 685–689. Toxicol. Hanson. J. 2013. U. L. K. H. Shirakawa. Res. Rev. b) V. Commun. 575–590. P. C. 2000. A. G. J. Chichester. 1980. B. h) M. E. Kurahashi. M. Yevich. S. A. b) S. J. Ho.  J. M. Ogan. M. 9.  M. J. 3483–3486. B. UK. Bubeck. D.  a) P. Fowler. H.  E. Probes and Contrast Agents (Ed. 1980. De Spiegeleer. Mier. b) A. Int. Summerfield. T. Lehrmann. D. Labelled Compd. Dischino. J.  A. Clausen. Chem. J. 129–137. 850. 1895–1896. Jeffrey. S. Peremans. Markworth. Baille. Eldawy. 2003. J. J. Am. Hellman. 97. Organomet. E. Soc. S.  a) D. Rønnest. e) D. P. Rebacz. C. W. J. Larsen. J. KGaA. Yalow. f) L. D. M. 987–994. F. 1924–1932. Hunt. Rønnest. J. Biochem. Anderson. J. M. B. Burvenich. The Netherlands. K. Med. McKillop. 43. Krämer. 72. D. Hunter. 35. G. 4845–4850. C. L. Res. J. c) R. b) M.  S. R. Springer. J. R. A. Larsen. Valliant. O. Rønnest. Clin. A. F. 1974. J. 2686–2718.  a) R. 1963. O. Hunt. 1969. Chem. Mier. b) W. Hubers. Biochem. H. N. d) H. A. Krämer. T. 2423–2426. Nissen. Adelstein. Buchwald. M. Chem. 1982. Van der Heide. Haberkorn. Endocrinol. 2007. J. 5. M. 1998. Eersels. Wilbur. J. 2013 Eur. G. Hunt. Goldsmith. C. J. S. S. J. M. Konotop. Seitz.  a) D.. Cancer 1996. T. Dordrecht. Mier. Haberkorn. 82. 2011. 2009. Lay. R. R. D. J. H. Mazieere.)  a) S. Robey. 1978. J. El-Kersh. Biophys. Boot. J. W. H. 65–71. Cancer Res. J. S. Berson. Commun. Chem. A. Cambridge. 2012. Springer. A. D. 45. S. 49–55. Valdes Olmos. L. 80. Mukherjee. Speck Jr. R. S. Res. Anderhub. E. J. H. Koehler. A. S. Kaltsas. D. O’Brien Jr. Mattson. T. Coenen. Breitkreutz. in: Handbook of Nuclear Chemistry. 3785–3788. McGillivray. E. Cancer Res. F. J. the Danish Cancer Society and Karen Krieger Fonden (support to P. J. Parker. Eisenhut. Biochem. McGillivray.Regioselective Labeling of Aromatics with Radioactive Iodine Acknowledgments This research was supported by the Danish Research Council (ref. J. 1 1985. 2006. W. H. S. C. J. Soc. M. Barder. D. H. 2007. Labelled Compd. C. O. Nieuwenhuizen. Baert. 274-07-0561. J. J. 54. Radiopharm. 21. S. H. Am. El-Moselhy. M. A. 15. Hiyama. J. Hunt. F. Zelesko. L. Fowler. Slegers. I. L. Kienzle. H. Oxford. 114–123. Med. 1970. M. Radioiodination Reactions for Radiopharmaceuticals. M. Champion. 2008. C. A. T. 93. M. 849–857. Chem. F. 93. Eisenhut. 2003. H. K. c) A. 3391–3394. Anderson. McKillop. P.  J.  a) G. Greenwood. Emeis. 1998. J. Res. Anderhub. 299–305. MoBiTech.. H. Putignano. Moerlein.  H. O. 4841–4844. Rønnest. Pinhey. Boesen. W. Gorgas. H.  A. Hull. Redvanly). 1969. Molecules 2010. Perkin Trans. C33–C36. Rothschild. M. L. G. Chem. b) S. C. Bauman. H. J. B.. J. J. Löffler. Scott. N. Expert Opin. Radiopharm. Larsen. 1997. Eur. Labelled Compd. A. 2012. Wiley. Chem. S. 2013 Published Online: May 14. b) B. H. J. 4. Tetrahedron Lett. g) M. 9. 1971. 3342–3347. E. H. Clausen. McKillop. Taylor. Hunt. Satta. McKillop. J. H. A. Vanbree. 13. F. Besser. G. C. Coenen. Instrum. G. C. P. Podar. M. 127–133. McNeill. Invest. F. Radiopharm. 3913–3922. Finn. b) F. Simonart. 89. M. Lowe. Robichaud. T. and the German Federal Ministry of Education and Research (BMBF. Hoefnagel.  D. L. J. Labelled Compd. Nicholl. W. 194. Nyberg. T. J. Glover. 1975.). Britton. 2012. H. Commun. Hunter. 495–496.  a) T. Biochem. Vermond. Chromatogr. Krämer. U. Hull.: M. S. J. Tonneson. Taal. Clausen. b) E. Chem. Denmark. 2011.  a) M. A. J. 2012. R. G. Appl. 49. J. Biochem. McQuarrie. K. C. Nucl. 1171– 1188. Kienzle. Greenwood. C. Mohammed. J. A. Soc. E. D. Labelled Compd. 38. Chem. Received: March 20. Kallemeyn. 67. Braddock).. P. M. M. Tetrahedron Lett.  a) M.  B. Fraker. A. 8. Goldschmidt. A. W. 15 Biomedical Imaging: The Chemistry of Labels. D. M. Kurver. Med. Wuest. Yoshida. b) S. M. F. 33. Raab. 6342– 6350. S. J. J. Gagnon. Org. 1939. P. McGillivray. Org. B. 1941–1947. Taylor. and M.). Bell. Radiopharm. P. Taylor. E. Chem. D. A. Radiopharm. Zelesko. E. C.  a) W. J. 32. 15. 1971. Taylor. Nature 1962. in: RSC Drug Discovery Series No. Just. K.  a) A. 3970–3973 © 2013 Wiley-VCH Verlag GmbH & Co. Bodé. Royal Society of Chemistry. Med. H. T. McKillop. H.  a) E. R. J. 5374–5385. Sternhell. M. D. Weinheim www. Seevers. J. S. Terp. Stone. b) A. W. S. J. Newerly. 83–88. Eisenhut. Clausen. Welch. H. Drug Discovery 2009. A. P. W. J.eurjoc. 186. K. J. Pimlott. b) C. A. C. P. J. 2427–2430. Gross.: M. Chem. Ho. Counsell. W. 55. Med. Lett. M. S. 240–248. Semin. A. S. c) S. Travis. Mertens. A. 1982. F. Chem. J. J. T. F. C. H. 674–677. 241–257. Herscheid. in: Handbook of Radiopharmaceuticals: Radiochemistry and Applications (Eds. 10. W.  T. 652–660. 1956. 5. 77– 88. B 2007.org 3973 . Hede. M. J. E. Raab. Marchalonis. support to F. B. Biophys. A. Grossman. 54. 2005. J. Bourdoiseau. G. B 1986. H. Kalman. Fruehaul. Leber. 163–170. O. Larsen. Tetrahedron Lett. 213–220. Zelesko. Radiat. A. E. Vergote. 113. Monson. A. 48. Krämer. Donovan. M. 2002. H. Med. 52. support to M. D. J. 125–152. 10. 24. Raistrick. Chem. Mohammed. J. Berlin. Rebacz. 207–218. Hayden. 19. G. Nucl. J. B. Organomet. D. and M.