Top

European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-04-2008 | Original article

Pharmacological evaluation of [¹²³I]-CLINDE: a radioiodinated imidazopyridine-3-acetamide for the study of peripheral benzodiazepine binding sites (PBBS)

Authors: Filomena Mattner, Karine Mardon, Andrew Katsifis

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 4/2008

Abstract

Purpose

The study aims to evaluate the iodinated imidazopyridine, N′,N′-diethyl-6-Chloro-(4′-[¹²³I]iodophenyl)imidazo[1,2-a]pyridine-3-acetamide ([¹²³I]-CLINDE) as a tracer for the study of peripheral benzodiazepine binding sites (PBBS).

Materials and methods

In vitro studies were performed using membrane homogenates and sections from kidney, adrenals, and brain cortex of Sprague-Dawley (SD) rats and incubated with [¹²³I]-CLINDE. For in vivo studies, the rats were injected with [¹²³I]-CLINDE. In competition studies, PBBS-specific drugs PK11195 and Ro 5-4864 and the CBR specific drug Flumazenil were injected before the radiotracer.

Results

In vitro binding studies in adrenal, kidney, and cortex mitochondrial membranes indicated that [¹²³I]-CLINDE binds with high affinity to PBBS, K _d = 12.6, 0.20, and 3.84 nM, respectively. The density of binding sites was 163, 5.3, and 0.34 pmol/mg protein, respectively. In vivo biodistribution indicated high uptake in adrenals (5.4), heart (1.5), lungs (1.5), kidney (1.5) %ID/g at 6 h p.i. In the central nervous system (CNS), the olfactory bulbs displayed the highest uptake; up to six times the activity in blood. Pre-administration of unlabeled CLINDE, PK11195 and Ro 5-4864 (1 mg/kg) reduced the uptake of [¹²³I]-CLINDE by 70-55% in olfactory bulbs. In the kidney and heart, a reduction of 60-80% ID/g was observed, while an increase was observed in the adrenals requiring 10 mg/kg for significant displacement. Flumazenil had no effect on uptake in peripheral organs and brain. Metabolite analysis indicated >90% of the radioactivity in the above tissues was intact [¹²³I]-CLINDE.

Conclusion

[¹²³I]-CLINDE displays high and selective uptake for the PBBS and warrants further development as a probe for imaging PBBS using single photon emission computed tomography (SPECT).

McEnery MW, Snowman AM, Trifiletti RR, Snyder SH. Isolation of the mitochondrial benzodiazepine receptor: association with the voltage-dependent anion channel and the adenine nucleotide carrier. Proc Nat Acad Sci USA 1992;89:3170-4.CrossRefPubMedPubMedCentral

Olson JM, Ciliax BJ, Mancini WR, Young AB. Presence of peripheral-type benzodiazepine binding sites on human erythrocyte membranes. Eur J Pharmacol 1988;152:47-53.CrossRefPubMed

Oke BO, Suarez-Quian CA, Riond J, Ferrara P, Papadopoulos V. Cell surface localization of the peripheral-type benzodiazepine receptor (PBR) in adrenal cortex. Mol Cell Endocrinoly 1992;87:R1-6.CrossRef

Papadopoulos V, Amri H, Li H, Yao Z, Brown RC, Vidic B, et al. Structure, function and regulation of the mitochondrial peripheral-type benzodiazepine receptor. Therapie 2001;56:549-56.PubMed

Benavides J, Quarteronet D, Imbault F, Malgouris C, Uzan A, Renault C, et al. Labelling of “peripheral-type” benzodiazepine binding sites in the rat brain by using [³H]PK 11195, an isoquinoline carboxamide derivative: kinetic studies and autoradiographic localization. J Neurochem 1983;41:1744-50.CrossRefPubMed

Funakoshi T, Chaki S, Okuyama S, Okubo T, Nakazato A, Nagamine M, et al. In vivo receptor labeling of peripheral benzodiazepine receptor by ex vivo binding of [³H]-PK11195. Res Commun Mol Pathol Pharmacol 1999;105:35-41.PubMed

Brown RC, Papadopoulos V. Role of the peripheral-type benzodiazepine receptor in adrenal and brain steroidogenesis. International Review of Neurobiology 2001;46:117-43.CrossRefPubMed

Li H, Yao Z, Degenhardt B, Teper G, Papadopoulos V. Cholesterol binding at the cholesterol recognition/interaction amino acid consensus (CRAC) of the peripheral-type benzodiazepine receptor and inhibition of steroidogenesis by an HIV TAT-CRAC peptide. Proc Natl Acad Sci USA 2001;98:1267-72.CrossRefPubMedPubMedCentral

Culty M, Luo L, Yao ZX, Chen H, Papadopoulos V, Zirkin BR. Cholesterol transport, peripheral benzodiazepine receptor, and steroidogenesis in aging Leydig cells. J Andrology 2002;23:439-47.

10.

Bono F, Lamarche I, Prabonnaud V, Le Fur G, Herbert JM. Peripheral benzodiazepine receptor agonists exhibit potent antiapoptotic activities. Biochem Biophys Res Commun 1999;265:457-61.CrossRefPubMed

11.

Banati RB. Visualising Microglial Activation in Vivo. Glia 2002;40:206-17.CrossRefPubMed

12.

Mattner F, Katsifis A, Staykova M, Ballantyne P, Willenborg DO. Evaluation of a radiolabelled peripheral benzodiazepine receptor ligand in the central nervous system inflammation of experimental autoimmune encephalomyelitis: a possible probe for imaging multiple sclerosis. Eur J Nuc Med Mol Imaging 2005;32:557-63.CrossRef

13.

Junck L, Olson JM, Ciliax BJ, Koeppe RA, Watkins GL, Jewett DM, et al. PET imaging of human gliomas with ligands for the peripheral benzodiazepine binding site. Ann Neurol 1989;26:752-58.CrossRefPubMed

14.

Hardwick M, Fertikh D, Culty M, Li H, Vidic B, Papadopoulos V. Peripheral-type benzodiazepine receptor (PBR) in human breast cancer: correlation of breast cancer cell aggressive phenotype with PBR expression, nuclear localization, and PBR-mediated cell proliferation and nuclear transport of cholesterol. Cancer Res 1999;59:831-42.PubMed

15.

Katz Y, Eitan A, Amiri Z, Gavish M. Dramatic increase in peripheral benzodiazepine binding sites in human colonic adenocarcinoma as compared to normal colon. Eur J Pharmacol 1988;148:483-4.CrossRefPubMed

16.

Katz Y, Ben-Baruch G, Kloog Y, Menczer J, Gavish M. Increased density of peripheral benzodiazepine-binding sites in ovarian carcinomas as compared with benign ovarian tumours and normal ovaries. Clin Sci (Lond) 1990;78:155-8.CrossRef

17.

Batra S, Alenfall J. Characterization of peripheral benzodiazepine receptors in rat prostatic adenocarcinoma. Prostate (N. Y.) 1994;24:269-78.CrossRef

18.

Langer SZ, Arbilla S, Benavides J, Scatton B. Zolpidem and alpidem: two imidazopyridines with selectivity for w-1 and w-3-receptor subtypes. Advances in biochemistry and psychopharmacology. 46 (GABA benzodiazepine receptor subtypes). New York: Raven; 1990. pp. 61-72.

19.

Trapani G, Franco M, Ricciardi L, Latrofa A, Genchi G, Sanna E, et al. Synthesis and binding affinity of 2-phenylimidazo[1,2-a]pyridine derivatives for both central and peripheral benzodiazepine receptors. A new series of high-affinity and selective ligands for the peripheral type. Jf Med Chem 1997;40:3109-18.CrossRef

20.

Katsifis A, Mattner F, Mardon K, Najdosvski L, Dikic B, Kassiou M. Synthesis and evaluation of [¹²³I]Iodo-imidazo[1,2-b]pyridine as a potential tracer for the study of mitochondria benzodiazepine receptor using SPECT. J Labelled Compd Radiopharm 1997;40:620-2.

21.

Katsifis A, Mattner F, Mardon K, Papazian V, Dikic B. Synthesis and evaluation of [¹²³I]Imidazo[1,2-a]pyridines as potential probes for the study of the peripheral benzodiazepine receptors using SPECT. J Nucl Med 1998;39(5):50.

22.

Katsifis A, Mattner F, Dikic B, Papazian V. Synthesis of substituted [¹²³I]imidazo[1,2-a]pyridines as potential probes for the study of the peripheral benzodiazepine receptors using SPECT. Radiochim Acta 2000;88:229-232.CrossRef

23.

Katsifis A, Mattner F, Zhang Z, Dikic B, Papazian V. Synthesis of [¹²³I]N’,N′-dimethyl-6-methyl-(4′-iodophenyl)imidazo[1,2-a]pyridine-3-acetamide for the study of peripheral benzodiazepine receptors using SPECT. J Labelled Compd Radiopharm 2000;43:385-94.CrossRef

24.

Mardon K, Katsifis A, Mattner F, Dikic B, Donald A, Chapman J. Synthesis and pharmacological evaluation of [¹²³I]Imidazo[1,2-a]pyridines as potential probes for the study of the peripheral benzodiazepine receptors using SPECT. Eur J Nucl Med 1998;25(8):890.

25.

Davies LP, Barlin GB, Selley ML. New imidazo[1,2-b]pyridazine ligands for peripheral-type benzodiazepine receptors on mitochondria and monocytes. Life Sci 1995;57:381-6.CrossRef

26.

Katsifis A, Najdosvski L, Mardon K, Mattner F, Barlin B, Davies L, et al. Synthesis and pharmacological evaluation of an [¹²³I]Iodo-imidazo[1,2-b]pyridazine as a potential tracer for the study of mitochondria benzodiazepine receptor using SPECT. J Labelled Compd Radiopharm 1997;40:623-5.

27.

Katsifis A, Barlin G, Mattner F, Dikic B. Synthesis of [¹²³I]iodine labelled imidazo[1,2-b] pyridazines as potential probes for the study of peripheral benzodiazepine receptors using SPECT. Radiochim Acta 2004;92:305-10.CrossRef

28.

Romeo E, Auta J, Kozikowski AP, Ma D, Papadopoulos V, Puia G, et al. 2-Aryl-3-indoleacetamides (FGIN-1): a new class of potent and specific ligands for the mitochondrial DBI receptor (MDR). J Pharmacol Exp Ther 1992;262:971-8.PubMed

29.

Pike VW, Halldin C, Crouzel C, Barre L, Nutt DJ, Osman S, et al. Radioligands for PET studies of central benzodiazepine receptors and PK (peripheral benzodiazepine) binding sites-current status. Nuc Med Biol 1993;20:503-25.CrossRef

30.

Versijpt J, Dumont F, Vanlaere KJ, Decoo D, Santens P, Audenaert K, et al. Assessment of neuroinflammation and microglial activation in Alzheimer’s disease with radiolabelled PK11195 and single photon emission computed tomography - A Pilot Study. Eur Neurol 2003;50:39-47.CrossRefPubMed

31.

Cagnin A, Myers R, Gunn RN, Lawrence AD, Stevens T, Kreutzberg GW, et al. In vivo visualization of activated glia by [¹¹C] (R)-PK11195-PET following herpes encephalitis reveals projected neuronal damage beyond the primary focal lesion. Brain 2001;124:2014-27.CrossRefPubMed

32.

Banati RB, Newcombe J, Gunn RN, Cagnin A, Turkheimer F, Heppner F, et al. The peripheral benzodiazepine binding site in the brain in multiple sclerosis—quantitative in vivo imaging of microglia as a measure of disease activity. Brain 2000;123:2321-37.CrossRefPubMed

33.

Vowinckel E, Reutens D, Becher B, Verge G, Evans A, Owens T, et al. PK11195 binding to the peripheral benzodiazepine receptor as a marker of microglia activation in multiple sclerosis and experimental autoimmune encephalomyelitis. J Neurosci Res 1997;50:345-53.CrossRefPubMed

34.

Gerhard A, Banati RB, Goerres GB, Cagnin A, Myers R, Gunn RN, et al. [¹¹C](R)-PK11195 PET imaging of microglial activation in multiple system atrophy. Neurol 2003;61:686-9.CrossRef

35.

Charbonneau P, Syrota A, Crouzel C, Valois JM, Prenant C, Crouzel M. Peripheral-type benzodiazepine receptors in the living heart characterized by positron emission tomography. Circulation 1986;73:476-83.CrossRefPubMed

36.

Dumont F, Waterhouse RN, Montoya JA, Mattner F, Katsifis A, Kegeles LS, et al. Synthesis and in vivo evaluation of [¹¹C]zolpidem, an imidazopyridine with agonist properties at central benzodiazepine receptors. Nuc Med Biol 2003;30:435-9.CrossRef

37.

Fonia O, Weizman R, Zisman E, Ashkenazi R, Gavish M. Down-regulation of hepatic peripheral-type benzodiazepine receptors caused by acute lead intoxication. Eur J Pharmacol 1995;293:335-9.CrossRefPubMed

38.

Awad M, Gavish M. Binding of [³H]Ro 5-4864 and [³H]PK 11195 to cerebral cortex and peripheral tissues of various species: species differences and heterogeneity in peripheral benzodiazepine binding sites. J Neurochem 1987;49:1407-14.CrossRefPubMed

39.

Trapani G, Franco M, Latrofa A, Ricciardi L, Carotti A, Serra M, et al. Novel 2-phenylimidazo[1,2-a]pyridine derivatives as potent and selective ligands for peripheral benzodiazepine receptors: synthesis, binding affinity, and in vivo studies. J Med Chem 1999;42:3934-41.CrossRefPubMed

40.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.PubMed

41.

Scatchard G. The attractions of proteins for small molecules and ions. Ann N YAcad Sci 1949;51:660-72.CrossRef

42.

Hill AW. The possible effects of the aggregation of the molecules of haemoglobin on its dissociation curves. J Physiol 1910;40:4-7.

43.

Benavides J, Malgouris C, Imbault F, Begassat F, Uzan A, Renault C, et al. “Peripheral type” benzodiazepine binding sites in rat adrenals: binding studies with PK 11195 and autoradiographic localization. Arch Int Pharmacodyn Ther 1983;266:38-49.PubMed

44.

Katsifis A, Mardon K, Mcphee M, Mattner F, Dikic B, Ridley D. Synthesis and evaluation of [I-123]labelled analogues of the partial inverse agonist Ro 15-4513 for the study of diazepam-insensitive benzodiazepine receptors. Nuc Med Biol 1999;26(6):641-9.CrossRef

45.

Avital A, Richter-Levin G, Leschiner S, Spanier I, Veenman L, Weizman A, et al. Acute and repeated swim stress effects on peripheral benzodiazepine receptors in the rat hippocampus, adrenal, and kidney. Neuropsychopharmacology 2001;25:669-78.CrossRefPubMed

46.

Chaki S, Funakoshi T, Yoshikawa R, Okuyama S, Okubo T, Nakazato A, et al. Binding characteristics of [H-3]DAA1106, a novel and selective ligand for peripheral benzodiazepine receptors. Eur J Pharmacol 1999;371(2-3):197-204.CrossRefPubMed

47.

Pisu MG, Papi G, Porcu P, Trapani G, Latrofa A, Biggio G, et al. Binding of [H-3]CB 34, a selective ligand for peripheral benzodiazepine receptors, to rat brain membranes. Eur J Pharmacol 2001;432:129-134.CrossRefPubMed

48.

Doble A, Ferris O, Burgevin MC, Menager J, Uzan A, Dubroeucq MC, et al. Photoaffinity labeling of peripheral-type benzodiazepine-binding sites. Mol Pharmacol 1987;31:42-9.PubMed

49.

Anholt RR, De Souza EB, Oster-Granite ML, Snyder SH. Peripheral-type benzodiazepine receptors: autoradiographic localization in whole-body sections of neonatal rats. J Pharmacol Exp Ther 1985;233:517-26.PubMed

50.

Zisterer DM, Williams DC. Peripheral-type benzodiazepine receptors. Gen Pharmacol 1997;29:305-14.CrossRefPubMed

51.

Gehlert DR, Stephenson DT, Schober DA, Rash K, Clemens JA. Increased expression of peripheral benzodiazepine receptors in the facial nucleus following motor neuron axotomy. Neurochem Int 1997;31:705-13.CrossRefPubMed

52.

Gildersleeve DL, Van Dort ME, Johnson JW, Sherman PS, Wieland DM. Synthesis and evaluation of -iodo-PK11195 for mapping peripheral-type benzodiazepine receptors (omega 3) in heart. Nuc Med Biol 1996;23:23-8.CrossRef

53.

Zhang MR, Maeda J, Furutsuka K, Yoshida Y, Ogawa M, Suhara T, et al. [F-18]FMDAA1106 and [F-18]FEDAA1106: Two positron-emitter labeled ligands for peripheral benzodiazepine receptor (PBR). Bioorg Med Chem Lett 2003;13:201-4.CrossRefPubMed

54.

Zhang MR, Ogawa M, Maeda J, Ito T, Noguchi J, Kumata K, et al. [2-c-11]isopropyl-, [1-c-11]ethyl-, and [c-11]methyl-labeled phenoxyphenyl acetamide derivatives as positron emission tomography ligands for the peripheral benzodiazepine receptor: radiosynthesis, uptake, and in vivo binding in brain. J Med Chem 2006;49:2735-42.CrossRefPubMed

55.

Katsifis A, Mattner F, Mana P, Linares D, Ballantyne P, Berghofer P, et al. Evaluation of CLINDE, a radiolabelled peripheral benzodiazepine receptor ligand in a murine model of neuro-inflammation: potential probe for assessing neurodegeneration. J Labelled Compd Radiopharm 2005;48:S88.

Title: Pharmacological evaluation of [123I]-CLINDE: a radioiodinated imidazopyridine-3-acetamide for the study of peripheral benzodiazepine binding sites (PBBS)
Authors: Filomena Mattner
Karine Mardon
Andrew Katsifis
Publication date: 01-04-2008
Publisher: Springer Berlin Heidelberg
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 4/2008
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-007-0645-5

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Pharmacological evaluation of [¹²³I]-CLINDE: a radioiodinated imidazopyridine-3-acetamide for the study of peripheral benzodiazepine binding sites (PBBS)

Abstract

Purpose

Materials and methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Materials and methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 4/2008

The many roads to infection imaging

Whole body [11C]-dihydrotetrabenazine imaging of baboons: biodistribution and human radiation dosimetry estimates

Combined use of zoledronic acid and 153Sm-EDTMP in hormone-refractory prostate cancer patients with bone metastases

GMP—three letters with many interpretations

Radiation dosimetry and biodistribution of 11C-ABP688 measured in healthy volunteers

“Wispino” protocol for 18FDG-PET scanning of myocardium