PSMA ligands for radionuclide imaging and therapy of prostate cancer: Clinical status

Theranostics

Volumn 5, Issue 12, 2015, Pages 1388-1401

  Lütje, Susanne ^a,b   Heskamp, Sandra ^b   Cornelissen, Alexander S ^b   Poeppel, Thorsten D ^a   van den Broek, Sebastiaan A M W ^c   Rosenbaum Krumme, Sandra ^a   Bockisch, Andreas ^a   Gotthardt, Martin ^b   Rijpkema, Mark ^b   Boerman, Otto C ^b  

^a UNIVERSITY HOSPITAL ESSEN   (Germany)

^b RADBOUD UNIVERSITY MEDICAL CENTER   (Netherlands)

^c FutureChemistry Holding BV   (Netherlands)

Author keywords

PET; Prostate cancer; PSMA; Radionuclide imaging; Theranostics

Indexed keywords

2 [3 [1 CARBOXY 5 [(6 FLUOROPYRIDINE 3 CARBONYL)AMINO]PENTYL]UREIDO]PENTANEDIOIC ACID F 18; 2 [3 [1 CARBOXY 5 [3 (4 IODOPHENYL)UREIDO]PENTYL]UREIDO]PENTANEDIOIC ACID I 123; 2 [3 [1 CARBOXY 5 [3 (4 IODOPHENYL)UREIDO]PENTYL]UREIDO]PENTANEDIOIC ACID I 131; CAPROMAB PENDETIDE IN 111; IOFOLASTAT I 123; MIP 1405 TC 99M; MONOCLONAL ANTIBODY J591 LU 177; N [N (1,3 DICARBOXYPROPYL)CARBAMOYL] 4 FLUOROBENZYL CYSTEINE F 18; PROSTATE SPECIFIC ANTIGEN; PROSTATE SPECIFIC MEMBRANE ANTIGEN; RADIOPHARMACEUTICAL AGENT; TETRAXETAN GA 68; TROFOLASTAT TC 99M; UNCLASSIFIED DRUG; GLUTAMATE CARBOXYPEPTIDASE II; GLUTAMATE CARBOXYPEPTIDASE II, HUMAN; LIGAND; MEMBRANE ANTIGEN; PROTEIN BINDING;

ANTIGEN DETECTION; ANTIGEN EXPRESSION; BINDING AFFINITY; BLOOD TOXICITY; BONE SCINTISCANNING; CANCER GRADING; CANCER MORTALITY; CANCER STAGING; CASTRATION RESISTANT PROSTATE CANCER; COMPUTER ASSISTED EMISSION TOMOGRAPHY; COMPUTER ASSISTED TOMOGRAPHY; DIAGNOSTIC IMAGING; DRUG CLEARANCE; DRUG DISTRIBUTION; DRUG HALF LIFE; DRUG UPTAKE; GLEASON SCORE; HUMAN; IMAGE ANALYSIS; ISOTOPE LABELING; LIGAND BINDING; MORBIDITY; NONHUMAN; NUCLEAR MAGNETIC RESONANCE IMAGING; OUTCOME ASSESSMENT; PROSTATE CANCER; REVIEW; SCINTISCANNING; SINGLE DRUG DOSE; SINGLE PHOTON EMISSION COMPUTER TOMOGRAPHY; THERANOSTIC NANOMEDICINE; UPREGULATION; MALE; METABOLISM; PROCEDURES; PROSTATIC NEOPLASMS;

ANTIGENS, SURFACE; GLUTAMATE CARBOXYPEPTIDASE II; HUMANS; LIGANDS; MALE; PROSTATIC NEOPLASMS; PROTEIN BINDING; RADIONUCLIDE IMAGING; RADIOTHERAPY;

EID: 84954239466     PISSN: None     EISSN: 18387640     Source Type: Journal    
DOI: 10.7150/thno.13348     Document Type: Review

References (78)

1. Jana S, Blaufox MD. Nuclear medicine studies of the prostate, testes, and bladder. Semin Nucl Med. 2006; 36: 51-72.
2. Gambhir SS, Czernin J, Schwimmer J, Silverman DH, Coleman RE, Phelps ME. A tabulated summary of the FDG PET literature. J Nucl Med. 2001; 42: 1S-93S.
3. Bouchelouche K, Tagawa ST, Goldsmith SJ, Turkbey B, Capala J, Choyke P. PET/CT Imaging and Radioimmunotherapy of Prostate Cancer. Semin Nucl Med. 2011; 41: 29-44.
4. Ackerstaff E, Glunde K, Bhujwalla ZM. Choline phospholipid metabolism: a target in cancer cells? J Cell Bioochem. 2003; 90: 525-33.
5. Glunde K, Ackerstaff E, Mori N, Jacobs MA, Bhujwalla ZM. Choline phospholipid metabolism in cancer: consequences for molecular pharmaceutical interventions. Mol Pharm. 2006; 3: 496-506.
6. Kitajima K, Murphy RC, Nathan MA. Choline PET/CT for imaging prostate cancer: an update. Ann Nucl Med. 2013; 27: 581-91.
7. Umbehr MH, Muntener M, Hany T, Sulser T, Bachmann LM. The role of 11C-choline and 18F-fluorocholine positron emission tomography (PET) and PET/CT in prostate cancer: a systematic review and meta-analysis. Eur Urol. 2013; 64: 106-17.
8. Krause BJ, Souvatzoglou M, Treiber U. Imaging of prostate cancer with PET/CT and radioactively labeled choline derivates. Urol Oncol. 2013; 31: 427-35.
9. Bauman G, Belhocine T, Kovacs M, Ward A, Beheshti M, Rachinsky I. 18F-fluorocholine for prostate cancer imaging: a systematic review of the literature. Prostate cancer Prostatic Dis. 2012; 15: 45-55.
10. Castellucci P, Ceci F, Graziani T, Schiavina R, Brunocilla E, Mazzarotto R, et al. Early biochemical relapse after radical prostatectomy: which prostate cancer patients may benefit from a restaging 11C-Choline PET/CT scan before salvage radiation therapy? J Nucl Med. 2014; 55: 1424-9.
11. Lutje S, Boerman OC, van Rij CM, Sedelaar M, Helfrich W, Oyen WJ, et al. Prospects in radionuclide imaging of prostate cancer. Prostate. 2012; 72: 1262-72.
12. Israeli RS, Powell CT, Fair WR, Heston WD. Molecular cloning of a complementary DNA encoding a prostate-specific membrane antigen. Cancer Res. 1993; 53: 227-30.
13. Zhou J, Neale JH, Pomper MG, Kozikowski AP. NAAG peptidase inhibitors and their potential for diagnosis and therapy. Nat Rev Drug Discov. 2005; 4: 1015-26.
14. Silver DA, Pellicer I, Fair WR, Heston WD, Cordon-Cardo C. Prostate-specific membrane antigen expression in normal and malignant human tissues. Clin Cancer Res. 1997; 3: 81-5.
15. Conway RE, Petrovic N, Li Z, Heston W, Wu D, Shapiro LH. Prostate-specific membrane antigen regulates angiogenesis by modulating integrin signal transduction. Mol Cell Biol. 2006; 26: 5310-24.
16. Minner S, Wittmer C, Graefen M, Salomon G, Steuber T, Haese A, et al. High level PSMA expression is associated with early PSA recurrence in surgically treated prostate cancer. Prostate. 2011; 71: 281-8.
17. Rybalov M, Ananias HJ, Hoving HD, van der Poel HG, Rosati S, de Jong IJ. PSMA, EpCAM, VEGF and GRPR as imaging targets in locally recurrent prostate cancer after radiotherapy. Int J Mol Sci. 2014; 15: 6046-61.
18. Ananias HJ, van den Heuvel MC, Helfrich W, de Jong IJ. Expression of the gastrin-releasing peptide receptor, the prostate stem cell antigen and the prostate-specific membrane antigen in lymph node and bone metastases of prostate cancer. Prostate. 2009; 69: 1101-8.
19. Kawakami M, Nakayama J. Enhanced expression of prostate-specific membrane antigen gene in prostate cancer as revealed by in situ hybridization. Cancer Res. 1997; 57: 2321-4.
20. Bostwick DG, Pacelli A, Blute M, Roche P, Murphy GP. Prostate specific membrane antigen expression in prostatic intraepithelial neoplasia and adenocarcinoma: a study of 184 cases. Cancer. 1998; 82: 2256-61.
21. Graham K, Lesche R, Gromov AV, Bohnke N, Schafer M, Hassfeld J, et al. Radiofluorinated derivatives of 2-(phosphonomethyl)pentanedioic acid as inhibitors of prostate specific membrane antigen (PSMA) for the imaging of prostate cancer. J Med Chem. 2012; 55: 9510-20.
22. Troyer JK, Beckett ML, Wright GL, Jr. Detection and characterization of the prostate-specific membrane antigen (PSMA) in tissue extracts and body fluids. Int J Cancer. 1995; 62: 552-8.
23. Wright GL, Jr., Grob BM, Haley C, Grossman K, Newhall K, Petrylak D, et al. Upregulation of prostate-specific membrane antigen after androgen-deprivation therapy. Urology. 1996; 48: 326-34.
24. Ross JS, Sheehan CE, Fisher HA, Kaufman RP, Jr., Kaur P, Gray K, et al. Correlation of primary tumor prostate-specific membrane antigen expression with disease recurrence in prostate cancer. Clin Cancer Res. 2003; 9: 6357-62.
25. Horoszewicz JS, Kawinski E, Murphy GP. Monoclonal antibodies to a new antigenic marker in epithelial prostatic cells and serum of prostatic cancer patients. Anticancer Res. 1987; 7: 927-35.
26. Franc BL, Cho SY, Rosenthal SA, Cui Y, Tsui B, Vandewalker KM, et al. Detection and localization of carcinoma within the prostate using high resolution transrectal gamma imaging (TRGI) of monoclonal antibody directed at prostate specific membrane antigen (PSMA)-proof of concept and initial imaging results. Eur J Radiol. 2013; 82: 1877-84.
27. Ponsky LE, Cherullo EE, Starkey R, Nelson D, Neumann D, Zippe CD. Evaluation of preoperative ProstaScint scans in the prediction of nodal disease. Prostate cancer Prostatic Dis. 2002; 5: 132-5.
28. Liu H, Rajasekaran AK, Moy P, Xia Y, Kim S, Navarro V, et al. Constitutive and antibody-induced internalization of prostate-specific membrane antigen. Cancer Res. 1998; 58: 4055-60.
29. Bander NH, Trabulsi EJ, Kostakoglu L, Yao D, Vallabhajosula S, Smith-Jones P, et al. Targeting metastatic prostate cancer with radiolabeled monoclonal antibody J591 to the extracellular domain of prostate specific membrane antigen. J Urol. 2003; 170: 1717-21.
30. Tagawa ST, Milowsky MI, Morris M, Vallabhajosula S, Christos P, Akhtar NH, et al. Phase II study of Lutetium-177-labeled anti-prostate-specific membrane antigen monoclonal antibody J591 for metastatic castration-resistant prostate cancer. Clinical Cancer Res. 2013; 19: 5182-91.
31. Tsukamoto T, Wozniak KM, Slusher BS. Progress in the discovery and development of glutamate carboxypeptidase II inhibitors. Drug Discov Today. 2007; 12: 767-76.
32. Foss CA, Mease RC, Fan H, Wang Y, Ravert HT, Dannals RF, et al. Radiolabeled small-molecule ligands for prostate-specific membrane antigen: in vivo imaging in experimental models of prostate cancer. Clin Cancer Res. 2005; 11: 4022-8.
33. Mease RC, Dusich CL, Foss CA, Ravert HT, Dannals RF, Seidel J, et al. N-[N-[(S)-1,3-Dicarboxypropyl]carbamoyl]-4-[18F]fluorobenzyl-L-cysteine, [18F]DCFBC: a new imaging probe for prostate cancer. Clin Cancer Res. 2008; 14: 3036-43.
34. Maresca KP, Hillier SM, Femia FJ, Keith D, Barone C, Joyal JL, et al. A series of halogenated heterodimeric inhibitors of prostate specific membrane antigen (PSMA) as radiolabeled probes for targeting prostate cancer. J Med Chem. 2009; 52: 347-57.
35. Banerjee SR, Foss CA, Castanares M, Mease RC, Byun Y, Fox JJ, et al. Synthesis and evaluation of technetium-99m-and rhenium-labeled inhibitors of the prostate-specific membrane antigen (PSMA). J Med Chem. 2008; 51: 4504-17.
36. Hillier SM, Maresca KP, Lu G, Merkin RD, Marquis JC, Zimmerman CN, et al. 99mTc-labeled small-molecule inhibitors of prostate-specific membrane antigen for molecular imaging of prostate cancer. J Nucl Med. 2013; 54: 1369-76.
37. Banerjee SR, Pullambhatla M, Byun Y, Nimmagadda S, Green G, Fox JJ, et al. 68Ga-labeled inhibitors of prostate-specific membrane antigen (PSMA) for imaging prostate cancer. J Med Chem. 2010; 53: 5333-41.
38. Schafer M, Bauder-Wust U, Leotta K, Zoller F, Mier W, Haberkorn U, et al. A dimerized urea-based inhibitor of the prostate-specific membrane antigen for 68Ga-PET imaging of prostate cancer. EJNMMI Res. 2012; 2: 23.
39. Chen Y, Pullambhatla M, Banerjee SR, Byun Y, Stathis M, Rojas C, et al. Synthesis and biological evaluation of low molecular weight fluorescent imaging agents for the prostate-specific membrane antigen. Bioconjug Chem. 2012; 23: 2377-85.
40. Mesters JR, Barinka C, Li W, Tsukamoto T, Majer P, Slusher BS, et al. Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. EMBO J. 2006; 25: 1375-84.
41. Huang SS, Wang X, Zhang Y, Doke A, DiFilippo FP, Heston WD. Improving the biodistribution of PSMA-targeting tracers with a highly negatively charged linker. Prostate. 2014; 74: 702-13.
42. Benesova M, Schafer M, Bauder-Wust U, Afshar-Oromieh A, Kratochwil C, Mier W, et al. Preclinical evaluation of a tailor-made DOTA-conjugated PSMA inhibitor with optimized linker moiety for imaging and endoradiotherapy of prostate cancer. J Nucl Med. 2015.
43. Barrett JA, Coleman RE, Goldsmith SJ, Vallabhajosula S, Petry NA, Cho S, et al. First-in-man evaluation of 2 high-affinity PSMA-avid small molecules for imaging prostate cancer. J Nucl Med. 2013; 54: 380-7.
44. Vallabhajosula S, Nikolopoulou A, Babich JW, Osborne JR, Tagawa ST, Lipai I, et al. 99mTc-labeled small-molecule inhibitors of prostate-specific membrane antigen: pharmacokinetics and biodistribution studies in healthy subjects and patients with metastatic prostate cancer. J Nucl Med. 2014; 55: 1791-8.
45. Cho SY, Gage KL, Mease RC, Senthamizhchelvan S, Holt DP, Jeffrey-Kwanisai A, et al. Biodistribution, tumor detection, and radiation dosimetry of 18F-DCFBC, a low-molecular-weight inhibitor of prostate-specific membrane antigen, in patients with metastatic prostate cancer. J Nucl Med. 2012; 53: 1883-91.
46. Zechmann CM, Afshar-Oromieh A, Armor T, Stubbs JB, Mier W, Hadaschik B, et al. Radiation dosimetry and first therapy results with a (124)I/(131)I-labeled small molecule (MIP-1095) targeting PSMA for prostate cancer therapy. Eur J Nucl Med Mol Imaging. 2014; 41: 1280-92.
47. Afshar-Oromieh A, Zechmann CM, Malcher A, Eder M, Eisenhut M, Linhart HG, et al. Comparison of PET imaging with a (68)Ga-labelled PSMA ligand and (18)F-choline-based PET/CT for the diagnosis of recurrent prostate cancer. Eur J Nucl Med Mol Imaging. 2014; 41: 11-20.
48. Afshar-Oromieh A, Malcher A, Eder M, Eisenhut M, Linhart HG, Hadaschik BA, et al. PET imaging with a [68Ga]gallium-labelled PSMA ligand for the diagnosis of prostate cancer: biodistribution in humans and first evaluation of tumour lesions. Eur J Nucl Med Mol Imaging. 2013; 40: 486-95.
49. Afshar-Oromieh A, Haberkorn U, Schlemmer HP, Fenchel M, Eder M, Eisenhut M, et al. Comparison of PET/CT and PET/MRI hybrid systems using a 68Ga-labelled PSMA ligand for the diagnosis of recurrent prostate cancer: initial experience. Eur J Nucl Med Mol Imaging. 2014; 41: 887-97.
50. Hillier SM, Maresca KP, Femia FJ, Marquis JC, Foss CA, Nguyen N, et al. Preclinical evaluation of novel glutamate-urea-lysine analogues that target prostate-specific membrane antigen as molecular imaging pharmaceuticals for prostate cancer. Cancer Res. 2009; 69: 6932-40.
51. Maresca KP, Marquis JC, Hillier SM, Lu G, Femia FJ, Zimmerman CN, et al. Novel polar single amino acid chelates for technetium-99m tricarbonyl-based radiopharmaceuticals with enhanced renal clearance: application to octreotide. Bioconjug Chem. 2010; 21: 1032-42.
52. O'Keefe DS, Bacich DJ, Heston WD. Comparative analysis of prostate-specific membrane antigen (PSMA) versus a prostate-specific membrane antigen-like gene. Prostate. 2004; 58: 200-10.
53. Reubi JC, Maecke HR. Peptide-based probes for cancer imaging. J Nucl Med. 2008; 49: 1735-8.
54. Roesch F, Riss PJ. The renaissance of the (6)(8)Ge/(6)(8)Ga radionuclide generator initiates new developments in (6)(8)Ga radiopharmaceutical chemistry. Curr Top Med Chem. 2010; 10: 1633-68.
55. Eder M, Schafer M, Bauder-Wust U, Hull WE, Wangler C, Mier W, et al. 68Ga-complex lipophilicity and the targeting property of a urea-based PSMA inhibitor for PET imaging. Bioconjug Chem. 2012; 23: 688-97.
56. Morigi JJ, Stricker P, Van Leeuwen P, Tang R, Ho B, Nguyen Q, et al. Prospective Comparison of the detection rate of 18F-Fluoromethylcholine and 68Ga-PSMA-HBED PET/CT in men with prostate cancer with rising PSA post curative treatment, being considered for targeted therapy. J Nucl Med. 2015; 56: 1185-90.
57. Afshar-Oromieh A, Avtzi E, Giesel FL, Holland-Letz T, Linhart HG, Eder M, et al. The diagnostic value of PET/CT imaging with the (68)Ga-labelled PSMA ligand HBED-CC in the diagnosis of recurrent prostate cancer. Eur J Nucl Med Mol Imaging. 2015; 42: 197-209.
58. Eiber M, Maurer T, Souvatzoglou M, Beer AJ, Ruffani A, Haller B, et al. Evaluation of Hybrid 68Ga-PSMA Ligand PET/CT in 248 Patients with Biochemical Recurrence After Radical Prostatectomy. J Nucl Med. 2015; 56: 668-74.
59. Brogsitter C, Zophel K, Kotzerke J. 18F-Choline, 11C-choline and 11C-acetate PET/CT: comparative analysis for imaging prostate cancer patients. Eur J Nucl Med Mol Imaging. 2013; 40 Suppl 1: S18-27.
60. Ceci F, Uprimny C, Nilica B, Geraldo L, Kendler D, Kroiss A, et al. (68)Ga-PSMA PET/CT for restaging recurrent prostate cancer: which factors are associated with PET/CT detection rate? Eur J Nucl Med Mol Imaging. 2015; 42: 1284-94.
61. Krohn T, Verburg FA, Pufe T, Neuhuber W, Vogg A, Heinzel A, et al. [(68)Ga]PSMA-HBED uptake mimicking lymph node metastasis in coeliac ganglia: an important pitfall in clinical practice. Eur J Nucl Med Mol Imaging. 2015; 42: 210-4.
62. Rowe SP, Gage KL, Faraj SF, Macura KJ, Cornish TC, Gonzalez-Roibon N, et al. 18F-DCFBC PET/CT for PSMA-Based Detection and Characterization of Primary Prostate Cancer. J Nucl Med. 2015; 56: 1003-10.
63. Szabo Z, Mena E, Rowe SP, Plyku D, Nidal R, Eisenberger MA, et al. Initial Evaluation of [(18)F]DCFPyL for Prostate-Specific Membrane Antigen (PSMA)-Targeted PET Imaging of Prostate Cancer. Mol Imaging Biol. 2015; 17: 565-74.
64. Chen Y, Pullambhatla M, Foss CA, Byun Y, Nimmagadda S, Senthamizhchelvan S, et al. 2-(3-(1-Carboxy-5-[(6-[18F]fluoro-pyridine-3-carbonyl)-amino]-pentyl)-ureido )-pen tanedioic acid, [18F]DCFPyL, a PSMA-based PET imaging agent for prostate cancer. Clin Cancer Res. 2011; 17: 7645-53.
65. Dietlein M, Kobe C, Kuhnert G, Stockter S, Fischer T, Schomacker K, et al. Comparison of [(18)F]DCFPyL and [ (68)Ga]Ga-PSMA-HBED-CC for PSMA-PET Imaging in Patients with Relapsed Prostate Cancer. Mol Imaging Biol. 2015; 17: 575-84.
66. Bander NH, Milowsky MI, Nanus DM, Kostakoglu L, Vallabhajosula S, Goldsmith SJ. Phase I trial of 177lutetium-labeled J591, a monoclonal antibody to prostate-specific membrane antigen, in patients with androgen-independent prostate cancer. J Clin Oncol. 2005; 23: 4591-601.
67. Kinoshita Y, Kuratsukuri K, Landas S, Imaida K, Rovito PM, Jr., Wang CY, et al. Expression of prostate-specific membrane antigen in normal and malignant human tissues. World J Surg. 2006; 30: 628-36.
68. Kassis AI. Therapeutic radionuclides: biophysical and radiobiologic principles. Semin Nucl Med. 2008; 38: 358-66.
69. Benesova M, Schafer M, Bauder-Wust U, Afshar-Oromieh A, Kratochwil C, Mier W, et al. Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer. J Nucl Med. 2015; 56: 914-20.
70. Kratochwil C, Giesel FL, Eder M, Afshar-Oromieh A, Benesova M, Mier W, et al. [(177)Lu]Lutetium-labelled PSMA ligand-induced remission in a patient with metastatic prostate cancer. Eur J Nucl Med Mol Imaging. 2015; 42: 987-8.
71. Baum RP, Wahl RL. Third Theranostics World Congress on Gallium-68 and PRRT: Abstracts. J Nucl Med. 2015; 56 Suppl 2: 2A-30.
72. Kratochwil C, Giesel FL, Leotta K, Eder M, Hoppe-Tich T, Youssoufian H, et al. PMPA for nephroprotection in PSMA-targeted radionuclide therapy of prostate cancer. J Nucl Med. 2015; 56: 293-8.
73. Weineisen M, Simecek J, Schottelius M, Schwaiger M, Wester HJ. Synthesis and preclinical evaluation of DOTAGA-conjugated PSMA ligands for functional imaging and endoradiotherapy of prostate cancer. EJNMMI Res 2014; 4: 63.
74. Weineisen M, Schottelius M, Simecek J, Baum RP, Yildiz A, Beykan S, et al. 68Ga-and 177Lu-labeled PSMA I&T: Optimization of a PSMA targeted theranostic concept and first proof of concept human studies. J Nucl Med. 2015; 56: 1169-76.
75. Herrmann K, Bluemel C, Weineisen M, Schottelius M, Wester HJ, Czernin J, et al. Biodistribution and radiation dosimetry for a probe targeting prostate-specific membrane antigen for imaging and therapy. J Nucl Med. 2015; 56: 855-61.
76. Neuman BP, Eifler JB, Castanares M, Chowdhury WH, Chen Y, Mease RC, et al. Real-time, near-infrared fluorescence imaging with an optimized dye/light source/camera combination for surgical guidance of prostate cancer. Clin Cancer Res. 2015; 21: 771-80.
77. Maurer T, Weirich G, Schottelius M, Weineisen M, Frisch B, Okur A, et al. Prostate-specific Membrane Antigen-radioguided Surgery for Metastatic Lymph Nodes in Prostate Cancer. Eur Urol. 2015; 68: 530-4.
78. Chen Y, Dhara S, Banerjee SR, Byun Y, Pullambhatla M, Mease RC, et al. A low molecular weight PSMA-based fluorescent imaging agent for cancer. Biochem Biophys Res Commun. 2009; 390: 624-9.