title: Preclinical evaluation of the novel hypoxic marker 99mTc-HL91 (Prognox) in murine and xenograft systems in vivo.
abstract: OBJECTIVE
  The 99mTc-labelled amine oxime 99mTc-HL91 (Prognox) is under investigation as a potential noninvasive clinical marker of tumour hypoxia whose uptake can be monitored by gamma camera imaging. The aim was to assess its retention in 3 tumours under control and enhanced oxygenation conditions.
  
  
  METHODS
  The SaF murine sarcoma, grown subcutaneously in CBA mice, and human colon carcinoma HT29 and lung adenocarcinoma A549, grown as xenografts in SCID mice, were used at 6-8 mm diameter. Oxygenation status was enhanced by giving 500 mg/kg nicotinamide i.p. and breathing carbogen until the point of assay. Oxygenation/hypoxia was measured using the Eppendorf pO2 histograph (KIMOC 6650) with at least 5 tracks and at least 70 values, and expressing pO2 values as % < 2.5 mmHg. 99mTc-HL91 (0.8 or 8 MBq per mouse) was injected i.v. immediately before nicotinamide or saline, and animals were killed 2 h after injection. Tumour, skin, muscle, and blood samples were counted and isotope retention was expressed as % injected dose per gram. 14C-labelled uncomplexed HL91 was used similarly (0.2-0.4 MBq per mouse) and samples were solubilised and decolourised before counting.
  
  
  RESULTS
  Nicotinamide and carbogen treatment reduced 99mTc-HL91 retention in all tumours to 54%-64% of control; it also reduced the proportion of pO2 values < 2.5 mmHg in all tumours. The mean proportion of pO2 values < 2.5 mmHg correlated very well with the mean ratio of tumour to blood retention at 2 h for all tumours, both unperturbed and oxygen-enhanced (r = 0.996, p < 0.001). Retention of 14C-HL91 in SaF tumour was unchanged by nicotinamide and carbogen, confirming that 99mTc complexation of the ligand is required for hypoxia specificity.
  
  
  CONCLUSIONS
  There is excellent correlation between 99mTc-HL91 retention and hypoxia, as measured by the Eppendorf histograph, over the range of 50%-90% of values < 2.5 mmHg in 3 different tumour models, including 2 human xenografts. 99mTc complexation of the ligand is required for hypoxia specificity. 99mTc-HL91 (Prognox) shows good potential as a clinical marker for hypoxia and warrants further development.
authors.affiliation: Tumour Microcirculation Group, Gray Laboratory Cancer Research Trust, Mount Vernon Hospital, Northwood, Middlesex, UK.
authors.forename: D J
authors.initials: DJ
authors.lastname: Honess
authors.forename: S A
authors.initials: SA
authors.lastname: Hill
authors.forename: D R
authors.initials: DR
authors.lastname: Collingridge
authors.forename: B
authors.initials: B
authors.lastname: Edwards
authors.forename: G
authors.initials: G
authors.lastname: Brauers
authors.forename: N A
authors.initials: NA
authors.lastname: Powell
authors.forename: D J
authors.initials: DJ
authors.lastname: Chaplin
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism; D000818:Animals; D002245:Carbon Dioxide / Q000008:administration & dosage; D002277:Carcinoma / Q000378:metabolism; D015687:Cell Hypoxia; D003110:Colonic Neoplasms / Q000378:metabolism; D003287:Contrast Media / Q000493:pharmacokinetics*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism; D051379:Mice; D008808:Mice, Inbred CBA; D016513:Mice, SCID; D009536:Niacinamide / Q000008:administration & dosage; D015609:Organotechnetium Compounds / Q000493:pharmacokinetics*; D010091:Oximes / Q000493:pharmacokinetics*; D010100:Oxygen / Q000008:administration & dosage; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics*; D012509:Sarcoma / Q000378:metabolism
chemical_list: D003287:Contrast Media; D015609:Organotechnetium Compounds; D010091:Oximes; D019275:Radiopharmaceuticals; C105527:technetium Tc 99m 4,9-diaza-3,3,10,10-tetramethyldodecan-2,11-dione dioxime; D002245:Carbon Dioxide; D009536:Niacinamide; C011700:carbogen; D010100:Oxygen