title: A 99mTc-Labeled Ligand of Carbonic Anhydrase IX Selectively Targets Renal Cell Carcinoma In Vivo.
abstract: Small organic ligands, selective for tumor-associated antigens, are increasingly being considered as alternatives to monoclonal antibodies for the targeted delivery of diagnostic and therapeutic payloads such as radionuclides and drugs into neoplastic masses. We have previously described a novel acetazolamide derivative, a carbonic anhydrase ligand with high affinity for the tumor-associated isoform IX (CAIX), which can transport highly potent cytotoxic drugs into CAIX-expressing solid tumors. The aim of the present study was to quantitatively investigate the biodistribution properties of said ligand and understand whether acetazolamide conjugates merit further development as drug carriers and radioimaging agents.
  
  
  
  The conjugate described in this study, consisting of a derivative of acetazolamide, a spacer, and a peptidic (99m)Tc chelator, was labeled using sodium pertechnetate under reducing conditions and injected intravenously into CAIX-expressing SKRC-52 xenograft-bearing mice. Animals were sacrificed, and organ uptake as percentage injected activity of radiolabeled ligand per gram of tissues (%IA/g) was evaluated between 10 min and 24 h. Additionally, postmortem imaging by SPECT was performed.
  
  
  
  The acetazolamide conjugate described in this study could be labeled to high radiochemical purity (>95%, 2.2-4.5 MBq/nmol). Analysis of organ uptake at various time points revealed that the ligand displayed a maximal tumor accumulation 3 h after intravenous injection (22 %IA/g), with an excellent tumor-to-blood ratio of 70:1 at the same time point. The ligand accumulation in the tumor was more efficient than in any other organ, but a residual uptake in the kidney, lung, and stomach (9, 16, and 10 %IA/g, respectively) was also observed, in line with patterns of carbonic anhydrase isoform expression in those tissues. Interestingly, tumor-to-organ ratios improved on administration of higher doses of radiolabeled ligand, suggesting that certain binding sites in normal organs can be saturated in vivo.
  
  
  
  The (99m)Tc-labeled acetazolamide conjugate exhibits high tumor uptake and favorable tumor-to-kidney ratios of up to 3 that may allow imaging of tumors in the kidney and distant sites at earlier time points than commonly possible with antibody-based products. These data suggest that the described molecule merit further development as a radioimaging agent for CAIX-expressing renal cell carcinoma.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zurich, Switzerland.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Krall
authors.affiliation: Philochem AG, Otelfingen (ZH), Switzerland; and.
authors.forename: Francesca
authors.initials: F
authors.lastname: Pretto
authors.affiliation: Philochem AG, Otelfingen (ZH), Switzerland; and.
authors.forename: Martin
authors.initials: M
authors.lastname: Mattarella
authors.affiliation: Research Department Biology and Chemistry, Paul Scherrer Institut, Center for Radiopharmaceutical Sciences, Villigen-PSI, Switzerland.
authors.forename: Cristina
authors.initials: C
authors.lastname: Müller
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zurich, Switzerland neri@pharma.ethz.ch.
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
mesh_terms: D000086:Acetazolamide / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D000818:Animals; D000071231:Carbonic Anhydrase IX / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000000981:diagnostic imaging / Q000378:metabolism*; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007553:Isotope Labeling; D007680:Kidney Neoplasms / Q000000981:diagnostic imaging / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D013667:Technetium / Q000737:chemistry*; D014018:Tissue Distribution; D015899:Tomography, Emission-Computed, Single-Photon
chemical_list: D008024:Ligands; D013667:Technetium; D000071231:Carbonic Anhydrase IX; D000086:Acetazolamide
keywords: carbonic anhydrase IX; imaging agent; kidney cancer; tumor targeting