Biological evaluation of complexes of cyclopentadienyl M(CO)3+ (M = Re, 99mTc) with high blood-brain barrier penetration potential as brain cancer agents.

To address the major medical need for effective chemotherapeutics/diagnostics for brain cancer, in this work three cyclopentadienyl M(CO)3+ (M = Re, 99mTc) complexes, which cross the blood-brain barrier (BBB) in high % and are designed to mimic the anticancer agent 2-phenylbenzothiazole, are in vitro and in vivo evaluated for anticancer action. The study includes cytotoxicity and uptake studies in cancer and healthy neuronal cell lines, mechanistic investigation of potential anticancer pathways, and biodistribution studies in mice bearing glioblastoma xenografts. The stable Re complexes exhibit selective uptake and significant antiproliferative effect, particularly against U-251 MG glioblastoma cells, with no significant toxicity in healthy neurons, demonstrating the suitability of this type of complexes to serve as selective therapeutic/imaging agents for brain cancer. Furthermore, they result in the generation of elevated Reactive Oxygen Species (ROS) levels, and lead to significant G2/M arrest followed by apoptosis. Biodistribution studies in U-251 MG xenograft bearing mice with the radioactive 99mTc complex that exhibits the highest BBB penetration, show retention at the tumor-site offering a diagnostic prospect and, in addition, indicating the capability of the Re analogue to accumulate at the tumor site for therapeutic action. Overall, the complexes demonstrate significant anticancer properties that, combined with their high BBB penetration potential, render them strong candidates for further evaluation as brain cancer agents.

Bare and protein-conjugated Fe(3)O(4) ferromagnetic nanoparticles for utilization in magnetically assisted hemodialysis: biocompatibility with human blood cells.

Magnetically assisted hemodialysis is a development of conventional hemodialysis and is based on the circulation of ferromagnetic nanoparticle-targeted binding substance conjugates (FN-TBS Cs) in the bloodstream of the patient and their eventual removal by means of a 'magnetic dialyzer'. Presented here is an in vitro investigation on the biocompatibility of bare Fe(3)O(4) FNs and Fe(3)O(4)-bovine serum albumin Cs with blood cells, namely red blood cells (RBCs), white blood cells (WBCs) and platelets (Plts). Atomic force microscopy (AFM) and optical microscopy (OM) enabled the examination of blood cells at the nanometer and micrometer level, respectively. The observations made on FN- and C-maturated blood samples are contrasted to those obtained on FN- and C-free reference blood samples subjected to exactly the same maturation procedure. Qualitatively, both AFM and OM revealed no changes in the overall shape of RBCs, WBCs and Plts. Incidents where bare FNs or Cs were bound onto the surface of RBCs or internalized by WBCs were very rare. Detailed examination by means of OM proved that impaired coagulation of Plts is not initiated/promoted either by FNs or Cs. Quantitatively, the statistical analysis of the obtained AFM images from RBC surfaces clearly revealed that the mean surface roughness of RBCs maturated with bare FNs or Cs was identical to the one of reference RBCs.

In vitro utilization of ferromagnetic nanoparticles in hemodialysis therapy.

The in vitro utilization of biocompatible ferromagnetic nanoparticles (BFNs) in hemodialysis (HD), routinely used today for the treatment of end stage renal disease (ESRD), is introduced in this work. The proposed strategy is termed magnetically assisted hemodialysis (MAHD) and it aims to become a more efficient development of conventional HD. The method is based on the production of biocompatible ferromagnetic nanoparticles-targeted binding substances conjugates (BFNs-TBSs Cs) constructed of BFNs and specifically designed TBSs that should have high affinity and binding capacity for target toxic substances (TTSs) which must be removed from the ESRD patient subjected to HD. Antibodies or even specific proteins could serve as the TBS of the desired BFNs-TBSs Cs. The BFNs-TBSs Cs should be administered to the patient timely prior to the MAHD session so as to bind with the desired TTSs during their free circulation in the vascular network. Eventually, the complete BFNs-TBSs-TTSs structure can be selectively removed during the MAHD session by means of an external inhomogeneous magnetic field that is applied either at the dialyzer or at other collection point(s) along the blood circulation line of the dialysis machine. The advantages of MAHD over conventional HD regarding the patient's comfort and overall health status are discussed in detail among practical issues. To examine this proposition we employed Fe(3)O(4) and bovine serum albumin (BSA) as the BFN and the TBS constituents respectively, since they are both highly biocompatible. By means of x-ray diffraction, atomic force microscopy, circular dichroism spectropolarimetry, UV-vis spectrophotometry, SQUID magnetometry, and nuclear magnetic resonance we evaluated (i) the structural/morphological characteristics, (ii) the magnetic retraction efficiency, and most importantly (iii) the toxin binding affinity and capacity of both bare Fe(3)O(4) BFNs and Fe(3)O(4)-BSA Cs by performing in vitro experiments on specific TTSs. Homocysteine and p-cresol were chosen as representative TTSs and were investigated in great detail. The results obtained prove the in vitro applicability of the proposed MAHD method. Corrections were made to this article on 6 November 2007 (see figure 10 caption and lines 10 and 11 of page 11). The corrected electronic version is identical to the print version.

Samarium-153 and technetium-99m-labeled monoclonal antibodies in angiogenesis for tumor visualization and inhibition.

BACKGROUND: Angiogenesis is the development of new blood vessels from pre-existing ones. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic activators. Our studies are focused on the detection of VEGF by use of radiolabeled anti-endothelial monoclonal antibodies, which have the ability to localize in newly-formed vasculature of a cancerous origin. MATERIALS AND METHODS: The anti-endothelial monoclonal antibody VG76e was labeled with Samarium-153 and Technetium-99m. Biodistribution of the radiolabeled species was assessed in normal female Swiss mice, while tumor uptake was also evaluated. RESULTS: VG76e was labeled with 99mTc and 153Sm, resulting in a single product with a labeling yield of over 95%. Biodistribution studies showed non-specific uptake in any organ, with elimination via the hepatobiliary system. Finally, satisfactory tumor uptake was observed for both radiolabeled derivatives. CONCLUSION: Monoclonal antibodies raised against epithelial growth factors or their receptors, when labeled with appropriate radionuclides, may be a useful tool for early tumor detection and eventually for therapy.

Labeling of monoclonal antibodies with 153Sm for potential use in radioimmunotherapy.

The labeling of a monoclonal (anti-CEA) and a polyclonal (IgG) antibody with 153Sm has been investigated, using the bicyclic anhydride of DTPA (cDTPAa) as the chelating agent. The radiochemical study was performed using a combination of radioanalytical techniques (gel filtration, HPLC, ITLC-SG and SDS-PAGE). Optimization of factors affecting labeling (pH, Ab, Ab-DTPA concentration, etc.) leads to a labeling yield higher than 90%. Biodistribution studies in normal mice showed slow blood clearance and high uptake into the liver, kidney and lungs.

Novel oxorhenium and oxotechnetium complexes from an aminothiol[NS]/thiol[S] mixed-ligand system.

The simultaneous action of a bidentate aminothiol ligand, LnH, (n = 1: (CH3CH2)2NCH2CH2SH and n = 2: C5H10NCH2CH2SH) and a monodentate thiol ligand, LH (LH: p-methoxythiophenol) on a suitable MO (M = Re, 99gTc) precursor results in the formation of complexes of the general formula [MO(Ln)(L)3] (1, 2 for Re and 5. 6 for 99gTc). In solution these complexes gradually transform to [MO(Ln)(L)2] complexes (3, 4 for Re and 7, 8 for 99gTc). The transformation is much faster for oxotechnetium than for oxorhenium complexes. Complexes 1-4, 7, and 8 have been isolated and fully characterized by elemental analysis and spectroscopic methods. Detailed NMR assignments were made for complexes 3, 4, 7, and 8. X-ray studies have demonstrated that the coordination geometry around rhenium in complex 1 is square pyramidal (tau = 0.06), with four sulfur atoms (one from the L1H ligand and three from three molecules of p-methoxythiophenol) in the basal plane and the oxo group in the apical position. The L1H ligand acts as a monodentate ligand with the nitrogen atom being protonated and hydrogen bonded to the oxo group. The four thiols are deprotonated during complexation resulting in a complex with an overall charge of zero. The coordination geometry around rhenium in complex 4 is trigonally distorted square pyramidal (tau = 0.41), while in the oxotechnetium complex 7 it is square pyramidal (tau = 0.16). In both complexes LnH acts as a bidentate ligand. The NS donor atom set of the bidentate ligand and the two sulfur atoms of the two monodentate thiols define the basal plane, while the oxygen atom occupies the apical position. At the technetium tracer level (99mTc), both types of complexes, [99mTcO(Ln)(L)3] and [99mTcO(Ln)(L)2], are formed as indicated by HPLC. At high ligand concentrations the major complex is [99mTcO(Ln)(L)3], while at low concentrations the predominant complex is [99mTcO(Ln)(L)2]. The complexes [99mTcO(Ln)(L)3] transform to the stable complexes [99mTcO(Ln)(L)2]. This transformation is much faster in the absence of ligands. The complexes [99mTcO(Ln)(L)2] are stable, neutral, and also the predominant product of the reaction when low concentrations of ligands are used, a fact that is very important from the radiopharmaceutical point of view.