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Succinic dihydrazide (SDH), N-methyl-S-methyl dithiocarbazate (HDTCZ) and PEGylated N-methyl-S-methyl dithiocarbazate (HO2C-PEG600-DTCZ) are nitrido nitrogen atom donors employed for the preparation of nitride [M(N)]‐complexes (M = 99mTc and 188Re). This study aims to compare the capability and the efficiency of these three N3− group donors, in the preparation of [M(N)PNP]-based target-specific compounds (M = 99mTc, 188Re; PNP = aminodiphosphine). For this purpose, three different kit formulations (SDH kit; HO2C-PEG600-DTCZ kit; HDTCZ kit) were assembled and used in the preparation of [M(N)(cys~)(PNP3)]0/+ complexes (cys~ = cysteine derivate ligands). For each formulation, the radiochemical yield (RCY) of the [M(N)(~cys)(PNP3)] compounds, was determined by HPLC. The deviation of the percentage of RCY, due to changes in concentration of the N3− donors and of the exchanging ligand, was determined. For 99mTc, data clearly show that HDTCZ is the most efficient donor of N3−; however, SDH is the most suitable nitrido nitrogen atom donor for the preparation of [99mTc(N)(PNP)]-based target-specific agents with high specific activity. When HO2C-PEG600-DTCZ or HDTCZ are used in N3− donation, high amounts of the exchanging ligand (10−4 M) were required for the formation of the final complex in acceptable yield. The possibility to usemicrogram amounts of HDTCZ also in [188Re(N)] preparation (0.050 mg) reduces its ability to compete in ligand exchange reactions, minimizing the quantity of chelators required to obtain the final complex in high yield. This finding can be exploit for increasing the radiolabeling efficiency in [188Re(N)]-radiopharmaceutical preparations compared to the previously reported HDTCZ-based procedure, notwithstanding a purification process

Two series of novel tetrahedral copper(I) complexes comprising tridentate PNP-aminodiphosphines and tetradentate PN(X)P-substituted aminodiphosphines (X = O, S) have been prepared and characterized by conventional physico-chemical techniques. The first series includes ‘3 + 1’-type complexes comprising an aromatic PNP-aminodiphosphine and acetonitrile or triphenylphosphine. In the second series, the central amine function of the PNP-ligand was substituted with functionalized pendant arms containing ether, hydroxyl or thioether groups to enhance the chelation ability of the ligand. Fully coordinated neutral and cationic complexes were isolated. A preliminary study investigating both the labeling of 64Cu with the prototype PN(S)P ligand and the potential cytotoxic activity of the ‘cold’ [Cu(PN(S)P)][BF4] complex is reported.

We report on an efficient procedure for the preparation of [188Re(N)(PNP)]-based complexes (where PNP is diphosphinoamine) useful in the development of target-specific radiopharmaceuticals. The radiochemical yield of the compounds was optimized considering such reaction parameters as nature of the nitrido nitrogen donor, reaction times and pH level. The chemical identity of the 188Re agents was determined by high-performance liquid chromatography comparison with the corresponding well-characterized cold Re compounds. 188Re(N) mixed compounds have been evaluated with regard to stability toward transchelation with GSH and degradation by serum enzymes. The clearance of selected radiocompounds from normal tissues and their in vivo stability were evaluated in rats by biodistribution and imaging studies. [188Re(N)(cys∼)(PNP)]+/0 mixed-ligand compounds were efficiently prepared in aqueous solution from perrhenate using a multistep procedure based on the preliminary formation of the labile 188ReIII–EDTA species, which easily undergo oxidation/ligand exchange reaction to afford the [188ReV≡N]2+ core in the presence of dithiocarbazate. The final mixed-ligand compounds were obtained, at 100°C, by adding the two bidentate ligands to the buffered [188ReV≡N]2+ solution (pH 3.2–3.6). However, a relatively high amount of cys∼ ligand was required to obtain a quantitative radiochemical yield. The complexes were stable toward reoxidation to perrhenate and ligand exchange reactions. In vivo studies showed rapid distribution and elimination of the complexes from the body. No specific uptakes in sensitive tissues/organs were detected. A positive correlation of the distribution of the complexes estimated with biodistribution studies (%ID) and with micro-SPECT semiquantification imaging analysis (standard uptake values) was observed. These results s

This study presents the first application of a general procedure based on the use of the [Tc(N)Cl(PS) (PPh3)] species (PS is an alkyl phosphinothiolate ligand) for the preparation of Tc(N) target-specific compounds. [Tc(N)Cl(PS)(PPh3)] selectively reacts with an appropriate dithiocarbamate ligand (S^Y) to give [Tc(N)(PS)(S^Y)] compounds. 1-(2-Methoxyphenyl)piperazine, which displays a potent and specific affinity for 5HT1A receptors, was selected as a functional group and conjugated to the dithiocarbamate unit through different spacers (Ln). [99mTc(N)(PS)(Ln)] complexes were prepared in high yield (more than 90%). The chemical identity of 99mTc complexes was determined by high performance liquid chromatography comparison with the corresponding 99gTc complexes. All complexes were found to be inert toward transchelation with an excess of glutathione and cysteine. No notable biotransformation of the native compound into different species by the in vitro action of the serum and liver enzymes was shown. Nanomolar affinity for the 5HT1A receptor was obtained for [99mTc(N)(PSiso)L3] (IC50 = 1.5 nM); a reduction of the affinity was observed for the other complexes as a function of the shortening of the alkyl chain interposed between the dithiocarbamate and the pharmacophore. Negligible brain uptake was found from in vivo distribution data of [99mTc(N)(PSiso)L3]. The key finding of this study is that the complexes maintained good affinity and selectivity for 5HT1A receptors, and the IC50 value for [99gTc(N)(PSiso)L3] being comparable to the IC50 value found for WAY 100635. This result confirmed the possibility of preparing [99mTc(N)(PS)]-based targetspecific compounds without affecting the affinity and selectivity of the bioactive molecules for the correspond

An efficient procedure for the preparation of [188Re(N)(PNP)]-based complexes is reported. The radiochemical yield (RCY) of the compounds was optimized considering reaction parameters such as: nature of nitrido nitrogen donors, reaction times and pH. The chemical identity of 188Re-agents was determined by high-performance liquid chromatography comparison with the corresponding well characterized cold Re-compounds. The stability of the [188Re(N)(PNP)] complexes toward transchelation with glutathione and degradation by serum enzymes was assessed. The clearance of selected radiolabeled agents from normal tissues was evaluated in rats. [188Re(N)(cys∼)(PNP)]+/0 asymmetrical compounds were prepared in aqueous solution from perrhenate using a multistep procedure based on the preliminary formation of the labile 188ReIIIEDTA species which easily undergo to oxidation/ligand exchange reaction to afford in presence of DTCZ the [188ReV≡N]2+core. The final compounds were obtained by adding to the [188ReV≡N]int 2+ the two bidentate ligands. However, a relative high amount of cys∼ ligand was required to obtain a quantitative RCY. The complexes were stable toward reoxidation to perrhenate and ligand exchange reactions. In vivo studies showed a rapid distribution and elimination of the complexes from the body. No specific uptakes in radiation sensitive tissues/organs were detected. A positive correlation of the distribution of the complexes estimated with biodistribution studies (% injected dose) andwithmicro-single photon emission computed tomography semiquantification imaging analysis was observed. The low labeling efficiency limits but not excludes the possibility to apply [188Re(N)(PNP)]-technology to the preparation of target specific radiotherapeutical agents.

Aim: This study was focused on evaluating the applicability of a new labelling procedure to the preparation of Tc(N)‐based target specific compounds. The chemistry is based on the use of the [Tc(N)Cl(R2PS)(PPh3)] species (R2PS = alkyl‐phosphino‐thiolate ligand), which selectively reacts with an appropriate mono‐negative chelate, such as a dithiocarbamate (DTC), to give neutral [Tc(N)(R2PS)(DTC)] compounds. The 2‐methoxyphenylpiperazine (2‐MPP) pharmacophore, which displays a potent and specific affinity for 5HT1A receptors, was selected as functional group and conjugated to the dithiocarbamate unit through different spacers. Method: The synthesis of [99m/99gTc(N)(R2PS)(Ln)] complexes, and their in vitro stability as well as their biological in‐vitro and in‐vivo assays were investigated. Stability studies were performed by considering: i) stability toward transchelation with Cysteine and Glutatione ii) binding to the serum proteins; ii) stability in rat serum, human serum and rat liver homogenates. The in vitro affinity for the 5HT1A receptors of the technetium complexes was assessed by measuring the ability of the compounds to compete with [3H]‐8‐OH‐DPAT binding in isolated membranes from rat cerebral cortex. The biodistribution profile of the best radiolabeled compound and its in vivo stability were evaluated in Sprague‐Dawley rats. Results: [99mTc(N)(R2PS)(Ln)] complexes were prepared in high yield (>90%) using a multi‐step procedure. The chemical identity of 99mTc‐complexes was determined by HPLC comparison with the corresponding 99gTc‐complexes. All complexes were found to be inert toward transchelation with an excess of free Glutathione and Cysteine. No significant in vitro serum protein binding and no notable biotransformation of the native compound into different species by the in vitro action of the serum and liver enzymes were shown. Nanomolar affinities for the 5‐HT1A receptor were obtained for [99mTc(N)(PSiso)L3] (IC50 = 1.5 nM), a reduction of the affini

Rhenium(III) mixed-ligand complexes with tetradentate/monodentate (‘4 + 1’) coordination of the general formula [Re(NS3)(PRR’R’’)] (NS3 = tris-(2-mercaptoethyl)amine and derivatives thereof, PRR’R’’ = tertiary phosphine) are promising candidates for the development of rhenium-188 complexes for radiotherapeutic applications. In order to understand relationships between the structure of 188Re ‘4+1’ complexes and their in vitro stability we synthesized a series of rhenium model complexes using various combinations of NS3 derivatives and monodentate phosphorus(III) ligands and determined their stability at MBq activity level in human plasma. As an outcome of these investigations we are able to roughly estimate which ligands are suitable for a rational complex design with regard to high in vitro and in vivo stability. Subsequently, the most stable representative was investigated at GBq activity level showing no detectable degradation of the rhenium-188 complex in pharmaceutical preparation up to 2 hours after preparation. This stable 188Re ‘4+1’ complex was furthermore successfully conjugated to the model peptide arginine-tyrosine using the water-soluble N-hydroxy-sulfosuccinimidyl activated ester of the monodentate phosphine ligand. In summary, our results show the great potential of 188Re ‘4+1’ complexes for labelling biomolecules for therapeutic purposes.

In oder to understand relationships between the structure of rhenium-188 '4+1' complexes and their in vitro stability we synthesized a series of rhenium model complexes and determined their stability in human plasma. Rhenium-188 '4+1' complexes were synthesized using different monodentate phosphorus(III) ligands, including both lipophilic and hydrophilic tertiary phosphines and phosphites (188Re1-3) and unsubstituted tetradentate ligands and substituted NS3 ligands bearing a carboxyl group or an isopropyl amide were used as chelators (188Re4-6). As instability in aqueous solution leads always to perrhenate, the amount of 188ReO4- formed after 1h, 24 h and 48 h was determined by TLC. According to our findings 188Re4 was of sufficient stability and therefore used for further investigations. We tried to find out which physico-chemical parameter of the corresponding non-radioactive rhenium complexes Re1-6 may govern the formation of complexes of high in vitro stability. The water-soluble N-hydroxysulfosuccinimidyl activated ester 1 is a useful compound for the conjugation of exclusively water soluble biomolecules. Rhenium-188 labeling of the phosphine-arginine-tyrosine conjugate 2 as model compound was carried out using a labile 188Re(III)-EDTA intermediate. Based on this procedure peptides and proteins shall by labeld and tested in vivo.

Trigonal-bipyramidal Re(III) complexes with tetra-dentate/monodetate NS3/P ‘4+1’-coordination are inter-esting agents for the development of rhenium-188 com-plexes for potential therapeutic application. In order to understand relationships between the structure of rhenium-188 ‘4+1’ complexes and their in vitro stability we synthesized a series of rhenium model complexes 1, 2 and determined their stability in human plasma. The complexes were prepared as described in [1] using various combinations of NS3 derivatives and mono-dentate phosphorus(III) ligands. Since instability in aqueous solution always leads to the formation of perrhenate, we determined the amount of perrhenate formed after 1h, 24 h and 48 by TLC. By means of physico-chemical parameters of the corres-ponding non-radioactive rhenium complexes we tried to find factors which may govern the formation of complexes showing high in vitro stability. Finally, the most stable representative 3 was studied in 188Re labeling experiments with a phosphine-arginine-tyrosine model conjugate using the water-soluble N-hydroxysulfosuccinimidyl ester of the monodentate phosphine.

Development of new radiopharmaceuticals based on rhenium-188 depends on finding appropriate ligands able to give complexes with high in vivo stability. Rhenium(III) mixed-ligand complexes with tetradentate/monodentate (‘4 + 1’) coordination of the general formula [Re(NS3)(PRR’R’’)] (NS3 = tris-(2-mercaptoethyl)amine and derivatives thereof, PRR’R’’ = phosphorus(III) ligands) appear to be among the promising tools to achieve this goal. According to this approach, we synthesized and characterized series of rhenium model complexes. In vitro stabilities of the corresponding rhenium-188 complexes were determined by incubating 2-3 MBq or alternatively 37 MBq of the complexes in phosphate buffer, human plasma and rat plasma, respectively, at 22 °C or 37 °C, followed by checking the amount of 188ReO4- formed after 1 h , 24 h and 48 h by thin-layer chromatography. The rate of perrhenate formation varied over a wide range, depending primarily on the nature of the phosphorus(III) ligand. Physicochemical parameters of the corresponding nonradioactive rhenium complexes were analyzed in detail to find out the factors influencing their different stability and furthermore to design new substitution-inert ‘4 + 1’ complexes. Tolman’s cone angle of phosphorus(III) ligands and the lipophilic character of the inner coordination sphere were found to be crucial factors to build up stable rhenium ‘4 + 1’ complexes. Additional information useful to describe electronic and steric properties of these compounds were selected from electronic spectra (wavelength of the Re→S charge-transfer band), cyclovoltammetric measurements (E° of the ReIII/ReIV couple), and NMR investigations (31P chemical shift of coordinated P(III) ligands).

A new biomolecule labeling method that utilizes the [99mTc(N)(PNP)]2+m metal fragment is presented. Thus, a series of nitrido mixed-ligand M(V) complexes (M = 99mTc, 99gTc, Re), [M(N)(Ln)(PNP)], where Ln is the dianionic form of a dithiolate or substituted-dithiolate ligand and PNP is an aminodiphosphine, is described. 99mTc complexes can be prepared using either a two-step or a three-step procedure starting from generator-eluted pertechnetate through a pre-reduced mixture of [99mTc(N)] containing species, followed by contemporary or sequential addition of the relevant dithiolate and aminodiphosphine. The reaction of 2,3-dimercapto propionic acid (H2L1) with [Tc(N)(PNP)]2+ was investigated in detail. It was found that this bidentate ligand coordinated the metal fragment through the [S-,S-] donor atom pair, to yield neutral mixed-ligand complexes [99mTc(N)(L1)(PNP)] in high specific activity. The additional carboxylic functional group was not involved in metal coordination, thus remaining available for conjugation to target-specific molecules. Dithiolates incorporating pendant functional group(s) gave rise to a 1:1 diastereoisomeric mixture of syn-[M(N)(Ln)(PNP)] and anti-[M(N)(Ln)(PNP)] derivatives, depending on the relative orientation of the dithiolate substituent(s) with respect to the terminal nitrido group, and no isomeric conversion was detected. 99mTc species had been proven to be identical with the 99gTc complexes prepared at the macroscopic level by comparison of the corresponding radiometric and UV/vis HPLC profiles. Challenge experiments with cysteine or glutathione indicated that these physiological agents had no effect on the stability of this class of mixed-ligand 99mTc-complexes. Biodistribution studies in rats of selected 99mTc-complexes showed a rapid clearance from the blood and tissues

The nature of the heteroatom X incorporated in the five-member PXP-diphosphine spacer was found to play a primary unit role both in the stereochemical arrangement and overall stability of nitrido containing [M(N)(PXP)]2+ metal fragments (M = Tc, Re). Thus, by mixing PXP ligands with labile [Re(N)Cl4]- and [Tc(N)Cl2(PPh3)2] nitrido precursors in CH2Cl2/MeOH mixtures, a series of neutral [M(N)Cl2(PXP)] complexes (M = Tc, 1-5; M = Re, 8-9) was collected. In the resulting distorted octahedrons PXP adopted facial or meridional coordination, and combination with halide co-ligands produced three different stereochemical arrangements, i.e. fac,cis mer,cis and mer,trans, depending primarily on the nature of the diphosphine heteroatom X. When X = NH, mer,cis-[Tc(N)Cl2(PNP1)] 1 was the only isomer formed. Alternatively, when a tertiary amine nitrogen (X = NR; R = CH3, CH2CH2OCH3) was introduced in the spacer, fac,cis-[M(N)Cl2(PN(R)P)] complexes (M = Tc, 2, 3; M = Re, 8f) were obtained. Isomerization into the the mer,cis-[Re(N)Cl2(PN(R)P)], 8m, species was observed only in the case of rhenium when the tertiary amine group carried the less encumbering methyl substituent. Fac,cis-[Tc(N)Cl2(PSP)], 4f, was isolated in the solid state when X = S, but mixture of fac,cis-[Tc(N)Cl2(PSP)] and mer,trans-[Tc(N)Cl2(PSP)], 4m, isomers was found in equilibrium in the solution state. A similar equilibrium between fac,cis-[M(N)Cl2(POP)] (M = Tc, 5f; M = Re, 9f) and mer,trans-[M(N)Cl2(POP)] (M = Tc, 5m; M = Re, 9m) species was detected in POP containing complexes. The molecular structure of all of these complexes were assessed by means of conventional physico-chemical techniques including multinuclear NMR spectroscopy and X-ray diffraction analysis of representative m

A novel type of mixed-ligand Tc(III) complexes, [Tc(SCH2CH2-E-CH2CH2S)(PR2S)] (E = S, N(CH3); PR2S = phosphinothiolate with R = aryl, alkyl) is described. These "3+2"-coordinated complexes can be prepared in a two-step reduction/substitution procedure via the appropriate chloro-containing oxotechnetium(V) complex [TcO(SES)Cl] (E = S, N(CH3). Tc(III) compounds have been fully characterized both in solid and solution states and found to adopt the trigonal-bipyramidal coordination geometry. The equatorial trigonal plane is formed by three thiolate sulfurs atoms, whereas the phosphorus of the bidendate P,S ligand and the neutral donor of the tridendate chelator occupy the apical positions. The 99Tc(III) complexes have been proven to be identical with the 99mTc-agents prepared at the no-carrier-added level by comparison of the corresponding UV-Vis and radiometric HPLC profiles. Challenge experiments with glutathione clearly indicated that this tripeptide has no effect on the stability of the 99mTc complexes in solutions. Biodistribution studies were carried out in rats at 5 and 120 minutes post injection. The substituents at the bidendate P,S ligand significantly influenced the biodistribution pattern. Remarkable differences were observed especially in brain, blood, lungs and liver. All the complexes were able to penetrate the blood-brain-barrier of rat brain and showed a relatively fast washout from the brain.

Neutral nitrido M(V) heterocomplexes of the type [M[N)(PNP)(S-S)] (M=99Tc, Re, 99mTc; PNP=diphosphinoamine; S-S=dithiolate) have been prepared and fully characterised at macroscopic as well as nca levels. Chirality introduced at the dithiolate framework by insertion of a carboxylic group induces the formation of syn- and anti- diastereoisomers.

Der Zugang zu diagnostisch relevanten Komplexverbindungen des Tc-99m wird in der Kopplung des Metalls an biologisch aktive Moleküle gesucht. Bisher dominieren Tc(V)-Chelate mit charakteristischem [Tc=O]3+ core. Die hohe Polarität der Metall-Sauerstoff-Bindung sowie die Austauschfähigkeit der Liganden in vivo stimulieren die Suche nach stabilen Systemen auf niederen Oxidationsstufen. Wir beschreiben hier zwei neue Typen von Tc(III)-Komplexen. Sie erfüllen die Forderung nach niedriger Polarität und hoher in-vivo-Stabilität. Gekoppelte Substitutions-Reduktionsreaktionen von Tc(V)-Verbindungen 1 mit zwei-zähnigen PS-Liganden führen zu Tc(III)-Komplexen der Struktur 2. Die Reduktion von Pertechnetat mit tertiären Phosphinen in Gegenwart des tripodalen Liganden Tris(2-mercaptoethyl)amin ergibt Gemischtligandkomplexe 3 mit NS3/P-Koordination. Der monodendate Phosphinligand kann in einfacher Weise durch Isocyanide unter Bildung der Spezies 4 substituiert werden. Die neuen Tc(III)-Chelate 2 und 4 zeichnen sich durch eine hohe Variabilität hinsichtlich des zu koppelnden Biomoleküls (R) aus. Challenge-Experimente mit Glutathion unter physiologischen Bedingungen belegen die hohe in-vivo-Stabilität der Verbindungen.

The most frequently used technetium compounds suitable for coupling the metal to biologically active molecules are square-pyramidal oxo complexes based on tetradentate N2S2 ligands (1). Properties and thus the in vivo behaviour of such complexes are strongly influenced by the presence of the quite polar [Tc=O]3+ unit. Whether such a polarity is beneficial or not is not obvious and depends on the requirements for specific radiotracers, e.g. for receptor-targeting agents or for metabolic tracers. Another crucial point in tracer design is the stability of the respective Tc chelate towards ligand exchange in vivo. Recently it was shown that the applicability of the "3+1" mixed-ligand Tc(V) chelates is limited because of their reaction with glutathione in vivo (2,3). So there is a considerable interest in alter-native chelate systems that combine lower polarity with enhanced in vivo stability. Such systems are based on oxo-free Tc cores at lower oxidation state. Here we describe novel types of Tc(III) mixed-ligand complexes. Both chelate systems fulfil the requirements for a lipophilic, non-polar building block stable against ligand exchange reaction in vivo. They offer the advantage of high versatility in conjugating biomolecules as described for the "3+1"-series (4).

Five-coordinate oxotechnetium(V) mixed-ligand complexes [TcO(SES)(S-p-C6H4-OMe)], where SES is a tridentate dithiolato fragment of the type -S(CH2)2E(CH2)2S- (E = O, 1; E = S, 2; E = NMe, 3) are converted via reduction-substitution reactions in the presence of PMe2Ph into the corresponding five-coordinate Tc(III) complexes [Tc(SES)(S-p-C6H4-OMe)(PMe2Ph)] (E = O, 4; E = S, 5; E = NMe, 6). Rearrangement of the original square pyramidal '3 + 1' oxo-species to the trigonal bipyramidal '3+1+1' Tc(III) complexes occurs by placing the three thiolate donors on the basal plane, the phosphine phosphorus and the heteroatom of the tridentate ligand at the apexes of the bipyramid. These Tc(III) complexes are diamagnetic species, thereby allowing multinuclear NMR characterization in solution, which confirm their structures to be identical to those observed in the solid state via X-ray determinations.

To date most Tc compounds suitable for coupling to biologically active molecules, e.g. CNS receptor targeted agents, are square-pyramidal complexes of the oxo ion [Tc=O]3+. However, the properties and thus the in vivo behaviour of such complexes are strongly influenced by the quite polar Tc=O unit offering a free position trans to the oxo ligand for further reaction in vivo. The stricter the requirements for specific agents, the more important is the question whether such a polarity is beneficial or not. Alternatives are oxo-free lower oxidation states. Here we report on prototypic trigonal-bipyramidal mixed-ligand complexes 1, 2 and 3 which are neutral, non-polar and robust since they contain sterically well shielded oxo-free Tc(III). We describe the syntheses and structural analyses of 99Tc complexes with simple co-ligands serving as models for functionalized derivatives, the no-carrier-added preparation of the analogous 99mTc-complexes as well as their stability in aqueous solution and in plasma, and their biodistribution in rats.