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Cyclic and acyclic ligands containing the hydroxypyridinone (HOPO) moiety as donor group are known as strong coordinating compounds for a wide variety of metal ions. Based on the diaza-crown[18]ether Kryptofix K22, five different tendentate ligands were prepared using 1,2-HOPO, 1,2,3-HOPO and 2,3-Me-HOPO as additional binding moieties. The diaza-crown ether basic skeleton was furnished with two primary amine functions and subsequently reacted with the respective HOPO acids or the HOPO acid chlorides to obtain the desired HOPO derivatives in two synthesis steps after final deprotection. All compounds were evidenced by NMR and MS analyses.

Macropa-PSMA-Konjugate, die zur zielgerichteten Alphatherapie mit Ac-225 dienen, werden mit einem Albuminbinder verknüpft. Neben der Verbesserung des pharmakologischen Verhaltens in vivo bietet die albuminbindende Einheit die Grundlage eines theranostischen Ansatzes. Neben der Komplexierung des Alphaemitters Ac-225 wird zusätzlich die Einführung des SPECT-Nuklids I-123 im gleichen Molekül ermöglicht. Vorteilhaft sind die milden Bedingungen der Radioiodierung sowie eine Halbwertszeit (t1/2 = 13,2 h), welche die Bildgebung länger zirkulierender Substanzen ermöglicht.

Complexes of macrocyclic ligands are commonly utilized in many medicinal applications, such as MRI contrast agents and radiopharmaceuticals. Due to their thermodynamic stability and kinetic inertness, they overcome toxicity of free heavy metal ions and, thus, enable their use in vivo. However, there is no “general” ligand suitable for any metal ion in the periodic table and, thus, useful chelator should be specially designed for each metal ion. Among metal radionuclides, there is an increasing interest in those of large elements (i.e. large metal ions) to be used for both, radiotherapy and imaging. The 18-memberred macrocycles are suitable scaffolds for the utilizations. This contribution deals with chemistry and radiochemistry of H4pyta (18-py2N4Ac4), a parent rigidified macrocycle suitable for large metal ions. Although the ligand was synthesized in mid-nineties, there are almost no data focused on its possible utilization in radiopharmaceutical context.[2] The ligand forms thermodynamically stable complexes with Ln(III) ions. Its ten donor atoms completely wrap the large Ln(III) ions but one pendant arm is not coordinated for small Ln(III) ions. Thus, the early Ln(III) ions form more stable complexes. The Ln(III)-H4pyta complexes are 2–3 orders of magnitude more kinetically inert than those of H4dota and, thus, they will be fully stable in vivo. Radiolabeling of H4pyta with 133La and 177Lu is, at different pH’s and temperatures, comparable with that of H4dota. Challenging experiments showed that these radiolabeled H4pyta complexes are fully resistant to transmetallation and transchelation, and show very high stability in human blood serum stability. For larger 133La, the results are much better if compared with data obtained for H4dota and macropa in the parallel experiments. For smaller 177Lu, the results are also more promising compared to those of H4dota and its analogues. This ligand shows that the H4pyta (18-py2N4Ac4) scaffold offers new possibilities for de

Ziel/Aim: Der Einsatz von Actinium-225 mit dem DOTA-Konjugat PSMA-617 befindet sich in der klinischen Entwicklung. DOTA ist jedoch nicht ideal, da hohe Temperaturen für die Markierung benötigt werden und der Komplex in vivo nicht stabil genug ist. Ziel ist es, einen besseren, macropa-basierten Chelator zu finden und mit von PSMA-617 abgeleiteten Bindungsvektoren zu verknüpfen. Methodik/Methods: Es wurden 4 verschiedene Konjugate hergestellt, welche sich hinsichtlich der Anzahl der Bindungsmotive sowie ihrer Fähigkeit, an Albumin zu binden, unterscheiden. Die Stabilität der markierten Komplexe wurde über 10 Tage in Puffer und Serum untersucht. Weiterhin wurden Bindungsaffinitäten und Koloniebildungsassays an PSMA-positiven LNCaP-Zellen durchgeführt. Alle Konjugate wurden hinsichtlich ihrer Organverteilung in LNCaP-Tumor-tragenden SCID-Mäusen untersucht. Ergebnisse/Results: Alle 4 Konjugate wurden unter milden Bedingungen radiomarkiert und zeigen bei Zugabe von Gentisinsäure die höchste Stabilität über 10 Tagen. Die Bindungsaffinitäten der Konjugate wurden als KD-Werte bestimmt und liegen zwischen (10-40) nM. Die Untersuchungen zum Zellüberleben zeigen einen therapeutischen Vorteil der bivalenten gegenüber den monovalenten Konjugaten. Alle vier Konjugate erreichten in unterschiedlichen Zeitspannen (bis 5 d p.i.) Tumoranreicherungen von (12-85) %ID/g. Unerwünschte Anreicherungen traten v. a. bei dem dimeren, albuminbindenden Konjugat in Form von Akkumulationen (Leber, Milz, Nieren) auf. Schlussfolgerungen/Conclusions: Die Verwendung von macropa-basierten Konjugaten für die Therapie mit Actinium‑225 oder die zukünftige Bildgebung mit Lanthan‑133 ist vorteilhaft, da die gebildeten Komplexe stabiler sind und damit eine Anreicherung der Aktivität in off-Target-Regionen verringert werden kann. Weiterhin ergibt sich ein Vorteil im Handling, da die Reaktionen schnell und bei Raumtemperatur durchgeführt werden können, was die spätere Umsetzung in eine GMP-gerechte Herstellung

Objectives Theranostic concepts and the usage of alpha-particle emitting radionuclides belong to the emerging fields of radiopharmaceutical sciences. Especially and due to its excellent complexation properties, the chelator macropa (mcp)[1,2] was reported to be a suitable complexing agent for 225Ac conjugates[3] and as macrocycle a highly promising starting point for the development of 223/224Ra chelators as well. To follow the theranostic approaches with these alpha emitters, due to their chemical similarities 131Ba[4] for SPECT and 133La[5] for PET are available as diagnostic radionuclide surrogates of radium and actinium, respectively. In our recent study, we aim to establish a new workflow to evaluate and predict the complex stability of new chelating systems by obtaining both protonation constants for respective ligands and association constants for their metal complexes starting with mcp. Methods As a prerequisite for the calculation of stability constants (log(K)), the protonation constants (pKa) of mcp were determined by pH‑dependent 1H NMR studies in the first step. Based on these obtained data, the europium-mcp-complex was examined using time-resolved, laser-induced luminescence spectroscopy (TRLFS) and the log(K) was calculated as reference stability constant. Additionally, Eu-TRLFS was used to determine the Eu-mcp species during titration and pH-dependency of the complexation. The method of isothermal titration calorimetry (ITC) was used to measure the complex stability constants of La-, Ba-mcp-complex, as well as Eu-mcp-complex to value the results in comparison with TRLFS data. The evaluation and calculation of the log(K) values were carried out by parallel factor analysis (PARAFAC). Results Depending on the protonation ability of the functional groups found in mcp (amines and carboxylates), pKa values were obtained as well as the log(K) values for the respective mcp-complexes with Ba, Eu and La and are shown in Table 1. Conclusion By combining 1H-NMR

Das prostataspezifische Membranantigen (PSMA) ist ein hoch attraktives biologisches Target für die molekulare Bildgebung und für die zielgerichtete Radionuklidtherapie bei kastrationsresistentem Prostatakrebs (mCRPC), da es in allen Phasen der Erkrankung bei den Tumorzellen expremiert ist. Somit bieten hoch affine, PSMA-bindende Biomoleküle eine exzellente Basis zur Entwicklung von Radiokonjugaten für die Therapie mit Alphastrahlern. Aufgrund seiner hervorragenden kernphysikalischen Eigenschaften wird zunehmend Actinium-225 für diese Zwecke in der Nuklearmedizin verwendet. Das dreiwertige Kation Ac3+ muss dazu in einem Chelator hoch stabil komplexiert werden. Um das zu realisieren, wird der Chelator macropa verwendet, welcher eine sehr hohe In-vivo-Stabilität zeigt und Radiomarkierungen unter sehr milden Bedingungen zulässt. Dieser wird mit einer bzw. zwei PSMA-bindenden Einheiten modifiziert, um den 225Ac-Komplex zu den Tumorzellen zu bringen. Zunächst wurden zwei Actinium-225-Radiokonjugate entwickelt und deren pharmakologische Eigenschaften, z.B. die Bindung an Tumorzellen, Anreicherung im Tumor (ca. 10%ID/g für beide Konjugate) und in anderen Organen überprüft. Hier wurde ein ähnliches Verhalten festgestellt, vergleichbar mit dem Standardkonjugat 225Ac-PSMA-617. Die beiden 225Ac-Radiokonjugate wurden weiterentwickelt und mit einem Albumin-Binder (4-Iodphenylbutyrat) versehen. Dadurch wurde eine längere Blutverweilzeit, kombiniert mit einer höheren Aufnahme in den Tumor, erreicht (bis 50%ID/g Monomer, bis 120 %ID/g Dimer), um die Effektivität der Radiotherapie zu erhöhen.

Introduction: Radiolabeled prostate-specific membrane antigen (PSMA) ligands have been successfully developed in recent years for diagnosis and therapy of prostate cancer [1]. A potential new PSMA radioligand would, after in vitro characterization, normally be further evaluated in small animals. However, the use of micro-physiological systems (MPS), an organ-on-chip technology in which human cell spheroids and organoids are treated in a circuit under defined conditions [2,3], should lead to a reduction of animal experiments. In the present study, experiments with the novel 64Cu-labeled PSMA antagonist FR52 were performed using 2D or 3D cultures of LNCaP cells in MPS chips. Our aim was to determine binding parameters (Kd, Bmax) in the MPS chip and to compare them with the results of assays in well plates (WP). Methods: Derived from the compound PSMA-617 [4], instead of DOTA a cyclohexanediamine-triazole (CHDT) chelator was conjugated, resulting in the conjugate FR52 (in-house synthesis). FR52 was radiolabeled with 64Cu (molar activity: 14.2 ± 2.2 MBq/nmol). For the assays, 5 x 104 LNCaP cells were placed in 3 wells of an MPS module (à 0.2 cm2), either as monolayer or as spheroids and cultured for 2 to 7 days. The MPS chips include an integrated micropump, driven by a controller, that generates a pulsatile flow of media that runs through the capillaries and keeps the cells alive [5] (Fig. 1). After blocking of adjacent cell samples (PSMA pharmacophore 0.8 mM; 30 min) [64Cu]Cu-FR52 (6 to 200 nM) was added to all samples and pumped through the MPS for 10 min (volume: 0.1 mL/well; 80 bpm; volume flow 6.4 µL/s). After PBS washing and exposing the MPS chips to an imaging plate, the cells were lysed and in vials measured in a gamma counter. Evaluation was processed with the analysis program GraphPad Prism. Results: Saturation assays with [64Cu]Cu-FR52 in MPS revealed similar high binding affinities (Kd) for LNCaP monolayer and spheroids (MPS-monolayer Kd: 33 ± 6 nM; MPS-sph

Heavy alkaline earth metals offer radionuclides which are promising candidates for radiopharmaceutical applications like barium-131 for diagnosis or radium-223/-224 – with similar properties to barium – for targeted alpha-particle therapy. However, there is a lack of suitable chelation agents especially for these metal ions. A series of calix[4]crown-6 derivatives with perfluoroalkylsulfonylcarboxamide functions (RF = CF3, C2F5, i-C3F7, n-C4F9) was synthesized to serve as cage-like chelators for a strong complexation of Ba2+ and Ra2+. These functional ligands are deprotonated even at slightly acidic pH due to the intense electron withdrawing effect of the sulfonamide group. The obtained ligands were easily converted the desired barium complexes as well as into calix-crown compounds containing two sodium ions. DFT calculation methods were used to discover either the binding behavior of the metal ions with the desired ligands as well as the influence of the different donor groups from the chelating moiety of the calixarenes with respect to different pH. Radiolabeling procedures with the radionuclides barium-133 and radium-224 as [133Ba]BaCl2 and [224Ra]Ra(NO3)2 were performed to determine association constant values between 4.1 and 8.2 for the appropriate M2+ complexes using a two-phase extraction procedure. A stability test using physiological Ca2+ solution showed a minor release of approx. 1-7% of the central ions (Ba2+ respectively Ra2+) from the complexes.

To follow the design of in vivo stable chelating systems for radiometals, a concise and straightforward method toolbox was developed combining NMR, ITC, and Eu-TRLFS. For this purpose, the macropa chelator was chosen and Lu3+, La3+, Pb2+, Ra2+, and Ba2+ as radiopharmaceutically relevant metal ions. They differ in charge (+2, +3) and coordination properties (main group vs. lanthanides). 1H NMR was used to determine four pKa values (carboxylate functions: 2.40 and 3.13; amine functions: 6.80 and 7.73). Eu-TRLFS was used to validate the exclusive existence of the Mn+:ligand 1:1 complex in the chosen pH range. ITC measurements were accomplished to determine the resulting stability constants of the desired complexes with log K values ranging from 18.5 for the Pb-mcp complex to 7.3 for the Lu-mcp complex. DFT-calculated structures nicely mirror the complexes’ order of stabilities by bonding features. Radiolabeling with macropa using ligand concentrations from 10–3 M to 10–6 M pointed out the complex stability (133La > 131Ba > 177Lu).

Ziel: macropa-PSMA-Konjugate mit Albuminbinder zur zielgerichteten Alphatherapie mit Actinium-225 wurde innerhalb der Arbeitsgruppe bereits erfolgreich synthetisiert und in vitro und in vivo untersucht. Die dabei eingeführte albuminbindende Einheit bietet die Grundlage zur Erarbeitung eines theranostischen Ansatzes indem, neben der Komplexierung des Alphaemitters Actinium-225, die Bindung von Iod-123 als leicht zugängliches SPECT-Nuklid im gleichen Molekül ermöglicht wird. Vorteilhaft sind die milden Markierungsbedingungen sowie eine passende Halbwertszeit des Iodisotopes (t1/2 =13,2 h), welche die Bildgebung länger zirkulierender Substanzen ermöglicht. Methoden: Die Synthese der peptidomimetischen Ausgangsverbindungen erfolgte angelehnt an die Vorarbeiten durch mehrstufige Peptidkupplungen und die Anbringung des Chelators mittels Cu-katalysierter Click-Reaktion. Anstelle der 4-(p-Iodphenyl)buttersäure wurde der entsprechende Zinnprecursor eingeführt, um die Verbindung durch eine elektrophile, aromatische Substitution mit Iod-123 markieren zu können. Ergebnisse: Durch die erfolgreiche Entwicklung einer Methode zur Iodierung der synthetisierten Zinnprecursoren mit hohen radiochemischen Ausbeuten konnten schließlich die einfach bzw. doppelt PSMA-gebundenen Verbindungen [123I]I-mcp-M-alb-PSMA und [123I]I-mcp-D-alb-PSMA hergestellt werden. Nach der Iodierung der Verbindungen wurde in vitro der Einfluss des 139La-Komplexes als auch der des unkomplexierten Chelators an der PSMA-positiven Zelllinie LNCaP untersucht. Schlussfolgerungen: Die Einführung von Iod-123 in die bereits untersuchten Derivate mit stabilem Iod-127 ergab keine Änderungen der biologischen Ergebnisse. Damit ergibt sich der neue Aspekt des theranostischen Nutzens als hybrides Radiopharmakon mit einer weiteren Markierungsstelle in der Peripherie des eigentlichen Liganden.

The prostate-specific membrane antigen (PSMA) is expressed in approx. 80% of prostate cancer at considerably higher levels in tumor cells compared to healthy tissues. Its upregulation occurs in all stages of the disease. Therefore, PSMA has emerged as an attractive target for molecular imaging and especially targeted radionuclide therapy (endoradiotherapy) of metastatic castration-resistant prostate cancer (mCRPC). Due to its favorable nuclear physical properties, actinium-225 belongs to the clinically applied alpha emitters for therapeutic applications. More than ten clinical trials are displayed for actinium-225-DOTA-based conjugates. A trial using [225Ac]Ac-PSMA-617 as the IMP in men with PSMA-positive prostate cancer is projected to start. Actinium-225 can be chelated with the standard DOTA chelator, but requires rather harsh labeling conditions such as high temperatures (80–95°C) or microwave assistance. Alternatively, with the complexing agent macropa mild labeling conditions, low reaction temperature and high in vivo stability can be realized owing to its perfect characteristics for Ac3+ chelation. Moreover, to further improve the tumor uptake for future targeted alpha therapy, albumin binding units based on 4-iodophenylbutyrate were introduced to increase the tumor uptake. Two 225Ac-radioconjugates with albumin binders were prepared, either containing one or two PSMA-binding units (based on PSMA-617), the second one to raise the binding affinity to the PSMA receptor. The synthetic approach consisted of the functionalization of one or two picolinic side arms with a clickable reactive site for the connection of the vector molecules. The azide-functionalized PSMA derivative, which also contains the albumin binder, was prepared using standard peptide coupling conditions. Cell culture-based studies regarding radioligand affinity and clonogenicity were performed using PSMA-positive LNCaP cells and the biodistribution of the radioconjugates wa

Objectives The prostate-specific membrane antigen (PSMA) is expressed in most cases of prostate cancer at considerably higher levels in tumor cells compared to healthy tissues [1-3] and its upregulation occurs in all stage of the disease [4, 5]. Therefore, PSMA has emerged as an attractive target for molecular imaging and especially targeted radionuclide therapy (endoradiotherapy) of metastatic castration-resistant prostate cancer (mCRPC), given the example of [177Lu]Lu-PSMA-617. We recently described the synthesis and in-depth characterization of PSMA radioligands for targeted alpha therapy with actinium-225 [6]. Our present study aimed at the design, synthesis and preclinical evaluation of albumin-binding PSMA ligands in order to optimize their tissue distribution profile and to improve their pharmacokinetic properties. Methods Two novel compounds were prepared by combining a macropa chelator with one or two lysine-ureido-glutamate–based PSMA-binding entities equipped with 4-(p-iodophenyl)butyrate residues. The albumin-binding properties of the 225Ac-labeled conjugates were investigated in vitro using mouse, rat and human serum. The specific interaction of both compounds with human serum albumin was confirmed by microscale thermophoresis. Cell culture-based studies regarding radioligand affinity and clonogenicity were performed on PSMA-positive LNCaP cells. The biodistribution of the radioconjugates was analyzed in LNCaP tumor-bearing mice with ensuing investigation of tissue toxicity by histological examinations. Results Radiolabeling of both PSMA ligands with 225Ac was achieved at up to 5 MBq/nmol with >99% radiochemical purity. In vitro assays confirmed considerable binding of the radioligands to mouse, rat and human serum proteins. Cell binding and survival studies revealed a higher cell binding affinity and an improved cell killing efficiency for the radioconjugate with two PSMA-binding entities compared to the derivative with only one targeting motif. Biodis

The increasing application of positron emission tomography (PET) in nuclear medicine has stimulated the extensive development of a multitude of novel and versatile techniques to introduce fluorine-18 especially for the radiolabelling of biologically or pharmacology active molecules. Taking into consideration that the introduction of fluorine-18 (t1/2 = 109.8 min) mostly proceeds under harsh conditions, radiolabelling of such molecules represents a challenge and is of enormous interest. Ideally, it should proceed in a regioselective manner under mild physiological conditions, in an acceptable time span, with high yields and high specific activities. Special attention has been drawn to 2-fluoroethyl and 3-fluoropropyl groups, respectively, which are often the active site of radiofluorinated compounds. Precursors containing an ammonium leaving group which consists of a strained azetidinium or aziridinium moiety can help to overcome these obstacles leading to a mild introduction of [18F]fluoride with high radiochemical yields.

The combination of conventional chemotherapeutic drugs with radionuclides or external radiation is discussed for a long period of time. The major advantage of a successful combination therapy is the reduction of severe side effects by decreasing the needed dose and simultaneously increasing therapeutic efficiency. In this study, pUC19 plasmid DNA was incubated with the cytostatic drug cisplatin and additionally irradiated with 99mTc, 188Re and 223Ra. DNA damages, such as single- and double strand breaks were determined by agarose gel electrophoresis. The threshold concentration value of cisplatin, which was tolerated by pUC19 plasmid DNA was determined to be 18-24 nM. Nevertheless, even at higher dose values (>100 Gy) and simultaneous incubation of cisplatin to 200 ng plasmid DNA, no significant increase in the number of induced single- and double-strand breaks was obtained, compared to the damage solely caused by the radionuclides. We thereby conclude that there is no direct dependence of the mechanism of strand break induction to the absence or presence of platinum atoms attached to the DNA. Reported increasing DNA damages in therapy approaches on a cellular level strongly depend on the study design and are mainly influenced by repair mechanisms in living cells. Nevertheless, the use of radioactive cisplatin, containing the Auger electron emitter 191Pt, 193mPt or 195mPt, is a bright prospect for future therapy by killing tumor cells combining two operating principles: a cytostatic drug and a radiopharmaceutical at the same time.

Targeted Alpha Therapy is a research field of highest interest in specialized radionuclide therapy. Over the last decades, several alpha-emitting radionuclides have entered and left research topics towards their clinical translation. Especially, 225Ac provides all necessary physical and chemical properties for a successful clinical application, which has already been shown by [225Ac]Ac PSMA 617. While PSMA 617 carries the DOTA as the complexing agent, the chelator macropa as macrocyclic alternative provides even more beneficial properties regarding labeling and complex stability in vivo. Lanthanum-133 is an excellent positron-emitting diagnostic lanthanide to radiolabel macropa-functionalized therapeutics, since 133La forms a perfect matched theranostic pair of radionuclides with the therapeutic radionuclide actinium-225 which itself can optimally be complexed by macropa as well. 133La was thus produced by cyclotron-based proton irradiation of an enriched 134Ba target. The target (30 mg of [134Ba]BaCO3) was irradiated for 60 min at 22 MeV and 10-15 µA beam current. Irradiation side products in the raw target solution were identified and quantified: 135La (0.4%), 135mBa (0.03%), 133mBa (0.01%) and 133Ba (0.0004%). The subsequent work-up and anion-exchange-based product purification process took approx. 30 min and led to a total amount of (1.2 – 1.8) GBq (decay-corrected to EOB) of 133La, formulated as [133La]LaCl3. After complete decay of 133La, a remainder of ca. 4 kBq of long-lived 133Ba per 100 MBq of 133La was detected and rated as uncritical regarding personal dose and waste management. Subsequent radiolabeling was successfully performed with previously published macropa-derived PSMA inhibitors at a micromolar range (quantitative labeling at 1 µM) and evaluated by radio-TLC and radio-HPLC analyses. The scale up to radioactivity amounts that are needed for clinical application purposes would be easy to achieve by increasing target mass, beam current and/or irradi

We recognize 131Ba as a SPECT radionuclide, which provides a diagnostic match for the therapeutic alpha-emitting radionuclides 223Ra and 224Ra. The gamma-emitter barium-131 (t½ = 11.5 d) decays with an energy of 123.8 keV (30% intensity) of the first decay via cesium-131 (t½ = 9.7 d) to stable xenon-131, each by electron capture. Our aim was to develop of a straightforward resin-based radiochemical separation to yield 131Ba with high radionuclide purity. Furthermore, the radiolabeling of the complexing agent macropa with [131Ba]Ba2+ using mild labeling conditions was intended. For this purpose, different TLC systems as reaction control were utilized. The radiopharma-cological characterization of 131Ba-labeled macropa in comparison to uncomplexed [131Ba]Ba2+ was carried out in healthy mice, including biodistribution and small animal SPECT/CT experiments.

With the exception of beryllium, every alkaline earth metal is characterized by a calcimimetic behavior. Thus, in vivo biodistribution mostly occurs in form of a massive accumulation in bone tissues, consisting of hydroxyapatite to a major extent. Apart from the lightest elements beryllium and magnesium, animal studies and human studies regarding the overall in vivo behavior were carried out by using radioisotopes of the elements calcium, strontium, barium, and radium. To date, only strontium with its radioisotopes gained importance for applications in nuclear medicine, mainly for pain-reducing and palliative treatment of bone metastases. In contrast, barium isotopes can be useful imaging agents and possible diagnostic analogues for theranostic approaches. This review focuses on the characteristic and chemical behavior of barium compounds, possible radioactive barium isotopes for future applications in nuclear medicine and radiopharmacy as well as recent results regarding barium-131 as diagnostic match for radium isotopes used in targeted alpha therapy.

Currently, targeted alpha therapy is one of the most investigated topics in radiopharmaceutical cancer management. Especially, the alpha emitter 225 Ac provides excellent nuclear properties and is gaining increasing popularity for the treatment of various tumor entities. We herein report on the synthesis of two universal 225Ac-chelators for mild condition radiolabeling and binding sites to conjugate biomolecules via the copper-mediated click chemistry. A convenient radiolabeling procedure was investigated as well as the complex stability proved for both chelators and two PSMA-targeting model radioconjugates. Studies regarding affinity and cell survival were per-formed on LNCaP cells followed by biodistribution studies, which were performed using LNCaP tumor-bearing mice. High efficiency radiolabeling for all conjugates was demonstrated. Cell binding studies revealed a fourfold lower cell affinity for the PSMA conjugate with one targeting vector compared to the conjugate owing two targeting vectors. Additionally, these differences were verified by in vitro cell survival evaluation and biodistribution studies, both showing a higher therapeutic efficiency for the same dose on a cellular leve, a higher tumor up-take (15%ID/g) and a rapid whole body clearance after 24 hours. The synthesized chelators will overcome obstacles of lacking stability and worse labeling needs regarding 225Ac complexation using the DOTA chelator. Moreover, the universal functionalization expands the coverage of these chelators in combination with any sensitive bio(macro)molecule, thus improving treat-ment of any addressable tumor target.

Von Erdalkalimetallionen außer Beryllium ist bekannt, dass sie ein calcimimetisches Verhalten zeigen. Damit ist ihr Schicksal in vivo vorgezeichnet, das in einem beträchtlichen Maße durch den Einbau in Knochengewebe, welches zum Hauptteil aus Hydroxylapatit besteht, charakterisiert ist. In diesem Sinne wurde auch die Verwendung von Radionukliden dieser Elemente als Knochensucher forciert. Mit Ausnahme von Beryllium und Magnesium wurden Tierexperimente und Humananwendungen mit Radionukliden von Calcium, Strontium, Barium und Radium durchgeführt, wobei bis heute lediglich Strontium und Radium, in der Hauptsache als Therapienuklide zur palliativen Behandlung von Knochenmetastasen, Eingang in nuklearmedizinische Routineanwendungen gefunden haben. In diesem Übersichtsartikel werden die Radionuklide des Bariums vorgestellt, sowie deren Herstellung und Verwendung. Aktuelle Forschungsergebnisse mit Radionukliden des Bariums in Radiopharmazie und Nuklearmedizin werden präsentiert.

We understand 131Ba as a radionuclide, which enables imaging by SPECT in nuclear medicine and provides a diagnostic match for the therapeutic alpha-emitting radionuclides 223Ra and 224Ra. Recently, we reported on a sufficient production route for 131Ba by irradiating a 133Cs target with 27.5 MeV proton beams, and the straight-forward resin-based radiochemical separation, yielding 131Ba with high radionuclide purity. An average amount of 190 MBq of 131Ba was produced per irradiation. Apart from 0.1% isotopic impurity of 133Ba, no more side-products were detectable. For the first time, radiolabeling of the complexing agent macropa (known to be an appropriate 225Ac chelator) with 131Ba was reported and mild labeling conditions as well as reaction control using TLC systems were applicable. The radiopharmacological characterization of 131Ba-labeled macropa was carried out in healthy mice using uncomplexed [131Ba]Ba2+ as a reference, including biodistribution studies and small animal SPECT/CT. The results revealed the rapid bone uptake of free [131Ba]Ba2+ ions, whereas 131Ba-labeled macropa showed a fast renal clearance and significantly lower (P < 0.001) accumulation in the bones. We therefore conclude, that 131Ba is a promising “new” radionuclide for SPECT imaging purposes and delivers appropriate quality for preclinical investigations. Moreover, the successful labeling of macropa and the in vivo stability of the 131Ba-complex are viewed as a promising starting point for the development of new heavy earth alkaline metal chelators, especially for the therapeutically relevant radium isotopes. This enables 131Ba to achieve its goal as diagnostic match and monitoring tool for 223/224Ra.

Barium-131 is a promising SPECT-compatible radionuclide for nuclear medicine and a promising diagnostic match for the alpha emitters radium-223/-224 by providing similar chemical properties as well as physical half-lives. Methods: Herein, we report on the sufficient production route 133Cs(p,3n)131Ba by using 28 MeV proton beams. Moreover, a sufficient purification process, based on SR Resin, was applied. For the first time, radiolabeling of macropa (literature-known chelator) with barium-131 was performed. Biodistribution studies and small animal SPECT/CT measurements were carried out with [131Ba]Ba(NO3)2 as reference and 131Ba-labeled macropa. Results: An average of 190 MBq barium-131 per irradiation was obtained. The purification process led to barium-131 in high radiochemical purity. Only an isotopic impurity of 0.01% barium-133 was detectable. Radiolabeling methods under mild conditions and reaction controls based on TLC systems were successfully applied for the labeling of the chelator macropa. For the first time, small animal SPECT imaging was performed using [131Ba]Ba(NO3)2 and 131Ba-labeled macropa in healthy mice. Biodistribution studies revealed the expected rapid bone uptake of [131Ba]Ba2+ ions, whereas 131Ba-labeled macropa showed a fast clearance from the blood, thereby showing a significantly (P < 0.001) lower accumulation in the bone. Conclusion: Barium-131 is a promising SPECT radionuclide and delivers appropriate imaging qualities in small animals. Furthermore, the relative stability of the 131Ba-labeled macropa complex in vivo forms the basis for the development of sufficient new chelators for heavy alkaline earth metal ions, especially for radium isotopes. Those radionuclides and the necessary stable chelation are of great interest in the research field of targeted alpha therapy. Thereby, barium

Die Erfindung betrifft eine Verbindung, die - eine Calixaren-Einheit, die n Phenoleinheiten aufweist, wobei n 4, 5, 6 oder 8 ist; - eine Ethereinheit, die unter Ausbildung eines Kronenethers an die Calixaren-Einheit gebunden ist; und - zumindest eine Sulfonsäureamid-Einheit der Formel -O-(CH)ₚ-C(O)-NH-S(O)₂-R¹ aufweist, wobei die zumindest eine Sulfonsäureamid-Einheiten jeweils an die Calixaren-Einheit gebunden ist und R¹ jeweils aus der Gruppe ausgewählt ist, die aus einer perfluorierten verzweigten oder unverzweigten C₂-C₈-Alkylgruppe, einer perfluorierten Arylgruppe, und einer Gruppe Ar besteht, p eine Ganzzahl von 1 bis 4 ist und Ar eine Phenylgruppe ist, die mit einer oder mehreren perfluorierten verzweigten oder unverzweigten C₁-C₈-Alkylgruppen substituiert ist.

The radionuclides radium-223 and radium-224 are two alpha-emitting radionuclides with suitable properties for the TAT. To this date, radium-223 in form of [223Ra]radium chloride (Xofigo) is the only EMA and FDA approved alpha-emitting radiopharmaceutical. Due to its calcimimetic behavior, the radium ion is a bone-seeking therapeutic. To extend the radiopharmaceutical potential of both radionuclides, novel carrier systems have to be developed. Therefore, it is appropriate to investigate different kinds of nanoparticles for their ability to transport radium. Especially, a barium sulfate matrix seems to be sufficient since the principle of co-precipitating the sulfates of radium and barium allows an easy and fast synthesis of radium-doped nanoparticles. Beyond the incorporation of alpha-emitting radionuclides like radium-223 and radium-224, the homologue radionuclide barium-131 can be incorporated as well. Barium 131 decays by electron capture and provides suitable properties for diagnostic applications in nuclear medicine. Radium-223/-224 and barium-131 form a matched pair for new theragnostic approaches. In our research group, we are developing simple methods for the synthesis of small radiolabeled radium/barium sulfate nanoparticles. Furthermore, we are investigating suitable surface functionalizations to attach biological targeting moieties.

Therapieansätze in der Nuklearmedizin mit Radium-223 beschränken sich zurzeit auf die Behandlung von Knochenmetastasen. Ziel ist es, das Anwendungsfeld für Radium-223 zu erweitern. Dafür werden Liganden entwickelt, die Radium-223 stabil binden und gleichzeitig eine spezifische Erkennungssequenz für diverse Krebsentitäten besitzen. Calixkronen mit funktionalisierten Seitenketten sind dafür vielversprechend. Calix[4]krone-6-derivate mit Trifluormethylsulfonyl- [1] und 4-Nitrophenylsulfonyl-Seitenketten wurden synthetisiert und mittels NMR und UV/Vis auf ihre Komplexierungseigenschaften für Ba2+ untersucht. Anschließend wurden mittels eines Extraktionsassays Komplexierungen durchgeführt und die resultierenden Komplexe auf ihre Stabilität in Gegenwart anderer Ionen wie Ca2+ untersucht. Die Extraktionsversuche wurden im Chloroform/Wasser-Zweiphasensystem durchgeführt. Dabei wurde eine wässrige Metallsalzlösung ([133Ba]Ba2+ c.a. bzw. [224Ra]Ra2+ n.c.a.) definierter Konzentration vorgelegt und mit einer definierten Stoffmenge des jeweiligen Liganden in Chloroform für 1 h bei RT extrahiert. Die beiden Calixarenderivate wurden in einer vierstufigen Synthese hergestellt. Anschließende Komplexierungsversuche mit Ba(ClO4)2 verliefen erfolgreich und die Komplexe wurden mittels NMR-Spektroskopie nachgewiesen. Für beide Liganden wurden Stabilitätskonstanten von 5,3-6,3 (Ba2+) bzw. 5,8 (Ra2+) berechnet. Die Radiumkomplexe beider Liganden waren gegenüber Ca2+-Ionen stabil (Release <5%), der Bariumkomplex des Nitrophenylsulfonylliganden war ebenfalls stabil (Release <5%), der des Trifluormethylsulfonylliganden zeigte 30% Release. Die Wahl der sterisch anspruchsvollen und elektronenziehenden Sulfonylgruppen hatte positive Auswirkungen sowohl auf das Komplexierungsverhalten mit Ba2+ und Ra2+ als auch auf die Stabilität der resultierenden Komplex

1. Zielstellung: Durch die Alterung der Bevölkerung ist eine stetige Zunahme an Tumorneuerkrankungen zu verzeichnen und die Therapieoptimierung daher unabdingbar. Ein diskutierter Ansatz ist die Therapie mit Alphaemittern. Radionuklide wie 223/224Ra sind in der Lage, Tumorgewebe aufgrund ihres hohen linearen Energietransfers infolge einer Kaskade von Alpha-Zerfällen effizient zu zerstören ohne Beinflussung des gesunden Gewebes. Die stabile Fixierung von Barium- und Radiumionen kann durch Co-Fällung radiomarkierter [133Ba223/224Ra]Ba(Ra)SO4 Nanopartikel (NP) erfolgen, welche mit Alendronat funktionalisiert und anschließend mit einer targetspezifischen Einheit modifiziert werden. 2. Methodik Die Synthese der BaSO4-NP wurde durch Fällung realisiert. Die Partikelgröße wurde in Abhängigkeit der Reaktionsparameter (Lösungsmittelsystem, Verhältnis der Reaktanden, Fließgeschwindigkeit bei der Zugabe des zweiten Reaktanden) untersucht. NP-Größenverteilungen wurden mittels Dynamic Light Scattering (DLS) ermittelt. Radiomarkierungen wurden unter analogen Bedingungen unter Zugabe von [133Ba]Ba2+ und [224Ra]Ra2+ durchgeführt. 224Ra wurde als [224Ra]Ra(NO3)2 aus einer 228Th-Quelle durch Ionenaustauschchromatographie separiert. 133Ba wurde als [133Ba]BaCl2 kommerziell bezogen. 3. Ergebnisse Nach Auswertung aller Syntheseansätze wurde die Fällung der Partikel mit einem sechsfachen Bariumüberschuss in einer Ethanol/Wasser-Mischung (1/11) bei Raumtemperatur als beste Methode identifiziert. Die Einlagerung von [133Ba]Ba2+- und [224Ra]Ra2+-Ionen konnte nachgewiesen werden. Die radiomarkierten NP zeigten über 7 Tage keinen relevanten Release. Die Modifikation der Partikel mit Alendronat konnte mittels IR-Spektroskopie, EDXS-Analyse und UV/Vis-Spektroskopie na

Aim/Introduction: The Al18F-labeling method is as modern technique an alternative to conventional 18F-labeling procedures that allows radiofluorination of biomolecules such as peptides and proteins in a one-step procedure in aqueous solution [1]. In search for versatile applicable chelators, which allow stable binding of both 18F and radiometals such as 68Ga or 111In, a cyclohexanediamine-triazole-chelator was designed. This chelator was conjugated via copper-catalyzed azide-alkyne cycloaddition (CuAAC) to the well-known binding motif (glutamate-urea-lysine) of the prostate-specific membrane antigen (PSMA) [2-5] complemented by 2-azidoacetyl moiety as linker unit (ligand L1). Furthermore, ligand L2 was synthesized bearing a 6-azidohexanoyl moiety as linker to investigate the influence of the linker on the stability of final 18F or radiometal complex. The aim of this study was to investigate the radiopharmacological potential of L1 and L2 after radiolabeling regarding binding properties, cell internalization, and in vivo behavior in a murine prostate cancer model. Methods: For the in vitro assays PSMA-positive LNCaP cells were used. The incubation with the respective radiolabeled ligand (RCY>95%) was terminated via a cell harvester. Internalization experiments were carried out by the “acid wash” method. In vivo studies (biodistribution and small animal PET) were performed with mice bearing a prostate tumor. Results: In competition assays versus [177Lu]Lu-PSMA-617 (“gold standard”), the affinity of non-labeled L1 and L2 was slightly lower than that of PSMA-617. Saturation analysis of [68Ga]Ga-L1, [111In]In-L1, and [18F]F-L1 binding on LNCaP homogenate was comparable to [18F]F-L2 binding. The obtained Kd values were in a range of 20 to 30 nM. Internalization experiments with LNCaP cells revealed a lower uptake of the differently labeled L1 and L2 conjugates compared to [64Cu]Cu-PSMA-617. Furthermore, in vivo behavior of both [18F]F-L1 and [18F]F-L2 was investigated in

Understanding the coordination chemistry of heavy group 2 metals, especially of barium as surrogate for radium, is mandatory not only for radiopharmaceutical applications of radium. This is from high importance since radium-223 is the only approved therapeutic alpha-emitter (by EMA and FDA). Unfortunately, the applications are limited. To date, radium-223 is only in use as RaCl2 for the treatment of bone cancer metastases. To overcome this limitation, which is also true for other group 2 metals, special cage-like compounds have to be developed as ligands like sulfonated calix[4]crowns to stably bind the Ba2+ and Ra2+ to avoid a release in vivo. This will be the basis for a future application of heavy group 2 metals and not only of radium to treat other cancer entities than bone metastases. Ra2+ can then be included in radiopharmaceuticals which contain a chelator and a biologically active molecule part to find the tumor cell. For this purpose, a series of modified calix[4]crown-6 derivatives was synthesized to chelate barium, which serves as non-radioactive surrogate for radium-223/-224. These calixcrowns were functionalized sulfonate moieties including deprotonable groups and the corresponding barium complexes were synthesized. Stability constants of these complexes were measured using UV/Vis titration experiments to determine logK values. Further extraction studies were performed with [133Ba]Ba2+ and [224Ra]Ra2+ to further characterize the binding affinity of calixcrowns.

Durch fortschreitende Alterung der Bevölkerung ist eine kontinuierliche Zunahme von Tumorerkrankungen zu verzeichnen. Die Optimierung bestehender sowie die Entwicklung neuer Therapiekonzepte ist daher unabdingbar. Ein aktuell intensiv diskutierter Ansatz ist die nuklearmedizinische Therapie mit Alphastrahler-markierten Radiopharmaka. Radionuklide wie 223/224Ra oder 225Ac sind in der Lage, Tumorgewebe aufgrund ihres vergleichsweise hohen, linearen Energietransfers infolge einer Kaskade von Alpha-Zerfällen effizient zu zerstören. Ein theragnostischer Ansatz könnte mit dem „Matched Pair“ 131Ba/223/224Ra verfolgt werden. Die Herausforderung der stabilen Fixierung von Barium- und Radiumionen könnte durch Kofällung radiomarkierter [131/133Ba/223/224Ra]Ba(Ra)SO4-Nanopartikel (NP) erfolgen.

Ziel: Nach dem heutigen Stand sind keine geeigneten Chelatoren zur stabilen Komplexierung von schweren Erdalkalimetallen Barium und Radium bekannt. Da die Alphaemitter Radium-223 Radium-224 jedoch hohes therapeutisches Potential besitzen, ist die Entwicklung von multimodalen Liganden von großem Interesse. Ein solcher Ligand könnte eine Matched-Pair-Strategie eröffnen. Während Radium-223 und Radium-224 therapeutisch einsetzbar sind, besitzt Barium-131, welches analoge chemische Eigenschaften aufzeigt, gute diagnostische Eigenschaften. Bis jetzt ist die klinische Nutzung von Radium auf die Behandlung von Knochenmetastasen durch das calcimimetische Radiopharmakon [223Ra]RaCl2 (Xofigo®) beschränkt. Diese Anwendung gilt es zu erweitern. Methodik: Radium-224 wurde in Form seines Nitrats mittels Ionenaustauschchromatographie aus einem Thorium-228-Generator gewonnen. Als Bariumisotop für Extraktionsstudien wurde das langlebige und kommerziell erhältliche Radionuklid Barium-133 genutzt. Die Radiomarkierungen wurden mittels HPLC und Extraktionsstudien charakterisiert. Stabilitätskonstanten wurden mit stabilem Barium mittels UV/Vis und Kalorimetrie berechnet. Mit den Radiometallen Barium-133 und Radium-224 konnten vergleichbare Konstanten mittels Zwei-Phasen-Extraktion (Wasser/Chloroform) bestimmt werden. Als Liganden dienten unter anderem verschieden funktionalisierte, 18-Krone-6-überbrückte Calix[4]-arene. Ergebnisse: Es wurden vier Liganden auf Calix[4]-aren-Basis dargestellt und deren Interaktion mit Bariumionen untersucht. Ein relativ hohe Stabilitätskonstante konnte vor allem für ein Sulfonamid-Derivat ermittelt werden. Angeschlossene Extraktionsstudien zeigten für diesen Liganden eine nahezu vollständige Extraktion von [133Ba]Ba2+ aus der wässrigen Phase unter physiologischen Bedingungen. Analoge Versuche wurden mit [224Ra]Ra2+ durchgeführt. Der Komplex zeigte dabei eine ähnliche Stabilität. Schlussfolger

Radium is the heaviest known member of the alkaline earth metals and all 33 of its isotopes are radioactive. Two of these, radium-223 and radium-224, have suitable half-lives with 11.4 d and 3.6 d, respectively, and nuclear decay properties that make them useful tools for alpha particle therapy. Unfortunately, no suitable chelating agents are available for a stable complexation. Thus, radium-223 is only in use as radium chloride to treat bone metastases. For this purpose, a series of modified calix[4]crown-6 derivatives was synthesized to chelate heavy group 2 metal ions like barium, which serves as a non-radioactive surrogate for radium-223/-224. The calix[4]arene framework can be seen as an ideal platform to build an optimized chelator. Two of the four hydroxy groups of the lower rim can be functionalized as deprotonizable groups to form a neutral complex with barium or radium; the remaining two are bridged by a crown ether moiety. With this concept, the advantages of the electrostatic, macrocyclic, and cryptate effect are combined. Another benefit of the calixcrowns is their easy access. As a result, our calix[4]crowns were functionalized with either cyclic amide moieties or with deprotonizable groups like carboxylic acids or hydroxyl amines, and the corresponding barium complexes were synthesized. To prove the ability of these chelators for a further usage in radiopharmacy, stability constants of the corresponding barium complexes were determined by using NMR and UV/Vis titration to determine logK values. Further extraction studies were performed to characterize the binding affinity of calixcrowns to barium-133 and radium-224.

64Cu wird für die nuklearmedizinische Anwendung diskutiert. Neben der Kernumwandlung über Elektroneneinfang werden auch Positronen und Auger-Elektronen (AE) emittiert. Um deren Wirkungspotential an DNA zu untersuchen, wurden zunächst zwei Pyrenderivate mit unterschiedlicher Spacerlänge zwischen Pyreneinheit und DOTA-Chelator mit [64Cu]CuCl2 radiomarkiert. Die induzierten Strahlenschäden an pUC19 Plasmid-DNA wurden in Abhängigkeit von der Dosis untersucht.

A series of modified calix[4]crown-6 derivatives was synthesized to chelate the heavy group 2 metal barium, which serves as non-radioactive surrogate for radium-223/-224; radionuclides with promising properties for radiopharmaceutical use. These calixcrowns were functionalized either with cyclic amide moieties or proton-ionizable groups and the corresponding barium complexes were synthesized. Stability constants of these complexes were measured using NMR and UV/Vis titration techniques to determine logK values between 4.1 and 6.4. Further extraction studies were performed to characterize the binding affinity of calixcrowns to radioactive barium-133. Additionally, the ligands containing cyclic amides were investigated regarding their barriers for rotation using temperature dependent NMR measurements.

Die vorliegende Arbeit beschäftigt sich mit zwei Schwerpunkten. Zum einen sollen eine Vielzahl von Derivaten eines modifizierten Calix[4]arengründgerüsts sowie ein modifiziertes Calix[6]arenderivat bezüglich ihrer komplexbildenden Eigenschaften untersucht und Komplexbildungskonstanten mit einer Methode unter der Verwendung der radioaktiven Barium- und Radiumisotope ermittelt werden. Diese Methode ist zunächst zu entwickeln und die erhaltenen Ergebnisse mit den Berechnungsmethoden, welche innerhalb der Arbeitsgruppe etabliert und bezüglich der Calix[4]arenderivate anwendbar sind, zu vergleichen. Der Komplex mit der größten Stabilität soll festgestellt werden, um ihn weiter zu modifizieren oder bei ausreichender Stabilität erste biologische Studien beginnen zu können. Zum anderen soll auf der strukturellen Grundlage von Carbazol ein zusätzlicher makrocyclischer potentieller Komplexbildner in Form eines Calix[3]carbazolderivates synthetisiert werden, welcher alternativ zu den Calix[4]aren- und Calix[6]arenderivaten verwendet werden kann. Abschließend sollen die Komplexstabiltäten der Calix[n]arene, der synthetisierten Calix[3]carbazolderivate und kommerzieller Chelatoren mit den zweifach positiv geladenen Barium- und Radiumionen untereinander verglichen werden. Da es kein natürlich vorkommendes, nicht radioaktives Radiumisotop gibt, wird aufgrund seiner chemischen Ähnlichkeit stabiles Barium als Surrogat verwendet, um nicht radioaktive Untersuchungen durchführen zu können. Die erhaltenen Erkenntnisse sollen anschließend auf Untersuchungen mit Radium übertragen werden. Auch für die radioaktiven Studien wird zunächst das radioaktive Bariumisotop Barium-133 verwendet, da es eine längere Halbwertszeit besitzt als die verfügbaren Radiumisotope.