RESUMO
Mesocyclic chelating agents such as AAZTA and its derivatives have been recently reported to overcome the relatively low thermodynamic stability of metal complexes of acyclic chelating agents and the slow complexation kinetics of macrocyclic chelating agents. This work reports the preparation of a spirobicyclic hexadentate AAZTA-like chelating agent (TRASUTA) and the investigation of the thermodynamic, kinetic, and structural properties of the corresponding chelates with the PET-relevant Ga3+ and selected metal ions. A combination of analytical techniques allowed identification of a coordination isomerization process, involving the coordinating side arms and the inversion of a nitrogen atom and leading to lower thermodynamic and kinetic inertness with respect to mononuclear mesocyclic analogues. The bicyclic system of TRASUTA retains significant dynamics despite the conformational constraint imposed by the spiro-fusion, resulting in a lower stability of the corresponding metal chelates.
RESUMO
Optical imaging (OI) is gaining increasing attention in medicine as a non-invasive diagnostic imaging technology and as a useful tool for image-guided surgery. OI exploits the light emitted in the near-infrared region by fluorescent molecules able to penetrate living tissues. Cyanines are an important class of fluorescent molecules and by their conjugation to peptides it is possible to achieve optical imaging of tumours by selective targeting. We report here the improvements obtained in the synthesis of DA364, a small fluorescent probe (1.5 kDa) prepared by conjugation of pentamethine cyanine Cy5.5 to an RGD peptidomimetic, which can target tumour cells overexpressing integrin αvß3 receptors.
Assuntos
Integrina alfaVbeta3 , Integrina beta3 , Linhagem Celular Tumoral , Oligopeptídeos/químicaRESUMO
Two structurally constrained chelators based on a fused bicyclic scaffold, 4-amino-4-methylperhydro-pyrido[1,2-a][1,4]diazepin-N,N',N'-triacetic acids [(4R*,10aS*)-PIDAZTA (L1) and (4R*,10aR*)-PIDAZTA (L2)], were designed for the preparation of GaIII -based radiopharmaceuticals. The stereochemistry of the ligand scaffold has a deep impact on the properties of the complexes, with unexpected [Ga(L2)OH] species being superior in terms of both thermodynamic stability and inertness. This peculiar behavior was rationalized on the basis of molecular modeling and appears to be related to a better fit in size of GaIII into the cavity of L2. Fast and efficient formation of the GaIII chelates at room temperature was observed at pH values between 7 and 8, which enables 68 Ga radiolabeling under truly physiological conditions (pHâ 7.4).
Assuntos
Compostos Bicíclicos com Pontes/química , Quelantes/química , Complexos de Coordenação/química , Complexos de Coordenação/síntese química , Complexos de Coordenação/metabolismo , Cristalografia por Raios X , Teoria da Densidade Funcional , Radioisótopos de Gálio/química , Meia-Vida , Humanos , Concentração de Íons de Hidrogênio , Cinética , Conformação Molecular , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/metabolismo , Transferrina/químicaRESUMO
The relaxivity of Gd(HP-DO3A) was studied as a function of pH and buffer composition in order to identify the main factors of the observed relaxation enhancement due to the exchange of the coordinated hydroxyl proton. It was established that the paramagnetic relaxation time, T1M, of the coordinated hydroxyl proton is about 50% shorter than that of the protons in the coordinated water molecule. The control of the p K of the coordinated alcoholic -OH moiety in the ligand is fundamental to utilize the proton exchange enhanced relaxivity under physio/pathologic conditions. A new derivative of Gd(HP-DO3A) was synthesized by replacing the -CH3 group with a -CF3 moiety. In this complex, the -OH group becomes more acidic. Consequently, the maximum contribution of the proton exchange to the relaxivity is shifted to a lower pH region with the fluorinated ligand.
Assuntos
Meios de Contraste/química , Gadolínio/química , Compostos Heterocíclicos com 1 Anel/química , Imageamento por Ressonância Magnética , Compostos Organometálicos/química , Prótons , Meios de Contraste/síntese química , Concentração de Íons de Hidrogênio , Estrutura Molecular , Compostos Organometálicos/síntese químicaRESUMO
PURPOSE: To dissect the contributions to the longitudinal relaxivity (r1 ) of two commercial contrast agents (CAs), Gd-DOTA and Gd-HP-DO3A, and to synthesize/characterize a novel macrocyclic agent (Gd-Phen-DO3A) having superior r1 . METHODS: Longitudinal relaxation rates R1 of the CAs in saline with/without human serum albumin (HSA), ionized simulated body fluid (i-SBF), viscous simulated body fluid (v-SBF), and human plasma were measured. Results have been interpreted to evince the main determinants to the observed r1 values. RESULTS: In v-SBF or in the presence of HSA, r1 is enhanced for all complexes, reflecting the viscosity increase and a weak interaction with proteins. The CAs further differentiate in plasma, with a relaxivity increase (versus saline) of approximately 1, 1.5, and 2.5 mM-1 s-1 for Gd-DOTA, Gd-HPDO3A, and Gd-Phen-DO3A, respectively. R1 versus pH curves in i-SBF indicates that prototropic exchange sizably contributes to the relaxivity of Gd-HP-DO3A and Gd-Phen-DO3A. CONCLUSION: The major contributions to r1 in the physiological environment have been highlighted, namely, increased viscosity, complex-protein interaction, and prototropic exchange. The control of these terms allows the design of novel macrocyclic structures with enhanced r1 as a result of an improved interaction with plasma's macromolecules and the shift of the prototropic exchange to physiological pH. Magn Reson Med 78:1523-1532, 2017. © 2016 International Society for Magnetic Resonance in Medicine.
Assuntos
Meios de Contraste/química , Compostos Heterocíclicos/química , Imageamento por Ressonância Magnética/métodos , Compostos Organometálicos/química , Meios de Contraste/análise , Meios de Contraste/metabolismo , Compostos Heterocíclicos/sangue , Compostos Heterocíclicos/metabolismo , Humanos , Modelos Biológicos , Compostos Organometálicos/sangue , Compostos Organometálicos/metabolismo , Ligação Proteica , Albumina Sérica/química , Albumina Sérica/metabolismo , ViscosidadeRESUMO
Bifunctional chelating agents (BFCAs) combine the complexing properties of a multidentate ligand with the presence of a free reactive functional group, mainly devoted to conjugation purposes. Indeed, products obtained by conjugation of a BFCA to a biomolecule and coordination of a suitable metal ion are widely applied in medicine nowadays as diagnostic and therapeutic agents. BFCAs are generally prepared through multi-step syntheses and with extensive application of protection-deprotection strategies, due to the large number of functional groups involved. Hydrolytic enzymes, with their unique chemoselectivity, provided the best results in the preparation of three different BFCAs based on very useful and well known ligand platforms.
Assuntos
Quelantes/química , Animais , Aspergillus oryzae/enzimologia , Soluções Tampão , Candida/enzimologia , Quelantes/síntese química , Enzimas/química , Hidrólise , Hidróxidos/química , Cinética , Ligantes , Lipase/química , Solventes/química , Suínos , TermodinâmicaRESUMO
An unexpected nitro group displacement during a nitro-Mannich reaction led to the one-pot formation of the octahydro-2H-pyrazino[1,2-a]pyrazine core, representing the shortest access to date to this pharmacologically relevant heterobicyclic system. A mechanistic hypothesis is suggested and supported by specific experiments and HRMS analysis of reaction mixtures.
RESUMO
Bifunctional chelating agents (BFCAs) are molecules which contain two different moieties: a strong metal chelating unit and a reactive functional group. The latter is directed to react with amines, thiols, alcohols or other reactive molecules to form stable covalent bonds while the chelating moiety is able to strongly coordinate a metal ion. In this way, it is possible to label a molecule of interest (e.g. an antibody or a peptide) with a metal or a radioactive metal ion. Of all the ligands reported so far, those based on a polyamino polycarboxylic structure are most efficient and are widely employed for the chelation of metal ions. The resulting metal complexes have found a broad range of applications in chemistry, biology and medicine. Diagnostic imaging (MRI, SPECT, PET), molecular imaging, tumour therapy and luminescent materials are only a few examples. The present critical review gives an overview of the syntheses and most important applications of polyamino polycarboxylic BFCAs (334 references).
Assuntos
Quelantes/síntese química , Acetatos/química , Ácidos Acíclicos/química , Quelantes/química , Compostos Heterocíclicos/química , Ácido Pentético/químicaRESUMO
Polyaminopolycarboxylic acids are a well known class of ligands employed for metal ion complexation. Despite the large commercial availability, reports of their use as substrates for direct structural modifications are rare. Herein we report a simple and efficient protocol for the preparation of substituted polyaminopolycarboxylic ligands relying on a one-pot N-alkylation-Stevens rearrangement cascade.
Assuntos
Ácidos Carboxílicos/química , Poliaminas/química , Alquilação , Ligantes , Estrutura MolecularRESUMO
Eu(HP-DO3A) is present in solution as a mixture of two diastereoisomers whose alcoholic groups are the source of the mobile protons for the CEST effect. The exchange is base catalyzed. Two novel EuIII complexes of HP-DO3A-like ligands containing an amino or a carboxylate functionality in the proximity of the -OH groups showed the occurrence of intramolecular catalysis of the prototropic exchange. New insights into the role of the intramolecular proton exchange on the CEST properties have been gained.
RESUMO
The set-up of reversible binding interactions between the hydrophobic region of macrocyclic GBCAs (Gadolinium Based Contrast Agents) and SO3 -/OH containing pyrene derivatives provides new insights for pursuing relaxivity enhancements of this class of MRI contrast agents. The strong binding affinity allows attaining relaxation enhancements up to 50% at pyrene/GBCA ratios of 3 : 1. High resolution NMR spectra of the Yb-HPDO3A/pyrene system fully support the formation of a supramolecular adduct based on the set-up of hydrophobic interactions. The relaxation enhancement may be accounted for in terms of the increase of the molecular reorientation time (τ R) and the number of second sphere water molecules. This effect is maintained in blood serum and in vivo, as shown by the enhancement of contrast in T 1w-MR images obtained by simultaneous injection of GBCA and pyrene derivatives in mice.
RESUMO
Complexes of Gd, Eu and Yb(III) have been prepared with a series of heptadentate ligands related to the parent complex AAZTA, based on the 6-methyl-6-aminoperhydrodiazepine moiety. For (RR) and (RS)-diastereoisomers of a di-glutarate ligand, solution NMR studies revealed the presence of two major species that undergo water exchange rates at Gd differing by a factor of six. Comparison of solution hydration states for Eu(III) complexes reveals that each complex possesses two bound water molecules. The absence of a good correlation of (1)H NMR pseudo-contact shifts for Eu and Yb analogues is suggested to arise from a change in hydration state between Eu and Yb.
Assuntos
Azepinas/química , Elementos da Série dos Lantanídeos/química , Compostos Organometálicos/química , Termodinâmica , Água/química , Ligantes , Estrutura Molecular , EstereoisomerismoRESUMO
Two novel bifunctional chelating agents (BFCAs) based on the structure of 10-(2-hydroxypropyl)-1,4,7-tetraazacyclododecane-1,4,7-triacetic acid (HP-DO3A) have been designed and synthesized. BFCAs are able to strongly coordinate metals (e.g. Gd(iii)) and radiometals (e.g.(90)Y(iii), (177)Lu(iii) and (111)In(iii)), and find applications in the synthesis of products for the imaging and treatment of several cancer types. Indeed, these two BFCAs have been employed in solid-phase peptide synthesis (SPPS) and coupled to well-known peptides such as bombesin and RGD analogues in order to target tumor cells. We also report here the conjugation of one of them to Lys(8)-oxytocin and radiolabeling with (111)In(iii) to obtain a new radiopharmaceutical product with potential applications in the diagnosis of tumors over-expressing oxytocin receptors.
Assuntos
Quelantes/química , Quelantes/síntese química , Compostos Heterocíclicos com 1 Anel/química , Compostos Heterocíclicos com 1 Anel/síntese química , Radioisótopos de Índio/química , Marcação por Isótopo , Ligantes , Ocitocina/química , Peptídeos/químicaRESUMO
A dinuclear gadolinium(III) chelate containing two moieties of diethylenetriaminepentaacetic acid (DTPA), covalently conjugated to an analogue of deoxycholic acid, was synthesized and thoroughly characterized. A full relaxometric analysis was carried out, consisting of 1)â the acquisition of nuclear magnetic resonance dispersion (NMRD) profiles in various media; 2)â the study of binding affinity to serum albumin; 3)â the measurement of 17 O transverse relaxation rate versus temperature, and 4)â a transmetallation assay. In vivo biodistribution MRI studies at 1â T and blood pharmacokinetics assays were carried out in comparison with Gd-DTPA (Magnevist) and gadocoletic acid trisodium salt (B22956/1), two well-known Gd complexes that share the same chelating cage and the same deoxycholic acid residue of the Gd complex investigated herein ((GdDTPA)2 -Chol). High affinity for plasma protein and, in particular, the availability of more than one binding site, allows the complex to reach a fairly high relaxivity value in plasma (â¼20â mm-1 s-1 , 20â MHz, 310â K) as well as to show unexpectedly enhanced properties of blood pooling, with an elimination half-life in rats approximately seven times longer than that of B22956/1.
Assuntos
Meios de Contraste/química , Meios de Contraste/farmacocinética , Ácido Desoxicólico/análogos & derivados , Ácido Desoxicólico/farmacocinética , Gadolínio DTPA/análogos & derivados , Gadolínio DTPA/farmacocinética , Animais , Meios de Contraste/síntese química , Meios de Contraste/metabolismo , Ácido Desoxicólico/síntese química , Ácido Desoxicólico/metabolismo , Feminino , Gadolínio DTPA/síntese química , Gadolínio DTPA/metabolismo , Imageamento por Ressonância Magnética , Masculino , Ratos , Ratos Wistar , Albumina Sérica/metabolismo , Distribuição TecidualRESUMO
The Gd(iii)-complexes of three novel HP-DO3A-like ligands have been investigated to assess the relationship between relaxometry and intramolecular catalysis of the proton exchange. The structures of these ligands differ from the parent HP-DO3A because the methyl group of the hydroxy-propyl arm has been replaced by -Ph-OH, -Ph-NH2 and -Ph-COOH, respectively. The phenol, amine and carboxylate functionalities display an intramolecular H-bonding with the coordinated hydroxyl moiety that affects either the pK values of the involved functionalities and the rate of the proton exchange process.
RESUMO
The glutamine transporting system is up-regulated in tumor cells because cell proliferation requires the uptake of large quantities of glutamine. It has been found that the paramagnetic magnetic resonance imaging (MRI) reporter Gd-DOTAMA-C6-Gln, where the glutamine residue is covalently bound to the Gd chelate through a C6 spacer, accumulates in tumor cells both "in vitro" and "in vivo" experiments. The observation that the relaxivity of cellular pellets does not increase with the increase in the amounts of entrapped Gd chelate is taken as an indication that the internalization has occurred through receptor mediated endocytosis. The iv administration of Gd-DOTAMA-C6-Gln allowed the MRI visualization of tumor masses in A/J mice grafted with the murine neuroblastoma cell line Neuro-2a and in Her-2/neu transgenic mice developing multiple mammary carcinoma, respectively.
Assuntos
Proteínas de Transporte/metabolismo , Meios de Contraste , Gadolínio , Glutamina/metabolismo , Neoplasias Experimentais/diagnóstico , Compostos Organometálicos/síntese química , Animais , Linhagem Celular Tumoral , Quelantes/síntese química , Meios de Contraste/farmacocinética , Feminino , Compostos Heterocíclicos com 1 Anel/química , Humanos , Imageamento por Ressonância Magnética , Neoplasias Mamárias Experimentais/diagnóstico , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Transgênicos , Neoplasias Experimentais/patologia , Compostos Organometálicos/farmacocinética , Ratos , Receptor ErbB-2/genética , Transplante HeterólogoRESUMO
UNLABELLED: Gastrin-releasing peptide receptors (GRP-R) are upregulated in many cancers, including prostate, breast, and lung. We describe a new radiolabeled bombesin (BBN) analog for imaging and systemic radiotherapy that has improved pharmacokinetics (PK) and better retention of radioactivity in the tumor. METHODS: DO3A-CH2CO-G-4-aminobenzoyl-Q-W-A-V-G-H-L-M-NH2 (AMBA) was synthesized and radiolabeled. The human prostate cancer cell line PC-3 was used to determine the binding (Kd), retention, and efflux of 177Lu-AMBA. Receptor specificity was determined by in vitro autoradiography in human tissues. PK and radiotherapy studies were performed in PC-3 tumor-bearing male nude mice. RESULTS: 177Lu-AMBA has a high affinity for the GRP-R (Kd, 1.02 nmol/L), with a maximum binding capacity (Bmax) of 414 fmol/10(6) cells (2.5 x 10(5) GRP-R/cell). Internalization was similar for 177Lu-AMBA (76.8%), 177Lu-BBN8 (72.9%), and 125I-[Tyr4]-BBN (74.9%). Efflux was markedly lower for 177Lu-AMBA (2.9%) compared with 177Lu-BBN8 (15.9%) and 125I-[Tyr4]-BBN (46.1%). By receptor autoradiography, Lu-AMBA binds specifically to GRP-R (0.8 nmol/L) and to the neuromedin B receptor (NMB-R) (0.9 nmol/L), with no affinity for the bb3 receptor (>1,000 nmol/L). 177Lu-AMBA was renally excreted (55 %ID 1 h [percentage injected dose at 1 h]); tumor uptake at 1 and 24 h was 6.35 %ID/g and 3.39 %ID/g, respectively. One or 2 doses of 177Lu-AMBA (27.75 MBq/dose) significantly prolonged the life span of PC-3 tumor-bearing mice (P < 0.001 and P < 0.0001, respectively) and decreased PC-3 tumor growth rate over controls. When compared using World Health Organization criteria, mice receiving 2 doses versus 1 dose of 177Lu-AMBA demonstrated a shift away from stable/progressive disease toward complete/partial response; by RECIST (Response Evaluation Criteria in Solid Tumors), median survival increased by 36% and time to progression/progression-free survival increased by 65%. CONCLUSION: 177Lu-AMBA binds with nanomolar affinity to GRP-R and NMB-R, has low retention of radioactivity in kidney, demonstrates a very favorable risk-benefit profile, and is in phase I clinical trials.
Assuntos
Regulação da Expressão Gênica , Lutécio/farmacologia , Oligopeptídeos/farmacologia , Peptídeos/química , Neoplasias da Próstata/radioterapia , Radioisótopos/farmacologia , Receptores da Bombesina/biossíntese , Animais , Ligação Competitiva , Bombesina/química , Linhagem Celular Tumoral , Progressão da Doença , Humanos , Masculino , Camundongos , Camundongos Nus , Oligopeptídeos/química , Receptores da Bombesina/agonistasRESUMO
The synthesis of a new nonacoordinating ligand based on an AMPED (6-amino-6-methylperhydro-1,4-diazepine) scaffold functionalized by three picolinate (6-carboxy-2-methylpyridine) arms is described. Coordination of lanthanide cations (Ln = Eu and Tb) was investigated by spectrophotometric titrations monitored by UV-Vis absorption and steady-state emission spectroscopy, showing the formation of [LnL] complexes in aqueous solutions. The corresponding Eu and Tb complexes were isolated and characterized, and their spectroscopic properties (luminescence quantum yields, excited state lifetimes) were determined in buffered water (TRIS/HCl, pH 7.4) and compared to the data reported in the literature for related systems. DFT modelling of the complexes showed the picolinate arms to be perfectly wrapped around the Ln(3+) cations, affording an excellent shielding of the metal as confirmed by the determination of the hydration number of q = 0 for both complexes. The high resolution emission spectrum was used to determine the radiative lifetime of Eu in the complex (τrad = 3.05 ms) and the metal-centred luminescence quantum yield (0.20). The modest 0.10 overall luminescence quantum yield of the Eu complex is a consequence of an energy transfer with medium efficiency (0.50) and a low metal centred luminescence efficiency attributed in part to the presence of numerous NH and CH bonds in close proximity to the metal centre.
Assuntos
Azepinas/química , Quelantes/química , Elementos da Série dos Lantanídeos/química , Ácidos Picolínicos/química , Quelantes/síntese química , Ligantes , Espectrofotometria UltravioletaRESUMO
A series of structurally different Gd(III) conjugates incorporating a bile acid moiety have been prepared. Polyaminopolycarboxylic ligands such as diethylenetriaminepentaacetic acid (DTPA) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetracetic acid (DOTA) have been selected as chelating subunit for the Gd(III) ion. Cholic acid, cholylglycine, and cholyltaurine have been incorporated as the bile acid moieties. In first generation conjugates the Gd(III) complex is linked to the carboxyl group of cholic acid. Second generation conjugates feature the attachment of the Gd(III) complex to the 3 position of the steroidic backbone of the bile acid. Finally, in third generation conjugates the Gd(III) complex is attached to the epsilon nitrogen atom of cholyllysine. The conjugates are eliminated through the biliary route to a various extent (7.5 to 77% in rats) according to their structural features. Among the most promising terms, a second generation conjugate in which the Gd(III) complex is linked to cholic acid through the 3alpha hydroxy group seems to enter hepatocytes using the Na(+)/taurocholate transporter. Noticeably, some of the second generation conjugates are characterized by very high tolerabilities (LD(50) up to 9.5 mmol/kg) after intravenous administration in mice.
Assuntos
Ácidos e Sais Biliares/química , Quelantes/química , Meios de Contraste/síntese química , Gadolínio/química , Hepatócitos/metabolismo , Compostos Organometálicos/síntese química , Animais , Meios de Contraste/química , Meios de Contraste/farmacocinética , Feminino , Hemodinâmica/efeitos dos fármacos , Compostos Heterocíclicos com 1 Anel/química , Dose Letal Mediana , Imageamento por Ressonância Magnética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Compostos Organometálicos/química , Compostos Organometálicos/farmacocinética , Ácido Pentético/química , Coelhos , RatosRESUMO
We report the synthesis of novel chelates of Gd and (68)Ga with DPTA, DOTA, HP-DOA3, as well as with AAZTA, a novel chelating agent developed by our research group. These chelating agents were appropriately conjugated, prior to metal complexation, with DB58, an RGD peptidomimetic, conformationally constrained on an azabicycloalkane scaffold and endowed with high affinity for integrin α(ν)ß(3) . Because α(ν)ß(3) is involved in neo-angiogenesis in solid tumors and is also directly expressed in cancer cells (e.g. glioblastomas, melanomas) and ovarian, breast, and prostate cancers, these constructs could prove useful as molecular imaging probes in cancer diagnosis by MRI or PET techniques. Molecular modeling, integrin binding assays, and relaxivity assessments allowed the selection of compounds suitable for multiple expression on dendrimeric or nanoparticulate structures. These results also led us to an exploratory investigation of (68)Ga complexation for the promising (68)Ga-PET technique; the AAZTA complex 15((68)Ga) exhibited uptake in a xenograft model of glioblastoma, suggesting potentially useful developments with new probes with improved affinity.