RESUMEN
Thorium-232 (Th-232) is a promising fuel for advanced nuclear reactors. However, in case of internal human exposure to Th, there is currently no effective modality for its removal from liver and skeleton or for mitigating its effect. The FDA-approved agent, diethylenetriaminepentaacetate (DTPA), can remove Th and other actinides from blood circulation only. For the first time, a rationally-selected polyherbal hepatoprotective i.e. Liv52® (L52S), was evaluated in-combination with DTPA for its Th decorporation ability in Swiss mice. Inductively-coupled plasma mass spectroscopic analysis showed that oral administration of L52S in conjunction with DTPA significantly decreased Th burden from liver (20 %) and skeleton (33 %) as well as enhanced Th excretion (â¼2.5 folds) through urine in comparison to DTPA or L52S alone. The combinatorial therapy was found to be complementary in-action, ameliorating Th-induced tissue damage in liver, spleen, and bone more effectively than monotherapy. Furthermore, markers of liver function (alanine transaminase) and liver inflammation and fibrosis (NF-κB & keratin) further validated the beneficial effect of L52S. The human consumption of L52S for various liver disorders further supports its clinical application for Th decorporation and mitigation of its health effects.
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Hígado , Ácido Pentético , Torio , Animales , Torio/toxicidad , Ácido Pentético/química , Ratones , Hígado/efectos de los fármacos , Hígado/metabolismo , Extractos Vegetales/farmacología , Extractos Vegetales/química , Masculino , Quelantes/farmacología , Quelantes/química , Bazo/efectos de los fármacos , Bazo/metabolismoRESUMEN
ABSTRACT: Skin contamination by α-emitting actinides such as plutonium and americium is a risk for workers during nuclear fuel production and reactor decommissioning. Decontamination of skin is an important medical countermeasure to limit potential internal contamination, particularly in the case of injured skin. Current recommendations include undressing of the victim followed by skin washing using soap or chelating agents, such as diethylene triamine pentaacetic acid (DTPA). The goal of the present work is to assess the efficacy of a novel Ca-DTPA loaded gel to decontaminate injured skin exposed to plutonium or americium as compared to recommended treatments. For decontaminant testing on injured skin, whole body skin was obtained from euthanized rats and lesions created using a metallic brush. Delimited test areas were contaminated with plutonium or americium solutions of known properties. Various protocols were tested including time before contamination, duration of gel application, washing steps, as well as the concomitant addition or not of dressings. Activity was measured in each decontamination product and in skin. Data indicate that healthy skin was easier to decontaminate than damaged skin. On injured skin, we demonstrated an increased decontamination efficacy of the Ca-DTPA gel formulation as compared to the solution. Importantly, gel application alone was effective, and further gel applications could be used for residual activity.
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Americio , Descontaminación , Ácido Pentético , Plutonio , Piel , Animales , Descontaminación/métodos , Ratas , Ácido Pentético/química , Piel/lesiones , Piel/metabolismo , Piel/efectos de los fármacos , Quelantes/farmacología , Masculino , Ratas Sprague-Dawley , Geles/químicaRESUMEN
Prostate specific membrane antigen (PSMA) is an excellent target for imaging and treatment of prostate carcinoma (PCa). Unfortunately, not all PCa cells express PSMA. Therefore, alternative theranostic targets are required. The membrane protein prostate stem cell antigen (PSCA) is highly overexpressed in most primary prostate carcinoma (PCa) cells and in metastatic and hormone refractory tumor cells. Moreover, PSCA expression positively correlates with tumor progression. Therefore, it represents a potential alternative theranostic target suitable for imaging and/or radioimmunotherapy. In order to support this working hypothesis, we conjugated our previously described anti-PSCA monoclonal antibody (mAb) 7F5 with the bifunctional chelator CHX-Aâ³-DTPA and subsequently radiolabeled it with the theranostic radionuclide 177Lu. The resulting radiolabeled mAb ([177Lu]Lu-CHX-Aâ³-DTPA-7F5) was characterized both in vitro and in vivo. It showed a high radiochemical purity (>95%) and stability. The labelling did not affect its binding capability. Biodistribution studies showed a high specific tumor uptake compared to most non-targeted tissues in mice bearing PSCA-positive tumors. Accordingly, SPECT/CT images revealed a high tumor-to-background ratios from 16 h to 7 days after administration of [177Lu]Lu-CHX-Aâ³-DTPA-7F5. Consequently, [177Lu]Lu-CHX-Aâ³-DTPA-7F5 represents a promising candidate for imaging and in the future also for radioimmunotherapy.
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Carcinoma , Ácido Pentético , Animales , Ratones , Masculino , Ácido Pentético/química , Distribución Tisular , Próstata , Línea Celular Tumoral , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Monoclonales/química , Células Madre , Carcinoma/tratamiento farmacológico , Lutecio/químicaRESUMEN
PURPOSE: Epidermal growth factor receptors (EGFRs) are overexpressed in a wide range of tumors and are attractive candidates to target in targeted therapies. This study aimed to introduce a novel radiolabeled compound, 177Lu-cetuximab-PAMAM G4, for the treatment of EGFR-expressing tumors. METHODS: In this study, the cetuximab mAb was bound to PAMAM G4 and labeled with 177Lu via DTPA-CHX chelator. The synthesized nanosystem was confirmed by different analyses such as DLS, FT-IR, TEM, and RT-LC. Cell viability of the radioimmunoconjugate was assessed over the EGFR-expressing cell line of SW480. The biodistribution of 177Lu-Cetuximab-PAMAMG4 was determined in different intervals after injection of the radiolabeled compound in normal and tumoral nude mice via scarification and SPECT images. RESULTS: The average size of PAMAM G4 and PAMAM-Cetuximab-DTPA-CHX nanoparticles were 2 and 70 nm, respectively. 177Lu-Cetuximab-PAMAMG4 was prepared with radiochemical purity of more than 98%. The survival rates of SW480 cells at 24, 48, and 72 h post-treatment with177Lu-Cetuximab-PAMAMG4 (500 nM) were 18%, 15%, and 14%, respectively. The biodistribution studies showed a significant accumulation of 177Lu-Cetuximab-PAMAM in the EGFR-expressing tumor. CONCLUSION: According to the results, this new agent can be considered as an efficient therapeutic complex for tumors expressing EGFR receptors.
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Inmunoconjugados , Neoplasias , Animales , Ratones , Cetuximab , Medicina de Precisión , Inmunoconjugados/metabolismo , Distribución Tisular , Ratones Desnudos , Espectroscopía Infrarroja por Transformada de Fourier , Receptores ErbB/metabolismo , Ácido Pentético/química , Línea Celular TumoralRESUMEN
Considering the potential effect of the ambient temperature on soil microorganisms during heavy metal immobilization by hydrochar, 60 days of soil incubation was conducted to explore the impact of ambient temperature (5, 25, and 35 °C) on the immobilization of Pb and Zn by chitosan-magnetic sawdust hydrochar (CMSH) and magnetic chitosan hydrochar (MCH). The results showed that soil pH was relatively high and total organic carbon (TOC) was slightly lower in the 35 °C treatment. The diethylenetriaminepentaacetic acid (DTPA) available state content decreased significantly with the temperature increasing. Meanwhile, the ratios of stable Pb and Zn in the sequential extraction method proposed by the European Community Bureau of Reference (BCR) gradually increased with increasing temperature. The heatmap based on microbial community showed that elevated temperature not only favored the enrichment of metal-stable phyla, such as Chloroflexi, but was also involved in inhibiting the growth of Firmicutes, Actinobacteriota, and Proteobacteria. Meanwhile, different genera (Fonticella and Bacillus) in the Firmicutes phylum had distinct responses to temperature as well as to heavy metal immobilization effects. Subsequently, redundancy analysis confirmed that Chloroflexi and Fonticella were positively correlated with temperature and stable state metal content, while Actinobacteriota and Bacillus were negatively correlated with temperature and were positively correlated with DTPA available metal content. Moreover, Pb and Zn indicators displayed significant correlations for the dominant genera (R2 > 0.8, p < 0.02).
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Bacillus , Quitosano , Metales Pesados , Contaminantes del Suelo , Suelo/química , Plomo/análisis , Microbiología del Suelo , Temperatura , Metales Pesados/análisis , Bacterias , Firmicutes , Zinc/análisis , Ácido Pentético/química , Contaminantes del Suelo/análisisRESUMEN
Site-specifically modified radioimmunoconjugates exhibit superior in vitro and in vivo behavior compared to analogues synthesized via traditional stochastic methods. However, the development of approaches to site-specific bioconjugation that combine high levels of selectivity, simple reaction conditions, and clinical translatability remains a challenge. Herein, we describe a novel solution to this problem: the use of dual-variable domain immunoglobulins (DVD-IgG). More specifically, we report the synthesis, in vitro evaluation, and in vivo validation of a 177Lu-labeled radioimmunoconjugate based on HER2DVD, a DVD-IgG containing the HER2-targeting variable domains of trastuzumab and the catalytic variable domains of IgG h38C2. To this end, we first modified HER2DVD with a phenyloxadiazolyl methlysulfone-modified variant of the chelator CHX-Aâ³-DTPA (PODS-CHX-A''-DTPA) and verified the site-specificity of the conjugation for the reactive lysines within the catalytic domains via chemical assay, MALDI-ToF mass spectrometry, and SDS-PAGE. The chelator-bearing immunoconjugate was subsequently labeled with [177Lu]Lu3+ to produce the completed radioimmunoconjugate, [177Lu]Lu-CHX-Aâ³-DTPAPODS-HER2DVD, in >80% radiochemical conversion and a specific activity of 29.5 ± 7.1 GBq/µmol. [177Lu]Lu-CHX-Aâ³-DTPAPODS-HER2DVD did not form aggregates upon prolonged incubation in human serum, displayed 87% stability to demetalation over a 7 days of incubation in serum, and exhibited an immunoreactive fraction of 0.95 with HER2-coated beads. Finally, we compared the pharmacokinetic profile of [177Lu]Lu-CHX-Aâ³-DTPAPODS-HER2DVD to that of a 177Lu-labeled variant of trastuzumab in mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts. The in vivo performance of [177Lu]Lu-CHX-Aâ³-DTPAPODS-HER2DVD matched that of 177Lu-labeled trastuzumab, with the former producing a tumoral activity concentration of 34.1 ± 12.1 %ID/g at 168 h and tumor-to-blood, tumor-to-liver, and tumor-to-kidney activity concentration ratios of 10.5, 9.6, and 21.8, respectively, at the same time point. Importantly, the DVD-IgG did not exhibit a substantially longer serum half-life than the traditional IgG despite its significantly larger size (202 kDa for the former vs 148 kDa for the latter). Taken together, these data suggest that DVD-IgGs represent a viable platform for the future development of highly effective site-specifically labeled radioimmunoconjugates for diagnostic imaging, theranostic imaging, and radioimmunotherapy.
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Neoplasias de la Mama , Inmunoconjugados , Humanos , Animales , Ratones , Femenino , Inmunoconjugados/uso terapéutico , Línea Celular Tumoral , Trastuzumab/uso terapéutico , Trastuzumab/farmacocinética , Quelantes/química , Neoplasias de la Mama/tratamiento farmacológico , Ácido Pentético/química , Inmunoglobulina G/uso terapéuticoRESUMEN
Speciation of actinides, and more particularly bioligand-binding ability, influences in vivo behavior. Understanding these interactions is essential for estimation of radiological dose and improvement of decorporation strategies for accidentally contaminated victims. Because the handling of actinides imposes overwhelming difficulties, in vitro assays carried out in physiological conditions are lacking and data regarding such interactions are scarce. In this study, we used a bi-compartmental and dynamic assay, providing physiological conditions (presence of inorganic ions, pH, temperature) to explore interactions between the actinides plutonium (Pu) and americium (Am) and endogenous (proteins transferrin and ferritin) or exogenous ligands (the chelating agent diethylenetriaminpentaacetic acid, DTPA). In this assay, an agarose gel represents the retention compartment of actinides and a dynamic fluid phase, the transfer compartment. The proportion of actinides transferred from static to dynamic phase reflects interactions between Pu/Am and various ligands. The results show differences in the formation of actinide-protein or actinide-DTPA complexes in physiologically relevant media depending on which ligand is present and where. We observed differential behavior for Pu and Am similar to in vivo studies. Thus, our assay may be used to determine the ability of various actinides to interact with specific proteins or with drug candidates for decorporation in complex physiologically relevant environments.
Asunto(s)
Elementos de Series Actinoides , Plutonio , Ligandos , Elementos de Series Actinoides/química , Americio/análisis , Plutonio/química , Ácido Pentético/químicaRESUMEN
Neuroendocrine prostate cancer (NEPC) is a lethal subtype of prostate cancer with limited meaningful treatment options. NEPC lesions uniquely express delta-like ligand 3 (DLL3) on their cell surface. Taking advantage of DLL3 overexpression, we developed and evaluated lutetium-177 (177Lu)-labeled DLL3-targeting antibody SC16 (177Lu-DTPA-SC16) as a treatment for NEPC. SC16 was functionalized with DTPA-CHX-A" chelator and radiolabeled with 177Lu to produce 177Lu-DTPA-SC16. Specificity and selectivity of 177Lu-DTPA-SC16 were evaluated in vitro and in vivo using NCI-H660 (NEPC, DLL3-positive) and DU145 (adenocarcinoma, DLL3-negative) cells and xenografts. Dose-dependent treatment efficacy and specificity of 177Lu-DTPA-SC16 radionuclide therapy were evaluated in H660 and DU145 xenograft-bearing mice. Safety of the agent was assessed by monitoring hematologic parameters. 177Lu-DTPA-SC16 showed high tumor uptake and specificity in H660 xenografts, with minimal uptake in DU145 xenografts. At all three tested doses of 177Lu-DTPA-SC16 (4.63, 9.25, and 27.75 MBq/mouse), complete responses were observed in H660-bearing mice; 9.25 and 27.75 MBq/mouse doses were curative. Even the lowest tested dose proved curative in five (63%) of eight mice, and recurring tumors could be successfully re-treated at the same dose to achieve complete responses. In DU145 xenografts, 177Lu-DTPA-SC16 therapy did not inhibit tumor growth. Platelets and hematocrit transiently dropped, reaching nadir at 2 to 3 wk. This was out of range only in the highest-dose cohort and quickly recovered to normal range by week 4. Weight loss was observed only in the highest-dose cohort. Therefore, our data demonstrate that 177Lu-DTPA-SC16 is a potent and safe radioimmunotherapeutic agent for testing in humans with NEPC.
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Anticuerpos Monoclonales Humanizados , Carcinoma Neuroendocrino , Péptidos y Proteínas de Señalización Intracelular , Proteínas de la Membrana , Neoplasias de la Próstata , Radioinmunoterapia , Animales , Anticuerpos Monoclonales Humanizados/química , Anticuerpos Monoclonales Humanizados/uso terapéutico , Carcinoma Neuroendocrino/radioterapia , Quelantes/química , Humanos , Péptidos y Proteínas de Señalización Intracelular/antagonistas & inhibidores , Péptidos y Proteínas de Señalización Intracelular/inmunología , Ligandos , Lutecio , Masculino , Proteínas de la Membrana/antagonistas & inhibidores , Ratones , Ácido Pentético/química , Neoplasias de la Próstata/radioterapia , Radioisótopos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
This study aims to establish new labeling methods for no-carrier-added radio-Pt (191Pt) and to evaluate the in vitro properties of 191Pt-labeled agents compared with those of agents labeled with the common emitter 111In. 191Pt was complexed with the DNA-targeting dye Hoechst33258 via diethylenetriaminepentaacetic acid (DTPA) or the sulfur-containing amino acid cysteine (Cys). The intranuclear fractions of 191Pt- and 111In-labeled Hoechst33258 were comparable, indicating that the labeling for 191Pt via DTPA or Cys and the labeling for 111In via DTPA worked equally well. 191Pt showed a DNA-binding/cellular uptake ratio of more than 1 order of magnitude greater than that of 111In. [191Pt]Pt-Hoechst33258 labeled via Cys showed a higher cellular uptake than that labeled via DTPA, resulting in a very high DNA-binding fraction of [191Pt]Pt-Cys-Hoechst33258 and extensive DNA damage. Our labeling methods of radio-Pt, especially via Cys, promote the development of radio-Pt-based agents for use in Auger electron therapy targeting DNA.
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Cisteína , Ácido Pentético , Cisteína/química , ADN , Electrones , Ácido Pentético/químicaRESUMEN
A field study was conducted on the reuse of wastewater from Mardan city to evaluate its risk of contaminating soil and wheat grains at different NPK levels. Three irrigation sources i.e. waste water (WW), canal water (CW) and alternate waste + canal water (WW+CW) were applied to wheat (cv Atta Habib 2010) grown at 0, 50, 75 and 100% NPK levels of 120:90:60 kg N:P2O5:K2O ha-1 at Palatoo Research Farm, Amir Muhammad Khan Campus, Mardan during 2015.The results showed higher grain and biomass yields in WW irrigated plots as compared to CW at NPK levels up to 50% of recommending dose revealing supplementing nutrient requirements in deficient conditions. However, irrigation of WW at higher NPK levels especially at or beyond 75% of recommended dose tended to reduce the crop yield that could be associated with heavy metals toxicity and nutritional imbalances. The use of WW substantially increased AB-DTPA extractable Zn, Mn, Pb, Ni and Cd indicating a potential threat to soil contamination. Similarly, WW irrigated wheat had higher concentrations of these heavy metals as compared to CW which limits its use for production purposes without any remediation measures. The alternate use of CW and WW as revealed by its comparative lower contamination in soil and wheat than sole WW could be one of the possible solutions and may increase the time required for threshold soil contamination.
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Riego Agrícola , Álcalis/química , Monitoreo del Ambiente , Metales Pesados/análisis , Semillas/química , Suelo/química , Triticum/química , Aguas Residuales , Biomasa , Conductividad Eléctrica , Geografía , Concentración de Iones de Hidrógeno , Nitrógeno/análisis , Pakistán , Ácido Pentético/química , Fósforo/análisis , Potasio/análisisRESUMEN
The coexistence of field-induced blockage of the magnetization and significant magnetocaloric effects in the low-temperature region occurs in a mononuclear holmium(III) diethylenetriamine-N,N,N',Nâ³,Nâ³-pentaacetate complex, whose gadolinium(III) analogue is a commercial MRI contrast agent. Both properties make it a suitable candidate for cryogenic magnetic refrigeration, thus enlarging the variety of applications of this simple class of multifunctional molecular nanomagnets.
Asunto(s)
Complejos de Coordinación/química , Holmio/química , Imanes/química , Ácido Pentético/química , Refrigeración/métodos , Frío , Fenómenos MagnéticosRESUMEN
The diagnosis of gastrointestinal (GI) tract diseases is frequently performed in the clinic, so it is crucial to develop high-performance contrast agents for real-time and non-invasive imaging examination of the GI tract. Herein, we show a novel method to synthesize a neodymium (Nd) chelate, Nd-diethylenetriaminepentaacetic acid (Nd-DTPA), on a large scale without byproducts for spectral computed tomography (CT) and second near-infrared window imaging of the GI tract in vivo. The Nd-DTPA was simply generated by heating the mixture of Nd2O3 and DTPA in water at 85 °C for 2 h. This dual-modal imaging agent has the advantages of a simple and green synthesis route, no need of purification process, high yield (86.24%), large-scale production capability (>10 g in lab synthesis), good chemical stability and excellent water solubility (≈2 g mL-1). Moreover, the Nd-DTPA emitted strong near-infrared fluorescence at 1308 nm, and exhibited superior X-ray attenuation ability compared to clinical iohexol. The proposed Nd-DTPA can integrate the complementary merits of dual-modal imaging to realize spatial-temporal and highly sensitive imaging of the GI tract in vivo, and accurate diagnosis of the location of intestinal obstruction and monitor its recovery after surgery. The developed highly efficient method for the gram-scale synthesis of Nd-DTPA and the proposed spectral CT and second near-infrared window dual-modal imaging strategy provide a promising route for accurate visualization of the GI tract in vivo.
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Quelantes/química , Quelantes/síntesis química , Tracto Gastrointestinal/diagnóstico por imagen , Neodimio/química , Ácido Pentético/química , Tomografía Computarizada por Rayos X/métodos , Animales , Técnicas de Química Sintética , Femenino , Ratones , Solubilidad , Agua/químicaRESUMEN
The epithelial cell plays a key role in the transfer of radionuclides from lungs to blood following pulmonary exposure. The present study was designed to evaluate the transfer across human lung epithelial cells of various actinides (plutonium, americium and uranium), the influence of the physicochemical properties of plutonium compounds and of the chelating agent diethylene triamine pentaacetic acid (DTPA). To address this question, Calu-3 cells grown in a bicameral culture system were used. The integrity of the epithelial barrier was evaluated by measuring transepithelial electrical resistance (TEER) and the passage of a fluorescent marker, lucifer yellow. Activity measurement in basal compartment following periodic collection of culture medium was made from 2 h to seven days. To facilitate data handling and analysis, the statistical tool STATBIODIS was used. The results indicate differences in transfer for the different elements, and according to Pu physicochemical properties. Though to various extents, the chelating agent DTPA always increased the transfer of Pu and Am across the epithelial cells, without altering the integrity of the epithelial barrier. This in vitro cell culture model, by mimicking translocation of actinides from lungs to blood, can represent a valuable tool to further understand the underlying mechanisms and properties controlling this process.
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Elementos de Series Actinoides/farmacología , Quelantes/farmacología , Células Epiteliales/efectos de los fármacos , Ácido Pentético/farmacología , Elementos de Series Actinoides/química , Elementos de Series Actinoides/toxicidad , Transporte Biológico/efectos de los fármacos , Línea Celular Tumoral , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Quelantes/química , Quelantes/toxicidad , Células Epiteliales/metabolismo , Humanos , L-Lactato Deshidrogenasa/metabolismo , Pulmón/citología , Ácido Pentético/química , Ácido Pentético/toxicidadRESUMEN
Sorbus commixta is a valuable hardwood plant with a high economical value for its medicinal and ornamental qualities. The aim of this work was to investigate the effects of the iron (Fe) source and medium pH on the growth and development of S. commixta in vitro. The Fe sources used, including non-chelated iron sulfate (FeSO4), iron ethylenediaminetetraacetic acid (Fe-EDTA), and iron diethylenetriaminepentaacetic acid (Fe-DTPA), were supplemented to the Multipurpose medium with a final Fe concentration of 2.78 mg·L-1. The medium without any supplementary Fe was used as the control. The pH of the agar-solidified medium was adjusted to either 4.70, 5.70, or 6.70. The experiment was conducted in a culture room for six weeks with 25 °C day and night temperatures, and a 16-h photoperiod with a light intensity of 50 mmol·m-2·s-1 photosynthetic photon flux density (PPFD). Both the Fe source and pH affected the growth and development of the micropropagated plants in vitro. The leaves were greener in the pH 4.70 and 5.70 treatments. The tissue Fe content decreased with the increase of the medium pH. The leaf chlorophyll content was similar between plants treated with FeSO4 and those with Fe-EDTA. The numbers of the shoots and roots of plantlets treated with FeSO4 were 2.5 and 2 times greater than those of the control, respectively. The fresh and dry weights of the shoot and the root were the greatest for plants treated with Fe-EDTA combined with pH 5.70. The calcium, magnesium, and manganese contents in the plantlets increased in the pH 5.70 treatments regardless of the Fe source. Supplementary Fe decreased the activity of ferric chelate reductase. Overall, although the plantlets absorbed more Fe at pH 4.70, Fe-EDTA combined with pH 5.70 was found to be the best for the growth and development of S. commixta in vitro.
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Medios de Cultivo/farmacología , Compuestos Férricos/química , Compuestos Ferrosos/química , Ácido Pentético/análogos & derivados , Sorbus/crecimiento & desarrollo , Antioxidantes/química , Clorofila/química , Ácido Edético/química , FMN Reductasa/química , Concentración de Iones de Hidrógeno , Hierro , Ácido Pentético/química , Fotosíntesis , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismo , Estomas de Plantas/metabolismo , Sorbus/metabolismo , Factores de TiempoRESUMEN
A short (Fab)trastuzumab-derived peptide specific for HER2 receptor was identified. Its affinity for the model system HER2-DIVMP was found in a nanomolar range. The structural determinants responsible for the interaction between this ligand (A9) and HER2-DIVMP were investigated by both computational and NMR analysis. Next, the possibility of using A9 as HER2- specific probe for the nuclear medicine imaging was evaluated by conjugating A9 with the DTPA chelator and radiolabeling it with 111In. The developed probe retained a nanomolar affinity to HER2-overexpressing cancer cells, however, some unspecific binding also occurred. The peptide internalization into cells by receptor-mediated endocytosis was also studied. Future perspectives are aimed at using A9 as a probe for molecular imaging diagnostics as well as active targeting of anticancer drugs. Lead structure optimization is needed to minimize the percentage of A9 unspecific binding and to increase the binding affinity to the receptor.
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Péptidos/química , Receptor ErbB-2/metabolismo , Animales , Sitios de Unión , Humanos , Inmunoconjugados/química , Inmunoconjugados/metabolismo , Marcaje Isotópico , Ligandos , Imagen por Resonancia Magnética , Simulación de Dinámica Molecular , Neoplasias/diagnóstico por imagen , Neoplasias/metabolismo , Neoplasias/patología , Ácido Pentético/química , Péptidos/metabolismo , Unión Proteica , Receptor ErbB-2/agonistas , Receptor ErbB-2/antagonistas & inhibidores , Distribución Tisular , Trastuzumab/químicaRESUMEN
Site-specific labeling of proteins with a paramagnetic tag is an efficient way to provide atomic-resolution information about the dynamics, interactions, and structures of the proteins and protein-ligand complexes. The paramagnetic effects manifested in NMR spectroscopy generally contain paramagnetic relaxation enhancement, pseudocontact shifts (PCSs), and residual dipolar coupling (RDC), and these effects correlate closely with the flexibility of protein-tag conjugates. The rigidity of the paramagnetic tag is greatly important in decoding the structural details of macromolecular complexes, because paramagnetic averaging reduces the PCSs and RDCs. Here we show that the dynamic exchange of the metal chelating moiety is a key factor in determining the rigidity of the paramagnetic tag in the protein conjugates. Decreasing the conformational exchange rates in the metal chelating moiety greatly minimizes the paramagnetic averaging and thus increases PCSs and RDCs. This effect has been demonstrated in an open-chain tag, Py-l-Cys-DTPA, which generates large PCSs and RDCs that are comparable to those of the reported cyclic DOTA-like tags. The proposed route offers a unique way to design suitable paramagnetic tags for applications in biological systems.
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Quelantes/química , Complejos de Coordinación/química , Indicadores y Reactivos/química , Elementos de la Serie de los Lantanoides/química , Ubiquitina/química , Sitios de Unión , Compuestos Heterocíclicos con 1 Anillo/química , Cinética , Ligandos , Resonancia Magnética Nuclear Biomolecular , Ácido Pentético/química , Unión Proteica , Conformación ProteicaRESUMEN
One of the key components of radiopharmaceuticals for targeting imaging and therapy is a stable bifunctional chelating system to attach radionuclides to selective delivery systems. After-effects of radioactive decay can cause the release of a radioactive isotope from its chelation agent. Perturbed angular correlation (PAC) of γ-rays has become a unique technique to study the behavior of complexes formed between a chelating agent and radionuclide in vivo (in real time) over a relevant range of concentrations (10-12 M). In the present work, four radionuclides, 111In, 111mCd, and 152, 154Eu, were investigated with diethylenetriaminepentaacetic acid (DTPA) at different pH values to determine the stability constants of the complexes as well as the effects of post-decay processes, which play a major role in determining the suitability of these complexes for application as radiopharmaceuticals (e.g., in vivo generators). The study provides a convenient parameter for the characterization of radionuclide-chelator systems using the PAC method. PAC is proven to be a suitable tool to study novel chelators and radiopharmaceutical precursors attached to radiometals.
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Radioquímica/métodos , Radiofármacos/química , Rayos gamma , Concentración de Iones de Hidrógeno , Ácido Pentético/química , Radioisótopos/químicaRESUMEN
BACKGROUNDS: Due to the unexpected side effects of the iodinated contrast agents, novel contrast agents for X-ray computed tomography (CT) imaging are urgently needed. Nanoparticles made by heavy metal elements are often employed, such as gold and bismuth. These nanoparticles have the advantages of long in vivo circulation time and tumor targeted ability. However, due to the long residence time in vivo, these nanoparticles may bring unexpected toxicity and, the preparation methods of these nanoparticles are complicated and time-consuming. METHODS: In this investigation, a small molecular bismuth chelate using diethylenetriaminepentaacetic acid (DPTA) as the chelating agent was proposed to be an ideal CT contrast agent. RESULTS: The preparation method is easy and cost-effective. Moreover, the bismuth agent show better CT imaging for kidney than iohexol in the aspect of improved CT values. Up to 500 µM, the bismuth agent show negligible toxicity to L02 cells and negligible hemolysis. And, the bismuth agent did not induce detectable morphology changes to the main organs of the mice after intravenously repeated administration at a high dose of 250 mg/kg. The pharmacokinetics of the bismuth agent follows the first-order elimination kinetics and, it has a short half-life time of 0.602 h. The rapid clearance from the body promised its excellent biocompatibility. CONCLUSIONS: This bismuth agent may serve as a potential candidate for developing novel contrast agent for CT imaging in clinical applications.
Asunto(s)
Bismuto , Medios de Contraste , Tomografía Computarizada por Rayos X/métodos , Animales , Bismuto/química , Bismuto/farmacocinética , Bismuto/toxicidad , Medios de Contraste/química , Medios de Contraste/farmacocinética , Medios de Contraste/toxicidad , Yohexol/química , Yohexol/farmacocinética , Riñón/diagnóstico por imagen , Riñón/metabolismo , Nanopartículas del Metal/química , Nanopartículas del Metal/toxicidad , Ratones , Ácido Pentético/química , Ácido Pentético/farmacocinética , Distribución Tisular , Imagen de Cuerpo EnteroRESUMEN
PURPOSE: Radioimmunotherapy uses tumor-specific antibodies to deliver therapeutic radionuclides, but hematological toxicity due to the long serum half-life of intact antibodies remains a challenge. We evaluated a smaller antibody fragment, the minibody, with faster kinetics and a potentially improved therapeutic index. PROCEDURES: The anti-prostate stem cell antigen (PSCA) minibody (A11 Mb) was radiolabeled with iodine-124 ([124I]I-A11 Mb) or conjugated with deferoxamine (DFO) and labeled with zirconium-89 ([89Zr]Zr-DFO-A11 Mb) for surrogate immunoPET to profile pharmacokinetics in a human prostate cancer xenograft model. Subsequently, minibodies labeled with two therapeutic beta emitters, directly iodinated [131I]I-A11 Mb (non-residualizing) and 177Lu chelated using DTPA ([177Lu]Lu-DTPA-A11 Mb) (residualizing), were compared for in vitro antigen-specific cytotoxicity. Full biodistribution studies (in 22Rv1-PSCA tumor bearing and hPSCA knock-in mice) were conducted for dosimetry calculations. Finally, the lead candidate [131I]I-A11 Mb was evaluated in a radioimmunotherapy experiment. Escalating single doses (3.7, 11, or 37 MBq) and saline control were administered to 22Rv1-PSCA tumor bearing mice and anti-tumor effects (tumor volume) and toxicity (body weight) were monitored. RESULTS: Minibodies radiolabeled with therapeutic beta emitters [131I]I-A11 Mb and [177Lu]Lu-DTPA-A11 Mb exhibited comparable tumor cell growth inhibition in vitro. In vivo surrogate immunoPET imaging using [89Zr]Zr-DFO-A11 Mb showed activity retention in liver and kidney up to 72 h, while [124I]I-A11 Mb cleared from liver, kidney, and blood by 48 h. Based on full biodistribution and dosimetry calculations, administering 37 MBq [131I]I-A11 Mb was predicted to deliver a favorable dose to the tumor (35 Gy), with a therapeutic index of 22 (tumor:bone marrow). For [177Lu]Lu-DTPA-A11 Mb, the kidneys would be dose-limiting, and the maximum tolerated activity (7.4 MBq) was not predicted to deliver an effective radiation dose to tumor. Radioimmunotherapy with a single dose of [131I]I-A11 Mb showed dose-dependent tumor inhibition with minimal off-target toxicity and improved median survival (19 and 24 days, P < 0.001) compared with untreated mice (12 days). CONCLUSIONS: These findings show the potential of the anti-PSCA minibody for targeted radioimmunotherapy with minimal toxicity, and the application of immunoPET and dosimetry for personalized treatment.
Asunto(s)
Antígenos de Neoplasias/metabolismo , Radioisótopos de Yodo/química , Lutecio/química , Proteínas de Neoplasias/metabolismo , Ácido Pentético/química , Neoplasias de la Próstata/diagnóstico por imagen , Neoplasias de la Próstata/terapia , Radioinmunoterapia , Radioisótopos/química , Animales , Anticuerpos Monoclonales/farmacocinética , Línea Celular Tumoral , Proliferación Celular , Relación Dosis-Respuesta en la Radiación , Proteínas Ligadas a GPI/metabolismo , Masculino , Ratones , Ácido Pentético/farmacocinética , Neoplasias de la Próstata/inmunología , Radiometría , Análisis de Supervivencia , Distribución TisularRESUMEN
Development of adsorptive stripping voltammetry (AdSV) combined with in situ prepared bismuth film electrode (in situ BiFE) on glassy carbon disk surface using diethylenetriamine pentaacetic acid (DTPA) as a complexing agent and NO3 - as a catalyst to determine the trace amount of chromium (VI) is demonstrated. According to this method, in the preconcentration step at E dep = -800 mV, the bismuth film is coated on the surface of glassy carbon electrodes simultaneously with the adsorption of complexes Cr(III)-DTPA. In addition to the influencing factors, the stripping voltammetry performance factors such as deposition potential, deposition time, equilibration time, cleaning potential, cleaning time, and technical parameters of differential pulse and square wave voltammetries have been investigated, and the influence of Cr(III), Co(II), Ni(II), Ca(II), Fe(III), SO4 2-, Cl-, and Triton X has also been investigated. This method gained good repeatability with RSD <4% (n = 9) for the differential pulse adsorptive stripping voltammetry (DP-AdSV) and RSD < 3% (n = 7) for the square wave adsorptive stripping voltammetry (SqW-AdSV), and low limit of detection: LOD = 12.10-9 M ≈ 0.6 ppb (at a deposition potential (E dep) of -800 mV and the deposition time (t dep) of 50 s) and LOD = 2.10-9 M ≈ 0.1 ppb (at E dep = -800 mV and t dep = 160 s) for the DP-AdSV and SqW-AdSV, respectively. This method has been successfully applied to analyze chromium in natural water.