ABSTRACT
Diazo transfer reactions on Behera's amine and its next-generation analogue formed G0 and G1 azide dendrons bearing three and nine tert-butyl-protected esters, respectively. The utility of the new dendrons was demonstrated by copper-catalyzed azide-alkyne cycloaddition, with 1,3,5-triethynylbenzene, forming two novel dendrimers in a convergent manner. Acid-mediated dendrimer deprotection was successful, and the resulting carboxy-terminated dendrimers were analyzed by NMR and DOSY experiments.
Subject(s)
Amines/chemistry , Azides/chemistry , Dendrimers/chemistry , Alkynes/chemistry , Catalysis , Copper/chemistry , Magnetic Resonance Spectroscopy , Molecular Structure , Polymers/chemistry , Solvents/chemistryABSTRACT
Important requirements for exogenous dyes or contrast agents in magnetic resonance imaging (MRI) include an effective concentration of paramagnetic or superparamagnetic ions at the target to be imaged. We report the concise synthesis and characterization of several new enantiopure bifunctional derivatives of (α(1)R,α(4)R,α(7)R,α(10)R)-α(1),α(4),α(7),α(10)-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTMA) (and their 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) analogues as controls) that can be covalently attached to a contrast agent delivery system using either click or peptide coupling chemistry. Gd complexes of these derivatives can be attached to delivery systems while maintaining optimal water residence time for increased molecular imaging sensitivity. Long chain biotin (LC-biotin) derivatives of the Eu(III) and Gd(III) chelates associated with avidin are used to demonstrate higher efficiencies. Variable-temperature relaxometry, (17)O NMR, and nuclear magnetic resonance dispersion (NMRD) spectroscopy used on the complexes and biotin-avidin adducts measure the influence of water residence time and rotational correlation time on constrained and unconstrained systems. The Gd(III)-DOTMA derivative has a shorter water residence time than the Gd(III)-DOTA derivative. Compared to the constrained Gd(III)-DOTA derivatives, the rotationally constrained Gd(III)-DOTMA derivative has â¼40% higher relaxivity at 37 °C, which could increase its sensitivity as an MRI agent as well as reduce the dose of the targeting agent.
Subject(s)
Chelating Agents/chemistry , Contrast Media/chemistry , Magnetic Resonance Imaging/methods , Biotin/chemistry , Gadolinium/chemistry , Models, Molecular , Quaternary Ammonium Compounds/chemistryABSTRACT
We evaluate novel magnetic resonance imaging (MRI) and positron emission tomography (PET) quantitative imaging biomarkers and associated multimodality, serial-time-point analysis methodologies, with the ultimate aim of providing clinically feasible, predictive measures for early assessment of response to cancer therapy. A focus of this work is method development and an investigation of the relationship between the information content of the two modalities. Imaging studies were conducted on subjects who were enrolled in glioblastoma multiforme (GBM) therapeutic clinical trials. Data were acquired, analyzed and displayed using methods that could be adapted for clinical use. Subjects underwent dynamic [(18)F]fluorothymidine (F-18 FLT) PET, sodium ((23)Na) MRI and 3-T structural MRI scans at baseline (before initiation of therapy), at an early time point after beginning therapy and at a late follow-up time point after therapy. Sodium MRI and F-18 FLT PET images were registered to the structural MRI. F-18 FLT PET tracer distribution volumes and sodium MRI concentrations were calculated on a voxel-wise basis to address the heterogeneity of tumor physiology. Changes in, and differences between, these quantities as a function of scan timing were tracked. While both modalities independently show a change in tissue status as a function of scan time point, results illustrate that the two modalities may provide complementary information regarding tumor progression and response. Additionally, tumor status changes were found to vary in different regions of tumor. The degree to which these methods are useful for GBM therapy response assessment and particularly for differentiating true progression from pseudoprogression requires additional patient data and correlation of these imaging biomarker changes with clinical outcome.
Subject(s)
Biomarkers/metabolism , Brain Neoplasms/diagnosis , Brain Neoplasms/therapy , Fluorine Radioisotopes/pharmacology , Glioblastoma/diagnosis , Glioblastoma/therapy , Magnetic Resonance Imaging/methods , Positron-Emission Tomography/methods , Sodium/pharmacology , Thymidine/pharmacology , Adult , Female , Humans , Image Processing, Computer-Assisted/methods , Male , Middle Aged , Sodium Isotopes/pharmacology , Thymidine Kinase/metabolism , Tissue DistributionABSTRACT
OBJECTIVES: We aimed to develop a directly detected magnetic resonance imaging (MRI) contrast agent for use with high fields based on a nanoscale fluorinated dendrimer-based platform for F MRI and overcome some of the problems with F MRI. MATERIALS AND METHODS: The dendrimers were prepared in a convergent manner by making the appropriate dendron, followed by coupling to a central core. The dendrons were prepared by attaching 3 equivalents of the fluorinated amino acid to the 3 carboxylic acids of the repeat branch unit followed by deprotection of the amine branch point, and either coupling to another repeat branch unit (increasing the generation G) or used directly allowing the precise growth of the dendrimer. The size of the dendrimers was determined by diffusion nuclear magnetic resonance (NMR) spectroscopy. The toxicity of the dendrimers was measured using the MTT assay. Fluorine longitudinal relaxation time measurements were performed on a Bruker ACP-500 NMR using a saturation recovery experiment at 470.59 MHz frequency. Healthy 150 g Sprague-Dawley female rats were imaged using a dendrimer solution. RESULTS: The size of the dendrimers is generally less than 3 nm, 2 orders of magnitude smaller than the size of the perfluorocarbon nanoparticles (about 200 nm). The longitudinal relaxation time, T1, decreases with increasing dendrimer generation. A significant improvement in relaxation rate and signal-to-noise ratio can be achieved by either the chemical modification of the dendrimer with a gadolinium-chelate or by the physical addition of exogenous contrast agent. Although the dendrimers with fluorine in the surface layer are toxic, this toxicity is easily reduced by burying the fluorine further into the dendrimer interior. (19)F MR images of the rat using the dendrimer solution were rapidly obtained at 7 Tesla, the strong contrast in the heart generated by the dendrimer can be seen. CONCLUSIONS: A novel fluorinated dendrimer-based nanotechnology platform in (19)F MRI and a new bifunctional DOTA chelate were prepared and characterized. We introduce 2 methods for reducing the (19)F longitudinal relaxation time: (a) Increasing the generation; (b) covalent and noncovalent introduction of Gd(III)-chelates. A new bifunctional Gd(III)-chelate is presented. The investigations of imaging on rats suggest potential importance of the dendrimers in (19)F MRI application.
Subject(s)
Contrast Media , Dendrimers/chemistry , Fluorocarbons/chemistry , Halogenation , Magnetic Resonance Imaging/methods , Nanotechnology/methods , Animals , Female , Gadolinium/chemistry , Magnetic Resonance Imaging/instrumentation , Magnetic Resonance Spectroscopy , Nanotechnology/instrumentation , Rats , Rats, Sprague-Dawley , Time FactorsABSTRACT
The title compound, C(26)H(45)N(3)O(8), is a bicyclic molecule; the seven-membered diazepane ring has a twisted-chair conformation and the six-membered morpholine ring has a boat conformation.
Subject(s)
Acetates/chemistry , Azepines/chemistry , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Morpholines/chemistry , Crystallography, X-Ray , Models, Molecular , Molecular Conformation , Molecular Structure , StereoisomerismABSTRACT
Epidermal growth factor receptor (EGFR) vIII is a mutated EGFR that is frequently overexpressed in glioblastomas and implicated in response to receptor tyrosine kinase inhibitors. In this study, we investigate the effect of ZD6474 (ZACTIMA, vandetanib), a dual inhibitor for vascular endothelial growth factor receptor 2 and EGFR on growth and angiogenesis of gliomas expressing EGFRvIII. We used two glioma xenograft models, U87MG cells overexpressing EGFRvIII and short-term cultured primary glioma GBM8 cells with EGFRvIII. ZD6474 inhibited tumor growth and angiogenesis and induced cell apoptosis in various brain gliomas. Moreover, significant inhibition of EGFRvIII-expressing U87MG and GBM8 gliomas was observed compared with their controls. Magnetic resonance imaging analysis using the apparent diffusion coefficient and three-dimensional T2*weighed measurements validated ZD6474 inhibition on tumor growth and angiogenesis in EGFRvIII-expressing GBM8 gliomas. Mechanistically, ZD6474 shows better inhibition of cell growth and survival of U87MG/EGFRvIII, GBM6, and GBM8 cells that express EGFRvIII than U87MG or GBM14 cells that have nondetectable EGFRvIII through attenuation of activated phosphorylation of signal transducer and activator of transcription 3, Akt, and Bcl-X(L) expression. Albeit in lesser extent, ZD6474 also displays suppressions of U87MG/EGFR and GBM12 cells that overexpress wild-type EGFR. Additionally, ZD6474 inhibits activation of extracellular signal-regulated kinase 1/2 in both types of cells, and expression of a constitutively active phosphoinositide 3-kinases partially rescued ZD6474 inhibition in U87MG/EGFRvIII cells. Taken together, these data show that ZD6474 significantly inhibited growth and angiogenesis of gliomas expressing EGFRvIII by specifically blocking EGFRvIII-activated signaling mediators, suggesting a potential application of ZD6474 in treatments for glioblastomas that overexpress EGFRvIII. Mol Cancer Ther; 9(4); 929-41. (c)2010 AACR.
Subject(s)
Brain/enzymology , ErbB Receptors/metabolism , Glioma/enzymology , Glioma/pathology , Mutant Proteins/metabolism , Piperidines/pharmacology , Quinazolines/pharmacology , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Animals , Brain/blood supply , Brain/drug effects , Brain/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Enzyme Activation/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Glioma/drug therapy , Humans , Magnetic Resonance Imaging , Mice , Neovascularization, Pathologic/drug therapy , Phosphorylation/drug effects , Piperidines/therapeutic use , Proto-Oncogene Proteins c-akt/metabolism , Quinazolines/therapeutic use , STAT3 Transcription Factor/metabolism , Time Factors , Treatment Outcome , Xenograft Model Antitumor Assays , bcl-X Protein/metabolismABSTRACT
Dendritic cells (DCs) are the most effective antigen-presenting cells (APCs) and are used in a variety of immunotherapeutic approaches. Adoptive cellular immunotherapy (ACI) of cancer using DCs has attracted much interest due to their capacity to promote immunity in prophylactic and therapeutic protocols. As one approach, DCs are injected into patients or tumor-bearing animals, to trigger specific antitumor immunity. In that framework, several approaches to DC delivery have been reported, including direct intratumoral injection; this has yielded positive but variable results. The underlying reasons for this have not been fully determined, but major hypotheses include technical difficulties in delivering cells into tumors and tumor-mediated immunosuppression. Image-guided ACI offers the potential to establish that DCs are efficiently delivered to the tumor site, which might eliminate some of the variability. Therefore, we developed highly sensitive methods for monitoring the injection or trafficking of DCs into tumors using a clinically approved formulation of a gadolinium-based magnetic resonance imaging (MRI) contrast agent, Gd(III)-HP-DO3A (ProHance). We determined the labeling efficiency of DCs with this formulation; that labeling DCs with this agent did not inhibit expression of surface markers important for antigen presentation and activation of naive T cells; that their capacity to interact with natural killer (NK) cells was not reduced; and that their migration was not diminished. Further, we determined that ProHance-labeled DCs can be effectively imaged in vivo in established central nervous system tumors.
Subject(s)
Brain Neoplasms/pathology , Brain Neoplasms/surgery , Dendritic Cells/pathology , Dendritic Cells/transplantation , Heterocyclic Compounds , Immunotherapy, Adoptive/methods , Magnetic Resonance Imaging/methods , Organometallic Compounds , Animals , Brain Neoplasms/immunology , Cells, Cultured , Contrast Media , Dendritic Cells/immunology , Gadolinium , Image Enhancement/methods , Magnetics , Male , Rats , Rats, Inbred F344 , Reproducibility of Results , Sensitivity and Specificity , Surgery, Computer-Assisted/methodsABSTRACT
PURPOSE: The purpose of this study was to compare histologically determined cellularity and extracellular space to dynamic contrast-enhanced magnetic resonance imaging (DCE MRI)-based maps of a two-compartment model's parameters describing tumor contrast agent extravasation, specifically tumor extravascular extracellular space (EES) volume fraction (ve), tumor plasma volume fraction (vp) and volume-normalized contrast agent transfer rate between tumor plasma and interstitium (KTRANS/VT). MATERIALS AND METHODS: Obtained ve, vp and KTRANS/VT maps were estimated from gadolinium diethylenetriamine penta-acetic acid DCE T1-weighted gradient-echo images at resolutions of 469, 938 and 2500 microm. These parameter maps were compared at each resolution to histologically determined tumor type, and the high-resolution 469-microm maps were compared with automated cell counting using Otsu's method and a color-thresholding method for estimated intracellular (Vintracellular) and extracellular (Vextracellular) space fractions. RESULTS: The top five KTRANS/VT values obtained from each tumor at 469 and 938 microm resolutions are significantly different from those obtained at 2500 microm (P<.0001) and from one another (P=.0014). Using these top five KTRANS/VT values and the corresponding tumor EES volume fractions ve, we can statistically differentiate invasive ductal carcinomas from noninvasive papillary carcinomas for the 469- and 938-microm resolutions (P=.0017 and P=.0047, respectively), but not for the 2500-microm resolution (P=.9008). The color-thresholding method demonstrated that ve measured by DCE MRI is statistically similar to histologically determined EES. The Vextracellular obtained from the color-thresholding method was statistically similar to the ve measured with DCE MRI for the top 10 KTRANS/VT values (P>.05). DCE MRI-based KTRANS/VT estimates are not statistically correlated with histologically determined cellularity. CONCLUSION: DCE MRI estimates of tumor physiology are a limited representation of tumor histological features. Extracellular spaces measured by both DCE MRI and microscopic analysis are statistically similar. Tumor typing by DCE MRI is spatial resolution dependent, as lower resolutions average out contributions to voxel-based estimates of KTRANS/VT. Thus, an appropriate resolution window is essential for DCE MRI tumor diagnosis. Within this resolution window, the top KTRANS/VT values with corresponding ve are diagnostic for the tumor types analyzed in this study.
Subject(s)
Magnetic Resonance Imaging/methods , Mammary Neoplasms, Animal/pathology , Animals , Contrast Media/pharmacokinetics , Female , Gadolinium DTPA/pharmacokinetics , Image Processing, Computer-Assisted , Rats , Rats, Sprague-DawleyABSTRACT
We tested the hypothesis that partial volume effects due to poor in-plane resolution and/or low temporal resolution used in clinical dynamic contrast-enhanced magnetic resonance imaging results in erroneous diagnostic information based on inaccurate estimates of tumor contrast agent extravasation and tested whether reduced encoding techniques can correct for dynamic data volume averaging. Image spatial resolution was reduced from 469 x 469 microm2 to those reported below by selecting a subset of k-space data. We then compared the top five K(trans)/V(T) "hot spots" obtained from the original data set, 469 x 469-microm in-plane spatial resolution and an 18-s temporal resolution processed by fast Fourier transform (FFT), with values obtained from data sets having in-plane spatial resolutions of 938 x 938, 1875 x 1875 and 2500 x 2500 microm2 and a temporal resolution of 18 s, or data sets with temporal resolutions of 36, 54 and 72 and a spatial resolution of 469 x 469 microm2, and found them to statistically differ from the parent data sets. We then tested four different post processing methods for improving the spatial resolution without sacrificing temporal resolution: zero-filled FFT, keyhole, reduced-encoding imaging by generalized-series reconstruction (RIGR) and two-reference RIGR (TRIGR). The top five values of K(trans)/V(T) obtained from data sets, the in-plane spatial resolutions of which were improved to 469 x 469 microm2 by zero-filling FFT, Keyhole and RIGR, statistically differed from those obtained from the original 469 x 469 microm2 FFT parent image data set. Only the 938 x 938 and 1875 x 1875 microm2 data sets reconstructed to 469 x 469 microm2 with TRIGR reconstruction method yielded values of the top five K(trans)/V(T) hot spots statistically the same as the original parent data set, 469 x 469 microm2 in-plane spatial and 18-s temporal-resolution FFT. That is, partial volume effects from data sets of different in-plane spatial resolution resulted in statistically different values of the top five K(trans)/V(T) hot spots relative to a high spatial and temporal resolution data set, and TRIGR reconstruction of these low resolution data sets to high resolution images provided statistically similar values with a savings in temporal resolution of 2 to 4 times.
Subject(s)
Magnetic Resonance Imaging/methods , Mammary Neoplasms, Experimental/pathology , Mammography/methods , Animals , Contrast Media , Female , Humans , Image Processing, Computer-Assisted , Magnetic Resonance Imaging/statistics & numerical data , Mammography/statistics & numerical data , Rats , Rats, Sprague-DawleyABSTRACT
RATIONALE AND OBJECTIVES: Dendrimer-based magnetic resonance imaging (MRI) contrast agents offer many advantages including high levels of amplification. The objective of this research was to test the adequacy and viability of a new family of dendrimers for use as MRI contrast agents in vitro and in vivo. METHODS: Dendrimers based on 1,4-diaminobutane core polypropyleneimine (PPI) generation 2 and ammonia core polyamidoamine dendrimers had the free surface amines conjugated to a diethylenetriaminepentaacetic acid derivative followed by complex formation with gadolinium. Relaxivity measurements were made on an IBM Field Cycling Relaxometer. Biodistribution and pharmacokinetic studies were examined with the radiotracer 153Gd in rats and a counting window of 95 to 105 keV. MRI images were conducted at 4.7 T. RESULTS: The relaxivity of the PPI agent exceeded that of the corresponding generation polyamidoamine (PAMAM) agent. Uptake occurred in the liver, spleen, and kidney. Pharmacokinetic studies showed a biexponential decay with excretion half-lives of 3 hours and 33.6 days respectively. The agent increased the contrast enhancement, 1 hour after injection, of T1-weighted images by 52%. CONCLUSIONS: This PPI agent resulted in significant contrast signal enhancement. This family of agent may also provide a valuable contrast agent backbone.
Subject(s)
Contrast Media/pharmacokinetics , Magnetic Resonance Imaging/methods , Pentetic Acid/analogs & derivatives , Spermidine/analogs & derivatives , Animals , Female , Gadolinium DTPA , Kidney/metabolism , Liver/metabolism , Macromolecular Substances , Models, Animal , Pentetic Acid/pharmacokinetics , Rats , Rats, Sprague-Dawley , Spleen/metabolismSubject(s)
Carrier Proteins/metabolism , Contrast Media , Heterocyclic Compounds/pharmacokinetics , Magnetic Resonance Imaging , Organometallic Compounds/pharmacokinetics , Ovarian Neoplasms/pathology , Receptors, Cell Surface , Animals , Carrier Proteins/genetics , Female , Folate Receptors, GPI-Anchored , Gadolinium/pharmacokinetics , Mice , Mice, Nude , Radioligand Assay , Transfection , Transplantation, Heterologous , Tumor Cells, CulturedABSTRACT
RATIONALE AND OBJECTIVES: Dynamic contrast enhanced (DCE) MR mammography (MRM) uses tumor capillary density differences for prognosis. The heterogeneous response of permeability-surface area products (PS = Kp<-->t) was examined in mammary tumors, as a function of contrast agent size, to determine what effect ROI size might have on PS and prognosis. METHODS: DCE FLASH signal intensities were converted to gadolinium concentrations by a standard curve, which was fitted by a two-compartment model for the tumor's extravascular extracellular space (EES) volume fraction (ve), and the tumor volume normalized transfer rate between plasma and EES (Kp<-->t/VT). RESULTS: For Gd-DTPA ve = 9% to 13% Kp<-->t/VT = 0.01 to 0.06 minutes-1, and the macromolecular agent, PAMAM-TU-DTPA G = 4 ve = 0.8% to 1% Kp<-->t/VT = 0.008 to 0.04 minutes(-1). Significant differences in Kp<-->t/VT for local regions were found for both agents relative to the whole tumor and the macromolecular agent had greater dynamic range. CONCLUSIONS: Smaller ROI values or pixels should yield more accurate assessment of neovascularization.
Subject(s)
Gadolinium DTPA , Magnetic Resonance Imaging/methods , Mammary Neoplasms, Experimental/blood supply , Mammary Neoplasms, Experimental/pathology , Polyamines , Animals , Capillary Permeability , Contrast Media/pharmacokinetics , Dendrimers , Ethylnitrosourea , Gadolinium DTPA/pharmacokinetics , Macromolecular Substances , Mathematics , Microcirculation , Polyamines/pharmacokinetics , Rats , Rats, Sprague-DawleyABSTRACT
RATIONALE AND OBJECTIVES: An important characteristic of targeted contrast agents is how they are tolerated in a biologic environment and their localization in the surrounding tissues in addition to target tissue. We evaluate the biodistribution of a gadolinium Gd 153-folate-dendrimer in high affinity folate-receptor (hFR) positive and negative ovarian tumor xenografts. METHODS: The 153Gd-folate-dendrimer chelate was prepared by exchanging 153Gd with nonradioactive gadolinium for 1 week, followed by extensive filtration. Athymic mice with hFR-positive (n = 3) and negative tumors (n = 3) were injected intravenously and counted using a whole-body counting system with a 80 to 150 keV counting window. RESULTS: The hFR-positive tumors accumulate 3.6% +/- 2.8% injected dose/g, whereas only background counts were found in hFR-negative tumors. The folate-dendrimer's tumor-to-blood ratio of 12.6, in hFR-positive tumors, was approximately 5.7 to 17.0 fold better than those obtained with monoclonal antibodies targeted to the folate receptor. CONCLUSIONS: Biodistribution studies confirm previous MRI findings and show that the accumulation of the folate-dendrimer requires the expression of the hFR.