Target-specific Contrast Agents Research Articles

A Polypeptide-Based, Membrane-Penetrating, Target-Specific Contrast Agent for Magnetic Resonance Molecular Imaging.

Molecular imaging based on magnetic resonance imaging (MRI) requires a contrast agent with high relaxivity and specificity. Much effort has been devoted to this goal over the past decades. In this work, we continue this endeavor by synthesizing an MRI contrast agent that can penetrate the cellular membrane and bind with specific proteins. It was characterized with one- and two-dimensional NMR spectroscopy, NMR micro-imaging, and mass spectroscopy. The target specificity has been further confirmed by both molecular dynamics simulation and micro-imaging on a living biological system. It is one of the largest of peptide-based bioactivated MRI contrast agents, and its relaxivity enhancement factor is among the highest of MRI contrast agents hitherto published. We envision interesting applications and extension of this smart MRI contrast agent with bio-specificity and high contrast for molecular imaging.

Strategies for Target-Specific Contrast Agents for Magnetic Resonance Imaging

This review describes recent research efforts focused on increasing the specificity of contrast agents for proton magnetic resonance imaging (MRI). Contrast agents play an indispensable role in MRI by enhancing the inherent contrast of images; however, the non-specific nature of current clinical contrast agents limits their usefulness. This limitation can be addressed by conjugating contrast agents or contrast-agent-loaded carriers-including polymers, nanoparticles, dendrimers, and liposomes-to molecules that bind to biological sites of interest. An alternative approach to conjugation is synthetically mimicking biological structures with metal complexes that are also contrast agents. In this review, we describe the advantages and limitations of these two targeting strategies with respect to translation from in vitro to in vivo imaging while focusing on advances from the last ten years.

Accelerating three‐dimensional molecular cardiovascular MR imaging using compressed sensing

To accelerate the acquisition of three-dimensional (3D) high-resolution cardiovascular molecular MRI by using Compressed Sensing (CS) reconstruction. Molecular MRI is an emerging technique for the early assessment of cardiovascular disease. This technique provides excellent soft tissue differentiation at a molecular and cellular level using target-specific contrast agents (CAs). However, long scan times are required for 3D molecular MRI. Parallel imaging can be used to speed-up these acquisitions, but hardware considerations limit the maximum acceleration factor. This limitation is important in small-animal studies, where single-coils are commonly used. Here we exploit the sparse nature of molecular MR images, which are characterized by localized and high-contrast biological target-enhancement, to accelerate data acquisition. CS was applied to detect: (a) venous thromboembolism and (b) coronary injury and aortic vessel wall in single- and multiple-coils acquisitions, respectively. Retrospective undersampling showed good overall image quality with accelerations up to four for thrombus and aortic images, and up to three for coronary artery images. For higher acceleration factors, features with high CA uptake were still well recovered while low affinity targets were less preserved with increased CS undersampling artifacts. Prospective undersampling was performed in an aortic image with acceleration of two, showing good contrast and well-defined tissue boundaries in the contrast-enhanced regions. We demonstrate the successful application of CS to preclinical molecular MR with target specific gadolinium-based CAs using retrospective (accelerations up to four) and prospective (acceleration of two) undersampling.

Dual-labeling strategies for nuclear and fluorescence molecular imaging: a review and analysis.

Molecular imaging is used for the detection of biochemical processes through the development of target-specific contrast agents. Separately, modalities such as nuclear and near-infrared fluorescence (NIRF) imaging have been shown to non-invasively monitor disease. More recently, merging of these modalities has shown promise owing to their comparable detection sensitivity and benefited from the development of dual-labeled imaging agents. Dual-labeled agents hold promise for whole-body and intraoperative imaging and could bridge the gap between surgical planning and image-guided resection with a single, molecularly targeted agent. In this review, we summarized the literature for dual-labeled antibodies and peptides that have been developed and have highlighted key considerations for incorporating NIRF dyes into nuclear labeling strategies. We also summarized our findings on several commercially available NIRF dyes and offer perspectives for developing a toolkit to select the optimal NIRF dye and radiometal combination for multimodality imaging.

Early Detection of Breast Cancer: Synthesis and Characterization of Novel Target Specific NIR-Fluorescent Estrogen Conjugate for Molecular Optical Imaging

Estrogen induced proliferation of existing mutant cells is widely understood to be the major risk determining factor in the development of breast cancer. Hence determination of the Estrogen Receptor[ER] status is of paramount importance. We have carried out the synthesis and characterization of a novel NIR fluorescent dye conjugate aimed at measuring ER+ve status in-vivo. The conjugate was synthesized by ester formation between 17-β estradiol and a cyanine dye namely: bis-1, 1-(4-sulfobutyl) indotricarbocyanine-5-carboxylic acid, sodium salt. The replacement of the sodium ion in the ester by a larger glucosammonium ion was found to enhance the hydrophilicity and reduce the toxic effect on cell lines. The excitation and emission peaks for the dye were recorded as 750 and 788 nm respectively; ideal for non-invasive optical imaging owing to minimal tissue attenuation and auto-fluorescence at these wavelengths. The dye (NIRDC1) has a significant drop in plasma-protein binding therefore leading to marked improvement in pharmacokinetic profile such as dye evacuation in comparison to ICG. In addition the dye showed enhanced fluorescence quantum yield, molar extinction coefficient and linearity in fluorescence relative to ICG. This dye can be potentially used as a target specific exogenous contrast agent in molecular optical imaging for early detection of breast cancer.

Nanoparticles containing octreotide peptides and gadolinium complexes for MRI applications

New mixed nanoparticles were obtained by self-aggregation of two amphiplic monomers. The first monomer (C18)(2) L5-Oct contains two C18 hydrophobic moieties bound to the N-terminus of the cyclic peptide octreotide, and spaced from the bioactive peptide by five units of dioxoethylene linkers. The second monomer, (C18)(2) DTPAGlu, (C18)(2) DTPA or (C18)(2) DOTA, and the corresponding Gd(III) complexes, contains two C18 hydrophobic moieties bound through a lysine residue to different polyamino-polycarboxy ligands: DTPAGlu, DTPA or DOTA. Mixed aggregates have been obtained and structurally characterized by small angle neutron scattering (SANS) techniques and for their relaxometric behavior. According to a decrease of negative charges in the surfactant head-group, a total or a partial micelle-to-vesicle transition is observed by passing from (C18)(2) DTPAGlu to (C18)(2) DOTA. The thicknesses of the bilayers are substantially constant, around 50 Å, in the analyzed systems. Moreover, the mixed aggregates, in which a small amount of amphiphilic octreotide monomer (C18)(2) L5-Oct (10% mol/mol) was inserted, do not differ significantly from the respective self-assembled systems. Fluorescence emission of tryptophan residue at 340 nm indicates low mobility of water molecules at the peptide surface. The proton relaxivity of mixed aggregates based on (C18)(2) DTPAGlu(Gd), (C18)(2) DTPA(Gd) and (C18)(2) DOTA(Gd) resulted to be 17.6, 15.2 and 10.0 mM(-1) s(-1) (at 20 MHz and 298K), respectively. The decrease in the relaxivity values can be ascribed to the increase in τ(M) (81, 205 and 750 ns). The presence of amphiphilic octreotide monomer exposed on mixed aggregate surface gives the entire nanoparticles a potential binding selectivity toward somatostatin sstr2 receptor subtype, and these systems could act as MRI target-specific contrast agent.

Vascular Endothelial Growth Factor Receptor 2-Specific Microbubbles for Molecular Ultrasound Detection of Prostate Cancer in a Rat Model

To investigate whether rat prostate cancer can be detected by means of molecular ultrasound (US) using target-specific microbubbles versus a nonspecific contrast agent. A total of 20 Copenhagen rats were randomly examined 75 to 104 days after orthotopic implantation of G-Dunning rat prostatic tumor cells was using a high-end US system and either 1.2 mL/kg of the nonspecific agent A or 0.1 mL/kg of the target-specific agent B, containing vascular endothelial growth factor receptor 2 binding peptide. Contrast inflow (areas under the curve analysis) was determined during the first 30s, and all microbubbles were destroyed in the scan plane by means of the flash technique 20 minutes after intravenous administration to investigate specific accumulation of individual bubbles in tumors. Differences between normal prostate tissue and tumor were analyzed using luminance images. Sonographically determined tumor localization and extent were compared with magnetic resonance imaging and histology. The median tumor size in the 20 rats determined on US (2.3 mm) and magnetic resonance imaging (2.4 mm) showed a very high correlation (r = 0.92, P = 0.01). Both agent A and agent B demonstrated higher vascularization of tumor periphery compared with normal prostate (P < 0.05) based on contrast inflow and areas under the curve analysis. Twenty minutes after administration, luminance for agent B in the tumor was significantly higher (P = 0.003) compared with nonspecific agent A (11.8-0.1). In consensus reading, the increase in signal intensity of the tumor compared with normal prostate tissue was significantly higher for agent B (P = 0.005), whereas no significant difference was found for agent A (P = 0.180). The target-specific contrast agent was superior to the unspecific US contrast agent both with regard to early inflow analysis and specific accumulation in prostate cancer after 20 minutes.

MR molecular imaging of HER‐2 in a murine tumor xenograft by SPIO labeling of anti‐HER‐2 affibody

In vivo molecular imaging is a rapidly growing research area both for basic and clinical science. Non-invasive imaging of in vivo conditions at the molecular level increases understanding of the biological characteristics of normal and diseased tissues without the need for invasive surgical procedures. Among the various imaging modalities, magnetic resonance imaging (MRI) has garnered interest as a molecular imaging modality due to its high spatial resolution. Here, we have demonstrated that the combined use of HER-2 targeting affibody, a small 7 kDa molecule that behaves similarly to antibodies, and superparamagnetic iron oxide (SPIO) can non-invasively image HER-2 expressing cells or tissues both in vitro and in vivo by MRI. This preliminary study demonstrates that affibody-SPIO is a feasible, target-specific contrast agent for in vivo MR molecular imaging.

Molecular Magnetic Resonance Imaging of Myocardial Perfusion With EP-3600, a Collagen-Specific Contrast Agent

Cardiac magnetic resonance (MR) perfusion imaging during the first pass after intravenous administration of extracellular contrast agents is hampered by the spatial and temporal resolution achievable and by the artifacts seen in ultrafast MR imaging. Furthermore, time-consuming quantitative data analysis is often added. The use of molecular MR imaging with a target-specific contrast agent with perfusion-dependent binding to myocardium may enable prolonged visualization of perfusion defects and thus may help to overcome limitations of currently used first-pass extracellular MR imaging. EP-3600 is a new gadolinium-containing molecular contrast agent that binds reversibly to myocardial collagen. A significant but nonocclusive coronary artery stenosis was modeled in 7 domestic swine with an undersized MR-compatible balloon positioned in the left anterior descending artery as verified by x-ray angiography. Two animals died before contrast injection as a result of arrhythmias. In 5 swine, high-spatial-resolution gradient echo imaging (approximately 1 x 1 mm(2) in-plane resolution) was performed before and 5, 20, 40, and 60 minutes after intravenous administration of 12.3 micromol/kg EP-3600. Contrast was administered during stress induced by an infusion of 250 mumol x kg(-1) x min(-1) adenosine. Yb-DTPA was administered simultaneously for comparison of myocardium-to-plasma ratios. Images were assessed subjectively by 2 investigators, and signal-to-noise and contrast-to-noise ratios over time were calculated. Normal myocardium showed a significant signal-to-noise ratio increase during the entire examination time. In all animals (n=5), the perfusion defect in the left anterior descending artery territory could be visualized with a high contrast-to-noise ratio for at least 20 minutes after contrast injection. A significantly higher myocardium-to-plasma ratio was found for EP-3600 compared with the control agent Yb-DTPA (0.85+/-0.26 versus 0.22+/-0.08, respectively; P<0.01). EP-3600 is a new molecular MR imaging contrast agent that binds to the myocardium and enables prolonged, high-contrast, high-spatial-resolution visualization of myocardial perfusion defects.

Target-specific contrast agents for magnetic resonance microscopy

High-resolution ex vivo magnetic resonance (MR) imaging can be used to delineate prominent architectonic features in the human brain, but increased contrast is required to visualize more subtle distinctions. To aid MR sensitivity to cell density and myelination, we have begun the development of target-specific paramagnetic contrast agents. This work details the first application of luxol fast blue (LFB), an optical stain for myelin, as a white matter-selective MR contrast agent for human ex vivo brain tissue. Formalin-fixed human visual cortex was imaged with an isotropic resolution between 80 and 150 μm at 4.7 and 14 T before and after en bloc staining with LFB. Longitudinal (R1) and transverse (R2) relaxation rates in LFB-stained tissue increased proportionally with myelination at both field strengths. Changes in R1 resulted in larger contrast-to-noise ratios (CNR), per unit time, on T1-weighted images between more myelinated cortical layers (IV–VI) and adjacent, superficial layers (I–III) at both field strengths. Specifically, CNR for LFB-treated samples increased by 229±13% at 4.7 T and 269±25% at 14 T when compared to controls. Also, additional cortical layers (IVca, IVd, and Va) were resolvable in 14T-MR images of LFB-treated samples but not in control samples. After imaging, samples were sliced in 40-micron sections, mounted, and photographed. Both the macroscopic and microscopic distributions of LFB were found to mimic those of traditional histological preparations. Our results suggest target-specific contrast agents will enable more detailed MR images with applications in imaging pathological ex vivo samples and constructing better MR atlases from ex vivo brains.

Nano Imaging Technologies: Polymer vesicles loaded with precipitated gadolinium nanoparticles: A novel target-specific contrast agent for magnetic resonance imaging

Nano Imaging Technologies: Polymer vesicles loaded with precipitated gadolinium nanoparticles: A novel target-specific contrast agent for magnetic resonance imaging

Moderne MR-Verfahren zur Beurteilung der Tumorresponse

Magnetic resonance imaging (MRI) is a mainstay in oncological imaging with respect to tumor detection, characterization and treatment monitoring. Besides quantifying metric changes of tumor tissue, a wide range of different other surrogate parameters of therapy response can be imaged and quantified by MRI. Early monitoring of treatment success is critical both for medical and economical reasons specifically with more expensive target-specific drugs entering the clinical arena. Dynamic contrast-enhanced (DCE) and steady state MRI can help to assess tumor perfusion and vessel permeability. The cellular state of tissue can be measured by diffusion-weighted imaging (DWI) and metabolic changes can be monitored by MR spectroscopy (MRS). New target-specific contrast agents potentially allow selective imaging of apoptotic events.This review aims to give a brief overview of new MR-based imaging approaches to assess tumor response to new target-specific therapy regimes, with special emphasis on anti-angiogenic and antivascular treatment effects.

Synthesis, binding affinity, and relaxivity of target-specific MRI contrast agents

Although magnetic resonance imaging (MRI) is one of the most important imaging modalities of the central nervous system (CNS), one of the main drawbacks of MRI is its limited specificity. This can potentially be partially alleviated by target-specific contrast agents. In the present paper we describe a simple high yield synthesis of two such gadolinium-based spiperone targeted MRI contrast agents, 1a and 1b. The R1 relaxivities of 1a and 1b were evaluated and found to be 5.94 and 8.31 mM−1 s−1, respectively at 9.4T, while their R2 relaxivities at the same magnetic field were found to be 18.05 and 22.60 mM−1 s−1, respectively. In addition and very importantly compound 1a, which is a gadolinium-based, spiperone-targeted MRI contrast agent, was found to preserve some of the spiperone affinity toward the dopamine D2 receptor. Compounds 1a and 1b thus represent potential agents for in vitro dopamine receptor imaging using MRI in experimental models.

Synthesis and characterization of iron oxide/polymer composite nanoparticles with pendent functional groups

We describe in this paper an approach to synthesize superparamagnetic iron oxide nanoparticles in the presence of polymerized lactic acid. The resulted particles consisted of clusters of iron oxide monocrystals, embedded inside the polymer chains. The composite particles synthesized in situ were highly dispersible in aqueous solution with good stability. X-ray diffraction and magnetometer data all confirmed the crystalline structure and super-paramagnetic property of the particles. They exhibited narrow size distribution with hydrodynamic diameters close to 80 nm. In addition, the particles were shown to have abundant surface carboxyl groups, which can be used to conjugate various biomolecules. Such a preparation would be especially useful for developing target specific MRI contrast agents or drug delivery vehicles.

Möglichkeiten Target-spezifischer molekularer Bildgebung mit Ultraschallkontrastmitteln

Non-invasive molecular imaging technologies provide researchers with the opportunity to study cellular and molecular processes. Among different imaging technologies, ultrasound based molecular imaging methods are also of interest, since the use of ultrasound contrast agents allows specific and sensitive depiction of molecular targets. Recent studies are encouraging and have demonstrated the feasibility of ultrasound based molecular imaging. This review summarizes current experiences and recent preclinical studies with target-specific ultrasound contrast agents.

Cardiovascular magnetic resonance imaging of coronary atherothrombosis.

Atherothrombosis, defined as atherosclerotic lesion disruption with superimposed thrombus formation, is the major cause of acute coronary syndromes and cardiovascular death. CMR imaging of coronary atherothrombosis is challenging because of the small caliber of the vessels combined with respiratory and cardiac motion. Freebreathing 3D CMR coronary vessel wall imaging has enabled in vivo quantification of coronary plaque burden and remodeling as a marker of subclinical CAD. Molecular imaging via novel target-specific contrast agents such as fibrin-binding agents to detect arterial thrombus shows great promise as the new frontier in noninvasive imaging. Advances in molecular imaging and CMR techniques offer the potential for direct imaging of coronary thrombosis and in-stent thrombosis by use of novel fibrin-binding molecular MR contrast agents. Although the current role of noninvasive CMR imaging of atherothrombosis remains investigational, these techniques should enhance our understanding of the natural history of acute coronary syndromes and thereby facilitate strategies to prevent acute coronary syndromes and cardiovascular death in vulnerable patients.

Design and Synthesis of a Bimodal Target-Specific Contrast Agent for Angiogenesis

[structure: see text] A bimodal target-specific contrast agent based on a cyclic peptide containing the target-specific NGR sequence, gadolinium(III) diethylenetriaminepentaacetic acid (Gd(III)DTPA), and Oregon Green 488 (OG488) suitable for both MR imaging and optical imaging of angiogenesis is developed. The synthetic strategy for this target-specific contrast agent exploits the use of highly efficient, chemoselective reactions, such as native chemical ligation, and gives a straightforward approach for double labeling of peptides in general.

Mixed Micelles Composed of Peptides and Gadolinium Complexes as Tumor-Specific Contrast Agents in MRI: A SANS Study

A detailed structural investigation on mixed micelle aggregates as target-specific contrast agents for magnetic resonance imaging technique has been carried out by way of small angle neutron scattering measurements. These mixed micelles are formed by two new amphiphilic molecules formed by a bioactive peptide and a claw moiety. The first molecule, C18H37CONH(AdOO)x-G-CCK8 (C18LxCCK8, x = 2, 5), contains an 18-carbon-atom alkylic chain bound to the C-terminal of the cholecystokinin octapeptide amide (CCK 26−33 or CCK8) and is able to bind to the overexpressed CCK receptor of some tumor cells. The second molecule, C18H37CONHLys(DTPAGlu)CONH25- (C18DTPAGlu) or its gadolinium complex [C18H37CONHLys(DTPAGlu)CONH2Gd]2-, C18DTPAGlu(Gd), contains the same 18-carbon-atom alkylic chain bound, through a lysine residue, to the DTPAGlu chelating agent. Small angle neutron scattering measurements have been performed on ternary systems at different total concentrations and at various ratios of the two molecules. The effect of the concentration on the aggregation number as well as on the shape of the micelle has been investigated. Furthermore in order to optimize the exposure of the peptide on the micelle surface, C18LxCCK8 having the spacer L of different length has been used. The pure binary systems of the synthesized molecules are also presented.

Physicochemical properties of mixed micellar aggregates containing CCK peptides and Gd complexes designed as tumor specific contrast agents in MRI.

New amphiphilic molecules containing a bioactive peptide or a claw moiety have been prepared in order to obtain mixed micelles as target-specific contrast agents in magnetic resonance imaging. The first molecule, C(18)H(37)CONH(AdOO)(2)-G-CCK8 (C18CCK8), contains a C18 hydrophobic moiety bound to the C-terminal cholecystokinin octapeptide amide (CCK 26-33 or CCK8). The second amphiphilic compound, C(18)H(37)CONHLys(DTPAGlu)CONH(2) (C18DTPAGlu) or its gadolinium complex, (C18DTPAGlu(Gd)), contains the same C18 hydrophobic moiety bound, through a lysine residue, to the DTPAGlu chelating agent. The mixed aggregates as well as the pure C18DTPAGlu aggregate, in the presence and absence of Gd, have been fully characterized by surface tension measurements, FT-PGSE-NMR, fluorescence quenching, and small-angle neutron scattering measurements. The structural characterization of the mixed aggregates C18DTPAGlu(Gd)-C18CCK8 indicates a spherical arrangement of the micelles with an external shell of approximately 21 A and an inner core of approximately 20 A. Both the DTPAGlu(Gd) complexes and the CCK8 peptides point toward the external surface. The measured values for relaxivity in saline medium at 20 MHz proton Larmor frequency and 25 degrees C are 18.7 mM(-)(1) s(-)(1). These values show a large enhancement in comparison with the isolated DTPAGlu(Gd) complex.

Molecular targeting of Alzheimer's amyloid plaques for contrast‐enhanced magnetic resonance imaging

Smart molecular probes for both diagnostic and therapeutic purposes are expected to provide quantum advances in clinical medicine and biomedical research. We describe such a probe that targets amyloid plaques of Alzheimer's disease and has been labelled with gadolinium to impart contrast detectable by magnetic resonance imaging. Although amyloid plaques are a primary pathologic feature of Alzheimer's disease and amyloid reduction is a major therapeutic objective in Alzheimer's disease, no method currently exists for directly imaging human amyloid plaques in vivo. However, with the aid of a target specific contrast agent, imaging of amyloid plaques is theoretically possible with magnetic resonance imaging. We describe experiments proving that three essential properties exist in the smart molecular probe, putrescine‐gadolinium‐AB peptide: (1) transport across the blood–brain barrier following intravenous injection conferred by the polyamine moiety (2) binding to plaques with high affinity by putrescine‐AB, and (3) magnetic resonance imaging contrast by gadolinium. The plaque‐to‐background tissue contrast‐to‐noise ratio was enhanced more than 9 fold in all regions of cortex and hippocampus following intravenous administration of putrescine‐gadolinium‐AB in Alzheimer's disease transgenic mice. The ability of this magnetic resonance contrast probe to image plaques in vivo may enable early diagnosis and also provide a direct measure of the efficacy of antiamyloid therapies currently being developed. With the appropriate target specific probe (e.g. antibody), this technique could be applied to many other central nervous system disorders and targets.