Porphyrin-substituted H-NOX proteins as high-relaxivity MRI contrast agents.

Heme proteins are exquisitely tuned to carry out diverse biological functions while employing identical heme cofactors. Although heme protein properties are often altered through modification of the protein scaffold, protein function can be greatly expanded and diversified through replacement of the native heme with an unnatural porphyrin of interest. Thus, porphyrin substitution in proteins affords new opportunities to rationally tailor heme protein chemical properties for new biological applications. Here, a highly thermally stable Heme Nitric oxide/OXygen binding (H-NOX) protein is evaluated as a magnetic resonance imaging (MRI) contrast agent. T1 and T2 relaxivities measured for the H-NOX protein containing its native heme are compared to the protein substituted with unnatural manganese(II/III) and gadolinium(III) porphyrins. H-NOX proteins are found to provide unique porphyrin coordination environments and have enhanced relaxivities compared to commercial small-molecule agents. Porphyrin substitution is a promising strategy to encapsulate MRI-active metals in heme protein scaffolds for future imaging applications.

Analysis of Lanthanide Complex Dendrimer Conjugates for Bimodal NIR and MRI Imaging.

Advances in clinical diagnostic instrumentation have enabled some imaging modalities to be run concurrently. For diagnostic purposes, multimodal imaging can allow for rapid location and accurate identification of a patient's illness. The paramagnetic and near Infra-red (NIR) properties of Dy(III) and Yb(III) are interesting candidates for the development of bimodal NIR and magnetic resonance imaging (MRI) contrast agents. To enhance their intrinsic bimodal properties, these lanthanides were chelated using the hexadentate-all-oxygen-donor-ligand TREN-bis-(1-Me)-3,2-HOPO-TAM-NX (NX, where X = 1, 2 or 3) and subsequently conjugated to the esteramide dendrimer (EA), to improve bioavailability, solubility, and relaxivity. Of these new complexes synthesized and evaluated, DyN1-EA had the largest ionic T(1) relaxivity, 7.60 mM(-1) s(-1), while YbN3-EA had the largest ionic T(2) relaxivity with a NIR quantum yield of 0.17 % when evaluated in mouse serum. This is the first Yb(III) bimodal NIR/T(2) MRI contrast agent of its kind evaluated.

Stealth rare earth oxide nanodiscs for magnetic resonance imaging.

Configuring RE(2) O(3) nanocrystals with pseudo-2D morphologies confers substantive gains in relaxivities over equivalent spherical counterparts. The most promising arises from Gd(2) O(3) whose ionic transverse and longitudinal relaxivities were as high as r(1) = 12.7 mM(-1) s(-1) and r(2) = 17.2 mM(-1) s(-1) (dispersed in H(2) O, T = 37 °C & B(0) = 1.41 T), pointing to new opportunities to achieve high contrast MRI while concomitantly affording longer half-life potential in vivo.

Silica microparticles as a solid support for gadolinium phosphonate magnetic resonance imaging contrast agents.

Particle-based magnetic resonance imaging (MRI) contrast agents have been the focus of recent studies, primarily due to the possibility of preparing multimodal particles capable of simultaneously targeting, imaging, and treating specific biological tissues in vivo. In addition, particle-based MRI contrast agents often have greater sensitivity than commercially available, soluble agents due to decreased molecular tumbling rates following surface immobilization, leading to increased relaxivities. Mesoporous silica particles are particularly attractive substrates due to their large internal surface areas. In this study, we immobilized a unique phosphonate-containing ligand onto mesoporous silica particles with a range of pore diameters, pore volumes, and surface areas, and Gd(III) ions were then chelated to the particles. Per-Gd(III) ionic relaxivities ranged from ∼2 to 10 mM(-1) s(-1) (37 °C, 60 MHz), compared to 3.0-3.5 mM(-1) s(-1) for commercial agents. The large surface areas allowed many Gd(III) ions to be chelated, leading to per-particle relaxivities of 3.3 × 10(7) mM(-1) s(-1), which is the largest value measured for a biologically suitable particle.

Improving T1 and T2 magnetic resonance imaging contrast agents through the conjugation of an esteramide dendrimer to high-water-coordination Gd(III) hydroxypyridinone complexes.

Commercial gadolinium magnetic resonance imaging (MRI) contrast agents are limited by low relaxivity (r1) and coordination to only a single water molecule (q = 1). Consequently, gram quantities of these agents must be injected to obtain sufficient diagnostic contrast. In this study, MRI contrast agents for T(1) and T2 relaxivity were synthesized using hydroxypyridinone and terephthalamide chelators with mesityl and 1,4,7-triazacyclononane capping moieties. When covalently conjugated to a highly biocompatible esteramide dendrimer, T2 relaxation rates up to 52 mm(-1) s(-1) and T1 relaxation rates up to 31 mm(-1) s(-1) per gadolinium were observed under clinically relevant conditions. These values are believed to be brought about by using a dendritic macromolecule to decrease the molecular tumbling time of the small molecule complexes. These agents also show high aqueous solubility and low toxicity in vitro. In this study we report six new compounds: three discrete complexes and three dendrimer conjugates.

Conjugation to Biocompatible Dendrimers Increases Lanthanide T2 Relaxivity of Hydroxypyridinone (HOPO) Complexes for Magnetic Resonance Imaging (MRI).

Magnetic resonance imaging (MRI) contrast agents represent a worldwide billion-dollar market annually. While T1 relaxivity enhancement contrast agents receive greater attention and a significantly larger market share, the commercial potential for T2 relaxivity enhancing contrast agents remains a viable diagnostic option due to their increased relaxivity at high field strengths. Improving the contrast and biocompatibility of T2 MRI probes may enable new diagnostic prospects for MRI. Paramagnetic lanthanides have the potential to decrease T1 and T2 proton relaxation times, but are not commercially used in MRI diagnostics as T2 agents. In this article, oxygen donor chelates (hydroxypyridinone, HOPO, and terephthalamide, TAM) of various lanthanides are demonstrated as biocompatible macromolecular dendrimer conjugates for the development of T2 MRI probes. These conjugates have relaxivities up to 374 mm-1s-1 per dendrimer, high bioavailability, and low in vitro toxicity.

Conjugation effects of various linkers on Gd(III) MRI contrast agents with dendrimers: optimizing the hydroxypyridinonate (HOPO) ligands with nontoxic, degradable esteramide (EA) dendrimers for high relaxivity.

One essential requirement for more sensitive gadolinium-based MRI contrast agents is to slow the molecular tumbling of the gadolinium(III) ion, which increases the gadolinium's relaxivity (i.e., its ability to speed up the NMR relaxation of nearby water molecules). One route to this is through conjugation to high-molecular-weight polymers such as dendrimers. In this work, amine-functionalized TREN-bis(1,2-HOPO)-TAM-ethylamine and TREN-bis(1-Me-3,2-HOPO)-TAM-ethylamine ligands have been synthesized and attached to biocompatible 40 kDa esteramide (EA)- and poly-l-lysine (PLL)-based dendrimers capable of binding up to eight gadolinium complexes. These conjugates have T(1) relaxivities of up to 38.14 ± 0.02 mM(-1) s(-1) per gadolinium at 37 °C, corresponding to relaxivities of up to 228 mM(-1) s(-1) per dendrimer molecule. This relaxivity expressed on a "per Gd" basis is several times that of the small-molecule complexes and an order of magnitude higher than that of current commercial agents. Because of their high performance and low toxicity, these macromolecules may constitute an attractive complement to currently available gadolinium(III)-based contrast agents.