Diego Alberti

MRI Visualization of Melanoma Cells by Targeting Overexpressed Sialic Acid with a GdIII‐dota‐en‐pba Imaging Reporter

These changes in glycosylation are mainly caused byan increased expression of sialyltransferases, which glycosy-late exposed glycans at their terminal positions with anionicmonosaccharide sialic acid residues (Sia). Tumor cells over-expressing Sia appear protected against the immune defensesystem, and as a result, malignancy is increased.

MRI-guided neutron capture therapy by use of a dual gadolinium/boron agent targeted at tumour cells through upregulated low-density lipoprotein transporters.

The upregulation of low-density lipoprotein (LDL) transporters in tumour cells has been exploited to deliver a sufficient amount of gadolinium/boron/ligand (Gd/B/L) probes for neutron capture therapy, a binary chemio-radiotherapy for cancer treatment. The Gd/B/L probe consists of a carborane unit (ten B atoms) bearing an aliphatic chain on one side (to bind LDL particles), and a Gd(III)/1,4,7,10-tetraazacyclododecane monoamide complex on the other (for detection by magnetic resonance imaging (MRI)). Up to 190 Gd/B/L probes were loaded per LDL particle. The uptake from tumour cells was initially assessed on cell cultures of human hepatoma (HepG2), murine melanoma (B16), and human glioblastoma (U87). The MRI assessment of the amount of Gd/B/L taken up by tumour cells was validated by inductively coupled plasma-mass-spectrometric measurements of the Gd and B content. Measurements were undertaken in vivo on mice bearing tumours in which B16 tumour cells were inoculated at the base of the neck. From the acquisition of magnetic resonance images, it was established that after 4-6 hours from the administration of the Gd/B/L-LDL particles (0.1 and 1 mmol kg(-1) of Gd and (10)B, respectively) the amount of boron taken up in the tumour region is above the threshold required for successful NCT treatment. After neutron irradiation, tumour growth was followed for 20 days by MRI. The group of treated mice showed markedly lower tumour growth with respect to the control group.

15N Magnetic Resonance Hyperpolarization via the Reaction of Parahydrogen with 15N-Propargylcholine

(15)N-Propargylcholine has been synthesized and hydrogenated with para-H(2). Through the application of a field cycling procedure, parahydrogen spin order is transferred to the (15)N resonance. Among the different isomers formed upon hydrogenation of (15)N-propargylcholine, only the nontransposed derivative contributes to the observed N-15 enhanced emission signal. The parahydrogen-induced polarization factor is about 3000. The precise identification of the isomer responsible for the observed (15)N enhancement has been attained through a retro-INEPT ((15)N-(1)H) experiment. T(1) of the hyperpolarized (15)N resonance has been estimated to be ca. 150 s, i.e., similar to that reported for the parent propargylcholine (144 s). Experimental results are accompanied by theoretical calculations that stress the role of scalar coupling constants (J(HN) and J(HH)) and of the field dependence in the formation of the observed (15)N polarized signal. Insights into the good cellular uptake of the compound have been gained.

Design of PLGA based nanoparticles for imaging guided applications.

An amphiphilic Gd(III) complex has been efficiently loaded in polylactic-co-glycolic acid nanoparticles (PLGA-NPs) to yield a novel, high sensitive magnetic resonance imaging (MRI) contrast agent for imaging guided drug delivery applications. As the Gd(III) complex is soluble in organic solvents, the nanoparticles were prepared as oil/water emulsions. PLGA-NPs were stable, in buffer, for more than 1 week without any release of the incorporated agents. The millimolar relaxivity of the Gd(III) complex incorporated in the particles (140 nm diameter) was of 21.7 mM(-1) s(-1) at 21.5 MHz, a value that is about 5 times higher than that observed with the commercially available contrast agents used in clinic. The relaxometric efficiency of these particles resulted inversely proportional to the particle size measured by dynamic light scattering. The high stability and sensitivity of PLGA-NPs allowed their accumulation in vivo in murine melanoma xenograft as shown in the corresponding MR images. Once loaded with drug and contrast agents, PLGA nanoparticles can be proposed as efficient theranostic MRI agents.

A Carborane‐Derivative “Click” Reaction under Heterogeneous Conditions for the Synthesis of a Promising Lipophilic MRI/GdBNCT Agent

In this study, the Huisgen reaction has been used to functionalise a carborane cage with a lipophilic moiety and a 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) ligand to obtain a new Gd boron neutron-capture therapy (BNCT)/magnetic resonance imaging (MRI) agent. The introduction of the triazole units has been accomplished under both heterogeneous conditions, by the use of a Cu-supported ionic-liquid catalyst, and homogeneous conditions. The ability of the Gd complex of the synthesised ligand to form stable adducts with low-density lipoproteins (LDLs) has been evaluated and then MRI has been performed on tumour melanoma cells incubated in the presence of a Gd-complex/LDL imaging probe. It has been concluded that the high amount of intracellular boron necessary to perform BNCT can be reached even in the presence of a relatively low-boron-containing LDL concentration.

MRI of cells and mice at 1 and 7 Tesla with Gd-targeting agents: when the low field is better!

Tumor cells were targeted with Gd-loaded/LDL (low density lipoproteins) adducts consisting of ca 300 Gd(III) amphiphilic complexes incorporated in the lipophilic LDL particles. The long reorientational time of the Gd(III) complex in the supramolecular adduct yielded a relaxivity peak at ca 1 T, whereas its relaxivity at 7 T was 5 times less. The field-dependent relaxivity markedly affected the signal enhancement attainable at the two magnetic fields. As tumor cells showed up-regulation of LDL transporters, B16 melanoma cells were labeled with the Gd-loaded/LDL adduct. Each cell contained ca 2 × 10(9) Gd atoms. Upon dispersion of 5000 labeled cells in 1 μl of agar, signal intensity (SI) enhancements of about 30 and 7% were observed at 1 and 7 T, respectively. The results obtained on cellular systems were confirmed in vivo upon the administration of Gd-loaded/LDL particles to C57 mice bearing a transplanted melanoma (B16) tumor. From the herein reported results, one may conclude that, for slowly moving Gd complexes, it is possible to obtain in vivo sensitivity enhancements at 1 T several times higher than that attained at high fields.

Synthesis of a carborane-containing cholesterol derivative and evaluation as a potential dual agent for MRI/BNCT applications

In this study the synthesis and characterization of a new dual, imaging and therapeutic, agent is proposed with the aim of improving the efficacy of Boron Neutron Capture Therapy (BNCT) in cancer treatment. The agent (Gd-B-AC01) consists of a carborane unit (ten boron atoms) bearing a cholesterol unit on one side (to pursue the incorporation into the liposome bi-layer) and a Gd(iii)/1,4,7,10-tetraazacyclododecane monoamide complex on the other side (as a MRI reporter to attain the quantification of the B/Gd concentration). In order to endow the BNCT agent with specific delivery properties, the liposome embedded with the MRI/BNCT dual probes has been functionalized with a pegylated phospholipid containing a folic acid residue at the end of the PEG chain. The vector allows the binding of the liposome to folate receptors that are overexpressed in many tumor types, and in particular, in human ovarian cancer cells (IGROV-1). An in vitro test on IGROV-1 cells demonstrated that Gd-B-AC01 loaded liposomes are efficient carriers for the delivery of the MRI/BNCT probes to the tumor cells. Finally, the BNCT treatment of IGROV-1 cells showed that the number of surviving cells was markedly smaller when the cells were irradiated after internalization of the folate-targeted GdB10-AC01/liposomes.

Boronated Compounds for Imaging Guided BNCT Applications

Boron neutron capture therapy (BNCT) is based on the capture of thermal neutrons by boron 10 ((10)B) nuclei that have been selectively delivered to tumor cells. The amount of 10-30 μg of boron for g of tumor mass is needed to attain an acceptable therapeutic advantage. Despite that the potentialities of BNCT have been demonstrated in several preclinical studies, this technique has not yet been fully accepted in the armory of tools for tumor treatment. This is partly due to the differences in the uptake and distribution of (10)B among patients and also to the uncertainty found in the determination of tumor-to-blood (10)B concentration ratio. Attention is now being payed to use the main imaging techniques to determine the in vivo biodistribution of BNCT agents. Most of the work has been devoted to the most promising BNCT agents, namely BPA, BSH and carborane derivatives. This review surveys studies carried out over the last decade, and outlines the role that NMR, PET and SPECT imaging may have to improve the efficacy of BNCT.

A theranostic approach based on the use of a dual boron/Gd agent to improve the efficacy of Boron Neutron Capture Therapy in the lung cancer treatment

This study aims at developing an innovative theranostic approach for lung tumor and metastases treatment, based on Boron Neutron Capture Therapy (BNCT). It relies on to the use of low density lipoproteins (LDL) as carriers able to maximize the selective uptake of boron atoms in tumor cells and, at the same time, to quantify the in vivo boron distribution by magnetic resonance imaging (MRI). Tumor cells uptake was initially assessed by ICP-MS and MRI on four types of tumor (TUBO, B16-F10, MCF-7, A549) and one healthy (N-MUG) cell lines. Lung metastases were generated by intravenous injection of a Her2+ breast cancer cell line (i.e. TUBO) in BALB/c mice and transgenic EML4-ALK mice were used as primary tumor model. After neutron irradiation, tumor growth was followed for 30-40 days by MRI. Tumor masses of boron treated mice increased markedly slowly than the control group. From the clinical editor: In this article, the authors described an improvement to existing boron neutron capture therapy. The dual MRI/BNCT agent, carried by LDLs, was able to maximize the selective uptake of boron in tumor cells, and, at the same time, quantify boron distribution in tumor and in other tissues using MRI. Subsequent in vitro and in vivo experiments showed tumor cell killing after neutron irradiation.

Theranostic Nanoparticles Loaded with Imaging Probes and Rubrocurcumin for Combined Cancer Therapy by Folate Receptor Targeting

The combination of different therapeutic modalities is a promising option to combat the recurrence of tumors. In this study, polylactic and polyglycolic acid nanoparticles were used for the simultaneous delivery of a boron–curcumin complex (RbCur) and an amphiphilic gadolinium complex into tumor cells with the aim of performing boron and gadolinium neutron capture therapy (NCT) in conjunction with the additional antiproliferative effects of curcumin. Furthermore, the use of Gd complexes allows magnetic resonance imaging (MRI) assessment of the amount of B and Gd internalized by tumor cells. Poly(lactic‐co‐glycolic acid) (PLGA) nanoparticles were targeted to ovarian cancer (IGROV‐1) cells through folate receptors, by including in the formulation a PEGylated phospholipid functionalized with the folate moiety. NCT was performed on IGROV‐1 cells internalizing 6.4 and 78.6 μg g−1 of 10B and 157Gd, respectively. The synergic action of neutron treatment and curcumin cytotoxicity was shown to result in a significant therapeutic improvement.