Tesis
Fe8-Based MRI Contrast Agents Targeted to Breast and Ovarian Cancer Cells
Autor
Parga Rivera, Kenia A.
Raptis, Raphael G. (Consejero)
Institución
Resumen
Magnetic resonance imaging (MRI) contrast agents (CA’s) are of great
interest for cancer imaging as a complementary tool in cancer diagnosis.
Tumor-targeting MRI-CA conjugates based on an [Fe8O4(pz)12Cl4]
paramagnetic cluster were synthesized to generate a breast/ovarian cancer
tumor-targeting CA. Fe8 clusters were chosen for their stability, and because
biochemical mechanisms for the efficient elimination of iron exist. Conjugates
of Fe8 and hyaluronic acid (HA) work by targeting CD44, a proteoglycan which
is over expressed on most human breast and ovarian cancer cell lines as well
as on tumor specimens, and is a major receptor for HA. The ultimate goal of
this project was to study the properties of an ovarian cancer tumor-targeting
CA.
Several Fe8-phenol complexes were prepared and characterized,
including Fe8-Tyr. Taking a cue from the demonstrated affinity of iron for
phenolate ligands, the synthesis and characterization of new Fe(III)-Tyr-HA
conjugates was subsequently developed, with Tyramine-substituted
Hyaluronic Acid (Tyr-HA). The conjugates produced are soluble in water. Tyr-
HA5 and Tyr-HA16 refer to the tyramine functionalized 5 KDa and 16 KDa
HA, respectively.
The synthetic work of the conjugates encompasses three steps, (1)
coupling of 4-hydroxyphenethylamine (Tyr) to the targeting group -- hyaluronic
acid (HA) to form Tyr-HA (Tyr-HA5 and Tyr-HA16) conjugates -- (2) synthesis
of the Fe8 complex, and (3) coupling of the Fe8 complex, by the substitution of a chloride terminal ligand, to the Tyr-HA conjugate to form the targeted Fe8-
Tyr-HA5 and Fe8-Tyr-HA16 CA’s.
The characterization of the conjugates was carried out by EDS, TEM,
IR, and Mössbauer spectroscopies, and TGA. The data generated led to the
following general conjugate formula for Fe8-Tyr-HA5:
(Fecubane)4(Feouter-Tyr-HA)1.6(Feouter-O-Feouter)2.4
Experimental studies in aqueous solution, such as pH, Dynamic Light
Scattering and Zeta Potential, demonstrate the conjugate’s stable behavior in
aqueous solution, with an average hydrodynamic diameter of 145 ± 0.4 nm
and 407.4 ± 0.5 nm, and a zeta potential of -28 ± 2 mV and -30 ± 3 for Fe8-
Tyr-HA5 and Fe8-Tyr-HA16, respectively.
The relaxivities of the Fe8-Tyr-HA conjugates were determined by their
T1 and T2 values in water. For Fe8-Tyr-HA5, r1 = 0.11 mM-1s-1 and r2 = 26.20
mM-1s-1 at 7.2 T, while r1 = 0.11 mM-1s-1 and r2 = 52.18 mM-1s-1 at 11.9 T. For
Fe8-Tyr-HA16, r1 = 0.39 mM-1s-1 and r2 = 0.10 mM-1s-1 at 7.2 T, while r1 = 0.47
mM-1s-1 and r2 = 0.23 mM-1s-1 at 11.9 T. The relaxivity values were compared
to the relaxivities of similar complexes in the current literature.
Preliminary in vivo experiments conducted using nude mice have
shown that the lead Fe8-Tyr-HA5 conjugate is physiologically compatible and
non-toxic in the dose ranges administered. Clear visibility of CA following an
intratumoral (direct) injection was observed; clear negative contrast in
circulating blood was evident, with attenuation persisting for more than 10
minutes following injection. Moreover, with an intravenous administration
heterogeneous signal change within the tumor was evident by 15 minutes after injection and was increasingly conspicuous with time over this window of
observations.