Development and evaluation of a 99mTc(III) ‘4 + 1’ complex derived from estradiol for imaging breast cancer

Estrogen receptors are overexpressed in about 70% of breast cancer and identification of their presence is important to select the appropriate treatment and evaluate the response.
With this objective, an estradiol derivative (L) 5-((1-carboxy-2-(4-((13S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-17-yl)-1H-1,2,3-triazol-1-yl)ethyl)amino)-N-methylidyne-5-oxopentan-1-aminium, was used to develop a ‘4+1’ complex of Tc(III) for estrogen receptor imaging.
The synthesis of L involved the coupling of the amino group of 3-azido-L-alanine with an activated isonitrile, to then perform a "click chemistry" reaction with the ethinyl group of ethinylestradiol.
Labelling was carried out in two stages, preparation of the precursor [99mTc]Tc-EDTA, using mannitol, EDTA and SnCl2 as reducing agent and simultaneous substitution (30 min at 75°C) with L (20 mg) and the tetradentate coligand 2-[Bis(2-mercaptoethyl)amino]ethanethiol (NS3) (2 mg). The HPLC analysis showed a major peak (tr=13 min). The radiochemical purity of the HPLC purified complex was greater than 95%.
The lipophilicity expressed as logP (partition coefficient between octanol and phosphate buffer 0.1M, pH = 7.4) was 0.48±0.06. The plasma protein binding was (46 ± 6) % at 60 minutes. The complex was stable in the labelling milieu and in human serum for at least 4 hours. Cell uptake in MCF7 cells are in progress.
A potential radiopharmaceutical derived from estradiol was obtained with high radiochemical purity. The complex presents adequate stability and physicochemical properties. In vitro and in vivo studies including nude mice bearing xenografted breast tumors will be used to validate the clinical potentiality.