Citation
Mohd Ashhar, Zarif Naim
(2020)
Preparation and characterization of [68Ga]NODAGA-Pamidronic acid for pet bone cancer imaging.
Masters thesis, Universiti Putra Malaysia.
Abstract
Early detection of bone metastases is essential to prevent skeletal-related
events. Unlike biopsies, a non-invasive technique to diagnose bone metastases
is by utilizing radiopharmaceuticals and detected using a nuclear imaging
modality. Hence, this research deems to determine the role of gallium-68
radiolabeled bisphosphonates ([68Ga]NODAGA-Pamidronic acid) for PET bone
cancer imaging. This study aims to ascertain the preparation, characterization,
and radiolabeling of [68Ga]NODAGA-Pamidronic acid. Lastly, to determine its
potential application, the in vitro bone binding assay and in vivo bone-to-blood
ratio is examined. Firstly, NODAGA-Pamidronic acid (NODPAM) was prepared
via the NHS ester conjugation method and characterized using tandem mass
spectrometry (MS/MS). The RP-HPLC method was then developed to remove
the free NODAGA using 0.1% trifluoroacetic acid and water as the mobile phase
at a flow rate of 0.5 ml/ min. Based on the MS/MS analysis of NODPAM, the
precursor ion and product ion observed were according to the theoretical value
(theoretical [M-H]-m/z: 591.14, obtained [M-H]-m/z: 591.14, [M-H-H3PO3]-m/z:
509.17). The isotopic abundance M+1 (calculated m/z: 22.02, obtained m/z:
20.99±0.94) confirms the molecular formula C18H34N4O14P2. The HPLC method
developed shows a good separation between peaks with a resolution of 1.613.
The freeze-dried NODPAM produces a solid white powder. Next, the
radiolabeling of [68Ga]NODPAM was optimized by looking into three parameters;
pH, temperature, and amount NODPAM. Finally, the in vitro bone binding assay
and in vivo bone-to-blood ratio was determined using synthetic hydroxyapatite
and Sprague Dawley rats, respectively. From the results obtained, the %RCP of
radiolabeled [68Ga]NODPAM was above 90% within 15 minutes at pH 4-4.5 and
a temperature of above 60ºC. The in vitro hydroxyapatite (HA) bone binding
assay displayed a significant difference between the [68Ga]NODPAM
82.25%±1.72 and [99mTc]MDP of 53.21%±0.28 (p<0.05). The bone-to-blood ratio
of [68Ga]NODPAM 2-hour post-injection was significantly higher (P<0.05)
compared to 68Ga(III); 27.53 and 0.74, respectively. In conclusion,
[68Ga]NODPAM was prepared and characterized accordingly, and the in vitro
bone binding assay and in vivo bone-to-blood ratio were assessed. The
preliminary data suggests that there is a need for a complete pre-clinical study
of [68Ga]NODPAM before translating it into clinical research.
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