[Dy(EOB-DO3A)] is prepared according to the general
synthetic methods, and characterized by spectroscopic analysis.Therelaxivities
that measured at 9.4 T animal MRI are r1
= 1.01, r2 = 2.80 mM-1s-1.We
observe acceptablenegative-enhancement with liver T2-weighted image,
alsoconfirm about 30% liver accumulation within 1 h post-injection at
inductively coupled plasma (ICP) spectrometer data.
Materials and Methods
All reagents were purchased from commercial sources and used as received. Characterization of the synthesized compounds have been determined by analytical spectroscopic methods.(1) T1 measurements were carried out using an RARE-inversion recovery method with variable inversion times (TI) at1.5 T (64 MHz, GE healthcare, USA), 9.4T (400 MHz, bruker, Germany) both. For T2, the multi-slice multi-echo (MSME) sequence was adaptedwith 70 echoes at 1.5 T, 9.4 T. R1, R2 relaxation times were obtained from the nonlinear least-squares fit of the mean pixel values at variable TI and TE, respectively. The relaxivities (r1 and r2) were then calculated as a slope of linear fit of relaxation rate along with concentrations. The in vivo T2 weighted MR image was obtained at 9.4 T animal MRI equipment (bruker, BioSpec 94/20, Germany). In these studies, the mice (20 ~ 23 g) were anesthetized by 1.5% isoflurane in oxygen. MR images were acquired before and after intravascular injection of [Dy(EOB-DO3A)] (dose: 0.2mmolDy/kg). Also image parameters of axial images for fast spin-echo, RARE are as follows: Repetition time (TR) = 2500 msec; echo time (TE) = 18.33msec; 35 X 35 X 30 mm field of view (FOV); 128 X 128 X 30 matrix size; 1.0 mm slice thickness; RARE factor = 8; scan time of each image = 40 sec. Biodistribution of [Dy(EOB-DO3A)] was assessed with ICP-MS, ICR mice (6 weeks, 21~24 g) were used in this study.(2)Result and Discussion
Scheme 1 shows the synthesis of [Dy(EOB-DO3A)], and the high-resolution FAB mass was obtained with this complex. Relaxivities of this dysprosium complex are r1 = 0.16, r2 = 0.16 mM-1s-1at low field MRI. But at high filed (9.4 T) are r1 = 1.01, r2 = 2.80 mM-1s-1. This field-dependence relaxivity is due to increasing of curie relaxation portion that increases over external magnetic field especially in T2 relaxation.(3) Unfortunately, r2value is lower than iron based nano-agent in high field. That results from the ligand (EOB-DO3A) structure that has carboxyclic acid arm as a strong electron withdrawing group. We are ongoing structure-activity studies with several ligands to overcome the low T2 relaxation. To assess the liver targeting property, ICP-MS of this complex was obtained according to post-injection time courses. Like in Figure 1, liver accumulation of the complex are about 30% within 1 h post-injection time. After 24 h, the complex is almost excreted from mice body. In vivo MRI experiment, [Dy(EOB-DO3A)] shows acceptable liver enhancement immediately after tail vein injection. As shown in Figure 2, it is excreted via urinal and hepatointestinal courses both. Figure 3 shows the axial T2-weighted MR images of hepatocellular carcinoma (HCC) model nude mice which were obtained by tail vein injection with [Dy(EOB-DO3A)]. The signal differencesbetween normal hepatocytes and tumor cells are obvious in post-injection image. Therefore, [Dy(EOB-DO3A)] demonstrated the probability to detect liver tumor and differentiate the margin between normal and abnormal liver cells.
Conclusion
We have successfully synthesized [Dy(EOB-DO3A)] as a high field liver targeting MRI CA.The results show that it might be useful as T2 agent in ultra-high field MRI. In addition, it is also demonstrated that [Dy(EOB-DO3A)] is valuable to diagnose liver cancer with hepatobiliary property.(1) Ah Rum Baek, J. Med. Chem.2017, 60, 4861-4868
(2) Hee-Kyung Kim, J. Med. Chem.2013, 56, 8104-8111
(3) Todd C. Soesbe, Magn. Reson. Med.2011, 66, 1697-1703