Mozaffari Saeed, Erfani Mostafa, Beiki Davood, Johari Daha Fariba, Kobarfard Farzad, Balalaie Saeed, Fallahi Babak
Department of Radiopharmacy, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Nuclear Science Research School, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of Iran (AEOI), Tehran, Iran.
Iran J Pharm Res. 2015 Winter;14(1):97-110.
Neurokinin 1 receptors (NK1R) are overexpressed on several types of important human cancer cells. Substance P (SP) is the most specific endogenous ligand known for NK1Rs. Accordingly,a new SP analogue was synthesized and evaluated for detection of NK1R positive tumors.[6-hydrazinopyridine-3-carboxylic acid (HYNIC)-Tyr(8)-Met(O)(11)-SP] was synthesized and radiolabeled with (99m)Tc using ethylenediamine-N,N'-diacetic acid (EDDA)and Tricine as coligands. Common physicochemical properties of radioconjugate were studied and in-vitro cell line biological tests were accomplished to determine the receptor mediated characteristics. In-vivo biodistribution in normal and tumor bearingnude mice was also assessed. The cold peptide was prepared in high purity (>99%) and radiolabeled with (99m)Tc at high specific activities (84-112GBq/µmol) with an acceptable labeling yield (>95%). The radioconjugate was stable in-vitro in the presence of human serum and showed 44% protein binding to human serumalbumin. In-vitro cell line studies on U373MG cells showed an acceptable uptake up to 4.91 ± 0.22% with the ratio of 60.21 ± 1.19% for its specific fraction and increasing specific internalization during 4 h. Receptor binding assays on U373MG cells indicated a mean Kd of 2.46 ± 0.43 nM and Bmax of 128925 ± 8145 sites/cell. In-vivo investigations determined the specific tumor uptake in 3.36 percent of injected dose per gram (%ID/g) for U373MG cells and noticeable accumulations of activity in the intestines and lung. Predominant renal excretion pathway was demonstrated. Therefore, this new radiolabeled peptide could be a promising radiotracer for detection of NK1R positive primary or secondary tumors.
神经激肽1受体(NK1R)在几种重要的人类癌细胞上过度表达。P物质(SP)是已知的NK1R最特异性的内源性配体。因此,合成了一种新的SP类似物,并对其检测NK1R阳性肿瘤的能力进行了评估。[6-肼基吡啶-3-羧酸(HYNIC)-Tyr(8)-Met(O)(11)-SP]被合成,并使用乙二胺-N,N'-二乙酸(EDDA)和三(羟甲基)甲基甘氨酸作为共配体用(99m)Tc进行放射性标记。研究了放射性偶联物的常见物理化学性质,并完成了体外细胞系生物学试验以确定受体介导的特性。还评估了在正常和荷瘤裸鼠体内的生物分布。冷肽以高纯度(>99%)制备,并以高比活(84-112GBq/µmol)用(99m)Tc进行放射性标记,标记产率可接受(>95%)。放射性偶联物在人血清存在下在体外稳定,与人血清白蛋白的蛋白结合率为44%。对U373MG细胞的体外细胞系研究显示,摄取率可达4.91±0.22%,其特异性部分的比例为60.21±1.19%,并在4小时内特异性内化增加。对U373MG细胞的受体结合试验表明,平均解离常数(Kd)为2.46±0.43nM,最大结合容量(Bmax)为128925±8145位点/细胞。体内研究确定,U373MG细胞的肿瘤特异性摄取为每克注射剂量的3.36%(%ID/g),且在肠道和肺部有明显的活性积聚。证明了主要的肾脏排泄途径。因此,这种新的放射性标记肽可能是检测NK1R阳性原发性或继发性肿瘤的有前景的放射性示踪剂。