{"title":"68Ga/ 177lu标记leuproliide肽类似物检测乳腺癌的合成、放射性标记和临床前评价","authors":"Subhani M Okarvi, Ibrahim Al-Jammaz","doi":"10.1089/cbr.2021.0370","DOIUrl":null,"url":null,"abstract":"<p><p><b><i>Objectives:</i></b> The expansion of novel and potent tumor receptor binding peptides is a promising approach for the precise targeting of various cancer. Leuprolide is a 9-residue peptide analog of gonadotropin-releasing hormone and is extensively used in the treatment of sex hormone-dependent tumors, including prostate, breast, and ovarian cancer. This preclinical study was undertaken to prepare a new radiolabeled leuprolide peptide for the detection of breast carcinoma. <b><i>Methods:</i></b> A 1,4,7,10-tetraazacyclododecane-<i>N,N</i>'<i>,N</i>''<i>,N</i>'''-tetraacetic acid (DOTA)-coupled 9-amino acid leuprolide peptide was synthesized after typical 9-fluorenylmethyl-oxycarbonyl-based solid-phase peptide synthesis and radiolabeled with both <sup>68</sup>Ga and <sup>177</sup>Lu radionuclides for theranostic use. The systemic pharmacokinetics was done in healthy balb/c mice. The <i>in vitro</i> tumor cell binding affinity was determined on MCF7, T47D, and MDA-MB-231 breast cancer cell lines. <i>In vivo</i> tumor targeting and micro positron-emission tomography imaging was performed on nude mice with MCF7 breast tumor xenografts. <b><i>Results:</i></b> The leuprolide peptide was conveniently synthesized by solid-phase synthesis strategy and its identity and purity were validated by mass spectrometry and high-performance liquid chromatography. The peptide radiolabeled efficiently (˃94%) with both diagnostic (<sup>68</sup>Ga) and therapeutic (<sup>177</sup>Lu) radionuclides and displayed nanomolar binding potency to all three tested MCF7, T47D, and MDA-MB-231 cell lines. Fast and favorable pharmacokinetics was observed for <sup>68</sup>Ga/<sup>177</sup>Lu-leuprolide in healthy Balb/c mice. In nude mice, <sup>68</sup>Ga-leuprolide peptide exhibited rapid clearance from the blood circulation with low to moderate (up to 5% ID/g) uptake/retention by the major body organs. The accumulation in the estrogen receptor-positive MCF7 tumor was 2.24% ± 0.62% ID/g at 45 min p.i, with good tumor to blood and muscle uptake ratios. The radiolabeled peptide was excreted primarily through the renal pathway. <b><i>Conclusion:</i></b> The encouraging results of this initial study demonstrate that additional testing of this leuprolide peptide seems to be indicated because of its convincing potential to be a new agent for the management of breast carcinoma.</p>","PeriodicalId":518937,"journal":{"name":"Cancer biotherapy & radiopharmaceuticals","volume":" ","pages":"372-383"},"PeriodicalIF":0.0000,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":"{\"title\":\"Synthesis, Radiolabeling, and Preclinical Evaluation of <sup>68</sup>Ga/<sup>177</sup>Lu-Labeled Leuprolide Peptide Analog for the Detection of Breast Cancer.\",\"authors\":\"Subhani M Okarvi, Ibrahim Al-Jammaz\",\"doi\":\"10.1089/cbr.2021.0370\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b><i>Objectives:</i></b> The expansion of novel and potent tumor receptor binding peptides is a promising approach for the precise targeting of various cancer. Leuprolide is a 9-residue peptide analog of gonadotropin-releasing hormone and is extensively used in the treatment of sex hormone-dependent tumors, including prostate, breast, and ovarian cancer. This preclinical study was undertaken to prepare a new radiolabeled leuprolide peptide for the detection of breast carcinoma. <b><i>Methods:</i></b> A 1,4,7,10-tetraazacyclododecane-<i>N,N</i>'<i>,N</i>''<i>,N</i>'''-tetraacetic acid (DOTA)-coupled 9-amino acid leuprolide peptide was synthesized after typical 9-fluorenylmethyl-oxycarbonyl-based solid-phase peptide synthesis and radiolabeled with both <sup>68</sup>Ga and <sup>177</sup>Lu radionuclides for theranostic use. The systemic pharmacokinetics was done in healthy balb/c mice. The <i>in vitro</i> tumor cell binding affinity was determined on MCF7, T47D, and MDA-MB-231 breast cancer cell lines. <i>In vivo</i> tumor targeting and micro positron-emission tomography imaging was performed on nude mice with MCF7 breast tumor xenografts. <b><i>Results:</i></b> The leuprolide peptide was conveniently synthesized by solid-phase synthesis strategy and its identity and purity were validated by mass spectrometry and high-performance liquid chromatography. The peptide radiolabeled efficiently (˃94%) with both diagnostic (<sup>68</sup>Ga) and therapeutic (<sup>177</sup>Lu) radionuclides and displayed nanomolar binding potency to all three tested MCF7, T47D, and MDA-MB-231 cell lines. Fast and favorable pharmacokinetics was observed for <sup>68</sup>Ga/<sup>177</sup>Lu-leuprolide in healthy Balb/c mice. In nude mice, <sup>68</sup>Ga-leuprolide peptide exhibited rapid clearance from the blood circulation with low to moderate (up to 5% ID/g) uptake/retention by the major body organs. The accumulation in the estrogen receptor-positive MCF7 tumor was 2.24% ± 0.62% ID/g at 45 min p.i, with good tumor to blood and muscle uptake ratios. The radiolabeled peptide was excreted primarily through the renal pathway. <b><i>Conclusion:</i></b> The encouraging results of this initial study demonstrate that additional testing of this leuprolide peptide seems to be indicated because of its convincing potential to be a new agent for the management of breast carcinoma.</p>\",\"PeriodicalId\":518937,\"journal\":{\"name\":\"Cancer biotherapy & radiopharmaceuticals\",\"volume\":\" \",\"pages\":\"372-383\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cancer biotherapy & radiopharmaceuticals\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1089/cbr.2021.0370\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/3/21 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cancer biotherapy & radiopharmaceuticals","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1089/cbr.2021.0370","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/3/21 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2
摘要
目的:扩增新型有效的肿瘤受体结合肽是一种有前途的精确靶向各种癌症的方法。Leuprolide是一种促性腺激素释放激素的9-残基肽类似物,广泛用于治疗性激素依赖性肿瘤,包括前列腺癌、乳腺癌和卵巢癌。本临床前研究是为了制备一种新的放射性标记的leuprolide肽,用于检测乳腺癌。方法:采用典型的9-氟甲基氧羰基固相肽合成方法,合成1,4,7,10-四氮杂环十二烷-N,N',N',N'' -四乙酸偶联的9-氨基酸leuprolide肽,并用68Ga和177Lu放射性核素进行放射性标记,用于治疗。在健康balb/c小鼠体内进行了全身药代动力学研究。测定其与MCF7、T47D和MDA-MB-231乳腺癌细胞株的体外肿瘤细胞结合亲和力。对移植MCF7乳腺肿瘤的裸鼠进行体内肿瘤靶向和微正电子发射断层成像。结果:采用固相合成方法合成了该leuprolide肽,并通过质谱和高效液相色谱验证了其纯度和特性。该肽对诊断核素(68Ga)和治疗核素(177Lu)的放射性标记均有效(94%),并对三种测试的MCF7、T47D和MDA-MB-231细胞系显示出纳摩尔结合能力。68Ga/177Lu-leuprolide在Balb/c健康小鼠体内具有良好的药代动力学。在裸鼠中,68Ga-leuprolide肽从血液循环中迅速清除,主要器官的吸收/保留低至中等(高达5% ID/g)。雌激素受体阳性MCF7肿瘤中积累量为2.24%±0.62% ID/g (45 min p.i),肿瘤对血液和肌肉的摄取比良好。放射性标记肽主要通过肾脏途径排泄。结论:这项初步研究的令人鼓舞的结果表明,对这种leuprolide肽进行额外的测试似乎是有必要的,因为它有令人信服的潜力成为一种治疗乳腺癌的新药物。
Synthesis, Radiolabeling, and Preclinical Evaluation of 68Ga/177Lu-Labeled Leuprolide Peptide Analog for the Detection of Breast Cancer.
Objectives: The expansion of novel and potent tumor receptor binding peptides is a promising approach for the precise targeting of various cancer. Leuprolide is a 9-residue peptide analog of gonadotropin-releasing hormone and is extensively used in the treatment of sex hormone-dependent tumors, including prostate, breast, and ovarian cancer. This preclinical study was undertaken to prepare a new radiolabeled leuprolide peptide for the detection of breast carcinoma. Methods: A 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA)-coupled 9-amino acid leuprolide peptide was synthesized after typical 9-fluorenylmethyl-oxycarbonyl-based solid-phase peptide synthesis and radiolabeled with both 68Ga and 177Lu radionuclides for theranostic use. The systemic pharmacokinetics was done in healthy balb/c mice. The in vitro tumor cell binding affinity was determined on MCF7, T47D, and MDA-MB-231 breast cancer cell lines. In vivo tumor targeting and micro positron-emission tomography imaging was performed on nude mice with MCF7 breast tumor xenografts. Results: The leuprolide peptide was conveniently synthesized by solid-phase synthesis strategy and its identity and purity were validated by mass spectrometry and high-performance liquid chromatography. The peptide radiolabeled efficiently (˃94%) with both diagnostic (68Ga) and therapeutic (177Lu) radionuclides and displayed nanomolar binding potency to all three tested MCF7, T47D, and MDA-MB-231 cell lines. Fast and favorable pharmacokinetics was observed for 68Ga/177Lu-leuprolide in healthy Balb/c mice. In nude mice, 68Ga-leuprolide peptide exhibited rapid clearance from the blood circulation with low to moderate (up to 5% ID/g) uptake/retention by the major body organs. The accumulation in the estrogen receptor-positive MCF7 tumor was 2.24% ± 0.62% ID/g at 45 min p.i, with good tumor to blood and muscle uptake ratios. The radiolabeled peptide was excreted primarily through the renal pathway. Conclusion: The encouraging results of this initial study demonstrate that additional testing of this leuprolide peptide seems to be indicated because of its convincing potential to be a new agent for the management of breast carcinoma.