title: Tracer level radiochemistry to clinical dose preparation of (177)Lu-labeled cyclic RGD peptide dimer.
abstract: OBJECTIVE
  Integrin αvβ3 plays a significant role in angiogenesis during tumor growth and metastasis, and is a receptor for the extracellular matrix proteins with the exposed arginine(R)-glycine(G)-aspartic acid(D) tripeptide sequence. The over-expression of integrin αvβ3 during tumor growth and metastasis presents an interesting molecular target for both early detection and treatment of rapidly growing solid tumors. Considering the advantages of (177)Lu for targeted radiotherapy and enhanced tumor targeting capability of cyclic RGD peptide dimer, an attempt has been made to optimize the protocol for the preparation of clinical dose of (177)Lu labeled DOTA-E[c(RGDfK)]2 (E=Glutamic acid, f=phenyl alanine, K=lysine) as a potential agent for targeted tumor therapy.
  
  
  METHODS
  (177)Lu was produced by thermal neutron bombardment on enriched Lu2O3 (82% in (176)Lu) target at a flux of 1 × 10(14) n/cm(2).s for 21 d. Therapeutic dose of (177)Lu-DOTA-E[c(RGDfK)]2 (7.4GBq) was prepared by adding the aqueous solution of the ligand and (177)LuCl3 to 0.1M NH4OAC buffer containing gentisic acid and incubating the reaction mixture at 90°C for 30 min. The yield and radiochemical purity of the complex was determined by HPLC technique. Parameters, such as, ligand-to-metal ratio, pH of the reaction mixture, incubation time and temperature were varied using tracer quantity of (177)Lu (37 MBq) in order to arrive at the optimized protocol for the preparation of clinical dose. Biological behavior of the radiotracer prepared was studied in C57/BL6 mice bearing melanoma tumors.
  
  
  RESULTS
  (177)Lu was produced with a specific activity of 950 ± 50 GBq/mg (25.7 ± 1.4 Ci/mg) and radionuclidic purity of 99.98%. A careful optimization of several parameters showed that (177)Lu-DOTA-E[c(RGDfK)]2 could be prepared with adequately high radiochemical purity using a ligand-to-metal ratio ~2. Based on these studies therapeutic dose of the agent with 7.4 GBq of (177)Lu was formulated in ~63 GBq/μM specific activity with high yield (98.2 ± 0.7%), radiochemical purity and in vitro stability. Biodistribution studies carried out in C57/BL6 mice bearing melanoma tumors revealed specific accumulation of the radiolabeled conjugate in tumor (3.80 ± 0.55% ID/g at 30 min p.i.) with high tumor to blood and tumor to muscle ratios. However, the uptake of the radiotracer in the tumor was found to be reduced to 1.51 ± 0.32 %ID/g at 72 h p.i.
  
  
  CONCLUSIONS
  The present work successfully demonstrates the formulation of an optimized protocol for the preparation of (177)Lu labeled DOTA-E[c(RGDfK)]2 for PRRT applications using (177)Lu produced by direct neutron activation in a medium flux research reactor.
authors.affiliation: Radiopharmaceuticals Division, Bhabha Atomic Research Centre, Mumbai 400085, India. Electronic address: sudipta@barc.gov.in.
authors.forename: Sudipta
authors.initials: S
authors.lastname: Chakraborty
authors.forename: H D
authors.initials: HD
authors.lastname: Sarma
authors.forename: K V
authors.initials: KV
authors.lastname: Vimalnath
authors.forename: M R A
authors.initials: MR
authors.lastname: Pillai
mesh_terms: D000818:Animals; D019281:Dimerization; D004355:Drug Stability; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006801:Humans; D007553:Isotope Labeling; D008187:Lutetium / Q000627:therapeutic use*; D008545:Melanoma / Q000532:radiotherapy; D051379:Mice; D000442:Octanols / Q000737:chemistry; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000493:pharmacokinetics / Q000627:therapeutic use*; D011829:Radiation Dosage; D011849:Radioactive Tracers; D011854:Radiochemistry / Q000379:methods*; D011868:Radioisotopes / Q000627:therapeutic use*; D014867:Water / Q000737:chemistry
chemical_list: D006573:Heterocyclic Compounds, 1-Ring; D000442:Octanols; D010456:Peptides, Cyclic; D011849:Radioactive Tracers; D011868:Radioisotopes; C507169:cyclic arginine-glycine-aspartic acid peptide; D014867:Water; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; D008187:Lutetium
keywords: (177)Lu; Clinical dose; Integrin α(v)β(3); RGD peptide; Targeted tumor therapy