production and quality control of 177lu-dotatate …
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PRODUCTION and QUALITY CONTROL of 177Lu-DOTATATE [177Lu-DOTA-Tyr3]- OCTREOTATE: CLINICAL APPLICATION
M.F. de Barboza; R. Herrerias; A.A. de Souza; G. Pereira; J.A. Pires; N. T.O. Fukumori; M.M.N. Matsuda; E. V.
Almeida; J. Mengatti; A. J. Belfer and L.N. Hilario
Radiopharmacy
Directory, IPEN-CNEN/SP
Diagnósticos Médicos, Nuclear Clinica de Radioisótopos, São Paulo -
BrazilIEA2009
INTRODUCTION
Somatostatin receptors have been identified in different kinds of tumors: neuroendocrine tumors and tumors of central nervous system, breast, lung and lymphatic tissue, making these receptors potential targets for radionuclide diagnostics and therapy.
Somatostatin receptors have served as the biomolecularbasis for the clinical use of radiolabeled somatostatinanalogues which, at present, are of great interest in nuclear medicine for diagnostic and peptide receptor radionuclide therapy (PRRT) applications.
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There are few treatment modalities for metastasized (GEP) neuroendocrine gastroentero - pancreatic tumors:
surgery,
(chemo)- embolization,
chemotherapy,
treatment with somatostatine (SST) analogous,
peptide receptor radionuclide therapy (PRRT) offers therapeutic strategy because a majority of GEP tumors possesses somatostatine receptors (SSTRs).
INTRODUCTION
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INTRODUCTION
Several radiolabeled somatostatin analogues are currently used, such a octreotide or octreotate, a complexing moity and a radionuclideThe basic principle envolves binding SSTRs, which are expressed on the cell surface of the tumor cell, followed by internalization of the radionuclide-peptide complexRadionuclides in PRRT:
β- 131I, 90Y, 186Re, 188Re, 177Lu
α 211At, 213Bi, 225Ac
Electron Auger 111In, 123I, 125I, 67Ga
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177Lu; 68Ga; 90Y; 111In
DOTA-DPhe1-Tyr3-Octreotate ⇒ DOTATATE
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Lu-177 - phisical and chemical characterics half life = 6.7 dayslow β-
energy = 149 -
497 keVγ
= 113 –
208 keV 11% (for imaging)
OBJECTIVE
The aim of this work was to present the production and the quality control of 177Lu-Tyr3-octreotate (177Lu-DOTATATE), using DOTA (1,4,7,10-tetrazacyclododecane-N,N´,N”,N´”-tetra acetic acid) as chelating agent.
The process was developed and validated in a “hot-cell” under cGMP conditions, at the Radiopharmacy Directory, IPEN-CNEN/SP.
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The labeling was performed in a “hot-cell”177LuCl3 (Lutetium trichloride) IDB- Holland
specific activity = 20 –
25 Ci
/ mg
DOTATATE “kit” containing:(0.4 mg DOTATATE; 42 mg gentisic
acid and 210
mg ascorbic acid/mL
sodium acetate buffer pH 4.5)Molar relation: peptide:radionuclide
(2:1)
Reaction: 30 minutes at 82-83 ºC After cooling: 1 mL of DTPA solution (4 mg DTPA / 3
mL 0.9% NaCl) was added
Material - Method
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The volume was completed until desirable radioactive concentration, approximately 7,400 MBq / mL 0.9% sodium chloride solution, under aseptic conditions
Sterilization through 0.22 μm Millipore filter
The doses were fractioned according to demand in sterile vials
CLINICAL APPLICATION “Albert Einstein Hospital”Dose: 7,400MBq / 1 –
2mL (vial) Dr. Aron
Belfer
Material - Method
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Radiopharmacy: “hot-cel”
Laminar Flow – Grade A
Eppendorf – dispenser
Dose calibrator - Capintec
“in side”
“in front”Glass vial
Lead Pot
Bucket
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Quality Control
Radiochemical purity
Determined by ITLC-SG (Instant Thin Layer Chromatography-Silica Gel)
0.1 mol/L sodium citrate, pH 5.5, as solvent
Retention factor (Rf ): 177Lu-Dotatate = 0.1- 0.2 177Lu3+ = 1.0
Determined by C18 Sep-Pak cartridge (Waters)177Lu3+ eluted with 5 mL 0.1 mol/L acetate buffer pH 5.5 177Lu-Dotatate eluted with 5 mL MeOH
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HPLC (LC-20AT Prominence) - Shimadzu, Japan
composed by two pumps, autosampler,
system controller and radiometric detector (Bioscan)
Column: Shim-Pack VP-ODS column (250 x 4.6 mm, 5 µm), at room temperature Volume: 10 µL Flow rate: 1.0 mL/min Gradient: A: water (0.1 % TFA); B: acetonitrile (0.1% TFA); 15%
B for 3 minutes; 15 to 50% B in 13 minutes; 50% B for 10 minutes; 50 to 70% B in 3 minutesTime: 30 minutes
Quality Control
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Sterility and pyrogen tests were performed by the microbiology procedures in different culture media and the “in-vitro” Limulus (LAL) test, respectively.
Quality Control
Culture Media
LAL
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Production documents IEA2009
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RESULTS
TIME (hours) 177Lu-DOTATATE
0.5 99.55 ±
0.02
4 99.53 ±
0.02
24 99.30 ±
0.05
48 99.41 ±
0.03
72 99.22 ±
0.07
Radiochemical purity of Radiochemical purity of 177177LuLu--DOTATATEDOTATATE
in 28 BatchesThe “hot cell” environment is monitored (viable and non viable particles) weekly by the Quality Control group.
All the equipment are calibrated.IEA2009
HPLC analysis showed labeling average efficiency of (99.98 ± 0.09)% and retention time (Rt) of the product in 14.74 minutes.
RESULTS
0.0 5.0 10.0 15.0 20.0 25.0 30.0
5.0
7.5
10.0
12.5
15.0
mV
B.Conc.(AD1
14.7
4Retention time (min)
AU
HPLC of HPLC of 177177LuLu--DOTATATEDOTATATE
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RESULTS
pH Purity (%) SP (MBq / mg) Sterility Pyrogen
4.5 99.55 ± 0.02 28.38 ± 4.70 negative negative
Characteristic of Characteristic of 177177LuLu--DOTATATEDOTATATE
Labeled with (45,732 ± 3,574)MBq of 177LuCl3 (IDB)
Stability of 177Lu-DOTA-Octreotate was high even 72 hours under refrigeration
Radiochemical purity >99.50% determined by ITLC-SG assay
Sterility and pyrogen tests were negative in all delivered vials
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CLINICAL APPLICATION
4 cycles 177Lu-Dotatate
Interval: between 6 and 10 weeks
Granisetron 1mg I.V.
Aminoacids I.V. in 4 h177Lu-Dotatate I.V. in 30 min
Hospitalization 24 h
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Case 1: Case 1: Pancreatic well diferentiated NETPancreatic well diferentiated NET
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Case 1: Case 1: Pancreatic well diferentiated NETPancreatic well diferentiated NET
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Case 2: Case 2: Pancreatic well diferentiated NETPancreatic well diferentiated NET
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In 28 batches / first semester of 2009:distributed: 511,780 MBq 177Lu-Dotatatedoses: 83specific activity: 28.38 ± 4.70MBq/µg
The labeling process in “hot-cell” and quality control procedures under cGMP conditions, have been successfully developed and validated at the Radiopharmacy Directory of IPEN–CNEN/SP.
Using the suitable methodology it is possible to provide high quality peptide RF for clinical use for PRRT in Brazil.
Conclusion
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19
9 10
16
87
14
6
17
12
4
0
5
10
15
20
jan fev mar apr may jun jul ago sep oct nov
Doses / month - 2009
120.6
59.666.8
96.9
52.044.6
75.3
44.4
111.6
67.7
15.5
0.020.040.060.080.0
100.0120.0140.0
jan fev mar apr may jun jul ago sep oct nov
MBq(103)/month - 2009
Distribution of 177Lu-Dotatate during 2009
Doses = 122
Total Activity = 754,985MBq
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