Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates

David L. Kukis, Sally J. DeNardo, Gerald L. DeNardo, Robert T. O'Donnell, Claude F. Meares

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Abstract

Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10- tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) binds 90Y, with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y- DOTA immunoconjugate product yields have been typically only ≤50%. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. Methods: (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product. 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. Results: Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94% of 90Y, (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.6. Raising the reaction temperature from 25°C to 37°C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37°C. In eight labeling experiments under optimal conditions, a mean product yield (± s.d.) of 91% ± 8% was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y, per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100% ± 11%. Conclusion: The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals.

Original languageEnglish (US)
Pages (from-to)2105-2110
Number of pages6
JournalJournal of Nuclear Medicine
Volume39
Issue number12
StatePublished - Dec 1998

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Immunoconjugates
Yttrium
Radioimmunotherapy
Buffers
Reaction Time
Temperature
Halogenation
Chelating Agents
Monoclonal Antibodies
Clinical Trials
Costs and Cost Analysis
Bone and Bones
Peptides

Keywords

  • 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''- tetraacetic acid
  • Chelate
  • Radioimmunotherapy
  • Yttrium-90

ASJC Scopus subject areas

  • Radiological and Ultrasound Technology

Cite this

Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates. / Kukis, David L.; DeNardo, Sally J.; DeNardo, Gerald L.; O'Donnell, Robert T.; Meares, Claude F.

In: Journal of Nuclear Medicine, Vol. 39, No. 12, 12.1998, p. 2105-2110.

Research output: Contribution to journalArticle

Kukis, David L. ; DeNardo, Sally J. ; DeNardo, Gerald L. ; O'Donnell, Robert T. ; Meares, Claude F. / Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates. In: Journal of Nuclear Medicine. 1998 ; Vol. 39, No. 12. pp. 2105-2110.
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abstract = "Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10- tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) binds 90Y, with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y- DOTA immunoconjugate product yields have been typically only ≤50{\%}. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. Methods: (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product. 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. Results: Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94{\%} of 90Y, (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.6. Raising the reaction temperature from 25°C to 37°C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37°C. In eight labeling experiments under optimal conditions, a mean product yield (± s.d.) of 91{\%} ± 8{\%} was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y, per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100{\%} ± 11{\%}. Conclusion: The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals.",
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T1 - Optimized conditions for chelation of yttrium-90-DOTA immunoconjugates

AU - Kukis, David L.

AU - DeNardo, Sally J.

AU - DeNardo, Gerald L.

AU - O'Donnell, Robert T.

AU - Meares, Claude F.

PY - 1998/12

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N2 - Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10- tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) binds 90Y, with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y- DOTA immunoconjugate product yields have been typically only ≤50%. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. Methods: (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product. 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. Results: Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94% of 90Y, (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.6. Raising the reaction temperature from 25°C to 37°C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37°C. In eight labeling experiments under optimal conditions, a mean product yield (± s.d.) of 91% ± 8% was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y, per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100% ± 11%. Conclusion: The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals.

AB - Radioimmunotherapy (RIT) with 90Y-labeled immunoconjugates has shown promise in clinical trials. The macrocyclic chelating agent 1,4,7,10- tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) binds 90Y, with extraordinary stability, minimizing the toxicity of 90Y-DOTA immunoconjugates arising from loss of 90Y to bone. However, reported 90Y- DOTA immunoconjugate product yields have been typically only ≤50%. Improved yields are needed for RIT with 90Y-DOTA immunoconjugates to be practical. Methods: (S) 2-[p-(bromoacetamido)benzyl]-DOTA (BAD) was conjugated to the monoclonal antibody Lym-1 via 2-iminothiolane (2IT). The immunoconjugate product. 2IT-BAD-Lym-1, was labeled in excess yttrium in various buffers over a range of concentrations and pH. Kinetic studies were performed in selected buffers to estimate radiolabeling reaction times under prospective radiopharmacy labeling conditions. The effect of temperature on reaction kinetics was examined. Optimal radiolabeling conditions were identified and used in eight radiolabeling experiments with 2IT-BAD-Lym-1 and a second immunoconjugate, DOTA-peptide-chimeric L6, with 248-492 MBq (6.7-13.3 mCi) of 90Y. Results: Ammonium acetate buffer (0.5 M) was associated with the highest uptake of yttrium. On the basis of kinetic data, the time required to chelate 94% of 90Y, (four half-times) under prospective radiopharmacy labeling conditions in 0.5 M ammonium acetate was 17-148 min at pH 6.5, but it was only 1-10 min at pH 7.6. Raising the reaction temperature from 25°C to 37°C markedly increased the chelation rate. Optimal radiolabeling conditions were identified as: 30-min reaction time, 0.5 M ammonium acetate buffer, pH 7-7.5 and 37°C. In eight labeling experiments under optimal conditions, a mean product yield (± s.d.) of 91% ± 8% was achieved, comparable to iodination yields. The specific activity of final products was 74-130 MBq (2.0-3.5 mCi) of 90Y, per mg of monoclonal antibody. The immunoreactivity of 90Y-labeled immunoconjugates was 100% ± 11%. Conclusion: The optimization of 90Y-DOTA chelation conditions represents an important advance in 90Y RIT because it facilitates the dependable and cost-effective preparation of 90Y-DOTA pharmaceuticals.

KW - 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''- tetraacetic acid

KW - Chelate

KW - Radioimmunotherapy

KW - Yttrium-90

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