In vitro and in vivo Evaluation of the Radiochemical Compound Based on 99mTechnetium Labelled DARPin 9_29 for Molecular Visualization of Malignancies Overexpressing Her2/neu

O. D. Bragina; A. Vorobyeva; V. Tolmachev; A. Orlova; V. I. Chernov; S. Deyev; G. Proshkina; A. Shulga4; M. S. Lar&#039;kina; A. A. Medvedeva; R. V. Zel&#039;chan

doi:doi:10.12737/1024-6177-2020-65-1-37-41

UDC 61 CSCSTI 76.03 CSCSTI 76.33 Russian Classification of Professions by Education 14.04.02 Russian Classification of Professions by Education 31.06.2001 Russian Classification of Professions by Education 31.08.08 Russian Classification of Professions by Education 32.08.12 Russian Library and Bibliographic Classification 51 Russian Library and Bibliographic Classification 534 Russian Trade and Bibliographic Classification 5708 Russian Trade and Bibliographic Classification 5712 Russian Trade and Bibliographic Classification 5734 Russian Trade and Bibliographic Classification 6212

In vitro and in vivo Evaluation of the Radiochemical Compound Based on 99mTechnetium Labelled DARPin 9_29 for Molecular Visualization of Malignancies Overexpressing Her2/neu

Published в Medical Radiology and radiation safety · Volume 65, Issue 1, 2020 · Pages 37–41 · Rubrics: Radiation medicine

DOI 10.12737/1024-6177-2020-65-1-37-41

Received: 02.02.2020 Accepted: 09.02.2020 Published: 09.02.2020

Authors

O. D. Bragina ¹ · A. Vorobyeva ² · V. Tolmachev ³ · A. Orlova ⁴ · V. I. Chernov ⁵ · S. Deyev ⁶ · G. Proshkina ⁷ · A. Shulga4 ⁸ · M. S. Lar'kina ⁹ · A. A. Medvedeva ¹⁰ · R. V. Zel'chan ¹¹

1 Tomsk National Research Medical Center (Junior Researcher)
Tomsk, Russian Federation

2 Uppsala University (Researcher)

3 Uppsala University (Head of Laboratory)

4 Uppsala University (Head of Laboratory)

5 Tomsk National Research Medical Center (Head of Department)
, National Research Tomsk Polytechnic University (Deputy Director)
Tomsk, Russian Federation

6 National Research Tomsk Polytechnic University (Head of Laboratory)
, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Head of Laboratory)

7 Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Senior Researcher)

8 Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Senior Researcher)

9 Siberian State Medical University (kafedra farmacevticheskogo analiza, Associate Professor)
Tomsk, Russian Federation

10 Tomsk National Research Medical Center (Senior Researcher)
Tomsk, Russian Federation

11 Tomsk National Research Medical Center (Radiologist)
Tomsk, Russian Federation

ABSTRACT

Purpose: Evaluation of a radiopharmaceutical based on 99mTc-labeled targeted molecules DARPin9_29 for radionuclide diagnostics of malignancies with Her2/neu overexpression. Material and methods: The DARPin9_29 sequence was amplified from the plasmid pET-DARP-6HIS for the DARPin9_29-His6 gene expression in E. coli cells. The eluent of 99mTcO4– (400–500 μl, 4 GBq) was added to the kit and incubated at a temperature of 100 °C for 20 minutes. After incubation, 40 μl of tricarbonyl technetium was added to 168 μg of DARPin9_29 in 100 μl of PBS (sodium phosphate buffer), followed by incubation at 40 °C for 60 minutes. The radiochemical yield and purity were determined by thin layer radiochromatography, the purification was performed using NAP-5 cleansing columns (GE Healthcare). Cell lines with different levels of Her2/neu expression were used: SKOV-3> BT474 >> DU-145 for the determination of the radiopharmaceutical specificity. Her2/neu expressing cell line SKOV-3 was used for in vitro study. The study was conducted 6 hours after the administration of the drug. Results: The radiochemical yield was 72 ± 8 %, the radiochemical purity after purification was 98.7 ± 1.0 %. The stability in PBS (phosphate buffered saline) solution after 1 hour was 99.8 ± 0.2; after 3 hours – 98.2 ± 0.1. In vitro studies showed that the accumulation of explored compound was directly proportional to the level of Her2/neu expression in cells, while blocking the receptors with an excess of unlabeled protein showed a significant reduction in binding in the group of cells. Data on biodistribution and SPECT/CT in the body of the animal BALB/c nu/nu demonstrated rapid removal of the compound from the blood stream and high accumulation in the liver, kidney and bladder 6 hours after the introduction of the radiopharmaceutical. Conclusion: The studies demonstrated high radiochemical yields and purity, as well as stability of the studied compound. The results of in vitro and in vivo analysis showed the specificity and affinity of the radiopharmaceutical to the Her2/neu receptor on the surface of tumor cells. The high accumulation of the drug in the liver and kidneys, detected in in vivo studies, is probably due to the lipophilicity of the 99mTc(CO)3-histidine tag and indicates the limitation of its further clinical use in assessing the condition of the above organs, which will require additional diagnostic methods, as well as possible modification chemical structure.

KEYWORDS

malignancies Her2/neu radionuclide diagnostics DARPin9_29

Information References

Authors:

1. Tomsk National Research Medical Center (Junior Researcher)

Tomsk, Russian Federation

2. Uppsala University (Researcher)

3. Uppsala University (Head of Laboratory)

4. Uppsala University (Head of Laboratory)

5. Tomsk National Research Medical Center (Head of Department)

National Research Tomsk Polytechnic University (Deputy Director)

Tomsk, Russian Federation

6. National Research Tomsk Polytechnic University (Head of Laboratory)

Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Head of Laboratory)

7. Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Senior Researcher)

8. Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Senior Researcher)

9. Siberian State Medical University (kafedra farmacevticheskogo analiza, Associate Professor)

Tomsk, Russian Federation

10. Tomsk National Research Medical Center (Senior Researcher)

Tomsk, Russian Federation

11. Tomsk National Research Medical Center (Radiologist)

Tomsk, Russian Federation

Type:

Article

Pages:

from 37 to 41

Status:

Published

Subject area:

UDC 61
CSCSTI 76.03
CSCSTI 76.33
Russian Classification of Professions by Education 14.04.02
Russian Classification of Professions by Education 31.06.2001
Russian Classification of Professions by Education 31.08.08
Russian Classification of Professions by Education 32.08.12
Russian Library and Bibliographic Classification 51
Russian Library and Bibliographic Classification 534
Russian Trade and Bibliographic Classification 5708
Russian Trade and Bibliographic Classification 5712
Russian Trade and Bibliographic Classification 5734
Russian Trade and Bibliographic Classification 6212

Language:

Russian

Keywords:

malignancies, Her2/neu, radionuclide diagnostics, DARPin9_29

References

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In vitro and in vivo Evaluation of the Radiochemical Compound Based on 99mTechnetium Labelled DARPin 9_29 for Molecular Visualization of Malignancies Overexpressing Her2/neu

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