Recombinant targeted toxin based on HER2-specific DARPin possesses a strong selective cytotoxic effect in vitro and a potent antitumor activity in vivo

Evgeniya Sokolova; Galina Proshkina; Olga Kutova; Olga Shilova; Anastasiya Ryabova; Alexey Schulga; Oleg Stremovskiy; Tatiana Zdobnova; Irina Balalaeva; Sergey Deyev

doi:10.1016/j.jconrel.2016.05.020

Recombinant targeted toxin based on HER2-specific DARPin possesses a strong selective cytotoxic effect in vitro and a potent antitumor activity in vivo

J Control Release. 2016 Jul 10:233:48-56. doi: 10.1016/j.jconrel.2016.05.020. Epub 2016 May 10.

Affiliations

¹ M.M. Shemyakin and Y.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 16/10 Miklukho-Maklaya St., Moscow 117997, Russia; Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Ave., Nizhny Novgorod 603950, Russia.
² M.M. Shemyakin and Y.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 16/10 Miklukho-Maklaya St., Moscow 117997, Russia.
³ Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Ave., Nizhny Novgorod 603950, Russia.
⁴ Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilova St, Moscow 119991, Russia; National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe highway, Moscow 115409, Russia.
⁵ M.M. Shemyakin and Y.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 16/10 Miklukho-Maklaya St., Moscow 117997, Russia; Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Ave., Nizhny Novgorod 603950, Russia. Electronic address: deyev@ibch.ru.

PMID: 27178808
DOI: 10.1016/j.jconrel.2016.05.020

Abstract

DARPins fused with other proteins are promising non-immunoglobulin scaffolds for specific binding to target cells. In this study HER2-specific DARPin (DARPin_9-29) was used as a tumor-targeting moiety for the delivery of a cytotoxic agent - the fragment of Pseudomonas aeruginosa exotoxin A. It was determined that DARPin-PE40 possesses a considerable cytotoxic activity and induces apoptosis in HER2-positive cells. Cytotoxic effect of DARPin-PE40 strongly correlates with the HER2 expression level. The effect of intravenous administration of DARPin-PE40 was tested in the xenograft model of breast cancer. It was shown that treatment of animals with DARPin-PE40 caused strong and prolonged suppression of xenograft tumor growth.

Keywords: DARPin; HER2; PE40; Pseudomonas aeruginosa exotoxin A; Recombinant targeted toxin; Targeted therapy.

MeSH terms

ADP Ribose Transferases / administration & dosage*
ADP Ribose Transferases / therapeutic use
Animals
Antineoplastic Agents / administration & dosage*
Antineoplastic Agents / therapeutic use
Apoptosis / drug effects
Bacterial Toxins / administration & dosage*
Bacterial Toxins / therapeutic use
CHO Cells
Cell Line, Tumor
Cell Survival / drug effects
Cricetulus
Exotoxins / administration & dosage*
Exotoxins / therapeutic use
Female
Humans
Immunotoxins / administration & dosage*
Immunotoxins / therapeutic use
Mammary Neoplasms, Experimental / drug therapy*
Mammary Neoplasms, Experimental / metabolism
Mammary Neoplasms, Experimental / pathology
Mice, Nude
Pseudomonas aeruginosa Exotoxin A
Receptor, ErbB-2 / metabolism*
Recombinant Fusion Proteins / administration & dosage*
Recombinant Fusion Proteins / therapeutic use
Tumor Burden / drug effects
Virulence Factors / administration & dosage*
Virulence Factors / therapeutic use

Substances

Antineoplastic Agents
Bacterial Toxins
Exotoxins
Immunotoxins
Recombinant Fusion Proteins
Virulence Factors
ADP Ribose Transferases
ERBB2 protein, human
Receptor, ErbB-2