Toxicity Study of Pharmacological Pair Encapsulated Citrobacter freundii C115H Methionine γ-Lyase / Methiin

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Abstract

The acute and subchronic toxicity of the pharmacological pair based on encapsulated Citrobacter freundii C115H methionine γ-lyase enzyme/prodrug (methiin) was studied in female ICR mice. The drug showed a weak/moderate dose-dependent hepatotoxic effect. Most of the identified changes in liver morphology were insignificant or mild deviations from the norm. Long-term use of a single therapeutic dose per mouse of 1.5 U C. freundii C115H methionine γ-lyase @ (PEG−P(Asp)70/PLL70)-PICsome / 2 mg methiin led to a slight decrease in the weight of animals without obvious signs of intoxication. A quarter of the animals in this group had no deviations from the norm in liver morphology. No nephrotoxic effect in all study groups was found.

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About the authors

S. V. Revtovich

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Author for correspondence.
Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

V. V. Kulikova

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

V. S. Koval

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

A. D. Lyfenko

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

V. A. Kazakov

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Pushchino, Moscow Region, 142290

A. S. Chernov

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Pushchino, Moscow Region, 142290

G. B. Telegin

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Pushchino, Moscow Region, 142290

A. S. Zemskaya

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

N. V. Anufrieva

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

E. A. Morozova

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

P. N. Solyev

Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences

Email: svetla21@mail.ru
Russian Federation, Moscow, 119991

References

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Supplementary files

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2. Appendix
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3. Fig. 1. Principle of action of the pharmacological pair encapsulated MGL/methionine.

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4. Fig. 2. Liver fragments of female mice of the control group. Haematoxylin and eosin staining. The arrows indicate: a - small foci of mononuclear infiltration in the parenchyma of one of the lobes of the organ (mouse 5.1); b - hypertrophy of individual hepatocytes (mouse 5.2).

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5. Fig. 3. Liver fragments of female mice on day 7 after a single injection of 1.5 U C115H-PIC-som MGL/2 mg methiin. Haematoxylin and eosin staining. The arrows indicate: a - small foci of mononuclear infiltration in the parenchyma of one of the organ lobes (mouse 1.2); b - hepatocyte karyomegaly (mouse 1.3).

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6. Fig. 4. Fragments of the liver of female mice on day 7 after a single administration of 1.5 units of C115H-PIC-com MGL / 10 mg of methiine. Staining with hematoxylin and eosin. The arrows indicate: a – small foci of infiltration by segmented leukocytes (mouse 2.1), b – foci of mononuclear infiltration with single segmented leukocytes and phagocytosis of dead hepatocytes (mouse 2.6).

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7. Fig. 5. Fragments of the liver of female mice on day 7 after a single administration of 1.5 units of C115H-PIC-com MG/20 mg of methiine. Staining with hematoxylin and eosin. The arrows indicate: a – the focus of hepatocytes with initial necrosis (mouse 3.4), b – hepatocytes with pigmented inclusions in the cytoplasm (mouse 3.5).

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8. Fig. 6. Fragments of the cortical (a) and cerebral (b) substances of the kidney of a female mouse of the control group (mouse 5.1). The usual histological structure of the organ. Staining with hematoxylin and eosin.

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9. Fig. 7. Fragments of the renal cortex of female mice on day 7 after a single administration of 1.5 units of C115H-PIC-com MG/2 mg of methiine. Staining with hematoxylin and eosin. The arrows indicate: a – the phenomena of chronic progressive nephropathy (mouse 1.5), b – a tubular cyst (solid arrow) lined with flattened epithelium (dotted arrows; mouse 1.3).

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10. Fig. 8. Fragments of the renal cortex of a mouse (3.5 mouse) on day 7 after a single administration of 1.5 units of C115H-PIC-com MG/20 mg of methiine. Staining with hematoxylin and eosin. The arrows indicate the phenomena of chronic progressive nephropathy (a) and perivascular focus of mononuclear infiltration (b).

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11. Fig. 9. Fragments of the liver of female mice of the control group. Staining with hematoxylin and eosin. a – The arrows indicate the focus of mononuclear infiltration with segmented leukocytes (mouse 5.4). b – The usual histological structure of the organ (mouse 5.3).

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12. Fig. 10. Fragments of the liver of female mice on day 15 after 7-fold administration of 1.5 units of C115H-PIC-com MG/2 mg of methiine. Staining with hematoxylin and eosin. The arrows indicate: a – a focus of hepatocytes with initial necrosis (mouse 4.1); b – hepatocytes with pigmented inclusions in the cytoplasm (mouse 4.3); c – small foci of extramedullary hematopoiesis (mouse 4.6); d – the usual histological structure of the organ (mouse 4.7).

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13. Fig. 11. Fragments of the cortical (a) and cerebral (b) substances of the mouse kidney on day 15 after 7-fold administration of 1.5 units of C115H-PIC-com MG/2 mg of methiine (mouse 4.4). The usual histological structure of the organ. Staining with hematoxylin and eosin.

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