Methods of Assessing Biological Activity of Recombinant Human Interleukin-7 and Stability Study of the Preparation on Its Basis

Тетяна Миколаївна Луценко, Дарина Борисівна Старосила, Світлана Леонтіївна Рибалко, Олександр Юрійович Галкін

Abstract


Background. Interleukin-7 is one of the most important immune regulatory cytokines. Recombinant human interleukin-7 (rIL-7) in aqueous solution is subjected to chemical degradation mechanisms such as proteolysis, oxidation, disulfide exchange, oligomerisation etc. Such changes affect the shelf life of the preparation on the basis of rIL-7. Evaluation of the biological activity of rIL-7 can be carried out by assessing its antiviral activity.

Objective. Comparison of methods for inhibiting reproduction of the influenza virus, herpes simplex virus and hepatitis C virus with recombinant human interleukin-7 and research stability of the preparation on the basis of rIL-7.

Methods. We used immortalized cells: the kidneys of dogs, the bovine kidneys and kidneys of African green monkey Vero. The following viruses were used: hepatitis C virus surrogate (bovine viral diarrhea virus, BVDV), influenza virus (strain A/FM/1/47 (H1N1)), herpes simplex virus type 2 (HSV-2) (BH strain). To determine the antiviral activity of rIL-7 in vitro conditions using daily, immortalized cells. Cells were grown in RPMI-1640 medium.

Results. It was shown that rIL-7 in buffer stabilizing solutions and the culture medium after 1 week of storage was active at a dilution of 0.003 g/ml against BVDV. The study results of antiviral activity of rIL-7 drug in buffer solutions and culture medium against influenza virus A/FM/H1N1 can conclude that storage of the drug in buffer solution and in the intact state at 4 °C for one week has not affected its antiviral activity. It was shown that antiherpetic activity of preparations after 1 week of storage at 4 °C in buffer stabilizing solutions and intact state remained effective. The antiviral activity of drugs in a stabilizing solution has been persisted for 3 months at 4 °C, and in the intact state of rIL-7 lost its antiviral activity after 1 week against the herpes virus, and after 1 month regarding BVDV.

Conclusions. Methods for assessing the antiviral activity of rIL-7 towards BVDV, influenza virus, and HSV-2 were developed. It has been proven that the stabilizing buffer solutions proposed by us provide a high level of biological activity of rIL-7 preparations during storage at 4 °C for 3 months, which is a prerequisite for the development of liquid dosage forms of pharmaceutical preparations on their basis.

Keywords


Interleukin-7; Human recombinant; Antiviral activity; Biological standardization

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DOI: http://dx.doi.org/10.20535/1810-0546.2016.3.65102

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