RU2513852C1 - UNIFORMLY TRITIUM-LABELLED PYRO-Glu-His-Pro-NH2 - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to uniformly tritium-labelled pyro-Glu-His-Pro-NH₂, which can be used in analytical chemistry and biological research.

EFFECT: high efficiency of using the compound.

1 ex

Description

The invention relates to the field of organic chemistry and may find application in analytical chemistry and biological research.

When studying physiologically active compounds, their labeled analogues are necessary.

It is known that replacing the atoms of compounds with their labeled analogues does not lead to a change in any properties of the starting compound (Evans EA - Tritium and its compounds London Butterworths, 1974, p. 48) [1].

Known pyro-Glu-His-Pro-NH ₂ formula:

pyro-Glu-His-Pro-NH ₂ is the peptide hormone of the hypothalamus (E.A. Nillni, K.A. Sevarino. // Endocrine Reviews, 1999, V. 20, No. 5, P.599-648 [2]; R. O'Leary, B. O'Connor. // J. Neurochem., 1995, V. 65, P.953-963 [3]; N. Shibusawa, K. Hashimoto, M. Yamada. // The Cerebellum 2008, P.84-95 [4]; J. Boler, F. Enzmann, K. Folkers, CY Bowers, AV Serially. // Biochem. Biophys. Res. Commun., 1969, V. 37, P. 705 -710 [5]; V. Monga, CL Meena, N. Kaur, R. Jain. // Current Medicinal Chemistry, 2008, V. 15, P.2718-2733 [6]). This hormone is a neuropeptide, it takes part in the normalization of certain mental functions of the human body. For example, the presence of the antidepressant action of exogenous pyro-Glu-His-Pro-NH ₂ in depression was established. The main chemical and biological function of this hormone is to increase the production of prolactin and, to a greater extent, to increase the production of thyroid-stimulating hormone by the anterior pituitary gland.

However, its counterpart uniformly labeled with tritium is not described.

The technical result achieved by the present invention is to expand the range of labeled analogues of physiologically active compounds.

This technical result is achieved by obtaining uniformly labeled with tritium pyro-Glu-His-Pro-NH ₂ formula I:

The following is an example implementation of the invention.

Example I

A solution of 5 mg of pyro-Glu-His-Pro-NH ₂ in 75 μl of methanol was impregnated with 150 mg of aluminum oxide and lyophilized. The dried residue was mechanically mixed with 10 mg of 5% Pd / Al ₂ O ₃ . 66 mg of the mixture was placed in an ampoule, evacuated, and filled with gaseous tritium to a pressure of 400 hPa. The reaction was carried out at 200 ° C for 15 minutes. The ampoule was again evacuated, the catalyst was applied to the filter, the substance was extracted with methanol (5 × 0.5 ml). The extracts were evaporated, the residue was dissolved in methanol (3 × 1 ml) and again evaporated to remove labile tritium. The final methanol solution contained 180 mCi of peptide.

The analysis was performed on a Reprosil pur C18aq column, 4 * 150 mm, 5 μm, flow rate - 1 ml / min, A - water with 0.1% heptafluorobutyric acid, B - methanol, gradient B → 0-30% for 15 min: retention time pyro-Glu-His-Pro-NH ₂ - 11.54 min. Uniform pyro-Glu-His-Pro-NH ₂ was purified by preparative high performance liquid chromatography.

Chromatography was performed on a Kromasil 100C ₁₈ , 8 × 150 mm, 7 μm column, system: A — water with 0.1% heptafluorobutyric acid, B — methanol, gradient B → 0-50% in 30 min, flow rate - 2 ml / min. As a result, a labeled peptide with 75% radiochemical purity (80 mCi) was obtained.

After repeated purification, the radiochemical purity of the uniformly labeled pyro-Glu-His-Pro-NH ₂ is at least 98%, the yield is 15-20%, the molar radioactivity is 75 Ci / mmol.

Thus, pyro-Glu-His-Pro-NH ₂ uniformly labeled with tritium was obtained.

Claims (1)

Uniformly tritiated pyro-Glu-His-Pro-NH _{2 of the} formula: