RU2835090C1 - Deuterium-labelled boc-trp-pro-pro-trp - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: invention relates to organic chemistry and can be used in analytical chemistry and biological research. Disclosed is a novel deuterium-labelled Boc-Trp-Pro-Pro-Trp.

EFFECT: wider range of labelled analogues of physiologically active compounds.

1 cl, 1 dwg, 3 ex

Description

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

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

It is known that replacing atoms of compounds with their labeled analogues does not lead to changes in any properties of the original compound (Evans E.A. - Tritium and its compounds London Butterworths, 1974, p.48).

The original peptide Boc-Trp-Pro-Pro-Trp (Shevchenko KB, Andreeva LA, Nagaev IY, Myasoedov NF "Stability of the Boc-Trp-Pro-Pro-Trp peptide analogue PAM-43" // Abstract for the conference 6th Russian Conference on Medicinal Chemistry Medkhim 2024, Russia, Nizhny Novgorod, July 1-4, 2024, p. 375) was tested as an analogue of the positive allosteric modulator PAM-43 3,7-bis (2,3-dihydro-1-benzofuran-5-ylcarbonyl) -1,5-dimethyl-3,7-diazabicyclo [3.3.1] nonan-9-one, which can be used to treat Alzheimer's disease. Studies of the effect of the Boc-Trp-Pro-Pro-Trp peptide on the specific binding of hydrogen isotope-labeled [ ³ H]PAM-43 showed that the peptide affects the nature of specific binding of the radioligand, and the effect was detected both for the high-affinity binding site of PAM-43 and for its low-affinity site. This suggests the existence of targeted allosteric effects of the developed peptide on the glutamate receptor system of the mammalian brain, and also opens up prospects for further studies of the biological effects of this peptide, with the aim of creating drugs based on it in the future (Vyunova T.V., Shevchenko K.V., Nagaev I.Yu., Myasoedov N.F. "Assessment of the specific binding of Boc-Trp-Pro-Pro-Trp and Trp-Pro-Pro-Trp to plasma membrane cells of the rat brain" // Abstract for the conference 6th Russian Conference on Medicinal Chemistry Medkhim 2024, Russia, Nizhny Novgorod, July 1-4, 2024, p. 327).

Obtaining a deuterated analogue of Boc-Trp-Pro-Pro-Trp will thus allow for mass spectrometric analysis in in vivo experiments, which will accelerate the overcoming of the necessary stages for introducing this peptide into practical medicine as a means of stimulating brain function, facilitating learning, and improving memory.

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

The specified technical result is achieved by obtaining deuterium-labeled Boc-Trp-Pro-Pro-Trp.

Below are examples of the invention's implementation.

Example I

A solution of 10 mg Boc-Trp-Pro-Pro-Trp in 0.1 ml methanol was added to 400 mg 5% PdO/BaSO ₄ . Methanol was removed by evaporation on a rotary evaporator and the residue was lyophilized. Then 100 mg of the mixture was placed in an ampoule, which was evacuated to a pressure of 0.1 Pa, filled with gaseous deuterium to a pressure of 400 hPa and kept at a temperature of 170°C for 10 min. Excess gaseous deuterium was removed by evacuation. The substance was extracted from the catalyst with methanol (5x2 ml) and separated by filtration. Labile deuterium was removed several times by dissolving the substance in methanol (5x2 ml) and evaporating the latter. The yield of [T)]peptide was 16%, the average deuterium incorporation was 7.6 atoms. The analysis was performed on a Milichrom A-02 chromatograph, ProntoSIL-120-5-C18 AQ column (2×75 mm, particle size 5 μm), column temperature 35°C, wavelength 210 nm. Eluent A - _H2O +0.1% acetic acid, eluent B - methanol, linear gradient from 0 to 50% B in 10 min. Eluent feed rate 0.2 ml/min. Mass spectrometric data were obtained on an LCQ Advantage MAX instrument (TermoElektron, USA), with electrospray ionization, direct injection of a sample solution with a concentration of 10 μg/ml in water with the addition of 0.1% acetic acid and subsequent fragmentation of the molecular peak in the analyzer by the ion collision method at 35 eV.

Example II

Using a peptide-5% PdO/BaSO ₄ (1:40) mixture, isotope exchange with deuterium water was performed. 100 mg of the mixture was placed in an ampoule and treated with gaseous deuterium (400 hPa pressure) for 2 hours at room temperature. Then 0.1 ml of deuterium water was added to the ampoule and deuterium was displaced with argon. The ampoule was sealed and heated at 125°C for 10 minutes. The deuterium incorporation was 8.23 atoms per molecule, the yield of Boc-Trp-Pro-Pro-Trp was about 6%.

Example III

The synthesis was carried out using D ₂ and a mixture of peptide-5% PdO/BaSO4 (1:40) at a temperature of 150°C. 200 mg of a mixture of Boc-Trp-Pro-Pro-Trp and 5% PdO/BaSO ₄ were taken into the reaction, the gaseous deuterium pressure was 400 hPa. The reaction was carried out for 10 minutes at 150°C. Then the substance was washed off the catalyst with methanol (5x1 ml). The yield of the preparation was 25-30%, the deuterium content was within 4-5 atoms per molecule.

Thus, a new deuterium-labeled compound was obtained (see Fig. 1).

Claims (2)

Deuterium-labeled Boc-Trp-Pro-Pro-Trp.