RU2233285C1 - Highly tritium labeled [3h]-rapamycin - Google Patents

Abstract

FIELD: organic chemistry, labeled compounds.

SUBSTANCE: invention relates to a new tritium-labeled analogue of physiologically active compound, namely [³H]-rapamycin of the formula (I):

that is used in organic and bioorganic chemistry and in medicine as an immunosuppressor. Invention provides preparing analogue of physiologically active compound.

EFFECT: valuable properties of compound.

1 cl, 1 ex

Description

The invention relates to organic chemistry and can find application in analytical chemistry, bioorganic chemistry, biochemistry and applied medicine.

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

Known rapamycin formula I

This compound, called rapamycin, is a powerful immunosuppressant and reagent used in immunochemistry (NHSigal, FJDumont // Ann.Rev.Immunol., 1992. Vol.10. 519-560. JYChang, SNSehgai // Brit.J. Rheumatol. 1991. Vol. 30. No. 2. 62-65) [1].

However, its tritium-labeled analog is not described.

It is known that the replacement of 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) [2].

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 highly labeled tritium ³ H-rapamycin of the formula I.

The following is an example implementation of the invention.

Example

In the first chamber of the reaction two-chamber ampoule with the first lower and second upper chambers communicating with each other, 40 mg of palladium oxide and 40 mg of 5% PdO / BaSO _{4 were} placed, and 50 μl of abs. dioxane, 5 μl of triethylamine and 12 mg of rapamycin. The second chamber was frozen with liquid nitrogen and the ampoule was evacuated to a pressure of 0.1 Pa and filled with gaseous tritium to a pressure of 333 GPa. Then the first chamber was heated to 70 ° C. Palladium oxide was reduced, tritium water was frozen in the second chamber. The reaction vial was evacuated to a pressure of 0.1 Pa, continuing to heat the first vial to 70 ° C, and filled with argon. Then the contents of the second chamber were transferred to the first chamber, which was sealed. Thus, the reaction mixture consisted of a reduced catalyst, 100% tritium water, triethylamine and a solution of rapamycin in dioxane. The contents of the ampoule were heated for 30 min at 100 ° C, after which the ampoule was opened, the reaction mixture was diluted with 0.5 ml of chloroform. The catalyst was filtered off and washed successively with chloroform (3 × 1 ml) and methanol (3 × 1 ml). Tritium water, dioxane and triethylamine were distilled off. Labile tritium was removed by dissolving the substance several times in methanol (5 × 2 ml) and evaporating the latter.

Purification of the labeled preparation was carried out by HPLC in two stages:

1) on a Kromasil 100C ₁₈ , 7 μm column (8 × 250 mm), in a 30-minute gradient from 0 to 100% eluent B in eluent A, where eluent A is methanol-water (70:30), eluent B is methanol ; eluent consumption - 2 ml / min; retention time of rapamycin - 17.78 min;

2) on a Kromasil 100C column ₁₈ , 7 μm (8 × 250 mm), in a 30-minute gradient from 0 to 100% eluent C in eluent D, where eluent C is acetonitrile-water (60:40), eluent D is acetonitrile ; eluent consumption - 2 ml / min; retention time of rapamycin - 17.70 min,

The yield of tritium-labeled [ ³ H] -rapamycin after chromatography was 15-20%, molar radioactivity was 4.6 Ci / mmol.

Thus, a new physiologically active compound highly labeled with tritium was obtained.

Claims (1)

Highly labeled tritium [ ³ H] -rapamycin of the formula