CS252230B1 - A method for the preparation of radioisotope-labeled D-glucosemin - Google Patents

Abstract

The essence of the proposed solution is that labeled 1-amino-1-deoxy-D-fructose or 1-N-alkyl/or aryl/amino-1-deoxy-D-fructose is reacted with liquid ammonia or methanolic ammonia or ethanolic ammonia, preferably at a temperature of 18 to 25 °C for 4 to 24 days in a closed system in the presence of ammonium chloride in an amount of 0.5 to 3 moles per 1 mole of starting material and/or at least one catalyst from the group including succinic acid, glutaric acid, benzoic acid, gallic acid, preferably in an amount of 3 to 8 moles per 1 mole of starting material.

Description

The present invention provides a process for the preparation of radiolabelled C-labeled D-glucosamine from 1-amino-1-deoxy-D-fructose or N-substituted derivatives thereof.

D-Glucosamine is an extremely important natural aminosaccharide whose occurrence is characteristic mainly for various types of polysaccharides, lipopolysaccharides and glycoproteins. Research into substances containing D-glucosamine is one of the key aspects of biochemistry. D-Glucosamine is observed mainly as a part of the skeleton of muramic and neuraminic acid, biologically very important, and in the form labeled with the carbon C isotope is of invaluable importance in this respect.

A process for the preparation of radiolabelled C-labeled D-glucosamine from D-fructose as a starting material is the subject of AO 231 498. D-Fructose, however, labeled carbon isotopes are more difficult to obtain than economically more advantageous labeled D-glucose, from which labeled D-fructose is also commonly obtained (isomerization catalysed by enzymes, etc.). It would therefore be advantageous to use directly D-glucose to prepare labeled D-glucosamine.

In the field of unlabeled compounds, a common source of D-glucosamine is natural substances, especially chitin / Stacey, Webber, J. M. Methods in Carbohyd. Chem. 2, 228 (1962).

The only available literature source discloses the preparation of D-glucosamine by reacting D-glucose with ammonia at 100 ° C (US Patent 2,884,441).

However, the reaction conditions are too vigorous to work with carbon-labeled D-glucose and there is a large loss of radioactivity in the degradation products, which is about 80%, while the yields of labeled D-glucosamine are low.

Using the Amadori rearrangement, 1-amino-1-deoxy-D-fructose can be prepared from D-glucose [Kuhn R., Haas, H. J., Liebig's Ann. Chem., 600, 148 (1956); Druey J. and Huber G .: Helv. Chim.

Acta 40, 342 (1957), which reacts with ammonia to provide D-glucosamine (U.S. Patent 2,918,462).

The formation of D-glucosamine in this reaction, which has not been theoretically studied, would probably be accounted for by the mechanism of Heyns rearrangement / Heyns K., Koch W .: Z. Naturforsch.

7b, 486 (1952), Heyns. , Meinecke, K. H., Chem. Ber. 86, 1453 (1953), The Carbohydrates (Pigman W., Horton D., Eds., Academic Press, New York 1980, 905), provided that the amino group of 1-amino-1-deoxy-D-fructose It would behave analogously to the C1-hydroxyl group of D-fructose (which, when reacted with ammonia, provides D-glucosamine. See, e.g., Heyns, K., Paulsen, H., Eichstedt, R., Rolle, M .: Chem. Ber. 20, 2039 (1957) and in the field of labeled compounds AO 231 498).

Reaction of derivative I with ammonia affords 1-amino-1-deoxy-D-fructosylamine II, further providing endiamine III to form tautomeric aldimine IV, readily hydrolyzable to D-glucosamine V;

HIM

OH _o c ^h 2nh ₂ ho}

OH

NH,

HIM

OH

HO / hoh ₂ c

CHNH,

II

C-NH,

OH

III

AND

HC

1 ° ^H CH = NH

NH,

IN

IV

Based on an analogous scheme, one would also expect that N-substituted 1-amino-1-deoxy-D-fructose derivatives, i.e. 1-N-alkyl (or aryl) amino-1-deoxy-D-fructose IV) R = primary alkyl or aryl (could be reacted with ammonia by way of Heyns rearrangement (see the above derivative of derivative II to aldimine IV) to provide N-substituted aldimine VII, again hydrolyzable to D-glucosamine V):

This type of ties has not been described in the available literature yet.

The starting derivatives are available from D-glucose and suitable amine types using Amadori rearrangement of the transient N-substituted D-glucosylamines / Hodge JE, Fisher BE: Methods Carbohyd. Chem. 2_ _f 99/1963 /, r Weygand .: Chem. Ber. 73 "1259 (1940), Micheel F., Hagemann G.: Chem. Ber. 9 3, 2 381 (1960), also in radioisotope-labeled form (Skála L .: Candidate dissertation, Faculty of Science, Charles University, Prague 1984, p. 36).

The formation of D-glucosamine in reactions of derivatives IV with ammonia and hydrolysis of the corresponding aldimines VII has been demonstrated and verified on ¹⁴ C-labeled and non-labeled substances.

The proposed process is based on the reaction principle. D-glucosamine 14 labeled with radioisotope C, characterized in that 1-amino-1-deoxy-D-fructose or 1-N-alkyl / or aryl / amino-1-deoxy-D-fructose is reacted with liquid ammonia or methanolic ammonia or ethanolic ammonia, preferably at 18 to 25 ° C for 4 to 24 days, in a closed system in the presence of 0.5 to 3 moles of ammonium chloride per mole of starting material and / or at least one catalyst from the group comprising succinic acid, glutaric acid, benzoic acid, gallic acid, preferably in an amount of 3 to 8 mol of the starting material.

The reaction may be carried out with liquid ammonia for 5 to 8 days in the presence of 1 to 3 moles of ammonium chloride per mole of the starting material or for 4 to 10 days in the presence of 1 to 2 moles of ammonium chloride and / or at least one catalyst selected from succinic acid, glutaric acid, benzoic acid and gallic acid in an amount of 4 to 8 moles per mole of starting material.

The reaction with liquid ammonia can also be carried out for 4 to 6 days in the presence of 0.5 to 2 moles of ammonium chloride per mole of starting material, then remove the liquid ammonia, add methanol or ethanol saturated at 0 ° C with ammonia gas and at least one catalyst from the group consisting of succinic acid, glutaric acid, benzoic acid and gallic acid in an amount of 4 to 6 moles per 1 mol of starting material and allowed to proceed for a further 10 to 12 days.

The reaction can also be carried out with methanolic or ethanolic ammonia prepared by saturating the corresponding alcohol at 0 ° C with ammonia gas for 20 to 24 days in the presence of ammonium chloride in an amount of 1 to 2 moles per 1 mol of starting material and / or at least one a catalyst selected from the group consisting of succinic acid, glutaric acid, benzoic acid and gallic acid in an amount of 3 to 6 moles of 1 mole of starting material. The acid catalyst can be added to the reaction mixture only after 8 to 12 days from the start of the reaction.

In all these cases, after completion of the reaction, ammonia is removed from the reaction mixture and the resulting aldimines, preferably diluted with hydrochloric acid, are hydrolyzed to D-glucosamine, which is isolated in the form of a salt, preferably as the hydrochloride.

The advantage of this method of preparing radioisotope-labeled D-glucosamine is that

SpOČÍvá

- D-glucosamine is prepared from an available and economically advantageous source, i.e. D-glucosine,

- D-glucose can be converted to starting materials by reaction with amines using Amadori rearrangement in yields of over 80

- the starting substances can be regenerated, so that up to 70% of the total radioactivity is recovered,

The reaction of 1-N-alkyl / or aryl / amino-1-deoxy-D-fructose with ammonia is a new general type of reaction allowing the choice of the 1-N-substituent.

The process according to the invention is further illustrated in the following examples.

Example 1 1-Amino-1-deoxy-D- (U-C) fructose) in the form of acetate (40 MBq); default activity

-1

D- (U-C) fructose 9.2 GBq.mmol, which also applies to Examples 2 to 6), 1 ml of liquid ammonia and 0.6 mg of ammonium chloride were reacted for 5 days at 20 to 22 ° C. C in a sealed glass ampoule. The ammonia is evaporated, 0.5 ml of 0.5 M hydrochloric acid is added and the solution is heated to 50 to 60 ° C for 5-10 min.

D- (U-C) Glucosamine in the hydrochloride form can be isolated, for example, by preparative paper chromatography in the 1-butanol-ethanol-water 52:33:15 system. The same applies further to Examples 14-6. The radiochemical yield of D- (U-C) glucosamine hydrochloride is 44%.

Example 2 1-N-Benzylamino-1-deoxy-D- (U-C) fructose (hydrochloride) (61 MBq), 1.5 ml of liquid ammonia, 1 mg of ammonium chloride and 3.5 mg of glutaric acid are reacted for 8 days in an autoclave at 23-25 ° C. The ammonia is evaporated, 2 ml are added

0.5M hydrochloric acid and the solution is heated to 60 ° C for 15-20 min. The radiochemical yield of D- (U-C) glucosamine hydrochloride is 22%.

Example 3 1-N-Cyclohexylamino-1-deoxy-D- (U-C) fructose] in the form of the hydrochloride (83 MBq),

1.5 ml of liquid ammonia and 0.5 mg of ammonium chloride are reacted in a sealed vial for 6 days.

The ammonia is evaporated, 2.5 mg of succinic acid, 2.7 mg of benzoic acid and 1.5 ml of anhydrous ethanol, saturated at 0 ° C with ammonia gas, are added and reacted in a sealed vial at the temperature of the first stage of the reaction, i.e., 20-25 ° C. The solution was evaporated in vacuo, followed by hydrolysis with hydrochloric acid as in Example 2. The radiochemical yield of D- (U-C) glucosamine hydrochloride was 19%.

Example 4 LN-phenylamino-l-deoxy-D / U- ¹⁴ C / fructose / hydrochloride / / 48 MBq / 4 mL of anhydrous methanol saturated at 0 ° C with gaseous ammonia, 0.5 mg of ammonium chloride and 7 1.8 mg of gallic acid is allowed to react in a well sealed flask at 22-25 ° C for 22 days.

The solution was evaporated in vacuo, 2.5 ml of 0.5 M hydrochloric acid was added and the solution was heated to 65 C for 14 min. For 25 min. The radiochemical yield of D- (U-C) glucosamine hydrochloride was 29%.

EXAMPLE 5 1 N-Tolylamino-1-deoxy-D- (U-C) fructose as hydrochloride (64 MBq), 4 ml of methanol saturated at 0 ° C with ammonia gas and 0.6 mg of ammonium chloride were reacted in well sealed flask at 20 to 25 ° C for 10 days.

A solution of 4.3 mg of succinic acid in 0.3 ml of methanol was then added to the reaction mixture and the reaction mixture was allowed to stand at the same temperature for a further 11 days. The solution was evaporated in vacuo and hydrolyzed with hydrochloric acid as in Example 4. The radiochemical yield of D- (U-C) glucosamine hydrochloride was 33%.

EXAMPLE 1β-Tolylamine-1,1'-deoxy-D- (U-C) fructose (hydrochloride) (39 MBq), 1.5 ml of liquid ammonia, 2.8 mg of succinic acid and 2.1 mg of glutaric acid are left to leave. react in a stopped vial at 22-25 ° C for 8 days.

The ammonia was evaporated and hydrolyzed with hydrochloric acid as in Example 4.

The radiochemical yield of D- (U-C) glucosamine hydrochloride is 15.5%.

Claims (4)

A process for the preparation of a radiolabelled D-glucosamine, characterized in that the labeled 1-amino-1-deoxy-D-fructose or 1-N-alkyl / or aryl / amino-1-deoxy-D-fructose is mentioned in reacting with liquid ammonia and / or methanolic ammonia and / or ethanolic ammonia, preferably at a temperature of 18 to 25 ° C for 4 to 24 days in a closed system in the presence of 0.5 to 3 moles of ammonium chloride per mole of starting material; and / or at least one catalyst selected from the group consisting of succinic acid, glutaric acid, benzoic acid and gallic acid, preferably in an amount of 3 to 8 moles per mole of starting material. 2. A process according to claim 1 wherein the reaction with liquid ammonia is carried out for 4 to 6 days in the presence of 0.5 to 2 moles of ammonium chloride per mole of starting material, after which the liquid ammonia is removed and methanol or ethanol is added. saturated at 0 ° C with ammonia gas and at least one catalyst selected from succinic acid, glutaric acid, benzoic acid and gallic acid in an amount of 4 to 6 moles per 1 mole of starting material and the mixture are allowed to react for a further 10 to 12 days. 3. The process of claim 1 wherein the reaction is carried out with methanolic or ethanolic ammonia prepared by saturating the corresponding alcohol at 0 [deg.] C. with ammonia gas for a period of time. 20 to 24 days, in the presence of ammonium chloride in an amount of 1 to 2 moles per 1 mol of starting material and / or at least one catalyst selected from succinic acid, glutaric acid, benzoic acid and gallic acid in an amount of 3 to 6 moles per 1 mol starting materials. 4. The process of claim 1 wherein the acid catalyst is added to the reaction mixture only 8 to 10 days after the start of the reaction.