CS253779B1 - Admantylamid of n-acetylmuramyl-l-alanyl-d-isoglutamine - Google Patents

Abstract

Said peptide substance is prepared by condensation of adamantylamide L-alanyl- -D-isoglutamine with protected muramic acid. They are removed from the protected glycopeptide protecting groups with, for example, sodium in liquid ammonia or by hydrogenation. This the new substance shows significant immunoadjuvant and immunostimulatory activity.

Description

The present invention relates to N-acetylmuramyl-L-alanyl-D-isoglutamine adamantylamide of the formula

CH ₂ - OH

HIM

/ in H, OH

NH-Acch ₃ -ch -CO-NH-CH-CO-NH -ch-co-nh ₂

CO - NH - Ad where Ad represents the residue of the adamantane bound at the 1-position and Ac represents the residue of acetic acid. These novel substances (hereinafter referred to as MAdDP brevity) have significant immunoadjuvant and immunostimulatory activity.

As is well known, there is still a lack of substances capable of enhancing the body's defenses by stimulating immune processes. Bacterial products or cell wall fragments have been used for some time for this purpose. Their disadvantage was the non-standard effects and serious side effects in clinical use. Similarly, in the field of substances with immunoadjuvant activity, in particular bacterial products from Mycobacterium tuberculosis are again used, often in combination with mineral oils.

Some clinical experience with the use of immunoadjuvant and immunostimulatory effects is currently known. Most experience has been gained with the known muramyldipeptide contained in bacterial wall fragments, the peptide contained in bacterial wall fragments, N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP), which has an immunoadjuvant but also pyrogenic effect and many other unfavorable properties for practice (A. Adam, E. Lederer, Med. Res. Rewiew 4, 111-152 (1984)).

This has led to the preparation of a number of synthetic analogues, some of which have a more selective pharmacological profile of effects, such as N-acetylmuramyl-L-alanyl-D-glutamine-n-butyl ester 2 (murabutide) according to P. Lefrancier, M. Derrien, X. Jamet , J. Chvay et al., J. Med. Chem. 25, (1982) or MDP-L-alanyl-phosphatidylethanolamine according to ³ J. Fidler, S. Sona, W. Vogler,

D. Smith, D. Braun, Tarosay L., Giesler R. and Sohroit A., Biol. Resp. Modif. 1, 43 (1982) and adamantylamide L-alanyl-D-isoglutamine (hereinafter AdDP) according to U.S. Pat. AO No 231 221.

These agents have eliminated some adverse properties (eg pyrogenicity) and may be suitable candidates for wider clinical use.

As already discussed in detail (for example, in the description to MS AO No. 231 221), the preparation of these desired substances is often very complicated and expensive. Therefore, in view of the complexity of synthetic work with glycopeptides containing a carbohydrate component in addition to the peptide component, it is always necessary to consider the relative advantages of the biological properties of the substances as opposed to the disadvantageous chemical properties that may later translate into cost.

This could be the case, for example, with known analogs containing a disaccharide component in the molecule (according to PL Durette, EP Meitzner and TS Shen, Carbohydr. Res. 77 (1979)) which, although having an adjuvant effect higher than MDP, are quite difficult accessible.

The MAdDP glycopeptide of the present invention is to a large extent a compromise solution which combines advantageous biological properties with still acceptable complexity of synthesis. A more recent structure-dependent analysis of properties can be found in the aforementioned literature ³ .

The MAdDP glycopeptide of the present invention can be prepared by methods conventional in peptide chemistry by condensing AdDP (prepared according to US AO 231 221) with protected muramic acid.

Protected groups are removed from the protected glycopeptide by, for example, sodium in liquid ammonia or hydrogenation.

Examples

1-Benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy-3-O- (1- (R) -carboxyethyl) -alpha-D-glucopyranoside

1.4 g (3 mmol) of 1-O-benzyl-4,6-O-benzylidene-N-acetylamuramic acid was dissolved in 20 ml of dimethylformamide.

The solution was cooled to -35 ° C and condensed with the pivaloyl chloride procedure. 0.24 (3 mmol) of pyridine, 0.42 ml (3 mmol) of triethylamine and 0.37 ml (3 mmol) of oyl chloride were used. After 10 minutes, the reaction mixture was cooled to -40 ° C and a solution of L-alanyl-D-isoglutamine adamantylamide (prepared according to US AO 231 221) in 10 ml of dimethylformamide was added. The mixture was left overnight at 0 ° C. The product was isolated by precipitation with water.

1.9 g (79%) of 1-benzyl-2-acetoamido-4,6-O-benzylidene-2-deoxy-3-O- [1- (R) -carboxyethyl] -alpha-D-glucopyranoside adamantylamide was obtained.

After purification on a silica gel column (30x5 cm) in ethyl acetate-methanol (9.1), 1.1 g (46%) of a homogeneous material was obtained by thin layer chromatography on silica gel in ethyl acetate-methanol (9.1).

M.p. Mp 224-228 ° C.

Amino Acid Analysis: Glu 1.02, Ala 0.97.

N-Acetylmuramyl-L-alanyl-D-isoglutamine Adamantylamide

0.4 g of the protected peptide prepared by the above procedure was deprotected with a solution of sodium in liquid ammonia. After evaporation of the reaction mixture under reduced pressure, the residue was dissolved in 100 ml of acetic acid acidified water and the solution was chromatographed on a sulfonate cation exchange column in an H ⁺ cycle (20x2 cm), eluting with water. Purification was monitored by HPLC. Stationary reverse phase of ODS type, mobile phase 50% methanol, 50% 0.1% aqueous trifluoroacetic acid. The product was isolated by lyophilization.

137 mg (44%) of N-acetylmuramyl-L-alanyl-D-isoglutamine adamantylamide were obtained.

The substance was homogeneous by thin-layer chromatography in n-butanol, ammonia, water (4.1, 1) systems ( _Rp 0.35); sec. butanol, ammonia, water (85,7,5,7,5) (R _p 0.45) and chloroform, methanol, acetic acid, water (45,30,6,9) (R_ 0,75).

Γ

The structure of the compound of the invention was confirmed by amino acid composition analysis and 1 H-NMR spectroscopy.

Claims (1)

SUBJECT OF THE INVENTION N-Acetylmuramyl-L-alanyl-D-isoglutamine Adamantylamide of Formula HIM CH, OH / in H 3 OH NH Ac CH ₃ - CH - CO - NH - CH - CO - NH - CH - CO - NH ₃ ch ₃ (ch ₂ ) ₂ CO NH Ad where Ad represents the remainder of the adamantane bound in position 1 and Ac is acetic acid residue.