HUT64695A - Method for production of preparations containing nicotinyl-alanine for the treatment of central nervous system (cns) diseares - Google Patents

Abstract

Nicotinylalanine or physiologically equivalent derivatives thereof are used for the preparation of a medicament for the treatment of conditions characterised by an abnormal increase in the metabolism of tryptophan and abnormal excitatory neurotransmission.

Description

The present invention relates to the use of nicotinyl allanine as a pharmaceutical composition for the treatment of diseases of the central nervous system.

The formula of nicotinylalanine is that of formula (I). The compound contains an asymmetric carbon atom such as an enantiomer (R or S) or a racemic form. The invention also relates to the use of enantiomers or racemic mixtures.

Individual enantiomers of nicotylalanine may be prepared from the racemic compound by known optical resolution techniques. The compounds may also be prepared by the stereoselective synthesis described below.

Nitoctinylalanine has been known for about 30 years and is known in the art to be a metabolite of the tryptophan and an intermediate metabolite of the so-called kynurenine pathway, the sequential metabolic pathway of the pathway to open the indole ring of tryptophan to form NAD.

However, it has subsequently been found that this theory cannot be experimentally verified and that nicotylalanine is not a metabolite of tryptophan, although it interferes with its metastatic function in at least two enzyme systems, kynureninase and kynurenine hydroxylase [Decker et al., J. Bioi. Chem. 1963; 238; 1049-1053].

These enzymes catalyze the metabolism of quinurenin first to 3-hydroxy-kynurenine and then to 3-hydroxy-anthranilic acid, the direct precursor of quinolinic acid.

Recently, it has been found that 3-hydroxy-kynurenine can be toxic to nerve cells in the body and that 3-hydroxy-kynurenine may be toxic to nerve cells in the body and Quinolinic acid is a toxin capable of stimulating neurons and likely to kill them, both in vitro and in in vivo formulations (Stone and Connick; Neuroscience, 15, 597-618 (1985)). .

The mechanism responsible for the killing of these nerve cells is that resulting in competitive inhibition between kynurenine hydroxylase and kynureninase. During the metabolic conversion of tryptophan to NAD, kynuric acid may also be produced by a simple enzymatic process in which trans-aminase is involved.

Said acid is capable of inhibiting the toxic stimulatory effect of quinolinic acid by antagonizing the action of the latter on NMDA receptors by modifying the mechanism which has now been clearly elucidated (Moroni et al., Eur. J. Pharmac. 1989; 163; 123-126).

The metabolic conversion of tryptophan and the so-called kynurenine pathway are shown in Scheme 1.

However, based on biochemical knowledge to date, nicotinylalanine was not expected to have any therapeutic effect; although the compound has been known for decades, its pharmacological activity has never been tested in practice.

It has been found that nicotylalanine prevents the accumulation of toxic tryptophan metabolites and increases the synthesis of sedative metabolites as shown in the following pharmacological experiments.

In the biological fluids of rats and mice treated with lipopolysaccharides extracted from the bacterium,

and in the brain, quinolinic acid and kynurenic acid concentrations were three times higher than in control animals after approximately 70 hours. This is due to the induction of the indole amino-2,3-dioxygenase enzyme, which increases the production of substances derived from the kynurenine pathway. When nicotinylalanine (50-1000 mg / kg i.p.) is added to bacterial lipopolysaccharides, the increase in the concentration of quinolinic acid is significantly reduced and the concentration of kynurenic acid is significantly increased. In other words, nicotinylalanine promotes the formation of metabolic products of tryptophan that are antagonistic to the stimulating receptors of amino acids and reduces the production of toxic stimulating agents. Similar results are obtained when nicotinylalanine is administered in an appropriate amount with tryptophan.

In DBA / 2 mice, concomitant administration of tryptophan (200 mg / kg i.p.) and nicotinylalanine (200 mg / kg i.p.) results in a sedative effect on the animals and protection of the animals from audiogenic cramps. This means that the accumulation of kynurenic acid in the brain (as we have shown) is related to the behavior of the animals.

The applicability of nicotinylalanine to various epileptic seizures is tested against leptazole (pentylene tetrazole; metrazole) and electroshock-induced seizures.

10 mice, each weighing 25-35 g, were given nicotinylalanine followed by 1 hour later with leptazole (85 mg / kg subcutaneously). At an intraperitoneal dose of 370 mg / kg, nicotinyl5

-alanine provides protection against epileptic seizures caused by leptazol and increases the latency of seizures from 270 to 588 seconds. Leptazol assay is widely used to test drugs that are effective against petit epilepsy today.

In another experiment, 10 mice were examined to determine the threshold of electrical current required to induce convulsions from the ear electrodes. Animals were treated with 370 mg / kg of nicotinylalanine intraperitoneally and 40% of the animals were protected against seizures. This test is often used as an indicator of drugs that are effective against grand today's epilepsy.

At an effective dose of 370 mg / kg, experimental animals exhibit sedation but minimal ataxia and other movement disorders.

When nicotinylalanine (200-350 mg / kg i.p.) is administered to Swiss mice, the kinurenin i.c.v. dose-induced epileptic seizures can be completely prevented; administration of nicotinyl (150-400 mg / kg) to ethanol-dependent C57 mice prevents the resolution of the onset of epileptic seizures, tremors, and changes in body temperature (Ritzman and Tabakoff 193, 158-170, 1976).

In hippocampal slices of rats incubated in the presence of kynurenine, nicotinyl alanine (0.1-100 μπιόΐ) increases the neosynthesis of kynurenic acid. It is also observed that nicotinylalanine releases radioactive glycine from the binding sides of cortical membranes.

It is clear from the foregoing that nicotinylalanine is useful in the treatment of diseases of the central nervous system resulting from an abnormal increase in tryptophan metastasis.

Examples of diseases in which there is an imbalance between toxic stimulant substances and their modulators or antagonists include:

Bacterial and viral diseases (including

AIDS). In these cases, activation of the immune system causes an increase in the tryptophan metabolic pathway of kynurenine, which results in the accumulation of quinolinic acid in the brain [Heyes et al., Annals Neurol. 1989; Heyes et al., 1988; 51; 1946-1948);

2) Cancerous diseases, particularly diseases affecting the immune system (lymphoid disease);

3) treatment with interferons and / or interleukins (Brown et al., Cancer Res., 1989; 49; 4941-4945);

4) Numerous seizures (Lapin et al., Epilepsy, 1981; 22; 257-265), including grand mal and petit mal epilepsy;

5) Huntington's chorea and other degenerative diseases such as Parkinson's disease and dementia in the elderly (Schwarts et al., Life Sciences, 1984; 35; 19-23);

6) Liver Diseases (Moroni et al., J. Neurochem.,

1986; 46; 849-874);

7) Changes in the sleep-wake cycle;

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8) Neuropsychiatric changes due to pellagra or other vitamin deficiency;

9) Chronic alcoholism, in which the level of kynurenic acid in the brain decreases;

10) Situations of psychiatric interest in which tryptophan metastasis is proven to play a role (depression, schizophrenia, etc.);

11) Stroke and other cerebral ischemic complaints such as traumatic head injury;

12) High blood pressure and basic tremor symptoms.

Nicotinylalanine or its physiologically equivalent derivatives (salts, esters, non-toxic amides) are generally employed in an amount of 10 to 100 mg / kg / day to achieve this effect. The exact dosage will depend on many factors, including the circumstances of the patient, the nature and severity of the disease.

The substance may be administered orally, rectally or parenterally in the form of conventional preparations. Such are described, for example, in Remington's Pharmaceutical Sciences Handbook, Mack Publ. Co., N.Y. USA, 17th Edition, 1985.

Nicotinylalanine can be prepared according to J. Biol. Chem. 238, 1049 (1963) and J. Org. Chem. 28, 383 (1953). Although the compound has the natural L steric configuration of the compound, the invention also encompasses the use of the racemic form and the D form of nicotinylalanine. The enantiomers may be prepared according to Schemes 2 and 3, or as exemplified in the Examples.

Example 1

Preparation of S-nicotinylalanine

a) S- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) acetyl chloride [Compound 3]

S- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) -acetic acid [J.M. Scholtz, P.A. Bartlett Synthesis (1989), 542-543] [2 (0.491 g, 1.76 mmol)] was added to a solution of thionyl chloride in toluene (5 mL, 1: 1 v / v) and the the mixture was stirred at room temperature for 4 hours under argon. After evaporation of the solvent, the residue was dried under gentle vacuum to give pure 3 (0.513 g, 98%).

¹ H-NMR (CDCl ₃ ): δ = 3.52 (2H, d, CH ₂ COCl), 4.25 (1H, t, CH); 5.12 (2H, s, CH ₂ Ph); 5.22 and 55.45 (2H, dd, CH ₂ O); 7.30 (5H, s, aromatics);

¹³ C-NMR (CDCl 3): δ = 46.7, 51.6, 68.4, 78.4, 128.3, 128.6, 135.0, 152.5, 170.0, 171.6.

b) S-3- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) acetylpyridine (Compound 4) (PPl 3) ₂ PdCl ₂ (0.051 g, 0.07 mmol) was added. Compound 3 (0.505 g, 1.70 mmol) and trimethyltin pyridine [Y. Yamamoto, A. Yanagi, Chem. Pharm. Bull., 1982, 30 (5), 1731-1737 (0.595 g, 2.50 mmol) in benzene (20 mL) and the resulting reaction mixture was refluxed for 12 hours under argon. After cooling, the catalyst was removed by filtration through Celite, the filtrate was concentrated in vacuo and the residue was taken up in ethyl acetate (25 mL).

The reaction mixture was diluted, washed with saturated sodium bicarbonate solution (5 mL), water (5 mL), brine (5 mL) and dried over anhydrous magnesium sulfate. After evaporation of the solvent, the residue was subjected to flash chromatography eluting with ethyl acetate / hexane (8: 2). This gave pure 4 (0.276 g, 48%).

1 H-NMR (CDCl ₃ ): δ = 3.40 and 3.70 (2H, dd, CH ₂ CO), 4.40 (1H, t, CH), 5.10 (2H, s, CH ₂ Ph) , 5.40 and 5.53 (2H, dd, CH ₂ O), 7.25 (5H, s, Ph); 7.20-7.40 (1H, m, Py), 7.95-8.10 (1H, m, Py); 8.558.70 (1H, m, py); 8.95 (1H, s, Py);

¹³ C NMR (CDCl ₃ ): δ = 36.9, 50.8, 67.7, 78.4, 123.5, 128.2, 128.5, 131.8, 149.5, 152.5 , 154.0, 171.8, 195.3.

c) s-nicotinylalanine (compound of formula 5)

A suspension of compound (4) (0.087 g, 0.24 mmol) in 6N hydrochloric acid (10 mL) was heated at reflux for 8 hours. The resulting solution was washed with ether (5 mL) and the aqueous phase was evaporated. The residue (0.083 g) was passed through a column of ion exchange resin (Dowex 1x8, AcO-) using 0.3 N acetic acid as eluent. This gives pure 5 (0.046 g, 93%).

158-160 ° C (dec.).

1 H-NMR (D ₂ O): δ = 3.45 (2H, dd, J = 5 Hz and 1 Hz, CH ₂ ), 4.02 (1H, t, J = 5 Hz and 1 Hz, CH ); 7.33 (1H, dd, J = 7.4 Hz and 4 Hz, H-5 Py); 8.19 (1H, dt, J = 7.4 Hz and 1.5 Hz, H-4 Py); 8.45 (1H, dd, J = 4 Hz and 1.5 Hz, H-6 Py); 8.78 (1H, d, J = 1.5 Hz, H-2 Py); 13 C NMR (D ₂ O) δ: 39.0, 50.4, 124.6, 131.6, 136.9, 148.5, 143.3, 173.1, 198.6; [α] _D ²⁵ = + 18.3 (c = 0.6, H ₂ O).

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Example 2

Preparation of R-nicotinylalanine

a) R- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) acetyl chloride [Compound 8]

R- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) acetic acid (7) (0.900 g, 3.22 mmol) was added to a solution of thionyl chloride in toluene (8 mL, 1: 1 v / v). and the resulting mixture was stirred at room temperature for 4 hours under argon. After evaporation of the solvent, the residue was dried under gentle vacuum to give pure compound (8) (0.945 g, 98%).

b) R-3- (3-Benzyloxycarbonyl-5-oxo-4-oxazolidinyl) acetylpyridine (Compound 9) (PPh ₃ ) ₂ PdCl ₂ (0.100 g, 0.13 mmol) is added to a solution of compound (8) (0.900 g, 3.03 mmol) and trimethyltin pyridine (1.10 g, 4.50 mmol) in benzene (30 mL) and the resulting reaction mixture is stirred under argon for 12 hours. reflux. After cooling, the catalyst was removed by filtration through Celite, and the filtrate was concentrated in vacuo, diluted with ethyl acetate (35 mL), washed with saturated sodium bicarbonate solution (10 mL), water (10 mL), brine (10 mL). dried over anhydrous magnesium sulfate. After evaporation of the solvent, the residue was subjected to flash chromatography using ethyl acetate / hexane (8: 2) as eluent. This gave pure 9 (0.279 g, 26%).

c) R-Nicotinylalanine (compound of formula 10)

A suspension of compound (9) (0.270 g, 0.74 mmol) in 6N hydrochloric acid (25 mL) was heated at reflux for 8 hours. The resulting solution was washed with ether (8 mL) and the aqueous phase was evaporated. The residue (250 g) was passed through a column of ion exchange resin (Dowex 1x8, AcO ') using 0.3 N acetic acid as eluent. This gave pure 10 (0.040 g, 31%). Mp 153 ° C. [α] _D 25 = -21 (c = 1, H ₂ O).

Claims (4)

Use of nicotinyl alanine and its physiologically acceptable derivatives as pharmaceutical compositions. 2. Pharmaceutical compositions containing nicotinylalanine or a physiologically equivalent derivative thereof as an active ingredient. Pharmaceutical compositions according to claim 2, wherein the nicotinylalanine L is in a sterile configuration. Use of nicotinylalanine for the manufacture of a medicament for the treatment of diseases which result from an abnormal increase in tryptophan metabolism or from abnormally high neurotransmission.