LT3976B - Novel 3,3'-dithiobis(propionic acids) and esthers thereof - Google Patents

Abstract

This invention describes a new 3.3<1>-ditiobis (propane acid) and their esters, that have immune system modifying effects and the methods of their production, pharmaceutical mixtures, that contain them and their use in medicine.

Description

The present invention relates to novel cystine derivatives having immunomodulatory activity, processes for their preparation, pharmaceutical preparations containing them and their use.

It is an object of the present invention to provide a cystine derivative having immunomodulatory properties. Such a substance is useful in the treatment of various diseases.

N-Acetyl-L-Cysteine is a compound widely used in the treatment of chronic obstructive airway disease / chronic bronchitis (see Multicentre Study Group for further information. Long-term weather acetylcysteine in chronic bronchitis. A double-blind controlled study. A double-blind controlled study. Study J. Eur. Respir. Dis. 1980, 61 (suppl. 111), 93-108; Boman, G., Backer, U., Larsson, S., Melander, B., and Wahlander, L. Weather in acetylcysteine. the rate of exacerbation of chronic bronchitis reported by the Swedish Society for Pulmonary Disease Eur J Respiratory Dis 64, 405-415 (1983) and the British Thoracic Society Research Committee Weather N-acetylcysteine and exacerbation rates in patients with chronic bronchitis and severe airway obstruction. Thorax 1985, 40, 832-835). The mechanism of action of the compound is not disclosed; its action is characterized by mucolytic properties (for further information see Multicentre Study Group. Long-term Weather in Acetylcysteine in Chronic Bronchitis. A Double-blind Controlled Study. Eur. J. Respir. Dis. 1980, 61 (Suppl. , 111), 93-108; Boman, G., Backer, U., Larsson, S., Melander, B., and Wahlander, L. Weather of acetylcysteine exacerbation rate in chronic bronchitis. for Pulmonary Disease, Eur J. Respir Dis.

1983, 64, 405-415; and the British Thoracic Society Research Committee. Weather N-acetylcysteine and exacerbation rates in patients with chronic bronchitis and severe airway obstruction. Thorax 1985, 40, 832-835), antioxidant properties (see Aruoma, O.I., Halliwell, B., Hoey, B.M., and Butter, J. Free Radical Biol. Med. 1989, 6, 593-597), and also immunomodulatory properties. (see Bergstrand, H., Bjomson, A., Eklund, A., Hembrand, R., Eklund, A., Larsson, K., Linden, M., and Nilsson, A. Stimulus-induced superoxide radical generation in vitro by human alveolar macrophages from smokers: Modulation by N-acetylcysteine treatment in vivo (J. Free Radicals Biol. & Med. 2,1986, 119-127).

N, N '- diacetyl - L cystine is also known. This compound has previously been described briefly in both patent and scientific literature (US 4,827,016; EP 300100; US 4,742,439; US 4708965; DE 2326444; Wilson, ID, and Nicholson). , UK Analysis of Thiols and Disulfides in Sulfur Containing Drugs and Related Organic Compounds Chemistry, Biochemistry and Toxicology (ed. LA Damani) Vol 2A Analytical, Biochemical and Toxicological Aspects of Sulfur Xenobiochemistry Ellis Horwood Series in Biochemical Pharmacology (Halstred Press: a division of John Willey & Sons) Chichester 1989, p. 45; and Sjodin,

K., Nilsson, E., Hallberg, A., and Tunek, A. Metabolism of N-acetyl-L-cysteine. Some structural reevaluations for deacetylation and conseuences for weather bioavailability. Biochem. Pharmacol. 1989, 38, 3981-3985). U.S. Patent No. 4,827,016 states that the compound is effective in treating local skin inflammation caused by leukotrienes. However, nothing has been reported or generally known about its pharmacological and / or therapeutic properties related to the immune system and inflammatory lung diseases such as chronic bronchitis.

Simple disulfides having immunostimulatory properties, such as hydroxyethyldisulfide, have previously been described (HEDS, see St. Georgijev, V. New synthetic immunomodulating agents. Trends in Pharmacological Science 1988, 446-51).

Disclosure of the Invention

It has been found that the compound of the present invention has the general formula

COOR ³

I

S - CH ₂ - CH - NH - R

I

S - CH ₂ - CH - NH - CO - R ²

I

COOR ³ I wherein R is hydrogen or a group -CO-R 4 wherein R 4 is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n- nonyl, n-decyl, n-undecyl, iso-propyl, 1-methylpropyl, tert. butyl, 3-methylbutyl 2 or 2-methylbutyl, R is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n -undecyl, iso-propyl, 3

1-methylpropyl, tert. butyl, 3-methylbutyl or 2-methylbutyl, and R is hydrogen, methyl, ethyl, propyl, isopropyl, butyl or isobutyl, provided

3 provided that R and R are not simultaneously methyl and further provided that when R is

2 hydrogen, R and R are not simultaneously n-propyl or n-heptyl, or a physiologically acceptable salt thereof and / or a stereochemically isomeric form thereof, are immunomodulatory, in particular immunostimulatory.

Therefore, the compounds of the invention may be used to treat and suspect diseases with an immune defect and / or defective immune system.

Examples of such diseases are chronic bronchitis and other inflammatory diseases of the airways such as asthma and rhinitis, as well as certain forms of autoimmune diseases such as diabetes and rheumatoid arthritis and / or various malignancies. These compounds can be used to treat HIV infection and AIDS. These compounds can also be used to treat atherosclerosis.

Preferred compounds of formula I above are those wherein R is hydrogen or the group -CO-R 4 wherein R 4 is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, isopropyl, 1-methylpropyl, tert. butyl, 3-methylbutyl or 2-methylbutyl, R is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n- undecyl, iso-propyl, 1-methylpropyl, tert. butyl, 3

3-methylbutyl or 2-methylbutyl, and R is hydrogen, methyl, ethyl, propyl, 1 2 isopropyl, butyl or isobutyl, provided that R and R are not simultaneously 3 to 1-2 methyl, and further provided that that when R is hydrogen, R and R are not simultaneously n-propyl or n-heptyl.

According to formula I, when R is hydrogen, those compounds are preferred wherein R and R, respectively, are simultaneously isopropyl or tert. butyl.

According to the above formula I, when RΨ is hydrogen, it is preferable to 1 2 for those compounds wherein R and R have from 3 to 7 carbon atoms.

Particular preference is given to the compounds of formula I above when

12 3 12

RΨ for hydrogen wherein R and R are isopropyl and R is methyl; R and R

12 3 is n-pentyl and R is methyl; R and R are n-heptyl and R is methyl or ethyl.

Accordingly, L-isomeric stereoisomeric forms of the compounds, i.e., compounds derived from L-cystine, are preferred in the present invention.

The physiologically acceptable salt of the compounds of formula I wherein R is hydrogen is a sodium, ammonium, calcium or magnesium salt together with their non-toxic acid addition salts. Also included are salts derived from arginine, lysine, histidine, ethanolamine, diethanolamine, ethylenediamine and choline, or other suitable organic amines.

The physiologically acceptable salt of the compounds of formula I wherein R Φ for hydrogen and R = hydrogen is hydrochloride, hydrobromide, hydrosulfate, oxalate, tartrate, and the like. Salts can also be solvates, i.e. in the form of hydrates.

Pharmaceutical Formulations

The active ingredients described may be in a variety of dosage forms, e.g. tablets, cachets, gelatin capsules, solutions and aerosols.

For the production of tablets, cachets and gelatin capsules, the active ingredient may be combined with pharmaceutically acceptable substances, pnz. lactose, starch, dicalcium phosphate, microcrystalline cellulose, polyvinylpyrrolidone, gelatin, cellulose derivatives, colloidal silica, talc and stearic acid or its salts.

Suitable media for oral solutions are water, sucrose, glucose, sorbitol, fructose and xylitol.

The dosage forms include, in addition to the aforesaid media, a preservative, a stabilizer, a viscosity regulator, an emulsifier, a sweetener, a coloring agent, a flavoring agent, a tonicity agent, a buffer or an antioxidant. They may also contain other therapeutically valuable substances.

The ways of receiving

According to the invention, the compounds can be obtained in any of the following ways:

A. Oxidation of N-Acylcysteine Derivative Formula

COOR ³

I

HS - CH ₂ - CH - NH - CO - R ¹ ,

3 wherein R ¹ and R ^J are as defined above or when R ^J is hydrogen, the salt of the compound is selected until the compound of the formula is formed

COOR ³

I

S - CH ₂ - CH - NH - CO - R ¹ I

S - CH ₂ - CH - NH - CO - R ¹ I

COOR ³

How the oxidant can be used:

Ί

Hydrogen peroxide, alkaline air, alkylhydroperoxides, acid, halogen, trivalent or divalent nitrogen oxides, oxidizing metals such as thallium (III), trialkylsulfonium salts, or selenium or tellurium oxides. Oxidation can also be performed electrochemically.

Halogens, for example, include chlorine, bromine or iodine. Salts in Method A and the following may be used in the form of sodium, potassium, calcium, ammonium and the like. salts.

B. Reaction in the presence of a suitable base of a compound of the formula

COOR ³

I

S - CH ₂ - CH - NH ₂

I s - ch ₂ - ch - nh ₂

I

COOR ³ or a salt thereof, wherein R is as described above, with a compound of the formula

R ¹ is COX as described above and COX is a reactive group capable of reacting with an amino group to form an amide group to form a compound of formula

COOR ³

I

S - CH ₂ - CH - NH - CO - R ¹ I

S - CH ₂ - CH - NH - CO - R ¹ I

COOR ³ δ

The acylating agent R? COX may be, for example, an acid halide, anhydride, amide, or an activated acid or ester.

The salt in Method B and any of the following processes in which the salt may be used may be hydrochloride, hydrobromide, hydrosulfate, oxalate, tartrate and the like. The salt in Method B may also be any of the alkali salts mentioned in Method A.

C. Reaction Formula of 2- (N-acylamino) -3-halopropionic acid derivative

COOR ³¹

I

Y - CH ₂ - CH - NH - CO - R ¹ ,

Wherein R ¹ is as defined above, R ^J is R ^J as described above, or an acidic or basic labile organic group and Y is a halogen atom, with a sulfur or disulfide dianion in the presence of a base, resulting in the compounds of formula

COOR ³¹

CH ₂ - -CH-NH - - CO CH ₂ - - CH - NH - - CO | COOR ³¹

followed by formation of a compound wherein R ^J = H, removing the 3I protecting group R.

U

The protecting group, R ^J , may be an acidic or basic labile organic group such as an alkyl, benzyl, aryl, vinyl or allyl group.

D. Oxidation of N-Acylcysteine Derivative Mixture Formulas

COOR ³

I

HS - CH ₂ - CH - NH - CO - R ¹ and

COOR ³

I

HS - CH ₂ - CH - NH - CO - R ²

Wherein R, R and R are as described above or when R is hydrogen, it is the possible alkaline salt thereof which results in the following formula:

COOR ³

I

S - CH ₂ - CH - NH - CO - R ¹ I

S - CH ₂ - CH - NH - CO - R ² I

Formation of COOR ³ .

The oxidizing agents described in Method A may be used.

E. Oxidation of formulas of a mixture of cysteine or a cysteine ester with an N-acylcysteine derivative

HS

COOR ³

I

- CH ₂ - CH - NH ₂ or their salts and

COOR ³

I

HS - CH ₂ - CH - NH - CO - R ¹

3 or their alkaline salts, wherein R and R are as described above, to give a compound of the formula

S

I

S

COOR ³ | ch ₂ - CH - NH ₂ ch ₂ - CH - NH - CO I COOR ³

Formation of R ¹ or a salt thereof.

The oxidizing agents used in Method A may be used for oxidation.

F. Reaction in the presence of a suitable base and in the presence of an excess of cystine or cystine diester formula

COOR ³

S - CH ₂ - CH - NH ₂ I

S - CH ₂ - CH - NH ₂ I

COOR ³ or a salt thereof, wherein R is as described above, with a compound of the formula wherein R

R ² is COX, and COX is as described above for the compound of formula

COOR ³

S - CH ₂ - CH - NH ₂

S - CH ₂ - CH - NH - CO - R ² I

Formation of COOR ³ .

G. Reaction in the presence of a suitable base of a compound of the formula

COOR ³

I

CH ₂ - CH - NH - CO

CH ₂ - CH - NH ₂ COOR ³

R ¹ · 3 or a salt thereof, wherein R and R are as described above, with a compound of the formula

R ² is COX wherein R and COX are as defined above, resulting in the compound of formula

COOR ³

I

S - CH ₂ - CH - NH - CO - R ¹ I

S - CH ₂ - CH - NH - CO - R ² I

Formation of COOR ³ .

H. Reaction N-acylcysteine derivative formula

COOR ³

I

HS-CH ₂ -CH-NH-CO-R ¹ or salts thereof with an activating agent such as diethyl azodicarboxylate to form an adduct, e.g.

COOC ₂ H ₅ COOR ³

I I

N - S - CH ₂ - CH - NH - CO - R ¹ I

NH

I

COOC ₂ H _{5 by} further reaction with a second, different or identical N-acylcysteine or N-acylcysteine ester to form a compound of formula

COOR ³

I

S - CH ₂ - CH - NH - CO - R ¹ I

S - CH ₂ - CH - NH - CO - R ² I

COOR ³

I. Reaction of compound of formula

COOR ³¹

and R ¹ and R are as described above, with a compound of the formula

COOR ³¹ I

H - S - CH ₂ - CH - NH - CO - R ² wherein R 1 and R 6 are as described above, leading to the COOR ^{3 of the} compound of formula

ch ₂ - -CH-NH - - co CH ₂ - - CH - NH - - co COOR ³¹

followed by removal of the protecting group R ³ if R = H is desired.

J. Esterification of a compound of formula s

I s

cox ¹ I ch ₂ - I - CH - NH - - R ch ₂ - - CH - I NH - - CO I cox ¹

Wherein R and R are as described above and X is OH or halogen, with a compound of formula

R ^3E - OH,

3E wherein R is methyl, ethyl, propyl, isopropyl, butyl or isobutyl, resulting in the compound of formula

S

I

S

coor ^3E i CH ₂ - I -CH-NH - - R ch ₂ - - CH - NH - J -CO-R ² 1 coor ^3E

formation.

K. Alkylation of the compound of formula

coo ^(_) 1 S - CH ₂ 1 - CH - NH s - ch ₂ - CH - NH COO ^(_)

R

CO - R ² where R and R ² are as defined above, with a compound of formula r ^3E - z,

3E wherein R is as described above and Z is halogen, alkylsulfate, tosylate or another nucleophile that results in the formation of a compound of formula C00R ^3E I

S - CH ₂ - CH - NH - RI

Formation of S - CH ₂ - CH - NH - CO - R ² I coor ^3E .

L. Reaction of Carboxyl-Protected Cystine Derivative in Suitable Base Formulas

COOR ^3P

I

S - CH ₂ - CH - NH ₂ I

S - CH ₂ - CH - NH ₂ I

COOR ^3P

Reaction of 3P or its hydrochloride salt, where R is an acidic or basic labile organic group, with a compound of the formula

R ¹ is COX wherein R ¹ is as described above and COX is a reactive group capable of reacting with an amino group to form an amide group followed by deprotection of 3P

R, to form a compound of formula

COOH

I s - ch ₂ - ch - nh - co - r ¹ s - ch ₂ - ch - nh - co - r ¹ I

COOH

3P

The acylating agent R COX is as described in B. The protecting group R 1 may be an acidic or basic labile organic group such as an alkyl, benzyl, aryl, vinyl or allyl group.

M. Reaction of a carboxyl-protected cystine derivative in an alkaline environment

coor ^3P ch ₂ - 1 CH - NH ch ₂ - CH - NH _: 1 COOR 3P

3P or its hydrochloride salts, wherein R is as described above, with a compound of the formula

R ^{2 is} -COXi

3P wherein R and COX are as described above and after removal of the protecting group R resulting in the compound of formula

COOH

I

S - CH ₂ - CH - NH ₂

I

S - CH ₂ - CH - NH - CO - R ² I

COOH formation.

N. Reaction of carboxyl-protected cystine derivative of formula

COOR ^3P

I

S - CH ₂ - CH - NH ₂ I

S - CH ₂ - CH - NH - CO - R ² I

COOR ^3P

3P or its hydrochloride salt, wherein R and R are as described above, with a compound of formula

R ¹ for COX,

3P wherein R and COX are as described above and after removal of the protecting group R resulting in the compound of formula

COOH ch ₂ 1 - CH - NH - CO ch ₂ - CH - NH - CO

COOH formation.

O. Cysteine derivative and cysteine derivative of the formula COOH 1

S - CH ₂ - CH - NH - CO - R ¹

S - CH ₂ - CH - NH - CO - R ¹ I

COOH

COOH

I

HS - CH ₂ - CH - NH - CO - R ²

2 or a mixture of their alkaline salts, wherein R and R are as described above, in equilibrium in an alcoholic or aqueous solution, resulting in a compound of formula

S

I

S

COOH CH ₂ 1 - CH - NH - CO - R ¹ ch ₂ - CH | NH - CO - R ² COOH

formation.

In all the A-0 routes, the resulting compound can be converted into a physiologically acceptable salt.

Working examples

An example. (R, R) -N, N '-dipeptanoyl -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester: suspension of L-cystine dimethyl ester dihydrochloride (1.0 g, 3 mmol) in tetrahydrofuran -THF- (20 ml) (white liquid) ) was stirred and cooled to 0 ° C. 4 eq. (2.0 mL) of N-ethyldiisopropylamine and 2.2 eq. (0.8 mL, 6.6 mmol) pentanoyl chloride. The mixture was stirred in an ice bath for 4 hours and the white precipitate of N-ethyldiisopropylammonium chloride was removed by filtration. The solvent was removed by evaporation and the residue was redissolved in dichloromethane. After washing with water, the organic phase was dried over sodium sulfate. Filtration and evaporation of the solvent afforded the desired crude compound which was recrystallized from ethyl acetate.

Physical data:

1 H-NMR (300 MHz, CDCl 3: d 6.42 (2H, d, NH), 4.88 (2H, dt, NCH), 3.77 (6H, s, OCH ₃ ), 3.21 (4H, m, SCH ₂ ), 2.27 ( 4H, t, COCH ₂ ), 1.63 (4H, m, CH ₃ CH ₂ CH ₂ ), 1.36 (4H, m, CH ₃ CH ₂ ), 0.92 (6H, t, CH 2.). [A] ² ^ -146 ° (C = 0, 490, MeOH) Melting point = 110-112 ° C Analysis calculated:% C ₁₈ H ₃₂ O ₆ N ₂ S ₂ : C 49.52; H 7.39; N 6.42; S, 14.69 Found: C, 49.40; H, 7.40; N, 6.40; S, 14.70.

An example. (R, R) -N, N '-dipropionyl -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester: The compound was synthesized according to the procedure described in Example 1 using propionyl chloride as the acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl 3: d 6.44 (2H, d, NH), 4.89 (2H, dt, NCH), 3.78 (6H, s, OCH ₃ ), 3.22 (4H, m, SCHp, 2.31 (4H, q, CH ₃ CH ₂ CH ₂ ), 1.18 (6H, t, CHp.

An example. (R, R) -N, N '-dipentanoyl -3,3'-dithiobis (2-aminopropanoic acid) diethyl ester: The compound was synthesized according to the procedure described in Example 1 starting with L-cystine diethyl ester dihydrochloride and using pentanoyl chloride as an acylating agent.

Physical data:

¹ H-NMR (300 MHz, CDCl 3): δ 6.43 (2H, d, NH), 4.85 (2H, dt, NCH), 4.23 (4H, m, OCH ₂ ), 3.22 (4H, m, SCHp, 2.27 (4H, t, COCH ₂ ), 1.63 (6H, m, CH ₃ CH ₂ CH ₂ ), 1.37 (4H, m, CH ₂ CH ₂ , 1.30 (6H, t, OCH 2 H), 0.92 (6H, t, [Α] D 25 = -30 ° (C = 0.514, MeOH) Melting point = 109 ° C Analysis calculated:%:

^C 20 ^H 36 ^N 2 ° 6 ^{S 2: C} '· ^{51 7θ; H} '^{7-81; N} ' ⁶ · ⁰³ ' ^{S '13} · ⁸⁰ · ^Found ' ^{%: C} '^{5L85; H} , 7.75; N, 6.10; S, 13.60.

An example. (R, R) -N, N '-dipropionyl -3,3' -dithiobis (2-aminopropanoic acid) diethyl ester: The compound was synthesized according to the procedure described in Example 1 starting with L-cystine diethyl ester dihydrochloride and using propionyl chloride as the acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl ₃ ): δ 6.44 (2H, d, NH), 4.86 (2H, dt, NCH), 4.23 (4H, m, OCH ₂ ), 3.22 (4H, m, SCH ₂ ), 2.31 (4H, q, COHCH ₂ ), 1.30 (6H, t, CO ₂ CH ₂ CH ₃ ), 1.18 (6H, t, CH ₃ ).

An example. (R, R) -N, N '-Dihexanoyl -3,3'-dithiobis (2-aminopropanoic acid) diethyl ester: The compound was synthesized according to the procedure described in Example 1 starting with L-cystine diethyl ester dihydrochloride and using hexanoyl chloride as an acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl 3): δ 6.44 (2H, d, NH), 4.86 (2H, dt, NCH), 4.23 (4H, m, OCH ₂ ), 3.21 (4H, m, SCHp, 2.26 ( 4H, t, COCH ₂ ), 1.65 (4H, m, COCH ₂ CH ₂ ), 1.31 (4H, m, CH ₃ CH ₂ CH ₂ ), 1.31 (4H, m, CH ₃ CH ₂ ), 1.31 (6H, t, OCH ₂ CH ₃ ), 0.90 (6H, t, CH ₃ ).

An example. (R, R) -N, N '-di (2-methylpropionyl) -3,3'-dithiobis (2-aminopropanoic acid) diethyl ester: The compound was synthesized according to the method described in Example 1 starting with L-cystine diethyl ester dihydrochloride and using 2-methylpropanoyl chloride as acylating agent. material.

Physical data:

1 H-NMR (300 MHz, CDCl ₃ ): δ 6.42 (2H, d, NH), 4.84 (2H, dt, NCH), 4.23 (4H, m, OCH ₂ ), 3.22 (4H, m, SO 2, 2.46 ( 2H, m, (CH 2 CH), 1.30 (6H, t, OCH ₂ CH ₃ ), 1.18 (12H, d, (CH ₃ ) ₂ Q. [α] ²⁰ D = -127 ° (C = 0.512, MeOH) Melting point = 140-141 ° C Analysis: Calculated:% ^C 18 ^H 32 ° 6 ^N 2 ^S 7 ^: C 49.52; H 7.39; N 6.42; S 14.69 Found:% C, 49.15; H, 7.40; N, 6.30; S, 14.75.

An example. (R, R) -N, N '-di (1-Oxo-dodecanyl) -3,3' -dithiobis (2-aminopropanoic acid) dimethyl ester:

The compound was synthesized according to the method described in Example 1 starting with (R, R) -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester dihydrochloride and using dodecanonic acid chloride as the acylating agent.

Overall yield: 40%

Physical data:

M.p. = 98-99 ° C. [.alpha.] @ ²⁵ = -93 DEG (C = 0.531, MeOH / d6 H-NMR (300 MHz, CDCL): d 6.42 (2H, d, NH), 4.88 (2H, dt, NCH), 3.77 (6H, s, OCHp 3.21 (4H, m, _SCH?), 2.26 (4H, t, OCH _2), 1.64 (4H, m, OCH ₂ CH _2), 1.26 (32H, m, (CH _{2) 8),} 0.88 (6H , t, CH ₂ CH ₂ ) Analysis: Calculated:% C, 60.72; H, 9.56;

4.43; S, 10.13.

Found,%: C, 60.4; H, 9.3; N, 4.5; S, 10.1.

An example. (S, S) -N, N '-di (2-methylpropionyl) -3,3' -dithiobis (2-aminopropanoic acid) dimethyl ester:

N, N '-diisobutyryl-D-cystine (1.86 g, 4.9 mmol) was dissolved in 10 mL of methanol containing one drop of hydrochloric acid. Trimethylorthoformate (0.6 ml,

5.5 mmol) and the reaction mixture was stirred at room temperature for 4 days. After evaporation of the solvent, the product was purified on a chromatography column (solvent heptane: ethyl acetate 1: 5).

Physical data:

Melting point = 145.5-147.5 ° C. 1 H-NMR (300 MHz, CDCl ₃ ): δ 6.39 (2H, d, NH), 4.86 (2H, dt, NCH), 3.78 (6H, s, OCH ₂ ), 3.21 (4H, m, SCH ₂ ), 2.47. (2H, h, CH (CH ₃ ) ₂ ), 1.18 (12H, d, CH (CH ₃ ) ₂ ). [Α] ²⁰ D = + 135.2 (C = 0.42, MeOH)

An example. (R, R) -N, N '-Dihexanyl-3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester: The compound was synthesized according to the procedure described in Example 1 using hexanoyl chloride as an acylating agent.

Physical data:

¹ H-NMR (300 MHz, CDCl 3): δ 6.41 (2H, d, NH), 4.88 (2H, dt, NCH), 3.76 (6H, s, OCH ₃ ), 3.21 (4H, m, SCH ₂ ). , 2.26 (4H, t, COCH ₂ ), 1.65 (4H, q, CH ₃ CH ₂ CH ₂ CH ₂ ), 1.33 (4H, m, CH ₃ CH ₂ , 1.33 (4H, m, CH ₃ CH ₂ ), 0.89 (6H, t, CH ₃ ). [Α] ²⁰ J = -135 ^° (C = 0.486, MeOH) m.p. = 90-92 ° C.

Calculated,%: C 2 _Q H ₃₆ N ₂ O ₆ S ₂ : C, 51.70; H, 7.81; N, 6.03; S, 13.80.

Found,%: C, 51.90; H, 7.95; N, 6.10; S, 13.75.

An example. (R, R) -N, N '-di (1-Oxo-octyl) -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester: The compound was synthesized according to the procedure described in Example 1 using octanoyl chloride as the acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCL): d 6.41 (2H, d, NH), 4.88 (2H, dt, NCH), 3.77 (6H, s, OCH ₃ ), 3.21 (4H, m, SCH ₂ ), 2.26. (4H, t, COCHp, 1.65 (4H, t, COCH ₂ CH ₂₎

1.29 (16H, m, CH ₃ (CH ₂ ) 1-4, 0.88 (6H, t, CH ₂ CH ₃ ). [Α] ²⁰ D = -119 ° (C = 0.490, MeOH). M.p. 90-91 [deg.] C. Analysis: Calculated:%: C, 54.80; H, 8.45; N, 53.80; H, 8.52; N, 5.38; S, 12.31. , 5.35; S, 11.80.

An example. (R, R) -N, N '-di (1-Oxo-octyl-3,3'-dithiobis (2-aminopropanoic acid) diethyl ester: The compound was prepared according to the procedure described in Example 1 starting with L-cystine diethyl ester dihydrochloride. octanyl chloride as an acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl 3: d 6.43 (2H, d, NH), 4.85 (2H, dt, NCH), 4.23 (4H, m, CH ₃ CH ₂ O), 3.21 (4H, m, SCH ₂ ). , 2.26 (4H, t, COCH ₂ ), 1.65 (4H, t, COCH ₂ CH ₂ ), 1.30 (6H, m, OCH ₂ CH ₃ ), 1.30 (16H, m, CH ₃ (CH ₂ ) 1-4 ), 0.88 (6H, t, ch ₂ ch ₃ ).

An example. (R, R) -N, N '-di (2,2-Dimethylpropionyl) -3,3' -dithiobis (2-aminopropane fatty acid) diethyl ester: The compound was prepared according to the procedure described in Example 1 starting with L-cystine diethyl ester dihydrochloride.

2,2-dimethylpropanoyl chloride as an acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl 3: d 6.52 (2H, d, NH), 4.80 (2H, dt, NCH), 4.22 (4H, m, CH ₃ CH ₂ O), 3.22 (4H, m, SCH ₂ ). , 1.30 (6H, t, OCH ₂ CH ₃ ), 1.23 (18H, s, C (CH ₃ ) ₃ ).

An example. (R, R) -N, N '-di (2,2-Dimethylpropionyl) -3,3' -dithiobis (2-aminopropanoic acid) dimethyl ester: Compound prepared according to the procedure described in Example 1 using 2,2-dimethylpropanoyl chloride. as an acylating agent.

Physical data:

1 H-NMR (300 MHz, CDCl 3): δ 6.51 (2H, d, NH), 4.83 (2H, dt, NCH), 3.78 (6H, s, OCH ₃ ), 3.21 (4H, m, SCH ₂ ). , 1.23 (18H, s, C (CH ₃ ) ₃ ).

An example. (R, R) -N, N '-di (2-Methylpropionyl) -3,3' - dithiobis (2-aminopropanoic acid) dimethyl ester: Compound prepared according to the procedure described in Example 1 but substituted with pentanoyl chloride with 2-methylpropionyl chloride.

Physical data:

Melting point = 142-5 ° C. d 6.40 (2H, d, NH), 4.86 (2H, dt, NCH), 3.78 (6H, s, OCH

3.21 (4H, m, SCH ₂ ), 2.47 (2H, h, CH (CH ₃ ) ₂ ), 1.18 (12H, d, CH (CH ₃ ) ₂ ).

An example. (R, R) -N-Acetyl-N'-hexanoyl-3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester: (R, R) -3,3'-Dithiobio (2-aminopropanoic acid) dimethyl ester dihydrochloride ( 690 mg, 2 mmol) was mixed with 1.39 mL (10 mmol) of triethylamine in 20 mL of tetrahydrofuran in a 100 mL round bottom flask. The cloudy solution was cooled to 0 ° C in an ice bath, followed by dropwise addition of a solution of acetyl chloride (142ml, 2mmol) and hexanoyl chloride (276ml, 2mmol).

In 3 mL of tetrahydrofuran. The cloudy solution was stirred for 1 h at 0 ° C. 1 g (100%) of a white precipitate of Et ₃ N «HCl was filtered off and the filtrate was evaporated until an indifferent residue was obtained which was partitioned between 10 mL of H ₂ O and 10 mL of CHCl 3. , the aqueous phase was extracted with ₃ x ₁₀ mL CHCl ₃ . After evaporation of the combined CHCL-phases, the crude product (containing the desired material and symmetrical derivatives) was isolated by flash chromatography according to Stille and co-authors (J. Org. Chem. 1978, 43, 2923) on silica gel 60 (E. Merck 5735, 230). -400 mesh ASTM) with EtOAc / heptane / MeOH 6: 3: 1 as eluent. Separation was continued by thin layer chromatography (Silica gel plastic sheets 60 Wf ₂ 54s, E. Merck 16483, eluent EtOAc / Heptane / MeOH 6: 3: 1 and I ₂ as visualizer). The fractions of acceptable purity were combined and evaporated until an indifferent residue was obtained. This residue was dissolved in acetone and the solution was evaporated again until a colorless solid was obtained.

Yield: 184 mg, 25%.

Physical data:

TLC: Rf = 0.30 (EtOAc / heptane / MeOH = 6: 3: 1). 1 H-NMR (300 MHz, CDCl 3): δ 6.53 (1H, d, NH), 6.43 (1H, d, NH), 4.88 (2H, m, NCH), 3.78 (6H, s, CO ₂ CH ₃ ). , 3.22 (4H, m, SCH ₂ ), 2.26 (2H, t, COCH ₂ ), 2.07 (3H, s, COCH ₃ ), 1.65 (2H, m, CH ₂ ), 1.32 (4H, m, CH ₂ ). , 0.90 (3H, t, CH ₃ ). LAPMS (m / z): 431 [MNa] ⁺ , 409 [MH] ⁺ , 311 [MH-C _? H ₁ jO] *

Example: (R, R) -N, N'-Di (2-Methylpropionyl) -3,3'-Dithiobis (2-Aminopropanoic Acid): N-Isobutyryl-L-Cysteine (9.5 g, 50 mmol) dissolved in 50 ml of water. Hydrogen peroxide (30%, 3.1 mL 30mmol) was added and the reaction mixture was stirred at room temperature for 6 hours. Evaporation of the solvent under reduced pressure gave a white, crystallizing oil (9.8 g). After recrystallization from ethyl acetate, the title compound is obtained in the form of a white solid and dried under vacuum.

Yield: 4.8 g (50%) Physical data: m.p. 143-5 ° C; 1 H-NMR (300 MHz, DMSO-d ₆ ): d (2H, b, CO ₂ H), 8.16 (2H, d, NH), 4.47 (2H, m, CHN), 3.15 (2H, dd, CH). ₂ d, J = 14 Hz, 5Hz), 2.92 (2H, dd, CH ₂ S, J = 14 Hz, 9 Hz), 2:43 (2H, h,

D

CH (CH ₃ ) ₂ , J = 7Hz), 1.01 (12H, d, CH ₃ , J = 7Hz). [α] ²⁵ = -1.69.8 ° (MeOH, C = 0.510).

An example. (R, R) N-Acetyl-3,3 '-dithiobis (2-aminopropanoic acid):

L-cysteine (2.42 g, 20 mmol) and N-acetyl-L-cysteine (3.26 g, 20 mmol) were dissolved in 25 mL of water. The pH of this solution according to litmus paper was 2.6. Hydrogen peroxide (30%, 2.3ml, 21mmol) was added and the reaction mixture was left overnight at room temperature. The white precipitate stood at room temperature overnight. The white precipitate was filtered off to give (1.26 g). Comparison of the spectrometric data with an authentic sample showed L-cystine. The yellowish filtrate containing the desired compound and two symmetrical compounds (R, R) - N, N'-diacetylcystine and (very small amount) of cystine - was separated by high performance liquid chromatography (hereinafter ASSC) with a Dynamax C8 column (8 mm). , 60 A, 21.4 x 250 mm) with Dynamax C ^ -protection column (8 mm, 21.4 x 50 mm) and Gilson dual solvent delivery system (305 pump, 100 SC pressure pump acting as solvent delivery pump, pressure pump 5 SC acting as a sample injector, 806 pressure gauge module, 81 IB dynamic mixer, 115 UV detector, 201 fractional collector, 201-202 fractional controller). Solvents used: A = 10 mM HOAc / P ^ O and B = MeOH, A: B in the ratio 95: 5. The solvent current was 10 mL / min and the release was recorded at 230 nm. After determination of both fractions by thin-layer chromatography (hereinafter referred to as PSC) (Merck 16483, silica gel 60WF 254s, eluent n-BuOH / HOAc / FLjO 1: 1: 1, ₁₂ as the developer), the fractions of acceptable purity were collected and evaporated to give an oily residue. . This residue was dissolved in acetone (pa), evaporated to give the desired product as white crystals. Yield: 10%.

Physical data:

TLC: R _f = 0.69 (n-BuOH / HOAc / H ₂ O = Fl.-l ^ H-NMR (300 MHz, D ₂ O) d 4.73 (1H, dd, NCH), 4.18 (1H, dd, NCH), 3.37 (2H, m, SCH ₂ ), 3.10 (2H, m, SCH ₂ ), 2.06 (3H, s, CH ₃ ) TSP-MS (m / z): 283 [MH] ⁺ , 164 [ MH-C ₃ H ₅ NC> ₂ ] ⁺

An example. (R, R) -N-Acetyl-N '- (2-methylpropionyl) 3,3'-dithiobis (2-aminopropanoic acid): N-acetyl-L-cysteine (0.652 g, 4 mmol) and Nisobutyryl-L- A mixture of cysteine (0.764 g, 4 mmol) was stirred in 10 mL of MeOH while adding dropwise hydrogen peroxide (30%, 0.60 mL, 5 mmol). Stirring was continued at room temperature for 3 hours, after which the solvent was removed by rotary evaporation. Addition of 25 ml of acetone and repeated evaporation gave 1.5 g of crude oil which solidified on standing. The desired compound was isolated from this mixture using ASCC as described in Example 17. Yield: 31%.

Physical data:

Elution of DESC with 60% A (isocratic, see Example 17 for solvents) .PSC: R ^. = 0.76 n-BuOH / H ₂ O / HOAc / = 1.H1 ^ H-NMR (300 MHz, D ₂ O). : d 4.73 (2H, m, NCH), 3.33 (2H, m, SCH ₂ ), 3.03 (2H, m, SCH ₂ ), 2.59 (1H, n, H), 2.06 (3H, s, COH ₃ ), 1.13 (3H, d, CHp, 1.11 (3H, s, CHp. Plasma Absorption Mass Spectrometry (PAMS)) (m / z): 376 [MNa] ⁺ , 353 [MH] ⁺ .

An example. (R, R) N- (2-Methylpropionyl) -3,3'-dithiobis (2-aminopropanoic acid) The compound was prepared according to the procedure described in Example 17 starting with L-cysteine and N-isobutyryl-L-cysteine. Yield: 8%.

Physical data: ASCC elution with 55% A (isocratic, see Example 17 for solvents). PSC: R _f = 0.67 n-BuOH / HpHOAc = 1/1/1 1 H-NMR (300 MHz, D ₂ O):

d 4.73 (1H, dd, NCH), 4.14 (1H, dd, NCH), 3.38 (2H, n, SCH ₂ ), 3.08 (2H, m, SCH ₂ ), 2.06 (1H, n, CH), 1.15 ( 3H, d, CH 3, 1.13 (3H, d, CH ₃ ). TSP-MS (m / z): 311 [MH] ^+.

An example. (R, R) - N-Acetyl-N '- (2,2-dimethylpropionyl) -3,3' -dithiobis (2-aminopropanoic acid): The compound was prepared according to the procedure described in Example 18 starting with N-acetyl-L. -cysteine and N-pivalyl-L-cysteine. Yield: 21%. Physical data:

ASCC elution gradient: 50% A / 15 min, 50 30% A / 5 min,

R _f = 0.78 ⁿ BuOH / H ₂ O / HOAc = 1: 1: 1).

¹ H-NMR (300 MHz, D ₂ O): d 4.72 (2H, m, NCH), 3.35 (2H, m, CH ₂ ), 3.04 (2H, m, SCH ₂ ), 2.06 (3H, s, CH ^ 1.21 (9H, s CCH ^.

PAMS (m / z): 411 [MNa] ⁺ , 389 [MNa] ⁺ , 367 [MH] ⁺ , 349 [MH-H ₂ O] ⁺ .

An example. (R, R) -N, N'-Di (2,2-Dimethylpropionyl) -3,3'-dithiobis (2-aminopropanoic acid): The compound was isolated from the crude material obtained in Example 20. Yield: 15%.

Physical data:

ASCC Elution Gradient: 50% A / 15 min, 50 30% A / 15 min, 30%. A / isocratic (for solvents see Ex. 17 PSC: Rf = 0.78 ⁿ BuOH / H ₂ O / HOAc = 1: 1: 1).

₁ H-NMR (300 MHz, D ₂ O): d 4.72 (2H, dd, NCH), 3.37 (2H, dd, SCH ₂ ), 3.06 (2H, dd, SCH ₂ ), 1.21 (18, s, CCH _3). ).

PAMS (m / z): 431 [MNA] ⁺ 409 [MH] ⁺ 391 [MH-H ₂ O] ^+.

An example. (R, R) - N-Acetyl-N'-pentanoyl-3,3-dithiobis (2-aminopropanoic acid):

This compound was prepared according to the procedure described in Example 18 starting with N-acetylcysteine and N-pentanoylcysteine. Yield: 23%.

Physical data:

ASCC Elution Gradient: 50% A / 20 min, 50 25% A / 5 min, 25%

A / isocratic, (see Example 17 for solvents). PSC: R f = 0.84 n-BuOH / F ^ O / HOAc = 1: 1: 1).

₁ H-NMR (300 MHz, D ₂ O): δ 4.70 (2H, m, NCH), 3.30 (2H, n, SCH ₂ ), 2.99 (2H, m, SCH ₂ ), 2.30 (2H, t, COCH 3). 2.04 (3H, s, COCH?, 1.57 (2H, m, CH ₂ ),

1.30 (2H, m, CH ₂ ), 0.86 (3H, t, CH ₃ ).

PAMS (m / z): 389 [MNA] ⁺ 367 [MH] ⁺ 349 [MH-H ₂ O] ^+.

An example. (R, R) N, N'-Dihexanoyl-3,3'-dithiobis (2-aminopropanoic acid):

eq. A suspension of 3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester dihydrochloride in tetrahydrofuran (white liquid) was stirred and cooled to 0 ° C. 4 eq. Α-4-ethyldiisopropylamine and 2.2 eq. Hexanoyl chloride. The mixture was stirred in an ice bath for 4 hours and a white precipitate of N-ethyldiisopropylammonium chloride was filtered off. The solvent was removed by evaporation under reduced pressure and the crude material was redissolved in dichloromethane. After washing with water, the organic phase was dried over sodium sulfate. Filtration and evaporation of the solvent afforded the crude (R, R) - N, N'-dihexanoyl-3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester, which was recrystallized from methanol / water and triturated with diethyl ether.

(0.1 M) of the white dimethyl ester formed above and 0.5 M sodium hydroxide in 10% methanol was stirred vigorously at room temperature. After about 48 hours. the clear solution was raised to pH 2 and the resulting white crude precipitate was filtered and recrystallized from acetone / hexane to give the desired product as white crystals. Final yield: 28%.

Physical data:

Melting point = 132-135 ° C. [α] ² β ⁵ = -164 ° (C = 0.501, MeOH). 1 H-NMR (300 MHz, DMSO-d ₆ ): d 8.22 (2H, d, NH), 4.49 (2H, m, CHN), 3.14 (2H, dd, CH ₂ S), 2.91 (2H, dd, CH). ₂ S), 2.12 (4H, t, CH ₂ CO), 1.50 (4H, p, CH ₂ CH ₂ CO), 1.26 (8H, m (CH ₂ ) ₂ ), 0.87 (6H, t, CH ₂ ). H, 7.4; N, 6.4; S, 14.7. Found:% C, 49.4; H, 7.2; N, 6.2; S, 14.3.

An example. (R, R) - N, N'-Di (1-Oxo-octyl) -3,3 '-dithiobis (2-aminopropionic acid):

The compound was prepared according to the method described in Example 23 using octanoic acid chloride as the acylating agent.

The (R, R) -N, N'-di (1-oxo-octyl) -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester formed initially was recrystallized from ethyl acetate and the desired compound recrystallized from acetone / heptane and triturated with diethyl ether to give the product as white crystals. Final yield: 20%.

Physical data:

Melting point = 105-107 ° C. [<x] D ^{2 5} = -147 ° (C-0541, MeOH). 1 H-NMR (300 MHz, DMSO-d ₆ ): δ 8.18 (2H, d, NH), 4.48 (2H, m, CHN), 3.17 (2H, dd, CH ₂ S), 2.89 (2H, dd, CH). ₂ S), 2.12 (4H, t, CH ₂ CO), 1.49 (4H, m, CH ₂ CH ₂ CO), 1.26 (16H, m (CH ₂ ) ₂ ), 0.87 (6H, t, CH 2). Analysis. Calculated,%: C ^ H ^ N ^^ S ^ C, 53.6; H, 8.2; N, 5.7; S, 13.0. Found,%: C, 53.4; H, 8.5; N, 5.7; S, 12.6.

An example. (R, R) - N, N'-Di (1-Oxo-dodecanyl) -3,3'-dithiobis (2-amino-propanoic acid):

The compound was prepared according to the method described in Example 23 using dodecanoic acid chloride as the acylating agent. Hydrolysis was performed in 20% methanol.

The (R, R) -N, N'-di (1-oxo-dodecanyl) -3,3'-dithiobis (2-aminopropanoic acid) dimethyl ester formed initially was recrystallized from ethyl acetate. The title compound was recrystallized from a toluene / dichloromethane mixture to give the product as white crystals.

Final yield: 23%.

Physical data:

Melting point = 110-112 ° C. [α] D = -113 ° (C = 0.507, MeOH). 1 H-NMR (300 MHz, DMSO-d ₆ ): δ 8.19 (2H, d, NH), 4.48 (2H, m, CHN), 3.15 (2H, dd, CH ₂ S), 2.89 (2H, dd, CH). ₂ S), 2.11 (4H, t, CH ₂ CO), 1.49 (4H, m, CH ₂ CH ₂ CO), 1.25 (32H, m, (CH ₂ ) ₈ ), 0.87 (6H, t, CH ₃ ). . Analysis. Calculated,%: ^c 30 ^H 56 ^N 2 ° 6 ^S 2 ^: C, 59.6; H, 9.3; N, 4.6; S, 10.6. Found,%: C, 59.2; H, 9.4; N, 4.6; S, 10.3.

An example. (S, S) -N, N'-di (2-methylpropionyl) -3,3 '-dithiobis (2-aminopropanoic acid):

(17.6 g, 127 mmol) of potassium carbonate was dissolved in 40 mL of water and 40 mL of methylene chloride under nitrogen. The solution was cooled (-10 ° C) and added quickly

2-methylpropionyl chloride (5.6 mL, 40 mmol) and D-cysteine hydrochloride monohydrate (8.7 g, 49.5 mmol). The reaction mixture was stirred at room temperature for 4 hours, then hydrochloric acid was added until the pH of the mixture was less than 1. According to litmus paper, the aqueous layer was removed and light petroleum (40-60 °) was added to the organic layer, - D-cysteine precipitate.

N-Isobutyryl-D-cysteine (1 g, 5.2 mmol) was dissolved in 10 mL of methanol. Hydrogen peroxide (30%, 0.3 mL, 2.6 mmol) was added and the reaction mixture was stirred at room temperature for 6 h. Evaporation of the solvent under reduced pressure gave a white, crystallizing oil. Recrystallization from ethyl acetate gave the title compound as a white solid, which was dried in vacuo.

Yield: 0.55 g (55%)

Physical data: Melting point. = 135-137 ° C. 1 H-NMR (300 MHz, DMSO-d ₆ ): δ 8.16 (2H, d, NH), 4.47 (2H, m, CHN), 3.15 (2H, dd, CH ₂ S, J = 14 Hz, 5 Hz). , 2.93 (2H, dd, CH ₂ S, J = 14 Hz, 9 Hz), 2:43 (2H, h, CH (CH _{3) 2,} J = 7Hz), 1:01 (12H, d, CH _3, J = 7 Hz).

[α] ⁵ = θ + 167.6 (c = 0.516, MeOH).

An example.

The composition of A

Tablet containing 10 mg of active substance:

Active substance 10 mg

Lactose 100 mg

Potato starch 50mg

Polyvinylpyrrolidone 5 mg

Microcrystalline cellulose 15 mg

Magnesium stearate 1 mg

The composition of B

Direct compression tablet containing 5 mg of active ingredient:

Active substance 5 mg

Lactose, anhydrous 150 mg

Microcrystalline cellulose 50 mg

Colloidal silica 1 mg

Magnesium Stearate 2 mg

If necessary, the resulting tablets may be coated with a film, for example, copolymer of hydroxypropylmethylcellulose, hydroxypropylcellulose or dimethylaminoethyl methacrylate methacrylic acid ester.

Composition of C

Solution for injection, containing 1 mg / ml of active substance Active substance 1.0 mg

Sodium chloride 8.8 mg

Water for injections up to 1 ml

The composition of D

Oral solution containing 1 mg / ml of active substance

Active substance 1.0 mg

Sorbitol 150 mg

Glycerol 100 mg

Disodium edetate 0.5 mg

Condom as needed

Flavoring material as needed

Purified water to 1 ml

The composition of E

1 mg powder aerosol

The micronized active substance may be added to a powder inhaler such as R

Turbuhaler at a dose of 1 mg / dose.

Investigation of the effects of the compounds of the invention on a slow type hypersensitivity reaction in mice

The ability of the compounds of the invention to stimulate the immune response is illustrated by the slow type hypersensitivity (LTHR) response of mice.

Male Balb / C mice and females from Borihotsgaard (Denmark) and Charlie Rivers (England) weighing 18-20 g were used. The antigen used in these studies

4-Ethoxymethylene-2-phenyloxazolone (EFO), obtained from BSH (England).

Day. Mice were sensitized by subcutaneous injection of 150 ml of a 3% EFO solution in absolute alcohol / acetone (3: 1) in a mixture of shaved chest and abdomen. Oral treatment with this compound was initiated immediately after sensitization and continued for 6 days. Solvent (phosphate buffer, pH 7.0) was used as a positive control. On the seventh day after sensitization (day 6), both ears of both mice were treated by topical application of 20 ml of 1% EFO in olive oil. The thickness of the ear was measured before and 24 hours or 48 hours after application, using an Oditest spring instrument that measures the thickness of the skin fold. Affected and measured under pentabarbital anesthesia. The reaction intensity of LTHR was determined by the formula:

Tf 24/28 ^ t ° ' ^r ^ is expressed in mm, where and _t 24/48 ^ ^{is the} i i i i _t i o ^stor i o values in the individual test before exposure and after 24 and 48 hours, respectively. Data obtained are presented as mean values +/- scanning electron microscopy (SEM) The size of the scattering between the values of the groups was determined by the Student binary distribution criterion. Table 1 shows the results obtained after 24 and 48 hours, expressed as% increase in ear thickness compared to unaffected ear.

Table

I m u n o s t i m u 1 i a c i n i s act y v u m a s % increase, 24 or 0.03 3.0 mmol / kg % increase, 48 or 0.03 3.0 mmol / kg R R ² R ³ C (CH ₃ ) ₂ C (CH ₃ ) ₂ ch ₃ 110 ^XXX 92 ^XX 17 ^X 27 ^{XXX h c} 5 ^H 1 1 HC ₅ Hh ch ₃ 73 ^XX 86 ^XX 51 ^XXX , -XXX 45 hc ₇ h ₁₅ hc ₇ h ₁₅ ch ₃ 58 ^XXX 52 ^XXX 28 ^XX 20 ^X hc ₇ h ₁₅ hc ₇ h ₁₅ c ₂ h _{5 49} ^xxx 7th 0 0 COC (CH ₃ ) ₃ C (CH ₃ ) ₃ H 14 ^X 26 ^X 27 ^X COCH (CH ₃ ) ₂ CH (CH ₃ ) ₂ H 17th 18th 27 ^XX , .ΧΧΧ 02 coch ₃ C (CH ₃ ) ₃ H 24 ^X 26 ^X 14th 17 ^X H CH (CH ₃ ) ₂ H 37 ^XX 21XXX

x, xx, xxx: R <0.05, 0.01, 0.001

Claims (14)

DEFINITION OF INVENTION Compounds of the general formula COOR ³ I S - CH ₂ - CH - NH - R t I S - CH ₂ - CH - NH - CO - R ² C00R ³ wherein R is hydrogen or a group -CO-R ^ wherein R ^ is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, iso-propyl, 1-methylpropyl, tert. butyl, 3-methylbutyl or 2-methylbutyl, R is methyl, ethyl, n-propyl, n-butyl, n-pentyl, nhexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, isopropyl, o 1-methylpropyl, tert. butyl, 3-methylbutyl or 2-methylbutyl, and R is hydrogen, methyl, ethyl, propyl, isopropyl, butyl or isobutyl, provided 12 * that R and R are not simultaneously methyl, and further provided that when R ^J is present Is hydrogen, R and R are not simultaneously n-propyl or n-heptyl, or a physiologically acceptable salt thereof and / or a stereochemically isomeric form thereof. A compound according to claim 1 wherein R is hydrogen or -CO-R 1 wherein R 4 is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n -octyl, n-nonyl, n-decyl, n-undecyl, iso-propyl, 2 1-methylpropyl, tert. butyl, 3-methylbutyl or 2-methylbutyl, R is methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n- undecyl, iso-propyl, 1-methylpropyl, tert. butyl, 3 3-methylbutyl or 2-methylbutyl, and R is hydrogen, methyl, ethyl, with 12 -> provided that R and R are not simultaneously methyl and further provided that when R ³ 1 2 is hydrogen, R and R are not simultaneously n-propyl or n-heptyl. 1? 3. A compound according to claim 1 wherein R and R are isopropyl and R is methyl. 1 2 4. A compound according to claim 1 wherein R and R are o n-pentyl and R ³ is methyl. 1 2 5. A compound according to claim 1 wherein R and R are n-heptyl and R is methyl or ethyl. 1 2 6. A compound according to claim 1 wherein R and R are simultaneously isopropyl and R is hydrogen. 1 2 7. The compound of claim 1 wherein R and R are simultaneously tertiary. butyl, and R is hydrogen. 8. A process for the preparation of a compound of general formula I as claimed in claim 1, characterized in that a) oxidizes the N-acylcysteine derivative of the formula COOR ³ HS CH ₂ - CH - NH - CO - R \ 1 3 3 wherein R and R are as defined above or when R is hydrogen, an alkali salt thereof is chosen and the compound of formula COOR ³ I ch ₂ - 1 -CH-NH - - CO ch ₂ - - CH - NH - I - CO 1 COOR ³ or b) carrying out the reaction in the presence of an appropriate base of a compound of the formula COOR ³ CH ₂ - 1 CH - NH ₂ ch ₂ - CH - NH ₂ 1 COOR ³ or a salt thereof, wherein R is as described above, with a compound of the formula R ¹ is COX, wherein R is as described above, and COX is a reactive group capable of reacting with an amino group to form an amide group to form a compound of formula COOR ³ I ch ₂ - 1 CH - NH - CO ch ₂ - CH - NH - CO 1 COOR ³ or c) carrying out the reaction of 2 - (N-acylamino) - 3 - halopropanoic acid derivative of the formula COOR ³¹ I Y - CH ₂ - CH - NH - CO - R ¹ , 13I 3 wherein R is as described above, R is R as described above, or an acidic or basic labile organic group and Y is a halogen atom with a sulfuric or disulfide dianion in the presence of a base to form a compound of formula COOR ³ | ch ₂ - 1 -CH-NH - - CO ch ₂ - - CH - NH | - CO 1 COOR ³ or d) oxidizing a mixture of N-acylcysteine ester derivatives of the formulas COOR ³ I HS - CH ₂ - CH - NH - CO - R ¹ and COOR ³ I HS - CH ₂ - CH - NH - CO - R ² 12 3 3 wherein R, R and R are as described above or when R is hydrogen, an alkali salt thereof is chosen to form a compound of formula COOR ³ I S - CH ₂ - CH - NH - CO - R ¹ I S - CH ₂ - CH - NH - CO - R ² I COOR ³ or e) oxidizing a mixture of cysteine or a cysteine ester and an N-acylcysteine derivative of the formulas COOR ³ I HS - CH ₂ - CH - NH ₂ or a salt thereof and COOR ³ I HS - CH ₂ - CH - NH - CO - R ¹ 13 or its alkaline or hydrochloride salts, wherein R and R are as described above, to form a compound of formula S I S COOR ³ | ch ₂ ch ₂ - CH - NH - CH - NH I COOR ³ co R ¹ or a salt thereof, or (f) carrying out the reaction in the presence of a suitable base and in the presence of an excess of the cysteine or cysteine diester formula COOR ³ I S - CH ₂ - CH - NH ₂ I S - CH ₂ - CH - NH ₂ I COOR ³ or a salt thereof, wherein R is as described above, with a compound of the formula R ² is COX, wherein R and COX are as defined above, to form a compound of formula or COOR ³ I S - CH ₂ - CH - NH ₂ S - CH ₂ - CH - NH - CO - R ² I COOR ³ g) carrying out the reaction in the presence of a suitable base of a compound of the formula COOR ³ I S - CH ₂ - CH - NH - CO - R ¹ I S - CH ₂ - CH - NH ₂ COOR ³ or a salt thereof, wherein R and R are as described above, with a compound of the formula R ² is COX wherein R and COX are as described above for the compound of formula COOR ³ I S - CH ₂ - CH - NH - CO - R ¹ I S - CH ₂ - CH - NH - CO - R ² I COOR ³ h) reacting the N-acylcysteine derivative of the formula COOR ³ I HS - CH ₂ - CH - NH - CO - R ¹ or its salts, Wherein R and R are as described above, with the activating reagent to form the adduct COOC ₂ H ₅ COOR ³ I I NS - CH ₂ - CH - NH - CO - R ¹ I NH I Further reaction of COOC ₂ H ₅ with a second, different or same N-acylcysteine or cysteine ester derivative gives the compound of formula COOR ³ I S - CH ₂ - CH - NH - CO - R ¹ I S - CH ₂ - CH - NH - CO - R ² I COOR ³ i) carries out the reaction of the compound formula COOR ³¹ I R ⁴ - S - CH ₂ - CH - NH - CO - R ¹ o coor ³¹ -s-ch ₂ -ch-nh-cor ¹ II and R ¹ is as described above, with a compound of formula COOR ³¹ H - S - CH ₂ - CH - NH - CO - R ² wherein R is as described above for the compound of formula S I S COOR ³¹ I ch ₂ - I -CH-NH - -CO-R ¹ ch ₂ - - CH - NH - i -CO-R ^{2 1} , 1 COOR ³ 3 then removes the protecting group R, if R = H, or j) esterifying the compound of the formula COX ¹ that is ch ₂ I - CH - NH - CO ch ₂ - CH - I NH - CO I cox ¹ Wherein R and R are as described above and X is OH or halogen, by a compound of formula R ^3E - OH, 3E wherein R is methyl, ethyl, propyl, isopropyl, butyl or isobutyl, to form a compound of formula coor ^3E 1 S - ch ₂ I 1 —CH — NH - R 1 s - ch ₂ - CH - NH 1 CO - R ² 1 COOR ^3E or k) alkylating the compound of formula COO ^(_) | s - ch ₂ 1 1 - CH - NH - R 1 s - ch ₂ - CH - NH 1 COO ^(_) CO - R ² is as described above, with a compound of the formula wherein R and R R ^3E - Z, 3E wherein R is as described above and Z is halogen, alkylsulfate, tosylate or other nucleophile, to form a compound of formula coor ^3E I S - I ch ₂ l - CH - NH - R 1 s - ch ₂ - CH - NH 1 coor ^3E - CO - R ² 1) Reacts in the presence of a suitable base with the formula of a cystine derivative with a protected carboxyl group CH ₂ - COOR ^3p I CH - NH CH ₂ - CH - I NH; 1 COOR 3P 3P or its hydrochloride salt, wherein R is an acidic or basic labile organic group, with a compound of the formula R ¹ is COX, wherein R ¹ is as described above, and COX is a reactive group capable of reacting 3P with an amino group to form an amide group, then removing the protecting group R to form a compound of formula COOH ch ₂ - CH - NH - CO ch ₂ - CH 1 COOH NH - CO m) Reacts under alkaline conditions, formulas of a cystine derivative with a protected carboxyl group coor ^3P t CH ₂ - 1 CH - NH ₂ CH ₂ - CH - NH ₂ 1 coor ^3P 3P or its hydrochloride salt, wherein R is as described above, with a compound of the formula R - COX, 2. . . . 3P wherein R and COX are as described above and then removes the protecting group R to form a compound of formula COOH I S - CH ₂ - CH - NH ₂ I ₂ S - CH ₂ - CH - NH - CO - R ² I COOH n) reacting a cysteine derivative with a protected carboxyl group of the formula coor ^3P I S - CH ₂ - CH - NH ₂ I S - CH ₂ - CH - NH - CO - R ² I C00R ^3P 2 3P or its hydrochloride salt, wherein R and R are as described above, with a compound of the formula R ¹ - COX wherein R ¹ and COX are as described above and then removes the protecting group 3P R, to form a compound of formula COOH I ch ₂ I - CH - NH - CO ch ₂ - CH - I NH - CO I COOH (o) Balancing the formulas of a mixture of cysteine derivative and cysteine derivatives in an alkaline aqueous solution COOH I CH ₂ I - CH - NH - CO ch ₂ - CH - NH - CO 1 COOH R ¹ COOR ⁴ I HS - CH ₂ - CH - NH - CO - R ² 1 2 or their alkaline salts, wherein R and R are as described above to form a compound of the formula COOH I ch ₂ 1 - CH - NH - CO ch ₂ - CH | NH - CO COOH and finally, if desired, converting the compounds of any one of (a) to (o) into a physiologically acceptable salt. 9. A pharmaceutical preparation comprising as active ingredient a compound according to any one of claims 1 to 7. Pharmaceutical preparation according to claim 9, characterized in that it is in dosage form. Pharmaceutical preparation according to claims 9 and 10, characterized in that it contains the active substance together with a pharmaceutically acceptable carrier. A compound according to any one of claims 1 to 7 for use as a therapeutically active substance. 13. A process for the preparation of an immunomodulatory drug comprising the use of compounds according to any one of claims 1 to 7. A compound according to any one of claims 1 to 7 for use in the treatment of diseases associated with immune defects in a mammal, including a human.