FI66366C - OIL FRAMSTAELLNING AV SOM LAEKEMEDEL ANVAENDA 1,4-THIAZINE-3-CARBOXYL SYRADERIVAT - Google Patents

Description

1 66366

The present invention relates to a process for the preparation of 1,4-thiazine-3-carboxylic acid derivatives which can be used as medicaments for the preparation of 1,4-thiazine-3-carboxylic acid derivatives.

For the preparation of 1,4-thiazine-3-carboxylic acid derivatives of formula I

<?> n R1 (I) R S_R3 R5 R4 °

where J

R 1 and R 2, which may be the same or different, represent hydrogen, a straight or branched C 1-4, C 2, C 3 or C 4 alkyl radical or phenyl optionally substituted by a halogen atom, such as fluorine, chlorine or bromine, straight or with a branched C 1-4, C 2 -C 3 or C 4 alkyl radical, or a straight or branched C 1-4, C 2, C 3 or C 4 alkoxy radical, R 1 and R 2 may form a cycloalkyl with an adjacent carbon atom having 3, 4, 5, 6, 7 or 8 carbon atoms, wherein one or more carbon atoms may be optionally replaced by a sulfur or oxygen atom or a group SO, SO 2 or NR 9, wherein

Ro represents a hydrogen, straight or branched C 1-4, C 0 -C 3 or C 4 alkyl radical, phenyl or a benzyl group, R 3 represents: - a hydroxyl group, 2 66366 <* t - an ORg group in which Rg represents a straight or branched C 2-, C 1-4 C 8, C 8, C 8, C 8, C 8 -C 4, H 2 or C 12 alkyl, O-CH-COOR or OCH -OCOR 4, R 10 p 10 wherein R 1 q represents hydrogen or a C 1 -C 4 or C 2 alkyl radical and R 8 - represents a straight or branched C 1 -C 4, C 2, C 8, C 4, C 8, C 8, C 7, C 7 -, C 8, C 8, C 10, C 1-4 or C 12 alkyl radicals, R 12 - groups N, in which R 2 and R 13 »which may be the same or different, denote hydrogen or straight or branched A C 1-4, C 2-, C 3- or C 1-4 alkyl radical, or a -3-phthalidyloxy group or a 1- (2,5-dioxopyrrolidinyl) -1-ethoxy group, R 4 represents a hydrogen atom, a straight or branched C 1-4, C 2-, A C3, C4, C8, C8, C7 or C8 alkyl radical, a straight or branched C1-4, C2, C8, C4, C8, C8, C7 or C8 acyl group or a group CONH2 , R 3 and R 4 may form a hydantoin ring with adjacent carbon and nitrogen atoms, R 5 and R 6, which may be the same or different, represent hydrogen, straight or branched C 1, C 2, C 8, C 4, C 8, C 8, A C7 or C8 alkyl radical, phenyl optionally substituted by a halogen tower such as fluorine, chlorine or bromine, a straight or branched C1-, C2-, C8- or C1-4 alkyl radical or a straight or branched C1-8, C2-, C8- or a C 1-4 alkoxy radical, or a group C 1-6 COOR 4 in which hydrogen represents or a straight or branched C 1-4, C 1-4, C 1-4 or C 1-4 alkyl radical, Ry denotes hydrogen , a straight or branched C 1-4, C 1 -C 4 or C 1-4 alkyl radical, a geminal dimethyl radical or phenyl optionally substituted by a halogen atom, such as fluorine, chlorine or bromine, a straight or branched C 1-4, or C 1-4 alkyl radical, an alkyl radical or a straight or branched C 1-4, C 1-4 C 1-4 alkoxy radical, and

f U

n can have the value 0, 1 or 2.

If R 1, R 2, R 2 and R 2 represent hydrogen and if R 2 represents a hydroxyl group or a straight or branched C 1 -C 4 alkyl radical, R 1. or R 1 does not represent hydrogen, straight or branched C 1 -C 4 alkyl radical or phenyl.

According to a preferred aspect of the invention, the invention relates to compounds of formula I, wherein: R 1 and R 2, which may be the same or different, represent hydrogen or a straight or branched C 1 -C 4 alkyl radical, R 1 represents a hydroxyl group, a group R 2 represents a straight or branched C 1 -C 4 alkyl group, a group O-CH-COOR ^ R 10 or a group O-CH-OCO 4, in which R 1 represents a hydrogen or a methyl-R 10 radical and R 4 represents a straight or branched A C 1 -C 6 alkyl radical, R 4 represents hydrogen or a methyl group, R 5 and R 6 * may be the same or different, denote hydrogen, a straight or branched C 1 -C 4 alkyl group or a group C 1 -C 4 OR 4 in which R 1 represents hydrogen or a C 1 -C 4 alkyl group, R 1 represents hydrogen and n may have a value of 0 or 1.

4 66366

A preferred group of compounds of formula I is that wherein R 1 and R 6 represent hydrogen or a methyl group, R 6 represents a hydroxyl group or a group OR 8, wherein R 8 represents a straight or branched C 1 -C 4 alkyl group, R 4 represents hydrogen or a methyl group, R 4 represents a hydrogen or methyl group, hydrogen and R 1 represents a straight or branched C 1 -C 4 alkyl group, R 2 represents hydrogen and n may have a value of 0 or 1,

Examples of compounds according to the invention are: 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylic acid, ethyl-1-oxo-2,2,5-trimethyl-1,4-thiazine -3-carboxylate, 2,2,4,5-tetramethyl-1,4, -thiazine-3-carboxylic acid, 1- (pivaloyloxy) ethyl-1-oxo-2,2,5-trimethyl-1,4-thiazine -3-carboxylate, (octyloxycarbonyl) methyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 1- (ethoxycarbonyl) ethyl 1-oxo-2,2,5-trimethyl -1,4-thiazine-3-carboxylate, 5-butyl-2,2-dimethyl-1,4-thiazine-3-carboxylic acid, 2,2-dimethyl-5-isopropyl-1,4-thiazine -3-carboxylic acid, 5-methyl-2-phenyl-1,4-thiazine-3-carboxylic acid and ethyl 3-carboxy-2,2-dimethyl-1,4-thiazine-5-acetate.

If the derivatives of the formula I are in the form of acid addition salts, they can be converted into their free bases or salts of other acids by conventional methods.

The most commonly used salts are pharmaceutically acceptable, non-toxic acid addition salts formed with suitable inorganic acids, for example hydrochloric acid, sulfuric acid or phosphoric acid, or suitable organic acids, such as aliphatic, cycloaliphatic, aromatic, araliphatic or carbocyclic or heterocyclic or heterocyclic. , vinegar, propion, amber, glycol, glucone, milk, apple, wine, lemon, ascorbine, glucuron, maleic, fumaric, fire grape, aeparagine, glutamine, benzoic , anthranilil, hydroxybenzoic, salicylic, phenylacetic, almond, embonic, methanesulfonic, ethanesulfonic, pantothenic, toluenesulfonic, sulphanilic, cyclohexylaminosulfonic, stearic, kt 6 6 3 6 6 algin -, (3-hydroxypropionic, β-hydroxybutyric, oxalic, malonic, galactaric and galacturonic acids. These salts can also be derived from natural or synthetic amino acids such as lysine, g lysine, arginine, ornithine, asparagine, glutamine, alanine, valine, threonine, serine, leucine, cysteine and the like.

When OH represents, the present invention also encompasses salts formed between a carboxylic acid group thus present in Formula I and bases such as sodium hydroxide, potassium hydroxide, lithium hydroxide, ammonium hydroxide, magnesium hydroxide and calcium hydroxide, and also internal salts. (zwitterions).

The compounds of formula I may have one or more centers of asymmetry and may exist in the form of optical isomers, racemates or diastereoisomers; all of these forms are part of the present invention. Optical isomers can be obtained by stereospecific or stereoselective syntheses or by resolution of racemates by conventional methods, for example by reaction with optically active acids such as tartaric, diacetyltartaric, tartranilic, tartranilic , and then releasing the optically active bases from these salts. Optically active compounds of formula I can also be obtained using optically active starting materials. Mixtures of diastereoisomers can be separated in the same manner as the diastereoisomeric salts mentioned above.

Arterial and venous thrombotic diseases are generally just as important a cause of mortality and especially a cause of morbidity in Western countries. The products of the invention allow a new therapeutic approach, which differs from the approach with compounds that affect the functions of blood particles on the one hand and with the approach with fibrinolytic enzymes on the other hand. The increase in blood fibrinolytic activity resulting from the increase in the synthesis and / or release of natural activators of fibrinolysis as a result of oral or parenteral administration of the compounds of the invention makes it possible to envision important therapeutic uses under better safety conditions. Thus, severe venous thrombosis and its often fatal consequences, namely pulmonary embolism, could be prevented by oral treatment before risky surgical interventions and in the weeks thereafter.

It is also possible to imagine the prevention of vascular thrombosis when it occurs as a complication of inflammation of primary or secondary superficial blood vessels during a period of time in which patients are immobile for a long time. The products of the invention may also be used in conditions where a decrease in blood fibrinolytic activity occurs, such as after surgery, during general anesthesia or in the case of insulin-treated diabetes. Finally, products of the invention - either alone or in combination with compounds that inhibit platelet adhesion and aggregation - are presented for the prevention of recurrent arterial or venous thrombotic seizures. In fact, it is known that in patients who have had a cerebrovascular or coronary artery occlusion (myocardial infarction), the risk of recurrence of the blockage justifies the use of prophylactic treatment.

The products of the invention work by oral administration, which makes them easy to use, and can be administered over a long period of time, in contrast to current fibrinolytic compounds, because they have no enzymatic activity and are not immunogenic and act through activators normally present in the body. Their use alone or in combination with antiplatelet compounds provides a way to treat and prevent arterial and venous thrombosis.

Also included in this invention are pharmaceutical compositions comprising as active ingredient at least one compound of general formula I and / or one of its salts with a pharmaceutical carrier. These compositions are presented so that they can be administered orally, rectally or parenterally.

Thus, for example, compositions for oral administration may be liquid or solid and may take the form of tablets, capsules, granules, powders, syrups or suspensions; such compositions include additives and carriers commonly used in galenic pharmacy, inert diluents, disintegrating agents, binders and lubricants such as lactose, starch, talc, gelatin, stearic acid, silicic acid, magnesium stearate, magnesium stearate, calyvinylpyrrolidone, calyvinylpyrrolidone,

Such formulations may be prepared by prolonging the degradation and thus the duration of action of the active ingredient.

Aqueous suspensions, emulsions, and oily solutions are prepared using sweetening agents, for example, dextrose or glycerol, and flavoring agents, for example, vanillin, and may also contain thickening agents, wetting agents and preservatives.

oil emulsions and solutions · are formulated in vegetable or animal oils and may contain emulsifiers, flavor enhancers, dispersants, sweeteners and antioxidants.

Sterile water, aqueous polyvinylpyrrolidone, peanut oil, ethyl oleate or the like are used as a medium for parenteral administration. These aqueous or oily spray solutions may contain thickeners, wetting agents, dispersing agents and gelling agents.

According to the first procedure, the vinyl thioether of formula II obtained from A.I. Virtanen et al. in Acta Chem. Scan., 1966, 20, 1163-1185, is cyclized to give derivative I according to the following scheme: R, COR ,.

? 1 / C0-R3 r / / R2-C CH 2 —- + - (λ "· ->» v ·

y ~ / H

76 R7 R6

II I

R 1, R 2, R 3, R 4, R 5, R p, R y and n are as defined above.

The cyclization is preferably carried out in a basic medium. The base may be organic, such as triethylamine, pyridine or Ν, Ν-dimethylaniline, or inorganic, such as, for example, alkali metal hydroxides, such as sodium hydroxide or potassium hydroxide, or also ammonium hydroxide.

If the base used is an organic base, the solvent is preferably an inert organic solvent such as chlorinated solvents such as chloroform or methylene chloride, aromatic or aliphatic hydrocarbons such as benzene, toluene or various petroleum ether fractions, or also acetonitrile, N, N, 66-dimethyl or solvents such as dimethyl sulfoxide.

If the base is inorganic, the solvent used is preferably water, and the concentration of the basic solution is 0.01 N to 10 N, and preferably 0.2 N to 2 N. The reaction takes place at a temperature between 0 ° C and the reflux temperature of the solvent.

However, it may be very advantageous to carry out the reaction at a temperature close to ambient temperature, especially if the reacting compounds or products predominantly contain a stereoisomer which is likely to be isomerized if the conditions used are too strong.

Another procedure involves cyclization with cysteine derivative III obtained from J.F. Carsonin et al. in J. Org. Chem. 29, 2203 (1964), according to the following scheme:

R? ✓ / COR * «I C0R

(H) n (s) X NHR - (-) (0) n -5 N-R4 CH-C-R. In this scheme, R 1, R 2, R 2, R 2, R 2, R 2, R 2 and n are as defined above and X represents a halogen atom such as chlorine, bromine or iodine, a hydroxyl group or a group which can be easily removed, such as a tosyl or mesyl group.

If the cyclization is carried out in a basic medium, the conditions described for the first method can be applied here. If the cyclization is carried out in an acidic medium, then an inorganic or organic acid such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, formic acid or acetic acid is used.

10 6 6 3 6 6

It is also possible to use dehydrating agents such as phosphorus pentoxide, concentrated sulfuric acid or a carbodiimide such as dicyclohexylcarbodiimide.

These reactions are usually carried out in inert organic solvents, such as chlorinated solvents such as chloroform or methylene chloride, benzene, toluene, xylene, chlorobenzene, dichlorobenzene or petroleum ether, dimethylform, dimethylform, or dimethyl sulfoxides, or aromatic hydrocarbons. The reaction takes place at a temperature between ambient temperature and the reflux temperature of the selected solvent.

According to another procedure, the cysteine derivative IV is cyclized by reaction with α-haloketone V; this gives the unsaturated cyclic compound VI, which is then reduced to give the derivative VII according to the following scheme:? i '

HS-C - CH - C0R3 ♦ Y-CH-C0-R5 f K

R2 NH2 R7

IV V VI

«

Reduction VSi VII I ^

B

11 66366 t

R 1, R 2, R 2, R 2 and R 7 are as defined above and Y represents a halogen atom such as chlorine, bromine or iodine.

The first step of the process takes place in an inert organic solvent such as alcohols such as methanol, ethanol or propanol, ethers such as dioxane or tetrahydrofuran, solvents such as dimethyl sulfoxide or Ν, Ν-dimethylformamide or aromatic hydrocarbons such as benzene or toluene.

# •

Bases which can be used are alkali metal hydroxides, such as sodium hydroxide or potassium hydroxide, or alkali metal alcoholates, such as, for example, sodium methanolate or potassium methanolate, sodium ethanol or potassium methanolate or sodium t-butanolate or potassium t-butanolate.

The reaction takes place at a temperature between -20 ° C and the reflux temperature of the solvent; it is preferably carried out at a temperature between 0 ° C and ambient temperature.

of*.

A dehydrating agent, such as a molecular sieve, may preferably be added to the reaction mixture.

N 1 r

The derivative VI can be isolated and purified, or alternatively the crude mixture can be used directly for the reduction step.

The imine VI is reduced in the presence of hydrogen and a reduction catalyst such as platinum, platinum oxide or palladium on carbon in a solvent such as methanol, ethanol, ethyl acetate or glacial acetic acid under general pressure and more preferably at higher pressure, or with an alkali metal hydride such as sodium hydride such as sodium or lithium aluminum hydride in a solvent such as ether or tetrahydrofuran.

*

As a general rule, this reduction takes place at ambient temperature, but depending on the reactivity of the system, it may sometimes be advantageous to heat or cool the reaction mixture.

An acid such as hydrochloric acid, sulfuric acid or acetic acid can sometimes catalyze the reaction.

To obtain sulfoxides, it is possible to use substances such as iodine, bromine in water or in the presence of acetate ions or also complexed with pyridine, peracids such as peracetic, monoperphthalic or m-chloroperbenzoic acids, N-halosuccinimides such as N-brornsuccinimide, sodium chloride, hypochlorite or t-butyl or i-propyl hypochlorite, periodates such as sodium periodate, hydrogen peroxide in the presence of acetic anhydride or vanadium pentoxide in t-butanol, nitrates such as acetyl nitrate or cerium ammonium nitrate, oxides, nitric pentoxide, peroxides, ozone, oxygen in mono- or tri-state, acids such as nitric acid, chromic acid or Caron's acid, or also other substances such as sulfuryl chloride, 1-chlorobenzotriazole, chloramine, N-chloronylenodibenzodetabenzodiazide dichloride, iodobenzene dichloride, , N-chlorotriazole, 2,4,4,6-tetrabromocyclohexadienone or chloroacetic acid (HAuCl 2).

These oxidation reactions are performed in solvents such as water, acetic acid, chloroform, methyl bichloride, carbon tetrachloride, methanol, t-butanal or acetone.

In order to obtain sulfones, it is preferable to use substances such as hydrogen peroxide, preferably in the presence of zirconium salts, peracids such as peracetic, monoperphthalic and m-chlorobenzoic acids ( , selenium oxide, t-butyltypochlorite, nitric acid, ozone, oxygen, preferably in the presence of iridium and rhodium salts, iodobenzene-66366 dichloride or periodic acid, or use electrochemical oxidation. These reactions are performed in solvents such as water, acetic acid, chloroform, methylene chloride, methanol, ethanol, isopropanol, t-butanol, dioxane or acetone.

In certain cases, it may be advantageous to carry out the oxidation reactions described above with compounds in which the groups sensitive to the oxidizing agent used are protected.

The cyclization, reduction and oxidation steps described below may also form part of a continuous process that does not require the isolation of any intermediate.

It is noteworthy that the conversion of a derivative I with n = 0 to a derivative I with n = 1 or 2 can be performed at any step in the production of the 1,4-thiazine ring.

When a substituent denotes a straight or branched C 1 -C 6 alkyl group or a straight or branched C 1 -C 4 acyl group, it is obtained by alkylation or acylation or acylation of an amine according to the following scheme:: "VS NH ♦ -" " '"V! V * 4

VIII IX

R 1, R 2, R 4, R 5, R 2, R 6 and n are as defined above.

Z represents a halogen atom such as chlorine, bromine or iodine or a group which can be easily removed, such as 14,6366 tosyl or mesyl groups.

The reaction is preferably carried out in an inert organic solvent such as chlorinated hydrocarbons such as chloroform or methylene chloride, aromatic or aliphatic hydrocarbons such as benzene, toluene or petroleum ether, alcohols such as methanol and ethanol or also acetonitrile or ether. The temperature is between ambient temperature and the reflux temperature of the reaction mixture. The reaction may preferably be carried out in the presence of an organic base such as pyridine, triethylamine or N, N-dimethylaniline, an inorganic base such as alkali metal or alkaline earth metal hydroxides, carbonates and bicarbonates or finely divided lime.

Alkylated derivatives can also be obtained by reduction of the corresponding acylated derivative according to conventional methods.

The substituent = CONH2 is conventionally obtained by reacting derivative VIII with an alkali metal isocyanate or ammonium isocyanate according to the following scheme:

i I

V / C0R3 R1 fOR3 h7 ~ \ r27 \ (°) n- ^ NH + MNCO-- (°) n'S N-CONH2) ^ Rs yv ~ R5 K7 R6 R7 r6 5

VIII X

In this scheme, R 1, R 2, R 1, R 2, R 2, R 1 and n are as defined above, and M represents a monovalent cation such as sodium, potassium, lithium or ammonium ion.

The reaction is carried out in a general manner in water at a temperature between normal temperature and reflux temperature.

ΐ · 15 66366

Hydantoin XI is obtained by heating a derivative X in which R 1 represents a hydroxyl group in an aqueous acid solution.

0/1 / C0R3 / 1/27 \ h * (0) n-S N-CONH2- / 1 ^ «5 V-7 <° * 7« 5 * 7 »s 5

X XI

In this scheme, R 1, R 2, R 1, R 5, R 8, R 7 and n are as defined above. The acid used is a mineral acid, such as halogen-containing hydro acids such as hydrochloric acid, hydrobromic acid or hydroiodic acid, or nitric acid, sulfuric acid or also phosphoric acid.

The reaction mixture is usually heated to a temperature close to the reflux temperature of the solution.

Conversion of derivatives in which R 1 represents a hydroxyl group to derivatives in which R 2 represents an OCHCOOR ^ • * 10 or OCH-OCOR 4 is an esterification reaction. Acid esterification R10 is a very common reaction that can occur in a number of ways. Conventionally, the acid and alcohol are reacted in the presence of an acid catalyst such as hydrochloric acid, sulfuric acid or p-toluenesulfonic acid. This reaction is preferably carried out under anhydrous conditions, and one of the reactants is used in large excess. The solvent may be either one of the reactants or an inert organic solvent such as chlorinated hydrocarbons such as chloroform or carbon tetrachloride or aromatic or aliphatic hydrocarbons such as benzene, toluene or petroleum ether. The temperature is between the normal temperature and the reflux temperature of the reaction mixture.

Another procedure involves distilling the water as soon as it is formed, using suitable equipment. The reaction conditions are exactly the same as those described above, except for the fact that it is not necessary to use one of the reactants in large excess.

The hydrolysis of the ester is carried out under conditions similar to the esterification reaction, but in this case one of the reactants, namely water, is used in very large excess.

The catalysis and temperature conditions are the same as in the esterification.

Derivatives in which R 1 represents a hydroxyl group or groups such as ORq, OCHCOQR-. or OCH-OCO.R, converting to i 11, 11 R 10 R 10 "R 12 derivatives in which R 1 represents a group N. is r 15 is a conventional reaction which is very well documented in chemistry and which can be carried out according to several methods described below.

For example, a carboxylic acid can be reacted with an amine, and pyrolysis of the salt thus formed, as well as the action of a dehydrating agent such as P 2 O 2, results in the formation of an amide.

Another method involves the conversion of a carboxylic acid to an acid halide followed by reaction with an amine to an amide.

The conversion of an acid to an acid halide is often carried out without a solvent using thionyl chloride, phosphofripentachloride or phosphorus oxychloride. Corresponding bromides can also be used. To complete the reaction, it is often necessary to heat the reaction mixture to a temperature between 50 ° C and 150 ° C. If a solvent is necessary to carry out the reaction, this is an inert organic solvent such as hydrocarbons such as benzene, toluene or petroleum ether or ethers such as diethyl ether.

The reaction of the acid halide with the amine is performed by cooling the reaction mixture to a temperature between 0 ° C and -50 ° C using an excess of amine (at least 2 equivalents or at least 1 equivalent of amine and at least 1 equivalent of a tertiary organic base such as triethylamine). Conventionally, the acid chloride is added to a solution of the amine in an inert organic solvent, as defined above, or also in water.

Another procedure involves the reaction of a carboxylic acid with an amine in the presence of a coupling reagent such as used in peptide syntheses. Today, there are a huge number of coupling reagents, such as dicyclohexylcarbodiimide, N-ethyl-N'-3-dimethylaminopropylcarbodiimide, phosphines, phosphites, silicon tetrachloride or titanium tetrachloride or EEDQ.

The amination of the ester is usually carried out either in water or in an inert organic solvent. Examples of solvents which may be used are aromatic hydrocarbons, such as benzene or toluene, aliphatic hydrocarbons, such as hexane or petroleum ether, or halogenated hydrocarbons, such as methylene chloride or chloroform.

The presence of a strong base may be necessary in the reaction with amines of low basicity or sterically hindered amines. The above reaction can be carried out at a temperature between ambient temperature and the reflux temperature of the solvent.

Depending on the conditions used, hydrolysis of an amide, namely a derivative having a group N - R 2 *, may result in the formation of a carboxylic acid or esters R 13 18 66366.

By all the methods described above, the 1,4-thiazine derivatives according to the invention can be easily obtained. However, the composition of the resulting mixture of diastereoisomers varies depending on the method used. The fractionation of these diastereoisomeric mixtures is carried out in a manner commonly used in organic chemistry.

Detailed examples of the preparation of some of the derivatives of the invention are given below. The purpose of these examples is in particular to give a more detailed description of the detailed characteristics of the methods according to the invention.

Example 1. 2,2-Dimethyl-1,4-thiazine-3-carboxylic acid 1) 5- (2-hydroxyethyl) penicillamine

A solution of 17.5 g (0.14 mol) of 2-bromoethanol in 100 ml of ethanol is added dropwise, with good stirring, to a solution of 14.9 g (0.10 mol) of penicillamine in 75 ml of 2N sodium hydroxide. aqueous solution kept under an inert atmosphere (N 2) · When the addition is complete, add ethanol in an amount sufficient to obtain a homogeneous medium, followed by stirring at ambient temperature for 24 hours. After neutralization with concentrated hydrochloric acid, most of the alcohol is evaporated off under reduced pressure and the volume is made up to 300 ml with water. The resulting solution is applied to a Dowex 50 type resin column in acid form. The resin is washed with water and then eluted with 3 liters of 3 N ammonia solution. The eluate is evaporated to dryness. The residue is treated with 50 ml of water and 500 ml of ethanol. The solid is then filtered off and recrystallized from isopropanol. Is obtained. 9.0 g (47%) of a white solid. Melting point: 148-150 ° C.

19 66366 2) S- (2-chloroethyl) penicillamine (hydrochloride)

A mixture of 9.0 g (47 mmol) of the above product and 350 ml of 38% hydrochloric acid is heated for 7 hours in a bath maintained at 95 ° C.

The mixture is evaporated to dryness under reduced pressure and the residue is recrystallized from isopropanol. 8.9 g (78%) of product are obtained, which is homogeneous as determined by thin-layer chromatography on silica (eluent BuQH / AcOH / t = 0.13: 3: 5). Rf: 0.59.

3) 2,2-dimethyl-1,4-thiazine-3-carboxylic acid 6.7 g (27 mmol) of the above chloride are dissolved in 450 ml of N, N-dimethylformamide, then 50 ml of triethylamine are added and the mixture is heated 2, 5 hours in a bath maintained at 95 ° C. The mixture is then evaporated to dryness under reduced pressure. The residue is dissolved in 75% water and the solution is filtered into 250 ml of Dou / ex 50 resin. The resin is washed with water and then eluted with 1.5 l of ammonia solution. The eluate is concentrated under reduced pressure to a volume of 400 ml, the resulting solution is then treated overnight with 100 ml of Amberlite IRC 50 type resin in acid form. After the resin is filtered off, the solution is concentrated to a volume of 30 ml. The crystals that have appeared are filtered off and redissolved in 10 ml of water. Gradual addition of 40 ml of acetone causes reprecipitation of the final product. Weight: 3.1 g (66 S). Melting point: 320 ° C (decomposition).

* IR, NMR and mass spectra are consistent with the expected structure

Elemental analysis: C H N

% calculated 48.0 7.5 8.0% found 47.9 7.4 7.9 66366 20

Example 2. 2,2,5- (R, S) -Trimethyl-1,4-thiazine-3- (S) -carboxylic acid 1) 5-Allyl-penicillamine

A solution of 6.6 g (4.1 ml; 55 mmol) of allyl bromide in 50 ml of ethanol and 35 ml of 2N aqueous sodium hydroxide solution are slowly and simultaneously added to a suspension of 7.5 g (50 mmol). penicillamine in 50 ml of ethanol, keeping the temperature of the reaction mixture below 20 ° C.

When the addition is complete, the mixture is stirred for the next about 2 hours while testing for the disappearance of the starting ethiol (silica thin layer; eluent: BuOH / AcOH / H 2 O: 12: 3: 5; development with ninhydrin). The solution is neutralized with concentrated hydrochloric acid and then concentrated until a solid appears. The volume is made up to 300 ml with water and the solution obtained is filtered on a sulfonic acid-type (Amberlite IR 120) resin column in the acid form. The resin is washed with water and then eluted with 1 l of ammonia solution. The eluate is evaporated to dryness and the solid residue is recrystallized from isopropanol. 7.8 g (82%) of product are obtained. Melting point: 182-184 ° C.

2) S- (2-bromopropyl) penicillamine hydrobromide

The product of the previous step (7.8 g; 41 mmol) is added, while maintaining stirring, to 300 ml of a 40% acetic acid solution of hydrogen bromide. After stirring for 3 days, the reaction solution is evaporated to dryness under reduced pressure. The residue is then stirred in ether and filtered off. The solid obtained is used as such in the next step.

Weight: 14.2 g (99 Å). Melting point: 106-108 ° C.

3) 2,5-Trimethyl-1,4-thiazine-3-carboxylic acid

The above product (14.2 g; 40 mmol) is added to 500 ml of DMF containing 50 ml of triethylamine. The mixture 6 6 3 6 6 is heated for 2.5 hours in a water bath maintained at 90 ° C; it is then evaporated to dryness under reduced pressure. The residue is dissolved in 400 ml of water and the solution is filtered into 500 g of sulfonic acid resin (Amberlite IR 120) in acid form. The resin is washed with 2 L of water; then elute with 2 l of ammonia solution.

The eluate is evaporated to dryness under reduced pressure and the residue dissolved in 500 ml of water is mixed with 50 ml of carboxylic acid resin (Amberlite IRC 50) in acid form. The resin is filtered off and the aqueous phase is then evaporated to dryness under reduced pressure. The residue is dissolved in 50 ml of a 2: 1 mixture of acetone and water and the solution is placed in a condenser. After 3 days, the crystals that have appeared are filtered off. After washing with acetone and drying, 3.7 g (45%) of product are obtained so far. Melting point: 190 ° C (decomposition).

IR, NMR and mass spectra confirm the structure of the cyclized product.

Karl-Fisher analysis shows the presence of 8.7% water. Elemental analysis: C H N

% calculated 46.4 8.3 6.8% found 46.1 8.5 6.4

The product appears to be homogeneous when examined by thin layer chromatography on silica, eluting with butanol / acetic acid / water: 12: 3: 5. However, after silylation, gas phase chromatography shows the existence of two epimers in equal proportions.

Example 3. 1- (R, S) -oxo-2,2,5- (S) -trimethyl-1,4-thiazine-3- (S) -carboxylic acid

The recrystallization broth solutions of the previous example and the washing acetone of the product, which have been carefully stored, are evaporated to dryness under reduced pressure. The white solid residue is recrystallized from ethyl acetate. The solid residue 22 6 6 3 6 6 is then immediately ground in acetone and then recrystallized from a mixture of acetone and water. This gives 0.57 g (7% of 5- (2-bromopropyl) -penicillamine hydrobromide) at 98% enrichment of 2,2,5- (S) -trimethyl-1,4-

thiazine-3 (S) -carboxylic acid. Melting point: 328 ° C

20 ° (decomposition), / ot / p = 131 (C = 0.5; water).

Elemental analysis: C H N

% calculated 50.8 7.8 7.4% found 50.5 8.0 7.7

The above sulfide is dissolved in 10 ml of acetic acid and 0.4 ml of 30% hydrogen peroxide is added. The mixture is stirred for 24 hours at ambient temperature and then concentrated under reduced pressure until a syrup is obtained. 25 ml of acetone are added and the mixture is kept overnight at 5 ° C. The solid crystalline solid is filtered off and redissolved in 10 ml of water. The solution is concentrated until a syrup is obtained. The above treatment is repeated to give 0.29 g (47%) of a white solid containing a mixture of two epimeric sulfoxides. Melting point: 230 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Karl-Fisher analysis shows the presence of 2.2% water. Elemental analysis: C H N

% calculated 45.8 7.5 6.7 S found 45.6 7.4 6.9

Example 4. 2,2,5- (R) -Trimethyl-1,4-thiazine-3- (S) -carboxylic acid> .25 ml (40.3 mmol) of freshly distilled chloroacetone are added, with stirring, to a suspension of 5 g (33.6 mmol) of D-penicillamine in 28 ml of water.

After the reaction solution is homogenized, the reaction vessel is immersed in an ice bath. A 20% aqueous solution of sodium hydroxide is then added dropwise until the pH is 5.3, without allowing the temperature to exceed 10 ° C, and then a solution of 1 g (26.4 mmol) of sodium borohydride in 5 ml of water containing base, while taking care to ensure that the temperature does not rise above 30 ° C. When the addition is complete, the mixture is stirred for another hour at ambient temperature. 37% hydrochloric acid is then added slowly until the pH is 5.0. The final reaction solution is evaporated to dryness under reduced pressure and the residue is treated with 50 ml of absolute ethanol. The insoluble matter is filtered off and the filtrate is evaporated to dryness under reduced pressure. The colorless residual oil is redissolved in water and the solution is evaporated to dryness under reduced pressure. The solid obtained is mixed with 10 ml of water and the mixture is heated to reflux until the solid has dissolved. The solution is left to crystallize after 4 hours at ambient temperature and then overnight at 5 ° C. The solid is filtered off and dried. Weight: 4.3 g (20.7 mmol; 62%). Melting point: 310 ° C (decomposition). [α] D = + 93 ° (C = 1; water).

Karl-Fisher analysis shows the presence of 8.7 S of water.

IR, NMR and mass spectra are consistent with the expected structure. High performance liquid chromatography analysis shows the homogeneity of the final product.

Elemental analysis: C H N

% calculated 46.4 8.3 6.8 Ä found 46.3 8.3 6.9

When analyzed on a silica thin layer plate, the product has an Rf value of 0.69 after elution with methanol / acetic acid / chloroform: 6: 1: 4.

2 * 66366 i

Example 5. 1- (R) -oxo-2,2,5- (R) -trimethyl-1,4-thiazine-3- (S) -carboxylic acid

A solution of 3.5 g (16.7 mmol) of the product of the previous example in 17 ml of acetic acid / water: 1: 1 mixture is prepared and then cooled in an ice bath. 1.94 ml of 30% hydrogen peroxide are added dropwise over 5 minutes, maintaining stirring. When the addition is complete, the mixture is stirred for 8 hours in a cooling bath and then for 10 hours at ambient temperature. Evaporate to dryness under reduced pressure at a temperature not exceeding 60 ° C.

The residual oil is dissolved in any water and the solution is evaporated to dryness under reduced pressure. The procedure is repeated until a solid is obtained. This is then dissolved in 6 any water. 15 ml of acetone are added slowly, maintaining stirring.

The mixture is stirred for one hour at ambient temperature and filtered. This gives the first part of the desired sulfoxide.

The second portion is obtained by cooling the mother liquor to 10 ° C by adding 10 ml of acetone and then filtering the mixture after a crystallization time of 15 minutes. Total weight: 2.2 g (9.9 mmol; 59%). Melting point: 246 ° C (decomposition), [α] 25 D = -3.5 ° (C = 1; 1N HCl).

IR, NMR and mass spectra are consistent with the expected structure.

«

Analysis of the corresponding hydrochloride by the X-ray method makes it possible to confirm the absolute structure.

Kar1-Fisher analysis shows the presence of 8.2 ¾ of water. Elemental analysis: C H N

% calculated 43.0 7.7 6.3% found 43.2 7.6 6.4

When examined on a silica thin layer plate, the product has an Rf value of 0.42 after elution with methanol / acetic acid / chloroform: 6: 1: 4.

25 6 6 3 6 6

High performance liquid chromatography analysis ensures the homogeneity of the product.

Example 6. 1- (S) -oxo-2,2,5- (R) -trimethyl-1,4-thiazine-3- (S) -carboxylic acid (hydrochloride)

The crystallization broth solutions of the second part of the product of the previous example, which have been carefully stored, are evaporated to dryness under reduced pressure. The residue, weighing 1.3 g, is dissolved in 10% water and the solution is adjusted to pH 1 with 37% hydrochloric acid. The solution is concentrated to a wet weight of 3.0 g and this residue is kept at 5 ° C overnight. The solid is filtered off; it has an optical rotation in water of 79 ° C. The crystals are dissolved in any water and the solution is evaporated under reduced pressure until a syrup is obtained.

1.5 ml of acetone are added and the mixture is kept overnight at 5 ° C. Filter it. The optical rotation of the solid is 84 °.

The additional crystallization procedure produces a sufficiently pure product. Weight: 0.22 g (0.9 mmol; 5% relative to the product used in Example 4). Melting point: 275 ° C. Λ max = 85 ° (C = 1; water), + 73 ° (C = 1; 1 N HCl).

IR, NMR and mass spectra are consistent with the expected product.

Karl-Fisher analysis shows the presence of 0.5% water. Elemental analysis: C H N

% calculated 39.8 6.7 5.8% found 39.8 6.7 5.9

High performance liquid chromatography analysis shows 99.5% purity.

Example 7. 1,1-Dioxo-2,2,5- (R) -trimethyl-1,4-thiazine-3- (S) -carboxylic acid 5 g (22.4 mmol) of the product of Example 5 are dissolved in 26 6 6 3 6 6 to 300 ml of 0,5 N sulfuric acid. A solution of 2.5 g of potassium permanganate in 300 ml of water is then added over 2 hours. When the addition is complete, the mixture is stirred for 2 hours at ambient temperature and then 12 ml of formic acid are added. The mixture is filtered and then 0.5 N aqueous barium hydroxide solution is added until precipitation is complete. The mixture is filtered and the filtrate is transferred to a strong acid-type ion exchange resin column in the H + form. The resin is washed until the effluent is neutral and then eluted with 3 N ammonia solution. Concentrate the eluate under reduced pressure until a volume of about 100 ml is reached. The resulting ammonium salt solution is then filtered on a weak base type resin column. The filtrate is evaporated under reduced pressure until a syrup is obtained and 15 ml of acetone are added to the syrup. The mixture is stirred for 2 hours and then filtered. Weight: 2.7 g (11.6 mmol; 52%). Melting point: 232 ° C (decomposition), [α] D 20 = + 2 ° (C = 1; 1 N HCl).

IR, NMR and mass spectra are consistent with the expected structure.

Karl-Fisher analysis shows the presence of 6.2% water. Elemental analysis: C H N

% calculated 40.7 7.1 5.9 S found 40.7 7.3 5.9

When examined on a silica thin layer plate, the product has an Rf value of 0.38 after elution with butanol / acetic acid / water: 5: 2: 3.

Example 8. 5-Methyl-1,4-thiazine-3-carboxylic acid hydantoin 3.9 g (24.0 mmol) of 5-methyl-1,4-thiazine-3-carboxylic acid and 2.3 g (28, 8 mmol) of potassium isocyanate are added immediately successively to 25 ml of water. The mixture is heated to boiling and then the mixture is brought back to ambient temperature. The 5-methyl-4-ureido-1,4-thiazine-3-carboxylic acid solution thus obtained is then treated with 34 .mu.l of 10% hydrochloric acid. The mixture is concentrated under reduced pressure to a volume of about 20 ml. It is stirred for 1 hour at ambient temperature and then filtered. The solid is finally recrystallized from water. Weight: 2.5 g (13.4 mmol; 56 S). Melting point: 189 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: C H N

% calculated 45.2 5.4 15.0 Ä found 44.9 5.5 15.1

Example 9. 1-Ethoxycarbonylethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate (oxalate)

A solution of 3.7 g (17.4 mmol) of the product of Example 5 in 37 ml of water is treated with 0.5 N aqueous potassium hydroxide solution until the pH reaches 9. The resulting solution is evaporated to dryness under reduced pressure. The residue is suspended in 40 ml of DMF. 3.2 g (17.6 mmol) of ethyl 2-bromopropionate are then added dropwise. The reaction solution is stirred for 4B at ambient temperature, and then dissolved in 100 ml of water and extracted with chloroform. The organic extract is dried and then evaporated to dryness under reduced pressure. The residual oil is taken up in 50 ml of ethanol and then treated with a solution of 1.6 g (17.6 mmol) of oxalic acid in a minimum amount of the same solvent. The mixture is stirred for 30 minutes and filtered.

The solid thus obtained is recrystallized from ethanol.

4.3 g (10.5 mmol; 64 Å) of pure product are obtained. Melting point: 196 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

28 6 6 3 6 6

Karl-Fisher analysis shows the presence of 3.1% water. Elemental analysis: C H N

% calculated 44.1 6.5 3.4% found 44.1 6.7 3.3

When chromatographed on a silica thin layer plate, the product has an Rf value of 0.52 after elution with butanol / acetic acid / water: 5: 2: 3.

Example 10. 1-Pivaloyloxyethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate (oxalate)

A solution of 3.7 g (16.4 mmol) of the product of Example 5 in 37 ml of water is treated with 0.5 N aqueous potassium hydroxide solution until the pH reaches 9. The resulting solution is evaporated to dryness under reduced pressure. The residue is suspended in 40 ml of DMF. 2.9 g (17.6 mmol) of 1-chloroethyl pivalate are then added dropwise with good stirring. After the addition is complete, stirring is continued for 48 hours at 40 ° C. The resulting reaction solution is diluted with 100 ml of water and then extracted with chloroform.

The organic extract is dried and then evaporated to dryness under reduced pressure. The residue is treated with 50 ml of ethanol and then with a solution of 1.6 g (17.6 mmol) of oxalic acid in a minimum amount of the same solvent. The mixture is stirred for 30 minutes and then.

the precipitate that has formed is filtered off. It is finally purified by recrystallization from ethanol. This gives 4.2 g (9.9 mmol; 60 S) of a white solid crystalline solid. Melting point: 214 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: C H N

% calculated 48.2 6.9 3.3% found 48.1 7.2 3.2 25 66366

When chromatographed on a silica thin layer plate, the product has an Rf value of 0.65 after elution with butanol / acetic acid / water: 5: 2: 3.

Example 11. Methyl 2,2,5-trimethyl-1,4-thiazine-3-carboxylate (hydrochloride)

A solution of 32.2 g of potassium hydroxide in 640 ml of methanol is cooled to 0-5 ° C. 47.9 g (0.24 mol) of penicillamine methyl ester hydrochloride are added, followed by dropwise addition of 25.0 g of chloroacetone. The reaction solution is stirred for 1 hour at an initial temperature and then adjusted to pH 6 x 5 with ethanolic hydrogen chloride solution. 6.2 g of sodium borohydride are then added in small portions. When the addition is complete, the mixture is stirred for a further 1 hour at the same temperature. Excess hydride is destroyed by the addition of concentrated hydrochloric acid and then the solution is neutralized with 10% aqueous bicarbonate solution. The aqueous solution is extracted with chloroform. The extracts are dried and then evaporated to dryness under reduced pressure. The residual oil is dissolved in a mixture of 50 ml of methanol and 350 ml of ether. Addition of saturated hydrogen chloride ether solution causes precipitation of the solid hydrochloride. This is filtered off and reprecipitated twice from the methanol solution by adding ether. Weight: 17.2 g (30 Å). Melting point: 204 ° C (decomposition).

TR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: C H N

% calculated 45.1 7.6 5.8% found 45.1 7.7 5.8

Example 12. 2,2,4,5-Tetramethyl-1,4-thiazine-3-carboxylic acid 1.68 g (20 mmol) of sodium bicarbonate and 1.5 g (0.66 ml; 10.5 mmol) of methyl iodide is immediately added successively to a solution of 2.1 g (10 mmol) of 2,2,5-trimethyl-1,4-thiazine-3-carboxylic acid in 50 ml of methanol. The resulting reaction solution is stirred at ambient temperature for 24 hours and then heated at reflux for the same time. The solution is evaporated to dryness under reduced pressure and the residue is dissolved in 100 ml of water. The solution thus obtained is transferred to a strong acid-type ion exchange resin column in the H + form. The resin is washed with water until the effluent is neutral and then eluted with 1 N ammonia solution. The eluate is evaporated to dryness under reduced pressure. The white solid residue is recrystallized from ethyl acetate. 1.4 g (6.8 mmol; 68%) of pure product are obtained. Melting point: 201 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: 'C H N

% calculated 53.2 8.4 6.9% found 52.8 8.5 7.0

When chromatographed on a silica thin layer plate, the product has an Rf value of 0.60 «» after elution with ethanol.

I:

Example 13. 5-n-Butyl-2,2-dimethyl-1,4-thiazine-3-carboxylic acid (hydrochloride) 7.5 g (50 mmol) of D-penicillamine are added to any water maintained at 0 ° C. C to 5 ° C. At the same time, dropwise, while maintaining good stirring, add the following ingredients: other side of 9.5 g (53 mmol) of L-bromohexan-2-one in about 10 minutes and the other half with 25 ml (50 mmol) of 2 N aqueous sodium hydroxide solution for about 30 minutes during the . When the addition is complete, stirring and temperature are maintained for the next one hour and then the temperature is adjusted to about 15 ° C. 50 ml of methanol are added, followed slowly by a solution of 1.5 g (39 mmol) of sodium borohydride in 9 ml of water containing the base. The reaction solution is left to return to 31,636,366 ambient temperature and then extracted with chloroform. The organic extract is dried and then evaporated to dryness under reduced pressure. The red residual oil is dissolved in 50 diisopropyl ether and sufficient ethanol is added to give a solution. An ethereal solution of hydrogen chloride is then added until precipitation is complete, followed by sufficient ethanol to give a light gray-brown solid from the tan mass.

This is filtered off, washed with diisopropyl ether and dissolved in hot ethanol. Ethyl ether is added until turbidity remains and then the mixture is allowed to cool. The crystals are collected and purified a second time. Weight: 8.0 g (30 mmol; 60 S »). Melting point: 225 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: C H N

% calculated 49.3 8.3 5.2% found 49.5 8.2 5.5

When chromatographed on a silica thin layer plate, the product has an Rf value of 0.62 after elution with butanol / acetic acid / water: 5: 2: 3. J

i

Example 14. 5-Methyl-2-phenyl-1,4-thiazine-3-carboxylic acid (hydrochloride) 1 g (4.3 mmol) of threo-6-phenylcysteine hydrochloride is dissolved in 12 ml of water. The solution is cooled in an ice bath and then the pH is adjusted to 7.0 with 20% aqueous sodium hydroxide solution. 0.42 g (4.5 mmol) of freshly distilled chloroacetone are added. After the pH has stabilized for 30 minutes, 2N aqueous sodium hydroxide solution is added until neutralization is achieved. The mixture is stirred for a further 1 hour, still in an ice bath and then a solution of 0.13 g (3.4 mmol) of sodium borohydride in 2 ml of water containing base is added. A clear, colorless solution with a pH of 9.1 is obtained. The solution is stirred for 1 hour, allowing 32 66366 to return to ambient temperature. The pH is adjusted to 6.0 with 37% hydrochloric acid. The mixture is evaporated to dryness under reduced pressure. The residue, suitably dried, is dissolved in 10 ml of ethanol and 2 ml of a 22% ethanolic hydrogen chloride solution are added. The mixture is stirred for 15 minutes and then evaporated to dryness under reduced pressure. The residue is recrystallized twice from isopropanol / ethanol: 4: 1. Weight: 0.66 g (2.4 mmol} 56 55). Melting point: 248 ° C (decomposition).

IR, NMR and mass spectra are consistent with the expected structure.

Elemental analysis: C H N

Ä calculated 52.6 5.9 5.1% found 52.3 5.9 5.4

Example 15. 1- (R) -oxo-2,2,5- (R) -trimethyl-1,4-thiazine-3- (S) -carboxylic acid (without isolation of intermediate) 7.4 g (80 mmol) of root distilled chloroacetone is added, maintaining good stirring, to a suspension of 10 g (67 mmol) of (D) -penicillamyr in 56 ml of water at ambient temperature. After a reaction time of 2 hours, 20% aqueous sodium hydroxide solution is added until the pH is 5.3 (temperature <20 ° C). A solution of 2 g (50 mmol) of sodium borohydride in 10 ml of water is added dropwise to the reaction solution (temperature έ 30 ° C). After a reaction time of 1 hour, the solution is acidified by adding 50 ml of acetic acid. The reaction solution is cooled (0 ° C ύ temperature 65 ° C) and 2.4 g (0.74 mol) of 30% hydrogen peroxide are added dropwise. The temperature is maintained between 0 ° C and 5 ° C for 8 hours and then at 20 ° C for 4 hours. The mixture is concentrated under reduced pressure. The residue is dissolved in 40 ml of ethanol, the suspension is filtered and the ethanolic solution is added to 240 ml of ethyl acetate, maintaining good stirring. The product is left to crystallize at 20 ° C for 1 hour. The precipitate is filtered off and washed with acetone. The crude amino acid is crystallized from H 2 O / ac 33: 36366: 1: 4. 7.4 g (33.3 mmol; 54%) of a white crystalline substance are obtained, which is identical to the product of Example 5.

Table I below lists the derivatives of the above examples along with other derivatives of the invention prepared according to the above methods.

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k 41 66366 (1): This is most often the Decomposition temperature. The value entered in the table corresponds to the initial value of the phenomenon.

(2): The correct C, H, N elemental analysis is obtained for all the products mentioned (3): Hydrated product (4): Hydrochloride (5): Free base or zwitterion (6): Oxalate (7): Mixture of isomers, the separated components of the mixture are shown in Table (8) below: Configuration of Asymmetric Centers: 1- (R) -3- (S) -5- (R) (9): Configuration of Asymmetric Centers: 1- (S) -3- (S) -5 - (R) (10): Configuration of asymmetric centers: 1- (R) -3- (S) -5- (R) (11): Configuration of asymmetric centers: 3- (S) -5- (S) (12): Configuration of centers of asymmetry: 1- (R, S) -3- (5) -5- (S) (13): phth denotes 3-phthalidyl (14): pyr denotes 2,5-dioxo-1-pyrrolidinyl (15) : Configuration of Asymmetric Centers: 3- (R) -5- (S) (16): Configuration of Asymmetric Centers: 1- (S) -3- (R) -5- (S) (17): Configuration of Asymmetric Centers: 3- (S) ) -5- (R) (18): Hemihydrochloride

The products of the invention were subjected to a series of pharmacological tests, the methodology of which is described below. The results of these tests are recorded in Table II, where the numbers in the first column correspond to the numbers in the first column of Table I.

LD50 values are calculated according to the method of Lichtfield and Wilcoxon (J. Pharmacol. Pxp. Ther. 96, 99, 1949) and are expressed in mg / kg. The products have been administered orally to mice.

The effect of the substances has been studied using the method derived from the method of S. Irwin (Gordon Res. Conf. On Medicinal Chem., 133, 1959). The compounds suspended in mucilage containing 1% tragacanth gum are administered orally by gavage to groups of 5 male mice (CD1 strain, Charles River, fasted for 18 hours).

Doses tested as a function of observed activity range from 3000 to 3 mg / kg. The effect has been studied at 2, 4, 6 and 24 hours after treatment. Observation has been continued if the effect continues at this stage. Mortality was recorded within 14 days of treatment. None of the products tested caused abnormal behavior in mice or were shown to be toxic.

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Determination of the effect on fibrinolytic activity, bleeding time and blood loss

The test compound, suspended in mucilage containing 1% gum tragacanth, is administered orally to rats at a dose of 30 mg / kg. A blood sample is taken 1 hour or 3 hours after treatment. Plasma is separated by centrifugation and euglobulins are precipitated. Fibrinolytic activity is determined by the method of Astrup and Mullertz (Arch. Biochem. Biophys. 1952, 4Ό, 346). 50 of a solution of eugolobulins in veronal buffer containing 0.25% gelatin is placed on plates with 0.08% fibrin suspension. After an incubation time of 18 hours at 37 ° C, the diameter of the room zone is measured.

The score is a function of the ratio of the fibrinolytic activity of the treated rats to the fibrinolytic activity of the placebo-treated rats. The meanings are as follows:

Score 4: Growth constant of fibrinolytic activity 0.51 - 0.99

Score 5: Growth constant of fibrinolytic activity 1 to 1.5

Score 6: Growth constant of fibrinolytic activity: C

1.6 -2

Score 7: Fibrinolytic activity growth constant 2.1 to 2.5

Score 8: Growth rate of fibrobinolytic activity 2.6 to 3

Score 9: Growth constant of fibrinolytic activity £ 3.1

To determine the bleeding time, the tail of the rats, kept at ambient temperature for 2 hours, is cleaned with ether and the end of the tail is incised with a knife 2 mm. The leakage time, expressed in seconds, is measured 1 hour or 3 hours after treatment. The blood is collected on filter paper. The amount of blood loss i 48 66366 is determined by weighing the filter paper before and after blood collection.

The growth constant for bleeding time is the ratio of the bleeding time of treated rats to the bleeding time of placebo-treated rats.

Point value 3 means growth constant 0.79 - 0f89 Point value 4 means growth constant 0.9 - 1.09 Point value 5 means growth constant 1.1 - 1.11 Point value 6 means growth constant 1.12 - 1.22

A score of 7 denotes a growth constant of 1.23 to 1.33

A score of 8 denotes a growth constant of 1.34 to 1.44

A score of 9 denotes a growth constant of 2 1.45

The growth constant of blood loss is the ratio of the amount of blood lost in treated rats to the amount of blood lost in control rats.

Point value 3 means growth constant έ 0.5 Point value 4 means growth constant 0.51 - 0.99 Point value 5 means growth constant 1 - 1.5 Point value 6 means growth constant 1.6 - 2

A score of 7 denotes a growth constant of 2.1 to 2.5

A score of 8 denotes a growth constant of 2.6 to 3

A score of 9 denotes a growth constant of 2 3.1

The two products of the invention were the subject of a more detailed evaluation: These are derivative 7 and derivative 12 (free base of derivative 7 in hydrated form).

Compound 7 was the subject of a one-month toxicity study in rats and monkeys. This study showed that it is well tolerated up to a dose of 300 mg / kg / day.

The toxicity of compound 12 is excellent: LD50 g is greater than 8 g / kg in mice and rats. In monkeys, it is well tolerated up to a dose of 800 mg / kg.

49 66366

When derivative 7 is administered to 4 groups of rabbits at doses of 2.5 to 50 mg / kg, an increase in blood fibrinolytic activity is observed that is proportional to the dose administered. For example, this increase is 3-fold 90 minutes after dosing of 5 mg / kg. This effect is evident at continuous dosing; thus, treatment at a dose of 10 mg / kg twice daily for 21 days increases blood fibrinolytic activity to 3 times that of control animals.

When the derivative 12 is administered intravenously at doses of 3 or 5 mg / kg / 10 minutes for two hours, a 2.8-fold increase in fibrinolytic activity is observed from 10 to 30 minutes after the start of administration for at least 3 hours until the end of administration.

The activity of Compound 12 was also demonstrated in rats and monkeys when administered orally. Finally, in humans, an oral dose of 100 to 800 mg causes an increase in fibrinolytic activity that is dose proportional.

When the novel compounds of the invention are administered, the daily dose is 50 mg to 2 g when administered orally and 10 mg to 1 g when administered by injection or intravenous infusion.

The products of the invention can be used in various galenic forms. The following examples are not intended to be limiting, but relate to galenic formulations containing the active product of the invention, indicated by the letter A, which may be, for example, 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylic acid, ethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 2,2,4,5-tetramethyl-1,4-thiazine-3-carboxylic acid, 1- (pivaloyloxy) ethyl-1 -oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, (octyloxy-ceronyl) methyl-1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 1- (ethoxycarbonyl) ethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 5-butyl-2,2-dimethyl-1,4-thiazine-3-carboxylic acid, 2 , 2-dimethyl-50,663,665 5-isopropyl-1,4-thiazine-3-carboxylic acid, 5-methyl-Z-phenyl-1,4-thiazine-3-carboxylic acid or ethyl-3-carboxy-2 , 2-dimethyl-l, 4-thiazine-5-acetate.

Coated pills A 50 mg

Sta RX starch 100 mg

Microcrystalline cellulose 60 mg

Extra fine lactose, crystals 132 mg

Colloidal silica 2 mg

Hydrogenated castor oil 6 mg A 100 mg

Dicalcium phosphate 95 mg

Sodium glycolate starch 52.5 mg

Colloidal silica 2 mg

Magnesium stearate 0.5 mg

Tablet A 500 mg

Mannitol 95 mg

Polyvinylpyrrolidone 27 mg

Polyethylene glycol 6000, powder 8 mg A 300 mg Starch 60 mg

Lactose 130 mg

Low viscosity ethylcellulose 20 mg

Talc 5 mg

Aerosil 5 mg Suppositories A 500 mg α-tocopherol 10 mg

Witepsol H 15 (semi-glyceride) 2600 mg 5i 6 6 3 6 6

Solutions for injection A 20 mg

Sodium chloride 2.5 mg

Citric acid / sodium citrate buffer to pH 4.8 A 50 mg

Propylene glycol 300 mg

Ethyl alcohol 50 mg

Water for injections, volume 1 ml Coated pill containing 100 mg dose of Compound A and 300 mg dose of acetylsalicylic acid A 100 mg

Acetylsalicylic acid 300 mg

Ethylcellulose 60 mg

Lactose 40 mg

Aerosil 8 mg

Magnesium stearate 8 mg

Tablets containing 300 mg of Compound A

and a 50 mg dose of dipyridamole A 300 mg

Dipyridamole 50 mg

Hypromellose 25 mg Starch 100 mg

Calcium stearate 8 mg

A tablet containing a 200 mg dose of Compound A and a 100 mg dose of Suloctidil A 200 mg

Suloctide 100 mg

Tvi / een 80 8 mg

Ethylcellulose 22 mg

Lactose 40 mg Starch 71 mg

Magnesium stearate 9 mg

Claims (8)

A process for the preparation of 1,4-thiazine-3-carboxylic acid derivatives of the formula I (O) R 'n 1 R 7 / S 2 (I) π N ^ Cl 1 »^ r5 r4 0 in which R R 2 and R 2 ', which may be the same or different, denote hydrogen, a straight or branched C 1-4, C 2-, Cy or C 4 alkyl radical or phenyl optionally substituted by a halogen atom, such as fluorine, chlorine or bromine, straight or branched With a C 1-4, C 1-4, C 3-4 C 1-4 alkyl radical, or a straight or branched C 1-, C 2-, C 3- or C 4- alkoxy radical, R 1 and R 2 may form together with an adjacent carbon atom a cycloalkyl having 3, 4 , 5, 6, 7 or 8 carbon atoms, where one or more carbon atoms may be optionally replaced by a sulfur or oxygen atom or by the group SO, SO2 or NRg, where Rg represents hydrogen, straight or branched C1-4, c2-C3 * (C4 alkyl radical, phenyl or benzyl group, R3 denotes; - hydroxyl group, - group ORg, in which Rg is represents a straight or branched ^ 1 "'C2"' ^ 3 "r ^ 4" r C5 "'C6"' C7 "r c8" r ^ 9 "» ^ 10 "r Cl1" or a C1-12 alkyl group, a group O-CH -COORi 1 or a group I XX R 10 OCH-OCOR ^, in which R ^ q represents hydrogen or Cy or C 2-4 alkyl-R 10 radical and R j ^ represents a straight or branched C 1-6. 3 ”'^ k ~' ^ 5” »^ 6” »^ 7" »C3_» ^ g ~ »C1Q-, C ^ - or C ^ -3]. - alkyl radical, '^ 12 - group Νχ, where R ^ And ^ 33 *, which may be the same or different, denote hydrogen or a straight or branched C1-, C2-> C3- or C1-4-alkyl radical, or - a 3-phthalidyloxy group or a 1- (2,5-dioxopyrrolidinyl) - 1-ethoxy group, R 1 denotes a hydrogen atom, a straight or branched C 1-4, C 3-, C 3 "C 4" * C 5 "" C 6 ~ * C 7 "or a C 6 -C 6 alkyl radical, a straight or branched C A, C 1-4, C 1-4, C 1-4, C 1-4 or C 1-4 acyl group or a group CON 1 · ^ 3, R 3 and R 4 may form a hydantoin ring with adjacent carbon and nitrogen atoms, R 5 R 6 'and may be the same or different, denote hydrogen , a straight or branched C 1-6, C 2-4 C 1-4 alkyl, optionally substituted by a halogen atom such as fluorine, chlorine or bromine, a straight or branched C 1-4, C 2-4 alkyl or C 1-4 alkyl radical or a straight or branched C 1-4, C 1-4 or C 1-4 alkyl radical or a straight or branched C 1-4, C 2-, C 1-6 alkyl or With a C 1-4 alkoxy radical, or a group CH 2 CO 4, in which R 1 represents hydrogen or a straight or branched C 1-4, C 1 -C 4 or C 1-4 alkyl radical, denotes hydrogen, a straight or branched C 1-4, C 2, C a 4- or C 1-4 alkyl radical, a geminal dimethyl radical or phenyl optionally substituted by a halogen atom, such as fluorine, chlorine or bromine, a straight or branched C 1-4, C 2-, C 1-4 or C 1-4 alkyl radical or a straight or branched C 1-4 alkyl radical, -, C 1-4, C 1-4 or C 1-4 alkoxy, and n may have the value 0, 1 or 2; if R 1, R 2, R 4 and R 7 represent hydrogen and if R? represents a hydroxyl group or a straight or branched C 1 -C 4 alkyl radical, R 1. or R 1 does not represent hydrogen, a straight or branched C 1 -C 4 alkyl radical or phenyl, and also for the preparation of salts of these compounds with pharmaceutically acceptable acids and 54 6 636 6 bases, characterized in that: a) a derivative of formula II? ^ C0R3 R6-C-CH / nh-r, / 5 h is cyclized in a basic reaction solution, or b) a derivative of formula III R, ^ COR, Ro-C-CH / \ III (O) CH C - Rt II o R 7 R 6 wherein X represents a halogen atom such as chlorine, bromine or iodine, a hydroxyl group or a group which can be easily removed, such as a tosyl or mesyl group, is cyclized in a basic or dehydrating reaction solution, or c) 5.6 -dihydro-2H-thiazine derivative VI R7s / R2 VI Rs "^ N / AcOR3 55 66366 is reduced to 1,4-thiazine of formula I, optionally followed by oxidation to give the corresponding sulfoxide or sulfone, or d) in the preparation of a derivative of formula I wherein haarau a known C 1 -C 4 alkyl radical or a straight or branched C 1 -C 4 acyl radical, the amine of formula I containing hydrogen is alkylated or acylated in a basic reaction solution with a compound R 1 Z in which Z represents a halogen atom such as chlorine, bromine or iodine, or a group which can be easily removed, such as, for example, a tosyl or mesyl group, or e) in the preparation of a derivative of formula I in which R 1 represents a radical CONH 2, a derivative of formula I in which R 1 represents hydrogen is reacted with a compound having an MNCO of formula wherein M represents a monovalent cation such as sodium, potassium, lithium or ammonium ion, or f) in the preparation of a derivative of formula I wherein R 1 and R 2 form a hydantoin ring with adjacent carbon and nitrogen atoms, a derivative of formula I wherein R 1 ^ represents a hydroxyl group and R ^ represents a group CON1 · ^, is brought into an acidic reaction solution by heating, if necessary, or g) in the preparation of a derivative I in which n = 1 or 2, of the formula A derivative of I in which n = 0 is treated with an oxidizing agent, or h) a derivative of formula I in which R 1 represents a hydroxyl group is converted into a derivative in which OR 1, OCHCOOR 3 or OCHOCOR 3 by esterification, or R 10 R 10 i) a derivative of formula I, wherein R 1 represents a hydroxyl group or a group OR 2, OCHCOOR 4 or OCHOCOR 4, is converted to a derivative-R 10 to R 10 to a derivative in which R 2 represents a group N "R 12 * by amidation, or R 13 j) a derivative of formula I in which R 1 represents an group OR 1, OCHCOOR 3 or OCHOCOR 4 is converted to a derivative in which R 10 R 10 R 1 represents a hydroxyl group or a group N-R 2 -hydrolyzed by R13 or aminolysis, or k) a derivative of formula I in which a group N-R 2 -R 13 is converted to a derivative in which R 1 represents a hydroxyl group or a group OR 2, OCHCOOR 4 or OCHOCOR 3, by hydrolysis of R 10 to R 10 or by alcoholization. Process according to Claim 1, characterized in that in the general formula I and R 1, which may be identical or different, represent hydrogen or a methyl group. Process according to Claim 1 or 2, characterized in that in the general formula IR 1 represents a hydroxyl group, a group OR 1 in which R 2 represents a straight or branched C 1 -C 4 alkyl radical, a group OCH-COOR 2 R 10 or a group OCHOCOR 3 in which R 1g represents hydrogen or methyl and the radical R 1g represents a straight or branched C 1 -C 6 alkyl radical. Process according to any one of Claims 1 to 3, characterized in that, in the general formula, I-R 2 denote hydrogen or a methyl radical. Method according to any one of claims 1 to 4, characterized in that in the general formula I R 1. and R, which may be the same or different, denote hydrogen, a straight or branched C 1 -C 4 alkyl group or a group CH 2 COOR 4 in which R 1 is a hydrogen or C 1 -C 4 alkyl group. 5 ?. 6 6 3 6 6 Process according to Claim 5, characterized in that, in the general formula I, R 1 represents hydrogen and R 4 represents a straight-chain or branched C 1 -C 4 alkyl group. Method according to one of Claims 1 to 6, characterized in that in the general formula I n has the value 0 or 1. Process according to Claim 1, characterized in that the compound to be prepared is selected from the group consisting of the following compounds: 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylic acid, ethyl-1-oxo -2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 2,2,4,5-tetramethyl-1,4-thiazine-3-carboxylic acid, 1- (pivaloyloxy) ethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, (octyloxycarbonyl) methyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 1 - (ethoxycarbonyl) ethyl 1-oxo-2,2,5-trimethyl-1,4-thiazine-3-carboxylate, 5-butyl-2,2-dimethyl-1,4-thiazine-3-carboxylic acid, 2,2- dimethyl-5-isopropyl-1,4-thiazine-3-carboxylic acid, 5-methyl-2-phenyl-1,4-thiazine-3-carboxylic acid and ethyl-3-carboxy-2,2-dimethyl-1,4-thiazine- 5-acetate. 58 6 6 3 6 6