SK39097A3 - Bicyclic amidine derivatives useful in therapy - Google Patents

Abstract

Compounds of the formula (I) wherein D represents a 5-membered heterocyclic aromatic ring containing 1 to 4 heteroatoms selected from O, N or S, optionally substituted at a carbon atom by halogen, trifluoromethyl, alkyl C1 to 6, nitro, cyano, and which is connected to the remainder of the compound of formula (I) through a carbon atom; A represents (N(X) or CH(-(CH2)m-NXY); U represents NH, O or CH2; V represents (CH2)b; a, b, m, X and Y are as defined in the specification, together with processes for their preparation and compositions containing them. Compounds of formula (I) are nitric oxide synthetase inhibitors and are useful in therapy.

Description

Bicyclic amidine derivatives useful in therapy

Technical field

The present invention relates to bicyclic amidine derivatives, processes for their preparation, compounds containing them and their use in therapy.

BACKGROUND OF THE INVENTION

Some amidine derivatives have been reported to be useful in therapy. According to US No. 3669974 (USV Pharmaceutical Corp.) of UK Patent Application 2226562 uses N-phenylamidine derivatives for the treatment of diabetes. International Patent Application WO 92/04054 (University of Oregon) discloses the use of N ', N' '- disubstituted admidines in the treatment of hypertension, depression and hallucinogenic conditions. The use of some amidines and symmetrical bisamidines as a US patent and Boots has been described

analgesics, in treatment inflammation and in treatment hypertension is described in Belgium Patent No. 717740 a in the UK patent No. 1180629 (both patents from Delalande). In German application Invention German Patent application DE-OS-2321330

(Bayer) discloses the use of amidine derivatives as herbicides.

Also described is the use of nitric oxide synthetase (NO) synthase inhibitors in the treatment of diseases, e.g. in International Patent Applications WO 94/12163 (Abbott), WO 93/13066 and WO 94/12165 (both Wellcome) and European Patent Application 446699 (Merrell Dow), 547558 and 558468 (both Washington University). The use of nitric oxide synthease inhibitors in therapy is also described in WO 95/00505, WO 95/09619, WO 95/09621 (all Wellcome), WO 95/10266 (Otsuka), WO 95/11231 and WO 95 / 11014 (both Searle), six of which were published after the earliest priority date you give to this application.

The Applicant has previously described the use of guanidine derivatives and amidine derivatives which are inhibitors of NO synthase in the treatment of, inter alia, neurodegenerative diseases (WO 94/21621, WO 95/05363). The second of these patents was published after the earliest possible priority date of this application.

SUMMARY OF THE INVENTION

We have now found a new class of bicyclic amidine derivatives having useful pharmaceutical activity.

According to a first aspect of the present invention, we have prepared a compound of formula I

where

D represents a five membered heterocyclic aromatic ring containing from 1 to 4 heteroatoms selected from O, N or S, optionally substituted on a carbon atom with halogen, trifluoromethyl, C 1-6 alkyl, nitro or cyano, and which is attached to the remainder of the compound of formula I via a carbon atom;

A is N (X) or CH (-CH ₂ ) _m -NXY);

U represents NH, O or _CH2;

W represents (CH _{2) a;}

W represents (CH ₂ ) _b ;

a and b independently represent integers from 0 to 3, provided that a + b is in the range of 1 to 3;

X and Y independently represent hydrogen, alkyl C1 to 6, or - (CH _{2) n} Q, or NXY represents piperidinyl, pyrrolidinyl, morpholinyl or tetrahydroisoquinolinyl;

Q is biphenyl or phenyl optionally substituted with one or more groups selected from C1-6 alkyl, C1-6 alkoxy, C1-6 perfluoroalkyl, halogen, nitro- or cyano-;

m is an integer from 0 to 5;

n is an integer from 0 to 6;

or the chain U-V-A-W is defined as above but may be unsaturated;

or the UVAW chain may be represented by an -NH-CH ₂ -CH ₂ -O- chain substituted on a carbon atom by a - (CH ₂ ) _m -NXY group, wherein m, X and Y are as defined above, and the corresponding pharmaceutically acceptable salts.

A preferred group of compounds of formula I is defined by formula IA

NH

NH where

T is a C ₃ _ ₅ saturated or unsaturated alkylene chain substituted with ~ _{(CH2) m} -NXY; -O (CH ₂ ) ₂ -NHsubstituted - (CH ₂ ) _m -NXY; or -U- (CH ₂ ) _a - N (X) - (CH ₂ ) _b -;

X and Y are each independently hydrogen, C 1-6 alkyl, or - (ΟΗ ₂ ) 4 O 'or -NXY is piperidinyl, pyrrolidinyl, morpholinyl or tetrahydroisoquinolinyl;

Q is phenyl optionally substituted with C1-6 alkyl, C1-6 alkoxy, trifluoromethyl, halogen, nitro or cyano; and

U, m, n, a, b and D are as defined above, but provided that when T is -U- (CH ₂ ) _and

-N (X) - (CH ₂ ) and X represents - (CH ₂ ) _n Q, represents an integer between 0 and 5, and the corresponding pharmaceutically acceptable salts.

Preferably, D represents a five membered heterocyclic aromatic ring containing one heteroatom selected from O, N or S, optionally substituted on a carbon atom by halogen. It is particularly preferred that D is thienyl, furyl or pyrrolyl, especially thienyl or furyl, especially thienyl and most preferably 2-thienyl.

We prefer that T represents C ₃ _ ₅ saturated or unsaturated alkylene chain substituted by - (CH _{2) m} -NXY, particularly a saturated ^C 3-5 alkylene chain substituted by - (CH2) m-NXY, especially C3_4 saturated alkylene chain substituted by - ^(CH 2) _m -NXY.

When T represents a C ₃ _ ₅ saturated or unsaturated alkylene chain substituted by - _{(CH2) m-} NXY; or -O- _{(CH2) 2-NHsubstituovaný} - (CH _{2) and} NXY, we prefer that x and y each independently representing H, alkyl, Cl particularly prefer that X is hydrogen, methyl, ethyl or a group, either X or Y is H eq H or - (CH _{2) n} Q.

to 6 or - (CH ₂ ) _n Q.

. and Y independently represents

- (CH ₂ ) _n Q ^and in particular that the symbol and the second symbol in order to represent m prefer to be 0 or 1, in particular 0.

If T represents -U- (CH ₂ ) _and ^_ N (X) - (CH 2 -) -, we prefer U to represent CH ₂ .

If T is -U- (CH ₂ ) _and -N (X) - (CH ₂ ) _b -, we prefer that a + b be 1 or 2.

When T represents -U- _{(CH2) &} -Ν (Χ) - (0Η ₂₎ ^ -, we prefer that X representing hydrogen, alkyl Cl to 6 or the group

- (CH ₂ ) _n Q.

If X and / or Y is - (CH ₂ ) _n Q, we prefer that n be 0, 1 or 2, especially 1.

We prefer Q to represent phenyl optionally substituted by C5 alkyl or halogen, although it is particularly preferred that Q is unsubstituted phenyl.

According to the present invention, compounds of the general formula of salts comprising:

further protected is the process for the preparation of I and their pharmaceutically acceptable (a) preparation of the corresponding compound of formula II

I reaction

NH ₂ II wherein, U, V, A and W correspond to the symbols defined above, with a compound of formula III

NH / \

D

III wherein D is as defined above and L is a leaving group;

(b) preparing a compound of the general formula IV corresponding to formula I by reaction

IN

I

A.

.For.

W

H-NH ₂ -HA

Wherein U, V, A and W are as defined above and HA is an acid, with a compound of formula V

Where D is defined as above;

(c) a compound of formula I wherein A represents N (X) and X represents alkyl C1 to 6 or - (CH _{2) n} Q by reacting a corresponding compound of formula I wherein X is hydrogen with a compound of formula VI

R ⁹ -L VI wherein R ⁹ is C 1-6 alkyl or - (CH ₂ ) _n Q and L is a leaving group;

(d) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY) and at least one X or Y is C 1-6 alkyl or - (CH ₂ ) _n Q, by reacting the corresponding compound of formula Wherein one or both of X and Y are hydrogen with a compound of Formula VI;

(e) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY) and _m is an integer from 1 to 5, by reducing the corresponding compound of formula VII

XYNCO (CH ₂ ) mi-

VII wherein U, V, W, X, Y and D are as defined above;

(f) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY) and both X and Y are hydrogen, by reducing the corresponding compound of formula VIII

O ₂ N- (CH ₂ ) m

VIII wherein U, V, W, m and D are as defined above;

(g) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY), X is hydrogen and _m is an integer from 1 to 5, by reducing the corresponding compound of formula IX

YN = CH (CH ₂ ) _m -I-

NH

IX wherein U, V, W, D and Y are as defined above;

(h) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY), X or Y is hydrogen and the other is - (CH ₂ ) _n Q wherein n is an integer from 1 to 6, by reducing the corresponding compound of formula X, II-4-NH

Q (CH ₂ ) _{n -} CONH (CH ₂ ) n - ^H - = NH X

D wherein Q, m, U, V, W and D are as defined above;

(i) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY), X or Y is hydrogen and the other is - (CH ₂ ) _n Q wherein n is an integer from 1 to 6, by reducing the corresponding compound of formula XI

Q (CH ₂ ) _n .CH = N- (CH ₂ ) m -

XI wherein Q, m, U, V, W and D are as defined above; or (j) preparing a compound of formula I wherein A is CH (-NXY) and X is hydrogen, by reducing the corresponding compound of formula XII

wherein U, V, W, D and Y are as defined above;

and if desired or necessary converting the resulting compound of formula I or another salt thereof into a pharmaceutically acceptable salt, or vice versa.

In process (a), the reaction is carried out while stirring the mixture of reactants in a suitable solvent, e.g. in a lower alkanol such as ethanol, isopropanol or tertiary butanol at a temperature between room temperature and the reflux temperature of the solvent used. The reaction time depends inter alia on the solvent and the nature of the leaving group and may be up to 48 hours, but typically is from 1 to 5 hours. The corresponding leaving group L may be thioalkyl, sulfonyl, trifluoromethylsulfonyl (trifluorocarbon sulphonyl), halide, alkyl and aryl alcohols and tosyl; other groups are described in Advanced Organic Chemistry, J.March (1985), third edition, McGraw-Hill on p. 315; these groups are well known in the art.

In process (b), the reaction is preferably carried out by refluxing a mixture of the two compounds for several hours in the presence of a suitable solvent, the reaction temperature being high enough to condense the solvent in the reflux condenser but not so high as to cause decomposition of the formed amidine. The reaction temperature may range from room temperature to about 250 ° C, but is preferably carried out at temperatures between about 100 ° C and 200 ° C. It has been found that o-dichlorobenzene is a particularly good solvent and that it is preferable to add 4-dimethylaminopyridine as a catalyst. Upon cooling, two layers are formed, the solvent can be decanted and the reaction mixture treated after addition of aqueous base.

In those cases where the reactants are soluble in the solvent, the solvent can be evaporated under reduced pressure and the reaction mixture treated with water. HA can be organic or inorganic hydrochloric, hydrobromic, phosphoric, acetic, lactic, acid, for example hydroiodic, sulfuric, nitric, succinic, fumaric, malic, maleic, tartaric, citric, benzoic or methanesulfonic acid.

In process (c), the reaction is carried out under conventional conditions, for example by reacting the two compounds in an inert solvent under basic conditions at room temperature for up to 12 hours. It has often been found that it was convenient to treat the amine with NaH before reacting with the compound of formula VI. Preferably, L is a halide, especially bromide.

Process (d) may be carried out under the conditions described in process (c).

In process (e), the reduction can be carried out by treatment with diborane in an inert solvent such as THF. Alternatively, although less preferred agents may include lithium aluminum hydride and catalytic hydrogenation agents, for example H ₂ to Pd / C. Further details of the reaction conditions that can be used in these reactions can be found in J. March: Advanced Organic Chemistry, p. 1099 including references cited herein.

In process (f), the reduction can be carried out under various reaction conditions, for example those described in J. March: Advanced Organic Chemistry on p. 1103-1104. These conditions include catalytic hydrogenation, use of metallic Zn, Sn or Fe, AlH-β-AlClg, sulfides and others. We prefer to carry out this reaction by hydrogenation at atmospheric pressure for 3 to 6 hours in the presence of a palladium-carbon catalyst.

In processes (g), (i) and (j), the reduction can be carried out by treatment with sodium borohydride or sodium cyanoborohydride under standard conditions.

In process (h), the reaction may be carried out under conditions analogous to those described in process (e) above.

Salts of the compounds of formula (I) may be prepared by reacting the free base or a salt, enantiomer, tautomer or protected derivative thereof with one or more equivalents of the respective acid. This reaction may be carried out in a solvent or medium in which the salt is insoluble or in a solvent in which the salt is soluble, such as e.g. water, dioxane, ethanol, tetrahydrofuran or diethyl ether, or in a mixture of solvents that can be removed in vacuo or lyophilized. This reaction may be performed as a metathetical exchange reaction or may be performed on an ion exchange resin.

The compound of formula II may be prepared by reduction of the corresponding compound of formula XIII

U.

W where U, V, A and W are as defined above.

The reduction can be carried out under conditions analogous to those described above for process (f).

Some of the compounds of formula II are either known or can be prepared by conventional methods known per se. Other compounds of formula II can be prepared from known compounds with simpler substituents according to analogous procedures described above for processes (c) to (j). For example, analogously to process (j) above, it has been found advantageous to prepare some compounds of formula XIII wherein A is CH (-NXY) and X is hydrogen by reduction of the corresponding imine formed by reaction of a compound of formula NH ₂ Y with a nitrated bicyclic ketone .

Compounds of formula IV may be prepared by analogous procedures to those described for the preparation of compounds of formula II. Compounds of formula IV may be converted to the corresponding compounds of formula II by treatment with a base. Compounds of formula II may be converted to the corresponding compounds of formula IV by treatment with protic acid HA, e.g. one of the acids listed above.

Compounds of formula III are either known or can be prepared by known methods. So eg. compounds of formula III in which L represents thioalkyl may be prepared by treatment with the corresponding thiamide of formula XIV

WITH

wherein D is as defined above, with an alkyl iodide.

Compounds of formula VII, VIII, IX, X, XI, and XII can be prepared by analogous procedures to those described for the preparation of compounds of formula I. These compounds can be readily prepared from compounds with simpler substituents by conventional methods, e.g. amide formation (VII, X) by reaction of an amine with a carboxylic acid or an activated derivative thereof, or imine formation (IX, XI, XII) by reaction of an amine with an aldehyde.

Compounds of formula V, VI, XIII and XIV are either known or can be prepared by conventional methods per se.

The skilled artisan will appreciate that it may be desirable to protect an amine or other reactive group with the protecting group described in Protecting Groups in Organic Synthesis 2nd Edition (1991) by Greene and Wuts. The amino protecting groups which may be mentioned include C 2 to C 7 alkoxycarbonyl, e.g. t-butyloxycarbonyl, C 8-13 phenylalkoxycarbonyl, e.g. benzyloxycarbonyl or especially trifluoroacetate. The deprotection is generally carried out by treatment with an aqueous base, acid or hydrogen.

The compounds of this invention and intermediates can be isolated from the respective reaction mixtures by conventional procedures.

The term C 1-6 alkyl refers to straight, branched, saturated, unsaturated, aliphatic or cyclic alkyl groups having 1 to 6 carbon atoms.

Compounds of formula I may exist as tautomers, enantiomers or diastereomers, all of which are included within the scope of the invention. The various optical isomers may be isolated by separation of a racemic mixture of these compounds using conventional procedures, e.g. by fractional crystallization or by HPLC. Alternatively, the individual enantiomers may be prepared by reacting an appropriate optically active starting material with conditions such that racemization is avoided.

Intermediates may also exist as enantiomers and may be used as purified enantiomers, diastereoisomers, racemates or mixtures.

The compounds of formula I exhibit useful pharmacological action on animals. In particular, these compounds have a beneficial NO-synthase inhibitory action and are expected to be useful in the treatment or prophylaxis of human diseases under conditions in which the synthesis or over-synthesis of nitric oxide is an essential component; such as e.g. hypoxia, e.g. in cardiac arrest or stroke, neurovegetative disorders including nerve degeneration and / or nerve necrosis in disorders such as hypoxia, hypoglycaemia, epilepsy, and external injuries (such as spinal cord and head injuries), hyperbaric oxygen attacks and toxicity, dementia, e.g. presenile dementia, Alzheimer's and AIDS-related dementia, Sydenham's disease (S.chorea), Parkinson's disease, Tourette's syndrome, Huntington's disease, amyotropic lateral sclerosis, Korsakoff's disease, cerebral vascular disorders, sleep disorders, depression, schizophrenia , seasonal disorders, time zone change disorders, depression or other symptoms associated with premenstrual syndrome (PMS), anxiety states and septic shock. It is expected that the compounds of Formula I will also exhibit activity in preventing and altering tolerance to opiates and diazepines, in the treatment of drug addictions, to relieve pain, and to be useful in the treatment of migraine and other vascular headaches. The compounds of the present invention may also exhibit advantageous immunosuppressive activity, may be used in the treatment or prophylaxis of inflammation, neurogenic inflammation, transient aircraft disorders including asthma and adult respiratory distress syndrome (ARDS), treatment of gastrointestinal motility disorders, cancer, induction of labor, to reduce gastric acid secretion and to increase the contractile strength of skeletal muscles.

Compounds of the general formula in the treatment of neurodegenerative prevention and altered tolerance in the treatment of drug addictions and non-reactive diseases.

I appear to be the most interesting diseases, migraines or in opiates and diazepines or in particular in the treatment of neurodege. According to another compound of the general acceptable salt for (pharmaceutical).

In an aspect of the present invention, we propose Formula I or a pharmaceutical use thereof as a pharmaceutical preparation

According to a further feature of the present invention, we propose the use of a compound of formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prophylaxis of the above diseases and conditions.

Also contemplated is a method of treating or prophylaxis of one of the aforementioned diseases or conditions, comprising administering therapeutically effective doses of a compound of Formula I, or a pharmaceutically acceptable salt thereof, to a person suffering from or susceptible to such diseases or conditions.

Of course, in the above therapeutic indications, the dosage of the compositions to be administered will vary with the compound employed, the route of administration and the treatment desired. In general, however, satisfactory results are obtained when administered in human medicine at a daily dose of between 1 mg and 2000 mg (based on solid substance).

The compounds of the formula I and their pharmaceutically acceptable salts can be used as such or in the form of the corresponding medicinal preparations for oral or parenteral administration.

According to the present invention, there is provided a pharmaceutical formulation comprising preferably less than 80% and more preferably less than 50% of a compound of formula I, or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable diluent or carrier.

Examples of suitable diluents and carriers are well known to those skilled in the art.

The nitric oxide synthetase enzyme has a number of isomeric forms, and the effect of the compounds of formula I or their pharmaceutically acceptable salts on NO synthase activity can be evaluated according to the procedure proposed by Bredt and Snyder in Proc. Nat. Acad. Sci (1990) 87, 682-685 and Forstermann et al., Eur J. Pharm. (1992) 225, 161-165. NO-synthase converts ³ HL-arginine to ³ HL-citrulline, which can be separated by cation exchange chromatography and quantified by scintillation counting.

Screen A (A) Screening of neuronal NO-synthase activity

The enzyme was isolated from the rat brain hippocampus or small brain. The small brain or hippocampus of male Sprague-Dawley rats (250-275 g) is removed after CO ₂ anesthesia and head removal. Small brain or hippocampus supernatant was prepared by homogenization in 50 mM Tris-HCl buffer with 1 mM EDTA (pH 7.2 at 25 ° C) and centrifugation for 15 minutes at 20000 g. Residual L-arginine was removed from the supernatant by chromatography on a Dowex AG-50W-X8 ion exchange column on a sodium cycle and then on a hydrogen cycle column, and then centrifuged at 1000 g for 30 seconds.

For this evaluation, 25 μΐ of the final supernatant was added to 12 tubes containing 25 μΐ of L-arginine solution (at 18 μΐ - HL HL-arginine, 96 nM ³ HL-arginine) and either 25 μΐ of assay buffer (50 mM HEPES, 1). mM EDTA,

1.5 mM CaCl ₂ , pH 7.4) or 25 μΐ of test compound in this buffer at 22 ° C. To each tube was added 75 μΐ complete assay buffer (assay b.) (50 mM HEPES, 1 mM EDTA,

1.5 mM CaCl ₂ , 1 mM DTT, 100 μ 100 NADPH, 10 μg / ml calmodulin, pH 7.4) to initiate the reaction and this reaction was stopped after 10 minutes by the addition of 2 ml termination buffer (20 mM HEPES, 2 mM EDTA pH 5.5).

The labeled L-citrulline was separated from the labeled L-arginine by chromatography on a Dowex AG-50W-X8 ion exchange column with a particle size of 200-400 mesh. 1 ml of each completed reaction was added to a separate 1 ml column and the eluate combined with the eluate. two washes with 1 ml of distilled water and 16 ml of scintillation cocktail. L-citrulline was then quantified by scintillation counting.

In a typical experiment using a small brain supernatant, basal activity was increased by 20,000 dpm / ml of the sample above the reagent blank, which was 7000 dpm / ml. N-nitro-L-arginine, which showed 60% inhibition of NO-synthase at a concentration of 1 μΜ, was used to verify the procedure.

Screen B (B) Screening of macrophage NO-synthase activity

The enzyme is prepared from cultured murine macrophage murine J774A-1 cells (obtained from the Imperial Cancer Research Fund). J774A-1 cells were cultured in Dulbecco Modified Eagles Medium (DNEM) medium supplemented with 10% fetal bovine serum, 4 mM L-glutamine and antibiotics (100 U / ml penicillin G, 100 gg / ml steptomycin and 0.25 µM) / τη1 amphotericin B). Cells were routinely cultured in 225 cm ³ flasks containing 35 ml of nutrient medium at 37 ° C and in a humidified atmosphere containing 5% CO ₂ .

NO-synthase is produced by cells in response to interferon-τ (iNFi) and lipopolysaccharide (LPS). Nutrient medium from confluent culture flasks was removed and replaced with 25 ml (into each flask) of fresh medium containing 1 µg / ml LPS and 10 units / ml INFt. After 17-20 hours of culture, the cells were harvested by scraping their layer off the surface of the flask into the culture medium. Cells were harvested after centrifugation (1000 g for 10 minutes) and the lysate was prepared by adding the centrifuged cells to a solution containing 50 mM Tris-HCl buffer (pH 7.5 at 20 ° C), 10% (v / v) glycerol, 0.1 % (v / v) Triton-X-100, 0,1 μΜ dithiothreitol and a protease inhibitor cocktail containing leupeptin (2 μ9 ^ 1), soybean trypsin inhibitor (10 μς / ιπΐ), aprotinin (5 μg / ml) and phenylmethylsulfonylfluoride (50 mg / ml).

In this evaluation, 25 μΐ of substrate cocktail (50 mM Tris-HCl (pH 7.5 at 20 ° C), 400 μΜ NADPH, 20 μΜ flavinadenine dinucleotide, 20 μΜ flavin mononucleotide, 4 μΜ tetrahydrobiopterin, 12 μΜ L-arginine and 0.025 μθί L- [ ³ H] arginine) was added to wells on a well filter plate (pore size 0.45 μΜ) containing a 25 μΐ solution of test compound in 50 mM Tris-HCl. The reaction was started by the addition of 50 μΐ of cell lysate (prepared as described above) and after incubation for one hour at room temperature, the reaction was terminated by the addition of 50 μΐ of an aqueous solution of 3 mM nitroarginine and 21 mM EDTA.

Labeled L-citrulline was separated from labeled L-arginine using a Dowex AG-50W ion exchanger. 150 μΐ of a 25% aqueous Dowex 50W (Na ⁺ Form) suspension is added to the assay and then all material is filtered onto 96 well plates. Collect 70 μΐ of the filtrate and add to the wells of the 96 well plates containing the solid scintillation agent. After drying the samples, the L-citrulline content is determined by quantitative scintillation counting.

In a typical experiment, a basal activity of 300 dpm per 70 μ 70 sample was found, and this value increased to 1900 dpm in reagent controls. Aminoguanidine, whose IC ₅₀ (50% inhibitory concentration) was 10 μΜ, was used as a standard to verify the procedure.

Screen C (C) Screening of endothelial NO-synthase activity

This enzyme can be isolated from human endothelial cells in endothelial cells. Pollock et al. (1991) These HUVEC cells with a phosphate 800 rpm.

umbilical cord veins (human umbilical (HUVEC)) according to the procedure of Proc.Nat.Acad.Sci., 88, 10480-10484.

obtained from Clonetics Corp (San Diego, CA, USA) and cultured to confluency. These cells were maintained for 35-40 passages without significantly reducing NO-synthase yield. When the cells formed, they were resuspended in Dulbecco's salt buffer, centrifuged for 10 minutes and the cell pellet was homogenized in ice-cold pH 4.2 buffer containing 50 mM Tris-HCl, 1 mM EDTA, 10 % glycerol, 1 mM phenylmethylsulfonyl fluoride, 2 μ 2 leupeptin. After centrifugation at 34000 rpm for 60 minutes, the obtained pellet was solubilized in homogenization buffer, which also contained 20 mM CHAPS. After a 30 minute incubation on ice, the suspension was centrifuged at 34000 rpm for 30 minutes. The supernatant obtained was stored at -80 C until use.

In this evaluation, 25 μΐ of the final supernatant was added to 12 tubes containing 25 μΐ of L-arginine solution (12 μΜ-'-HL-arginine, 64 nM ³ HL-arginine) and either 25 μΐ of assay buffer (50 mM HEPES, 1 mM EDTA,

1.5 mM CaCl ₂ , pH 7.4) or 25 µΐ test compound in this buffer at 22 ° C. To each tube was added 25 μΐ of complete assay buffer (assay b.) (50 mM HEPES, 1 mM EDTA,

1.5 mM CaCl ₂ , 1 mM DTT, 100 μΜ NADPH, 10 μg / ml calmodulin, 12 μΜ tetrahydrobiopterin, pH 7.4) to initiate the reaction the reaction was stopped after 10 minutes by addition of stop buffer (20 mM HEPES, 2 mM EDTA pH 5.5).

and this 2 ml

Labeled L-citrulline was separated from labeled L-arginine by chromatography on a Dowex AG-50W-X8 ion exchange column with a particle size of 200-400 mesh. Then 1 ml of each completed reaction was added to a separate 1 ml column and the eluate was combined with the eluate from two washes of 1 ml of distilled water and 16 ml of scintillation cocktail. L-citrulline was then quantified by scintillation counting.

In a typical experiment, basal activity was increased by 5,000 dpm / ml of the sample above the reagent blank that showed an activity of 1500 dpm / ml. N-nitro-L-arginine, which showed 70-90% inhibition of NO-synthase at a concentration of 1 μΜ, was used to verify the procedure.

These compounds can also be tested in ex-vivo experiments to assess the rate of their penetration into the brain.

Screen D

Ex-vivo experiments to determine neuronal NO-synthase activity

Male Sprague-Dawley rats (250-275 g) were treated intravenously with 10 mg / kg of the test compound dissolved in 0.9% saline or in saline control alone. After a predetermined time (typically 2 to 24 hours) after administration, the animals were sacrificed, the cerebellum was removed, and the supernatant was prepared and evaluated for nitrate synthetase activity according to the procedure described in Screen A.

In a further validation experiment, a portion of the cerebellum supernatant was loaded onto a column packed with 2'-5'-ADP Sepharose (which binds NO-synthase) and then eluted with NADPH. The eluate obtained was tested for NO-synthase activity according to the procedure described in Screen A.

Compounds that penetrate the rat brain and inhibit neuronal NO-synthase exhibit a decrease in NO-synthase activity in both the prepared supernatant and in the eluate from the column packed with 2'-5'-ADP Sepharose.

When screening for NO-synthase activity, the activity of the individual compounds was expressed as IC ₅₀ (the concentration of the substance that caused a 50% decrease in activity when evaluated). IC ₅₀ values of test substances were initially determined by the inhibitory activity of 1, 10 and 100 μΜ solutions of these compounds. Compounds that inhibited this enzyme by at least 50% at a concentration of 10 μΜ were retested at the corresponding concentrations which allowed a more accurate determination of the IC 50 value.

In the above-mentioned screening A (evaluation of activity against the neuronal isoform of NO-synthase), the compound of Example 1 below gave an IC ₅₀ value of less than 10 µΜ, indicating possible therapeutic utility. The screening of B and C (to assess the activity of macrophage and endothelial isoforms of nitric oxide synthetase) the compound gave the following Examples 1 IC _5Q values greater than ten times the value of the previously Screen A indicating the preferred selectivity.

The compounds of Examples 2-9, 10 (a) - (f), 11-13 and 19-24 were also tested in screening A and also gave an IC ₅₀ value of less than 10 µΜ. Thus, useful therapeutic activity is also expected with these agents.

The compounds of formula I and their pharmaceutically acceptable salts have the advantage of being less toxic, more potent, more selective, having a longer duration of action, having a broader spectrum of activity, more potent, causing less side effects, more readily absorbed or other suitable pharmaceuticals. properties as compared to substances previously known and used in the therapeutic areas mentioned above.

The compounds of formula (I) and their pharmaceutically acceptable salts may also have the advantage of having a more selective action on the neuronal isoform of NO-synthase, and therefore can be expected to have advantageous therapeutic utility with reduced side effects for inhibiting other isoforms.

DETAILED DESCRIPTION OF THE INVENTION

The invention is illustrated by the following examples:

Example 1

N - ((2- (Phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dioxalate (a) 5-Nitro-2-indanone

The compound was prepared by the procedure of Heusler and Schieffer Ber., (1899) 32.33.

(b) 5-Nitro-2- (phenylmethyl) aminoindan

The flask was charged with: 5-nitro-2-indanone (1.48 g,

8.36 mmol), benzylamine (4.40 mL, 41.8 mmol), acetic acid (15.0 mL), 4 A molecular sieve (20 mL), THF (15 mL) and MeOH (15 mL) and this the mixture was cooled to 0 ° C. Sodium borohydride (1.05 g, 16.7 mmol) was then added portionwise over 5 minutes. The mixture was stirred for 14 hours, filtered through celite and concentrated to a syrup. The mixture was basified with 2N NaOH and extracted with ether (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, concentrated and chromatographed on silica gel (3% methanol / methylene chloride). Yield of 5-nitro-2- (phenylmethyl) -aminoindan: (1.18 g, 53%); Mass Spec: (M + H) ⁺ = 269.

(c) 2- (5-Nitroindanyl) -N- (phenylmethyl) trifluoroacetamide

To a stirred solution of 5-nitro-2- (phenylmethyl) aminoindan (1.18 g, 4.40 mmol) and triethylamine (0.61 mL, 4.40 mmol) in methylene chloride (50 mL) was added dropwise trifluoroacetic anhydride (0). , 63 ml, 4.40 mmol). After stirring for 1 minute, the reaction mixture was poured into water and extracted with methylene chloride (3 x 20 mL). The combined extracts were washed with water, dried over magnesium sulfate and filtered through a short pad of silica gel (20% ethyl acetate / hexane) to give 2- (5-nitroindanyl) -N- (phenylmethyl) trifluoroacetamide: (1.17 g, 73%) ); Mass spec.

(M + H) < ^{+ &gt}; = 365.

- 22 (d) 2- (5-Aminoindanyl) -N- (phenylmethyl) trifluoroacetamide

To a stirred solution of 2- (5-nitroindanyl) -N- (phenylmethyl) trifluoroacetamide (1.17 g, 3.21 mmol) in THF / MeOH (100 mL, 1: 1) was added a catalytic amount of 10% Pd / C. The mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to give 2- (5-aminoindanyl) -N- (phenylmethyl) trifluoroacetamide which was TLC homogeneous and used immediately in reaction step (f).

(e) S-methyl-2-thiophenethiocarboximide hydroiodide

To a solution of 2-thiophenecarboxthioamide (Maybridge Chemical) (11.1 g) in acetone (60 mL) was added iodomethane (13.4 g). After 6 hours at 22 ° C, a yellow solid formed by filtration, washed with acetone (2 x 25 mL) and dried to give 18.45 g of S-methyl-2-thiophenethiocarboximide hydroiodide, m.p. 195 “C. (F) N - ((2- (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dioxalate

To a solution of 2- (5-aminoindanyl) -N- (phenylmethyl) trifluoroacetamide (1.0 g, 3.0 mmol) in isopropanol (6 mL) / DMF (0.5 mL) was added S-methyl-2. -thiophenethiocarboximide hydroiodide (0.85 g, 3.0 mmol). The mixture was stirred for 14 h, diluted with methanol (6 mL) and 2N NaOH (6 mL) and heated to 50 ° C for 0.5 h. This mixture was poured into aqueous and extracted with ethyl acetate (3 x 30 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and chromatographed on silica gel (20% methanol / methylene chloride) to afford the title compound as the free base. Treatment with IPA / oxalic acid gave N - ((2- (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dioxalate as a white solid: (0.47 g, 30%); mp 130-135 ’C.

Example 2

N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide (a) 7-Nitro-3,4-dihydro-2 (1H) -naphthalenone

This compound was prepared according to the method described in J. Med.Chem. (1989), 32,2128.

(b) 7-Nitro-2 - ((phenylmethyl) amino) -1,2,3,4-tetrahydronaphthalene

Charge the flask with 7-nitro-3,4-dihydro-2 (1H) -naphthalenone (1.50 g, 7.85 mmol), benzylamine (4.30 mL, 39.3 mmol), acetic acid (8, 0 mL), 4 A molecular sieve (20 mL), THF (15 mL) and MeOH (15 mL) and the mixture was cooled to 0 ° C. Sodium cyanoborohydride (0.99 g, 15.7 mmol) was then added portionwise over 5 minutes. The mixture was stirred for 14 hours, filtered through celite and concentrated to a syrup. It was then basified with 2N NaOH and extracted with ether (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and chromatographed on silica gel (3% methanol / methylene chloride) to give 7-nitro-2 - ((phenylmethyl) amino) -1,2,3,4-tetrahydronaphthalene (2, 3, 4-tetrahydronaphthalene). 10 g, 95%), mass spec (M + H) ⁺ = 283.

(c) 2- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide

I

To a stirred solution of 7-nitro-2 - ((phenylmethyl) amino) -1,2,3,4-tetrahydronaphthalene (2.10 g, 7.45 mmol) and triethylamine (1.07 mL, 7.45 mmol) in methylene chloride (50 mL) was added dropwise trifluoroacetic anhydride (1.05 mL, 7.45 mmol). After stirring for 1 minute, the mixture was poured into water and extracted with methylene chloride (3 x 20 mL). The combined extracts were then washed with water, dried over magnesium sulfate and filtered through a short pad of silica gel (20% ethyl acetate / hexane) to give 2- (7-nitro- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) ) trifluoroacetamide: (2.55 g, 90%); Mass spec. (M + H) < ^{+ &gt}; = 379.

(d) 2- (7-Amino- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide

To a stirred solution of 2- (7-nitro- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide (2.55 g, 6.75 mmol) in THF / MeOH (100 mL, 1 mL). 1) a catalytic amount of 10% PD / C was added. This mixture was hydrogenated at 50 psi (1 psi = 0.689.10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to give 2- (7-amino- (1,2,3,4-tetrahydronaphthyl)). -N- (phenylmethyl) trifluoroacetamide, which was homogeneous by TLC and used immediately in the next reaction step.

(e) N - ((2-phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide

To a solution of 2- (7-amino- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide (2.11 g, 6.07 mmol) in isopropanol (10 mL) was added S- methyl 2-thiophenethiocarboximide hydroiodide (1.72 g, 6.07 mmol). The mixture was stirred for 14 h, diluted with methanol (6 mL) and 2N NaOH (6 mL) and heated to 50 ° C for 0.5 h. The mixture was poured into water and extracted with ethyl acetate (3 x 30 mL). ml). The combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to give a solid which, after recrystallization (methylene chloride / hexane), gave N - ((2-phenylmethyl) amino) -1,2,3,4-tetrahydronaphtha. -7-yl) -2-thiophenecarboximidamide as a white solid: (0.66 g, 30%); mp 119-120 ’C.

Example 3

N - ((2-Amino) -1,2,3,4-tetrahydronaphthal-7-yl) -2-thiophenecarboximidamide dioxalate (a) 7-Nitro-2-amino-1,2,3,4-tetrahydronaphthalene hydrochloride

7-Nitro-1-tetralone (1.50 g,

7.85 mmol), ammonium acetate (6.05 mL, 78.5 mmol), acetic acid (8.0 mL), 4 A molecular sieve (20 mL), THF (15 mL) and MeOH (15 mL), and this mixture was cooled to 0 ° C. Sodium cyanoborohydride (0.99 g, 15.7 mmol) was added portionwise over 5 minutes. The resulting mixture was stirred for 1 hour, filtered through celite and concentrated to a syrup. The mixture was basified with 2N NaOH and extracted with ether (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. The compound was isolated as the hydrochloride: 7-nitro-2-amino-1,2,3,4-tetrahydronaphthalene hydrochloride: (1.00 g, 56%), mp> 300 ° C.

(b) 2- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide

To a stirred solution of 7-nitro-2-amino-1,2,3,4-tetrahydronaphthalene hydrochloride (1.00 g, 4.39 mmol) and triethylamine (1.22 mL, 8.77 mmol) in methylene chloride (50 mL) Trifluoroacetate hydride (0.62 mL, 4.39 mmol) was added dropwise. After stirring for 1 minute, the mixture was poured into water and extracted with methylene chloride (3 x 20 mL). The combined extracts were then washed with water, dried over magnesium sulfate and filtered through a short pad of silica gel (20% ethyl acetate / hexane) to give 2- (7-nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide: (0.78 g, 62%); Mass spec. (M + H) < ^{+ &gt}; = 289.

(c) 2- (7-Amino- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide

To a stirred solution of 2- (7-nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide (0.76 g, 2.21 mmol) in THF / MeOH (100 mL, 1: i) was added added a catalytic amount of 10% Pd / C. This mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to give 2- (7-amino- (1,2,3,4-tetrahydronaphthyl)). The N-trifluoroacetamide which was homogeneous by TLC and used immediately in the next reaction step.

(d) N - ((2-Amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dihydrobromide

To a solution of 2- (7-amino- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide (0.70 g, 2.71 mmol) in isopropanol (10 mL) was added S-methyl-2- thiophenothiocarboximide hydroiodide (0.77 g,

2.71 mmol). The mixture was stirred for 14 h, diluted with methanol (6 mL) and 2N NaOH (6 mL) and heated to 50 ° C for 0.5 h. The mixture was poured into water and extracted with ethyl acetate (3 x 30 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, concentrated and chromatographed on silica gel (20% methanol / methylene chloride) to give an oily product which was converted to the dihydrobromide: N - ((2-amino) -1,2) 3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dihydrobromide: (0.37 g, 32%); decomposition> 210 ’C.

Example 4

N - ((1-Amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dioxalate (a) 7-Nitro-1-amino-1,2,3,4-tetrahydronaphthalene

7-Nitro-1-amino-1,2,3,4-tetrahydronaphthalene was also prepared as 7-Nitro-2-amino-1,2,3,4-tetrahydronaphthalene. This compound was isolated as the hydrochloride: (0.30 g, 12%); > 300 300 C.

(b) 1- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide

1- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide was also prepared as 2- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide (0.35 g, 95%); weight. Spectra. (M + H) < ^{+ &gt}; = 289.

(c) 1- (7-Amino- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide 1- (7-Amino- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide also prepared as 2- (7-amino- (1,2,3,4-tetrahydronaphthyl)) - N-trifluoroacetamide and was used immediately in the next reaction.

(d) N - ((1-Amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximide 27-amide dioxalate

N - ((1-amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dioxalate was also prepared as N - ((2-amino) -1,2,3,4- tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dihydrobromide except that it was isolated as the oxalic acid salt: (0.18 g, 33%); decomposition > 155 ° C.

Example 5

N - ((2-Amino) -indan-5-yl) -2-thiophenecarboximidamide dioxalate (a) 5-Nitro-2-aminoindan hydrochloride

To 2-aminoindan hydrochloride (19.11 g, 0.112 mol) at 0 ° C was added sulfuric acid (60 mL) followed by potassium nitrate (11.84 g, 0.117 mol). This mixture was allowed to warm to room temperature, stirred for an additional 2 hours and then poured onto ice / 50% NaOH (total 500 mL). The mixture was extracted with ether (3 x 200 mL), the combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to an oily product which was converted to the hydrochloride. Recrystallization from isopropanol / methanol gave 5-nitro-2-aminoindan hydrochloride: (14.58 g, 60%); mp > 300 ° C.

(b) 2- (5-Nitroindanyl) -N-trifluoroacetamide

To a stirred solution of 5-nitro-2-aminoindan hydrochloride (1.00 g, 5.89 mmol) and triethylamine (0.82 mL, 5.89 mmol) in methylene chloride (50 mL) was added dropwise trifluoroacetic anhydride (0.83 ml, 5.89 mmol). After stirring for 1 minute, the mixture was poured into water and extracted with methylene chloride (3 x 20 mL). The combined extracts were then washed with water, dried over magnesium sulfate and filtered through a short pad of silica gel (20% ethyl acetate / hexane) to give 2- (5-nitroindanyl) -N-trifluoroacetamide: (1.51 g, 93%); mp 153-154 ’C.

(c) 2- (5-Aminoindanyl) -N-trifluoroacetamide

To a stirred solution of 2- (5-nitroindanyl) -N-trifluoroacetamide (0.58 g, 2.25 mmol) in THF / MeOH (100 mL, 1: 1) was added a catalytic amount of 10% Pd / C. This mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to give 2- (5-Aminoindanyl) -N-trifluoroacetamide homogeneous by TLC and used immediately in the next reaction step.

(d) N- ((2-Amino) indan-5-yl) -2-thiophenecarboximidamide dioxalate

To a solution of 2- (5-aminoindanyl) -N-trifluoroacetamide (0.52 g,

2.25 mmol) in isopropanol (6 mL) and DMF (0.5 mL) were added S-methyl-2-thiophenethiocarboxymide hydroiodide (0.64 g, 2.25 mmol). The mixture was stirred for 14 hours, diluted with methanol (6 mL) and 2 N NaOH (6 mL) and heated to 50 ° C for 0.5 hour. The mixture was poured into water and extracted with ethyl acetate (3 x 30 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and chromatographed on silica gel (20% methanol / methylene chloride) to give the title compound as the free base. Treatment with IPA / butyric acid gave N - ((2-amino) indan-5-yl) -2-thiophenecarboximidamide dioxalate as a white solid: (0.60 g, 50%); b. t. decomposition 70 ’C.

Example 6

N- ((2-) Methyl) (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dihydrobromide (a) 5-Nitro-2- (phenylmethyl) aminoindan hydrochloride

To 5-nitro-2-aminoindan hydrochloride (3.00 g, 14.00 mmol) in DMF (60 mL) was added triethylamine (4.07 mL, 29.40 mmol) followed by benzyl bromide (1.74 mL, 14 mL). , 68 mmol). The mixture was stirred at room temperature for 1 hour, poured into water (200 mL) and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered through a short pad of silica gel, and concentrated to a syrup. The subtitle substance was obtained as the hydrochloride:

(2.29 g, 54%); mp decomposition 266 ’C.

(b) 5-Nitro-2- (methyl) (phenylmethyl) aminoindan hydrochloride

To 5-nitro-2- (phenylmethyl) aminoindan hydrochloride (2.29 g,

Potassium carbonate (2.60 g, 18.80 mmol) was added in DMF followed by methyl iodide (0.47 mL, 7.52 mmol). This mixture was warmed to room temperature, stirred for 16 hours, poured into water (400 mL) and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered through a short pad of silica gel, and concentrated to a syrup. The title compound was obtained as the hydrochloride: (1.08 g, 45%); mp decomposition 280 ’C.

(c) 5-Amino-2- (methyl) (phenylmethyl) aminoindanediol hydrochloride

To 5-nitro-2- (phenylmethyl) aminoindan hydrochloride (1.08 g,

3.39 mmol) in 85% acetic acid in water was added zinc powder (3.0 g). The mixture was stirred for 1 minute, filtered through celite and concentrated. The resulting concentrate was neutralized with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate and evaporated to a syrup. IPA / HCl was added to the obtained oily product, the reaction mixture was concentrated and used in the next reaction step.

(d) N- ((2-) Methyl) (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dihydrobromide

To 5-amino-2- (methyl) (phenylmethyl) aminoindanedihydrochloride in DMF (10 mL) was added S-methyl-2-thiophenethiocarboximide hydroiodide 0.98 g, 3.45 mmol) and pyridine (0.27 mL, 3.29 mmol). The mixture was stirred for 14 h, poured into water with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and chromatographed on silica gel (10% methanol / methylene chloride) to give the title compound as the free base. Treatment with IPA / oxalic acid gave N - ((2-) Methyl) (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide dihydrobromide as a white solid: (0.43 g, 25%); mp 196-200 ’C.

Example 7

N - ((1-amino) indan-6-yl) -2-thiophenecarboximidamide dihydrochloride (a) 6-Nitro-1-aminoindan hydrochloride

1-Aminoindane (10.0 g, 75.08 mmol) was added to concentrated sulfuric acid (40 mL) at 0 ° C. This mixture was allowed to warm to room temperature to aid dissolution and then cooled to 0 ° C. . Potassium nitrate (7.60 g, 75.08 mmol) was added portionwise and the mixture was stirred at room temperature for 1 hour, then poured onto ice with 50% NaOH. The resulting aqueous solution was extracted with chloroform (3 x 100 mL). The combined extracts were washed with water, decolourised with charcoal, dried with magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / HCl gave the title compound: (6.90 g, 43%); b. t. decomposition 280 ’C.

(b) 6-Amino-1-aminoindan hydrochloride

To a solution of 6-nitro-1-aminoindan hydrochloride (1.00 g, 4.66 mmol) in MeOH (100 mL) was added a catalytic amount of 10% Pd / C. This mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated. 6-Amino-1-aminoindan hydrochloride was obtained which was homogeneous by TLC and used immediately in the next reaction step.

(c) N - ((1-amino) indan-6-yl) -2-thiophenecarboximidamide dihydrochloride

To 6-amino-1-aminoindan hydrochloride (0.74 g, 4.01 mmol) in DMFA / IPA (4 mL, 1: 1) was added S-methyl-2-thiophenethio carboximide hydroiodide (1.26 g, 4, 41 mmol). This mixture was heated to 50 ° C, stirred for 16 hours, poured into water with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / oxalic acid gave N - ((1-amino) indan-6-yl) -2-thiophenecarboximidamide dihydrochloride as a white solid: (0.79 g, 60%) m.p .: decomposition > 200 ° C.

Example 8

N - ((1- (phenylmethyl) amino) indan-6-yl) -2-thiophenecarboximidamide dioxalate (a) 6-Nitro-1- (phenylmethyl) aminoindan hydrochloride

To 6-nitro-1-aminoindan hydrochloride (1.90 g, 8.85 mmol) in DMF (30 mL) was added triethylamine (2.50 mL, 18.06 mmol) followed by benzyl bromide (1.07 mL, 9, 03 mmol). This mixture was warmed to room temperature, stirred for 3 hours, poured into water (100 mL) and extracted with ethyl acetate (3 x 70 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered through a short pad of silica gel, and concentrated to an oil. The title compound was isolated as the hydrochloride salt: (1.34 g, 50%); mp 234-235 “C.

(b) 6-Amino-1- (phenylmethyl) aminoindan hydrochloride

To 6-nitro-1- (phenylmethyl) aminoindan hydrochloride (1.34 g,

4.40 mmol) in MeOH (100 mL) was added a catalytic amount of 10% Pd / C. This mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated. 6-amino-1- (phenylmethyl) aminoindan hydrochloride was obtained which was homogeneous by TLC and used immediately in the next reaction step.

(c) N - ((1- (phenylmethyl) amino) indan-6-yl) -2-thiophenecarboximidamide dioxalate

To 6-amino-1- (phenylmethyl) aminoindan hydrochloride (1.21 g,

4.40 mmol) in DMF (20 mL) was added S-methyl-2-thiophenethiocarboximide hydroiodide (1.38 g, 4.84 mmol). This mixture was heated to 50 ° C for 16 hours, poured into water with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to an oily product. Treatment with IPA / oxalic acid gave N - ((1- (phenylmethyl) amino) indan-6-yl) -2-thiophenecarboximidamide dioxalate as a white solid: (1.06 g, 46%); mp .: decomposition> 120 C.

Example 9

N - ((2 - ((3-Chlorophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine (a) 2 - ((3-Chlorophenyl) carbonyl) amino-6-nitroindane

To 2-amino-6-nitroindan hydrochloride (1.5 g, 7.0 mmol) in methylene chloride (50 mL) at 0 ° C was added triethylamine (2.1 mL, 15.0 mmol) followed by 3-chlorobenzoyl chloride ( 1.0 ml, 7.5 mmol). This mixture was immediately poured into water and the layers were separated. The aqueous layer was extracted with methylene chloride (2 x 20 mL) and the combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to an oily product which was homogeneous by TLC and used immediately in the next step: Mass. (M + H) < ^{+ &gt}; = 317.

(b) 2 - ((3-Chlorophenyl) methyl) araino-6-nitroindane

To 2 - ((3-chlorophenyl) carbonyl) amino-6-nitroindane (2.2 g, 7.0 mmol) in THF (75 mL) was added dropwise BH ₃ .THF (1.0 M, 35 mL, 35 mmol). The mixture was heated at reflux for 12 hours, cooled to 0 ° C, 4N HCl (60 mL) was added and the mixture was refluxed for 1 hour. The resulting solution was evaporated to an oily product, basified with 50% NaOH and extracted with methylene chloride (3 x 20 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oil. Treatment with IPA / HCl gave 2 - ((3-chlorophenyl) methyl) amino-6-nitroindane: (2.1 g, 88% over two steps); mp 234-237 ° C.

(c) 2 - ((3-Chlorophenyl) methyl) amino-6-aminoindan

To 2 - ((3-chlorophenyl) methyl) amino-6-nitroindan hydrochloride (2.1 g, 6.13 mmol) in 85% AcOH / H ₂ O (40 mL) was added zinc metal (1.6 g, 24 mL). , 5 mmol). The mixture was stirred for 5 minutes, filtered through celite and concentrated to an oil. The oil was poured into alkaline water and extracted with chloroform (3 x 20 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to an oil. Treatment with IPA / HCl gave 2 - ((3-chlorophenyl) methyl) amino-6-aminoindan: (1.5 g,

%); mp higher than 270 ’C.

(d) N - ((2 - ((3-Chlorophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine

A mixture of 2 - ((3-chlorophenyl) methyl) amino-6-aminoindanedihydrochloride (1.5 g, 4.2 mmol), S-methyl-2-thiophenethiocarboximide hydroiodide (1.3 g, 4.6 mmol) and pyridine (0). (34 mL, 4.2 mmol) in DMF (10 mL) was stirred for 24 hours. The mixture was then poured into water, basified with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and chromatographed on silica gel (12% methanol / methylene chloride) to give a colorless oil. Treatment with IPA / HCl gave N - ((2 - ((3-chlorophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxymidine: 0.75 g, 40%); mp

297-299.

Example 10

The following compounds were prepared according to the procedure of Example 9:

(a) N- ((2 - ((2-Methylphenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine: bt183 ° C (b) N - ((2 - ((3-Methylphenyl) methyl) (amino) indan-5-yl) -2-thiophenecarboxamidine: bt 195 ° C (g) N - ((2 - ((4-Methylphenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine: bt182 C (d) N - ((2 - ((Ethyl) amino) indan-5-yl) -2-thiophenecarboxamidine: bt236-238 C (e) N - C (2 - (((4-phenyl)) phenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine:

m.p.182 ´ C (f) N - ((2 - (((4-Hexyl) phenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine:

mp 125 ° C (g) N - ((2 - ((3-Bromophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine:

m.p.182 ° C

Example 11

N - ((2 - ((3-Chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphth-7-yl-2-thiophenecarboximidamide (a) 7-Nitro-2 - (((3-chlorophenyl)) methyl) amino) -1,2,3,4-tetrahydronaphthalene

A mixture of 7-nitro-3,4-dihydro-2 (1H) -naphthalenone (1.50 g,

7.85 mmol), 3-chlorobenzylamine (4.70 mL, 39.3 mmol), acetic acid (6.0 mL), 4 A molecular sieve (20 mL), THF (15 mL) and MeOH (15 mL) was placed in a flask and cooled to 0 ° C. To this mixture, sodium cyanoborohydride (0.99 g, 15.7 mmol) was added portionwise over 5 minutes. The mixture was stirred for 14 hours, filtered through celite and concentrated to a syrup. It was then basified with 2N NaOH and extracted with ether (3x

35 x 50 ml). The combined extracts were washed with water, dried over magnesium sulfate, filtered, concentrated and chromatographed on silica gel (3% methanol / methylene chloride) to give a colorless oil. Treatment of this oil with IPA / HCl gave 7-nitro-2 - (((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene hydrochloride: (1.34 g, 50%); Mass spec. (M + H) < ^{+ &gt}; = 317.

(b) 7-Amino-2 - (((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene

To 7-nitro-2 - (((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene hydrochloride (1.34 g, 3.80 mmol) in 85% AcOH / H ₂ O (75 mL) zinc metal (2.48 g, 38.0 mmol) was added. The mixture was stirred for 5 minutes, filtered through celite and evaporated to an oil. The resulting oil was extracted with chloroform (washed 3 times with water, dried over sulfate, poured into alkaline water and 20 mL). The combined extracts were MgSO4, filtered and concentrated to an oily product. Treatment with IPA / HCl gave 7-amino-2 - (((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene (1.4 g, 99%); weight. Spectra. (M + H) < ^{+ &gt}; = 288.

(c) N - ((2 - ((3-Chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphth-7-yl-2-thiophenecarboximidamide

A mixture of 7-amino-2 - (((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene dihydrochloride (1.32 g, 3.70 mmol), S-methyl-2-thiophenethiocarboximide hydroiodide (1) , 3 g, 4.6 mmol) and pyridine (0.30 mL, 3.7 mmol) in DMF (15 mL) were stirred for 24 hours. This mixture was then poured into water, basified with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / oxalic acid gave N - ((2 - ((3-chlorophenyl) methyl) amino) -1,2,3,4-tetrahydronaphth-7-yl-2-thiophene-carboximidamide dioxalate: (0.71) g, 33%), decomposition above 100 C.

- 36 Example 12

N - ((2- (phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphth-7-yl-2-thiophenecarboximidamide (a) 7-Nitro-2 - ((phenylmethyl) methyl) amino) -1 , 2,3,4-tetrahydronaphthalene

To a stirred solution of 7-nitro-2 - ((phenylmethyl) amino) -1,2,3,4-tetrahydronaphthalene (1.5 g, 5.4 mmol) in DMF (30 mL) was added potassium carbonate (1.5 mL). g, 10.8 mmol) and methyl iodide (0.36 mL, 5.8 mmol). The mixture was stirred for 24 hours, poured into water and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / HCl gave 7-nitro-2 - ((phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene hydrochloride: (0.89 g, 50%); Mass spec. (M + H) < ^{+ &gt}; = 297.

(b) 7-Amino-2 - ((phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene

To 7-nitro-2 - ((phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphthalene hydrochloride (0.89 g, 2.7 mmol) in 85% AcOH / H ₂ O (75 mL) zinc metal (3.5 g, 54.0 mmol) was added. The mixture was stirred for 5 minutes, filtered through celite and evaporated to an oily product. This oily product was poured into alkaline water and extracted with chloroform (3 x 20 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / HCl gave 7-amino-2 - ((phenylmethyl) methylanino) -1,2,3,4-tetrahydronaphthalene: (0.81 g, 88%); Mass spec. (M + H) < ^{+ &gt}; = 267.

(c) N- ((2- (Phenylmethyl) methyl) amino) -1,2,3,4-trihydronaphth-7-yl-2-thiophenecarboxymidamide

A mixture of 7-amino-2 - ((phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphthalenedihydrochloride (0.81 g, 2.4 mmol), S-methyl-2-thiophenothiocarboximide hydroiodide (0.74 g) , 2.6 mmol) and pyridine (0.19 mL, 2.4 mmol) in DMF (15 mL) were stirred for 24 hours. The mixture was then poured into water, basified with 2N NaOH and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, concentrated and chromatographed on silica gel (15% MeOH / methylene chloride). Concentration of the fraction gave a solid which, after recrystallization from ethyl acetate / hexane, gave N - ((2-phenylmethyl) methyl) amino) -1,2,3,4-tetrahydronaphth-7-yl-2-thiophenecarboximidamide dihydrochloride: ( 0.14 g, 16%); mp M.p. 176-178 ° C.

Example 13

N - ((1- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide (a) 7-Nitro-1 - ((phenylmethyl) amino) -1,2, 3,4-tetrahydronaphthalene

7-Nitro-1-tetralone (2.0 g, 10.5 mmol), benzylamine (1.2 mL, 10.5 mmol) and titanium isopropoxide (3.9 mL, 13.1 mmol) were mixed and stirred for 1 day. hours. This mixture was then diluted with absolute ethanol (12 mL), sodium cyanoborohydride (0.44 g, 7.0 mmol) was added and stirred for 20 hours. The solid (soilds?) Was filtered off and washed with ethanol. The ethanol was concentrated and the remaining oily product was used immediately in the next reaction: mass spec (M + H) ⁺ = 283.

(b) 1- (7-Nitro- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide ¹

To a stirred solution of 7-nitro-1 - ((phenylmethyl) amino) -1,2,3,4-tetrahydronaphthalene (2.96 g, 10.50 mmol) and diethylamine (1.46 mL, 10.50 mmol) in methylene chloride (50 mL) was added dropwise trifluoroacetic anhydride (1.46 mL, 10.50 mmol). After stirring for 1 minute, the mixture was poured into water and extracted with methylene chloride (3 x 20 mL). The combined extracts were washed with water, dried over magnesium sulfate and filtered through a short pad of silica gel (20% ethyl acetate / hexane) to give 1- (7-nitro (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide :

(1.90 g, 48%, two steps); Mass spec. (M + H) < ^{+ &gt}; = 379.

(c) 1- (7-Amino- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide

To a stirred solution of 1- (7-nitro- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide (1.91 g, 5.05 mmol) in THF / MeOH (100 mL, 1: 1) a catalytic amount of 10% Pd / C was added. This mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through. celite and concentrated to give 1- (7-amino- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide which was homogeneous by TLC and used immediately in the next reaction step .

(d) N - ((1-Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide

To a solution of 1- (7-amino- (1,2,3,4-tetrahydronaphthyl)) - N- (phenylmethyl) trifluoroacetamide (1.76 g, 5.05 mmol) in isopropanol (10 mL) was added S methyl 2-thiophenethiocarboximide hydroiodide (1.44 g, 5.05 mmol). The resulting mixture was stirred for 14 h then diluted with methanol (6 mL) and 2N NaOH (6 mL) and heated to 50 ° C for 0.5 h. Then the mixture was poured into water and extracted with ethyl acetate (3 x 30 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. Treatment with IPA / HBr gave N - ((2- (phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide dihydrobromide as a white solid: (0.53 g, 20 %); mp 260-262 ° C.

Example 14

N - ((1- (Phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide (a) 6-Acetamido- (1 - ((phenyl) methyl) amino) indane

6-Acetamido-1-indanone (5.0 g, 27.6 mmol), benzylamine (3.1 mL, 27.9 mmol) and titanium isopropoxide (10.2 mL, 34.5 mmol) were mixed and stirred for 1 day. hours. The resulting mixture was diluted with water. ··, · • ï -:::.

39 with absolute ethanol (30 mL) was added sodium cyanoborohydride (1.2 g, 19.3 mmol) and stirred for 20 hours. The solid was filtered and washed with ethanol. The ethanol was concentrated and the remaining oil was dissolved in ethyl acetate and extracted with 1N HCl (3 x 50 mL). The aqueous layer was neutralized with 2N NaOH and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and evaporated to give an oily product which was used without further purification in the next step.

(b) 6-Amino- (1 - ((phenyl) methyl) amino) indane

6-Acetamido- (1 - ((phenyl) methyl) amino) indane was refluxed with 4N HCl (50 mL) for 20 minutes, then cooled and extracted with ethyl acetate (3 x 50 mL). The aqueous layer was neutralized with 2N NaOH and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over MgSO4

magnesium, filtered, and concentrated to an oily product. This oil was dissolved in IPA and treated with IPA / HCl to afford the dihydrochloride: (2.0 g, 24% over two steps); m.p .; decomposition above 250 ° C

(c) N - ((1- (Phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide

To 6-amino- (1 - ((phenyl) methyl) amino) indane dihydrochloride (2.0 g, 6.4 mmol) in DMF (20 mL) was added S-methyl-2-thiophenothiocarboximide hydroiodide (2.2 g). , 7.7 mmol) and pyridine (0.57 mL, 7.1 mmol). The resulting mixture was stirred at 50 ° C for 20 hours, then poured into alkaline water and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, concentrated and chromatographed on silica gel (6% methanol / methylene chloride). The extracts were concentrated to an oily product, which was then dissolved in methanol, IPA / HCl and ether were added. The solids were collected by filtration and washed with ether: (1.1 g, 40%); m.p .: decomposition above 180 ’C.

Example 15

The following compound was prepared according to the method of Example 14:

N - ((1- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-6-yl) -2-thiophenecarboximidamide: m.p.: decomposition above 200 ° C.

Example 16

N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-furanecarboximidamide (a) 2 - ((Phenyl) carbonyl) amino-7-nitrotetralin

To 2-amino-7-nitrotetralin (2.8 g, 14.5 mmol) in THF (50 mL) and 10% potassium carbonate solution (100 mL) was added benzoyl chloride (1.7 mL, 15.3 mmol). After the addition was complete, the reaction mixture was diluted with water to a volume of 250 mL. The precipitated solids were collected by filtration, washed with water and dried in vacuo: (4.2 g, 98%), m.p. 194-198 ’C.

(b) 2 - ((Phenyl) methyl) amino-7-nitrotetraline hydrochloride

To 2 - ((phenyl) carbonyl) amino-7-nitrotetralin (4.2 g, 14.1 mmol) in anhydrous THF (100 mL) was added borane-THF (49.3 mL, 1M THF, 49.3 mmol). ). This mixture was heated to reflux for 5 hours, cooled to 0 ° C and quenched by dropwise addition of 4N HCl. The mixture was then refluxed for 1 hour, concentrated in vacuo and the solid was filtered (washed with water) and dried in vacuo: (3.5 g, 78%), m.p. above 300 ’C.

(c) 2 - ((Phenyl) methyl) amino-7-aminotetraline hydrochloride

To a stirred solution of 2 - ((phenyl) methyl) amino-7-nitrotetralin (2.0 g, 6.3 mmol) in MeOH (100 mL) was added a catalytic amount of 10% Pd / C. This reaction mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to an oily product which was homogeneous by TLC and used immediately in the next reaction step.

- 4ί (d) N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-furancarboximidamide

To 2 - ((phenyl) methyl) amino-7-aminotetraline hydrochloride (1.8'g, 6.3 mmol) in DMF (20 mL) was added S-methyl-2-furanthiocarboximide hydroiodide (2.0 g, 7.5 mmol). The mixture was stirred for 2 hours at 45 ° C, then poured into alkaline water and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. The oil obtained was dissolved in methanol, IPA / HCl and ether were added. The solids were collected by filtration and washed with ether: (2.2 g, 84%), mp: decomposition above 195 ° C.

Preparation of Chiral Intermediates for Examples 17 and 18

Cleavage of 2-amino-7-nitrotetralin

I.

2-Amino-7-nitrotetralin (30 g, 156 mmol) dissolved in 200 ml acetone was added to dibenzoyl-D-tartaric acid (58.7 g 164 mmol) also dissolved in 200 ml acetone. The thick paste obtained was filtered and washed with acetone. This paste was refluxed with 31 water / ethanol / acetonitrile (1: 1: 1) and then filtered hot. The solid obtained by filtration was recrystallized from the above solvent mixture (3x): (5.25 g, 6%) of one isomer was obtained as shown by chiral capillary zone electrophoresis, mp 240-242 ° C.

Similarly, dibenzoyl-L-tartaric acid could be used to obtain the opposite enantiomer using the same solvent system described earlier: (5.3 g, 6%) of one isomer was obtained as shown by chiral capillary zone electrophoresis, bt240 -242 “C.

Example 17 (+) - N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide (a) (+) - 2 - ((Phenyl) carbonyl) ) amino-7-nitrotetralin

To 2-amino-7-nitrotetraline (1.8 g, 9.39 mmol, obtained via dibezoyl-D-tartaric acid) in THF (50 mL) and 10% potassium carbonate solution (100 mL) was added benzoyl chloride (1, 2 ml, 10.1 mmol). After the addition was complete, the reaction mixture was diluted with water to a volume of 250 mL. The precipitated solids were collected by filtration, washed with water and dried in vacuo: (2.8 g, 100%), mp 208-209 ° C, [α] _D + 21.9 ^e (c 0.33 DMSO).

(b) (+) - 2 - ((Phenyl) methyl) amino-7-nitrotetraline hydrochloride

K (+) - 2 - ((phenyl) carbonyl) amino-7-nitrotetralin (2.8 g,

9.4 mmol) in anhydrous THF (100 mL) was added borane-THF (32.8 mL, 1M THF, 32.8 mmol). The mixture was heated to reflux for 5 hours, cooled to 0 ° C and quenched by dropwise addition of 4N HCl. The mixture was then refluxed for 1 hour, evaporated in vacuo and the solid filtered off (washed with water) and dried in vacuo: (2.8 g, 94%), mp 300 ° C, [α] _D +51 1.0 (c 0.33 DMSO).

(c) (+) -2 - ((Phenyl) methyl) amino-7-aminotetraline hydrochloride

To a stirred solution of (+) - 2 - ((phenyl) methyl) amino-7-nitrotetralin (2.8 g, 8.7 mmol) in MeOH (100 mL) was added a catalytic amount of 10% Pd / C. The reaction mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to a solid glassy product homogeneous by TLC; [α] D + 73.3 ° (c 0.87 DMSO).

(d) (+) - N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide

To (+) - 2 - ((phenyl) methyl) amino-7-aminotetraline hydrochloride (2.5 g, 8.7 mmol) in DMF (20 mL) was added S-methyl-2-thiophenethiocarboximide hydroiodide (3.0 g, 10.4 mmol). The mixture was stirred for 4 hours at 45 ° C, then poured into alkaline water and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered and concentrated to an oily product. The oil obtained was dissolved in methanol, IPA / HCl and ether were added. The solids were collected by filtration and washed with ether. Recrystallization from IPA / MeOH / Et ₂ O gave a white solid: (2.5 g, 66%), bt: decomposition above 260 ° C, [α] _D + 44.5 '(c 0.62) DMSO).

Example 18 (-) - N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide (a) (-) - 2 - ((Phenyl) carbonyl) ) amino-7-nitrotetralin

To 2-amino-7-nitrotetralin (1.8 g, 9.39 mmol, obtained via dibenzoyl-L-tartaric acid) in THF (50 mL) and 10% potassium carbonate solution (100 mL) was added benzoyl chloride (1, 2 ml, 10.1 mmol). After the addition was complete, the reaction mixture was diluted with water to a volume of 250 mL. The precipitated solids were collected by filtration, washed with water and dried in vacuo: (2.8 g, 100%), mp 208-209 ° C, [α] D -24.0 ° (c 0.87 DMSO).

(b) (-) - 2 - ((Phenyl) methyl) amino-7-nitrotetraline hydrochloride

To (-) - 2 - ((phenyl) carbonyl) amino-7-nitrotetralin (2.8 g,

9.4 mmol) in anhydrous THF (100 mL) was added borane-THF (32.8 mL, 1M THF, 32.8 mmol). The mixture was heated at reflux for 5 hours, cooled to 0 ° C and quenched by dropwise addition of 4N HCl. The mixture was then refluxed for 1 hour, evaporated in vacuo and the solid filtered off (washed with water) and dried under vacuum: (2.8 g, 94%), bt above 300 ° C, [α] θ -50.4 ⁰ (c 0.39 DMSO).

(c) (-) - 2 - ((Phenyl) methyl) amino-7-aminotetralin hydrochloride

To a stirred solution of (-) - 2 - ((phenyl) methyl) amino-7-nitrotetralin (2.8 g, 8.7 mmol) in MeOH (100 mL) was added a catalytic amount of 10% Pd / C. The reaction mixture was hydrogenated at 50 psi (1 psi = 0.689 x 10 ⁴ Pa) for 1 hour, filtered through celite and concentrated to a solid glassy product which was homogeneous by TLC; [α] D + 74.6 ° (c 0.80 DMSO).

(d) (-) - N - ((2- (Phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide

To (-) - 2 - ((phenyl) methyl) amino-7-aminotetraline hydrochloride (2.5 g, 8.7 mmol) in DMF (20 mL) was added S-methyl-2-thiophenethiocarboximide hydroiodide (3.0 g, 10.4 mmol). The mixture was stirred for 4 hours at 45 ° C, then poured into alkaline water and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water, dried over magnesium sulfate, filtered, and concentrated to an oily product. The obtained oil was dissolved in methanol, IPA / HCl ether was added. The solids were collected by filtration and washed with ether. After recrystallization from a solvent mixture of IPA / MeOH / Et ₂ O, a solid was obtained: (2.7 g, 71%), bt: decomposition above 260 ° C, [α] _D ~ 44.5 (c 0.57)

DMSO).

Example 19

N- (2,3,4,5-Tetrahydro-1H-3-benzazepin-7-yl) -thiophene-2-carboximidamide (a) 2,3,4,5-Tetrahydro-1H-3-benzazepin-7-amine monohydrochloride

To a solution of 2,3,4,5-tetrahydro-7-nitro-1H-3-benzazepine hydrochloride (1.68 g, 7.35 mmol) in ethanol (100 mL) was added 5% palladium on carbon (0.2 g). ), the solution was placed in a Paar hydrogenation apparatus and 45 psi hydrogen (1 psi = 0.689.10 ⁴ Pa) was admitted. After reaching the theoretical consumption of hydrogen (2 h), the catalyst was filtered off and washed with water (25 ml). The filtrate was evaporated. Absolute ethanol was then added and the reaction mixture was evaporated to evaporate all the water to give a solid. This solid was dissolved in hot ethanol (50 mL) and the product was precipitated by addition of ether (75 mL). The solid was collected and air dried to give an off-white solid: (2.43 g, 94%), bt 280-291 ° C.

(b) N- (2,3,4,5-Tetrahydro-1H-3-benzazepin-7-yl) -thiophene-2-carboximidamide

A suspension of 2,3,4,5-tetrahydro-1H-3-benzazepine-7-amine monohydrochloride (0.60 g, 3.0 mmol) and S-methyl-2-thiophenethiocarboximide hydroiodide 1.1 g, 3.8 mmol ) in dimethylformamide (2.0 mL) and isopropanol (2.0 mL) was stirred at room temperature for 20 hours. The solid from this reaction was separated and washed with isopropanol (5 mL) and ethyl acetate (15 mL). The air-dried solid was 1.18 g and was a mixed salt. This solid was dissolved in water, basified and taken up in ethyl acetate. The extract was dried over magnesium sulfate and then evaporated to give the free base as a yellow solid. This product was taken up in isopropanol (30 mL) and hydrogen bromide in isopropanol was added until the solution was acidic. The product was precipitated by the addition of ethyl acetate (35 mL). The precipitated product was collected and dried to give the product dihydrobromide (0.70 g (54%)), m.p.281-3 C.

Example 20

N- (1,2,3,4-Tetrahydroisoquinolin-7-yl) thiophene-2-carboximidamide (a) 1,2,3,4-Tetrahydroisoquinolin-7-amine monohydrochloride

The compound was prepared according to the method of Example 19, step (a). From a 7-nitro-1,2,3,4-tetrahydroisoquinoline hydrochloride (3.00 g, 14.0 mmol) and 5% palladium on carbon (0.3 g) in ethanol (150 mL) a pinkish solid product was isolated. colors (2.43 g (94%)), bt232-4 ° C.

(b) N- (1,2,3,4-Tetrahydroisoquinolin-7-yl) thiophene-2-carboximidamide

The compound was prepared according to the method of Example 19, step (b). From 1,2,3,4-tetrahydroisoquinolin-7-amine monohydrochloride (0.46 g) and S-methyl-2-thiophenethiocarboximide hydroiodide (0.85 g) in isopropanol (2.0 mL) and dimethylformamide (2.0 mL) after work-up of the reaction mixture, the title compound was obtained as the free base (0.60 g (94%)). This material was converted to the bis-oxalate in methanol / ethyl acetate to give an off-white solid (0.59 g (54%)), m.p. 199-200 ’C.

Example 21

N- (2-Benzyl-1,2,3,4-tetrahydroisoquinolin-7-yl) thiophene-2-carboximidamide (a) 2-Benzyl-7-nitro-1,2,3,4-tetrahydroisoquinoline monohydrochloride

To a solution of 7-nitro-1,2,3,4-tetrahydroisoquinoline monohydrochloride (2.50 g, 11.6 mmol) and potassium carbonate (2.0 g) in acetonitrile (100 mL) was added benzyl bromide (2.22). g, 13.0 mmol) in acetonitrile (10 mL). This solution was stirred overnight and the solid was then filtered off. The solvent was distilled off in vacuo and the resulting solid was extracted with a mixture of methylene chloride and water. The dried organic phase (magnesium sulfate) was evaporated and the oily product obtained was taken up in ethanol (50 ml). This solution was acidified with hydrochloric acid in ethanol. 150 ml of ethanol and 50 ml of ether were added to the resulting precipitate. The collected air-dried solid afforded 2-benzyl-7-nitro-1,2,3,4-tetrahydroisoquinoline monohydrochloride as an off-white solid (2.78 g, (79%)), m.p. 256-8 ’C (decomposition).

(b) 2-benzyl-1,2,3,4-tetrahydroisoquinolin-7-amine hydrochloride

The compound was prepared according to the procedure of Example 19, step (a). The product was isolated from 2-benzyl-7-nitro-1,2,3,4-tetrahydroisoquinoline hydrochloride (2.00 g, 6.56 mmol) and 5% palladium on carbon (0.2 g) in ethanol (100 mL). as a yellowish solid (1.05 g (78%)), mp 257-9 ’C (decomposition).

(c) N- (2-benzyl-1,2,3,4-tetrahydroisoquinolin-7-yl) -thiophene-2-carboximidamide

The compound was prepared according to the procedure of Example 19, step (b). From 2-benzyl-1,2,3,4-tetrahydroisoquinoline-7-amine monohydrochloride (0.50 g, 1.8 mmol) and S-methyl-2-thiophenethiocarboximide hydroiodide (0.67 g, 2.3 mmol) in isopropanol (2.0 mL) and dimethylformamide (2.0 mL) was isolated as a yellow solid. This product was converted to oxalate in isopropanol solution, m / e = 348 (M + H) ⁺ .

Example 22

N- (1,2,3,4-tetrahydroisoquinolin-5-yl) thiophene-2-carboximidamide dioxalate

Prepared according to the method ¹ of Example 20, mp 75 C (dec). '

Example 23

N- (1,2,3,4-Tetrahydroisoquinolin-6-yl) thiophene-2-carboximidamide (a) 1,2,3,4-Tetrahydroisoquinolin-6-amine monohydrochloride

Isoquinolin-6-amine solution [Manske, RHF et al., J. Am. Chem. Soc., 72, 4997 (1950)] (4.40 g, 30.5 mmol) and platinum dioxide (300 mg) in a solution of acetic acid (85 mL) and 2.5 M hydrochloric acid (30 mL) were added. was placed in a Paar hydrogen apparatus, and the apparatus was pressurized with hydrogen to 45 psi (1 psi = 0.689.10 ⁴ Pa) for 16 hours. The solvent was distilled off in vacuo and the resulting salt was partitioned between a mixture of aqueous potassium carbonate and 20% isopropanol in methylene chloride. The dried organic phase (magnesium sulfate) was evaporated and the resulting oily product was chromatographed on silica gel, eluting with 2% methanol in chloroform. Obtained as an oily solid (3.08 g (93%)). This product (3.08 g, 20.8 mmol) was taken up into

200 ml of ethanol and 1 equivalent of 0.1000 M hydrochloric acid solution was added. Evaporation of the solvent gave the monohydrochloride as a solid, MS 149 (M + H).

(b) N- (1,2,3,4-Tetrahydroisoquinolin-6-yl) thiophene-2-carboximidamide

The compound was prepared according to the procedure of Example 19, step (b). From 1,2,3,4-tetrahydroisoquinoline-6-amine monohydrochloride (0.90 g, 4.9 mmol) and S-methyl-2-thiophenothiocarboximide hydroiodide (1.80 g, 6.2 mmol) in isopropanol (2) (0 mL) and dimethylformamide (2.0 mL) gave the title compound as the free base (0.74 g (57%)) after work-up and chromatography on silica gel; 170-5 ’C ·.

Example 24

N- (isoquinolin-7-yl) thiophene-2-carboximidamide (a) 7-Nitroisoquinoline

A solution of 7-nitro-3,4-dihydroisoquinoline (3.00 g,

17.0 mmol) and 5% palladium on carbon (3.0 g) in decalin (75 mL) was heated at reflux for 3 hours. After cooling, the solution was filtered and the catalyst was washed with chloroform (200 mL). After distilling off the solvent in vacuo, 7-nitroisoquinoline was obtained as a tan solid. MS: 175 ([M + H]).

(b) Isoquinolin-7-amine

7-Nitroisoquinoline (1.62 g, 9.25 mmol) in ethanol (150 mL) was hydrogenated in a Paar hydrogenation apparatus with a 5% palladium on carbon catalyst (0.2 g) for 3 hours at 50 psi ( 1 psi = 0.689.10 ⁴ Pa). The reaction mixture was then filtered and the solvent was evaporated under reduced pressure. Recrystallization of the solid in ethanol (3 mL) gave isoquinolin-7-amine (0.98 g) as a tan solid. MS 145 (M + H), NMR (CDCl ₃ ) 9.02 (s, 1H), 8.29 (d, 1H), 7.63 (d, 1H), 7.47

- 49 (d, 1Η), 7.13 (dd, Η), 7.03 (d, 1Η), 4.00 (broad, 2H).

(c) N- (isoquinolin-7-yl) thiophene-2-carboximidamide

This solution was extracted

A solution of isoquinolin-7-amine (0.96 g, 6.7 mmol) and S-methyl-2-thiophenethiocarboximide (2.42 g, 8.36 mmol) in isopropanol (4 mL) and DMF (4 mL) was stirred for 18 hours then poured into dilute sodium hydroxide with methylene chloride. The extract obtained was dried over magnesium sulfate and the solvent was evaporated to give an oily product which solidified on standing. The material was purified by silica gel column chromatography (5% methanol in chloroform saturated with ammonia gas) to give 1.31 g of a solid. This material was recrystallized from ethyl acetate (25 mL) to give 1.05 g of the title compound as an off-white product. Mp 177.5-178.5 ’C.

Claims (21)

PATENT CLAIMS A compound of formula I wherein D represents a five-membered heterocyclic aromatic ring containing from 1 to 4 heteroatoms selected from O, N or S, optionally substituted on a carbon atom with halogen, trifluoromethyl, C1-6 alkyl, nitro or cyano, and which is attached to the remainder of the compound of formula I via a carbon atom; A is N (X) or CH (-CH ₂ ) _m -NXY); U represents NH, O or _CH2; W represents (CH _{2) a;} W represents (ΟΗ ₂ ) 2; a and b independently represent integers from 0 to 3, provided that a + b is in the range of 1 to 3; X and Y independently represent hydrogen, alkyl Cl to 6, or - (CH _{2) n} Q, or NXY represents piperidinyl, pyrrolidinyl, morpholinyl or tetrahydroisoquinolinyl; Q represents biphenyl or phenyl optionally substituted with one or more groups selected from alkyl groups C1-6, alkoxy C1-6, perfluoroalkyl C1-6, halogen, nitro- or cyano-; m is an integer from 0 to 5; n is an integer from 0 to 6; or the chain U-V-A-W is defined as above but may be unsaturated; UVAW or chain-chain can be represented by _{-NH-CH2 -CH2} -O- substituted at a carbon atom a group - (CH ₂ ) _m -NXY, wherein m, X and Y are as defined above, and the corresponding pharmaceutically acceptable salts and enantiomers thereof. A compound of formula I according to claim 1 having the structure defined by formula IA: D IA where T is a C ₃ _ ₅ saturated or unsaturated alkylene chain substituted by - (CH _{2) m} NXY; -O (CH _{2) 2} ^- NHsubstituovaný - (CH _{2) m} NXY; or -U- (CH ₂ ) _a - N (X) - (CH ₂ ) _b -; X and Y are independently hydrogen, C 1-6 alkyl, or - (CH ₂ ) _n Q or -NXY is piperidinyl, pyrrolidinyl, morpholinyl or tetrahydroisoquinolinyl; Q is phenyl optionally substituted with C1-6 alkyl, C1-6 alkoxy, trifluoromethyl, halogen, nitro or cyano; and U, m, n, a, b and D are as defined above, but provided that when T is -U- (CH ₂ ) _and -N (X) - (CH ₂ ) _b and X is - (CH _{2) n} is an integer between 0 and 5, and the corresponding pharmaceutically acceptable salts and enantiomers thereof. A compound of formula (I) according to claim 2, wherein T is a ^C 1-5 -saturated or unsaturated alkylene chain substituted with - (CH ₂ ) _m -NXY; or -O (CH ₂ ) ₂ -NHsubstituted - (CH ₂ ) _r -NXY; and X and Y independently represent hydrogen, alkyl C1 to 6 or - (CH _{2) n} Q. A compound of formula 1 according to claim 2 or claim 3 wherein T is a ^C 3-5 saturated or unsaturated alkylene chain substituted with - (CH ₂ ) _m -NXY. A compound of formula I according to any one of claims 1 to 4, wherein m is 0 or 1. A compound of formula 1 according to claim 2, wherein T is -U- (CH ₂ ) _a - N (X) - (CH ₂ ) _b and X is hydrogen, C 1-6 alkyl, or - (CH ₂ ) _n Q. A compound according to claim 2 or claim 6 wherein T represents -U- (CH ₂ ) _and -N (X) - (CH ₂ ) _b - and U represents CH ₂ . A compound according to claim 2, 6 or 7, wherein T is -U- (CH ₂ ) _and -N (X) - (CH ₂ ) _b - and a + b are 1 or 2. A compound according to any one of the preceding claims wherein n represents 0, 1 or 2a X and / or Y represents - (CH ₂ ) _n Q. A compound according to any one of the preceding claims wherein X and / or Y is - (CH ₂ ) _n Q and Q is phenyl optionally substituted with C 1-6 alkyl or halogen. A compound of formula I according to any one of the preceding claims wherein D represents a five membered heterocyclic ring containing one heteroatom selected from O, N or S, optionally substituted on a carbon atom by halogen. A compound of formula I according to claim 11, wherein D represents thienyl, pyrrolyl or furyl. A compound of formula I according to claim 12, wherein D represents 2-thienyl. 14. A compound of formula I which is: N - ((2- (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide N - ((2- (phenylmethyl) amino) -1,2,3,4-tetrahydronaphth-7-yl) -2- thiophenecarboximidami; N - ((2-amino) -1,2,3,4-tetrahydro-7-yl) -2-thiophenecarboximidamide; N - ((1-amino) -1,2,3,4-tetrahydronaphth-7-yl) -2-thiophenecarboximidamide; N - ((2-amino) indan-5-yl) -2-thiophenecarboximidamide; N - ((2- (methyl) (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide; N - ((1-amino) indan-6-yl) -2-thiophenecarboximidamide; N - ((1- (phenylmethyl) amino) indan-6-yl) -2-thiophenecarboximidamide N - ((2 - ((3-chlorophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxaraidine; N - ((2 - ((2-methylphenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - ((3-methylphenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - ((4-methylphenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2- (Ethyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - (((4-phenyl) phenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - (((4-hexyl) phenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - ((3-bromophenyl) methyl) amino) indan-5-yl) -2-thiophenecarboxamidine; N - ((2 - ((3-chlorophenyl) methyl) amino) -l, 2,3,4-tetrahydro-7-yl) -2-thiophenecarboximidamide; N - ((2- (phenylmethyl) (methyl) amino) -1,2,3,4-tetrahydro-7-yl) -2-thiophenecarboximidamide; N - ((1- (phenylmethyl) amino) -1,2,3,4-tetrahydro-7-yl) -2-thiophenecarboximidamide; N - ((1- (phenylmethyl) amino) indan-5-yl) -2-thiophenecarboximidamide; N - ((i- (phenylmethyl) amino) -l, 2,3,4-tetrahydro-6-yl) -254 thiophenecarboximidamide; N - ((2- (phenylmethyl) amino) -1,2,3,4-tetrahydro-7-yl) -2-furancarboximidamide; N- (2,3,4,5-tetrahydro-3-benzazepin-7-yl) thiophene-2-carboximidamide; N- (1,2,3,4-tetrahydroisoquinolin-7-yl) thiophene-2-carboximidamide; N- (2-benzyl-l, 2,3,4-tetrahydroisoquinolin-7-yl) thiophene-2-carboximidamide; N- (1,2,3,4-tetrahydroisoquinolin-5-yl) thiophene-2-carboximidamide; N- (1,2,3,4-tetrahydroisoquinolin-6-yl) thiophene-2-carboximidamide; N- (isoquinolin-7-yl) thiophene-2-carboximidamide; or a pharmaceutically acceptable salt or enantiomer thereof. A compound according to any preceding claim for use as a pharmaceutical preparation. A pharmaceutical composition comprising a compound as claimed in any one of claims 1 to 14 together with a pharmaceutically acceptable diluent or carrier. Use of a compound according to any one of claims 1 to 14 for the manufacture of a medicament, for the treatment or prophylaxis of diseases or conditions to which the synthesis or over-synthesis of NO-synthase contributes. Use of a compound according to any one of claims 1 to 14 for the manufacture of a medicament for the treatment or prophylaxis of neurodegenerative diseases, migraine or the prevention and alteration of tolerance to opiates and diazepines or for the treatment of drug addictions. A method for the treatment or prophylaxis of diseases or conditions contributing to the synthesis or over-synthesis of NO synthase, comprising administering a therapeutically effective amount of a compound of formula I as defined in any one of claims 1 to 14 to persons suffering from or susceptible to this state. A method for the treatment or prophylaxis of neurodegenerative diseases or migraine or for the prevention and alteration of tolerance to opiates and diazepines or for the treatment of drug addiction, comprising administering a therapeutically effective amount of a compound of formula I as claimed in any one of claims 1 to 14 to sufferers. .these diseases or conditions. 21. A process for the preparation of a compound of formula (I), comprising: (a) preparing a compound of formula I by reaction of a corresponding compound of formula II wherein U, V, A and W are as defined in claim 1 with a compound of formula III D Wherein D is as defined in claim 1 and L is a leaving group; (b) preparing a compound of formula I by reacting the corresponding compound of formula IV I A. nh ₂ . ha Wherein U, V, A and W are as defined in claim 1 and HA is an acid, with a compound of formula V Where D is as defined in claim 1; (c) a compound of formula I wherein A represents N (X) and X represents alkyl C1 to 6 or - (CH _{2) n} Q by reacting a corresponding compound of formula I wherein X is hydrogen with a compound of formula VI R ⁹ -L VI wherein R ⁹ is C 1-6 alkyl or - (CH ₂ ) _n Q and L is a leaving group; (d) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY) and at least one X or Y is C 1-6 alkyl or - (CH ₂ ) _n Q <reaction of the corresponding compound of formula I, wherein one or both of X and Y are hydrogen, with a compound of Formula VI; (e) preparing a compound of formula I, ^one in which A represents CH (- _{(CH2) m-} NXY) and m represents an integer of 1 to 5, by reduction of a corresponding compound of formula VII XYNCO (CH ₂ ) mi-- VII wherein U, V, W, X, Y and D are as defined in claim 1; (f) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY) and both X and Y are hydrogen, by reducing the corresponding compound of formula VIII O ₂ N- (CH ₂ ) _m VID wherein U, V, W, m and D are as defined in claim 1; (g) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _n -NXY), X is hydrogen and _m is an integer from 1 to 5, by reducing the corresponding compound of formula IX N-Y-I-ChiChan IX wherein U, V, W, D and Y are as defined in claim 1; (h) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY), X or Y is hydrogen and the other is - (CH _{2) n} Q ⁱⁿ which n represents an integer of from 1 to 6 by reduction of a corresponding compound of formula X Q (CH ₂ ) n.iCONH (CH ₂ ) r - ^H % D> = NH wherein Q, m, U, V, W and D are as defined in claim 1; (i) preparing a compound of formula I wherein A is CH (- (CH ₂ ) _m -NXY), X or Y is hydrogen and the other is - (CH ₂ ) _n Q wherein n is an integer from 1 to 6, by reducing the corresponding compound of formula XI Q (CH ₂₎ n4CH = N- _(CH2) m ^HC) ^ NH XI wherein Q, m, U, V, W and D are as defined in claim 1; or (j) preparing a compound of formula I wherein A is CH (-NXY) and X is hydrogen, by reducing the corresponding compound of formula XII XII wherein U, V, W, D and Y are as defined in claim 1; and, if desired or necessary, converting the resulting compound of formula I or another salt thereof into a pharmaceutically acceptable salt thereof, or vice versa.