2-(AMINOALKYLTHIO)METHYL-1,4-DIHYDROPYRIDINE, A METHOD FOR THE PREPARATION THEREOF AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

Abstract: Compounds of formula I wherein P and P₁ groups represent hydrogen, alkyl, hetero­ alkyl, aryl or aralkyl groups, optionally substituted or forming a ring, n is an integer between 0 and 2, m is an integer from l to 3, R₁ and R₃, that may be the same or different, represent alcoxycarbonyl groups, CN, NO₂ or acyl groups, whereas R₂ is an aryl group or an heteroaryl group that may be optionally substituted, are useful as active principles of pharmaceutical compositions.

Description: Object of the present invention are 2-(formylaminoalkylthio)-methyl-l,4-dihydropyridine, a method for the preparation thereof and pharmaceutical compositions containing them. The compounds object of the present invention have the following general formula I wherein: R₁ represents: . a COCH₃, COC₆H₅, CN or NO₂ group; . a COORa group, wherein Ra represent hydrogen, C₁-C₆ straight or branched alkyl, optionally bearing one or more C₁-C₆ alkoxy groups and/or secondary amino groups of formula -NR₄R₅ wherein R₄ and R₅, that may be the same or different, represent C₁-C₆ alkyl, phenyl, benzyl or taken, together with the nitrogen atom, form a five or six membered ring optionally containing other atoms; R₂ is a phenyl ring unsubstituted or substituted with one or more C₁-C₆ alkyl, halogen, nitro, cyano, C₁-C₆ alkoxycarbonyl, C₁-C₆ alkylthio, C₁-C₆ alkylsulfynyl; pentafluorophenyl; α- or β-naphtyl; a five- or six-membered heterocyclic ring; α-benzo[2.3-b]-l.4- dioxan-α-yl; α-benzofuroxanyl; R₃ represents a COORa group wherein Ra is as above defined; P is selected from the group consisting of hydrogen, -(CH₂) p -W and C₁-C₈ linear or branched alkyl; N-P₁ is the residue of a primary or secondary amino group wherein P₁ is selected from the group consisting of hydrogen, C₁-C₆ lower linear or branched alkyl group and -(CH₂)p-W; P, taken together with P₁ and the nitrogen atom to which P₁ is linked, may form a pyrrolidine or a piperidine ring; W is selected from the group consisting of hydroxymethyl, formyloxymethyl, CO₂R wherein R is hydrogen or C₁-C₄ lower alkyl, CN, saturated or unsaturated heterocyclic ring, C₃-C₇ cycloalkyl ring, phenyl ring optionally substituted by one or more halogens or C₁-C₃-alkoxy groups; m is an integer from l to 3; n is zero or an integer from l to 2; p is zero or an integer from l to 3. Also the pharmaceutically acceptable salts as well as the optical antipodes, i.e. the enantiomers, the possible geometric isomers, diastereoisomers and mixtures thereof are included in the scope of the present invention. The alkyl, alkenyl, alkoxy and alkanoyloxy groups are branched or straight chain groups A halo-C₁-C₆ alkyl group is preferably trihalo -C₁-C₆ alkyl, in particular trifluoromethyl. A halo -C₁-C₄ alkoxy group is preferably -OCHF₂. A C₁-C₆ alkyl group is preferably methyl, ethyl, isopropyl or t-butyl. An aryl group is preferably phenyl. A C₃-C₅ alkenyl group is preferably allyl. A C₃-C₅ alkynyl group is preferably propargyl. A C₃-C₇ cycloaliphatic group is preferably cyclopentyl, cyclohexyl or cycloheptyl. A monoalkyl amino group is preferably a methyl-, ethyl-, isopropyl- or benzyl-amino group. A C₁-C₃ alkoxy is preferably methoxy or isopropoxy. A C₁-C₃ alkylthio is preferably methylthio or isopropylthio. A C₁-C₄ alkoxycarbonyl is preferably methoxy-, ethoxy- or ter-butoxy-carbonyl group. When R₂ is a five- or six- membered heterocyclic ring, it is preferably pyridyl, furanyl or thienyl. The non toxic salts, that are pharmaceutically acceptable, include the hydrochlorides, hydrobromides, hydroiodides, (lower)alkylsulfates, (lower)alkyl and aryl sulfonates, phosphates, sulfates, maleates, fumarates, succinates, tartrates, citrates, and others commonly used in the art. The salts obtained through the variation of the acid used in some cases have special advantage due to increased stability, increased solubility, decreased solubility, ease of crystallization, lack of objectionable taste, etc., but these are all subsidiary to the main physiological action of the free base, which is independent on the character of the acid used in the preparation of the salt. Specific examples of preferred compounds of the invention are: a 4-(3-nitrophenyl), (3-chlorophenyl), (3-cyanophenyl), (3-methoxyphenyl), (4-fluorophenyl), (3-methylthiophenyl), (α-benzo[2,3-b]- l,4-dioxan-α-yl), (2-fluoro-5-methylthiophenyl)- 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-6-methyl- l,4-dihydropyridine. a diastereoisomer of 2-(2-formylamino-2-phenylethylthio) methyl-3-ethoxycarbonyl-5-methoxy carbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine. a diastereoisomer of 2-(N-formyl-pyrrolidin-2-yl methylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4--(m-nitrophenyl)-6-methyl-l.4-dihydropyridine. 2-(2-formylaminoethylthio) methyl-3-ethoxycarbonyl-5--(2-N,N-dimethylamino)ethoxycarbonyl-4-(m-chlorophenyl)-6-methyl-l,4-dihydropyridine. 2-(2-formylaminoethylthio)methyl-3-methoxyethoxycarbonyl-5-methoxycarbonyl-4-(3-methylthiophenyl)-6-methyl-l,4-dihydropyridine. 2-[2-N-(2-cyanoethyl)-N-formylaminoethylthio]methyl--3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)--6-methyl-l,4-dihydropyridine. 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5--(2-N-methyl-N-benzylamino)ethoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine. The compounds of the invention are obtained by a process comprising a) forming a thioether bond by reaction of a compound of general formula (II) with a compound of formula III wherein: R₁, R₂, R₃, m, P, N-P₁ are as above defined; either Y or Y′ represents a thiol or a masked thiol group such as thio C₂-C₁₂ alkanoyl ester or a thiouronium salt -S-(C=NR₆)NR₇R₈ (+) y₂ (-) the other being a known leaving group such as chlorine, bromine, iodine, trifluoromethane sulphonate, an alkyl- or an arylsulphonate; R₆, R₇, R₈, which are the same or different, are hydrogen or a C₁-C₄ alkyl group and y₂ (-) is a pharmaceutically acceptable anion; b) cyclizing a compound of formula IV with an alkylidene compound of formula V wherein: R₁, R₂, R₃, P, NP₁, m, are as above defined. c) cyclizing a compound of formula VI with an alkylidene compound of formula VII wherein R₂, R₃, P, NP₁ and m are as above defined and R₁, is preferably CO₂Ra, CN, COCH₃, COC₆H₅. d) formylating a compound of formula VIII wherein R₁, R₂, R₃, P, P₁, and n are as above defined. The above reactions a) - d) give compounds of formula I which may be optionally subjected to further processes such as oxidation, salification, separation of isomers etc. The thioeterification reaction between a compound of formula II and a compound of formula III may be carried out, as known in the art, using almost equimolar amounts of reagents in an inert solvent at a temperature ranging form -20° to +60°C, in the presence of a base. Suitable solvents may be chosen in the group of alcohols, amides, linear or cyclic ethers, ketones esters, halogenated hydrocarbon, whereas the base may be an alkali or earth-alkali metal hydroxide, carbonates, bicarbonates, alcoholates, hydrides, amides or organic bases such us triethylamine, pyridine etc. The selective oxidation of the thioethereal bond of the compounds of formula I where n=O to give the compounds of formula I wherein n is l or 2 is carried out in an inert solvent such as ethyl acetate, ethyl formate, dichloromethane, l,2-dichloroethane, chloroform or mixtures thereof, by reaction with one or more molar equivalents of a peracid such as perbenzoic, m-chloro-perbenzoic, periodic, monoperftalic, peracetic, performic or peroxytrifluoroacetic acid, working in a temperature ranging from -30°C to room temperature. Preferably, the reaction is carried out at above 0°C. Compounds of formula I wherein n=l are obtained using l mol. equiv. of peracids, while using 2 or mor mol. equiv. of peracids, compounds with n=2 are obtained. The cyclization of an enaminoderivative of formula IV and VI with an ethylene derivative of formula V and VII respectively is the well-known Hantzsch reaction, described by F. Brody and P.R. Ruby in "Pyridines and its derivatives", part I, pages 355-533, A. Weissbenger -Interscience, New York l960. The formylation reaction of a compound of general formula VIII is also carried out using well-known technique, for instance by treatment in an inert solvent such as dimethyl formamide or dimethyl acetamide, with a 5-l0 molar excess of formic acid and heating the mixture for l 8 h at a temperature ranging from 70 to l20- °C. Alternatively, an amino compound of formula VIII is reacted in an inert dry solvent such as tetrahydrofuran, benzene, ethylacetate, dimethylformamide or their mixtures with at least a 0.l molar excess of N-formyl-imidazole; preferred reaction conditions are those from the room temperature to 5°C for a time from l0 minutes to l-2 hours. Alternatively, a salt of a compound of formula VIII (hydrobromide, nitrate, sulphate or emisulphate) is reacted with an excess of formamide at a temperature ranging from 60 to 90°C for a time from few minutes to 3 hours. The optical resolution processes are preferably carried out on compounds of formula I wherein n is 0 or 2, whereas chiral compounds wherein n is l are preferably obtained by oxydation of the antipodes wherein n is zero. The optional resolution process is carried out on diastereoisomeric derivatives of compounds I. For examples, diastereoisomeric salts of compounds I with optically pure acids or bases may be preparared by known salification methods and optionally subjected to optical resolution. When Ra contains an amino group, optically pure acids are used while when Ra is hydrogen, optically active bases are used, to give, after the resolution process, optically pure acids of formula I, which may be optionally esterified by known methods. On the other hand, racemic acids of formula I may be esterified with optically pure alcohols and the obtained diastereoisomeric alcohols may optionally be subjected to resolution by crystallization or chromatographic methods. The obtained optically pure esters may be transformed by known methods into acids or esters I. The compounds of formula II wherein Y is a halogen, have been described in the European Application n. 86110295.2 and the compounds of general formula IV wherein y is a thiol and/or a masked thiol function are described in PCT/EP86/00445. The compounds of formula III, if not already known may be easily prepared from commercialy available compounds, using well-known, safe and unexpensive methods. A general source of compounds III are aminoalcohols IX wherein m, P, P₁ are as above defined; said alcohols are commercially available or are prepared by reduction of the corresponding amino-carboxy esters or lactames. Compounds of general formula IX are easily converted into compounds III by known reactions such as formylation and transformation of the alcoholic group into a thiol. Compounds of general formula V are prepared from compounds of general formula X (P, P₁, X, m and Ra as above defined) by Knoevenagel condensation with aldehyde of formula XI R₂CHO (XI) (R₂ as above defined) while compounds of formula VII are prepared from compounds of formula X by reaction with ammonia or ammonium salt. Compounds of formula X are prepared, for example, via thioetherification reaction of compounds of formula III with 4-chloro-3-oxo-butanoic acid Compounds of general formula IV and XI are known or easily available. Some formylamino alkyl thiomethyl derivatives of the invention, such as 2-formylamino ethyl thio-4-(m-nitrophenyl)-3-carboethoxy-5-carbomethoxy-6-methyl-l,4-dihydropyridine show peculiar properties such as low oral acute toxicity and high tolerability in some susceptible experimental animals, e.g. dogs, combined with pronounced and long lasting antihypertensive activity at very low doses (for ex. 0.2 mg/kg/os) when tested orally in conscious SH rats once daily. The antihypertensive effect is dose related in the investigated dose range, for example from 0.05 to 0.8 mg/kg. The maximum hypotensive effect, proportional to the administered dose, takes place 5-7 hours after the administration and the blood pressure is maintained at the decreased level for further 4-5 hours, at least. The gradual onset of the antihypertensive effect does not seem to be coupled with reflex tachicardia, which is often observed after treatment with other antihypertensive agents such as, for example, hydralazine and many dihydropyridines, in the same experimental models. On the contrary, no substantial modifications of the mean B.P. and heart rate are observed after one day oral administration of the same compounds to normotensive conscious rats at the same dose range. After a two weeks treatment consisting of one daily oral administration using a dose range from 0.l to 0.8 mg/kg, a gradual decrease of mean B.P. also dose-related is observed in the treated conscious SH-rats. Dosages such as 0.2-0.4 mg/Kg are sufficient to stabilize gradually the mean B.P. to lower level for a 24 hrs period. The drop-out of the pharmacological treatment is not combined with an acute hypertensive rebound effect but in the following 2-3 days the mean blood pressure gradually rises to the initial values. A similar pharmacological profile is shared by many 2-amino alkyl thio-methyl-l,4-dihydropyridines of formula III. In general they turn out to be effective antihypertensive agents when orally tested in consciuous SH rats and well-tolerated in subchronic toxicological studies carried out in male and female normotensive rats, independently from the fact that many of these substances show low LD₅₀ in mice by the oral or intraperitoneal route. Representative results of acute toxicity studies are reported in table l; the same substances tested orally in conscious SH rats (200-250 g/body weight) show at l.6 mg/kg a maximum decrease of the mean blood pressure of about 30-80 mmg, with the exclusion of compounds l, 4, 2l, 20, 3l (- BP ranging from l0 to 25 mmHg); 2, 5, 6, 7, l0, ll, l2 (- BP higher than 85 mmHg), whereas the compound l6 d is practically inactive at 3.2 mg/kg. However, male and female beagle dogs orally treated with l.5 mg/kg/day (bolus administration) of the compounds ll and l5 died after three days treatment. Acylation of the amino group of a compound of formula VIII, (for ex. the 2-acetyl amino or 2-benzoylamino derivatives of compounds ll, l5) reduces acute toxicities in mice but in general the long lasting antihypertensive action in SH rats is lost and the compounds require almost two administrations daily in order to stabilize the mean blood pressure values to lower levels for a 24 hours period. Surprisingly, the 2-formyl amino-alkyl-thio methyl-l,4-dihydropyridines of the present invention do not only show long lasting antihypertensive effect and lower acute toxicities (for ex. 2-formyl-amino-ethyl-thio methyl-4-(m-nitrophenyl) 3-carboethoxy-5-carboethoxy-4-m-nitrophenyl-6-methyl-l,4-dihydropyridine shows LD₅₀ in mice of 200 and l50 mg/kg orally and i.p. respectively) but are well-tolerated in dogs. Male and female beagle dogs orally treated with the latter substance of the invention at dosages as high as 5-l2.5 mg/kg/day (bolus administration) do not die after two weeks treatment. After N-formylation of the side chain amino function of the compound of formula VIII of table I, a 5 to l2 fold acute toxicity reduction in mice is observed. For example, N-formylation of compound 8, increases LD₅₀ in mice (oral) from 8 mg/kg to 90 mg/kg. Moreover, the ability of the compounds of the invention to inhibit "in vitro" the contractile activity induced by increasing concentration of calcium ions in K⁺-depolarized rat aorta strips was investigated according to the technique of T. Godfraind et al. (Arch. Int. Pharmacodyn. l72 , 235, l968). Representative results of these studies with compound ll and its N-acetyl and N-formyl derivative are reported in Table 2. The reported results further confirm that better inhibiting activities (ID₅₀ ranging from l0⁻⁷ to l0⁻¹⁰) may be observed after longer incubation times with the investigated tissue preparations than after shorter ones (ID₅₀ ranging from l0⁻⁵ to l0⁻⁹) whereas the behaviour of the standard compound nifedipine is not related to the incubation time. The shorter lasting antihypertensive effect of the N-acetyl compound and the long lasting effects of the compounds ll and of its N-formyl derivative does not seem to be correlated with "in vitro" experimental results related to the supposed Ca-antagonist potency and capability of relaxing smooth muscles, phenomenon which cannot be easily explained on the basis of the present knowledge. Independently from this aspect, the particular antihypertensive effect, its gradual onset, the long lasting action combined with lower acute toxicity and with increased tolerability in dogs provide evidence that the compounds of the invention are useful in human and veterinary therapy for the treatment of hypertensive situations of different origin, and for the treatment and prevention of cardiovascular and coronary diseases. In order to attain the desired effects in human and veterinary therapy, the compounds of the invention may be administered parenterally, for example by intravenous, hypodermic or intramuscular injection, by infusion, rectally or orally. The compounds may also be administered in pure form or in the form of a pharmaceutical composition. The formulation of suitable pharmaceutical compositions may be prepared according to techniques well-known in the art, such as the ones described in "Remington's Pharmaceutical Sciences Handbook", Hack Publishing Co., U.S.A. When the compounds of the invention are used as antihypertensive drugs, the dosage will vary according to the seriousness of the hypertension and to the administration route. The amount of the active principle administered may vary, anyhow, between lmcg/kg/die and l mg/kg/die, preferably between 5 mcg/die and 0.l mg/kg/die, by the oral route and from 0.l mcg/kg/die to 0.5 mg/kg/die, preferably from 0.5 mcg/kg/die to 0.2 mg/kg/die, by the parenteral route. A dosage for oral administration may contain, for example, between 50 mcg to l0 mg of active principle. The compounds of the invention may be administered even once a day, however more spaced and/or repeated administrations may be, at least in some cases, more suitable and may vary according to the conditions of the patient and to the administration route or to the dosage. By oral administration the compound may be formulated in solid or liquid preparations, such as capsules, pills, tablets, powders, solutions, suspensions or emulsions. By parenteral administration the compound may be administered in injectable formulations, dissolved or suspended in physiologically acceptable diluents, with a vehicle that may be a sterile liquid such as water or an oil, with or without the addition of other excipients. The compounds may also be administered per rectal route, in the form of suppositories, mixed with the conventional vehicles. The preferred administration route of the compounds of the invention is the oral route. The invention is illustrated by the following non limitative examples, wherein the abbreviations EtOH, DME, THF, Et₂O, AcOEt, AcOH refer to ethanol, dimethoxyethane, methanol, tetrahydrofuran, ethyl ether, ethyl acetate, acetic acid respectively. Preparation l A stirred solution of cysteamine hydrochloride (g 20) in formamide (ml 20) in heated at 75-80°C for 2 hours. After cooling at room temperature, the NH₄Cl precipitate is filtered off and washed with a little formamide. The solution of N-formyl cysteamine (about l8 g) in formamide is then diluted with EtOH (l60 ml), cooled at 0°C, and in nitrogen atmosphere is treated under stirring with 20% aqueous NaOH (l70 ml) and with a solution of ethyl 4-chloro-3-oxo-butanoate (28,4 g) in EtOH (20 ml). After 30 minutes the mixture is poured into water (2000 ml) and extracted with AcOEt (3×200 ml). The combined extracts are washed with a saturated solution of NaH₂PO₄ (3×50 ml) H₂O (3×l00 ml), dried on Na₂SO₄ and evaporated to dryness in vacuum to give 40.5 g of ethyl 4-(2-formylaminoethylthio) 3-oxo-butanoate as an oil; lH-NMR (CDCl₃), δ(TMS):l.l0-l.30 (3H, t); 2.20-2.60 (4 H, m); 3.40-4.l0 (6H, m); 6.60 (lH, m); 8.l0 (lH, S). Using, in the same procedure, the methyl and the methoxyethyl 4-chloro-3-oxobutanoates, the corresponding methyl and methoxyethyl 4-(2-formylaminoethylthio)-3-oxo-butanoates are prepared. Preparation 2 Acetic acid is added to a solution of ethyl 4-(2-formylamino-ethylthio)-3-oxo-butanoate (l2.5 g) in MeOH (ml l20), previously saturated with ammonia at 0°C and cooled at 0°C, up to pH 4.5. The mixture is refluxed for 2 hours and the excess solvent is evaporated in vacuum, to give a syrup from which a solid material is separated by treatment with AcOEt. After filtration, the organic phase is washed with water (8×l0 ml), and with a saturated aqueous solution of NaHCO₃ (3×l0ml), dried (Na₂SO₄) and concentrated in vacuum to give ll,5 g of ethyl 3-amino-4 (2-formylamino ethylthio)crotonate as a yellow oil. lH-NMR:(CDCl₃), δ(TMS):l.l0-l.20(3H,t); 2.20-2.60 (4H, m); 3.80-4.l0 (4H, m); 5.20-5.40 (2H, m); 5.70 (lH, m); 6.60 (lH, m); 8.l0 (lH, s). Methyl and methoxyethyl 3-amino-4-(2-formylaminoethylthio) crotonate are likewise prepared. Preparation 3 A solution of 3 chlorobenzaldeyde (l0 g), ethyl 4-(2-formylaminoethylthio)-3-oxo-butanoate (l6,7 g), AcOH (2 ml) and piperidine (0,6 ml) in benzene (l20 ml) is refluxed in a Dean-Stark apparatus for 6 hours. After cooling at room temperature, the mixture is washed with H₂O (3×20 ml), with a saturated solution of NaHCO₃ (3xl0 ml), with a solution of H₂SO₄ (2N, 3×l0 ml) and again with H₂O (3×30 ml). The organic phase is dried (Na₂SO₄) and concentrated in vacuum to give l6 g of ethyl 2-Z, E-(3-chlorophenylmethylene)-4-(2-formylaminoethylthio)-3-oxo-butanoate, as an oil. lH-NMR(CDCl₃), δ (TMS), l.l-l,2.(3H, t); 2.2-2.4 (2H,t); 2.7-3.0 (2H, m); 8.9-4.2 (4H, m); 6.8-7.9 (7H, m). EXAMPLE l A solution of ethyl 3-amino-4-(2-formylamino ethylthio) crotonate (5.3 g) and 3-Z, E-(m-nitrophenylmethylene) - 2,4-pentanedione (4.9 g; from Knoevenagel condensation of 3-nitrobenzaldeyde with 2,4-pentanedione) in EtOH (l00 ml) is refluxed for 4 hours, it is cooled at 0°C and acidified (pH l:2) with few drops of EtOH saturated with gaseous HCl. After l5 minutes the solvent is evaporated at reduced pressure, the residue dissolved in AcOEt (80 ml), washed with a saturated solution of NaHCO₃ (3×l5 ml), with water (3×30 ml), dried (Na₂SO₄) and concentrated. The residue is purified by chromatography on SiO₂ (300 g, eluent AcOEt/MeOH 80/20) to give 4.9 g of 2-(2-formylaminoethylthio) methyl-3-ethoxycarbonyl-4-(m-nitrophenyl)5-acetyl-6-methyl-l,4-dihydropyridine, m.p. l40-l42°C (EtOH). EXAMPLE 2 A solution of ethyl 3-amino-4-(2-formylaminoethylthio) crotonate (l g) and methyl 2-Z,E- (2 nitro-5-methylthiophenylmethylene)-3-oxobutanoate (0,95 g,m.p. 69-7, obtained by Knoevenagel condensation of methyl acetoacetate and 2-nitro-5 methylthiobenzaldehyde), in EtOH (ml l0) is refluxed for 24 hours, then it is evaporated to dryness. The residue is dissolved in Et₂O (ml 30), washed with HCl (2N, 3×5 ml), H₂O (3×l0 ml), dried and concentrated in vacuum. After column chromatography on SiO₂ 30 g, eluent Et₂O/AcOEt 60/40) l.l g of 2-(2-formylamino ethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(2-nitro-5-methylthiophenyl)-6-methyl-l,4 dihydropyridine, m.p. l48-l50° (Et₂O) are obtained. Using the procedure of example l or 2 the following compounds are prepared: -2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(2, 3-dichlorophenyl)-6-methyl-l,4-dihydropyridine, m.p. l22-l24°C. -2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-cyano-4-phenyl-6-methyl-l,4-dihydropyridine -2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-cyano-4-(3-methoxyphenyl)-6-methyl-l,4-dihydropyridine -2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-benzoyl-4-phenyl-6-methyl-l,4-dihydropyridine -2-(2-formylaminoethylthio)methyl-3-methoxycarbonyl-5-methoxycarbonyl-4-(2-fluoro-3-methythio-phenyl)-6-methyl-l,4-dihydropyridine 2-(2-formylaminoethylthio)methyl-3-methoxycarbonyl-5-methoxycarbonyl-4-(2-methylthiophenyl)-6-methyl-l,4-dihydropyridine. Example 3 A solution of ethyl 3-amino-4-(2-formyl-aminoethylthio)crotonate (7 g) and methyl 2-E,Z-(3-nitro-phenylmethylen)-3-oxobutanoate (6 g) in ETOH (70 ml) is refluxed for l8 hours, then it is concentrated in vacuum. The residue is dissolved in AcOEt (l00 ml), washed with HCl (2N, 3 × 30 ml), H₂O (3 × 50 ml), dried on Na₂SO₄ concentrated in vacuum and purified by column chromatogra phy on SiO₂ (300 g, eluent AcOEt-hexane 90/l0), obtaining 8 g of 2-(2-formylaminoethylthio)-methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l.4-dihydropyridine, mp. l09-ll°C. Example 4 A solution of ethyl 2-Z,E-(3-chlorophenylmethylene)-4-(2-formylaminoethylthio)-3-oxo-butanoate (4 g) and methyl 3-aminocrotonate (l.2 g) in EtOH (40 ml) is refluxed for 3 hours, the mixture cooled at room temperature acidified to pH l:2 with a few drops of EtOH saturated with gaseous Hcl and, after 20 minutes, evaporated to dryness; a solution of the residue in AcOEt (60 ml) is washed with a saturated solution of NaHCO₃ (3 × l0 ml), H₂O (3 × 20 ml), dried (Na₂SO₄) and concentrated in vacuum. After chromatography on silica gel (l50 g; eluent: AcOEt) 4.3 g of 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-4-(3-chlorophenyl)-5-methoxycarbonyl-6-methyl-l.4-dihydropyridina are obtained as a foam. ¹H-NMR (CDCl₃): δ (TMS) = l.l0-l.25 (3H, t); 2.20-2.80 (5H, m); 3.20-3.70 (5H, m); 3.80-4.20 (4H, m); 5.00 (lH, s); 6.50 (lH, m) 7.00-7.20 (5H, m); 8.l0 (lH, s). Using the above described procedure, the following compounds are prepared - 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-cyano-4-(m-chlorophenyl)-6-methyl-l.4-dihydropyridine - 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-(2-N,N-dimethylamino)ethoxycarbonyl-4-(m-chlorophenyl)-6-methyl-l.4-dihydropyridine - 2-(2-formylaminoethyltio)methyl-3-ethoxycarbonyl-5-isopro poxycarbonyl-4(m-chlorophenyl)-6-methyl-l.4-dihydropyridine - 2-(2-formylamonoethylthio)methyl-3-ethoxycarbonyl-5-t-butoxycarbonyl-4(m-chlorophenyl)-6-methyl-l.4-dihydropyridine - 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l.l4-dihydropyridine, mp. l09-ll°C. Example 5 In nitrogen atmosphere, a stirred mixture of 2-mercaptomethyl-3-ethoxycarbonyl-5-methoxycarbonyl-4(m-nitrophenyl)-6-methyl-l.4-dihydropyridine (5 g), K₂CO₃ (2 g) and (S)-2-formylamino-2-phenyl-ethanolmethanesulfonate (3 g from S(+)-N-formylphenylglycin) in DMAF (50 ml) is heated for l2 hours at 50°C. The cooled mixture is poured in ice-water (500 ml) and extracted with AcOEt (30 ml × 3). After usual work-up, the combined organic extracted are evaporated to dryness, to give a diasteroisomer mixture of 4(S)2′(S) and 4(R)2′(S)-2-(2′-formylamino-2′-phenylethylthic)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l.4-dihydropyridine, which are separated by HPL-chromatography on SiO₂ (eluent AcOEt/hexane). The more polar diasteroisomer (l.7 g) is a foam, the less polar is obtained as crystalline compound (2.2 g, m.p. 66-70°C, C₂₇H₂₉N₃O₇S.2/₃Et₂O). Using the above described procedure the following compounds are prepared: - 2-(2-formylamino-3-phenylpropylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l.4-dihydropyridine; - 2-(2-formylaminopropylthio)methyl-3.5-di-ethoxycarbonyl-4-(m-trifluoromethylphenyl)-6-methyl-l.4-dihydropyridine; - 2-/2-formylamino-3-(4-imidazolyl)propylthio/methyl-3-methoxycarbonyl-5-isopropoxycarbonyl-4-(m-cyanophenyl)-6-methyl-l.4-dihydropyridine - 2′-R-2-(2′-formylamino-2'-phenylethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l.4-dihydropyridine - 2-(2-formylamincethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl-6-methyl-l.4-dihydropyridine, m.p.l09-lll°C. Example 6 A solution of 2-chloromethyl-3-ethoxycarbonyl-4-(o-methylthiophenyl)-5-methoxycarbonyl-6-methyl-l.4-dihydro pyridine (2.l g) in EtOH (l0 ml) is added dropwise at 0°C to a solution of N-n-butyl-N-(2-acetylthioethyl)formamide (l.2 g) and NaOH (20% water solution, l.2 g). After 3 hours at 0°C, the reaction mixture is warmed at room temperature and stirred for 30 minutes, then it is concentrated in vacuum. After usual work-up, and column chromatography on SiO₂ (80g; eluent AcOEt/Et₂O 70/90) l.8 g of 2-/2-(N-formyl-N-butylamino)ethylthio/methyl-3-ethoxycarbonyl-4-(o-methylthiophenyl)-5-methoxycarbonyl-6-methyl-l.4-dihydropyridine are obtained as a foam. ¹H-NMR (CDCl₃) : δ (TMS) 0.l-l.3 (6H, m): l.5-2.0 (4H, m; 2.l-3.0 (l0H, m); 3.20-4.20 (8H, m); 5.l0 (lH, s); 6.8 (lH, m) ; 6.9-7.4 (6H, m); 8.l (lH, s) Using the above described procedure the following compounds are prepared: 2-[(N-formyl-pyrrolidin-2-yl)methyl-thio]methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-metoxycarbonyl-4-(2-chlorophenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(2-trifluoromethylphenyl)-6-methyl-l,4-dihy dropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(3-cyanophenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)-3-ethoxycarbonyl-5-methoxycarbonyl-4-(3-methoxyphenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-phenyl-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(p-fluorophenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(2-trifluoromethylphenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3,5-di-ethoxycarbonyl-4-(3-pyridyl)-6-methyl-l,4-dihydropyridine; 2-(4-formylaminoethylthio)methyl-3,5-diethoxycarbonyl-4-(m-chlorophenyl)-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine, mp l09-lll°C 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-4-/ -benzo-(2,3-b)-l,4-dioxan-α-yl/-5-methoxycarbonyl-6-methyl-l,4-dihydropyridine; 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-(2-N-methyl-N-benzylamino)ethoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine. Example 7 A solution of 2-(2-aminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(2-chlorophenyl)-6-methyl-l,4-dihydropyridine (4.4 g) in THF (44 ml) is added to a stirred solution of N-formylimidazole, "in situ" prepared from formic acid (0.6 ml) and N,N-carbonyldiimidazole (2.5 g), at 0°C, in THF (25 ml). After 30 minutes the reaction mixture is poured into ice/water (2/l; 500 ml), extracted with AcOEt (3 × 50 ml), the combined extracts washed with a saturated solution of NaH₂PO₄ (3 × l0 ml), a saturated solution of NaHCO₃ (3 × l0 ml), and then with H₂O (3 × 50 ml), dried (Na₂SO₄) and concentrated under vacuum. 3.7 g of 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-carbomethoxy-4-(2-chlorophenyl)-6-methyl-l,4-dihydropyridine are obtained from Et₂O. Example 8 A solution of 2-[2-N-(2-cyanoethyl)aminoethyl]-thio]methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine(5.8 g) in N,N-dimethylformamide (l00 ml) and formic acid (l00 ml) is heated at l00°C for 6 hours, then poured in ice/water (500 ml) extracted with Et₂O(3 × 50 ml); the combined organic extracts are washed with a saturated solution of NaHCO₃ (3 × 30 ml), H₂O (3 × 20 ml), dried (Na₂SO₄) and evaporated to dryness. The residue is crystallized from EtOH to give 4.l5 g of 2-[2-N-(2-cyanoethyl)-N-formylaminoethylthio]methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine, mp ll0-ll2°C. Using the above described procedure, the following compounds are prepared. 2-(2-formylamino-2-ethoxycarbonylethylthio)methyl-3,5-diethoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-nitro-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine. Example 9 A solution of m-chloroperbenzoic acid (l.3 g, l equiv. mol) in l,2-dichloroethane (l5 ml) is added at -l0°C to a solution of 2-(2-formylaminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitro-phenyl)-6-methyl-l,4-dihydropyridine (3.5 g) in l,2-dichloroethane (30 ml). After 30 minutes the solution is filtered, washed with Na₂S₂O₃ (5% water solution, 3 × 5 ml), NaHCO₃ (saturated water solution, 3 × l0 ml), H₂O (3 × l0 ml), dried (Na₂SO₄) and evaporated in vacuum to give 3.4 g of 2-(2-formylaminoethylsulfinyl)methyl-3.ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine as a foam. lH-NMR (CDCl₃ δ (TMS) : l.00-l.2 (3H, t); 2.l (3H, t); 2.80-3.70 (4H, m); 3.90-4.30 (4H, m); 5.l0 (lH, m); 6.50 (lH, m); 7.l0-8.20 (6H, m). Using the above described procedure the following 2-(2-formylaminoethylsulfinyl)-6-methyl-l,4-dihydropyridines are prepared: -3-ethoxycarbonyl-5-cyano-4-(m-nitrophenyl); -3-ethoxycarbonyl-5-cyano-4-(m-chlorophenyl); -3,5-diethoxycarbonyl-4-(m-methylthiophenyl); -3,5-dimethoxycarbonyl-4-(m-trifluoromethylphenyl). Example l0 A solution of m-chloroperbenzoic acid (3.8 g, equiv. mol) in MeOH (30 ml) is added at l0°C to a solution of 2-(2-aminoethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine (5 g) in MeOH (l00 ml), the mixture is then warmed at l5-20°C and stirred for 30 minutes. The solvent is then evaporated at reduced pressure, the residue partitioned between CH₂Cl₂ (80 ml) and H₂O (30 ml), the organic phase is washed with sodium thiosulfate (5% water solution) and then worked-up as described in example 8 to give 4.8 g of 2-(2-aminoethyl-sulfonyl)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine as an amorphous solid. lH-NMR (CDCl₃) δ (TMS) l.0-l.2 (3H, t); 2.l (3H, t); 3.0-3.7 (4H, m), 4.l0-4.30 (4H, m); 5.l (lH, m); 6.8 (lH, m) 7.l0-8.20 (6H, m). Using the above described procedure the following compounds are prepared: 2-(2-formylaminoethylsulfonyl)-6-methyl-l,4-dihydropyridines: 3,5-diethoxycarbonyl-4-(m-methylthiophenyl) 3-ethoxycarbonyl-5-methoxyethoxycarbonyl-4-(m-chlorophenyl). Example ll Fractional crystallization of the (±)2-(2-ammoniummethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-dihydropyridine salt with (+) and (-)camphor-l0-sulfonic acids from EtOH produces 2 optical isomers: (+)free base [α] D = +l.8 (MeOH c = l0%) as (+)emifumarate salt m.p. l05-l07°C, [α] D = +3.6 (MeOH c = 9.7%) and (-)free base, [α] D = -l.7 (MeOH c = l9.8%), as emifumarate salt m.p. l06-l08°C, [α] D = 3.4 (MeOH c = 9.4%). Starting from the pure enantiomers, as free bases, using the procedures described in examples 7, 8, the enantiomeric 2-(2-formylaminomethylthio)methyl-3-ethoxycarbonyl-5-methoxycarbonyl-4-(m-nitrophenyl)-6-methyl-l,4-di hydropyridines are obtained as amorphous solids. (+)N-formylamino isomer: C₂₁H₃₅N₃O₇S. ½ H₂O [α] D = +l3.6, [α]₅₄₆ = +22.2 (MeOH, c = 2.l4) (-)N-formylamino isomer: C₂₁H₃₅N₃O₇S. ½ H₂O [α] D = -l3.7, [α]₅₄₆ = -22.3 (MeOH, c = 2.07)

Claims: Claim for the following Contracting States : ES, GR A process for the preparation of compounds of formula I wherein: R₁ represents: . a COCH₃, COC₆H₅, CN or NO₂ group; . a COORa group, wherein Ra represent hydrogen, C₁-C₆ straight or branched alkyl, optionally bearing one or more C₁-C₆ alkoxy groups and/or secondary amino groups of formula -NR₄R₅ wherein R₄ and R₅, that may be the same or different, represent C₁-C₆ alkyl, phenyl, benzyl or taken, together with the nitrogen atom, form a five or six membered ring optionally containing other atoms; R₂ is a phenyl ring unsubstituted or substituted with one or more C₁-C₆ alkyl, halogen, nitro, cyano, C₁-C₆ alkoxycarbonyl, C₁-C₆ alkylthio, C₁-C₆ alkylsulfinyl; pentafluorophenyl; α- or β-naphtyl; a five- or six-membered heterocyclic ring; α-benzo[2.3-b]-l.4-dioxan-α-yl; α-benzofuroxanyl; R₃ represents a COORa group wherein Ra is as above defined; P is selected from the group consisting of hydrogen, -(CH₂) p -W and C₁-C₈ linear or branched alkyl; N-P₁ is the residue of a primary or secondary amino group wherein P₁ is selected from the group consisting of hydrogen, C₁-C₆ lower linear or branched alkyl group and -(CH₂)p-W; P, taken together with P₁ and the nitrogen atom to which P₁ is linked, may form a pyrrolidine or a piperidine ring; W is selected from the group consisting of hydroxymethyl, formyloxymethyl, CO₂R wherein R is hydrogen or C₁-C₄ lower alkyl, CN, saturated or unsaturated heterocyclic ring, C₃-C₇ cycloalkyl ring, phenyl ring optionally substituted by one or more halogens or C₁-C₃-alkoxy groups; m is an integer from l to 3; n is zero or an integer from l to 2; p is zero or an integer from l to 3 comprising one of the following steps: a) forming a thioether bond by reaction of a compound of general formula (II) with a compound of formula III wherein: R₁, R₂, R₃, m, P, N-P₁ are as above defined; either Y or Y₁ represents a thiol or a masked thiol group such as thio C₂-C₁₂ alkanoyl ester or a thiouronium salt -S-(C=NR₆)NR₇R₈ (+) y₂ (-) the other being a known leaving group such as chlorine, bromine, iodine, trifluoromethane sulphonate, an alkyl- or an arylsulphonate; R₆, R₇, R₈, which are the same or different, are hydrogen or a C₁-C₄ alkyl group and y₂ (-) is a pharmaceutically acceptable anion; b) cyclizing a compound of formula IV with an alkylidene compound of formula V wherein: R₁, R₂, R₃, P, NP₁, m, are as above defined. c) cyclizing a compound of formula VI with an alkylidene compound of formula VII wherein R₂, R₃, P, NP₁ and m are as above defined and R₁, is prefarably CO₂Ra, CN, COCH₃, COC₆H₅. d) formylating a compound of formula VIII wherein R₁, R₂, R₃, P, P₁ and n are as above defined.

Inventors: FRIGERIO, MARCO, GANDOLFI, CARMELO A., SPINELLI, SILVANO, TOFANETTI, ODOARDO, TOGNELLA, SERGIO
Applicants: BOEHRINGER MANNHEIM ITALIA S.P.A.
Application year: 1987
Main classification: C07D211
IPC codes: C07D409, C07D401, C07D211, C07D405, C07D417, C07D413, C07D521