6H-Isoxazolo[3,4-d]pyrazolo[3,4-b]pyridines, a process for their preparation and their use as medicaments

Abstract: There are disclosed compounds having the formula wherein R₁ is hydrogen, loweralkyl, arylloweralkyl, aryl or heteroaryl; R₂ is hydrogen or loweralkyl; R₃ is loweralkyl, loweralkoxyloweralkyl, cycloalkyl, arylloweralkyl or heteroarylmethyl; and R₄ is hydrogen or loweralkyl; or a pharmaceutically acceptable acid addition salt thereof, which are useful as anxiolytic agents, methods for synthesizing them, and pharmaceutical compositions comprising an effective amount of such a compound.

Description: This invention relates to compounds having the formula wherein R₁ is hydrogen, C₁-C₆-alkyl, phenyl-C₁-C₆-alkyl or phenyl the phenyl nucleus being unsubstituted or substituted with 1, 2 or 3 substituents each of which being independently C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, trifluormethyl, chlorine or fluorine, with the proviso, that chlorine or fluorine are not at the ortho position or R₁ is a group of the formula where Y is O, S, NR₅ or -CH=N-, R₅ being hydrogen, C₁-C₆-alkyl or phenyl-C₁-C₆-alkyl; R₂ is hydrogen or C₁-C₆-alkyl; R₃ is C₁-C₆-alkyl, and R₄ is hydrogen or C₁-C₆-alkyl; or a pharmaceutically acceptable acid addition salt thereof, which are useful as anxiolytics, methods for synthesizing them and pharmaceutical compositions comprising an effective amount of such a compound. Throughout the specification and the appended claims, a given chemical formula or name shall encompass all stereo and optical isomers thereof where such isomers exist, as well as pharmaceutically acceptable acid addition salts thereof and solvates thereof such as for instance hydrates. The following definitions shall apply throughout the specification and the appended claims. Unless otherwise stated or indicated, the term C₁-C₆-alkyl denotes a straight or branched alkyl group having from 1 to 6 carbon atoms. Examples of said alkyl include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl and straight- and branched-chain pentyl and hexyl. Unless otherwise stated or indicated, the term C₁-C₆-alkoxy denotes a straight or branched alkoxy group having from 1 to 6 carbon atoms. Examples of said alkoxy include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy, t-butoxy and straight- and branched-chain pentoxy and hexoxy. Unless otherwise stated or indicated, the definition of the group having the formula where Y is O, S, NR₅ or CH=N, R₅ being hydrogen, loweralkyl or phenylloweralkyl, shall include all the positional isomers. Thus, for instance, the term shall include both 2-furyl and 3-furyl. Background art is: US-A-3 828 057 relating to 4-amino derivatives of pyrazolo[3,4-b]pyridine-5-ketones, which are central nervous system depressants and may be used as tranquilizers and FR-A-2 192 831 relating to derivatives of dipyrazolo[3,4-b; 3',4'-d]pyridines which, too, are central nervous system depressants and may be used as tranquilizers. Preferred embodiments of this invention are: a) A compound of formula (I), where R₁ is unsubstituted or substituted phenyl or C₁-C₆-alkyl, R₂ is hydrogen and R₄ is hydrogen. b) A compound as defined in a), where R₁ is C₁-C₆-alkyl, unsubstituted phenyl or phenyl which is substituted with C₁-C₄-alkyl, C₁-C₆-alkoxy, trifluoromethyl, fluorine or chlorine, with the proviso, that fluorine or chlorine are not in the ortho-position. c) A compound as defined in b), where R₃ is ethyl. The compounds of this invention are prepared by utilizing one or more of the steps described below. Throughout the description of the synthetic steps, the notations R₁, R₂, R₃, R₄, R₅ and Y shall have the respective meanings given above unless otherwise stated or indicated. Preferred synthetic steps used for synthesizing the compounds of this invention are schematically shown in the attached Reaction Scheme. For details of the reaction steps used for preparing compound III depicted in the Reaction Scheme, the reader is referred to H. Hoehn, Th. Denzel and W. Jansen, J. Heterocyclic Chem. 9 , 235 (1972), except that in this invention a low temperature (about -75° C to 40°C) Grignard reaction was used for the last step as shown in the Scheme, whereas Hoehn et al. used a cadmium-Grignard reagent at a higher temperature. STEP A Compound III where R₁ is not hydrogen or a group of the formula is reacted with hydroxylamine to obtain compound Ia This cyclization reaction is typically conducted by refluxing a mixture comprising compound III, hydroxylamine hydrochloride and a suitable solvent such as glacial acetic acid. It is preferable to add to the mixture an acid such as concentrated hydrochloric acid or p-toluenesulfonic acid. STEP B Compound II is reduced with a complex boronhydride or LiAlH₄ to obtain compound IV: Typically said reduction is conducted in a suitable medium such as an anhydrous ethereal solvent, e.g. tetrahydrofuran, diethyl ether or the like at a temperature of about 0-30° C. STEP C Compound IV is oxidized to compound V with manganese dioxide; Typically said oxidation is conducted in a suitable medium such as an aromatic hydrocarbon, e.g. benzene or toluene at a temperature of about 80-120° C. STEP D Compound V is reacted with heteroaryl lithium of formula VI where Z is O, S, NR₆ or CH=N, R₆ being C₁-C₆-alkyl, phenyl-C₁-C₆-alkyl, or benzenesulfonyl, to obtain compound VII: When Z is CH=N, the organolithium compound VI is prepared from n-butyl lithium and a bromo compound of the formula in a suitable solvent such as an anhydrous ethereal solvent, e.g. anhydrous diethylether at a temperature of between about -70° C and -50° C. When Z is O, S or NR₆, compound VI is prepared from n-butyl lithium and a compound of the formula in a suitable solvent such as anhydrous ether at a temperature of between about -20°C and +40° C. Thereafter the solution of compound VI prepared above is added to the solution a solution of compound V in a suitable solvent such as an anhydrous ethereal solvent, e.g. anhydrous tetrahydrofuran and the reaction mixture is stirred at a temperature of between about -20°C C and 10° C. When Z is benzenesulfonyl substituted nitrogen, 1-benzenesulfonylpyrrole which is used for preparing compound VI can readily be prepared from pyrrolyl anion and benzenesulfonyl chloride by routine procedure. STEP E Compound VII is oxidized with pyridinium chlorochromate to obtain compound VIII; This oxidation is typically conducted by adding pyridinium chlorochromate to a solution of compound VII in a suitable solvent such as a halogenated hydrocarbon, e.g. dichloromethane and stirring the mixture at a temperature of about 10-50° C. STEP F Compound VIII is reacted with hydroxylamine to obtain compound Ib; This step is conducted in substantially the same manner as STEP A. STEP G Although not indicated in the Reaction Scheme, when the group Z is benzenesulfonyl substituted nitrogen in formula Ib, the benzenesulfonyl group can be converted to hydrogen by hydrolizing the compound Ib. Said hydrolysis is conducted typically in the presence of an inorganic base such as NaOH or KOH and a suitable medium such as C₁-C₆-aliphatic alcohol at a temperature of about 80-120° C. As result of STEP F and STEP G, compound I where R₁ is a heteroaryl group of the formula , where Y is O, S, NR₅ or CH=N, R₅ being hydrogen, C₁-C₆-alkyl or phenyl-C₁-C₆-alkyl is obtained. STEP H Compound V is reacted with hydroxylamine to obtain compound Ic This step is conducted in substantially the same manner a STEP A above. STEP I Where the group R₁ in the target compound is a phenyl group substituted with one or more hydroxy groups, said compound is prepared by first synthesizing the corresponding C₁-C₆-alkoxy substituted compound according to the Reaction Scheme described above and then converting the alkoxy group or groups to hydroxy group or groups by cleavage reaction. Said cleavage reaction is typically conducted by refluxing a mixture comprising the alkoxy compound, boron tribromide and a suitable solvent such as a halogenated hydrocarbon 1,2-dichlorethane and thereafter pouring the reaction mixture into ice water. Compounds (I) of this invention are useful as anxiolytic agents. Anxiolytic activity was determined via direct interaction with benzodiazepin recognition sites in a membrane preparation from rat brain. Table I shows results of activity measurements for some of the compounds of this invention. Details of the procedure is described below: Procedure A. Reagents 1. ⊘.5 M Tris Buffer, pH 6.9: a. 78.1 g of Tris-HCl diluted to 1 liter b. 6⊘.6 g of Tris-base diluted to 1 liter c. Adjust PH of Tris-HCl to 6.9 at 25°C by adding Tris base. (⊘.5 M Tris buffer, pH 6.9) d. Make a 1:1⊘ dilution with distilled H₂O (⊘.⊘5 M Tris buffer, pH 6.9) 2. ⊘.32 M sucrose: 21.9 g of sucrose diluted to 2⊘⊘ ml. 3. [Methyl-³H]-Flunitrazepam (7⊘-9⊘ Ci/mmol) is procured from a commercial source. For IC 5⊘ determinations: ³H-Flunitrazepam is made up to a concentration of 8⊘ nM and 5⊘ micro-liter added to each tube (yields a final concentration of 2nM in the 2 ml assay volume). 4. Clonazepam HCl is obtained from a commercial source. A stock solution of ⊘.2 mM is made up in distilled H₂O. This is further diluted t⊘ ⊘.1 mM. Twenty micro-liter is added to 3 tubes to determine non specific binding (yields a final concentration of 1 x 1⊘ -6 M in the assay). 5. Test Compounds: For most assays, a 1 mM stock solution is made up in a suitable solvent and serially diluted, such that the final concentration in the assay ranges from 1⊘ -5 to 1⊘ -8 M. Seven concentrations are used for each assay and higher or lower concentrations may be used, depending on the potency of the drug. B. Tissue Preparation. Male Wistar rats are decapitated and the brain rapidly removed. The cerebral cortices are removed, weighed and homogenized with a homogenizer in 2⊘ volumes of ice-cold ⊘.32 M sucrose. This homogenate is centrifuged at 1⊘⊘⊘ g for 1⊘ minutes, the pellet is discarded and the supernatant is recentrifuged at 3⊘,⊘⊘⊘ g for 2⊘ minutes. The resulting membrane pellet is resuspended in 4⊘ volumes of ⊘.⊘5 M Tris buffer, pH 6.9. C. Assay 1 ml ⊘.⊘5 Tris buffer, pH 6.9 56⊘ micro-liter H₂O 7⊘ micro-liter ⊘.5 M Tris buffer, pH 6.9 5⊘ micro-liter ³H-Flunitrazepam 2⊘ micro-liter Vehicle (for total binding) or ⊘.1 mM Clonazepam (for nonspecific binding) or appropriate drug concentrations 3⊘⊘ micro-liter Tissue suspension The tubes containing ³H-flunitrazepam, buffer, drugs and H₂O are incubated at ⊘-4° C in an ice bath. A 3⊘⊘ micro-liter aliquot of the tissue suspension is added to the tubes at 1⊘-second intervals. The timer is started with the addition to the first tube. The tubes are then incubated for 2⊘ minutes at ⊘-4° C and the assay stopped by vacuum filtration of the samples through Whatman GF/B filters. This step is also done at 1⊘-second intervals. Each filter is immediately rinsed with three 5-ml washes of ice-cold Tris buffer, pH 6.9. Ths filters are counted in 1⊘ ml of Liquiscint (R) counting cocktail. Specific binding is defined as the difference between total binding and binding in the presence of 1 x 1⊘⁻⁶M clonazepam. Specific binding is approximately 97% of the total bound ligand. The percent inhibition at each drug concentration is the mean of triplicate determination. IC 5 calculations are performed using log-probit analysis. Effective quantities of the compounds of the invention may be administered to a patient by any of the various methods, for example, orally as in capsules or tablets, parenterally in the form of sterile solutions or suspensions, and in some cases intravenously in the form of sterile solutions. The free base final products, while effective themselves, may be formulated and administered in the form of their pharmaceutically acceptable acid addition salts for purposes of stablity, convenience of crystallization, increased solubility and the like. Acids useful for preparing the pharmaceutically acceptable acid addition salts of the invention include inorganic acids such as hydrochloric, hydrobromic, sulfuric, nitric, phosphoric and perchloric acids, as well as organic acids such as tartaric, citric, acetic, succinic, maleic, fumaric and oxalic acids. The active compounds of the present invention may be orally administered, for example, with an inert diluent or with an edible carrier, or they may be enclosed in gelatin capsules, or they may be compressed into tablets. For the purpose of oral therapeutic administration, the active compounds of the invention may be incorporated with excipients and used in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gum and the like. These preparations should contain at least ⊘.5% of active compound, but may be varied depending upon the particular form and may conveniently be between 4% to about 7⊘% of the weight of the unit. The amount of active compound in such compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that an oral dosage unit form contains between 1.⊘-3⊘⊘ milligrams of active compound. The tablets, pills, capsules, troches and the like may also contain the following ingredients: a binder such as micro-crystalline cellulose, gum tragacanth or gelatin; and excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel (R) , cornstarch and the like; a lubricant such as magnesium stearate or Sterotex (R) ; a glidant such as colloidal silicon dioxide; and a sweetening agent such as sucrose or saccharin may be added or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil. Other dosage unit forms may contain other various materials which modify the physical form of the dosage unit, for example, as coatings. Thus tablets or pills may be coated with sugar, shellac, or other enteric coating agents. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes, coloring and flavors. Materials used in preparing these various compositions should be pharmaceutically pure and non-toxic in the amounts used. For the purpose of parenteral therapeutic administration, the active compounds of the invention may be incorporated into a solution of suspension. These preparations should contain at least ⊘.1% of active compound, but may be varied between ⊘.5 and about 3⊘% of the weight thereof. The amount of active compound in such compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between ⊘.5 and 1⊘⊘ milligrams of active compound. The solutions or suspensions may also include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral multiple dose vials made of glass of plastic. Examples of the compounds of this invention include: 6-Ethyl-3-methyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-phenyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(3-fluorophenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(4-fluorophenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 3-(3-Chlorophenyl)-6-ethyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 3-(4-Chlorophenyl)-6-ethyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(2-tolyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(3-tolyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(4-tolyl)-6H-isoxazolo[3,4-d]pyraozlo[3,4-b]pyridine; 6-Ethyl-3-(4-trifluoromethylphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(2-methoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; 6-Ethyl-3-(3-methoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine; and 6-Ethyl-3-(4-methoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine. The following examples are presented in order to illustrate this invention. EXAMPLE 1 6-Ethyl-3-methyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 5-Acetyl-4-chloro-1-ethyl-1H-pyrazolo[3,4-b]pyridine (4.6⊘ g) was refluxed for 2 hours in 45⊘ ml of glacial acetic acid containing 15 g of hydroxylamine hydrochloride and 1.⊘ g of p-toluenesulfonic acid monohydrate. At the end of this time the reaction volume was reduced to ca. 2⊘⊘ ml under reduced pressure and then it was poured into 8⊘⊘ ml of water. The amorphous precipitate was filtered off and purified over 23⊘-4⊘⊘ mesh silica gel (5% ethyl acetate/dichloromethane). The appropriate fractions were combined and evaporated and the resultant crystalline product was washed with pentane to give 1.75 g, mp 181-182°C. ANALYSIS Calculated for C 1⊘ H 1⊘ N₄O: 59.39%C 4.98%H 27.71%N Found: 59.45%C 5.11%H 27.88%N EXAMPLE 2 6-Ethyl-3-phenyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 5-Benzoyl-4-chloro-1-ethyl-1H-pyrazolo[3,4-b]pyridine (5.⊘ g) as a slurry in 5⊘ ml of methanol was added to a solution of H₂NOH in 1⊘⊘ ml of methanol prepared by neutralizing 5.⊘ g of H₂NOH·HCl to a phenolphthalein endpoint. After stirring the mixture overnight the precipitated product was filtered off and dried, amounting to 1.65 g, mp 175-176°. The filtrate was evaporated and the residue taken up in dichloromethane. After a little insoluble material was filtered off, the residue was chromatographed over silica gel (5% ethyl acetate/dichloromethane) to give fractions containing both the end product and a new, more polar material. A few fractions that contained pure new material were evaporated; the new material was shown by ¹H-NMR, ¹³C-NMR, MS and IR to be the intermediate 5-benzoyl-4-hydroxyamino-1H-pyrazolo[3,4-b]pyridine. All the fractions containing the end product and this intermediate were evaporated the the residue was refluxed for 1 hour in 5⊘ ml of toluene containing 1⊘⊘ mg of p-toluenesulfonic acid, giving complete conversion to the end product. Workup with NaHCO₃/ether gave an additional ⊘.73 g of the end product, mp 175-176°. Material for analysis was recrystallized from methanol but the melting point was unchanged. ANALYSIS: Calculated for C₁₅H₁₂N₄O: 68.17%C 4.58%H 21.29%N Found: 68.⊘9%C 4.57%H 2⊘.97%N EXAMPLE 3 6-Ethyl-3-(3-fluorophenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(3-fluorobenzoyl)-1H-pyrazolo[3,4-b]pyridine (4.6⊘ g) and hydroxylamine hydrochloride (14.⊘ g) were refluxed in a solution of 25⊘ ml of acetic acid and 4 ml of concentrated hydrochloric acid. After 9⊘ minutes, the solvent was evaporated and the residue distributed between dichloromethane and an aqueous sodium bicarbonate solution. The organic phase was evaporated and the residue was chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane). Evaporation of the appropriate fractions gave 3.2 g of product, mp 182-184°. Recrystallization from ethyl acetate gave analytically pure product, mp 183-184°C. ANALYSIS: Calculated for C₁₅H₁₁FN₄O: 63.82%C 3.93%H 19.85%N Found: 63.75%C 4.⊘5%H 19.97%N EXAMPLE 4 6-Ethyl-3-(4-fluorophenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(4-fluorobenzoyl)-1H-pyrazolo[3,4-b]pyridine (4.5 g) and hydroxylamine hydrochloride (14.⊘ g) were refluxed in a solution of 25⊘ ml of acetic acid and 4 ml of concentrated hydrochloric acid. After two hours the solvent was evaporated and the residue distributed between dichloromethane and an aqueous sodium bicarbonate solution. The organic phase was evaporated and the residue chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane). Evaporation of the appropriate fractions gave 2.2 g of product after trituration with pentane, mp 222-223°C. ANALYSIS: Calculated for C₁₅H₁₁FN₄O: 63.82%C 3.93%H 19.85%N Found: 64.⊘7%C 4.⊘7%H 2⊘.13%N EXAMPLE 5 3-(3-Chlorophenyl)-6-ethyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-5-(3-chlorobenzoyl)-1-ethyl-1H-pyrazolo[3,4-b]pyridine (2.65 g) was stirred overnight at 9⊘° in 75 ml of glacial acetic acid containing 3.⊘ g of hydroxylamine hydrochloride. Sodium acetate (3.5 g) was then added, followed by an additional 1.⊘ g of hydroxylamine hydrochloride. The reaction mixture was then evaporated and the residue distributed between aqueous sodium bicarbonate and dichloromethane. The organic layer was evaporated and the residue was triturated with methanol and then chromatographed (23⊘-4⊘⊘ mesh silica gel, 1⊘ ethyl acetate/dichloromethane) to give 1.2⊘ g of product, mp 2⊘7-2⊘9°. It was combined with the products of several other runs and recrystallized from ethyl acetate to give analytically pure material, mp 2⊘9-21⊘°C. ANALYSIS: Calculated for C₁₅H₁₁ClN₄O: 6⊘.31%C 3.71%H 18.76%N Found: 6⊘.⊘6%C 3.77%H 18.68%N EXAMPLE 6 3-(4-Chlorophenyl)-6-ethyl-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-5-(4-chlorobenzoyl)-1-ethyl-1H-pyrazolo[3,4-b]pyridine (3.76 g) was suspended in 75 ml of glacial acetic acid, and to this was added 1.⊘5 g of hydroxylamine hydrochloride. The reaction mixture was stirred overnight at 8⊘° and the next morning yet another 1.⊘5 g of hydroxylamine hydrochloride was added. After stirring the mixture for additional two hours the solvent was evaporated and the residue washed first with sodium bicarbonate solution and then methanol. The resulting product was then chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane), giving 1.51 g of product, mp 248-25⊘°C. ANALYSIS: Calculated for C₁₅H₁₁ClN₄O: 6⊘.31%C 3.71%H 18.76%N Found: 6⊘.16%C 3.73%H 19.⊘3%N EXAMPLE 7 6-Ethyl-3-(2-tolyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(2-methylbenzoyl)-1H-pyrazolo[3,4-b]pyridine (5.4⊘ g) was refluxed for 2 hours in 3⊘⊘ ml of glacial acetic acid containing 1⊘ ml of concentrated hydrochloric acid and 1⊘ g of hydroxylamine hydrochloride. At the end of this time the reaction mixture was poured into water and extracted with dichloromethane. The organic phase was washed four times with water and once with saturated sodium bicarbonate solution. It was then evaporated and chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane) to give 2.21 g of product, mp 168-17⊘°. This product was recrystallized from ethyl acetate together with the product of another run to give analytically pure material, mp 17⊘-171°C. ANALYSIS: Calculated for C₁₆H₁₄N₄o; 69.⊘5%C 5.⊘7%H 2⊘.13%N Found: 68.94%C 5.15%H 2⊘.17%N EXAMPLE 8 6-Ethyl-3-(3-tolyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(3-methylbenzoyl)-1H-pyrazolo[3,4-b]pyridine (2.⊘ g) and hydroxylamine hydrochloride (6.⊘ g) were suspended in 2⊘⊘ ml of acetic acid, and to this was added ⊘.5 g of p-toluenesulfonic acid monohydrate. The reaction mixture was refluxed for 2 hours and then distributed between water and dichloromethane. The organic phase was washed four times with water and once with saturated sodium bicarbonate solution. It was then evaporated and chromatographed over 23⊘-4⊘⊘ mesh silica gel, eluting with 1⊘% ethyl acetate/dichloromethane. By combining the appropriate fractions, 1.4⊘ g of product was obtained, mp 178-179°. This product was combined with the product of another run and recrystallized from ethyl acetate to give product of mp 18⊘-181°C. ANALYSIS: Calculated for C₁₆H₁₄N₄O: 69.⊘4%C 5.⊘7%H 2⊘.13%N Found: 69.⊘3%C 5.16%H 2⊘.33%N EXAMPLE 9 6-Ethyl-3-(4-tolyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(4-methylbenzoyl)-1H-pyrazolo[3,4-b]pyridine (4.4⊘ g) and hydroxylamine hydrochloride (14.⊘ g) were mixed dry and then suspended in a solution prepared from 2⊘⊘ ml of glacial acetic acid and 5 ml of concentrated hydrochloric acid. The reaction mixture was then brought to reflux with mechanical stirring. After two hours, the reaction was allowed to cool and then the acetic acid was evaporated. The residue was distributed between dichloromethane and an aqueous sodium bicarbonate solution and then the organic phase was dried and evaporated. The crude product was chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane). The appropriate fractions were combined, evaporated, and then washed with pentane, giving 2.48 g of product, mp 221-222°C. ANALYSIS: Calculated for C₁₆H₁₄N₄O: 69.⊘5%C 5.⊘7%H 2⊘.13%N Found: 69.13%C 5.24%H 2⊘.41%N EXAMPLE 1⊘ 6-Ethyl-3-(4-trifluoromethylphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(4-trifluoromethylbenzoyl)-1H-pyrazolo[3,4-b]pyridine (3.1⊘ g) was refluxed for 2 hours in 3⊘⊘ ml of acetic acid containing 9 g of hydroxylamine hydrochloride and ⊘.6 g of p-toluenesulfonic acid monohydrate. At the end of this time the reaction mixture was poured into water and extracted with dichloromethane. The organic phase was washed well with water, dried, evaporated, and purified by chromatography over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane). In this way, 2.15 g of product was obtained, mp 243-245°. Recrystallization from ethyl acetate gave analytically pure product, mp 244-245°C. ANALYSIS: Calculated for C₁₆H₁₁F₃N₄O: 57.83%C 3.33%H 16.86%N Found: 58.26%C 3.32%H 16.75%N EXAMPLE 11 6-Ethyl-3-(2-methyoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine Five grams of 4-chloro-1-ethyl-5-(2-methoxybenzoyl)-1H-pyrazolo[3,4-b]pyridine was refluxed for 2 hours in 5⊘⊘ ml of acetic acid containing 15 g of hydroxylamine hydrochloride and 1 g of p -toluenesulfonic acid monohydrate. At the end of this time the reaction mixture was poured into water and extracted with dichloromethane. The organic phase was washed well with water, dried, evaporated and chromatographed over 23⊘-4⊘⊘ mesh silica gel. The appropriate fractions were combined and evaporated to give 2.95 g of product, mp 126-127°C. The melting point was unchanged after recrystallization from ethyl acetate/pentane. ANALYSIS: Calculated for C₁₆H₁₄N₄O₂: 65.29%C 4.79%H 19.⊘4%N Found: 65.63%C 4.99%H 19.39%N EXAMPLE 12 6-Ethyl-3-(3-methoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine Five grams of 4-chloro-1-ethyl-5-(3-methoxybenzoyl)-1H-pyrazolo[3,4-b]pyridine was refluxed for 2 hours in 5⊘⊘ ml of acetic acid containing 15 g of hydroxylamine hydrochloride and 1 g of p -toluenesulfonic acid monohydrate. At the end of this time the reaction mixture was poured into water and extracted with dichloromethane. The organic phase was washed well with water, dried, evaporated and chromatographed over 23⊘-4⊘⊘ mesh silica gel. The appropriate fractions were combined and evaporated to give 3.⊘5 g of product, mp 17⊘-171°. After recrystallization from water it had mp 169-171°C. ANALYSIS: Calculated for C₁₆H₁₄N₄O₂: 65.29%C 4.79%H 19.⊘4%N Found: 65.44%C 4.94%H 19.2⊘%N EXAMPLE 13 6-Ethyl-3-(4-methoxyphenyl)-6H-isoxazolo[3,4-d]pyrazolo[3,4-b]pyridine 4-Chloro-1-ethyl-5-(4-methoxybenzoyl)-1H-pyrazolo[3,4-b]pyridine (4.7⊘ g) was refluxed for 9⊘ minutes in 25⊘ ml of glacial acetic acid containing 15 ml of concentrated hydrochloric acid and 1⊘ g of hydroxylamine hydrochloride. At the end of this time the reaction mixture was poured into water and extracted with dichloromethane. The organic phase was washed four times with water and once with saturated sodium bicarbonate solution. It was then evaporated and chromatographed over 23⊘-4⊘⊘ mesh silica gel (1⊘% ethyl acetate/dichloromethane) to give 2.36 g (54%) of product, mp 215-216°. This product was recrystallized from ethyl acetate together with the product of another run to give analytically pure material, mp 215-216°C. ANALYSIS: Calculated for C₁₆H₁₄N₄O₂: 65.29%C 4.79%H 19.⊘4%N Found: 65.24%C 4.68%H 19.11%N EXAMPLE 14 4-Chloro-1-ethyl-1H-pyrazolo[3,4-b]pyridine-5-carboxaldehyde Wet MnO₂ (11⊘ g) was refluxed for 9⊘ minutes in 1 L of toluene with a Dean-Stark trap for the separation of water. This suspension was then cooled and 4-chloro-1-ethyl-1H-pyrazolo-[3,4-b]pyridine-5-methanol (17.5 g) was added in 5⊘ ml of toluene. The reaction mixture was then stirred for 6 hours and filtered with the aid of a filter pad. The toluene was evaporated and the residue added again to 1⊘⊘ g of wet MnO₂ freshly prepared in the above manner. After additional 4 hours the reaction was briefly warmed at 55° and then filtered again through a filter pad. The residue which remained after evaporation of the toluene was purified by preparative high performance liquid chromatography (1⊘% ethyl acetate/hexane, 25⊘ ml/min) to yield 11.7⊘ g of 4-chloro-1-ethyl-1H-pyrazolo[3,4-b]pyridine-5-carboxaldehyde, mp 89-9⊘°C. ANALYSIS : Calculated for C₉H₈ClN₃O: 51.56%C 3.85%H 2⊘.⊘%N Found: 51.18%C 3.79%H 19.84%N

Claims: Claims for the following Contracting States : BE, CH, DE, FR, GB, IT, LI, LU, NL, SE A compound having the formula where R₁ is hydrogen, C₁-C₆-alkyl, phenyl-C₁-C₆-alkyl, or phenyl the phenyl nucleus being unsubstituted or substituted with 1, 2 or 3 substituents each of which being independently C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, trifluoromethyl, chlorine or fluorine, with the proviso that chlorine or fluorine are not at the ortho position or R₁ is a group having the formula where Y is O, S, NR₅ or CH=N, R₅ being hydrogen, C₁-C₆-alkyl or phenyl-C₁-C₆-alkyl; R₂ is hydrogen or C₁-C₆-alkyl, R₃ is C₁-C₆-alkyl; and R₄ is hydrogen or C₁-C₆-alkyl and pharmaceutical acceptable acid addition salts thereof. A compound as defined in claim 1, where R₁ is unsubstituted or substituted phenyl or C₁-C₆-alkyl, R₂ is hydrogen and R₄ is hydrogen. A compound as defined in claim 2, where R₁ is C₁-C₆-alkyl, unsubstituted phenyl or phenyl which is substituted with C₁-C₆-alkyl, C₁-C₆-alkoxy, trifluoromethyl, fluorine or chlorine, with the proviso, that fluorine or chlorine are not in the ortho-position. A compound as defined in claim 3, where R₃ is ethyl. The compound as defined in claim 1 which is 6-ethyl-3-phenyl-6 H-isoxazolo-[3,4-d]-pyrazolo-[3,4-b] pyridine, or a pharmaceutically acceptable acid addition salt thereof. The compound as defined in claim 1 which is 6-ethyl-3-(4-chlorophenyl)-6 H-isoxazolo [3,4-d]pyrazolo[3,4-b]-pyridine, or a pharmaceutically acceptable acid addition salt thereof. The compound as defined in claim 1 which is 6-ethyl-3-(4-fluorophenyl)-6 H-isoxazolo [3,4-d]pyrazolo[3,4-b]-pyridine, or a pharmaceutically acceptable acid addition salt thereof. The compound as defined in claim 1 which is 6-ethyl-3-(4-tolyl)-6 H - isoxazolo-[3,4-d]pyrazolo [3,4-b]pyridine, or a pharmaceutically acceptable acid addition salt thereof. A pharmaceutical composition which comprises as active ingredient a compound as defined in claim 1 in association with a pharmaceutically acceptable carrier. Use of a compound as defined in claim 1 for the preparation of a medicament having anxiolytic activity. A process for the preparation of a compound of formula I as defined in claim 1 which comprises a) reacting a compound of formula IIIa wherein R₁, R₂, R₃ and R₄ are as defined above, with hydroxylamine, b) optionally converting a compound of formula I wherein R₁ is phenyl substituted with one or more C₁-₆-alkoxy groups by means of a cleavage reaction to a compound of formula I wherein R₁ is phenyl substituted with one or more hydroxy groups, and c) optionally preparing in a conventional manner a pharmaceutically acceptable acid addition salt thereof. A process as defined in claim 11 where R₁ is C₁-C₆-alkyl or unsubstituted or substituted phenyl, R₂ is hydrogen and R₄ is hydrogen. A process as defined in claim 12 wherein R₁ is C₁-C₆-alkyl, unsubstituted phenyl or phenyl substituted with C₁-C₆-alkyl, C₁-C₆-alkoxy, trifluormethyl, fluorine or chlorine, with the proviso that fluorine or chlorine are not in the ortho-position. A process as defined in claim 13 wherein R₃ is ethyl. Claims for the following Contracting States : AT, ES, GR A process for the preparation of a compound of formula I where R₁ is hydrogen, C₁-C₆-alkyl, phenyl-C₁-C₆-alkyl, or phenyl the phenyl nucleus being unsubstituted or substituted with 1, 2 or 3 substituents each of which being independently C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, trifluoromethyl, chlorine or fluorine, with the proviso that chlorine or fluorine are not at the ortho position, or R₁ is a group having the formula where Y is O, S, NR₅ or CH=N, R₅ being hydrogen, C₁-C₆-alkyl or phenyl-C₁-C₆-alkyl; R₂ is hydrogen or C₁-C₆-alkyl, R₃ is C₁-C₆-alkyl; and R₄ is hydrogen or C₁-C₆-alkyl and pharmaceutical acceptable acid addition salts thereof, which comprises which comprises a) reacting a compound of formula IIIa wherein R₁, R₂, R₃ and R₄ are as defined above, with hydroxylamine, b) optionally converting a compound of formula I wherein R₁ is phenyl substituted with one or more c₁-C₆-alkoxy groups by means of a cleavage reaction to a compound of formula I wherein R₁ is phenyl substituted with one or more hydroxy groups, and c) optionally preparing in a conventional manner a pharmaceutically acceptable acid addition salt thereof. A process as defined in claim 1 where R₁ is C₁-C₆-alkyl or unsubstituted or substituted phenyl, R₂ is hydrogen and R₄ is hydrogen. A process as defined in claim 2 wherein R₁ is C₁-C₆-alkyl, unsubstituted phenyl or phenyl substituted with C₁-C₆-alkyl, C₁-C₆-alkoxy, trifluormethyl, fluorine or chlorine, with the proviso that fluorine or chlorine are not in the ortho-position. A process as defined in claim 3 wherein R₃ is ethyl.

Inventors: SHUTSKE, GREGORY M.
Applicants: HOECHST-ROUSSEL PHARMACEUTICALS INCORPORATED
Application year: 1986
Main classification: C07D498
IPC codes: A61K31, C07D498, A61P25