PHOSPHODIESTERASE INHIBITORS
Field of the Inyention
The present inyention relates to phosphodiesterase (PDE) type IV selective inhibitors. Compounds disclosed herein can be useful in the treatment of CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease (COPD), psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome (ARDS), eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases especially in humans. Processes for the preparation of disclosed compounds, pharmaceutical compositions containing the disclosed compounds, and their use as PDE type IV selective inhibitors, are provided.
Background of the Inyention
It is known that cyclic adenosine-3', 5'-monophosphate (cAMP) exhibits an important role of acting as an intracellular secondary messenger (E W. Sutherland, and TW Roll. Pharmacol Rev, 1960,12, 265) Its intracellular hydrolysis to adenosine 5'-monophosphate (AMP) causes a number of inflammatory conditions which are not limited to psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome (ARDS), eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis PDE4 inhibitors are designed to inhibit the activity of PDE4, the enzyme which breaks down neuronal cAMP Studies have shown that administering PDE4 inhibitors can have a restorative effect on memory loss in animal models, including those of Alzheimer's disease (Expert Opin. Ther Targets (2005) 9(6) 1283-1305, Drug Discovery today, vol 10, number 22, November 2005). The most important role in the control of cAMP (as well as of cGMP) level is played by cyclic nucleotide phosphodiesterases (PDE) which represent a biochemically and functionally highly variable super family of enzymes, eleven distinct families with more than 25 gene products are currently recognized Although PDE I, PDE II, PDE III, PDE IV, and PDE VII all use cAMP as a substrate, only PDE IV and PDE VII are highly selective for hydrolysis of cAMP Inhibitors of PDE, particularly the PDE IV inhibitors, such as rolipram or Ro-1724 are therefore known as cAMP-enhancers. Immune cells contain type IV and type III PDE, the PDE IV type being prevalent in human mononuclear cells. Thus the inhibition of phosphodiesterase type IV has been a target for modulation and, accordingly, for therapeutic intervention in a range of disease processes
The initial observation that xanthine derivatives, theophylline and caffeine inhibit the hydrolysis of c AMP led to the discovery of the required hydrolytic activity in the cyclic nucleotide phosphodiesterase (PDE) enzymes Distinct classes of PDE's have been recognized (J A Beavo and D H. Reifsnyder, TIPS, 1990,11,150), and their selective inhibition has led to improved drug therapy (C D Nicholus, R A Chalhss and M. Shahid, TIPS, 1991, 12, 19) Thus it was recognized
that inhibition of PDEIV could lead to inhibition of inflammatory mediator release (M W. Verghese et al, J Mol. Cell. Cardiol. 1989,12 (Suppl II), S 61) and airway smooth muscle relaxation
U S Patent No 5,686,434 discloses 3-aryl-2-isoxazohnes as anti-inflammatory agents U.S. Patent Nos. 6,114,367 and 5, 869,511 disclose isoxazohne compounds as inhibitors of TNF release. WO 95/14681 discloses a series of isoxazohne compounds as anti-inflammatory agents WO 02/100332 discloses isoxazohne compounds having macrophage inhibitory factor (MIF) antagonist activity.
Summary of the Inyention
The present inyention provides phosphodiesterase inhibitors, which can be used for the treatment of CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease (COPD), psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome (ARDS), eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases, and the processes for the synthesis of these compounds.
Pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides of these compounds having the same type of activity are also provided
Pharmaceutical compositions containing the compounds, which may also contain pharmaceutically acceptable carriers or diluents, can be used for the treatment of CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease (COPD), psonasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome, eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases
Other aspects will be set forth in the accompanying description which follows and in part will be apparent from the description or may be learnt by the practice of the inyention
In accordance with one aspect, there are provided compounds having the structure of Formula I
(Formula Removed)

their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides, wherein
R1, R2 and R3 can be independently selected from hydrogen or alkyl,
X1 and X2 can be independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl, or (heterocyclyl)alkyl;
Y can represent an oxygen atom, a sulphur atom or NR (wherein R can be selected from hydrogen, alkyl, alkenyl, alkynyl, un(saturated) cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl, or (heterocyclyl)alkyl),
Y1 and Y2 can be independently selected from hydrogen, alkyl, nitro, cyano, halogen, OR (wherein R can be the same as defined earlier), SR (wherein R can be the same as defined earlier), NHR (wherein R can be the same as defined earlier), COOR' or COR' (wherein R' can be hydrogen, alkyl, alkenyl, alkynyl, (un)saturated cycloalkyl, aryl, aralkyl, heterocyclyl, (heterocyclyl)alkyl, or (heteroaryl)alkyl);
Further, Y1 and X2, X1 and Y2, X1 and X2 may together form a cyclic ring fused with the ring, containing 3-5 carbon atoms within the nng and having 1-3 heteroatoms selected from N, O or S
The following definitions apply to terms as used herein-
The term "alkyl," unless otherwise specified, refers to a monoradical branched or unbranched saturated hydrocarbon chain having from 1 to 20 carbon atoms. This term can be exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n-decyl, tetradecyl, and the like. Alkyl groups may be substituted further with one or more substituents selected from alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, carboxyalkyl, aryl, heterocyclyl, heteroaryl, arylthio, thiol, alkylthio, aryloxy, mtro, aminosulfonyl, aminocarbonylamino, -NHC(=O)Rf, -NRfRq, -C(=O)NH2, -COOR" (wherein R" is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl), -C(=O)NRfRq, -NHC(=O)NRfRq,, -C(=O)heteroaryl, C(=O)heterocyclyl, -O-C(=O)NRfRq {wherein Rf and Rq are independently selected from alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, aralkyl, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, (heteroaryl)alkyl}, nitro, or -SO2R6 (wherein R6 is alkyl, alkenyl, alkynyl, cycloalkyl, aralkyl, aryl, heterocyclyl, heteroaryl, (heteroaryl)alkyl or (heterocyclyl)alkyl). Unless otherwise constrained by the definition, alkyl substituents may be further substituted by 1-3 substituents selected from alkyl, carboxy, -NRfRq, -C(=O)NRfRq, -OC(=O) NRfRq , -NHC(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), hydroxy, alkoxy, halogen, CF3, cyano, and -SO2R6, (wherein R6 is the same as defined earlier), or an alkyl group also may be interrupted by 1-5 atoms of groups independently selected from oxygen, sulfur or -NRa- {wherein Ra is selected from hydrogen, alkyl,
cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, acyl, aralkyl, -C(=O)ORf (wherein Rf is the same as defined earlier), SO2R6 (where R6 is as defined earlier), or -C(=O)NRfRq (wherein Rf and Rq are as defined earlier)} Unless otherwise constrained by the definition, all substituents may be substituted further by 1-3 substituents selected from alkyl, carboxy, -NRfRq, -C(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier) hydroxy, alkoxy, halogen, CF3, cyano, and -SO2R6 (where R6 is same as defined earlier), or an alkyl group as defined above that has both substituents as defined above and is also interrupted by 1-5 atoms or groups as defined above
The term "alkenyl," unless otherwise specified, refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group having from 2 to 20 carbon atoms with cis, trans, or geminal geometry. In the event that alkenyl is attached to a heteroatom, the double bond cannot be alpha to the heteroatom Alkenyl groups may be substituted further with one or more substituents selected from alkyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, -NHC (=O)Rf, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, (heteroaryl)alkyl, aminosulfonyl, ammocarbonylamino, alkoxyamino, mtro, or SO2R6 (wherein R6 is same as defined earlier) Unless otherwise constrained by the definition, alkenyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, carboxy, hydroxy, alkoxy, halogen, -CF3, cyano, -NRfRq, -C(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier) and -SO2R6 (where R6 is same as defined earlier).
The term "alkynyl," unless otherwise specified, refers to a monoradical of an unsaturated hydrocarbon, having from 2 to 20 carbon atoms In the event that alkynyl is attached to a heteroatom, the triple bond cannot be alpha to the heteroatom. Alkynyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl, ammocarbonylamino, mtro, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, (heteroaryl)alkyl, -NHC(=O)Rf, -NRfRq, -NHC(=O)NRfRq , -C(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), or -SO2R6 (wherein R6 is as defined earlier). Unless otherwise constrained by the definition, alkynyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, carboxy, carboxyalkyl, hydroxy, alkoxy, halogen, CF3, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq, -C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), cyano, or -SO2R6 (where R6 is same as defined earlier)
The term "cycloalkyl," unless otherwise specified, refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings, which may optionally contain one or more olefinic bonds, unless otherwise constrained by the definition. Such cycloalkyl groups can include, for example, single ring structures, including cyclopropyl, cyclobutyl, cyclooctyl, cyclopentenyl, and the like, or multiple ring structures, including adamantanyl, and bicyclo [2.2 1] heptane, or cyclic alkyl groups to which is fused an aryl group, for example, indane, and the like. Spiro and fused ring structures can also be included Cycloalkyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, carboxyalkyl, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl, aminocarbonylamino, -NRfRq, -NHC(=O)NRfRq, -NHC(=O)Rf, -C(=O)NRfRq, -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), nitro, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, (heteroaryl)alkyl, or -SO2R6 (wherein R6 is same as defined earlier) Unless otherwise constrained by the definition, cycloalkyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, carboxy, hydroxy, alkoxy, halogen, CF3, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq , -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), cyano or -SO2R6 (where R$ is same as defined earlier)
The term "alkoxy" denotes the group O-alkyl, wherein alkyl is the same as defined above
The term "aryl," unless otherwise specified, refers to carbocyclic aromatic groups, for example, phenyl, biphenyl or napthyl ring and the like, optionally substituted with 1 to 3 substituents selected from halogen (e g, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, acyl, aryloxy, CF3, cyano, nitro, COORe (wherein Re is hydrogen, alkyl, alkenyl, cycloalkyl, aralkyl, (heterocyclyl)alkyl, (heteroaryl)alkyl), NHC(=O)Rf, -NRfRq, -C(=O)NRfRq, -NHC(=O)NRfRq , -O-C(=O)NRfRq (wherein Rf and Rq are the same as defined earlier), -SO2R6 (wherein R6 is same as defined earlier), carboxy, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, (heteroaryl)alkyl or amino carbonyl amino The aryl group optionally may be fused with a cycloalkyl group, wherein the cycloalkyl group may optionally contain heteroatoms selected from O, N or S
The term "aralkyl," unless otherwise specified, refers to alkyl-aryl linked through an alkyl portion (wherein alkyl is as defined above) and the alkyl portion contains 1-6 carbon atoms and aryl is as defined below Examples of aralkyl groups include benzyl, ethylphenyl and the like.
The term "aralkenyl," unless otherwise specified, refers to alkenyl-aryl linked through alkenyl (wherein alkenyl is as defined above) portion and the alkenyl portion contains 1 to 6 carbon atoms and aryl is as defined below
The term "aryloxy" denotes the group O-aryl, wherein aryl is as defined above
The term "carboxy," as defined herein, refers to -C(=O)OH
The term "heteroaryl," unless otherwise specified, refers to an aromatic ring structure containing 5 or 6 ring atoms, or a bicyclic aromatic group having from 8 to 10 ring atoms, with one or more heteroatom(s) independently selected from N, O or S optionally substituted with 1 to 4 substituent(s) selected from halogen (e g, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, carboxy, aryl, alkoxy, aralkyl, cyano, nitro, heterocyclyl, heteroaryl, -NRfRq, CH=NOH, -(CH2)wC(=O)Rg {wherein w is an integer from 0-4 and Rg is hydrogen, hydroxy, ORf, NRfRq, -NHORz or -NHOH}, -C(=O)NRfRq and -NHC(=O)NRfRq, -SO2R6, -O-C(=O)NRfRq, -O-C(=O)Rf, -O-C(=O)ORf (wherein R6, Rf and Rq are as defined earlier, and Rz is alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl). Unless otherwise constrained by the definition, the substituents are attached to a ring atom, / e, carbon or heteroatom in the ring Examples of heteroaryl groups include oxazolyl, lmidazolyl, pyrrolyl, 1,2,3-tnazolyl, 1,2,4-tnazolyl, tetrazolyl, thiazolyl, oxadiazolyl, benzoimidazolyl, thiadiazolyl, pyridinyl, pyndazinyl, pynmidinyl, pyrazinyl, thienyl, isoxazolyl, tnazinyl, furanyl, benzofuranyl, indolyl, benzothiazolyl, or benzoxazolyl, and the like
The term 'heterocyclyl," unless otherwise specified, refers to a non-aromatic monocyclic or bicyclic cycloalkyl group having 3 to 10 atoms wherein 1 to 4 carbon atoms in a ring are replaced by heteroatoms selected from O, S or N, and their oxidized forms, and optionally are benzofused or fused heteroaryl having 5-6 ring members and/or optionally are substituted, wherein the substituents are selected from halogen (eg, F, CI, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, aryl, alkoxy, alkaryl, cyano, nitro, oxo, carboxy, heterocyclyl, heteroaryl, -O-C(=O)Rf, -O-C(=O)ORf, -C(=O)NRfRq, SO2R6, -O-C(=O)NRfRq, -NHC(=O)NRfRq, -NRfRq (wherein R6, Rf and Rq are as defined earlier) or guanidine Heterocyclyl can optionally include rings having one or more double bonds Unless otherwise constrained by the definition, the substituents are attached to the ring atom, / e, carbon or heteroatom in the ring. Also, unless otherwise constrained by the definition, the heterocyclyl ring optionally may contain one or more olefinic bond(s). Examples of heterocyclyl groups include oxazohdmyl, tetrahydrofuranyl, dihydrofuranyl, dihydropyndinyl, dihydroisoxazolyl, dihydrobenzofuryl, azabicyclohexyl, dihydroindolyl, isoindole 1,3-dione, pipendinyl, morphohnyl or piperazinyl
"(Heteroaryl)alkyl" refers to alkyl-heteroaryl group linked through alkyl portion, wherein the alkyl and heteroaryl are as defined earlier
"(Heterocyclyl)alkyl" refers to alkyl-heterocyclyl group linked through alkyl portion, wherein the alkyl and heterocyclyl are as defined earlier
"Acyl" refers to -C(=O)R', wherein R' is selected from hydrogen, alkyl, alkenyl, alkynyl, (un)saturated cycloalkyl, aryl, aralkyl, heterocyclyl, (heterocyclyl)alkyl, or (heteroaryl)alkyl.
"Alkylcarbonyl" refers to -C(=O)R", wherein R" is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl
"Alkylcarboxy" refers to -O-C(=O)R", wherein R" is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl
"Amine," unless otherwise specified, refers to -NH2 "Substituted amine," unless otherwise specified, refers to -N (Rk)2, wherein each Rk independently is selected from hydrogen {provided that both Rk groups are not hydrogen (defined as "amino")}, alkyl, alkenyl, alkynyl, aralkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, (heterocyclyl)alkyl, (heteroaryl)alkyl, acyl, SO2R6 (wherein Re is as defined above), -C(=O)NRfRq, NHC(=O)NRfRq, or -NHC(=O)0Rf (wherein Rf and Rq are as defined earlier).
"Thiocarbonyl" refers to -C(=S)H "Substituted thiocarbonyl" refers to-C(=S)R", wherein R" is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl, amine or substituted amine
Unless otherwise constrained by the definition, all substituents optionally may be substituted further by 1-3 substituents selected from alkyl, aralkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, carboxy, carboxyalkyl, hydroxy, alkoxy, halogen, CF3, cyano, -C(=O)NRfRq, -0(C=0)NRfRq (wherein Rf and Rq are the same as defined earlier) and -SO2R6 (where R6 is the same as defined earlier)
The term "leaving group" refers to groups that exhibit or potentially exhibit the properties of being labile under the synthetic conditions and also, of being readily separated from synthetic products under defined conditions Examples of leaving groups include, but are not limited to, halogen (e g, F, CI, Br, I), triflates, tosylate, mesylates, alkoxy, thioalkoxy, or hydroxy radicals and the like
The term "protecting groups" refers to moieties that prevent chemical reaction at a location of a molecule intended to be left unaffected during chemical modification of such molecule Unless otherwise specified, protecting groups may be used on groups, such as hydroxy, amino, or carboxy Examples of protecting groups are found in T W Greene and P.G M. Wuts, "Protective Groups in Organic Synthesis", 2nd Ed, John Wiley and Sons, New York, NY., which is incorporated herein by reference The species of the carboxyhc protecting groups, amino protecting groups or hydroxy protecting groups employed are not critical, as long as the denyatised moieties/moiety is/are stable to conditions of subsequent reactions and can be removed without disrupting the remainder of the molecule Certain "protecting groups" may be formed in situ under the reaction conditions and may be removed when the conditions under which they are formed are modified Example of such protection is the lithiation of hydroxyl groups under hthiation
conditions
The term "pharmaceutically acceptable salts" refers to derivatives of compounds that can be modified by forming their corresponding acid or base salts Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acids salts of basic residues (such as amines), or alkali or organic salts of acidic residues (such as carboxylic acids), and the like.
The compounds provided herein can be used for treating CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease, psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome, eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases
In accordance with yet another aspect, there are provided processes for the preparation of the compounds as described herein
Detailed Description of the Inyention
The compounds described herein may be prepared by techniques well known in the art and familiar to the average synthetic organic chemist In addition, the compounds of present inyention may be prepared by the following reaction sequjgBfieg as depicted in schemes I, II, III IV and V
(Scheme Removed)
The compounds of Formula IX can be prepared by following Scheme I Accordingly, reacting a compound of Formula II with a compound of Formula XjZ (wherein Z is halogen) can give a compound of Formula III [wherein X1 (except hydrogen), Y1 and Y2 are the same as defined earlier], which on reaction with a compound of Formula X2Z [wherein Z is halogen] can give a compound of Formula IV [wherein X2 (except hydrogen) is same as defined earlier], which on reaction with hydroxylamme hydrochloride can give a compound of Formula V, which on treatment with a compound of Formula VI can give a compound of Formula VII [wherein R\ and R2 are the same as defined earlier and Rr represents COOH, COOCH3], which (when Rr is
COOCH3) on reaction with hydrazine hydrate can give a compound of Formula VIII, which can finally be reacted with a compound of Formula HC(OR.3)3 to give a compound of Formula IX [wherein R3 is the same as defined earlier].
The reaction of a compound of Formula II with a compound of Formula X\Z to give a compound of Formula III can be carried out in the presence of one or more of phase transfer catalysts, for example, benzyltnbutyl ammonium chloride, benzyltriethylammonium chloride, benzyltnethylammonium iodide or mixtures thereof.
The reaction of a compound of Formula II with a compound of Formula X\Z can be carried out in the presence of one or more of inorganic bases, for example, alkali metal hydroxides, for example, sodium hydroxide, potassium hydroxide, lithium hydroxide, alkali metal carbonates, for example, potassium carbonate, cesium carbonate or mixtures thereof
The reaction of a compound of Formula II with a compound of Formula X\Z can be carried out in one or more of solvents, for example, tetrahydrofuran, dimethylformamide, dimethylsulphoxide, acetomtnle, dimethylacetamide or mixtures thereof
The reaction of a compound of Formula III with a compound of Formula X2Z can be earned out in the presence of one or more of inorganic bases, for example, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, cesium carbonate or mixtures thereof
The reaction of a compound of Formula III with a compound of Formula X2Z to give a compound of Formula IV can be carried out in one or more of solvents, for example, tetrahydrofuran, dimethylformamide, dimethylsulphoxide, acetomtrile, acetone, dimethylacetamide or mixtures thereof
The reaction of a compound of Formula IV with hydroxylamine hydrochloride to give a compound of Formula V can be carried out in the presence of sodium acetate, potassium acetate, tnethylamine or pyridine in one or more of solvents, for example, methanol, ethanol, propanol, n-butanol or mixtures thereof
The reaction of a compound of Formula V with a compound of Formula VI to give a compound of Formula VII can be earned out in the presence of sodium hypochlorite in one or more of solvents, for example, tetrahydrofuran, dimethylformamide, dimethylsulphoxide, acetonitnle, chloroform, dichloromethane or mixtures thereof
The reaction of a compound of Formula VII with hydrazine hydrate to give a compound of Formula VIII can be carried out at a temperature ranging, for example, from 120 to 140°C
The reaction of a compound of Formula VIII with a compound of Formula HC(OR.3)3 to give a compound of Formula IX can be earned out at a temperature ranging, for example, from 60 to 160°C
(Scheme Removed)
The compounds of Formula IX can also be prepared by following Scheme II Accordingly, a compound of Formula X on debenzylation can give a compound of Formula XI [wherein X1, Y1, Y2, R1, R2 and R3 are the same as defined earlier], which, finally on reaction with X2Z [wherein Z is halogen] can give a compound of Formula IX [wherein X2 (except hydrogen and benzyl) is same as defined earlier].
The debenzylation of a compound of Formula X to give a compound of Formula XI can be carried out by catalytic transfer hydrogenation in the presence of one or more of palladium catalysts or ammonium formate or in the presence of boron tribromide in one or more of solvents, for example, methanol, ethanol, propanol, n-butanol, toluene or mixtures thereof
The reaction of a compound of Formula XI with a compound of Formula X2Z to give a compound of Formula IX can be carried out in the presence of one or more of inorganic bases, for example, sodium carbonate, sodium bicarbonate, potassium carbonate, cesium carbonate, potassium bicarbonate or mixtures thereof
The reaction of a compound of Formula XI with a compound of Formula X2Z can be earned out in one or more of solvents, for example, tetrahydrofuran, dimethylformamide, dimethylsulphoxide, acetonitnle, acetone, dimethylacetamide or mixtures thereof
(Scheme Removed)
The compounds of Formula XIII can be prepared by following Scheme III. Accordingly, a compound of Formula XII can be amidated to give a compound of Formula XIII [wherein Xj, Y1, Y2, R1, R2 and R3 are the same as defined earlier].
The amidation of a compound of Formula XII to give a compound of Formula XIII can be carried out in the presence of methanolic ammonia or an alkylamine
(Scheme Removed)
The compounds of Formula XVI and Formula XVII can be prepared by following Scheme IV Accordingly, a compound of Formula XIV can be reacted with a compound of Formula XV to give a compound of Formula XVI [wherein X1, X2, Y\, Y2 and Ri are the same as defined earlier] and a compound of Formula XVII [wherein X1, X2, Y1, Y2 and Rj are the same as defined earlier].
The reaction of a compound of Formula XIV with a compound of Formula XV to give a compound of Formula XVI and a compound of Formula XVII can be carried out in the presence of one or more of halogenatmg agents, for example, thionyl chloride, oxalyl chloride, sulfuryl chloride, phosphorus oxychloride, phosphorus trichloride, phosphorus pentachlonde, phosphorus tnbromide or mixtures thereof
The reaction of a compound of Formula XIV with a compound of Formula XV can be carried out in one or more of solvents, for example, benzene, toluene, dichloromethane, chloroform or mixtures thereof
(Scheme Removed)
The compounds of Formula XXIII can be prepared by following Scheme V Accordingly, a compound of Formula XVIII [wherein configuration at stereogenic carbons marked * can be (R) or (S)] on reaction with hydrazine hydrate can give a compound of Formula XIX, which on reaction with methanol can give a compound of Formula XX, which on reaction with Freon gas can give a compound of Formula XXI, which on reaction with hydrazine hydrate can give a compound of
Formula XXII, which can, finally, be reacted with a compound of Formula HC(OR3)3 to give a compound of Formula XXIII [wherein X2, Y1, Y2, Ri and R3 are the same as defined earlier and configuration at stereogenic carbon marked * can be (R) or (S)]
The reaction of a compound of Formula XVIII with hydrazine hydrate to give a compound of Formula XIX can be carried out in the presence of one or more of inorganic bases, for example, potassium hydroxide, sodium hydroxide, lithium hydroxide, cesium hydroxide or mixtures thereof.
The reaction of a compound of Formula XVIII with hydrazine hydrate can be carried out in one or more of solvents, for example, methanol, ethanol, propanol, isopropanol, ethylene glycol or mixtures thereof.
The reaction of a compound of Formula XIX with methanol to give a compound of Formula
XX can be carried out in the presence of one or more of mineral acids, for example, sulphuric acid,
hydrochloric acid or mixtures thereof
The reaction of a compound of Formula XX with Freon gas to give a compound of Formula
XXI can be carried out in the presence of one or more of phase transfer catalysts, for example,
benzyltributylammonium chloride, benzyltnethylammonium chloride, benzyltnethylammonium
iodide or mixtures thereof
The reaction of a compound of Formula XX with Freon gas can be carried out in the presence of one or more of inorganic bases, for example, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, cesium carbonate or mixtures thereof.
The reaction of a compound of Formula XX with Freon gas can be carried out in one or more of solvents, for example, tetrahydrofuran, dimethylformamide, dimethylsulphoxide, acetonitnle, dimethylacetamide or mixtures thereof
The reaction of a compound of Formula XXI with hydrazine hydrate to give a compound of Formula XXII can be carried out at a temperature ranging, for example, from 120 to 140°C.
The reaction of a compound of Formula XXII with a compound of Formula HC(ORn)3 to give a compound of Formula XXIII can be carried out at a temperature ranging, for example, from 60tol60°C
An illustrative list of compounds of the inyention is listed below
2-{3-[3-(Benzyloxy)-4-(difluoromethoxy)phenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no 1),
2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (compound no 2),
Ethyl {2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetate (compound no 3),
2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (compd no 4),
Ethyl {2-(difluoromethoxy)-5-[5-methyl-5-(l,3J4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy} acetate (compound no. 5),
2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethanol (compound no 6),
4-(2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethyl)morpholme (compound no 7),
2-{3-[3-(Benzyloxy)-4-methoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no. 8),
2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetamide (compound no 9),
2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethanol (compound no 10),
2-{(5Sor 5i?)-3-[4-(Difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no 11),
2-{(5R or5S )-3-[4-(Difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no 12),
pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers,
diastereomers or N-oxides thereof.
The following compounds can be prepared by following the schemes of the inyention:
4-(2- {2-(Difluoromethoxy)-5-[5-methyl-5 -(1,3,4-oxadiazol-2-yl)-4,5 -dihydroisoxazol-3 -yl]phenoxy}ethyl)morpholine (compound no 13),
2-{2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetamide (compound no 14),
pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers,
diastereomers or N-oxides thereof
Where desired, the compounds of Formula I and/ or their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides may be advantageously used in combination with one or more other therapeutic agents. Examples of other therapeutic agents, which may be used in combination with compounds of Formula I of this inyention and/ or their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides include one other active ingredients selected from corticosteroids, B2- agonist, leukotnene antagonists, 5-lipoxygenase inhibitors, chemokine inhibitors, muscarinic receptor antagonists, p38 MAP kinase inhibitors, anticholinergics, antiallergics, PAF antagonists, EGFR kinase inhibitors, additional PDE-IV inhibitors, kinase inhibitors or combinations thereof.
The one or more 152- agonist as described herein may be chosen from those described in the art. The 132-agonists may include one or more compounds described in U S Patent Nos 3,705,233,
3,644,353, 3,642,896, 3,700,681, 4,579,985; 3,994,974; 3,937,838; 4,419,364, 5,126,375, 5,243,076, 4,992,474, and 4,011,258
132-agonists include, for example, one or more of albuterol, salbutamol, biltolterol, pirbuterol, levosalbutamol, tulobuterol, terbutaline, bambuterol, metaproterenol, fenoterol, salmeterol, carmoterol, arformoterol, formoterol, and their pharmaceutically acceptable salts or solvates thereof.
Corticosteroids as described herein may be chosen from those described in the art Corticosteroids may be include one or more compounds described in U S. Patent Nos 3,312,590, 3,983,233, 3,929,768; 3,721,687; 3,436,389; 3,506,694; 3,639,434; 3,992,534; 3,928,326; 3,980,778, 3,780,177; 3,652,554, 3,947,478, 4,076,708; 4,124,707; 4,158,055; 4,298,604, 4,335,121, 4,081,541, 4,226,862, 4,290,962; 4,587,236; 4,472,392, 4,472,393; 4,242,334; 4,014,909; 4,098,803; 4,619,921; 5,482,934; 5,837,699; 5,889,015; 5,278,156; 5,015,746, 5,976,573; 6,337,324; 6,057,307; 6,723,713; 6,127,353, and 6,180,781. The disclosures of these patents are incorporated herein by reference in their entireties.
Corticosteroids may include, for example, one or more of alclometasone, amcinonide, amelometasone, beclometasone, betamethasone, budesomde, ciclesonide, clobetasol, cloticasone, cyclomethasone, deflazacort, deprodone, dexbudesonide, diflorasone, difluprednate, fluticasone, flunisolide, halometasone, halopredone, hydrocortisone, methylprednisolone, mometasone, prednicarbate, prednisolone, nmexolone, tixocortol, triamcinolone, tolterodine, oxybutynin, ulobetasol, rofleponide, GW 215864, KSR 592, ST-126, dexamethasone and pharmaceutically acceptable salts, solvates thereof. Preferred corticosteroids include, for example, flunisolide, beclomethasone, triamcinolone, budesomde, fluticasone, mometasone, ciclesonide, and dexamethasone Examples of possible salts or derivatives include: sodium salts, sulfobenzoates, phosphates, isonicotinates, acetates, propionates, dihydrogen phosphates, palmitates, pivalates, or furoates In some cases, the corticosteroids may also occur in the form of their hydrates
Muscarinic receptor antagonists include substances that directly or indirectly block activation of muscarinic cholinergic receptors Examples include, but are not limited to, quaternary amines (eg, methantheline, ipratropium, propantheline), tertiary amines (eg, dicyclomine, scopolamine) and tricyclic amines (eg, telenzepine) Other suitable muscarinic receptor antagonists include benztropine (commercially available as COGENTIN from Merck), hexahydro-sila-difemdol hydrochloride (HHSID hydrochloride disclosed in Lambrecht et al, Trends in Pharmacol Sci, lO(Suppl) 60 (1989), (+/-)-3-quinuchdinyl xanthene-9-carboxylate hemioxalate (QNX-hemioxalate, Birdsall et al, Trends in Pharmacol Sci, 4 459 (1983); telenzepine dihydrochlonde (Coruzzi et al, Arch Int Pharmacodyn Ther, 302-232 (1989), and Kawashima et al, Gen Pharmacol ,2117 (1990)), and atropine
Anticholinergics include, for example, tiotropium salts, ipratropium salts, oxitropium salts, salts of the compounds known from WO 02/32899. tropenol N-methyl-2,2-diphenylpropionate, scopine N-methyl-2,2-diphenylpropionate, scopine N-methyl-2-fluoro-2,2-diphenylacetate and tropenol N-methyl-2-fluoro-2,2-diphenylacetate; as well as salts of the compounds known from WO 02/32898- tropenol N-methyl-3,3',4,4'-tetrafluorobenzilate, scopine N-methyl-3,3',4,4'-tetrafluorobenzilate, scopine N-methyl-4,4'-dichlorobenzilate, scopine N-methyl-4,4'-difluorobenzilate, tropenol N-methyl-3,3'-difluorobenzilate, scopine N-methyl-3,3'-difluorobenzilate, and tropenol N-ethyl-4,4'-difluorobenzilate, optionally in the form of their hydrates and solvates By salts are meant those compounds which contain, in addition to the above mentioned cations, as counter-ion, an anion with a single negative charge selected from among the chloride, bromide, and methanesulfonate.
Antiallergic agents include, for example, epinastine, cetinzine, azelastine, fexofenadine, levocabastine, loratadine, mizolastine, ketotifene, emedastme, dimetindene, clemastine, bamipine, hexachlorophemramine, pheniramine, doxylamine, chlorophenoxamine, dimenhydnnate, diphenhydramine, promethazine, ebastine, desloratadine, and meclizine Preferred antiallergic agents include, for example, epinastine, cetirizine, azelastine, fexofenadine, levocabastine, loratadine, ebastine, desloratadine, and mizolastine. Any reference to the above-mentioned antiallergic agents also includes any pharmacologically acceptable acid addition salts thereof, which may exist
PAF antagonists include, for example, 4-(2-chlorophenyl)-9-methyl-2-[3-(4-morphohnyl)-3-propanon-l-yl]-6H-thieno[3,2-fJ[l,2,4]triazolo[4,3-a][l,4]diazepine and 6-(2-chlorophenyl)-8,9-dihydro-l-methyl-8-[(4-morpholinyl)carbonyl]-4H,7H-cyclopenta[4.5]thieno[3,2-fj[l,2,4]tnazolo[4,3-a][l,4]diazepine
EGFR kinase inhibitors include, for example, 4-[(3-chloro-4-fluorophenyl)amino]-7-(2-{4-[(S)-(2-oxotetrahydrofuran-5-yl)carbonyl]piperazin-l-yl}-ethoxy)-6-
[(vinylcarbonyl)amino]quinazoline, 4-[(3-chloro4-fluorophenyl)amino]-7-[4-((S)-6-methyl-2-
oxomorphohn-4-yl)butyloxy] -6- [(vinylcarbonyl)amino] quinazohne, 4- [(3 -chloro4-
fiuorophenyl)amino]-7-[4-((R)-6-methyl-2-oxomorpholin-4-yl)butyloxy]-6-
[(vinylcarbonyl)amino]quinazoline, 4-[(3-chloro-4-fluorophenyl)amino]-7-[2-((S)-6-methyl-2-
oxomorpholm-4-yl)ethoxy]-6-[(vinylcarbonyl)amino]quinazolme, 4-[(3-chloro-4-
fluorophenyl)ammo]-6-[(4-{N-[2-(ethoxycarbonyl)ethyl]-N-[(ethoxycarbonyl)methyl]-amino}-1-oxo-2-buten-1 -yl)amino] -7-cyclopropylmethoxy quinazohne, 4- [(R)-( 1 -phenyl ethyl)amino] -6- {[4-(morphohn-4-yl)-1 -oxo-2-buten-1 -yljamino}-7-cyclopropyl-methoxyquinazoline, and 4-[(3-chloro-4-fluorophenyl)amino]-6-[3-(morphohn-4-yl)propyloxy]-7-methoxyquinazoline. Any reference to the above-mentioned EGFR kinase inhibitors also includes any pharmacologically acceptable acid
addition salts thereof which may exist By the physiologically or pharmacologically acceptable acid addition salts thereof which may be formed by the EGFR kinase inhibitors are meant, according to the inyention, pharmaceutically acceptable salts selected from among the salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfomc acid, acetic acid, fumanc acid, succinic acid, lactic acid, citric acid, tartaric acid, or maleic acid The salts of the EGFR kinase inhibitors selected from among the salts of acetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and methanesulfonic acid are preferred according to the inyention
p38 kinase inhibitors include, for example, l-[5-tert-butyl-2-p-tolyl-2H-pyrazol-3-yl]-3-[4-
(2-morpholin-4-ylethoxy)naphthalen-1 -yl]urea; 1 -[5-tert-butyl-2-p-tolyl-2H-pyrazol-3-yl]-3-[4-(2-
(1 -oxothiomorphohn-4-yl)ethoxy)naphthalen-1 -yl]urea; 1 -[5-tert-butyl-2-(2-methylpyndin-5-yl)-
2H-pyrazol-3-yl]-3-[4-(2-pyridin-4-ylethoxy)naphthalen-l-yl]urea; l-[5-tert-butyl-2-(2-
methoxypyridin-5-yl)-2H-pyrazol-3-yl]-3-[4-(2-morphohn-4-ylethoxy)naphthalen-1 -yl]urea, and 1 -
[5-tert-butyl-2-methyl-2H-pyrazol-3-yl]-3-[4-(2-morpholin-4-ylethoxy)naphthalen-l-yl]urea
disclosed in our co-pending US patent application no 60/605,344; 4-[7-Oxo-8-(tetrahydro-pyran-4-
yl)-6-o-tolyl-7,8-dihydro-pyrido[2,3-d]pyrimidin-2-ylammo]-piperidine-l -carboxyhc acid tert-
butyl ester; Hydrochloride salt of 2-(Pipendin-4-ylamino)-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-8H-
pyrido[2,3-d]pynmidin-7-one, 2-(l-Methanesulfonyl-pipendin-4-ylamino)-8-(tetrahydro-pyran-4-
yl)-6-o-tolyl-8H-pyndo[2,3-d]pynmidin-7-one, 2-(l-Benzyl-piperidin-4-ylamino)-8-(tetrahydro-
pyran-4-yl)-6-o-tolyl-8H-pyrido[2,3-d]pyrimidin-7-one; 2-(l-Methyl-pipendin-4-ylamino)-8-
(tetrahydro-pyran-4-yl)-6-o-tolyl-8H-pyrido[2,3-d]pyrimidin-7-one, 2-(4-Methyl-piperazin-l-
ylamino)-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-8H-pyrido[2,3-d]pyrimidin-7-one; 4-[6-(2-Chloro-
phenyl)-7-oxo-8-(tetrahydro-pyran-4-yl)-7,8-dihydro-pyndo[2,3-d]pyrimidin-2-ylamino]-
pipendine-1 -carboxyhc acid tert-butyl ester, 2-(Pipendin-l-ylamino)-8-(tetrahydro-pyran-4-yl)-6-
o-tolyl-8H-pyndo[2,3-d]pyrimidin-7-one; 2-Cyclobutylamino-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-
8H-pyrido[2,3-d]pyrimidin-7-one; 2-(l-Acetyl-piperidin-4-ylamino)-8-(tetrahydro-pyran-4-yl)-6-o-
tolyl-8H-pyndo[2,3-d]pyrimidin-7-one; 2-(l-Benzoyl-piperidin-4-yiamino)-8-(tetrahydro-pyran-4-
yl)-6-o-tolyl-8H-pyrido[2,3-d]pynmidin-7-one, 2-(l-Benzoyl-piperidin-4-ylamino)-8-(tetrahydro-
pyran-4-yl)-6-o-tolyl-8H-pyndo[2,3-d]pynmidin-7-one; 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-
tolyl-7,8-dihydro-pyrido[2,3-d]pyrimidin-2-ylamino]-pipendine-1 -carboxyhc acid (4-fluoro-
phenyl)-amide; 2-( 1 -Ethanesulfonyl-pipendin-4-ylamino)-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-8H-
pyndo[2,3-d]pynmidin-7-one, 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-
pyndo[2,3-d]pynmidin-2-ylamino]-piperidine-l-carbothioic acid (4-fluoro-phenyl)-amide; 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyrido[2,3-d]pyrimidin-2-ylamino]-pipendine-1-carboxyhc acid (4-trifluoromethyl-phenyl)-amide, 2-[4-(Propane-2-sulfonyl)-
piperazin-l-ylamino]-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-8H-pyrido[2,3-d]pynmidin-7-one; 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyrido[2,3-d]pyrimidin-2-ylamino]-piperazme-1-carboxylic acid propylamide, 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyndo[2,3-d]pynmidin-2-ylamino]-piperazine-l-carboxylic acid ((R)-l ,2-dimethyl-propyl)-amide, 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyrido[2,3-d]pyrimidin-2-ylammo]-piperazine-l-carboxylic acid cyclohexylamide; 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyndo [2,3 -d]pyrimidin-2-ylamino] -piperazine-1 -carboxyhc acid (4-fluoro-phenyl)-amide; 4-[7-Oxo-8-(tetrahydro-pyran-4-yl)-6-o-tolyl-7,8-dihydro-pyrido[2,3-d]pynmidin-2-ylamino]-piperazine-l-carboxyhc acid cyclopentyl methyl-amide; and the compounds which are disclosed m our co-pending US patent application no. 60/598621, 60/630,517 and Indian patent application no 1098/DEL/2005 and 211/DEL/2005. Any reference to the above mentioned p38 kinase inhibitors also includes any pharmacologically acceptable acid addition salts thereof which may exist By the physiologically or pharmacologically acceptable acid addition salts thereof which may be formed by the p38 kinase inhibitors are meant, according to the inyention, pharmaceutically acceptable salts selected from among the salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, acetic acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, and maleic acid.
Additional PDE-IV inhibitors include, for example, enprofylhne, roflumilast, oglemilast, ariflo, Bay-198004, CP-325,366, BY343, D-4396 (Sch-351591), V-11294A, Z-15370, and AWD-12-281 Preferred PDE-IV inhibitors are selected from among enprofylline, roflumilast, ariflo, Z15370, and AWD-12-281. Any reference to the above mentioned PDE-IV inhibitors also includes any pharmacologically acceptable acid addition salts thereof which may exist. By the physiologically acceptable acid addition salts which may be formed by the above mentioned PDE-IV inhibitors are meant, according to the inyention, pharmaceutically acceptable salts selected from among the salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, acetic acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, or maleic acid According to the inyention, the salts selected from among the acetate, hydrochloride, hydrobromide, sulfate, phosphate, and methanesulfonate are preferred in this context
The leukotnene antagonist can be selected from compounds not limited to those described mUS 5,565,473, US 5,583,152, US 4,859,692 or US 4,780,469
Examples of leukotnene antagonist include, but are not limited to, montelukast, zafirlukast, pranlukast and pharmaceutically acceptable salts thereof.
5-Lipoxygenase inhibitors can be selected from the compounds disclosed in U S. 4,826,868, 4,873,259, EP 419049, EP 542356 or EP 542355. Examples may include but are not limited to atreleuton, zyflo (zileuton), ABT-761, fenleuton or tepoxalin
Chemokine inhibitors can be selected from the compounds disclosed in EP 287436, EP 389359, EP 988292, WO 02/26723 or WO 01/90106.
Examples of chemokine inhibitors include, but are not limited to AMD3100, AZD 8309, BX-471, GW-766994, UK-427857, CP-481715, UK-107543, UK-382055 or UK-395859.
Examples set forth below demonstrate the synthetic procedures for the preparation of the representative compounds. The examples are provided to illustrate particular aspect of the disclosure and do not constrain the scope of the present inyention as defined by the claims.
Experimental details Example 1 Preparation of 3 -hydroxy-4-difluoromethoxybenzaldehyde
Benzyltriethyl ammonium chloride (4.12 g, 0.0182 mol) was added to a solution of 3,4-dihydroxy benzaldehyde (5g, 0.0362 mol) in dimethylformamide (35 mL) Sodium hydroxide solution (0.0905 mol of 30% solution) was added dropwise to the resulting reaction mixture for about 10 minutes with a continuous flow of chloro-difluoro methane The reaction mixture was acidified with dilute hydrochloric acid and then diluted with water It was extracted with ethyl acetate, washed with saturated solution of sodium chloride and concentrated under reduced pressure The residue thus obtained was purified by column chromatography using 10% ethyl acetate in hexane to furnish the title compound Y1eld 2.5 g (37%) Example 2 Preparation of 3-benzyloxy-4-methoxybenzaldehyde
The title compound was prepared according to the method described in J. Med Chem 1994,37,1696-1703
The following compound was prepared by following above procedure: 3-(Benzyloxy)-4-(difluoromethoxy)benzaldehyde Y1eld 99% Example 3- Preparation of 3-benzyloxy-4-methoxybenzaldehyde oxime
Hydroxylamine hydrochloride (50 25 g, 0.723 mol) and sodium acetate (59 31 g, 0 723 mmol) were added to a stirred solution of 3-benzyloxy-4-methoxybenzaldehyde (35 g, 0 144 mol) (example 2) in ethanol (200 mL). The reaction mixture was allowed to stir at room temperature for about 50 minutes Ethanol was removed under reduced pressure, residue was poured in water (250 mL) and extraction was done with ethyl acetate (2 x 150 mL). Ethyl acetate layer was dried over anhydrous sodium sulphate, filtered and finally concentrated under reduced pressure to afford title compound Y1eld: 36 g (96.8%) The following compound was prepared by following above procedure:
3 -(Benzyloxy)-4-(difluoromethoxy)benzaldehyde oxime
Y1eld- 99%
Example 4: Preparation of methyl 3-[3-(benzyloxy)-4-methoxyphenyl"|-5-methyl-4,5-
dihydroisoxazole-5-carboxylate
3-Benzyloxy-4-methoxybenzaldehyde oxime (10 g, 0 0389 mol) (example 3) was taken in tetrahydrofuran (50 mL) Methyl methacrylate (8.3 mL, 0.0778 mol) was added at room temperature. Sodium hypochlorite solution (100 mL) was added dropwise. The reaction mixture was stirred vigorously for about 14 hours at an ambient temperature. Tetrahydrofuran was removed under reduced pressure Water was added and extraction was done with ethyl acetate. The organic layer was dried and concentrated in vacuo. The residue was purified by column chromatography using ethyl acetate and hexane (30 70) Y1eld- 12 5 g (93 6%)
The following compounds were prepared by following above procedure Methyl 3-[3-(benzyloxy)-4-(difluoromethoxy)phenyl]-5-methyl-4,5-dihydroisoxazole-5-carboxylate Y1eld 81%
Methyl 3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carboxylate Y1eld 83 2%
Example 5: Preparation of 3-[3-(benzyloxy)-4-methoxyphenyl1-5-methyl-4,5-dihydroisoxazole-5-carbohydrazide
Hydrazine-hydrate (10 mL) was added to methyl 3-[3-(benzyloxy)-4-methoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carboxylate (1 0 g, 0.0029 mol) (example 4). The reaction mixture was heated overnight at about 120°C. It was cooled, water was added and extraction was done with ethyl acetate The organic layer was dried and concentrated in vacuo Y1eld • 800 mg (77%)
The following compound was prepared by following above procedure:
3-[3-(Benzyloxy)-4-(difluoromethoxy)phenyl]-5-methyl-4,5-dihydroisoxazole-5-carbohydrazide Y1eld 89%
Example 6- Preparation of 2-{3-[3-(benzyloxy)-4-methoxyphenyl1-5-methyl-4,5-dihydroisoxazol-5-yl|-L3.4-oxadiazole (compound no 8)
Triethylorthoformate (5 mL) was added to 3-[3-(benzyloxy)-4-methoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carbohydrazide (200 mg) (example 5) The reaction mixture was heated at about 120°C for about 3 hours Excess triethylorthoformate was evaporated and the residue was heated at about 140°C for about 2 hours. The reaction mixture was diluted with water, saturated
with potassium carbonate and extracted with ethyl acetate. The organic layer was dried,
concentrated and purified by column chromatography (ethyl acetate • hexane- 70-30).
Y1eld- 150mg(73%).
1HNMR (CDCI3). 2 04 (s, 3H), 3.39-3 44 (d, 1H), 3.92 (s, 3H), 4.16-4.21 (d, 1H), 5.16 (s, 3H),
6.88 (d, 1H, ArH), 7.09-7.12 (m, 1H, ArH), 7 26-7.46 (m, 6H, ArH), 8.43 (s, 1H).
The following compound was prepared by following the above procedure.
2- {3 - [3 -(Benzyloxy)-4-(difluoromethoxy)phenyl]-5 -methyl-4,5-dihydroisoxazol-5-yl} -1,3,4-
oxadiazole (compound no 1)
Y1eld- 57%
Example 7- Preparation of 2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yiy4,5-dihydroisoxazol-3-
yl] phenol (compound no 4)
Palladium/carbon (200 mg) was added to a solution of 2-[3-(3-benzyloxy-4-memoxyphenyl)-5-methyl-4,5-dihydroisoxazol-5-yl]-[l,3,4]oxadiazole (150 mg) (example 6) in methanol (5 mL). Hydrogen gas was perged through balloon The reaction mixture was stirred in hydrogen atmosphere for about 3-4 hrs at room temperature The catalyst palladium/carbon was filtered through celite and the mixture was washed with methanol. The organic solvent was concentrated under vacuo to give title compound Y1eld. 30mg(29%).
1HNMR. (CDCI3) 1 97 (s, 3H), 3 42-3 46 (d, 1H), 3.93 (s, 3H), 4 18-4.22 (d, 1H), 6.87 (d, 1H), 7 17-7 20 (m, 1H), 7 29 (s, 1H), 8.44 (s, 1H) M++l 276 2
The following compound was prepared by following above procedure: 2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (compound no 2) Y1eld 20%
1HNMR (MeOD) 1 931 (s, 3H), 3 59-3 63 (d, 1H), 4.10-4 15 (d, 1H), 6.65-7.02 (t, 1H), 7.13-7.18 (m, 2H), 7.3 (s, 1H), 8 99 (s, 1H). M++l 312.1
Example 8- Preparation of ethyl {2-methoxy-5-[5-methyl-5-('l,3,4-oxadiazol-2-vlV4.5-dihydroisoxazol-3-yllphenoxylacetate (compound no 3)
Potassium carbonate (100 mg, 0 00072 mol) and bromoethyl acetate (0 05 mL, 0 00043 mol) were added to 2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (80 mg, 0.00029 mol) (example 7) in dimethylformamide (1 mL) The reaction mixture was stirred overnight at room temperature Water was added and the extraction was done
with ethyl acetate The organic layer was dried over anhydrous sodium sulphate, concentrated in
vacuo and the residue was purified by column chromatography (ethyl acetate : hexane :• 50:50).
Y1eld 50 mg (48%)
1HNMR: (CDC13) 1 24-1 31 (t, 3H), 1 97 (s, 3H), 3 42-3 46 (d, 1H), 4 09 (s, 3H), 4 11-4.15 (d,
1H), 4 22-4 29 (q, 2H), 4.71 (s, 2H), 6.91 (d, 1H), 7.15 (d, H), 7.30 (s, 1H), 8 44 (s, 1H).
M++1.362 1
The following compounds were prepared by following above procedure.
Ethyl {2-(difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-
yl]phenoxy} acetate (compound no 5)
1HNMR: (CDCI3) 1.26-1 32 (t, 3H), 1 98 (s, 3H), 3.45 (d, 1H), 4.20-4.29 (m, 3H), 4.75 (s, 1H),
6.58-6 96 (t, 1H), 7 14 (d, 1H), 7.23-7 27 (m, 1H), 7 38 (s, 1H), 8.46 (s, 1H).
M++l:397 87
2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethanol
(compound no 6)
1HNMR (CD3OD): 1 91 (s, 3H), 3.58 (d, 1H), 3.87 (d, 1H), 3 91 (s, 3H), 4 07-4.17 (m, 4H), 6 97
(d, 1H), 7 24 (d, 1H), 7 37 (s, 1H), 8 97 (s, 1H).
M++1:320 0
2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-
yl]phenoxy}ethanol (compound no. 10)
1HNMR (CDCI3): 1 98 (s, 3H), 2.91 (d, 1H), 3.45 (d, 1H), 3 98-4.01 (t, 2H), 4 16-4.19 (t, 2H),
6 36-6.85 (t, 1H), 7 11-7 14 (m, 1H), 7 20-7 26 (m, 1H), 7 47 (s, 1H), 8.45 (s, 1H)
M++l:355 98
4-(2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-
yl]phenoxy}ethyl)morphohne (compound no 7)
1HNMR (CDC13)- 1 98 (s, 3H), 2 64 (bs, 4H), 2 89-2.91 (m, 2H), 3.44-3.48 (d, 1H), 3 74-3.77 (m,
4H), 3 90 (s, 3H), 4 19-4.25 (m, 3H), 6.87-6.89 (d, 1H), 7.10 (d, 1H), 7 39 (s, 1H), 8.45 (s, 1H).
M++l 389 05
The following compound can be prepared by following above procedure:
4-(2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-
yl]phenoxy}ethyl)morpholine (compound no 13).
Example 9 Preparation of 2-{2-(difluQromethoxyV5-[5-methyl-5-(T,3,4-oxadiazol-2-yl)-4,5-
dihydroisoxazol-3-yl]phenoxy}acetamide (compound no 9)
Methanohc ammonia (5 mL) was added to ethyl {2-(difiuoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetate (30 mg) (example 8). The
reaction mixture was stirred overnight at room temperature. Methanol was evaporated off and the
residue was purified by preparative thin layer chromatography using ethyl acetate.
Y1eld 15mg(54%)
1HNMR. (CDCI3) 1.99 (s, 3H), 3.45 (d, 1H), 4.25 (d, 1H), 4.57 (s, 2H), 5.86 (bs, 1H), 6 31-6 80 (t,
1H), 7.21-7 26 (m, 3H), 8.45 (s, 1H)
M++l 368 98
The following compound can be prepared by following above method*
2-{2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetamide
(compound no. 14)
Example 10' Preparation of 3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-
dihydroisoxazole-5-carboxylic acid
Methyl 3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-
carboxylate ( 1 g, 0 0030 mol) (example 4) was taken in tetrahydrofuran (10 mL). Lithium hydroxide solution (382 mg, 0 0091 mol in 1 mL water) was added and the reaction mixture was stirred at room temperature overnight Tetrahydrofuran was removed under reduced pressure Water was added and the mixture was extracted with ethyl acetate. Aqueous layer was acidified by adding concentrated hydrochloric acid and extracted with ethyl acetate The organic layer was dried over anhydrous sodium sulphate and concentrated in vacuo to give title compound Y1eld- lg (crude)
Example 11 • Preparation of (5R or 55)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-A^-[(li?)-l-phenylethyl1-4,5-dihydroisoxazole-5-carboxamide
3-[4-(Difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carboxyhc acid (500 mg, 0.00158 mol) (example 10) and oxalyl chloride (0.413 mL, 0 00474 mol) in benzene (15 mL) were refluxed for about half an hour Benzene was evaporated off and the residue was diluted with dichloromethane (10 mL) The solution obtained was added to the solution of (s) (-) alpha methyl benzyl amine (0.425 mL, 0.00316 mol) in dichloromethane (10 mL) dropwise at about 0°C. The reaction mixture was stirred for about 1 hr at room temperature. The organic layer was washed with IN hydrochloric acid and then with 10 % sodium bicarbonate. It was extracted with water, dried and concentrated in vacuo to give mixture of title diastereomers (I and II) Diastereomers were then separated by column chromatography Y1eld (total) 67.8%
1HNMR diastereomer I (CDC13) 1 44 (m, 6H), 1 68 (s, 3H), 3 20 (d, 1H), 4.00 (d, 1H), 4 12-4.17 (q, 2H), 5 03-5.08 (m, 1H), 6 37-6 87 (t, 1H), 7 06-7 38 (m, 8H). Mass 419 02(M++1)
1HNMR: diastereomer II (CDC13) 1 42-1.52 (m, 6H), 1.74 (s, 3H), 3.19 (d, 1H), 3 74 (d, 1H),
4.07-4.14 (q, 2H), 5 05 (m, 1H), 6.35-6 85 (t, 1H), 7 01-7 32 (m, 8H)
Mass-419 02(M++1)
Example 12- Preparation of (5R or 5S)-3-(3-ethoxy-4-hydroxyphenyl)-5-methyl-4,5-
dihydroisoxazole-5-carboxylic acid
(5R or 55)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-N-[(1R)-l-phenylethyl]-4,5-dihydroisoxazole-5-carboxamide (500 mg, 0.00119 mol) (example 11) was taken in a mixture of isopropanol methanol (15 6mL). Hydrazine hydrate (8 9 mL, 0.1794 mol) and potassium hydroxide (10 g, 0.1794 mol) were added to the mixture. The reaction mixture was heated at refluxing temperature for about 48 hrs The organic solvent was removed under vacuo, water was added to the residue and it was acidified with concentrated hydrochloric acid The extraction was done with ethyl acetate (2 x 50 mL) and the mixture was basified with saturated sodium bicarbonate solution and again extracted with ethyl acetate The aqueous layer was acidified and extracted with ethyl acetate, dried and concentrated in vacuo to give respective enantiomers (enantiomer I and enantiomer II) Enantiomerl Y1eld 200mg(63%)
1HNMR- (CD3OD) 1.40-1 45 (t, 3H), 1.64 (s, 1H), 3.28-3.34 (m, 1H), 3.76 (d, 1H), 4.07-4.14 (q, 2H), 6.82 (d, 1H), 7 03-7 07 (m, 1H), 7.27 (s, 1H). Mass 266 08(M++1) Enantiomer II.
1HNMR (MeOD + D20) 1 41-1.46 (t, 3H), 1.67 (s, 1H), 3 40 (d, 1H), 3 83 (d, 1H), 4 09-4.16 (q, 2H), 6 88 (d, 1H), 7 07-7.10 (m, 1H), 7.29 (s, 1H) Mass. 266 05 (M++l)
Example 13' Preparation of methyl (5R or 5,S)-3-(3-ethoxy-4-hydroxyphenyl)-5-methyl-4,5-dihydroisoxazole-5-carboxylate
A mixture of (5R or J5)-3-(3-ethoxy-4-hydroxyphenyl)-5-methyl-4,5-dihydroisoxazole-5-carboxyhc acid (140 mg, 0 00052 mol) (example 12) and concentrated sulfuric acid (0 1 mL) in methanol was refluxed at 60-70°C for about 3 hrs Methanol was evaporated off The reaction mixture was diluted with water, extracted with ethyl acetate and washed with 10% solution of sodium bicarbonate The organic layer was dried over anhydrous sodium sulphate and concentrated in vacuo to give the respective enantiomers (enantiomer III and enantiomer IV) Enanatiomer III Y1eld 115mg (78%).
1HNMR- (CDC13)- 1.43-1 48 (t, 3H), 1 71 (s, 1H), 3 19 (d, 1H), 3 80 (s, 3H), 3.84 (d, 1H), 4 11-4 18 (q, 2H), 5 90 (bs, 1H), 6 89-6 97 (m, 2H), 7.37 (s, 1H). Mass- 280 06 (M++l)
Enanatiomer IV: Y1eld' 78%
1HNMR. (CDCb): 1.44-1.47 (t, 3H), 1.71 (s, 1H), 3.19 (d, 1H), 3.80 (s, 3H), 3.84 (d, 1H), 4 12-
4.17 (q, 2H), 5 90 (s, 1H), 6 90-6 99 (m, 2H), 7.36 (s, 1H).
Mass 279 99(M++1)
Example 14 Preparation of methyl (5R or 5S)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-
4,5-dihydroisoxazole-5-carboxylate
Freon gas was purged through a mixture of methyl (5R or 55)-3-(3-ethoxy-4-hydroxyphenyl)-5-methyl-4,5-dihydroisoxazole-5-carboxylate (110 mg, 0 00039 mol) (example 13), potassium carbonate (245 mg, 0.00078 mol) and benzyltriethylammonium chloride (8.0 mg, 0.000039 mol) in dimethylformamide (5 mL) at about -10°C for about 3-4 minutes The reaction mixture was stirred overnight at room temperature. It was diluted with water and extracted with ethyl acetate The organic layer was washed with brine, dried over anhydrous sodium sulphate and concentrated in vacuo to give respective enantiomers (enantiomer V and enantiomer VI). Enantiomer V- Y1eld 90 mg (70%)
1HNMR- (CDCI3). 1 43-1 48 (t, 3H), 1.72 (s, 1H), 3.19 (d, 1H), 3.81 (s, 1H), 3 86 (d, 1H), 4.09-4.16 (q, 2H), 6 36-6.86 (t, 1H), 7 00-7 03 (m, 1H), 7 17 (d, 1H), 7.43 (s, 1H). Mass. 329 97(M++1) Enantiomer VI Y1eld- 70%
1HNMR (CDCI3). 1 41-1.48 (t, 3H), 1 72 (s, 1H), 3 19 (d, 1H), 3.81 (s, 3H), 3 89 (d, 1H), 4.09-4 16 (q, 2H), 6.36-6 86 (t, 1H), 7 00-7.03 (m, 1H), 7 17 (d, 1H), 7.43 (s, 1H). Mass-329 97(M++1)
Example 15 Preparation of (5R or 550-3-[4-(difluoromethoxy)-3-ethoxyphenyl1-5-methyl-4,5-dihydroisoxazole- 5 -carbohydrazide
Hydrazine-hydrate (2 mL) was added to methyl (5R or 5S)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carboxylate (80 mg, 0 000243 mol) (example 14) The reaction mixture was heated at about 120°C for about 6 hrs. It was cooled and water was added Extraction was done with ethyl acetate, the organic layer was dried and concentrated in vacuo to give respective enantiomers (enantiomer VII and enantiomer VIII) Enantiomer VII: Y1eld : 60 mg (60%) Enantiomer VIII Y1eld 75%
Example 16- Preparation of 2-{(5R or 55V3-[4-(difluoromethoxyV3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5 -yl} -1,3,4-oxadiazole
Triethylorthoformate (2 mL) was added to (5R or 55)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazole-5-carbohydrazide (60 mg, 0 000182 mol) (example 15) The reaction mixture was heated at about 120°C for about 3 hours Excess
tnethylorthoformate was evaporated and the residue was heated at about 140°C for about 2 hours
The mixture was diluted with water, saturated with potassium carbonate and extracted with ethyl
acetate. The organic layer was dried, concentrated and purified by column chromatography The
residue was purified on crystallization by using diisopropyl ether to give the respective enantiomers
(enantiomer IX and enantiomer X).
Enantiomer IX: Y1eld . 15 mg (25%).
1HNMR: (CDC13). 1 43-1.48 (t, 3H), 1.98 (s, 3H), 3.45 (d, 1H), 4.10-4 17 (q, 2H), 4 16 (d, 1H),
6 37-6.87 (t, 1H), 7.06-7.09 (m, 1H), 7.19 (d, 1H), 7.38 (s, 1H), 8 44 (s, 1H)
Chiral purity: 98 22%
Mass. 339.96 (M++l)
Enantiomer X: Y1eld • 15%
1HNMR: (CDCI3): 1 44-1.48 (t, 3H), 1 98 (s, 3H), 3.45 (d, 1H), 4.07-4 14 (q, 2H), 4.19 (d, 1H),
6 37-6.87 (t, 1H), 7 06-7 09 (m, 1H), 7.20 (d, 1H), 7.44 (s, 1H), 8 44 (s, 1H)
Chiral purity: 98.85%
Mass- 339 96 (M++l)
Example 17 Efficacy of compounds
(a) PDE-IV Enzyme Assay
The efficacy of compounds of PDE-IV inhibitors was determined by an enzyme assay using U937 cell cytosolic fraction (Biochem Biophys. Res. Comm, 197: 1126-1131, 1993). The enzyme reaction was carried out in the presence of c AMP (1 uM) at 30°C in the presence or absence of test compound for 45 -60 min. An aliquot of this reaction mixture was taken further for the ELISA assay and the protocol of the kit followed to determine level of cAMP in the sample. The concentration of the cAMP in the sample directly correlated with the degree of PDE-4 enzyme inhibition Results were expressed as percent control and the IC50 values of test compounds were found to be in the range from about 10 uM to about 0 1 nM concentration.
(b) Cell based Assay for TNF-a release
Method of isolation of Human Peripheral Blood Mononuclear Cells CPBMNC) Human whole blood was collected in vacutainer tubes containing heparin or EDTA as an anti coagulant The blood was diluted (1:1) in sterile phosphate buffered saline and 10 mL was carefully layered over 5 mL Ficoll Hypaque gradient (density 1.077 g/mL) in a 15 mL conical centrifuge tube. The sample was centnfuged at 3000 rpm for 25 minutes in a swing-out rotor at room temperature After centrifugation, interface of cells were collected, diluted at least 1:5 with PBS (phosphate buffered saline) and washed three times by centrifugation at 2500 rpm for 10 minutes at room temperature. The cells were resuspended in serum free RPMI 1640 medium at a concentration of 2 million cells/mL
LPS (lipopolysaccharide) stimulation of Human PBMNC
PBMN cells (0 1 mL, 2 milhon/mL) were co-incubated with 20 ul of compound (final
DMSO concentration of 0 2 %) for 10 min in a flat bottom 96 well microtiter plate. Compounds
were dissolved in DMSO initially and diluted in medium for a final concentration of 0 2 % DMSO.
LPS (1 µg/mL, final concentration) was then added at a volume of 10 ul per well After 30 min, 20
ul of fetal calf serum (final concentration of 10 %) was added to each well. Cultures were
incubated overnight at 37°C in an atmosphere of 5 % CO2 and 95 % air Supernatant were then
removed and tested by ELISA for TNF-cc release using a commercial kit (e.g BD Biosciences) For
whole blood, the plasma samples were diluted 1 '20 for ELISA. The level of TNF-a in treated
wells was compared with the vehicle treated D controls and inhibitory potency of compound was
expressed as IC50 values calculated by using Graph pad prism. IC50 values of test compounds were
found to be in the range from about 10 uM to about 100 nM concentration
Percent TNF-a drug treated
Percent inhibition =100 x 100
Percent TNF-a in vehicle treated
(c) In-vitro assay to evaluate efficacy of PDEIV inhibitors in combination with p38 MAP
Kinase inhibitors
Perform the assay as described in (b) above, with individual compounds and their
combinations tested at sub-optimal doses.
(d) In-vitro assay to evaluate efficacy of PDE IV inhibitors in combination with 02- agonists
Measurement of Intracellular cAMP Elevation in U937 Cells
Grow U937 cells (human promonocytic cell line) in endotoxin-free RPMI 1640 + HEPES
medium containing 10% (v/v) heat-inactivated foetal bovine serum and 1% (v/v) of an antibiotic
solution (5000 IU/mL penicillin, 5000 µg/mL streptomycin). Resuspend cells (0 25 x 106/200 ul) in
Krebs' buffer solution and incubate at 37°C for 15 min in the presence of test compounds or vehicle
(20µl) Initiate generation of cAMP by adding 50 ul of 10 uM prostaglandin (PGE2) Stop the
reaction after 15 min, by adding 1 N HC1 (50 ul) and place on ice for 30 min Centrifuge the sample
(450g, 3 mm), and measure levels of cAMP in the supernatant using cAMP enzyme-linked
immunosorbent assay kit (Assay Designs) Calculate percent inhibition by the following formula
and calculate IC50 value using Graph pad prism.

(Formula Removed)

(e) In-vitro functional assays to evaluate efficacy of PDEIV inhibitors in combination with
beta-agonists
Animals and anaesthesia
Procure Guinea Pig (400-600gm) and remove trachea under anesthesia (sodium pentobarbital, 300 mg/kg 1 p) and immediately keep it in ice-cold Krebs Henseleit buffer. Indomethacin (10µM) is present throughout the KH buffer to prevent the formation of bronchoactive prostanoids Trachea experiments:
Clean the tissue off adherent fascia and cut it into strips of equal size (with approx 4-5 tracheal rings in each strip). Remove the epithelium by careful rubbing, minimizing damage to the smooth muscle Open the trachea along the mid-dorsal surface with the smooth muscle band intact and make a series of transverse cuts from alternate sides so that they do not transect the preparation completely Tie opposite end of the cut rings with the help of a thread Mount the tissue in isolated tissue baths containing 10 mL Krebs Henseleit buffer maintained at 37°C and bubbled with carbogen, at a basal tension of 1 g Change the buffer 4-5 times for about an hour Equilibrate the tissue for 1 hr with l^M carbachol or IO^IM histamine for stabilization. Wash it for 30 minutes followed by a precontraction with histamine (lOuM) or carbachol (luM) Allow the developed tension to stabilize for 15-20 minutes followed by the cumulative addition of beta-agonists prior to incubation with suboptimal dose of PDE IV inhibitor Record the contractile response of tissues either on Powerlab data acquisition system or on Grass polygraph (Model 7). Express the relaxation as percentage of maximum carbachol response Express the data as mean ± S E. mean for n observations Calculate the EC50 as the concentration producing 50% of the maximum relaxation to l^iM carbachol Compare percent relaxation between the treated and control tissues using non-parametric unpaired t-test A p value of < 0 05 is considered to be statistically significant.
(f) In-vivo assay to evaluate efficacy of PDE IV inhibitors m combination with beta-agonists
Lipopolysacchande (LPS) induced airway hyperreactivity (AHR) and neutrophilia'
Drug treatment
Beta-agonist (lng/kg to 1 mg/kg) and PDE4 inhibitor (lng/kg to 1 mg/kg) can be instilled intratracheally under anesthesia either alone or in combination.
Method
Use male wistar rats weighing 200±20gm in the study Rats should have free access to food and water On the day of experiment, expose animals to lipopolysacchande (LPS, lOOug/mL) for 40 mm Expose one group of vehicle treated rats to phosphate buffered saline (PBS) for 40 mm Two hours after LPS/PBS exposure, place animals inside a whole body plethysmograph (Buxco Electronics, USA) and expose to PBS or increasing acetylcholine (1,6, 12, 24, 48 and 96 mg/mL)
aerosol until Penh values (index of airway resistance) of rats attain 2 times the value (PC-100) seen
with PBS alone Record the respiratory parameters online using Biosystem XA software, (Buxco
Electronics, USA) Express Penh, at any chosen dose of acetylcholine is, as percent of PBS
response and using a nonlinear regression analysis compute PC 100 (2 folds of PBS value) values
Calculate percent inhibition using the following formula
(Formula Removed)
Where,
PC100LPS = PC 100 in vehicle treated group challenged group with LPS
PC100TEST = PC 100 in group treated with a given dose of test compound
PC100PBS = PC 100 in vehicle treated group challenged with PBS
Sacrifice animals immediately after recording the airway hyperreactivity response and perform
bronchoalveolar lavage (BAL). Centrifuge the collected lavage fluid at 3000 rpm for 5 min, at 4°C
Collect pellet and resuspend in lmL HBSS Perform total leukocyte count in the resuspended
sample Use a portion of suspension for cytocentrifugation and staining with Leishmann's stain for
differential leukocyte count. Express total leukocyte and Neutrophil counts as cell count (millions
cells mL-1 of BAL) Compute percent inhibition using the following formula
(Formula Removed)
Where,
NCLPS = Percentage of neutrophil in vehicle treated group challenged with LPS
NCTEST =Percentage of neutrophil in group treated with a given dose of test compound
NCPBS = Percentage of neutrophil in vehicle treated group challenged with PBS
Compute ED50 from percent inhibition values using Graph Pad Prism software (Graphpad Software Inc ,USA). (g) In-vitro functional assay to evaluate efficacy of PDE-IV inhibitors in combination with
Muscarinic receptor antagonists
Animals and anaesthesia:
Procure Guinea Pig (400-600gm) and remove trachea under anesthesia (sodium pentobarbital, 300 mg/kg i p) and immediately keep in ice-cold Krebs Henseleit buffer. Indomethacm (10µM) is present throughout the KH buffer to prevent the formation of bronchoactive prostanoids
Trachea experiments
Clean the tissue off adherent fascia and cut it into strips of equal size (with approx 4-5 tracheal rings in each strip) Remove the epithelium by careful rubbing, minimizing damage to the
smooth muscle. Open the trachea along the mid-dorsal surface with the smooth muscle band intact and make a series of transverse cuts from alternate sides so that they do not transect the preparation completely Tie opposite end of the cut rings with the help of a thread Mount the tissue in isolated tissue baths containing 10 mL Krebs Henseleit buffer maintained at 37°C and bubbled with carbogen, at a basal tension of 1 g Change the buffer 4-5 times for about an hour. Equilibrate the tissue for 1 hr for stabilization After 1 hr, challenge the tissue with 1 uM carbachol Repeat this after every 2-3 washes till two similar consecutive responses are obtained At the end of stabilization, wash the tissues for 30 minutes followed by incubation with suboptimal dose of MRA/ Vehicle for 20 minutes prior to contraction of the tissues with luM carbachol and subsequently assess the relaxant activity of the PDE-IV inhibitor [10-9 M to 10-4 M] on the stabilized developed tension/response Record the contractile response of tissues either on Powerlab data acquisition system or on Grass polygraph (Model 7). Express the relaxation as percentage of maximum carbachol response. Express the data as mean ± S.E. for n observations. Calculate the EC50 as the concentration producing 50% of the maximum relaxation to 1 uM carbachol Compare percent relaxation between the treated and control tissues using non-parametric unpaired t-test A p value of < 0 05 is considered to be statistically significant (h) In-vivo assay to evaluate efficacy of PDE-IV inhibitors in combination with MRA inhibitors
Drug treatment
MRA (lng/kg to lmg/kg) and PDE-IV inhibitor (lng/kg to lmg/kg) can be instilled intratracheally under anesthesia either alone or in combination.
Method-
Use male wistar rats weighing 200±20gm in the study Rats should have free access to food
and water On the day of experiment, expose animals to lipopolysaccharide (LPS, lOOug/mL) for
40 min Expose one group of vehicle treated rats to phosphate buffered saline (PBS) for 40 min
Two hours after LPS/PBS exposure, place animals inside a whole body plethysmograph (Buxco
Electronics, USA) and expose to PBS or increasing acetylcholine (1, 6, 12, 24, 48 and 96 mg/mL)
aerosol until Penh values (index of airway resistance) of rats attain 2 times the value (PC-100) seen
with PBS alone Record the respiratory parameters online using Biosystem XA software, (Buxco
Electronics, USA) Express Penh, at any chosen dose of acetylcholine is, as percent of PBS
response and using a nonlinear regression analysis compute PC 100 (2 folds of PBS value) values
Calculate percent inhibition using the following formula
(Formula Removed)
Where,
PC l OOLPS = PC 100 in vehicle treated and LPS challenged group
PC100TEST = PC 100 in group treated with a given dose of test compound PC100PBS = PC 100 in vehicle treated group challenged with PBS
Sacrifice animals immediately after recording the airway hyperreactivity response and perform bronchoalveolar lavage (BAL) Centrifuge the collected lavage fluid at 3000 rpm for 5 min, at 4°C Collect pellet and resuspend in lmL HBSS. Perform total leukocyte count in the resuspended sample. Use a portion of suspension for cytocentrifugation and staining with Leishmann's stain for differential leukocyte count Express total leukocyte and Neutrophil counts as cell count (millions cells mL-1 of BAL) Compute percent inhibition using the following formula
(Formula Removed)
Where,
NCLPS = Percentage of neutrophil in vehicle treated group challenged with LPS NCTEST =Percentage of neutrophil in group treated with a given dose of test compound NCPBS = Percentage of neutrophil in vehicle treated group challenged with PBS
Compute ED50 from percent inhibition values using Graph Pad Prism software (Graphpad Software Inc.,USA).

WE CLAIM:

1. A compound having the structure of Formula I:

(Formula Removed)

their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers,
diastereomers or N-oxides, wherein
R1, R2 and R3 are independently selected from hydrogen or alkyl;
X1 and X2 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
aryl, aralkyl, heteroaryl, heterocyclyl, (heteroaryl)alkyl or (heterocyclyl)alkyl;
Y represents an oxygen atom, a sulphur atom, or NR (wherein R is selected from hydrogen,
alkyl, alkenyl, alkynyl, un(saturated) cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl,
(heteroaryl)alkyl, or (heterocyclyl)alkyl);
Y1 and Y2 are independently selected from hydrogen, alkyl, nitro, cyano, halogen, OR
(wherein R is the same as defined earlier), SR (wherein R is the same as defined earlier),
NHR (wherein R is the same as defined earlier), COOR' or COR' (wherein R' is hydrogen,
alkyl, alkenyl, alkynyl, (unsaturated cycloalkyl, aryl, aralkyl, heterocyclyl,
(heterocyclyl)alkyl, or (heteroaryl)alkyl);
Y1 and X2, X1 and Y2, Xi and X2 may together form a cyclic ring fused with the ring
containing 3-5 carbon atoms within the ring and having 1-3 heteroatoms selected from N, O
or S.
2. A compound, which is selected from:
- 2-{3-[3-(Benzyloxy)-4-(difluoromethoxy)phenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-
1,3,4-oxadiazole (compound no. 1),
2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (compound no. 2),
- Ethyl {2-methoxy-5-[5-methyl-5-(l ,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yljphenoxy}acetate (compound no. 3),
- 2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenol (compound no. 4),
- Ethyl {2-(difluoromethoxy)-5-[5-methyl-5-(l ,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yljphenoxy} acetate (compound no. 5),
- 2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethanol (compound no. 6),
- 4-(2-{2-Methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethyl)morpholine (compound no. 7),
- 2-{3-[3-(Benzyloxy)-4-methoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no. 8),
- 2- {2-(Difluoromethoxy)-5-[5-methyl-5-(l ,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetamide (compound no. 9),
- 2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l ,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethanol (compound no. 10),
- 2-{(5S or 5/?)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no. 11),
- 2-{(5i? or 5iS)-3-[4-(difluoromethoxy)-3-ethoxyphenyl]-5-methyl-4,5-dihydroisoxazol-5-yl}-l,3,4-oxadiazole (compound no. 12),
- 4-(2-{2-(Difluoromethoxy)-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}ethyl)morpholine (compound no. 13),
- 2-{2-methoxy-5-[5-methyl-5-(l,3,4-oxadiazol-2-yl)-4,5-dihydroisoxazol-3-yl]phenoxy}acetamide (compound no. 14),
and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates,
enantiomers, diastereomers or N-oxides.
3. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or 2, together with at least one pharmaceutically acceptable carrier, excipient or diluent.
4. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or 2 and at least one other active ingredient selected from corticosteroids, 132- agonist, leukotriene antagonists, 5-lipoxygenase inhibitors, chemokine inhibitors, muscarinic receptor antagonists, p38 MAP kinase inhibitors, anticholinergics, antiallergics, PAF antagonists, EGFR kinase inhibitors, additional PDE-IV inhibitors, kinase inhibitors or combinations thereof.
5. A compound of claim 1 or 2 for treating, preventing, inhibiting or suppressing an inflammatory condition or disease or CNS diseases, in a patient.
6. A pharmaceutical composition of claim 3 or 4 for treating, preventing, inhibiting or suppressing an inflammatory condition or disease or CNS diseases, in a patient.
7. A compound of claim 1 or 2 for the treatment, prevention, inhibition or suppression of CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease (COPD), psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome (ARDS), eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases in a patient.
8. A pharmaceutical composition of claim 3 or 4 for the treatment, prevention, inhibition or suppression of CNS diseases, AIDS, asthma, arthritis, bronchitis, chronic obstructive pulmonary disease (COPD), psoriasis, allergic rhinitis, shock, atopic dermatitis, Crohn's disease, adult respiratory distress syndrome (ARDS), eosinophilic granuloma, allergic conjunctivitis, osteoarthritis, ulcerative colitis and other inflammatory diseases in a patient.
9. A method for the preparation of a compound of Formula IX

(Formula Removed)

their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers,
diastereomers or N-oxides, the method comprising:
reacting a compound of Formula II with a compound of Formula X1 Z (wherein Z is
halogen) to give a compound of Formula III [wherein Xi (except hydrogen), Y1 and Y2 are
the same as defined in claim 1],
(Formula Removed)
reacting the compound of Formula III with a compound of Formula X2Z [wherein Z is halogen] to give a compound of Formula IV [wherein X2 (except hydrogen) is same as defined in claim 1],
(Formula Removed)

reacting the compound of Formula IV with hydroxylamine hydrochloride to give a compound of Formula V,

(Formula Removed)
treating the compound of Formula V with a compound of Formula VI to give a compound of Formula VII [wherein Rj and R2 are the same as defined in claim 1 and Rr represents COOH, COOCH3],
(Formula Removed)
reacting the compound of Formula VII (when Rr is COOCH3) with hydrazine hydrate to give a compound of Formula VIII,
(Formula Removed)
reacting the compound of Formula VIII with a compound of Formula HC(OR3)3 to give the compound of Formula IX [wherein R3 is the same as defined in claim 1], or debenzylating a compound of Formula X to give a compound of Formula XI [wherein X1, Y1, Y2, R1, R2 and R3 are the same as defined in claim 1],
(Formula Removed)
reacting the compound of Formula XI with X2Z [wherein Z is halogen] to give the compound of Formula IX [wherein X2 (except hydrogen and benzyl) is same as defined in claim 1].

10. A method for the preparation of a compound of Formula XIII
(Formula Removed)

their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides, the method comprising: amidating a compound of Formula XII
(Formula Removed)

to give the compound of Formula XIII [wherein X1, Y1, Y2, R1, R2 and R3 are the same as defined in claim 1].
11. A method for the preparation of compounds of Formula XVI and Formula XVII

(Formula Removed)
their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides, the method comprising: reacting a compound of Formula XIV with a compound of Formula XV
(Formula Removed)
Formula XIV to give the compound of Formula XVI [wherein X1, X2, Y1, Y2 and R1 are the same as defined in claim 1] and the compound of Formula XVII [wherein X1, X2, Y1, Y2 and R1 are the same as defined in claim 1].

12. A method for the preparation of a compound of Formula XXIII
(Formula Removed)
Formula XXIII their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, enantiomers, diastereomers or N-oxides, the method comprising:
reacting a compound of Formula XVIII [wherein configuration at stereogenic carbons marked * is (R) or (S)] with hydrazine hydrate to give a compound of Formula XIX,
(Formula Removed)
reacting the compound of Formula XIX with methanol to give a compound of Formula XX,
(Formula Removed)
reacting the compound of Formula XX with Freon gas to give a compound of Formula XXI,

(Formula Removed)
Formula XXI reacting the compound of Formula XXI with hydrazine hydrate to give a compound of Formula XXII,

(Formula Removed)
reacting the compound of Formula XXII with a compound of Formula HC(OR3)3 to give the compound of Formula XXIII [wherein X2, Y1, Y2, R1 and R3 are the same as defined in claim 1 and configuration at stereogenic carbon marked * is (R) or (S)].